WO2021262956A1

WO2021262956A1 - Topical gel formulations and their use in treating skin conditions

Info

Publication number: WO2021262956A1
Application number: PCT/US2021/038858
Authority: WO
Inventors: Martin Osterhout; Meera G. PATEL; James Lee; Adam Simpson
Original assignee: Roivant Sciences Gmbh
Priority date: 2020-06-24
Filing date: 2021-06-24
Publication date: 2021-12-30

Abstract

Embodiments herein are directed to topical compositions comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. The topical compositions may be used to treat a variety of skin conditions, including atopic dermatitis, vitiligo, and alopecia areata.

Description

TOPICAL GEL FORMULATIONS AND THEIR USE IN TREATING SKIN CONDITIONS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and benefit of U.S. Provisional Application No. 63/043,340 filed June 24, 2020 and of U.S. Provisional Application No. 63/043,360 filed June 24, 2020, the entire contents of which are hereby incorporated by reference.

SUMMARY

[0002] Embodiments herein are directed to topical compositions comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300.

[0003] Some embodiments herein are directed to methods of treating skin conditions in a patient in need thereof comprising topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In certain embodiments, the skin condition is atopic dermatitis.

DESCRIPTION OF THE DRAWINGS

[0004] FIG. 1 depicts the drug level of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid in homogenized skin 12 hours post dose.

[0005] FIG. 2 depicts the estimated free fraction of drug (methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid) concentration.

DETAILED DESCRIPTION

[0006] This invention is not limited to the particular processes, compositions, or methodologies described, as these may vary. The terminology used in the description is for the purpose of describing the particular versions or embodiments only, and is not intended to limit the scope of the present invention. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art. All publications mentioned herein are incorporated by reference in their entirety. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

[0007] It must be noted that, as used herein, and in the appended claims, the singular forms “a,” “an,” and “the” include plural reference unless the context clearly dictates otherwise.

[0008] As used herein, the term “about” means plus or minus 10% of the numerical value of the number with which it is being used. Therefore, about 50% means in the range of 45% to 55%.

[0009] “Administering” when used in conjunction with a composition means to administer a composition to a patient whereby it positively impacts the tissue to which it is targeted, e.g. the skin. “Administering” a composition may be accomplished by, for example, topical administration, or in combination with other known techniques. Administering may be self-administration, wherein the subject in need of such treatment administers a composition or administering may be by a medical or other health care professional or a caretaker of the subject in need of such treatment.

[0010] The term “animal,” “patient,” or “subject” as used herein includes, but is not limited to, humans and non-human vertebrates such as wild, domestic and farm animals.

[0011] The term “acne” includes acne vulgaris and cystic acne and is characterized by areas of skin with seborrhea, comedones (blackheads and whiteheads), papules (pinheads), nodules (large papules) and pimples. Acne vulgarisis a disorder resulting from the action of hormones on the skin's oil glands (sebaceous glands), wherein the sebaceous glands of the skin produce excess sebum, and become enlarged and/or infected when the pore opening becomes plugged with a comedo (a mixture of keratin and sebum). The symptoms of acne include plugged pores and outbreaks of inflamed lesions commonly called pimples.

[0012] The term “alopecia areata” refers to a condition in which hair is lost from some or all areas of the body, also known as spot baldness.

[0013] The term “BHT” refers to butylated hydroxytoluene or dibutylhydroxytoluene.

[0014] As used herein, the terms “comprising,” “comprise,” “comprises,” and “comprised” are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0015] As used herein, the term “consists of’ or “consisting of’ means that the composition or method includes only the elements, steps, or ingredients specifically recited in the particular embodiment or claim. [0016] As used herein, the term “consisting essentially of’ or “consists essentially of’ means that the composition or method includes only the specified materials or steps and those that do not materially affect the basic and novel characteristics of the claimed invention.

[0017] Specific embodiments disclosed herein may be further limited in the claims using “consisting of’ or “consisting essentially of’ language, rather than “comprising”. In other words, though embodiments described herein use the phrase “comprising” or “comprises,” any embodiment described herein can be replaced with “consisting of ’/“consists of’ or “consisting essentially of ’/“consists essentially of.”

[0018] The term “dermatitis” is used to refer to a group of skin conditions which result in inflammation of the skin and is characterized by itchiness, red skin and a rash. Included in this group are atopic dermatitis, contact dermatitis, allergic contact dermatitis, irritant contact dermatitis, stasis dermatitis, seborrheic dermatitis, chronic dermatitis, and eczema.

[0019] The term “therapeutically effective amount” refers to an amount of a composition, of the embodiments described herein, necessary or sufficient to achieve the desired effect. For example, in some embodiments, the desired effect may include, without limitation, medically therapeutic, cosmetically therapeutic and/or prophylactic treatment, as appropriate.

[0020] The terms “exfoliative keratolysis” or “keratolysis exfoliative” refer to a skin condition which is characterized by dry skin and superficial, air-filled blisters. These blisters can be peeled off very easily and will leave reddish, tender areas.

[0021] “Follicular hyperkeratinization” plays a key role in the pathogenesis of acne, cells of the follicle become cohesive and do not shed normally onto the skin's surface and results in a microcomedone.

[0022] The term “ichthyosis” refers to a genetic skin disorder characterized by dry, thickened, and scaly skin.

[0023] In each of the embodiments disclosed herein, the compositions and methods may be utilized with or on a subject in need of such treatment, which may also be referred to as “in need thereof.” As used herein, the phrase “in need thereof’ means that the subject has been identified as having a need for the particular method or treatment and that the treatment has been given to the subject for that particular purpose.

[0024] The terms “keratosis follicularis” or “Darier's disease” refer to a genetic disorder characterized by dark crusty patches on the skin, sometimes containing pus. [0025] The term “Labrafil M1944CS” refers to the compound described as oleoyl polyoxylglycerides, oleoyl polyoxyl-6 glycerides, or oleoyl macrogol-6 glycerides.

[0026] The term “lichen simplex chronicus” refers to a skin disorder characterized by chronic itching and scratching. The constant scratching causes thick, leathery, darkened, (lichenified) skin.

[0027] The term “lichen planus” refers to a disease characterized by itchy reddish- purple polygon-shaped skin lesions on the lower back, wrists, and ankles.

[0028] As used herein, the term “methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid,” Έ6005,” or “RVT-501” shall also refer to alternative names of the compound, including N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic methyl ester, methyl 4-[(3-[6,7- dimethoxy-2-(methylamino)quinazolin-4-yl]phenyl)carbamoyl]benzoate, and methyl 4-[({3- [6,7-dimethoxy-2-(methylamino)quinazolin-4-yl]phenyl}amino)carbonyl]benzoate. The compound represented as RVT-501 or E6005 is methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid having the structure:

[0029] As used herein, the terms “metabolite of E6005,” “ER-392710,” or “Mil” refer to the metabolite of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid. The compound of Mil is 4-((3-(6,7-dimethoxy-2- (methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid and has the structure:

[0030] As used herein, the term “pharmaceutically acceptable” and grammatical variations thereof, as they refer to carriers, diluents, excipients, and reagents or other ingredients of the composition, represent that the materials used in the final composition are not irritating or otherwise harmful to the patient in general and to the skin, in particular, and preferably are pleasant and well tolerated with respect to general appearance, pH, color, smell and texture (feel), that they are not, for example, unacceptably sticky (tacky), oily or drying, and that they do spread easily, absorb into the skin at an acceptable rate of absorption.

[0031] The term “pityriasis rubra pilaris” refers to a group of chronic disorders characterized by reddish orange, scaling plaques and keratotic follicular papules. Symptoms may include reddish-orange patches on the skin, severe flaking, uncomfortable itching, thickening of the skin on the feet and hands, and thickened bumps around hair follicles.

[0032] The term “psoriasis” refers to the autoimmune disease characterized by patches of abnormal skin which is red, itchy and scaly. There are five main types of psoriasis: plaque, guttate, inverse, pustular, and erythrodermic.

[0033] The terms “pruritus” or “prurigo” refer to the severe itching of the skin due to a variety of ailments.

[0034] The term “palmoplantar pustulosis” refers to a chronic pustular condition affecting the palms and soles.

[0035] The term “rosacea” refers to a skin condition characterized by redness, pimples, swelling, and small, superficial dilated blood vessels.

[0036] The term “sebaceous adenomas” refers to a small bump on the skin, when many small bumps appear it is referred to as “sebaceous hyperplasia.”

[0037] The term “sebaceous gland” includes unilobular or multilobular glands that secrete sebum. Sebaceous glands include pilosebaceous units, fordyce spots, Meibomian glands, glands of the Zeiss and Montgomery areolar tubercles.

[0038] The phrase “disorder associated with sebaceous glands” includes diseases, conditions and symptoms related to sebaceous gland. Disorders associated with sebaceous glands include acne, seborrhea, sebaceoma, sebaceous carcinoma, seborrheic dermatitis, sebaceous cysts, sebaceous adenoma and sebaceous hyperplasia.

[0039] The term “seborrhea” includes oily skin.

[0040] The term “seborrheic dermatitis” includes inflammatory skin disorders characterized by scaly, flaky, itchy, and red skin and includes seborrheic dermatitis caused by fungal, genetic, environmental, hormonal and immune function disorders. [0041] The term “sebaceous cysts” include steatocystoma simplex (e.g., simple sebaceous duct cyst and solitary steatocystoma) and steatocystoma multiplex (e.g., epidermal polycystic disease and sebocystomatosis).

[0042] The term “sebaceous hyperplasia” includes enlargement of the sebaceous glands.

[0043] The term “skin” as used herein refers to the organ of the body which protects the subject from environmental irritations, regulates the body’s temperature and allows for external sensations. The “skin” is separated into three layers: the outermost layer called the epidermis which contains melanocytes; the dermis which contains connective tissue, hair follicles and sweat glands; and the deepest subcutaneous layer called the hypodermis which is made up of fat and connective tissue.

[0044] As used herein, the term “topically” and “topical” refers to application of the compositions of the present invention to the surface of the skin and mucous membranes.

[0045] “Topical application” or “topical administration” refers to the delivery of a composition, for treating conditions of the epidermis or dermis, wherein the topical composition is applied to the skin and acts locally and does not have a systemic effect. Topical administration of a drug may often be advantageously applied in, for example, the treatment of various skin disorders.

[0046] As used herein the terms “topical formulations” and “topical compositions” refer to formulations or compositions that may be applied to skin or mucous membranes. Topical formulations or compositions may, for example, be used to confer therapeutic benefit to a patient or cosmetic benefit to a consumer. Such topical formulations or compositions may be provided in the form of a cream, foam, gel, lotion, or ointment.

[0047] The term “Transcutol P” refers to the compound described as diethylene glycol monoethyl ether or 2-(2-ethoxyethoxy)ethanol.

[0048] The terms “treat,” “treated,” or “treating” as used herein refers to therapeutic treatment, cosmetic treatment and/or prophylactic or preventative measures, wherein the object is to prevent, reduce, eliminate or slow down (lessen) an undesired physiological condition, disorder or disease, or to obtain beneficial or desired clinical results (e.g. decrease acne, comedones, pimples, or breakouts). For the purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms; diminishment of the extent of the condition, disorder or disease; stabilization (i.e., not worsening) of the state of the condition, disorder or disease; delay in onset or slowing of the progression of the condition, disorder or disease; amelioration of the condition, disorder or disease state; and remission (whether partial or total), whether detectable or undetectable, or enhancement or improvement of the condition, disorder or disease. Treatment includes eliciting a clinically significant response without excessive levels of unwanted side effects.

[0049] The term “wart” refers to the small, rough, and hard growths that are similar in color to the rest of the skin caused by caused by infection with a type of human papillomavirus (HPV). A number of types exist including: common warts, plantar warts, filiform warts, and genital warts.

[0050] The term “vitiligo” refers to a skin condition characterized by patches of the skin losing their pigment. The patches of skin affected become white and usually have sharp margins. The hair from the skin may also become white.

[0051] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as molecular weight, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present invention.

[0052] Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0053] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability.

[0054] Methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid (RVT-501) is a phosphodiesterase type 4 inhibitor. Phosphodiesterase type 4 inhibitors, commonly referred to as a PDE4 inhibitors, are drugs used to block the degradative action of phosphodiesterase 4 (PDE4) on cyclic adenosine monophosphate (cAMP). The PDE4 family of enzymes are the most prevalent PDE in immune cells.

[0055] Alopecia areata (AA) is a T-cell-mediated autoimmune disease and has been the focus of much research, many animals models have been developed which have illuminate autoimmune pathogenesis and immunotherapeutic strategies. Gilhar, et al. Autoimmun Rev. 2016 July; 15(7): 726-735. Currently, no treatments have been approved for AA and effective therapeutic options are limited. Apremilast is an oral inhibitor of phosphodiesterase 4 that regulates production of pro- and anti-inflammatory cytokines. Pilot studies of apremilast have demonstrated that apremilast has a beneficial preventative effect by reducing the production of pro-inflammatory cytokines and retention of hair. Keren, et al. Letters to the Editor/ Journal of Dermatological Science 77 (2015): 71-81. Accordingly, PDE4 inhibitors, such as methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid (RVT-501), can be effective to treat AA.

[0056] Vitiligo is a common pigment disorder characterized by acquired development of white macules on the skin due to the loss of functioning melanocytes in the skin, the hair, or both. Vitiligo is linked to a dysregulated immune system governed by a proinflammatory cytokine network, involving local and systemic chronic inflammatory processes. Melanocytes cultured from vitiligo patient skin samples have been shown to express high levels of cytokines including IL-6 and IL-17. The use of apremilast, an oral PDE4 inhibitory has shown to be helpful in the treatment of vitiligo. Huff and Gottwald, Case Reports in Dermatological Medicine, Volume 2017, 3 pages, https://doi.org/10.1155/2017/2386234. Accordingly, PDE4 inhibitors, such as methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid (RVT-501), can be effective to treat vitiligo.

COMPOUND OF FORMULA (I)

[0057] In some embodiments, the compound represented by the formula (I) is methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid (RVT-501) having the structure:

The compound and methods of making such compound are further described in U.S. Patent Nos. 7,939,540 and 8,530,654, which are each hereby incorporated by reference in its entirety.

[0058] Optical Isomers— Diastereomers— Geometric Isomers — Tautomers.

Compounds described herein may contain an asymmetric center and may thus exist as enantiomers. Where the compounds according to the invention possess two or more asymmetric centers, they may additionally exist as diastereomers. Embodiments herein include all such possible stereoisomers as substantially pure resolved enantiomers, racemic mixtures thereof, as well as mixtures of diastereomers. The formulas are shown without a definitive stereochemistry at certain positions. Embodiments herein include all stereoisomers of such formulas and pharmaceutically acceptable salts thereof. Diastereoisomeric pairs of enantiomers may be separated by, for example, fractional crystallization from a suitable solvent, and the pair of enantiomers thus obtained may be separated into individual stereoisomers by conventional means, for example by the use of an optically active acid or base as a resolving agent or on a chiral HPLC column. Further, any enantiomer or diastereomer of a compound of the general formula may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration. Embodiments described herein include all isomers of the compound of formula (I) disclosed herein, such as a geometric isomer, an optical isomer, a stereoisomer, or a tautomer, and an isomeric mixture. Embodiments herein include both the racemic form and the optically active form. Embodiments further include a single crystal form or a mixture thereof. Moreover, embodiments herein also include an amorphous form, an anhydrate, and a hydrate form of the compound. Furthermore, embodiments herein also include metabolites, salts, hydrates, and pro-drugs of the compounds disclosed herein.

[0059] In some embodiments, a salt of compounds described herein may include an inorganic acid salt, an organic acid salt, an inorganic basic salt, an organic basic salt, an acidic or basic amino acid salt or the like. In some embodiments, the inorganic acid salt may include hydrochloride, hydrobromide, sulfate, nitrate, phosphate or the like. In some embodiments, the salt may be selected from a hydrochloride, hydrobromide, sulfate, or phosphate. In some embodiments, the organic acid salt may include acetate, succinate, fumarate, maleate, tartrate, citrate, lactate, stearate, benzoate, methanesulfonate, ethanesulfonate, p-toluenesulfonate, or benzenesulfonate. In some embodiments, the salt may be methanesulfonate or p-toluenesulfonate.

[0060] In some embodiments, the inorganic basic salt may include: alkaline metal salts such as a sodium salt or a potassium salt; alkaline-earth metal salts such as a calcium salt or a magnesium salt; aluminum salts; ammonium salts, or the like. In some embodiments, the organic basic salt may include a diethylamine salt, a diethanolamine salt, a meglumine salt, an N,N'-dibenzylethylenediamine salt, or the like.

[0061] In some embodiments, the acidic amino acid salt may include aspartate and glutamate. In some embodiments, the basic amino acid salt may include an arginine salt, a lysine salt, an ornithine salt or the like.

TOPICAL FORMULATIONS

[0062] In some embodiments, the active ingredient is methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid (RVT-501):

[0063] Embodiments herein are directed to topical compositions comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0064] In certain embodiments, a topical composition comprises methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.7% by weight, PEG 400 at 76.51% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, and butylated hydroxytoluene at 0.1% by weight.

[0065] In certain embodiments, a topical composition comprises a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0066] In certain embodiments, a topical composition comprises a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight.

[0067] Embodiments herein are directed to a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid and pharmaceutically acceptable topical excipients, wherein, within about 12 hours following administration of the topical composition, the 90% confidence interval for the ratio of means (population geometric means based on log- transformed data) of the AUC of the topical composition is within 80-125% of the AUC of any one the foregoing topical compositions and the 90% confidence internal for the ratio of means of the Cmax of the topical composition is within 70-143% of the Cmax of the same foregoing topical composition. [0068] Embodiments herein are directed to a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein the skin concentration of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 1 pg/g to about 100 pg/g within about 12 hours following administration of the topical composition.

[0069] Embodiments herein are directed to a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein the skin concentration of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 0.005 pM to about 1 pM within 12 hours of administration within about 12 hours following administration of the topical composition.

[0070] Embodiments herein are directed to a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein about 10% to about 100% of cellular Phosphodiesterase 4 (PDE4) is inhibited within 12 hours following administration of the topical composition.

[0071] The topical compositions of the present invention may be formulated by those skilled in the art as liquids, toners, solutions, sprays, emulsions, moisturizers, sunscreens, creams, lotions, masks, suspensions, triturates, gels, jellies, pastes, foams, ointments, shampoos, adhesives, serums, treated clothes or pads and the like. In some embodiments the topical composition is formulated as eye drops, as ear drops, or as a composition which can be applied to the surface of the tooth.

[0072] In embodiments described herein, the topical compositions may be applied to the skin by any means known in the art including, but not limited to, by an aerosol, spray, pump-pack, brush, swab, or other applicator. The applicator may provide either a fixed or variable metered dose application such as a metered dose aerosol, a stored-energy metered dose pump or a manual metered dose pump.

[0073] In embodiments described herein, the topical composition is formulated as to be applied to a site one time a day or multiple times per day. METHODS OF USING THE TOPICAL FORMULATIONS

[0001] Embodiments herein are directed to methods of treating a disease (as described herein) comprising administering a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein the skin concentration of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 1 pg/g to about 100 pg/g within about 12 hours following administration of the topical composition.

[0074] Embodiments herein are directed to methods of treating a disease (as described herein) comprising administering a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein the skin concentration of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 0.005 pM to about 1 pM within 12 hours of administration within about 12 hours following administration of the topical composition.

[0075] Embodiments herein are directed to methods of treating a disease (as described herein) comprising administering a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight and pharmaceutically acceptable topical excipients, wherein about 10% to about 100% of cellular Phosphodiesterase 4 (PDE4) is inhibited within 12 hours following administration of the topical composition.

[0076] Embodiments herein are directed to methods of a treating skin condition in a patient in need thereof comprising topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0077] In certain embodiments, a topical composition comprises methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.7% by weight, PEG 400 at 76.51% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, and butylated hydroxytoluene at 0.1% by weight.

[0078] In certain embodiments, a topical composition comprises a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0079] In certain embodiments, a topical composition comprises a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight.

[0080] In certain embodiments, the method of treating a skin condition in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0081] In certain embodiments, the method of treating a skin condition in a patient in need thereof comprises topically applying a topical composition wherein methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.7% by weight, PEG 400 is at 76.51% by weight, di ethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propylcellulose is at 1.75% by weight, and butylated hydroxytoluene is at 0.1% by weight.

[0082] In certain embodiments, the method of treating a skin condition in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0083] In certain embodiments, the method of treating a skin condition in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, di ethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight.

[0084] In certain embodiments, the method of treating a skin condition in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid and pharmaceutically acceptable topical excipients, wherein 90% confidence interval for the ratio of means (population geometric means based on log- transformed data) of the AUC of the topical composition is within 80-125% of the AUC of any one the foregoing topical compositions and the 90% confidence internal for the ratio of means of the Cma_x of the topical composition is within 70-143% of the Cma_x of the same foregoing topical composition.

[0085] In certain embodiments, the skin condition being treated in a patient in need thereof is selected from the group consisting of dermatitis, psoriasis, itchy skin, acne, inflammation and redness of the skin, disorders associated with sebaceous glands, oily skin, dry skin, rosacea, bums, disorders affecting the palms or soles, genetic disorders of the skin, warts, vitiligo, alopecia areata, and any combination thereof. In some embodiments, dermatitis is selected from the group consisting of atopic dermatitis, contact dermatitis, allergic contact dermatitis, irritant contact dermatitis, stasis dermatitis, seborrheic dermatitis, chronic dermatitis, eczema, and any combination thereof. In some embodiments, psoriasis is selected from the group consisting of plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, erythrodermic psoriasis, and any combination thereof. In some embodiments, itchy skin is selected from the group consisting of pruritus, prurigo, pityriasis rubra pilaris, lichen simplex chronicus, lichen planus, and any combination thereof. In some embodiments, acne is selected from the group consisting of acne vulgaris, cystic acne, inflammatory acne, non-inflammatory acne, and any combination thereof. In some embodiments, inflammation and redness of the skin is selected from the group consisting of seborrheic dermatitis, urticaria eczema, hives, seborrheic eczema, and any combination thereof. In some embodiments, disorders associated with sebaceous glands is selected from the group consisting of acne, follicular hyperkeratinization, sebostasis, sebaceous adenomas, sebaceous hyperplasia, excess sebum production, seborrhea, sebaceoma, sebaceous carcinoma, seborrheic dermatitis, sebaceous cysts, and any combination thereof. In some embodiments, oily skin is seborrhea. In some embodiments, dry skin is selected from the group consisting of sebostasis, ichthyosis, xerosis, and any combination thereof. In some embodiments, bums is sunburn. In some embodiments, disorders affecting the palms or soles is selected from the group consisting of palmoplantar pustulosis, exfoliative keratolysis, and any combination thereof. In some embodiments, genetic disorders of the skin is Darier’s disease.

[0086] In some embodiments, the method of treating atopic dermatitis in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxyl glycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0087] In certain embodiments, a method of treating atopic dermatitis in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.7% by weight, PEG 400 at 76.51% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxyl glycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, and butylated hydroxytoluene at 0.1% by weight.

[0088] In certain embodiments, a method of treating atopic dermatitis in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxyl glycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0089] In certain embodiments, a method of treating atopic dermatitis in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxyl glycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight. [0090] In some embodiments, the method of treating vitiligo in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0091] In certain embodiments, a method of treating vitiligo in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.7% by weight, PEG 400 at 76.51% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, and butylated hydroxytoluene at 0.1% by weight.

[0092] In certain embodiments, a method of treating vitiligo in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0093] In certain embodiments, a method of treating vitiligo in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight.

[0094] In some embodiments, the method of treating alopecia areata in a patient in need thereof comprises topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300. In some embodiments, methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition. In some embodiments, benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition. In some embodiments, PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition. In some embodiments, diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition. In some embodiments, oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition. In some embodiments, hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition. In some embodiments, butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition. In some embodiments, PEG 300 is at a concentration of about 5% to about 65% by weight of the topical composition.

[0095] In certain embodiments, a method of treating alopecia areata in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.7% by weight, PEG 400 at 76.51% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, and butylated hydroxytoluene at 0.1% by weight.

[0096] In certain embodiments, a method of treating alopecia areata in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 65% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxylglycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 12.21% by weight.

[0097] In certain embodiments, a method of treating alopecia areata in a patient in need thereof comprises topically applying a topical composition comprising methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid at 1.0% by weight, benzyl alcohol at 2.0% by weight, PEG 400 at 45% by weight, diethylene glycol monoethyl ether at 15% by weight, oleoyl polyoxyl glycerides at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene at 0.1% by weight, and PEG 300 at 32.12% by weight.

[0098] In embodiments described herein, the method is directed to applying a topical composition once per day. In embodiments described herein, the method is directed to applying a topical composition multiple times per day. In some embodiments, the topical composition is applied two times per day, three times per day, four times per day, or five times per day. In some embodiments, the topical composition is applied one time in the morning and one time in the evening. In some embodiments, the topical composition is applied every 12 hours, every 11 hours, every 10 hours, every 9 hours, every 8 hours, every 7 hours, every 6 hours, every 5 hours, every 4 hours, every 3 hours, every 2 hours, or every hour.

[0099] In embodiments described herein, the method is directed to applying a topical composition to multiple sites on the skin of the body. For example, the topical composition may be applied over large areas of skin prophylactically or the topical composition may be applied to particular sites in need of treatment. In some embodiments, the topical composition is applied to the skin as a liquid, toner, solution, spray, emulsion, moisturizer, sunscreen, cream, lotion, mask, suspension, triturate, gel, jelly, paste, foam, ointment, shampoo, adhesive, serum, treated cloth or pad. In some embodiments, the topical composition is applied to the eyes as eye drops, placed in the ear canal as ear drops or to the surface of the tooth.

METHODS OF DETECTING SERUM LEVELS OF METHYL N-[3-(6,7-

DIMETHOXY-2-METHYLAMINOQUINAZOLIN-4-

YL)PHENYL] TEREPHTHALAMIC ACID AND ITS METABOLITE

[00100] Embodiments herein are directed to methods of treating a condition in a patient comprising administering a topical composition comprising methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid, and analyzing the level of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid and a metabolite in the patient’s blood. In embodiments, the metabolite is 4-((3-(6,7- dimethoxy-2-(methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid.

[00101] Embodiments herein are directed to methods of treating a condition in a child comprising administering a topical composition comprising methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid, and analyzing the level of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid and a metabolite in the child’s blood. In embodiments, the metabolite is 4-((3-(6,7-dimethoxy-2- (methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid.

[00102] In embodiments, the child is less than 18 years old, less than 15 years old, less than 12 years old, less than 10 years old, less than 5 years old, less than 3 years old, less than 2 years old, or less than 1 year old. In embodiments, the child is an infant. In embodiments, the child weighs less than 50 pounds, less than 40 pounds, less than 30 pounds, less than 20 pounds, or less than 10 pounds.

[00103] Embodiments herein are directed to methods of monitoring levels of a drug and a metabolite in a patient’s blood during treatment comprising administering a topical composition of the drug, collecting the patient’s blood, and analyzing the level of the drug and a metabolite in the blood. In embodiments, the drug is methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid. In embodiments, the metabolite is 4-((3-(6,7-dimethoxy-2-(methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid.

[00104] In embodiments, the level of drug and/or metabolite in the child or patient’s blood may determine a treatment recommendation, wherein a level of drug and/or metabolite in the patient’s blood is within an acceptable limit may result in the recommendation to continue drug treatment, whereas a level of drug and/or metabolite in the patient’s blood outside of an acceptable limit may result in the discontinuation of the drug treatment or a change in the amount of drug treatment applied.

[00105] Embodiments herein are directed to methods of treating a skin condition in a patient in need thereof comprising: a) topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, b) collecting about 10 pL to about 1 mL of a blood sample from the patient, c) spotting the blood sample onto a dried blood spot card, and d) analyzing the blood sample for a level of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid and 4-((3-(6,7-dimethoxy-2-(methylamino)quinazolin-4- yl )phenyl )carb amoyl)b enzoi c aci d . [00106] In embodiments, the patient is an infant or a child, and the volume of blood collected is about 1 mL, about 500 pL, about 100 pL, about 50 pL, about 40 pL, about 30 pL, about 25 pL, about 20 pL, about 15 pL, or about 10 pL.

[00107] Embodiments herein are directed to methods of detecting methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid and 4-((3-(6,7- dimethoxy-2-(methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid comprising: a) collecting about 10 pL to about 1 mL of a blood sample from a patient, b) spotting the blood sample onto a dried blood spot card, c) punching a about 3mm to about 10 mm disc out of the dried blood spot card and processing the blood sample, d) analyzing the processed blood sample using UPLC-MS/MS (Ultra Performance Liquid Chromatography- tandem Mass Spectrometry), and e) quantifying an amount of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid and 4-((3-(6,7-dimethoxy-2- (methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid in the blood sample.

[00108] In embodiments, the volume of blood collected is about 1 mL, about 500 pL, about 100 pL, about 50 pL, about 40 pL, about 30 pL, about 25 pL, about 20 pL, about 15 pL, or about 10 pL.

[00109] In embodiments, the disc punched out from the dried blood spot card is about 3 mm, about 4 mm, about 5 mm, about 6 mm, about 7 mm, about 8 mm, about 9 mm, or about 10 mm.

[00110] In embodiments, the amount of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid quantified from the blood sample is from about 1 mg/mL to about 200 ng/mL. In embodiments, the amount of methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid quantified from the blood sample is 3 ng/mL. In embodiments, the amount of methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid quantified from the blood sample is 160 ng/mL.

[00111] In embodiments, the amount of 4-((3-(6,7-dimethoxy-2- (methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid quantified from the blood sample is from about 1 mg/mL to about 200 ng/mL. In embodiments, the amount of 4-((3- (6,7-dimethoxy-2-(methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid quantified from the blood sample is 3 ng/mL. In embodiments, the amount of 4-((3-(6,7-dimethoxy-2- (methylamino)quinazolin-4-yl)phenyl)carbamoyl)benzoic acid quantified from the blood sample is 160 ng/mL. EXAMPLES

Example 1: Formulations

[00112] Table 1 provides the compositions for gel formulations and the justifications for each ingredient.

Table l:Gel Formulations

* May include solution, ointment, cream, etc.

Example 2: Increased Permeation of RVT-501

[00113] Unexpectedly it was found that the ratio of the PEG400 to Transcutol P increased the permeation of the RVT-501 into the skin. It appears this effect is less related to Transcutol P’s effects as a penetration enhancer and more to its effects as a solvent for RVT- 501. RVT-501 dissolves in both PEG400 and Transcutol P and prefers to stay dissolved in each individual solvent versus going into the skin. In one of the ratio’s tested, the two solvents appear to antagonize each other, shifting the thermodynamics in favor of increased skin penetration.

[00114] Further, RVT-501 is highly soluble in Transcutol P and thus an interpretation of the unusual finding of seeing enhanced skin penetration at lower levels of this penetration enhancer could also be attributed to the Transcutol P “holding on” to the RVT-501 and hindering its release into the skin.

Example 3: Evaluating Topical Bioavailabilitv

[00115] Dermatopharmacokinetic (DPK) Studies

[00116] The dermatopharmacokinetic (DPK) approach is comparable to a blood, plasma, urine PK approach applied to the stratum corneum. DPK encompasses drug concentration measurements with respect to time and provides information on drug uptake, apparent steady-state levels, and drug elimination from the stratum corneum based on a stratum corneum concentration-time curve.

[00117] For antiacne drug products, target sites are the hair follicles and sebaceous glands. In this setting, the drug diffuses through the stratum corneum, epidermis, and dermis to reach the site of action. The drug may also follow follicular pathways to reach the sites of action. The extent of follicular penetration depends on the particle size of the active ingredient if it is in the form of a suspension. Under these circumstances, the DPK approach is still expected to be applicable because studies indicate a positive correlation between the stratum corneum and follicular concentrations.

[00118] Application and Removal of Test and Reference Products: The treatment areas are marked using a template without disturbing or injuring the stratum comeum/skin. The size of the treatment area will depend on multiple factors including drug strength, analytical sensitivity, the extent of drug diffusion, and exposure time. The stratum corneum is highly sensitive to certain environmental factors. To avoid bias and to remain within the limits of experimental convenience and accuracy, the treatment sites and arms should be randomized. Uptake, steady-state, and elimination phases, as described in more detail below, may be randomized between the right and left arms in a subject. Exposure time points in each phase may be randomized among various sites on each arm. The test and reference products for a particular exposure time point may be applied on sites to minimize differences. Test and reference products should be applied concurrently on the same subjects according to a SOP that has been previously developed and validated. The premarked sites are treated with predetermined amounts of the products (e.g., 5 mg/sq cm) and covered with a nonocclusive guard. Occlusion is used only if recommended in product labeling. Removal of the drug product is performed according to SOPs at the designated time points, using multiple cotton swabs or Q-tips with care to avoid stratum corneum damage. In case of certain oily preparations such as ointments, washing the area with a mild soap may be needed before skin stripping. If washing is carried out, it should be part of an SOP.

[00119] Sites and Duration of Application: The BA/BE study should include measurements of drug uptake into the stratum corneum and drug elimination from skin. Each of these elements is important to establish bioavailability and/or bioequivalence of two products, and each may be affected by the excipients present in the product. A minimum of eight sites should be employed to assess uptake/elimination from each product. The time to reach steady state in the stratum corneum should be used to determine timing of samples. For example, if the drug reaches steady-state in three hours, 0.25, 0.5, 1 and 3 hours posttreatment may be selected to determine uptake and 4, 6, 8 and 24 hours may be used to assess elimination. A zero time point (control site away from test sites) on each subject should be selected to provide baseline data. If the test/reference drug products are studied on both forearms, randomly selected sites on one arm may be designated to measure drug uptake/steady-state. Sites on the contralateral arm may then be designated to measure drug elimination. During drug uptake, both the excess drug removal and stratum corneum stripping times are the same so that the stratum corneum stripping immediately follows the removal of the excess drug. In the elimination phase, the excess drug is removed from the sites at the steady-state time point, and the stratum corneum is harvested at succeeding times over 24 hours to provide an estimate of an elimination phase.

[00120] Collection of Sample: Skin stripping proceeds first with the removal of the first 1-2 layers of stratum corneum with two adhesive tapes strip/disc applications, using a commercially available product (e.g., D-Squame, Transpore). These first two tape-strip(s) contain the generally unabsorbed, as opposed to penetrated or absorbed, drug and therefore should be analyzed separately from the rest of the tape-strips. The remaining stratum corneum layers from each site are stripped at the designated time intervals. This is achieved by stripping the site with an additional 10 adhesive tape-strips. All ten tape strips obtained from a given time point are combined and extracted, with drug content determined using a validated analytical method. The values are generally expressed as amounts/area (e.g., ng/cm²) to maintain uniformity in reported values. Data may be computed to obtain full drug concentration-time profiles, Cmax-ss, T_max-ss, and AUCs for the test and reference products.

[00121] Procedure for Skin Stripping:

[00122] To assess drug uptake: Apply the test and/or reference drug products concurrently at multiple sites. After an appropriate interval, remove the excess drug from a specific site by wiping three times lightly with a tissue or cotton swab. Using information from the pilot study, determine the appropriate times of sample collection to assess drug uptake. Repeat the application of adhesive tape two times, using uniform pressure, discarding these first two tape strips. Continue stripping at the same site to collect ten more stratum corneum samples. Care should be taken to avoid contamination with other sites. Repeat the procedure for each site at other designated time points. Extract the drug from the combined ten skin strippings and determine the concentration using a validated analytical method. Express the results as amount of drug per square cm treatment area of the adhesive tape.

[00123] To assess drug elimination: Apply the test and reference drug product concurrently at multiple sites chosen based on the results of the pilot study. Allow sufficient exposure period to reach apparent steady-state level. Remove any excess drug from the skin surface as described previously, including the first two skin strippings. Collect skin stripping samples using ten successive tape strips at time intervals based on the pilot study and analyze them for drug content.

[00124] Metrics and Statistical Analyses: A plot of stratum corneum drug concentration versus a time profile should be constructed to yield stratum corneum metrics of C_max, T_max and AUC. The two one-sided hypotheses at the a= 0.05 level of significance should be tested for AUC and C_max by constructing the 90 percent confidence interval (Cl) for the ratio between the test and reference averages. Individual subject parameters, as well as summary statistics (average, standard deviation, coefficient of variation, 90% Cl) should be reported. For the test product to be BE, the 90 percent Cl for the ratio of means (population geometric means based on log-transformed data) of test and reference treatments should fall within 80-125 percent for AUC and 70- 143 percent for Cmax.

[00125] In vivo Dermal Open Flow Microperfusion

[00126] In dermal open-flow microperfusion (dOFM), a thin, hollow tube is inserted just under the skin surface, running through a section of the skin a few inches wide and then exiting. A liquid similar to body fluid is injected into the tubing; a portion of the tube under the skin is porous, so any drug that has been applied and absorbed through the skin's outer layer enters the flowing liquid, which is then collected for analysis. dOFM can reliably measure the changing amounts of drug in the skin after topical application of a dermatological drug product.

Example 4: Clinical Study Evaluating the Effectiveness of RVT-501 in Treating Alopecia Areata

[00127] A randomized placebo-controlled pilot study will be performed to evaluate efficacy of RVT-501 in subjects with Alopecia Areata. The primary outcome measure will be the percentage of patients achieving 50% or greater improvement in their severity of Alopecia Tool (SALT) score (SALT50) at week 24 compared to Baseline. The secondary outcome measures will be: 1) the proportion of subjects achieving an alopecia areata Physician's Global Assessment (aaPGA) score of 3 or above at Weeks 24 (0, no regrowth; 1, <25% of regrowth; 2, 25%-49% of regrowth; 3, 50%-74% of regrowth; 4, 75%-99% of re growth; 5, 100% of regrowth), 2) the proportion of subjects achieving an alopecia areata Physician's Global Assessment (aaPGA) score of 3 or above at Weeks 48 (0, no regrowth; 1, <25% of regrowth; 2, 25%-49% of regrowth; 3, 50%-74% of regrowth; 4, 75%-99% of re growth; 5, 100% of regrowth), 3) the percentage change from Baseline in the Alopecia Areata Symptom Impact Scale (AASIS) at Weeks 24, 4) the percentage change from Baseline in the Alopecia Areata Symptom Impact Scale (AASIS) at Weeks 48, 5) the percentage change from baseline in the Alopecia Areata Quality of Life questionnaire (AA-QoL) at Weeks 24, 6) the percentage change from baseline in the Alopecia Areata Quality of Life questionnaire (AA-QoL) at Weeks 48, 7) the semiquantitative score using SALT subclasses (0, no hair loss; 1, <25% hair loss; 2, 25%-49% hair loss; 3, 50%-74% hair loss; 4, 75%-99% hair loss; 5, 100% hair loss) at week 24, and 8) the semiquantitative score using SALT subclasses (0, no hair loss; 1, <25% hair loss; 2, 25%-49% hair loss; 3, 50%-74% hair loss; 4, 75%-99% hair loss; 5, 100% hair loss) at week 48. Example 5: Clinical Study Evaluating the Effectiveness of RVT-501 in Treating Vitiligo

[00128] A randomized placebo-controlled pilot study or a split body study will be performed to evaluate efficacy of RVT-501 in subjects with vitiligo. The primary outcome measure will be the proportion of responders to RVT-501 at week 32 compared to baseline or control body region at week 16.

[00129] The secondary outcome measures will be:

[00130] 1) determining Body Surface Area (BSA) at weeks 32 and 48, using one hand unit, which encompasses the palm plus the volar surface of all the digits, is approximately 1% of the total body surface area and is used as a guide to estimate each body region.

[00131] 2) determining the VASI score (Vitiligo Area and Severity Index) at weeks 32 and 48, using one hand unit, which encompasses the palm plus the volar surface of all the digits, is approximately 1% of the total body surface area and is used as a guide to estimate the baseline percentage of vitiligo involvement in each body region. The body is divided into five separate and mutually exclusive regions: hands, upper extremities (excluding hands), trunk, lower extremities (excluding feet), and feet. The axillary region is included with the upper extremities while the buttocks and inguinal areas are included with the lower extremities. The extent of residual depigmentation is expressed by the following percentages: 0, 10%, 25%, 50%, 75%, 90%, or 100%. At 100% depigmentation, no pigment is present; at 90%, specks of pigment are present; at 75%, the depigmented area exceeds the pigmented area; at 50%, the depigmented and pigmented areas are equal; at 25%, the pigmented area exceeds the depigmented area; at 10%, only specks of depigmentation are present.

[00132] 3) determining VETF score (Vitiligo European Task Force) score weekly up to 64 weeks. The VETF is a validated scoring system that assesses 3 dimensions of the disease (extent, staging, and spreading/progression). (1) the extent of vitiligo will be estimated as the percentage of vitiligo involvement of 5 body sites. (2) Stage of vitiligo will be assessed as 0 (normal pigmentation), 1 (incomplete depigmentation), 2 (complete depigmentation), 3 (partial hair whitening [<30%]), and 4 (complete hair whitening). (3) Spreading of vitiligo will be scored as 0 (stable disease), -1 (observed ongoing subclinical repigmentation), and +1 (additional patches in a given area or observed ongoing subclinical depigmentation). The VETF score calculated as follows: VETF Extent or Staging or Spreading = Sum of all specific values for that category from all body sites (% of Area affected for Extent; 0-20 for Staging; -5 to +5 for Spreading). [00133] 4) assessing the Dermatology Life Quality Index weekly up to 64 weeks. The DLQI is a simple 10-question validated questionnaire that has been used in over 40 different skin conditions, and its use has been described in over 1000 publications including many multinational studies. The DLQI is the most frequently used instrument in studies of randomized controlled trials in dermatology.

[00134] 5) determining the Visual Analogue Scale (VAS) weekly up to 64 weeks. The VAS is commonly used as the outcome measure in research studies. A VAS for satisfaction is a horizontal line of 100-mm long. At the beginning and at the end, there are two descriptors representing extremes of satisfaction (i.e. no satisfaction and extreme satisfaction). The patients rate their satisfaction by making a vertical mark on the 100-mm line. The measurement in millimeters is converted to the same number of points ranging from 0 to 100 points. The exact question will be "Are you satisfied with your study treatment?"

6) recording the Number of Adverse Events weekly up to 64 weeks. Safety analyses will be performed on the safety population. Safety will be evaluated by tabulations of adverse events (AEs) and will be presented with descriptive statistics at each visit. AEs will be coded using the CTCAE, Common Terminology Criteria for Adverse Events, V 4.0. The CTCAE v4.0 AE terms are MedDRA's LLTs (Lowest Level Terms) which are based on their MedDRA (System Organ Class). The number and percentage of subjects experiencing an AE/SAE will be stratified by system organ class, or a preferred term, and/or severity of the adverse event, and recorded and tabulated overall.

Example 6: A dried blood spot assay with UPLC -MS/MS for the simultaneous determination of E6005. a phosphodiesterase 4 inhibitor and its metabolite in human blood

[00135] E6005, a novel phosphodiesterase 4 inhibitor, is currently under clinical development for the treatment of atopic dermatitis. To support pediatric clinical trials, dried blood spots assay for the simultaneous determination of E6005 and its main metabolite, ER- 392710 (Mil), has been developed using ultra-performance liquid chromatography with tandem mass spectrometry. E6005 and Mil in 25 pL blood spotted onto FTA™ DMPK-C cards were extracted by simple protein precipitation with water/acetonitrile (1:1, v/v), and then chromatographed on a reversed phase column under gradient elution. The mass transitions m/z 473.1^-163.0 for E6005 and m/z 459.1^-149.0 for Mil were monitored in a positive ion electrospray ionization mode. E6005 and Mil were quantifiable from 1 to 200 ng/mL on dried blood spots. Accuracy and precision in the intra- and inter-batch reproducibility were within the acceptance criteria recommended by the bioanalytical guidelines. Impacts on the assay accuracy by hematocrit and blood spot volume were evaluated and results showed hematocrit impacted the analytes’ accuracy. Various stability assessments including possible conversion of E6005 to Mil were thoroughly performed. The method was successfully applied to determine E6005 and Mil levels in blood samples in support of a pediatric clinical trial.

[00136] Phosphodiesterase 4 (PDE4) is expressed on various inflammatory cells and considered to play a critical role in the inflammatory disorders including atopic dermatitis. E6005 potently inhibited human PDE4 with an IC50 of 2.8 nM and also demonstrated efficacy in mice and humans, thus E6005 is considered as a promising drug for the treatment of atopic dermatitis. Atopic dermatitis is one of autoimmune diseases and a number of children and infants are suffering from. Although it is important to monitor drug concentrations in children and infants, the volume of blood sampling is limited. Dried blood spots (DBS) is a technique of micro-samplings in which small aliquots of whole blood samples were spotted onto filter paper for analysis of drugs’ levels and has been applicable for the analysis of a wide spectrum of drugs. DBS has many advantages over conventional plasma assay. It is less laborious in sample preparation of DBS compared to plasma based assay in which blood samples collected were centrifuged to obtain plasma samples for the assay. In addition, DBS requires smaller volume of blood (less than 100 pL) than conventional blood sampling (typically more than 1 mL) in typical plasma based assays.

[00137] In vitro metabolism studies demonstrated that E6005 was metabolized to various metabolites including Mil and clinical studies showed that systemic exposure of Mil in plasma was comparable to or more than that of E6005. Therefore, in a human DBS assay as well, a simultaneous assay method for the determination of E6005 and Ml 1 has been developed and validated.

Materials and Methods

[00138] Materials: E6005 and Mil were synthesized at Eisai Co., Ltd. (Ibaraki, Japan). ER-497652 and ER-497653 used as an internal standard (IS) for E6005 and Mil, respectively, were synthesized at Sekisui Medical Co., Ltd. (Ibaraki, Japan). Blank human whole blood with EDTA-2K as an anticoagulant was obtained from volunteers in Eisai Co., Ltd. with written consent. Blank human plasma was prepared by centrifuging aliquots of whole blood obtained or commercially available one was purchased from Biopredic International (Saint Gregoire, France). High-performance liquid chromatography (HPLC) grade acetonitrile, methanol, distilled water, and ammonium formate as well as a special grade formic acid were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). All other chemicals used were of analytical grade. FTAD DMPK-C blood spots cards and equipment used for disc punching including a punching device, Harris Micro Punch 3.0 mm, and cutting mat, Harris cutting mat, were purchased from GE Healthcare (Buckinghamshire, UK). Silica-gel desiccant and polyethylene bag for storing DBS cards were purchased from Toyotakako Co., Ltd. (Aichi, Japan) and Asahi Kasei home products Co. (Tokyo, Japan), respectively.

[00139] Assay conditions: The analytical conditions of E6005 and Mil in DBS were the same as those used for the validated assay in plasma. Briefly, an Acquity system (Waters, MA, USA) coupled with triple quadrupole mass spectrometer Quattro Premier (Waters) was used as an ultra-performance liquid chromatography (UPLC) with tandem mass spectrometry (ULPC-MS/MS). E6005, Mil, and IS were eluted with the mobile phase consisting of (A) water/acetonitrile/1 mol/L ammonium formate (950:50:5, v/v/v) and (B) water/acetonitrile/1 mol/L ammonium formate (100:900:5, v/v/v) and chromatographed on an Acquity UPLC BEH C18 column (2.1 mm x 100 mm, 1.7 pm, Waters) maintained at 40 °C. The gradient program is as follows: a linear increase of mobile phase (B) from 5% to 95% for 4.0 min, then an isocratic elution of 95% (B) for 0.5 min, followed by having the system equilibrated with 5% (B) for 1.5 min. The flow rate was 0.25 mL/min to 4.5 min then increased to 0.3 mL/min for equilibrium.

[00140] The optimized mass spectrometer conditions in the multiple reaction monitoring were 370 °C for desolvation temperature, 125 °C for source temperature, and 1.3 kV for capillary voltage, 65 V for cone voltage, and -55 eV for collision energy. The mass transition m/z (precursor ion®product ion) 473.1^-163.0, m/z 459.1^-149.0, m/z 477.2®167.0, and m/z 463.2®153.0 were monitored for E6005, Mil, IS of E6005, and IS of Mil, respectively.

[00141] Preparation of calibration and quality control samples: A mixture of stock solutions of E6005 and Mil in methanol (each 100 pg/mL as free base) was diluted with acetonitrile/methanol (1:1, v/v) to prepare working standard solutions. By fortifying working solutions to blank naive whole blood (hematocrit: ca. 45%), calibration samples were prepared at concentrations of 1, 2, 10, 20, 80, 100, 160, and 200 ng/mL for both E6005 and Mil. Fresh blank whole blood was used to prepare calibration samples otherwise stated. A working solution of the IS in acetonitrile/methanol (1:1, v/v) was prepared in the similar way as described above (200 ng/mL). The working solutions were stored below -20°C and used within 181 days in which stability was confirmed. Quality control (QC) samples including the lower limit of quantification (LLOQ), low QC (LQC), middle QC (MQC), and high QC (HQC), were prepared at concentrations of 1, 3, 30, and 160 ng/mL blood with designated hematocrit values. Blood samples with varying hematocrits were prepared by mixing plasma and blood cells with the nominal ratios of 80:20 to 30:70 (v/v). Accurate hematocrit values determined using a hematology analyzer (AD VIA 120, Siemens, Munich, Germany) were 19.3, 26.9, 36.2, 46.7, 49.1, 51.8, 57.5, and 63.6% for the nominal hematocrit of 20% (80:20), 30% (70:30), 40% (60:40), 50% (50:50), 53% (47:53), 56% (44:56), 60% (40:60), and 70% (30:70), (plasma/blood cells, v/v), respectively. Aliquots (25 pL) of blood samples (calibration samples and QC samples) were spotted onto the center of circle of FTA™ DMPK-C cards using a calibrated pipette to prepare DBS. The cards were allowed to dry at room temperature for at least 2 h. QC samples used for the long-term stability assessment were stored at designated temperature in a sealed polyethylene bag containing Silica-gel desiccant.

[00142] Sample extraction procedures: DBS discs (i.d. 3 mm) were punched out at the center of spots using the punching device, Harris Micro Punch, into tubes for extraction. A 10 pL aliquot of the IS working solution (200 ng/mL) was spiked and then the analytes were extracted by 100 pL acetonitrile/water (1:1, v/v). After vigorously vortexing, samples were centrifuged (15700xg, 1 min) at 4°C to obtain supernatants for injection. A 10 pL aliquot of supernatants was injected to the UPLC -MS/MS system.

Method validation

[00143] Linearity: Punched discs spotted with calibration samples (1 - 200 ng/mL for both E6005 and Mil) were extracted and assayed to determine inaccuracy (relative error, RE) at each concentration across 8 assay runs. Inaccuracy of determined E6005 and Mil at each concentration should be within ±15% (±20% was allowed for the LLOQ). Imprecision as relative standard deviation (RSD) was also calculated and checked whether % RSD was 15% or less (20% or less was allowed for the LLOQ).

[00144] Specificity: Discs spotted with blank human blood from six individuals were extracted to check if there were any endogenous peaks interfering with assay of the analytes. Interfering peak areas should be less than 20% for E6005 and Ml 1 while 5% for IS of those of LLOQ samples.

[00145] Intra- and inter-batch reproducibility: Inaccuracy and imprecision of E6005 and Ml 1 were determined using QC samples (LLOQ, LQC, MQC, and HQC) in the intra- and inter-assay batch. Five replicates per concentration were assessed for the intra-batch reproducibility, and intra-batch evaluation was repeated across three batches for the inter batch reproducibility. The acceptance criteria for inaccuracy and imprecision were within ±15% and 15%, respectively (±20% for inaccuracy and 20% for imprecision are allowed for the LLOQ samples).

[00146] Extraction recovery and matrix effect: Extraction recovery of E6005 and Mil from DBS discs was assessed at three concentrations (3, 30, and 160 ng/mL, three replicate/concentration), while recovery of the IS from the system was determined at 60 ng/mL. Extraction recovery of the analytes was determined by dividing the peak area of the analytes spiked to blank blood prior to extraction by that spiked after extraction (reference samples) taking the differences in areas between discs for extraction and blood spots into account while extraction recovery of the IS was determined just by comparison of peak area between the extracted samples and reference ones without any correction. Blood spot areas were calculated by nr², where r is radius of spots determined by a ruler.

[00147] Matrix factors were evaluated by dividing peak area of reference samples from six individuals by that of neat solution with identical concentrations. Matrix factors were determined for the analytes of interest (E6005 and Mil) at 3 ng/mL and the corresponding IS at 160 ng/mL. IS-corrected matrix factors of E6005 and Mil were calculated by dividing matrix factor of E6005 and Mil by that of the corresponding IS. The % RSD of the IS-corrected matrix factor should be within 15%.

[00148] Effect of blood spot volume, hematocrit, and punching location: As potential impacts on assay accuracy by blood spot volume, hematocrit, and punching location are unique to DBS -associated bioanalytical method validation studies, these parameters were also assessed. To assess potential impacts by blood spot volume, various volume of QC samples (10, 20, 25, 30, and 40 pL) at low (3 ng/mL) and high (160 ng/mL) concentrations were spotted onto the circle of DBS cards, and then center-punched discs were assayed in three replicates against the calibration samples with the fixed volume (25 pL). Acceptable blood spot volumes should have inaccuracy < ±15%.

[00149] Effects of hematocrit on the assay of E6005 and Mil were evaluated using blood samples with varying hematocrit values (19.3% to 63.6%) at low (3 ng/mL) and high (160 ng/mL) concentrations with the other conditions fixed (25 pL spot volume and center- punching). DBS discs spiked by blood with varying hematocrit were assayed in three replicates against calibration samples prepared from naive blood (hematocrit: 45.1%). Potential relationship between spot area and inaccuracy of QC samples was evaluated. Impacts of hematocrit were considered negligible when the inaccuracy was not greater than ±15%.

[00150] Potential impacts of punching locations in discs were assessed for the following four locations of the spot with the other condition fixed (25 pL spot volumes and 45.1% hematocrit): upper right, lower right, upper left, and lower left. Low (3 ng/mL) and high (160 ng/mL) concentrations were evaluated. Discs punched out from four locations were assayed with the center-punched disc of calibration samples. No impact of punching location was suggested when the inaccuracy was within ±15%.

[00151] Carryover: Two types of carryover assessments should be evaluated in bioanalytical methods using DBS-based assays; one is carryover derived from repetitive sample injection via UPLC, a typical validation parameter in the method validation, and the other is DBS-specific spot-to-spot carryover mainly derived from punching devices by repetitive punching of discs. The carryover in the UPLC was assessed by injecting blank samples just after upper limit of quantification (ULOQ) samples. The other possible carryover caused by repetitive punching was investigated by punching discs with blank samples just after the ULOQ samples using a punching device without any wash. Peak areas of any interferences in blank samples should be less than 20% and 5% of the LLOQ sample for the analytes of interest and the IS, respectively.

[00152] Stability: Following stability of E6005 and Mil in DBS was assessed at low and high concentrations using LQC and HQC samples (three replicates/concentration): bench-top stability for 7 days at room temperature, long-term frozen stability for 160 days at room temperature and below -15°C, and processed sample stability for 85 h at 4°C. To investigate impacts of high humidity on the stability, bench-top stability test was performed at room temperature with relative humidity of ca. 80%-84%. Samples were considered stable when % bias from the nominal concentrations was within ±15%.

[00153] As a part of the stability assessment, possible conversion of E6005 to Ml 1 was also investigated using HQC samples in which only E6005 was fortified. After designated times, formed Mil concentrations were determined and percentage of conversion was calculated by dividing Mil concentrations by E6005 concentrations on the molar concentration basis.

[00154] Shelf-life of refrigerated blood: As it is sometimes a challenge that fresh blood samples are available to prepare calibration or QC samples, it is of interest to know whether or not refrigerated blood can be used. The shelf-life of refrigerated blood was assessed by assaying QC samples at low (3 ng/mL) and high (160 ng/mL) concentrations in three replicates prepared from refrigerated blood for seven days against calibration samples prepared from fresh blood. The refrigerated blood can be used when % bias from the nominal concentrations was within ±15%.

[00155] Clinical application: A clinical study was performed in which E6005 ointment containing 0.05% or 0.2% was topically applied twice a day for two weeks to pediatric subjects. Blood samples were obtained at 1- and 2-week post-dose as well as subsequent 7-day follow-up period in collection tubes with K2-EDTA as an anticoagulant, thereafter put on ice as soon as possible to reduce possible conversion of E6005 to Mil. Details on sample handling at the clinics were clarified in a lab manual; a 25 pL aliquot blood sample was spotted onto the center of circle of DBS cards (four replicates per sample) at clinics, then dried at room temperature for at least 2 h. DBS cards with desiccants were placed in zip lock bags and stored frozen below -20°C until shipment to a bioanalytical laboratory. Samples were stored below -15°C at the laboratory until they were subjected to sample processing for the determination of E6005 and Mil concentrations in DBS.

Results and Discussion

Method Development

[00156] Blood spotting is one of the crucial steps in the DBS method to ensure accurate determination, thus in the method development, some abuses on blood spotting were investigated. Typically blood samples with drugs of interest were spotted by one drop per spot with a pipette. As an abuse of double drop may be possible at clinics, DBS with the double drop of blood samples (each 15 pL aliquots) was processed and concentrations of E6005 and Mil were determined against calibration samples with single blood drop (30 pL aliquots) to ensure whether the RE (%) was within ±15%. The RE (%) of double drop samples was -4.6% and 3.7% for E6005 and Mi l, respectively, suggesting minimal impacts of double drop of blood samples as long as the total volume is comparable. The laboratory manual indicates that pipettes should be kept just above the DBS paper not touched when spotting, however blood spot may be performed with pipettes touched on cards; the % RE of E6005 and Mil was 4.3% and 9.7%, respectively, indicating minimal impacts by pipette’ touching on card when spotting.

[00157] Extraction procedure focused on selecting appropriate extraction solvents: acetonitrile, acetonitrile/water (8:2, v/v), acetonitrile/water (1:1, v/v), methanol, methanol/water (8:2, v/v), and methanol/water (1:1, v/v). Although minimal extraction was noted with acetonitrile, other solvents showed similar extraction efficiency. Less endogenous peaks in chromatograms led to the selection of 50% acetonitrile rather than pure organic solvents or higher organic solvent containing solvents.

[00158] One possible issue to be addressed in the method development is lower sensitivity in DBS method due to lower volume of matrix compared to traditional plasma assay. Given the target LLOQ (1 ng/mL blood), increases in punching spot area were tested for whether higher sensitivity could be achieved. Other than 3 mm diameter disc punching, 6 mm diameter punching was assessed and peak intensity of the analytes was increased 3 to 4 folds which was comparable to the theoretical increase (4 folds).

Method Validation

[00159] Linearity and selectivity: E6005 and Ml 1 were quantifiable ranging from 1 to 200 ng/mL with acceptable % inaccuracy and imprecision at all the concentrations tested (Table 2). Calibration curves were consistent among assay batches with minimal variability of the slope (8.2% and 8.6% for E6005 and Mil, respectively).

Table 2: Linearity of E6005 and Mil in human dried blood spots

Inaccuracy and imprecision as the relative standard deviation (RSD) were calculated from 8 analytical runs. [00160] Accuracy and precision: Intra- and inter-batch accuracy and precision were assessed at four levels (LLOQ, LQC, MQC, and HQC) and results are shown in Table 3. The inaccuracy as % RE and imprecision as % RSD for E6005 and Ml 1 were within ±7.0% and 9.6% in the intra-batch test and within ±8.0% and 15.7% (at the LLOQ) in the inter-batch test, respectively. These results were within the acceptance criteria recommended by the bioanalytical guidelines from LIS Food and Drug Administration and European Medicines Agency. No dilution integrity was assessed since it was highly unlikely that the analytes’ levels exceeded the LiLOQ (200 ng/mL) in clinical studies.

Table 3: Intra- and inter-batch reproducibility of E6005 and Mil in human dried blood spots

Quality Nominal Inaccuracy (%) Imprecision (%) control concentration E6005 Mil E6005 Mil (ng/mL)

Intra-batch (n=5/batch)

LLOQ Ϊ 4.0 7.0 9.6 4.7

LQC 3 -1.3 1.0 4.4 5.9

MQC 30 6.3 5.0 6.3 3.2

HQC 160 -1.3 -1.3 6.3 4.4

Inter-batch (n=15, n=5/batch, 3 batches in total)

LLOQ ϊ TO 8Ό 140 15.7

LQC 3 0.7 1.0 7.3 7.6

MQC 30 1.3 0.3 6.3 6.0

HQC 160 1.9 1.9 5.5 5.5

[00161] Extraction recovery and matrix effect: Table 4 shows extraction recoveries of the analytes of interest and the IS. Extraction recoveries of E6005 and Mil at low, middle, and high concentrations were 79.2%-86.7% and 73.3%-87.5%, respectively, by taking the differences in disc areas between extracted and spotted into account. The recovery of the IS was 93.7% for E6005 and 96.9% for Mil. Extraction recoveries of E6005 and Mil were consistent across the concentrations tested. Relatively lower extraction of the analytes than the IS was attributable to differences in fortifying neat solution in the system, where the analytes were spotted onto cards before extraction while the IS was just fortified after extraction of the analytes.

Table 4: Mean extraction recovery of E6005 and Mil and the internal standards (IS) Samples Nominal % Mean recovery concentration E6005 Mil

(ng/mL)

LQC 3 81.5+1.5 80.9+4.4

MQC 30 79.2+2.7 73.3+2.3

HQC 160 86.7+6.5 87.5+7.5

IS 60 93.7+4.9 96.9+4.5

Data represent the mean ± standard deviation of three replicates for the analytes at each level and nine replicates for the IS.

[00162] Matrix effects of the analytes and the IS were evaluated using blood from six individuals. Neither ion suppression nor ion enhancement were observed for both analytes and the IS with matrix factor ranging from 92.5% to 100.3%. The IS-normalized matrix factor was almost unity ranged from 93.2% to 99.2% with CV of 2.2% for E6005 and 2.1% for Mil, indicating no matrix effects (Table 5).

Table 5: Matrix effects of E6005 and Mil in human dried blood spots from six individuals

Matrix effects were evaluated at 5 ng/mL for E6005 and Ml 1.

[00163] Impacts of blood spot volume, hematocrit, and punching location: Possible impacts of blood spot volume were investigated by spotting blood samples (10 to 40 pL) containing E6005 and Mil at low and high concentrations. The % RE of both E6005 and Mil was within the acceptance criteria (< ±15%) when 10 to 30 pL blood was spotted. On the other hand, inaccuracy of Mil at 40 pL spotting was slightly higher than the criteria (16.3%). These results suggest that blood spot volume was ensured at least up to 30 pL.

[00164] Impacts of hematocrit were also assessed using blood samples with varying hematocrit (19.3% - 63.6%). The % RE of E6005 and Mil was within < ±15% at hematocrit ranging from 26.9% to 51.8%. On the other hand, inaccuracy negatively biased at the hematocrit 19.3% while positively biased at the hematocrit 57.5% and 63.6%. It would be explained by varying viscosity of blood; the diffusion of blood on DBS cards would be higher for blood with smaller hematocrit, thus resulted in larger blood spot area. As the same size of discs were punched out regardless of blood spot area, larger blood spot area leads to lower concentrations of analytes, and vice versa. The impact of hematocrit on the assay accuracy of E6005 and Mil was not clinically significant given that hematocrit of subjects determined in a clinical study ranged from 0.33 to 0.47.

[00165] As punching locations of a disc in a spot may impact the assay of E6005 and Mil, the % RE of the analytes in four different punching locations (upper, lower, right, and left of spots) was compared to that in the center of spots. Discs punched out from all the peripheral location showed % RE within ±15% when concentrations were determined against calibration samples punched from the center of disc, indicating minimal impact of disc punching locations.

[00166] Carryover: No carryover derived from repetitive sample injection was detected in chromatograms of blank samples injected just after ULOQ samples. No DBS- specific device-oriented carryover was also noted. [00167] Stability: Results of the stability assessment of E6005 and Mil in DBS are shown in Table 6. A bench-top stability test demonstrated that E6005 and Mil were stable for 160 days at ambient temperature. Long-term frozen stability was assessed below -15°C and confirmed stable up to 160 days. Stability of E6005 and Mil in processed samples was confirmed for 85 h when stored at 4°C. No impacts of high humidity on the stability were also ensured for 7 days at ambient temperature with relative humidity of ca. 80%-84%. Possible conversion of E6005 to Mil was evaluated by fortifying only E6005 in blood and formed Mil levels were assessed (Table 7). The conversion of E6005 in the long-term stability test was slightly higher at room temperature (2.2%) compared to that stored below - 15°C (1.2%). However, the conversion was not so different between 7 and 160 days even when stored at room temperature (1.0% and 2.2% for 7 and 160 days, respectively). The minimal conversion of E6005 to Mil was not clinically significant given minimal exposure of E6005 after dermal application (1.65 ng/mL at the maximum).

Table 6: Stability of E6005 and Mil in human dried blood spots

Quality control samples at low (3 ng/mL) and high (160 ng/mL) levels were assayed in triplicates and the relative error was calculated from the mean. Percent bias was calculated against nominal concentrations.

Table 7: Conversion of E6005 to Mil in human dried blood spots

[00168] Blood-to-plasma partition and shelf-life of refrigerated blood: The blood-to- plasma partition (B/P) of E6005 and Mil was determined by assaying concentrations of E6005 and Ml 1 in whole blood samples and plasma samples prepared from blood samples by centrifugation. The B/P of E6005 was 0.690 and 0.669 at 3 and 160 ng/mL, respectively, while that of Mil was 0.594 and 0.574 at 3 and 160 ng/mL, respectively, suggesting that no concentration-dependent B/P was observed. The average B/P of two levels was 0.679 for E6005 and 0.584 for Mi l.

[00169] The shelf-life of refrigerated whole blood was assessed by evaluating the inaccuracy of QC samples from “aged blood” fortified with E6005 and Mil at low and high levels. The % RE of E6005 was 7.7% and -1.9% at 3 and 160 ng/mL, respectively, and that of Mil was 7.3% and -3.8% at the low and high concentrations, respectively. These results suggest that aged blood stored refrigerated for seven days can be used in preparing calibration samples and QC samples.

[00170] Clinical application: Concentrations of E6005 and Mil in blood were determined in support of a pediatric clinical trial in which E6005 was topically applied. A total of 147 DBS samples were assayed according to the method described above and all the samples except one were below the LLQ. The maximum level was 1.65 ng/mL for E6005 while that of Mil was below the LLOQ. These results demonstrated that the systemic exposures of E6005 and Ml 1 were minimal when E6005 was applied topically to children, which was similar to findings in adults. The % RE of all the calibration samples and QC samples assayed with post-dose samples was within ±15%, indicating accurate determination of E6005 and Ml 1 in sample assay.

[00171] Conclusions

[00172] Results in the validation study demonstrated that the developed DBS method with UPLC -MS/MS for the simultaneous determination of E6005 and Mil in human whole blood is simple, selective, and reproducible. The validated method has been successfully applied to clinical studies in which blood E6005 levels in pediatrics were accurately determined using only 25 pL blood.

Example 7: Topical Pharmacokinetic and Skin Penetration data of RVT-501 Gel Formulation

[00173] A 7 day repeat dose study was performed on 10 healthy volunteers. F3 Gel composition as provided in Table 1. On days 1-3, RVT-501 0.5% ointment was applied to 2 sites twice daily (0.5% BSA each) and RVT-501 1.0% gel was applied to 2 sites twice daily (0.5% BSA each). On day 3, 12 hours post morning application, two 2mm skin biopsies were collected. On days 4-7, RVT-501 0.5% ointment was applied to 1 site twice daily (0.5% BSA each) and RVT-501 1.0% gel was applied to 1 sitel twice daily (0.5% BSA each). On day 7, 12 hours post morning application, two 2mm skin biopsies were collected. Follow up visits were conducted days 7-10 post last application. [00174] Safety Data: 3 Subjects reported AEs; 5 Subjects reported No AEs. All AEs were Grade 1/Mild (Headache (2), sore throat (unrelated), upper respiratory infection, biopsy site pain, burning sensation at application site, 1 mild AE of burning sensation at application site (0.5% ointment), 1 report of barely perceptible skin irritation reported (1% gel) on Day 1 at 1 hour post application. All AEs were resolved.

[00175] Results: RVT-501 concentrations were measurable in all 40 skin samples (10 subjects x 4 samples per subject). Mil metabolite concentrations were below the limit of quantification in all 40 skin samples.

[00176] Drug level was measured in homogenized skin samples 12 hours post application. FIG. 1 provides free fraction skin concentration (pg/g) at day 3 for the 1% RVT- 501 Gel and 0.5% RVT-501 Ointment, as well as at day 7 for the 1% RVT-501 Gel and 0.5% RVT-501 Ointment. Table 8 provides mean, median and ratio data; data assumes a skin density of about 1 g/ml, molecular weight of RVT-501 is 472.5 pg/pmol, and human skin protein binding is 99.4%.

Table 8: Drug Level in Homogenized Skin 12 Hours Post-Dose

[00177] BPS Bioscience PDE4 IC50 cell-based functional assay and Cerep IC50 purified enzyme binding assay were utilized to determine the percent inhibition in each subject sample. FIG. 2 provides free fraction skin concentration (mM) at day 3 for the 1% RVT-501 Gel and 0.5% RVT-501 Ointment, as well as at day 7 for the 1% RVT-501 Gel and 0.5% RVT-501 Ointment. Table 9 provides median, ratio data, and percent inhibition for each assay. Table 9: Comparison of Free Fraction of RVT-501 to PDE4 IC50

* Assumes a Hill Coefficient of 1

[00178] Conclusions: PK data suggests that application of 1.0% RVT-501 Gel leads to approximately 2.0-fold to 4.1-fold higher skin RVT-501 concentrations than application of 0.5% RVT-501 Ointment when applied to healthy human skin.

Claims

1. A topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300.

2. The topical composition of claim 1, wherein methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition.

3. The topical composition of claim 1, wherein benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition.

4. The topical composition of claim 1, wherein PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition.

5. The topical composition of claim 1, wherein di ethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition.

6. The topical composition of claim 1, wherein oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition.

7. The topical composition of claim 1, wherein hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition.

8. The topical composition of claim 1, wherein butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition.

9. The topical composition of claim 1, wherein PEG 300 is present and is at a concentration of about 5% to about 65% by weight of the topical composition.

10. The topical composition of claim 1, wherein methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.7% by weight, PEG 400 is at 76.51% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propylcellulose is at 1.75% by weight, and butylated hydroxytoluene is at 0.1% by weight.

11. The topical composition of claim 1, wherein methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.0% by weight, PEG 400 is at 65% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propyl cellulose at 1.75% by weight, butylated hydroxytoluene is at 0.1% by weight, and PEG 300 is at 12.21% by weight.

12. The topical composition of claim 1, wherein methyl N-[3-(6,7- dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.0% by weight, PEG 400 at 45% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propylcellulose is at 1.75% by weight, butylated hydroxytoluene is at 0.1% by weight, and PEG 300 is at 32.12% by weight.

13. A method of treating a skin condition in a patient in need thereof comprising topically applying a topical composition comprising a therapeutically effective amount of methyl N-[3-(6,7-dimethoxy-2-methylaminoquinazolin-4- yl)phenyl]terephthalamic acid, benzyl alcohol, PEG 400, diethylene glycol monoethyl ether, oleoyl polyoxylglycerides, hydroxyl propylcellulose, butylated hydroxytoluene, and optionally PEG 300.

14. The method of claim 13, wherein methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at a concentration of about 0.01% to about 5% by weight of the topical composition.

15. The method of claim 13, wherein benzyl alcohol is at a concentration of about 0.1% to about 5% by weight of the topical composition.

16. The method of claim 13, wherein PEG 400 is at a concentration of about 30% to about 80% by weight of the topical composition.

17. The method of claim 13, wherein diethylene glycol monoethyl ether is at a concentration of about 10% to about 60% by weight of the topical composition.

18. The method of claim 13, wherein oleoyl polyoxylglycerides is at a concentration of about 1% to about 5% by weight of the topical composition.

19. The method of claim 13, wherein hydroxyl propylcellulose is at a concentration of about 0.5% to about 5% by weight of the topical composition.

20. The method of claim 13, wherein butylated hydroxytoluene is at a concentration of about 0.01% to about 1.0% by weight of the topical composition.

21. The method of claim 13, wherein PEG 300 is present and is at a concentration of about 5% to about 65% by weight of the topical composition.

22. The method of claim 13, wherein methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.7% by weight, PEG 400 is at 76.51% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propylcellulose is at 1.75% by weight, and butylated hydroxytoluene is at 0.1% by weight.

23. The method of claim 13, wherein methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.0% by weight, PEG 400 is at 65% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propylcellulose at 1.75% by weight, butylated hydroxytoluene is at 0.1% by weight, and PEG 300 is at 12.21% by weight.

24. The method of claim 13, wherein methyl N-[3-(6,7-dimethoxy-2- methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is at 1.0% by weight, benzyl alcohol is at 2.0% by weight, PEG 400 at 45% by weight, diethylene glycol monoethyl ether is at 15% by weight, oleoyl polyoxylglycerides is at 2.94% by weight, hydroxyl propyl cellulose is at 1.75% by weight, butylated hydroxytoluene is at 0.1% by weight, and PEG 300 is at 32.12% by weight.

25. The method of claim 13, wherein a skin concentration of methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 1 pg/g to about 100 pg/g within 12 hours following administration of the topical composition.

26. The method of claim 13, wherein a skin concentration of methyl N-[3- (6,7-dimethoxy-2-methylaminoquinazolin-4-yl)phenyl]terephthalamic acid is about 0.005 mM to about 1 mM within 12 hours of administration within 12 hours following administration of the topical composition.

27. The method of claim 13, wherein about 10% to about 100% of cellular Phosphodiesterase 4 (PDE4) is inhibited within 12 hours following administration of the topical composition.

28. The method of claim 13, wherein the skin condition is selected from the group consisting of dermatitis, psoriasis, itchy skin, acne, inflammation and redness of the skin, disorders associated with sebaceous glands, oily skin, dry skin, rosacea, burns, disorders affecting the palms or soles, genetic disorders of the skin, warts, vitiligo, alopecia areata, and any combination thereof.

29. The method of claim 28, wherein dermatitis is selected from the group consisting of atopic dermatitis, contact dermatitis, allergic contact dermatitis, irritant contact dermatitis, stasis dermatitis, seborrheic dermatitis, chronic dermatitis, eczema, and any combination thereof.

30. The method of claim 28, wherein psoriasis is selected from the group consisting of plaque psoriasis, guttate psoriasis, inverse psoriasis, pustular psoriasis, erythrodermic psoriasis, and any combination thereof.

31. The method of claim 28, wherein itchy skin is selected from the group consisting of pruritus, prurigo, pityriasis rubra pilaris, lichen simplex chronicus, lichen planus, and any combination thereof.

32. The method of claim 28, wherein acne is selected from the group consisting of acne vulgaris, cystic acne, inflammatory acne, non-inflammatory acne, and any combination thereof.

33. The method of claim 28, wherein inflammation and redness of the skin is selected from the group consisting of seborrheic dermatitis, urticaria eczema, hives, seborrheic eczema, and any combination thereof.

34. The method of claim 28, wherein disorders associated with sebaceous glands is selected from the group consisting of acne, follicular hyperkeratinization, sebostasis, sebaceous adenomas, sebaceous hyperplasia, excess sebum production, seborrhea, sebaceoma, sebaceous carcinoma, seborrheic dermatitis, sebaceous cysts, and any combination thereof.

35. The method of claim 28, wherein oily skin is seborrhea.

36. The method of claim 28, wherein dry skin is selected from the group consisting of sebostasis, ichthyosis, xerosis, and any combination thereof.

37. The method of claim 28, wherein burns is sunburn.

38. The method of claim 28, wherein disorders affecting the palms or soles is selected from the group consisting of palmoplantar pustulosis, exfoliative keratolysis, and any combination thereof.

39. The method of claim 28, wherein genetic disorders of the skin is Darier’s disease.

40. The method of claim 13, wherein said skin condition is atopic dermatitis.

41. The method of claim 13, wherein said skin condition is vitiligo.

42. The method of claim 13, wherein said skin condition is alopecia areata.