WO2016044750A1 - Topical formulations of growth factors - Google Patents

Abstract

Provided herein are compositions, methods of treatment using such compositions, and methods of making such compositions wherein the compositions comprise a large molecule. The compositions, and devices and kits comprising such compositions provide new and useful improvements as alternatives for addressing diseases, disorders and symptoms thereof in a subject.

Description

Topical Formulations of Growth Factors

The transdermal patch segment of the pharmaceutical industry currently commands a relatively small share of the rapidly growing global drug delivery market ($21.5 billion in 2010, projected growth to $31.5 billion by 2015). Approved patches are currently available for drugs with properties that lend themselves to passive permeation across the skin when applied topically such as estrogen, nicotine, nitroglycerin, scopolamine, fentanyl and clonidine.

To date, patch-based penetration enhancement formulations have been limited to delivering small chemical drugs below a molecular weight size of 500 daltons (the so-called "rule of 500") due to the physical constraints of effectively transporting large water-soluble compounds across the thick, keratin-rich, armor- like outer layer of skin (stratum corneum). The underlying cellular layers that comprise the viable epidermis, also present a rate-limiting barrier to transdermal drug delivery after successful penetration of the stratum corneum. The barrier to drug diffusion across the epidermis is probably the presence of tight junctions. Removal of the full epidermis increased skin permeability by 1-2 orders of magnitude depending on the molecule delivered.

Currently, there is a need for improved delivery of large molecular weight molecules across skin and systemically in order to address diseases, disorders, and symptoms thereof in patients in need.

SUMMARY

Provided herein are compositions, methods of treatment using such compositions, and methods of making such compositions wherein the compositions comprise a large molecule. The compositions, and devices and kits comprising such compositions provide new and useful improvements as alternatives for addressing diseases, disorders and symptoms thereof in a subject.

One aspect is a composition comprising propylene glycol (PG), poloxamer 188 (P188), laurocapram (LP) and human growth hormone (hGH). The composition is that wherein:

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 49:39: 12 (by weight);

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 55:43:2 (by weight);

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 56:44:0 (by weight);

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 50:50: 12 (by weight);

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 50:50:2 (by weight);

the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 50:50:0 (by weight);

the hGH is lyophilized hGH;

the hGH is present in a range of 0.1-10 mg/50 μΙ_;

further comprising insulin-like growth factor 1 (IGF-1);

the IGF-1 is present in a range of 0.1-10 mg/50 μΙ_;

the composition herein that is devoid of azone. Another aspect is a method of promoting growth in a subject comprising administration of any composition herein to the subject. In other aspects, the method is that wherein:

the administration is topically;

the hGH is administered at a range of 0.1-10 mg/50 μΙ_/0ΠΊ²;

the administration of hGH is via a patch attached to the subject for up to 168 hours;

the administration of hGH is via a patch attached to the subject for up to 24 hours;

the administration of hGH is via a patch attached to the subject for up to 168, 144, 120, 96, 72, 48, or 24 hours;

the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 24 hours after initial administration;

the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 48 hours after initial administration;

the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 72 hours after initial administration;

the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 168, 144, 120, 96, 72, 48, or 24 hours after initial administration;

the composition of claim 1 is in solid form;

the composition of claim 1 is in gel form;

the hGH is administered continuously and consistently over at least 7 days;

the hGH is administered continuously and consistently over at least 3 days;

the hGH is administered continuously and consistently over at least 24 hours;

the hGH is administered continuously and consistently over at least 168, 144, 120, 96, 72, 48, or 24 hours; the hGH is administered such that serum hGH accumulation of at least 5-200 ng/mL is achieved over 24-168 consecutive hours (e.g., 168, 144, 120, 96, 72, 48, or 24 hours);

the hGH is administered such that serum hGH accumulation of at least 10-150 ng/mL is achieved over 24-168 consecutive hours (e.g., 168, 144, 120, 96, 72, 48, or 24 hours);

the hGH is administered such that serum hGH accumulation of at least 5-100 ng/mL is achieved over 24-168 consecutive hours (e.g., 168, 144, 120, 96, 72, 48, or 24 hours);

the hGH is administered such that serum hGH accumulation of at least 10-125 ng/mL is achieved over 24-168 consecutive hours (e.g., 168, 144, 120, 96, 72, 48, or 24 hours);

the hGH is administered at a dosage of 0.5-100 mg/wk;

the hGH is administered at a dosage of 1-50 mg/wk;

the hGH is administered at a dosage of 0.5-15 mg/wk.

Another aspect is a kit comprising a composition of herein and a device for topical administration of the composition herein. In other aspects, the kit is that wherein:

the device is a patch;

the composition herein is in solid form;

the composition herein is in gel form;

the patch further comprises an adhesive material;

it further comprises a cover material to protect the composition herein during shipping and storage;

the patch is 1 x 1 (cm x cm) to 10 x 10 (cm x cm);

the patch is 1 x 1, 2 x 2, 3 x 3, 4 x 4, 5 x 5, 6 x 6, 7 x 7, 8 x 8, 9 x 9 or 10 x 10 (cm x cm). Another aspect is a device comprising a composition herein and a patch. In other aspects, the device is that wherein: the patch comprises a backing;

the patch comprises an adhesive;

the patch comprises a laminate coated with a composition;

the patch comprises a release liner;

the patch comprises the composition herein in solid form;

the patch comprises the composition herein in gel form;

the patch is 1 x 1 (cm x cm) to 10 x 10 (cm x cm);

the patch is 1 x 1, 2 x 2, 3 x 3, 4 x 4, 5 x 5, 6 x 6, 7 x 7, 8 x 8, 9 x 9 or 10 x 10 (cm x cm). Accumulation of active agent via sustained release patch can be calculated based on the minimum steady-state flux rate at 1 ng/cm /hr. For example, at such rate then in the first 24 hrs, accumulation is: (1 ng/0.6 cm /hr) x 24 hr/2.5 ml = 16 ng/ml. After 168 hrs, accumulation is: (1 ng/0.6 cm²/hr) x 168 hr/2.5 ml = 112 ng/ml.

In an aspect, the device (e.g., patch) provides a steady-state flux (permeation) of active agent at a constant rate. So, if set at a minimum at 1 ng/ cm /hr, one obtains 24 ng/ cm in 24 hours. The rate remains the same every consecutive 24 hours (1 day) for up to 7 days. So, whether 24 or 168 hours after administration, the steady-state flux remains the same.

A surface of the device adjacent to the contact site can be between about 1 and about 100 square centimeters in size, including range of integers (e.g., 1-10, 5-50, 50-100, etc.), or any integer between about 1 and about 100. In some embodiments, the device can be between about 1 and about 25 cubic centimeters in size. The therapeutic-agent-containing formulation can include a thermo- sensitive polymer. In some embodiments, the thermo- sensitive polymer can include a poloxamer or a poloxamine. The poloxamer can be poloxamer 188.

The device (e.g., a patch) can comprise a dermoadhesive agent. The dermoadhesive agent in the therapeutic-agent-containing formulation can be selected from the group including of propylene glycol, dipropylene glycol, polyethylene glycol, glycerine, butylene glycol, glycol derivatives with glycerol esters, and non-ionizable glycol ether derivatives.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts hGH steady state-flux in a topical patch administration.

FIG. 2 depicts in vitro serum hGH levels of topical patch embodiments.

FIG. 3 depicts hGH patch dosing of the invention compared to injection.

FIG. 4 depicts atomic structure of stratum corneum.

FIG. 5-7 depict tables showing the results of a pharmacokinetic (PK) simulation model to demonstrate that hGH patch embodiments of reasonable patch dimensions, total patch dose, and maximum duration of action can achieve upper end of the range of normal random serum hGH levels within 12 hours after application for each age/gender group: a) FIG. 5 for men (5 ng/mL); b) FIG. 6 for women (10 ng/mL); and c) FIG.7 for children (20 ng/mL).

FIG. 8 depicts a table showing comparison by age/gender of simulated hGH patch PK values.

FIG. 9 depicts equations for pharmacokinetic modeling to predict patch size and duration of action.

DEFINITIONS

This disclosure can be better understood with reference to the following definitions:

As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

Unless specifically stated or obvious from context, as used herein, the term "about" is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. "About" can be understood as within 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.

As used herein, the terms "comprises," "comprising," "containing," "having," and the like can have the meaning ascribed to them under U.S. patent law and can mean "includes," "including," and the like.

Unless specifically stated or obvious from context, the term "or," as used herein, is understood to be inclusive.

The term "perforation" shall be understood to refer to a series of holes made into a material that allows easy separation of two sections of the material. The holes may be circular or may be elongated. The process of creating perforations involves puncturing the material with a tool. Perforations can be formed by a hole punch or a cutting edge that includes "nicks," (i.e., indentations) where the two sections of the material are not separated. Alternately, perforations can be made by a cutting wheel or a grinding wheel that includes nicks in the wheel's circumference.

Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 (as well as fractions thereof unless the context clearly dictates otherwise).

DETAILED DESCRIPTION OF THIS DISCLOSURE

At least one embodiment of this disclosure provides co-polymer/enhancer formulations for passive topical and transdermal delivery of large molecule drugs. Advantageously, in the preferred embodiments, the co-polymer/enhancer formulations disclosed herein are comprised of individual components generally regarded as safe (GRAS) and are thermo- sensitive, or dermoadhesive, and enhance the penetration of therapeutics across skin or other surfaces. The individual components may possess two or more of these three properties (primary, secondary and/or tertiary properties). The co-polymer/enhancer formulations can be embodied as a passive transdermal delivery patch that delivers one or more therapeutics continuously for up to 7 days or more. Also provided are therapeutic uses of the co-polymer/enhancer co-polymer/enhancer formulations for non-invasive, needle-free delivery of large peptide drugs including but not limited to hGH, IGF-1, etc.

In a preferred embodiment, the delivery system comprises co-polymers of poloxamer 188 (PI 88) and propylene glycol (PG), the penetration-enhancer laurocapram (Azone) and, optionally, other classes of penetration-enhancing compounds (including short penetration- enhancing peptides), and one or more therapeutic agents. In a preferred embodiment, the delivery formulation disclosed herein can be used for transdermal delivery of large molecule perishable drugs (e.g. peptides or proteins) useful for enhancing or promoting growth in a subject.

In a preferred embodiment, the delivery system comprises co-polymers of poloxamer 188 (PI 88) and propylene glycol, the penetration-enhancer laurocapram (Azone) and, optionally, other classes of penetration-enhancing compounds (including short penetration-enhancing peptides), and one or more therapeutic agents.

The co-polymer/enhancer formulations disclosed herein are thermo-sensitive; that is, solid at room temperature for ease of application, for example, to the skin (30-32° C) or insertion into the vaginal canal to coat the cervical transformation zone (core body temperature of 37° C), and transition (melt) to a gel or liquid phase at these physiological temperatures. The ratio of poloxamer 188, propylene glycol, laurocapram, and a specific drug in a formulation may be altered to determine the transition temperature. For example, a specific composition may determine a solid-to-gel transition at skin temperature 30-32° C, while a different composition may determine a solid-to-gel transition at core body temperature of 37° C. Furthermore, the co- polymer/enhancer formulations in gel or liquid phase adhere to and enhance large and small molecule penetration across the skin or mucosa.

Advantageously, the co-polymer/enhancer formulations disclosed herein allow for noninvasive, targeted delivery of therapeutics across the skin and mucosal surfaces. Additionally, the present co-polymer/enhancer formulations use safe, inexpensive ingredients, are easy to administer and are suitable for use in a wide range of clinical settings. The co-polymer/enhancer formulations can easily be administered by healthcare workers or by self-administration by the patient, and under conditions of extreme temperature, high humidity, poor lighting, lack of space or lack of adequate supply of electricity or water.

Co-Polymer/Enhancer Formulation for Topical Delivery of Therapeutic Compounds

One aspect of this disclosure provides co-polymer/enhancer formulations for topical delivery of therapeutics. Advantageously, the co-polymer/enhancer formulations of this disclosure are thermo-sensitive, mucoadhesive or dermoadhesive, and enhance the penetration of therapeutics across the full thickness of the skin or mucosal surfaces.

In one embodiment, the novel topical delivery formulation comprises a thermo-sensitive polymer, a mucoadhesive or dermoadhesive polymer, a penetration enhancer and, optionally, one or more therapeutic agents (e.g., large molecule therapeutic agent, growth factor).

In one embodiment, the co-polymer/enhancer formulation comprises co-polymer of one or more thermo-sensitive polymers, one or more mucoadhesive or dermoadhesive polymers and one or more penetration-enhancing agents. In a preferred embodiment, the co-polymer/enhancer formulation comprises co-polymer of poloxamer 188 and propylene glycol, the penetration- enhancer laurocapram and, optionally, one or more therapeutic agents.

In one embodiment, the co-polymer/enhancer formulation comprises one or more polymeric materials including, but not limited to, poloxamer and poloxamine. Poloxamers useful according to this disclosure include, but are not limited to, poloxamer 188, 407, 101, 105, 108, 122, 123, 124, 181, 182, 183, 184, 185, 212, 215, 217, 231, 234, 235, 237, 238, 282, 284, 288, 331, 333, 334, 335, 338, 401, 402, and 403. Poloxamines useful according to this disclosure include, but are not limited to, poloxamine 304, 504, 701, 702, 704, 707, 901, 904, 908, 1101, 1102, 1104, 1301, 1302, 1304, 1307, 1501, 1502, 1504, and 1508. In certain embodiments, the co-polymer/enhancer formulation can comprise one or more polymeric materials including, but not limited to, polylactic acid and copolymers, polyvinyl acetate, celluloses and derivatives (such as carboxymethyl cellulose, cellulose acetate, cellulose acetate propionate, ethyl cellulose, hydroxypropyl methyl cellulose, hydroxyalkyl methyl celluloses and alkyl celluloses), crosslinked dextrans, polyethylene glycol, diethylaminoethyl dextran, poly(cyanoacrylates), copolymers of PEG and PLA, poly(lactic-co-glycolic acid), poly(ortho esters) and hydrogels. Preferably, the polymeric material is pharmaceutically- acceptable, biodegradable, mucoadhesive or dermoadhesive and/or enhances the penetration of therapeutics across the skin and/or mucosal surface.

In one embodiment, the co-polymer/enhancer formulation further comprises one or more mucoadhesive or dermoadhesive agents. In one embodiment the mucoadhesive or dermoadhesive agent promotes adhesion of the co-polymer/enhancer formulation to the skin or mucosa membranes. Preferably, the mucoadhesive or dermoadhesive agent also enhances the penetration of therapeutics across the skin and/or mucosal surface.

Mucoadhesive or dermoadhesive agents useful according to this disclosure include, but are not limited to, polyols such as, propylene glycol, dipropylene glycol, polyethylene glycol, glycerine and butylene glycol; glycol derivatives with glycerol esters, such as, oleic acid esters of propylene glycol; and non-ionizable glycol ether derivatives, such as, ethoxydiglycol.

Mucoadhesive or dermoadhesive agents useful according to this disclosure, can also include polymers such as, polyethylene glycol caprylic/capric glycerides; vinyl polymers (e.g., polyhydroxyethyl acrylate, polyhydroxyethyl methacrylate, polyvinyl alcohol and polyvinyl pyrrolidone); cellulosic derivatives, such as, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose and carboxymethyl cellulose; polysaccharides, such as, alginic acid and sodium alginate.

In one embodiment, the topical delivery formulation further comprises one or more penetration enhancers. Penetration enhancers useful according to this disclosure include, but are not limited to, laurocapram, diethylene glycol, monoethyl ether, n-decyl methyl sulfoxide, dimethyl sulfoxide, dimethylacetamidedimethylformamide, sucrose monooleate, amides and other nitrogenous compounds (e.g., urea, 2-pyrrolidone, l-methyl-2-pyrrolidone, ethanolamine, diethanolamine and triethanolamine), organic acids (e.g., citric acid and succinic acid), N- methyl-2-pyrrolidine, borage oil, tetrahydropiperine (THP), alcohols (e.g., methanol, ethanol, propanol, octanol, benzyl alcohol, myristyl alcohol, cetyl alcohol, stearyl alcohol), fatty acids (e.g., oleic acid), fatty acid esters (e.g., isopropyl myristate, isopropyl palmitate), polyols (e.g., propylene glycol, polyethylene glycol, glycerol), polyethylene glycol monolaurate and lecithin.

Preferably, the co-polymer/enhancer formulation is solid or semi-solid at room temperature, begins to melt at temperatures slightly below, and completely melts at physiological temperatures. Generally, room temperature is below 30 °C, below 28 °C, below 25 °C, below 23 °C, below 20 °C, or below 18 °C.

In certain embodiments, the co-polymer/enhancer formulation melts, or begins to melt, at a temperature ranging from about 30 °C to 42 °C, 32 °C to 40 °C, 33 °C to 40 °C, 35 °C to 38 °C, or 34 °C to 37 °C. In certain embodiments, the co-polymer/enhancer formulation melts, or begins to melt, at a temperature above 30 °C, 31 °C, 32 °C, 33 °C, 34 °C, 35 °C, 36 °C, or 37 °C. In certain embodiments, the biopolymer formulation melts, or begins to melt, at a temperature below 45 °C, 44 °C, 43 °C, 42 °C, 41 °C, 40 °C, 39 °C, 38 °C, 37 °C, 36 °C, 35 °C, or 34 °C. The desired thermal property of the co-polymer/enhancer formulation can be achieved by adjusting the relative ratio (e.g., in terms of weight percentages or molar amounts) of various ingredients including, the thermo-sensitive polymeric material, the mucoadhesive agent, the penetration enhancer and/or the therapeutic agent.

In certain embodiments, the co-polymer/enhancer formulation comprises a polymeric material at a weight percentage of about 20% to about 95%, about 25% to about 90%, about 30% to about 85%, about 35% to about 80%, about 40% to about 70%, about 50% to about 90%, about 50% to about 85%, about 60% to about 80%, about 30% to about 40%, about 30% to about 50%, about 70% to about 90%, about 70% to about 85% or about 70% to about 80%.

In certain embodiments, the co-polymer/enhancer formulation comprises a mucoadhesive or deraioadhesive agent at a weight percentage of about 5% to about 90%, about 10% to about 80%, about 10% to about 70%, about 10% to about 60%, about 10% to about 50%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 20%, about 5% to about 30%, about 5% to about 20%, about 5% to about 15% or about 15% to about 30%.

In certain embodiments, the co-polymer/enhancer or co-polymer/retardant formulation comprises a penetration enhancer or penetration retardant, respectively, at a concentration ranging from about 1 to about 12 %, about 2 to about 11%, about 3 to about 10%, about 4 to about 9%, about 5 to about 8%, about 2 to about 9 %, about 3 to about 8%, about 4 to about 7%, about 2 to about 5%, or about 6 to about 7% by weight.

In certain embodiments, the co-polymer/enhancer or co-polymer/retardant formulation comprises a penetration enhancer or penetration retardant, respectively, at a concentration above 0.5%, 1%, 1.5%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25% or 30% by weight. In certain embodiments, the co-polymer/enhancer or co-polymer/retardant formulation comprises a penetration enhancer or penetration retardant, respectively, at a concentration below 70%, 60%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6% or 5% by weight.

In certain specific embodiments, the co-polymer/enhancer formulation comprises poloxamer 188 and propylene glycol at a ratio (w/w) of about 100:0, 90: 10, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90, or 0: 100. In preferred embodiments, the co-polymer/enhancer formulation comprises poloxamer 188 and propylene glycol at a ratio (w/w) of about 70:30 (37 °C or core body temperature) or 50/50 (30-32 °C or skin temperature).

In certain embodiments, the co-polymer/enhancer formulation comprises laurocapram at a concentration of about 1 to about 12 %, about 2 to about 11%, about 3 to about 10%, about 4 to about 9%, about 5 to about 8%, about 2 to about 9 %, about 3 to about 8%, about 4 to about 7%, about 2 to about 5%, or about 6 to about 7% by weight. In a preferred embodiment, the co- polymer/enhancer formulation comprises about 12% laurocapram by weight. In another preferred embodiments, the co-polymer/enhancer formulation comprises about 2% laurocapram, by weight. In a another embodiment, the co-polymer/enhancer formulation comprises about <12%, <11%, <10%, <9%, <8%, <7%, <6%, <5%,<4%, <3%, <2%, or <1%, laurocapram by weight. It is contemplated that the amount of laurocapram can be selected to be a balance between a desired flux rate and negative side effects (e.g., irritation, toxicity).

The co-polymer/enhancer formulation can be used for topical delivery of a variety of small or large therapeutic agents not previously achieved using penetration enhancers including, but not limited to, large peptides and proteins greater than about 50 amino acids, (e.g., >53 amino acids, >60 amino acids, >70 amino acids), nucleic acids, compounds with unique physicochemical structures and/or properties not considered amenable to passive transdermal or transmucosal delivery.

In one embodiment, hormones can include but not be limited to human or bovine growth hormone (hGH or bGH), or insulin-like growth factor 1 (IGF-1).

In certain embodiments, the co-polymer/enhancer formulation comprises a therapeutic agent at a concentration ranging from 0.1 mM to about 3 mM, about 0.1 mM to about 2 mM, about 0.1 mM to about 1.5 mM, about 0.5 mM to about 2 mM, or about 0.5 mM to about 1.5 mM, about 0.01 mM to about 30 mM, about 0.01 mM to about 20 mM, about 0.1 mM to about 15 mM, about 0.05 mM to about 20 mM, or about 0.05 mM to about 15 mM. Amounts of therapeutic agents incorporated into co-polymer/enhancer formulations disclosed herein can also be determined by those skilled in the art (e.g., based upon age, bioavailability of a therapeutic agent, etc.) such that the therapeutic agent is delivered to a subject in amounts that effect a therapeutic benefit to the subject.

In one embodiment the method comprises administering, to skin or mucosal surface of a subject, a co-polymer/enhancer formulation of this disclosure using any standard topical patch design or variations thereof, having in common the direct application (bio-interface) with the skin or mucosal surface. In a specific embodiment the method comprises administering, to skin or mucosal surface of a subject, a co-polymer/enhancer formulation comprising poloxamer 188 and propylene glycol, laurocapram and, optionally, one or more therapeutic agents.

The term "subject," as used herein, describes an organism, including mammals such as primates, to which treatment with the formulations according to the subject disclosure can be provided. Mammalian species that can benefit from the disclosed methods of treatment include, but are not limited to, apes, chimpanzees, orangutans, humans, monkeys; and domesticated animals such as dogs, cats, horses, cattle, pigs, sheep, goats, chickens, mice, rats, guinea pigs, and hamsters.

In certain embodiments the co-polymer/enhancer formulation of this disclosure is administered to skin or mucosal surfaces including, but not limited to, cervix, vagina, anus, rectum, eye, ear, nose, thorax, vulva, larynx, and head and neck.

Embodiments of this disclosure allow for topical delivery of therapeutics across the full- thickness skin barrier including the keratinized apical layer of skin (stratum corneum), the epidermal cell layer, and the dermis, and/or mucosa. At least one embodiment allows for topical delivery of therapeutics across non-keratinized surface of skin and/or mucosa. At least one embodiment allows for topical delivery of therapeutics into, or across, multiple layers of cervical squamous epithelial cells. At least one embodiment allows for topical delivery of therapeutics to the basal keratinocytes of skin and/or mucosa.

At least one embodiment can be used as a non-invasive topical transdermal or transmucosal delivery system (or device) applied to normal skin or mucosal surfaces to obviate the need for subcutaneous injection of therapeutic compounds. A patch for topical transdermal delivery of a composition herein can include any suitable material (e.g., transdermal system, backings, liners, membranes and tapes, and the like) well known in the transdermal delivery art. Such materials are commercially available from various sources and known in the art. See for example, those available from 3M Drug Delivery Systems, St. Paul, MN.

Other aspects useful in application of this technology and embodiments herein are described in International Patent Application No. PCT/US2014/030259, incorporated by reference. Formulations and Formulations for Topical Administration

The subject disclosure also provides for therapeutic or pharmaceutical formulations comprising the co-polymer/enhancer formulation in a form that can be combined with a pharmaceutically acceptable carrier. In a preferred embodiment the therapeutic or pharmaceutical formulation is solid at room temperature and transitions to a gel or liquid at desired physiological temperatures.

The term "carrier" refers to a diluent, adjuvant, excipient or vehicle with which the compound is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum oil such as mineral oil, vegetable oil such as peanut oil, soybean oil and sesame oil, animal oil or oil of synthetic origin.

Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol and the like. The therapeutic formulation, if desired, can also contain minor amounts of wetting, emulsifying or pH buffering agents. These formulations can take the form of creams, foam, patches, lotions, drops, sprays, gel, oils, aerosol, powders, ointment, solutions, suspensions, emulsion and the like. The formulation can be formulated with traditional binders and carriers such as triglycerides. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin. Such formulations contain a therapeutically effective amount of the therapeutic formulation, together with a suitable amount of carrier so as to provide the form for proper administration to the patient. The formulation should suit the mode of administration.

The subject disclosure also provides for the modification of the ingredient such that it is more stable once administered to a subject, i.e., once administered it has a longer time period of effectiveness as compared to the unmodified form. Such modifications are well known to those of skill in the art, e.g., microencapsulation, etc.

The amount of the therapeutic or pharmaceutical formulation of this disclosure which is effective in the treatment of a particular disease, condition or disorder will depend on the route of administration and the seriousness of the disease, condition or disorder and should be decided according to the judgment of the practitioner and each patient's circumstances.

Further, at least one embodiment provides kits containing therapeutic compositions herein. Preferably, the formulations of this disclosure are stable in a wide range of temperatures below the desired melting temperature. In one embodiment the active therapeutic agents can be reconstituted by mixing pre-measured quantities of each component immediately prior to use. The kits can include a covering package material (e.g., plastic, foil, polymer material, etc.) suitable to protect the kit and its contents (e.g., active agent, patch, device) from degradation and contamination.

The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

EXAMPLES

Following are examples that illustrate procedures for practicing this disclosure. These examples should not be construed as limiting. All percentages are by weight and all solvent mixture proportions are by volume unless otherwise noted. EXAMPLE 1

There is a need for a convenient and affordable alternative to daily subcutaneous (sc) injections for a growing incidence and prevalence of childhood- and adult-onset human growth hormone (hGH) deficiency. hGH is also called somatotropin and growth hormone- 1 (GH- 1) to distinguish it from the related protein growth hormone-2 (GH-2). We sought to develop a convenient, non-invasive and affordable transdermal patch formulation capable of achieving passive delivery of large molecule drugs such as hGH and insulin for multiple days. We report a novel platform technology, a composition, applied to the transdermal delivery of hGH (22, 128 Da). This solution was lyophilized overnight. The lyophilized powder was reconstituted in an optimized formulation to create a stock solution. The formulation consisted of about 49% poloxamer 188, about 39% propylene glycol, and about 12% laurocapram (wt./wt./wt).

Formulations containing hGH at different concentrations were freshly prepared from the stock solution immediately prior to application as 50 μΐ_^ gel to EpidermPT™ (EFT-300) tissue inserts, which are mitotically and metabolically active human skin equivalents (HSEs) when cultured in maintenance medium at 32°C. Formulations over the dose range of 0.0001- 1.0 mg/0.6 cm hGH were incubated at 32° C for 7-days. Equal volume of medium was sampled and replenished every 12 rs. hGH concentrations were measured using the Quantikine^® ELISA kit (R&D Systems) as per manufacturer' s instructions. Steady-state hGH flux (J_ss) was calculated using Pick' s first law. We observed a clear dose-response (FIG 1). We tested the feasibility of developing an effective extended- wear hGH patch capable of providing constant and consistent drug delivery to replace daily or weekly injections for therapy of growth hormone deficiency (GHD) in children and adults. Feasibility was demonstrated by using J„ from these in vitro studies along with published pharmacokinetic (PK) data for hGH in humans to perform simulations using two different PK models as described in the figures below.

FIG 1: Passive transdermal delivery of human growth hormone (hGH, somatropin) continuously over 7-days using an embodiment of a formulation. Notably, hGH has a molecular weight (MW) of 22,124 Da, which is -3.8X larger than human insulin (5,808 Da). The flux of hGH exhibits a dose-response in the range of 0.0005-0.5 mg/0.6 cm over the course of a 7-day study in a highly robust human skin equivalent model (HSE). MatTek's EpiDermFT™ (EFT- 300) tissue inserts are barrier-enhanced (i.e. stratum corneum 3X the thickness of normal human skin), full-thickness, metabolically and mitotically active HSEs that remain viable for at least 7- days when maintained in serum-containing medium in a tissue culture incubator at 32° C (skin temperature). Continuous and consistent transdermal delivery was observed, as indicated by the linear accumulation plots (same amount of delivery each day characteristic of constant steady- state flux rates). Saturation was observed at an administered dose of 0.3 mg/0.6 cm , since the accumulation curves for the 0.3 mg/0.6 cm and 0.5 mg/0.6 cm doses are superimposable, although the true saturating dose may be between 0.2-0.3 mg/0.6 cm .

FIG 2: Table showing extrapolated serum hGH levels in humans as a determinant of hGH patch dimensions or surface area (cm ) and time (1-7 days). Extrapolated serum hGH levels are from flux observed for a 0.5 mg/0.6 cm saturating dose of an embodiment of an hGH formulation in EpiDermFT™ (EFT-300) accumulation dose-response studies using 2.5 ml of maintenance medium (FIG. 1), extrapolated to 5L adult blood volume.

Shadowing (light blue) in upper left quadrant corresponds to patch size and time required to remain in the normal range of random serum levels specific for Men (< 5 ng/mL or 226 pmol/L). Shadowing (intermediate blue) in middle diagonal cells corresponds to additional patch size and time required to remain in normal range of random serum levels specific for Women: < 10 ng/mL (452 pmol/L).

Shadowing (darker blue) in right lower quadrant corresponds to patch size and time required to remain in normal range of random serum levels specific for Children 0-20 ng/mL (0- 904 pmol/L).

Shadowing (darkest blue) in right lower quadrant corresponds to patch size and time required to remain in normal range of random serum levels specific for Newborns: 5-40 ng/mL (226-1808 pmol/L).

The extrapolated serum hGH levels are derived from an in vitro tissue model that only assesses transdermal transport as a measure of accumulation of permeant as a function of time to demonstrate constant and consistent delivery (curve should be linear; slope defines dose- response). The values reflect a model of 100% drug metabolism/elimination each day since medium is removed and replaced with fresh medium every 24 hours for the duration of the 7-day study; this methodology is a requirement for creating "in sink" conditions for calculating steady- state flux from Fick's 1st Law of Diffusion.

FIG 3: Table showing extrapolated maximum hGH patch size carrying an equivalent absolute amount of hGH (upper limit) compared to weekly therapy with Genotropin® for GHD by injection.

Adult (age > 18): Assume dosing for average adult male of 70 kg

Pediatric (age < 18): Assume dosing for average 2-18 year old male of 30-70 kg, respectively. Pediatric: maximum patch size between 3 x 3 cm (7.50 mg) and 4 x 4 cm (13.3 mg) can achieve normal range of random serum levels for a week.

Adults (Weighted): maximum patch size between 2 x 2 cm (3.33 mg) and 3 x 3 cm (7.5 mg) can achieve normal range of random serum levels for Adults (Men and Women). Adults (Unweighted): maximum patch size between l x l cm (0.83 mg) and 2 x 2 cm (3.33 mg) can achieve normal range of random serum levels for Adults (Men and Women) for a week.

FIG 4: Remarkably, the complex structure of skin morphology has not been fully elucidated. It was first proposed by Wertz, Swartzendruber, Madison and Downing (1983) that the fat layer of the stratum corneum extracellular space is composed of saturated long-chain ceramides, free fatty acids and cholesterol in a 1: 1: 1 molar ratio. Norlen (2001) proposed that the fat layer exists as a single and coherent gel phase. It was not until 2012 that Norlen' s team reported the use of very high magnification cryoelectron microscopy (EM) of vitreous skin section defocus series, molecular modeling, and EM simulation to determine the atomic structure of the stratum corneum lipid matrix in situ, in its near-native state. The structure is composed of stacked bilayers of fully extended ceramides with cholesterol molecules associated with the ceramide sphingoid moiety; the free fatty acid is associated with the long-chain fatty acid moiety of the ceramide.

FIG 5-7: Tables showing the results of a pharmacokinetic (PK) simulation model to demonstrate that hGH patch embodiments of reasonable patch dimensions, total patch dose, and maximum duration of action can achieve upper end of the range of normal random serum hGH levels within 12 hours after application for each age/gender group: a) FIG. 5 for men (5 ng/mL); b) FIG. 6 for women (10 ng/mL); and c) FIG.7 for children (20 ng/mL). Setting the target levels at the upper end of the normal range is intended to enhance the robustness of the proof-of- concept PK simulations. Results are based on hGH steady-state flux observed for embodiments of hGH formulations over the dose range of 0.005-0.5 mg/0.6 cm in EpiDermFT™ (EFT-300) accumulation dose-response studies using 2.5 ml of maintenance medium (FIG 1). Pharmacokinetic calculations are based on non-compartmental analysis (NCA) when the elimination process follows first-order kinetics, as most of drugs do when they are used at their therapeutic doses (Foye's Principles of Medicinal Chemistry). The ideal simulation model of transdermal patch delivery would require use of published PK data from continuous subcutaneous infusion. We used the continuous intravenous infusion analysis function of the PKSolver 2.0 add-in PK software for Excel to simulate a NCA extravascular model using the formulas in FIG. 9.

FIG 8: Table showing comparison by age/gender of simulated hGH patch PK values. Notably, the values corresponding to the columns highlighted in red in FIG 5-7 indicate that patch dimensions are below 8 x 8 cm (upper limit of convenient size) and below 10 x 10 cm (upper limit of usage), and that the duration of action is up to 12-weeks. The latter supports the feasibility of developing an effective extended- wear hGH patch by a wide margin, certainly a 1-2 week patch that is the most practical life-span for extended-wear use.

FIG 9: Equations for pharmacokinetic modeling to predict patch size and duration of action. Although there is no steady-state level for serum hGH in normal individuals due to its wide fluctuation throughout the day, there is a range for random serum hGH measurements according to age/gender. The steady- state constant (C_ss) in equation 3 is utilized to define a hypothetical target serum steady-state level that can be achieved by constant and consistent delivery by an hGH patch in the setting of GHD. Passive transdermal delivery of large molecule drugs (e.g., IGF-1, hGH) mediated by embodiments of formulations as disclosed herein is hypothesized to occur by the synergistic effects of its components (thermosensitive polymer, dermoadhesive agent, and a chemical penetration enhancer)— all of which contribute chemical penetration enhancing properties. The inclusion of a non-ionic amphipathic triblock copolymer (poloxamer 188) suggests the possibility of the formation of "pores" in which the lipophilic polyoxypropylene core interacts with the surrounding structure of the extracellular lipid matrix of the stratum corneum (as determined above), while the hydrophilic moieties of the flanking polyoxyethylene arms are partitioned in the center of the pore. The sizes of such putative pores might differ over a range in a stochastic distribution. The size distribution of such putative pores might follow a bell-shaped curve, such that small molecule drugs "pass through" all of the pores, intermediate-sized drugs (e.g. insulin) pass through the majority of pores constituting the pores with dimensions distributed across the central portion of the bell-shaped curve, and large drugs (e.g. hGH or larger) only pass through a smaller number of large pores at the upper size distribution of the bell-shaped curve. If this is the case, one strategy for increasing the flux of large molecules is to shift the bell-shaped curve towards the upper size distribution (i.e. "shift curve to the right").

EQUIVALENTS

The functions of several elements may, in alternative embodiments, be carried out by fewer elements, or a single element. Similarly, in some embodiments, any functional element may perform fewer, or different, operations than those described with respect to the illustrated embodiment. While certain embodiments according to this disclosure have been described, this disclosure is not limited to just the described embodiments. Various changes and/or

modifications can be made to any of the described embodiments without departing from the spirit or scope of this disclosure. Also, various combinations of elements, steps, features, and/or aspects of the described embodiments are possible and contemplated even if such combinations are not expressly identified herein.

INCORPORATION BY REFERENCE

The entire contents of all patents, published patent applications, and other references cited herein are hereby expressly incorporated herein in their entireties by reference.

Claims

CLAIMS What is claimed is:

1. A composition comprising propylene glycol (PG), poloxamer 188 (P188), laurocapram (LP) and hGH.

2. The composition of claim 1, wherein the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 49:39: 12 (by weight).

3. The composition of claim 1, wherein the ratio of propylene glycol (PG), poloxamer 188 (P188), and laurocapram (LP) is 55:43:2 (by weight)

4. The composition of claim 1, wherein the ratio of propylene glycol (PG), poloxamer 188 (PI 88), and laurocapram (LP) is 56:44:0 (by weight)

5. The composition of claim 2, wherein the hGH is lyophilized hGH.

6. The composition of claim 2, wherein the hGH is present in a range of 0.1- 10 mg/50 μΙ_.

7. The composition of claim 1, further comprising IGF-1.

8. The composition of claim 5, wherein the IGF-1 is present in a range of 0.1-10 mg/50 μΙ_.

9. The composition of claim 1 that is devoid of azone.

10. A method of promoting growth in a subject comprising administration of the composition of claim 1 to the subject.

11. The method of claim 10, wherein the administration is topically.

12. The method of claim 11, wherein the hGH is administered at a range of 0.1-10 mg/50 μΙ_/οιη².

13. The method of claim 11, wherein the administration of hGH is via a patch attached to the subject for up to 168 hours.

14. The method of claim 11, wherein the administration of hGH is via a patch attached to the subject for up to 24 hours.

15. The method of claim 11, wherein the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 24 hours after initial administration.

16. The method of claim 11, wherein the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 48 hours after initial administration.

17. The method of claim 11, wherein the permeation of hGH through EpiDermFT™ is at least 1 ng/cm /hr 72 hours after initial administration.

18. The method of claim 11, wherein the composition of claim 1 is in solid form.

19. The method of claim 11, wherein the composition of claim 1 is in gel form.

20. The method of claim 11, wherein the hGH is administered continuously and consistently over at least 7 days.

21. The method of claim 11, wherein the hGH is administered continuously and consistently over at least 3 days.

22. The method of claim 11, wherein the hGH is administered continuously and consistently over at least 24 hours.

23. The method of claim 11, wherein the hGH is administered such that serum hGH accumulation of at least 5-200 ng/niL is achieved over 24-168 consecutive hours.

24. The method of claim 11, wherein the hGH is administered such that serum hGH accumulation of at least 10-150 ng/niL is achieved over 24-168 consecutive hours.

25. The method of claim 11, wherein the hGH is administered such that serum hGH accumulation of at least 5-100 ng/niL is achieved over 24-168 consecutive hours.

26. The method of claim 11, wherein the hGH is administered such that serum hGH accumulation of at least 10-125 ng/niL is achieved over 24-168 consecutive hours.

27. The method of claim 11, wherein the hGH is administered at a dosage of 0.5-100 mg/wk.

28. The method of claim 11, wherein the hGH is administered at a dosage of 1-50 mg/wk.

29. The method of claim 11, wherein the hGH is administered at a dosage of 0.5-15 mg/wk.

30. A kit comprising a composition of claim 1 and a device for topical administration of the composition of claim 1.

31. The kit of claim 30, wherein the device is a patch.

32. The kit of claim 30, wherein the composition of claim 1 is in solid form.

33. The kit of claim 30, wherein the composition of claim 1 is in gel form

34. The kit of claim 31, wherein the patch further comprises an adhesive material.

35. The kit of claim 31, further comprising a cover material to protect the composition of claim 1 during shipping and storage.

36. The kit of claim 31, wherein the patch is no larger than 10 x 10 (cm x cm).

37. A device comprising a composition of claim 1 and a patch.

38. The device of claim 37, wherein the patch comprises a backing.

39. The device of claim 37, wherein the patch comprises an adhesive.

40. The device of claim 37, wherein the patch comprises a laminate coated with the composition of claim 1.

41. The device of claim 37, wherein the patch comprises a release liner.

42. The device of claim 37, wherein the patch comprises the composition of claim 1 in solid form.

43. The device of claim 37, wherein the patch comprises the composition of claim 1 in gel form.

44. The device of claim 37, wherein the patch is no larger than 10 x 10 (cm x cm).

45. The device of claim 37, wherein the patch is 1 x 1 (cm x cm).