FORMULATIONS OF PTHrP ANALOGUES, TRANSDERMAL PATCHES THEREOF, AND USES THEREOF

SEQUENCE LISTING

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INTRODUCTION

Conventionally, osteoporosis is treated by administration of antiresorptive agents to suppress bone resorption. The most common of these treatments is oral or intravenous administration of bisphosphonates. However, an undesirable side effect of bisphosphonate administration is reduced bone formation (MacLean 2008). Anabolic agents provide an alternative to antiresorptives. The only anabolic agent currently available for treatment of osteoporosis is teriparatide (PTH (1-34), Forteo®), a recombinant form of parathyroid hormone (PTH) that acts by a mechanism that involves stimulating new bone formation (along with resorption) and reconstituting internal bone microarchitecture (Recker 2009; Dempster 2012; Ma 2011). The effects of teriparatide on bone mineral density (BMD) are superior to antiresorptive agents at the spine, but its effects at the hip are more modest, and often delayed until the second year of a two-year course of therapy (Leder 2014; Neer 2001).

Parathyroid hormone-related protein (PTHrP; UniProt Accession No. P12272) shares some homology with parathyroid hormone (PTH) at their N-terminal ends, and both proteins bind to the same G-protein coupled receptor, PTH receptor type-1 (PTH1R). Despite a common receptor, PTH primarily acts as an endocrine regulator of calcium homeostasis whereas PTHrP plays a fundamental paracrine role in the mediation of endochondral bone development (Kronenberg 2006). The differential effects of these proteins may be related not only to differential tissue expression, but also to distinct receptor binding properties (Pioszak 2009; Okazaki 2008; Dean 2008). Over the past several years, PTHrP and its secretory forms (PTHrP(1-36), PTHrP(38-94), and osteostatin), as well as analogues thereof, have been investigated as potential treatments for osteoporosis. Subcutaneous injection of PTHrP and its derivatives and analogues has been reported to be effective for treating osteoporosis and/or improving bone healing (Horwitz 2010; Horwitz 2006; Bostrom 2000; Augustine 2013).

Therefore, it is desirable to have an alternative delivery route that is both effective for treatment (e.g., a substantial bioequivalence of the subcutaneous delivery of PTHrP and/or derivatives and analogues thereof) and easy for administration to improve patients' satisfaction and compliance.

SUMMARY OF THE INVENTION

Provided herein in certain embodiments are preparation formulations for use in transdermal delivery of PTHrP analogues such as abaloparatide comprising a PTHrP analogue (e.g., abaloparatide) and one or more excipients selected from the group consisting of Zn²⁺ salts (e.g., ZnCl₂, Zn(OAc)₂, Zn₃(PO₄)₂, ZnCitrate, ZnOxalate, etc., or combinations thereof), Mg²⁺ salts (e.g., MgO, MgCitrate, MgSO₄, MgOrotate, MgLactate, MgCO₃, MgCl₂, Mg(OAc)₂, etc., or combinations thereof) Ca²⁺ salts (e.g., CaSorbate, CaCitrate, CaAscorbate, Ca₃(PO₄)₂, CaCl₂, CaCO₃, CaSO₄, Ca(OAc)₂, etc., or combinations thereof), PEG (polyethylene glycol), PVP (polyvinylpyrrolidone), cyclodextrin (CD, e.g., 2-hydroxypropyl-β-cyclodextrin (HPβCD)), salts of carboxylic acids including fatty acids, NaCl and histidine and various combinations thereof. In certain embodiments, the preparation formulation further comprises water for injection, saline or phosphate buffered saline (PBS). In certain embodiments, the PTHrP analogue comprises, consists of, or consists essentially of abaloparatide ([Glu^22,25, Leu^23,28,31, Aib²⁹, Lys^26,30]hPTHrP(1-34)NH₂), which has the amino acid sequence set forth in SEQ ID NO:1. In certain embodiments, the PTHrP analogue is delivered by a transdermal patch comprising at least one microprojection (e.g., microneedle) prepared using the preparation formulation.

Provided herein in certain embodiments are patches for transdermal administration of a PTHrP analogue comprising one or more microprojections prepared using a preparation formulation as disclosed herein.

Provided herein in certain embodiments are methods of preparing a transdermal patch for administration of a PTHrP analogue comprising preparing at least a microprojection on a blank transdermal patch with a preparation formulation disclosed herein. In certain embodiments, the microprojections are microneedles.

Provided herein in certain embodiments are methods for treating osteoporosis, treating osteopenia, treating osteoarthritis, improving bone mineral density (BMD), improving trabecular bone score (TBS), and treating, preventing, and reducing bone fractures in a subject comprising transdermally administering a therapeutically effective amount of a PTHrP analogue via a transdermal patch comprising at least one microprojections prepared using PTHrP analog preparation formulation as disclosed herein. In some embodiments the osteoporosis being treated is postmenopausal osteoporosis. In some embodiments the osteoporosis being treated is glucocorticoid induced osteoporosis. In certain of these embodiments, the preparation formulation is administered via a transdermal patch as disclosed herein. The bone fractures being treated, prevented, or reduced and the bone with improved BMD or TBS may be vertebral or non-vertebral.

BRIEF DESCRIPTION OF DRAWINGS

This application contains at least one drawing executed in color. Copies of this application with color drawing(s) will be provided by the Office upon request and payment of the necessary fees.”

FIGS. 1A-1C: Pharmacokinetic profile of various formulations of abaloparatide administered by transdermal versus subcutaneous routes. FIG. 1A: One possible bioequivalence “window” of the abaloparatide-SC treatment, % scale on the vertical axis indicates the plasma abaloparatide concentration represented by % of its own maximum (C_max), i.e. 100=C_max, hereinafter referred to as the “normalized plasma concentration.” FIG. 1B: transdermal delivery in monkeys using a preparation formulation of abaloparatide comprising ZnCl₂, the vertical axis indicates normalized peptide plasma concentration. FIG. 1C: transdermal delivery using a preparation formulation of abaloparatide comprising PEG.

FIG. 2: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes (SC). The abaloparatide preparation formulation of the abaloparatide for transdermal delivery did not comprise ZnCl₂or PEG, note the very quick C_maxof the transdermal delivery compare to SC and the increasing pulsatile nature of the delivery. Square: transdermal delivery (TD); and diamond: the abaloparatide-SC treatment. Administration in healthy postmenopausal women, % scale on vertical axis indicates the normalized plasma concentration of abaloparatide represented by % of C_maxfor each group.

FIG. 3: Pharmacokinetic profile of formulations of abaloparatide containing ZnCl₂administered by transdermal (TD) versus subcutaneous (SC) routes, longitude of median plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined. Note the more prolonged release afforded by the ZnCl₂addition.

FIG. 4: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, median plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 5: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, longitude of mean plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 6: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, mean plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 7: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, longitude of median of dose normalized plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 8: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, median of dose normalized plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 9: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, longitude of mean of dose normalized plasma abaloparatide concentration v. time post administration. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 10: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal versus subcutaneous routes, mean of dose normalized plasma abaloparatide concentration v. time post administration in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 11: Comparison of C_maxof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 12: Comparison of C_maxof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 13: Comparison of AUC_lastof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 14: Comparison of AUC_lastof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 15: Comparison of AUC_Infof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 16: Comparison of AUC_Infof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 17: Comparison of C_max/D (C_maxper dosage) of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD combined.

FIG. 18: Comparison of C_max/D (C_maxper dosage) of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 19: Comparison of CL/F of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 20: Comparison of CL/F of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 21: Comparison of HL_Lambda_z of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 22: Comparison of HL_Lambda_z of formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 23: Comparison of T_maxof formulations of abaloparatide administered by transdermal versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 24: Comparison of T_maxof formulations of abaloparatide administered by transdermal (abdomen) versus subcutaneous routes in healthy postmenopausal women. Transdermal administration was to the abdomen with a formulation including ZnCl₂. Historic TD used abaloparatide formulations in PBS buffer (no ZnCl₂) and represents all 150 μg TD studies combined.

FIG. 25: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal route (abdomen) in a selected patient versus historical subcutaneous data in healthy postmenopausal women. TD used abaloparatide formulations PBS buffer with no Zn salt added.

FIG. 26: Percent BMD changes from baseline at lumber spine of the subjects treated with abaloparatide via transdermal delivery or SC injection in postmenopausal women with osteoporosis. Transdermal delivery used abaloparatide formulations with PBS buffer (no Zn salt).

FIG. 27: Percent BMD changes from baseline at total hip of the subjects treated with abaloparatide via transdermal delivery or SC injection in postmenopausal women with osteoporosis. Transdermal delivery used abaloparatide formulations with PBS buffer (no Zn salt).

FIG. 28: Local tolerance data of the subjects treated with abaloparatide via transdermal delivery in postmenopausal women with osteoporosis. Transdermal delivery used abaloparatide formulations with PBS buffer (no Zn salt).

FIG. 29: Pharmacokinetic profile of formulations of abaloparatide administered by transdermal delivery used abaloparatide formulations with PBS only (no Zn salt) in healthy postmenopausal women (diamond) versus subcutaneous (square) routes. Note the very rapid and pulsatile release in the transdermal delivery compared to the SC administration.

FIGS. 30A-30B: PK/PD relationship of formulations of abaloparatide administered by transdermal and by subcutaneous routes all in postmenopausal women with osteoporosis. FIG. 30A: C_maxv. BMD improvement (%) for formulations of abaloparatide administered by transdermal and by subcutaneous routes. FIG. 30B: AUC v. BMD improvement (%) for formulations of abaloparatide administered by transdermal and by subcutaneous routes.

FIG. 31: Comparison of pK curves from a sc combination cohort (80 μg), 1^stgeneration transdermal (200 μg abaloparatide+PBS buffer only) and 2^ndgeneration transdermal (200 μg abaloparatide+ZnCl₂). Values are the geometric means.

FIG. 32: Comparison of pK curves of selected individual patients being treated with the 2^ndgeneration transdermal (200 μg abaloparatide plus ZnCl₂) and compared to a reference set of sc treated patients. Values are the geometric means.

FIG. 33: Concentration-time graph after administration of patch formulated with PEG 3350 NF and 100 μg of abaloparatide to abdomen of healthy post-menopausal women (N=12) where plasma concentration time points are the arithmetic means.

FIG. 34: Concentration-time graph after administration of patch formulated with PEG 3350 NF and 150 μg of abaloparatide to abdomen of healthy post-menopausal women (N=13) where plasma concentration time points are the arithmetic means.

FIG. 35: Concentration-time graph after administration of patch formulated with PEG 3350 NF and 200 μg of abaloparatide to abdomen of healthy post-menopausal women (N=14) where plasma concentration time points are the arithmetic means.

FIG. 36: Concentration-time graph after administration of patch formulated with PEG 3350 NF and ZnCl₂and 100 μg of abaloparatide to abdomen of healthy post-menopausal women (N=8) where plasma concentration time points are the arithmetic means.

FIG. 37: Concentration-time graph after administration of patch formulated with PEG 3350 NF and ZnCl₂and 150 μg of abaloparatide to abdomen of healthy post-menopausal women (N=7) where plasma concentration time points are the arithmetic means.

FIG. 38: Concentration-time graph after administration of patch formulated with PEG 3350 NF and ZnCl₂and 200 μg of abaloparatide to abdomen of healthy post-menopausal women (N=8) where plasma concentration time points are the arithmetic means.

FIG. 39: Transdermal administration utilized patches having different microneedle lengths with abaloparatide formulations (no Zn) in monkeys. Square: subcutaneous delivery; triangle: transdermal delivery (short microneedles—250 μm); diamond: transdermal delivery (regular microneedles—500 μm); and star: transdermal delivery (long microneedles—700 μm).

FIG. 40: Comparison of Cmax (peak plasma concentration (pg/mL)) of various formulations of the PTHrP analogue of SEQ ID NO:1 in monkeys.

FIG. 41: Comparison of AUC (area under the curve) of various formulations of the PTHrP analogue of SEQ ID NO:1 administered by transdermal versus subcutaneous route.

FIG. 42: Comparison of plasma concentration (pg/mL) of formulation of abaloparatide (ABL, SEQ ID NO:1) administered by subcutaneous route (SC) or transdermal administration (TD), wherein the transdermal administration utilized a transdermal patch coated using different transdermal formulations in monkeys.

Table 1. Modeling of TD-A32 data for bioequivalence for the abaloparatide-SC treatment.

Table 2. PK Results of Abaloparatide 100 μg TD, Abaloparatide 150 μg TD, Abaloparatide 200 μg TD, Abaloparatide 80 μg SC, and Historical 150 μg TD.

Table 3. Comparisons of Abaloparatide 100 μg TD, Abaloparatide 150 μg TD, Abaloparatide 200 μg TD, Abaloparatide 80 μg SC, and Historical 150 μg TD to UnoPen 80 μg SC, respectively.

Table 4. Comparisons of Abaloparatide 100 μg TD, Abaloparatide 150 μg TD, Abaloparatide 200 μg TD, and Abaloparatide 80 μg SC to Historical 150 μg TD, respectively.

Table 5. Design for a Phase 2 study of transdermal delivery of abaloparatide using a transdermal patch prepared by a first general abaloparatide formulation (with PBS).

Table 6. C_max, AUC, and BMD improvement of a Phase 2 study of transdermal delivery of abaloparatide (TD-50 mcg, TD-100 mcg, and TD-150 mcg) using a transdermal patch prepared by a first general abaloparatide formulation (with PBS), and subcutaneous delivery of abaloparatide (SC-80 mcg).

DETAILED DESCRIPTION

Abaloparatide is a synthetic PTHrP analogue having the sequence set forth in SEQ ID NO: 1. (Ala-Val-Ser-Glu-His-Gln-Leu-Leu-His-Asp-Lys-Gly-Lys-Ser-Ile-Gln-Asp-Leu-Arg-Arg-Arg-Glu-Leu-Leu-Glu-Lys-Leu-Leu-Aib-Lys-Leu-His-Thr-Ala, SEQ ID NO:1), Aib is 2-aminoisobutyric acid, also known as α-aminoisobutyric acid. Abaloparatide has shown potent anabolic activity with decreased bone resorption, less calcium-mobilizing potential, and improved room temperature stability (Obaidi 2010). Studies performed in animals have demonstrated marked bone anabolic activity following administration of abaloparatide, with complete reversal of bone loss in ovariectomy-induced osteopenic rats and monkeys (Doyle 2013a; Doyle 2013b; Hattersley 2013). Abaloparatide has been developed as a promising anabolic agent for the treatment of osteopenia (e.g., glucocorticoid-induced osteopenia), osteoporosis (e.g. glucocorticoid-induced osteoporosis), and/or osteoarthritis.

Subcutaneous administration of 80 μg abaloparatide (hereinafter the “abaloparatide-SC treatment”) has been shown to significantly reduce incidences of new vertebral, non-vertebral, major osteoporotic and clinical fractures versus a placebo. Subcutaneous abaloparatide administration has also been shown to improve bone mineral density (BMD) and/or trabecular bone score (TBS) of treated subjects at the lumbar spine, total hip, and femoral neck. In certain embodiments, the abaloparatide-SC treatment comprises subcutaneous administration of an aqueous formulation comprising abaloparatide (about 2 mg/mL) in an acetate buffer, with a pH of about 4.5 to about 5.6, or about 5.1. Optionally, the aqueous formulation further comprises phenol (about 5 mg/mL). In certain examples of these embodiments, the acetate buffer comprises tri-hydrate sodium acetate (about 5 mg/mL) with pH (e.g., about 4.5 to about 5.6, or about 5.1) adjusted with acetic acid.

Transdermal administration of abaloparatide is an attractive alternative to subcutaneous administration due to its less invasive nature. However, transdermal administration may have different PK profile compared to subcutaneous administration. It has been found that AUC of transdermal abaloparatide administration and subcutaneous abaloparatide administration (80 μg) had a linear relationship with the achieved BMD changes from the baseline after 6 months of treatments (FIG. 30B). It is thus desired to develop transdermal abaloparatide administrations that are substantially bioequivalent to the subcutaneous abaloparatide administration to benefit from both the preferred osteoanabolic profile of subcutaneous abaloparatide administration and the convenience of transdermal administration.

As disclosed herein, it has been unexpectedly found that transdermal abaloparatide administration using a patch prepared with a preparation formulation comprising abaloparatide and one or more excipients selected from the group consisting of ZnCl₂, PEG, and histidine produces a substantial bioequivalence to subcutaneous administration in a monkey model. Furthermore, preliminary clinical studies indicate that ZnCl₂in a transdermal formulation blunts the pulsatile nature compared with a non-ZnCl₂containing formulation and pushes the curve into one resembling the sc curve. This is a notable achievement in that achieving bioequivalence to a sc dose of abaloparatide would indicate exceptional fracture prevention effects as has been reported for 80 μg sc administration.

Based on these findings, provided herein are PTHrP analogue preparation formulations, transdermal patches prepared using these preparation formulations, transdermal patches comprising these preparation formulations, methods of making these patches, and methods of using the disclosed preparation formulations and patches to administer PTHrP analogues in a transdermal manner and to treat osteoporosis, osteopenia, and osteoarthritis, improve BMD, improve TBS, and treat, prevent, and reduce bone fractures in a subject. In certain embodiments of the preparation formulations, transdermal patches, and methods provided herein, the PTHrP analogue is abaloparatide consisting of the amino acid sequence set forth in SEQ ID NO:1, or an abaloparatide derivative comprising or consisting essentially of the amino acid sequence set forth in SEQ ID NO:1. In certain embodiments of the preparation formulations, transdermal patches, and methods provided herein, the transdermal delivery of the PTHrP analogue produces substantial bioequivalence or bioequivalence to a subcutaneous delivery of abaloparatide at the dosage of about 20 μg to about 200 μg, about 40 μg to about 120 μg, about 60 μg to about 100 μg, about 70 μg to about 90 μg, or about 80 μg. In certain embodiments of the preparation formulations, transdermal patches, and methods provided herein, the transdermal delivery of the PTHrP is a substantial bioequivalence or bioequivalence of the abaloparatide-SC treatment.

As used herein, two treatments of an active agent are bioequivalent to one another if the 90% confidence interval of the ratio of area under the curve (AUC) and/or the peak serum concentration of the active agent (C_max) falls completely within the range 80-125%. See, e.g., FIG. 1A showing a bioequivalence window of the abaloparatide-SC treatment in Chinese Cynomolgus monkeys. Serum abaloparatide concentrations are presented as percentage of C_max.

As used herein, the term “substantially,” “substantial” or “essentially” means nearly completely or completely. In particular, a normal bioequivalent range means a compound in a particular formulation for transdermal delivery within the 80%-125% (for the mean in the 90% confidence interval (CI)) of AUC (_0-t, _0-inf) and C_maxof the reference compound in a reference formulation. In some embodiments the reference formulation is the SC delivery of 80 μg abaloparatide formulated as described herein. In certain embodiments, the transdermal delivery of a compound or more particularly abaloparatide falls within a substantially bioequivalent range wherein said range is 70%-136%, or 65%-141%, or 60%-147%, or 50%-158% (for the mean in the 90% confidence interval (CI)) of AUC (_0-t, _0-inf) and C_maxof the reference compound in a reference formulation.

As used herein, the term “about” or “approximately” means a range of ±0-10%, or ±0-5% or the numeral following the term.

As used herein, the term “transdermal delivery” refers to a delivery of an active agent through the stratum corneum to make contact with the intradermal space without significant pain upon penetration. Because the stratum corneum has no nerves, it may be pierced without stimulating nerves. The terms “transdermal” and “intradermal” are used interchangeably herein. The stratum corneum is composed primarily of several layers of dead skin cells and is not vascularized. Thus, the stratum corneum often poses a formidable barrier to the transdermal delivery of an active agent, especially for charged macromolecules such as peptides. Unlike active agents delivered by subcutaneous injection, which almost provides a complete entrance into the blood stream, many factors (and barriers) can affect the pharmacokinetics of drugs delivered by a transdermal route. For example, the site of application, the thickness, integrity, and hydration condition of the skin, the thickness and density of the adipose tissue under the skin of the application site, the size of the drug molecules, the pH condition and permeability of the membrane of the transdermal device, etc., all may affect the bioavailability of drugs delivered transdermally. In certain embodiments, transdermal delivery involves penetrating the skin through the stratum corneum into the dermis to a depth of up to about 700 μm, or up to about 600 μm, or up to about 500 μm, or up to about 400 μm, or up to about 300 μm, or up to about 250 μm, or up to about 150 μm. In some embodiments, the average needle depth of penetration is approximately 800 μm, or about 700 μm, or about 600 μm, or about 500 μm, or about 400 μm, or about 300 μm, or about 250 μm, or about 150 μm.

I. Preparation Formulation for Transdermal Delivery

Provided herein in certain embodiments are preparation formulations for transdermal delivery of a therapeutically active substance, e.g., a bioactive peptide, a bioactive peptide contains at least 10 amino acids, e.g., a PTHrP analogue (e.g., comprising, consisting essentially of, or consisting of abaloparatide). In certain embodiments, the transdermal delivery produces a substantial bioequivalence or bioequivalence to subcutaneous delivery of the PTHrP analogue (e.g., at 80 μg). These formulations comprise a PTHrP analogue and one or more excipients selected from the group consisting of salts of Zn²⁺, salts of Mg²⁺, salts of Ca²⁺, salts of histidine, salts of carboxylic acids (e.g., fatty acids), NaCl, PEG, PVP, cyclodextrin (CD, e.g., 2-hydroxypropyl-β-cyclodextrin (HPβCD)), and combinations thereof. In certain embodiments the salt of Zn²⁺ is selected from the group consisting of Zn(OAc)₂, ZnCl₂, Zn₃(PO₄)₂, zinc citrate (ZnCitrate), zinc oxalate (ZnOxalate), and combinations thereof, the salt of Ca²⁺ is selected from the group consisting of calcium sorbate (CaSorbate), calcium citrate (CaCitrate), calcium ascorbate (CaAscorbate), Ca₃(PO₄)₂, CaCl₂, CaCO₃, CaSO₄, Ca(OAc)₂and combinations thereof, the salt of Mg²⁺ is selected from the group consisting of MgO, magnesium citrate (MgCitrate), MgSO₄, magnesium orotate (MgOrotate), magnesium lactate (MgLactate), MgCO₃, MgCl₂, Mg(OAc)₂, and combinations thereof. In certain embodiments, two or more salts of Mg²⁺, Zn²⁺ and/or Ca²⁺ as described herein are combined together for purposes of a transdermal formulation. In certain embodiments, the preparation formulation further comprises water for injection, brine or PBS. In certain embodiments, the PTHrP analogue comprises, consists of, or consists essentially of abaloparatide. In certain embodiments, the transdermal delivery of the PTHrP analogue produces substantial bioequivalence or bioequivalence to a subcutaneous delivery of abaloparatide at the dosage of about 20 μg to about 200 μg, about 40 μg to about 120 μg, about 60 μg to about 100 μg, about 70 μg to about 90 μg, or about 80 μg. In certain embodiments, the transdermal delivery of the PTHrP is a substantial bioequivalence or bioequivalence of the abaloparatide-SC treatment. In certain embodiments, the PTHrP analogue is delivered by a transdermal patch comprising at least one microprojection (e.g., microneedle) prepared using the preparation formulation.

In certain embodiments, the preparation formulation comprises PEG with a molecular weight of about 3,000 to about 3,700, about 2,000 to about 5,000, about 3,00 to about 3,500, or about 1,000 to about 6,000. A concentration by weight of PEG to the total amount of the preparation formulation is about 0.01% to about 50%, about 5% to about 50%, about 5% to about 45%, about 5% to about 40%, about 5% to about 35%, about 5% to about 30%, about 5% to about 25%, about 5% to about 20%, about 5% to about 15%, about 10% to about 50%, about 10% to about 45%, about 10% to about 40%, about 10% to about 35%, about 10% to about 30%, about 10% to about 25%, about 10% to about 20%, about 10% to about 15%, about 15% to about 50%, about 15% to about 45%, about 15% to about 40%, about 15% to about 35%, about 15% to about 30%, about 15% to about 25%, about 15% to about 20%, about 13% to about 17%, about 14% to about 16%, or about 14.9%.

In certain embodiments, the preparation formulation comprises water and a Zn²⁺ salt (also referred to as Zn salt, salt of Zn, or salt of Zn₂±), in some embodiments said coating formulation comprises ZnCl₂, or Zn(OAc)₂, or Zn₃(PO₄)₂, or ZnCitrate or ZnOxalate or combinations thereof. The concentration of Zn²⁺ salt (e.g., ZnCl₂) in the preparation formulation, for example, by weight to the total amount of the preparation formulation is about 0.01% to about 30%, 0.1% to about 30%, 0.3% to about 30%, about 0.5% to about 30%, about 0.8% to about 30%, about 1% to about 30%, about 1.5% to about 30%, about 2% to about 30%, about 5% to about 30%, 10% to about 30%, 15% to about 30%, about 20% to about 30%, about 25% to about 30%, about 0.01% to about 20%, 0.1% to about 20%, 0.3% to about 20%, about 0.5% to about 20%, about 0.8% to about 20%, about 1% to about 20%, about 1.5% to about 20%, about 2% to about 20%, about 5% to about 20%, 10% to about 20%, 15% to about 20%, about 0.01% to about 10%, 0.1% to about 10%, 0.3% to about 10%, about 0.5% to about 10%, about 0.8% to about 10%, about 1% to about 10%, about 1.5% to about 10%, about 2% to about 10%, about 5% to about 10%, about 0.01% to about 5%, 0.1% to about 5%, 0.3% to about 5%, about 0.5% to about 5%, about 0.8% to about 5%, about 1% to about 5%, about 1.5% to about 5%, about 2% to about 5%, about 0.01% to about 3%, 0.1% to about 3%, 0.3% to about 3%, about 0.5% to about 3%, about 0.8% to about 3%, about 1% to about 3%, about 1.5% to about 3%, about 2% to about 3%, about 0.01% to about 30%, 0.1% to about 30%, 0.3% to about 30%, about 0.5% to about 30%, about 0.8% to about 2%, about 1% to about 2%, about 1.5% to about 2%, about 0.01% to about 1%, 0.1% to about 1%, 0.3% to about 1%, about 0.5% to about 1%, about 0.8% to about 1%, or about 0.8%. In certain embodiments, the coating formulation for the described ranges comprises a Ca²⁺ salt wherein said Ca²⁺ salt can include CaSorbate, CaCitrate, CaAscorbate, Ca₃(PO₄)₂, CaCl₂, CaCO₃, CaSO₄, Ca(OAc)₂or combinations thereof. Ca²⁺ salt is also referred to as Ca salt, salt of Ca, or salt of Ca²⁺. In some embodiments the coating solution comprises a Mg²⁺ salt wherein said Mg²⁺ salt can include MgO, MgCitrate, MgSO₄, MgOrotate, MgLactate, MgCO₃, MgCl₂, Mg(OAc)₂, or combinations thereof. Mg²⁺ salt is also referred to as Mg salt, salt of Mg, or salt of Mg²⁺.

In some embodiments of this invention, a formulated patch ready to package and use (the initial coating solution dried to remove water) is provided wherein said formulated patch comprises a Zn²⁺ salt. In certain embodiments the Zn²⁺ salt is Zn(OAc)₂, in some embodiments the Zn²⁺ salt is ZnCl₂, in certain embodiments it is Zn₃(PO₄)₂, in some embodiments it is ZnCitrate and in certain embodiments it is ZnOxalate or combinations thereof. In certain embodiments, the formulated patch is made by coating with the coating solution in one or multiple coating iterations and then drying said patch or allowing said patch to dry to a fairly constant weight and then characterizing said patch as weight by percent of the salt, the metal including its counterions. In certain embodiments, the coated patch, dried and ready to use comprises from 1.0 to 20% Zn salt by weight. In certain embodiments, said coated patch comprises from 1.5% to 15% Zn salt by weight. In some embodiments, the coated and dried patch comprises 1.5% to 10% Zn salt by weight, or 1.8%-8.5%, or 1.9% to 5.9%, or about 1.9% to 8.5%, or about 2.0% to about 8%, or 5% to 8% by weight or is between 1.7% to 2.25%, or between 5 to 7%, or about 5.8%, or about 1.9%.

In some embodiments of this invention, a formulated patch ready to package and use (the initial coating solution dried to remove water) is provided wherein said formulated patch comprises a Ca²⁺ salt. In some embodiments the Ca²⁺ salt is CaSorbate, CaCitrate, CaAscorbate, Ca₃(PO₄)₂, CaCl₂, CaCO₃, CaSO₄, Ca(OAc)₂or combinations thereof. In certain embodiments, the formulated patch is made by coating with the coating solution in one or multiple coating iterations and then drying said patch or allowing said patch to dry to a fairly constant weight and then characterizing said patch as weight by percent of the salt, the metal including its counterions. In certain embodiments, the coated patch, dried and ready to use comprises from 1.0 to 20% Ca salt by weight. In certain embodiments, said coated patch comprises from 1.5% to 15% Ca salt by weight. In some embodiments, the coated and dried patch comprises 1.5% to 10% Ca salt by weight, or 1.8%-8.5%, or 1.9% to 5.9%, or about 1.9% to 8.5%, or about 2.0% to about 8%, or 5% to 8% by weight.

In some embodiments of this invention, a formulated patch ready to package and use (the initial coating solution dried to remove water) is provided wherein said formulated patch comprises a Mg²⁺ salt. In some embodiments the Mg²⁺ salt is MgO, MgCitrate, MgSO₄, MgOrotate, MgLactate, MgCO₃, MgCl₂, Mg(OAc)₂, or combinations thereof. In certain embodiments, the formulated patch is made by coating with the coating solution in one or multiple coating iterations and then drying said patch or allowing said patch to dry to a fairly constant weight and then characterizing said patch as weight by percent of the salt, the metal including its counterions. In certain embodiments, the coated patch, dried and ready to use comprises from 0.5 to 15% Mg salt by weight. In certain embodiments, said coated patch comprises from 1.0% to 10% Mg salt by weight. In some embodiments, the coated and dried patch comprises 1.5% to 10% Mg salt by weight, or 1.8%-8.5%, or 1.9% to 5.9%, or about 1.9% to 8.5%, or about 2.0% to about 8%, or 5% to 8% by weight.

In certain embodiments, the formulated patch comprises two or more of the Zn²⁺, Ca²⁺ and/or Mg²⁺ salts described immediately above.

Example of Coating Solution

Component
Function
Weight %

Abaloparatide
API
35.78

Zinc Chloride, USP
Complexing Agent
2.22

(ZnCl₂)

Sterile Water for
Solvent*
62.00

Injection, USP

Total
100