Stabilized modified-release vitamin d formulations and methods of administering the same

Abstract

The present invention relates to stabilized modified release vitamin D formulations and methods of administering the same. A stabilized formulation for controlled release of a vitamin D compound is disclosed. The formulation comprises one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 and a cellulose compound. The stabilized formulation exhibits a stable dissolution profile after exposure to storage conditions and demonstrates improved pharmacokinetic parameters compared to unstabilized formulations.

Description

[0001] Divisional Statement

[0002] This application is a divisional application of the application patent application with the application number 201480024061.X, filed on March 14, 2014, and the title "Stabilized modified release vitamin D formulations and methods of administering the same".

[0003] Cross Reference to Related Applications

[0004] This claims the benefit under 35 U.S.C. § 119(e) of U.S. Provisional Patent Application Serial No. 61 / 801,896, filed March 15, 2013, and the disclosure of which is incorporated herein by reference. TECHNICAL FIELD

[0005] The present disclosure relates generally to controlled release pharmaceutical compositions. More particularly, the present invention relates to controlled release formulations for delivering vitamin D compounds, such as 25-hydroxyvitamin D compounds, for intestinal absorption, which are storage stable over time. BACKGROUND

[0006] The vitamin D metabolites known as 25-hydroxyvitamin D2and 25-hydroxyvitamin D3(collectively "25-hydroxyvitamin D") are fat-soluble prohormones that help maintain normal levels of calcium and phosphorus in the blood stream. The prohormones 25-hydroxyvitamin D2are produced from vitamin D2(ergocalciferol) and 25-hydroxyvitamin D3from vitamin D3(cholecalciferol), primarily by one or more enzymes located in the liver. These two prohormones can also be produced extrarenally from vitamin D2and vitamin D3(collectively "vitamin D") in certain cells, such as intestinal epithelial cells, that contain the same or similar enzymes as those found in the liver.

[0007] The 25-hydroxyvitamin D prohormones are further metabolized in the kidney to the active vitamin D hormones. The prohormone 25-hydroxyvitamin D2is metabolized to lα,25-dihydroxyvitamin D2; likewise, 25-hydroxyvitamin D3is metabolized to lα,25-dihydroxyvitamin D3(calcitriol). The production of these active hormones from the 25-hydroxyvitamin D prohormones can also occur extrarenally in cells that contain the required enzymes.

[0008] Controlled release formulations of 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3can be administered to treat 25-hydroxyvitamin D insufficiency and deficiency without the physiologically excessive surges in intraluminal, intracellular, and blood levels of 25-hydroxyvitamin D and their consequences; without causing substantially increased catabolism of the administered 25-hydroxyvitamin D; and without causing serious side effects associated with vitamin D supplementation, i.e., vitamin D toxicity. The controlled release formulations are effective in reducing PTH levels without undesirable increases in serum calcium and serum phosphorus, and are thus useful in treating secondary hyperparathyroidism, e.g., in CKD patients. See International Patent Application Nos. PCT / US2007 / 061521 and PCT / US2008 / 061579 and U.S. Patent Application No. 12 / 109,983, incorporated herein by reference.

[0009] Controlled release compositions provide substantially increased absorption of 25-hydroxyvitamin D via transport on DBP and reduced absorption via transport in chylomicrons. These compositions also provide maintenance of substantially constant blood levels of 25-hydroxyvitamin D during the post-dosing period of 24 hours. By providing gradual, sustained, and direct release of 25-hydroxyvitamin D2 / 25-hydroxyvitamin D3and preferential absorption to circulating DBP (rather than chylomicrons), blood, intraluminal, and intracellular 25-hydroxyvitamin D concentration spikes, i.e., supraphysiological levels and associated inappropriate catabolism, can be reduced or eliminated. Furthermore, by providing gradual and sustained release, serum levels of 25-hydroxyvitamin D can be increased and maintained in a more predictable manner than administration of immediate release formulations, allowing consistent dosing and reducing or eliminating the need for frequent patient monitoring.

[0010] To deliver the benefits of controlled release formulations of 25-hydroxyvitamin D to patients, there is a need for stabilized pharmaceutical compositions that retain the desired dissolution characteristics of the formulations for extended periods of time, e.g., after shipping and storage. SUMMARY

[0011] The present invention comprises a controlled release vitamin D formulation comprising a vitamin D compound and a cellulose compound.

[0012] The present invention also encompasses a storage-stabilized formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting such formulation. In one aspect, the stabilized formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3and a stabilizing agent or stabilizing compound, e.g., a cellulose compound. The stabilized formulations of the present invention with the recited stabilizing agents can have improved or relatively improved "storage stability" or post-aging stability, as well as one or more additional features, including improved physical, chemical, and biological properties, compared to the disclosed formulations without such agents. The claimed formulations are thus suitable as therapeutic agents with long shelf life and improved bioavailability compared to aged, unstable formulations.

[0013] In one embodiment, the stabilized formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a lipophilic matrix (e.g., a wax matrix), and a stabilizing agent (e.g., a cellulose compound). In one aspect, the stabilized formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a wax matrix, and a cellulose stabilizing agent. In another aspect, the formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a wax matrix, and an effective amount of a cellulose compound to maintain the advantageous degree of stabilization described herein.

[0014] In one type of embodiment, the stabilized formulation comprises a mixture of an active agent-loaded wax matrix comprising one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3and a cellulose stabilizing agent, wherein the formulation releases an amount of 25-hydroxyvitamin D during in vitro dissolution that varies by 30% or less at all dissolution test time points compared to the amount released at the same dissolution time points during in vitro dissolution of a fresh product after exposure to storage conditions of 25°C and 60% relative humidity for at least one month.

[0015] In one type of embodiment, the formulation is an improved formulation for controlled release of a vitamin D compound. In one aspect, the improvement comprises admixing a stabilizing agent into a formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation. In another aspect, the improvement comprises admixing an effective amount of a cellulose compound into a formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation to provide the advantageous degree of stability described herein.

[0016] In one embodiment, the present application comprises a stable, sustained release vitamin D formulation comprising 25-hydroxyvitamin D2or 25-hydroxyvitamin D3or a combination thereof and a sustained release excipient, wherein the formulation has a dissolution profile X at TO, which maintains this profile under storage conditions selected from, for example, room temperature and ambient humidity, or 25°C and 60% RH, or 40°C and 75% RH, according to the formula X = TO + / - 30%.

[0017] Other aspects and advantages will be apparent to those of ordinary skill in the art from the review of the specification in conjunction with the drawings. While the composition and methods are susceptible to various modifications and alternative forms, specific embodiments are shown by way of example in the drawings and the description herein, which is not intended to limit the disclosure to the particular embodiments described. It should be understood that both the drawings and the description are illustrative and are not to be construed as placing limitations upon the scope of the disclosure.

[0018] Optional features, including, but not limited to, components, ranges of constituents thereof, alternatives, conditions, and steps, for the compositions and methods described herein are contemplated to be selected from the various aspects, embodiments, and examples provided herein. BRIEF DESCRIPTION OF DRAWINGS

[0019] FIG. 1 shows the dissolution profile of formulations according to the present disclosure after storage at 25°C and 60% relative humidity for 0 to 24 months. Dissolution time (hours) is plotted on the x-axis and the average percent of 25-hydroxyvitamin D3dissolved is shown on the y-axis. Figure 1A , 1B and 1C show the dissolution profile of formulations comprising 30 pg, 60 pg, and 90 pg of 25-hydroxyvitamin D3, respectively.

[0020] FIG. 2 shows the dissolution profile of formulations according to the present disclosure after storage at 40°C and 75% relative humidity for 0 to 6 months. Dissolution time (hours) is plotted on the x-axis and the average percent of 25-hydroxyvitamin D3dissolved is shown on the y-axis. Figure 2A , 2B and 2C show the dissolution profile of formulations comprising 30 pg, 60 pg, and 90 pg of 25-hydroxyvitamin D3, respectively.

[0021] Figure 3 shows the dissolution profile of formulations after storage at 25°C and 60% relative humidity for 0 to 12 months. Dissolution time (hours) is plotted on the x-axis and the average labeled % of 25-hydroxyvitamin D3released is shown on the y-axis. Figure 3 A shows the dissolution profile of comparative formulations without a cellulose compound. Figure 3 B shows the dissolution profile of stabilized formulations according to the present disclosure.

[0022] Figure 4The resulting mean baseline-adjusted calcifediol concentration for the treatment group (PK population) of patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0023] Figure 5 The resulting summary baseline-adjusted PK parameters for the calcifediol concentration for the treatment group (PK population) of patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0024] Figure 6 The resulting mean baseline-adjusted serum 1,25-dihydroxyvitamin D level during the 6-week treatment (PK population) for the patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0025] Figure 7 The resulting summary baseline-adjusted repeat-dose PK parameters for serum 1,25-dihydroxyvitamin D for the treatment group (PK population) of patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0026] Figure 8 The resulting mean baseline percent change in plasma iPTH level during the 6-week treatment (PK population) for the patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0027] Figure 9 The resulting summary baseline-adjusted repeat-dose PK parameters for plasma iPTH for the treatment group (PK population) of patients described in Example 4 treated with a formulation according to the present disclosure is shown.

[0028] Figure 10 and 11 The resulting percent change from baseline in plasma iPTH versus baseline-adjusted calcifediol and 1,25-dihydroxyvitamin D exposure (AUC 0-6周 ) at EOT for the PK population of patients described in Example 4 treated with a formulation according to the present disclosure is shown. DETAILED DESCRIPTION

[0029] The terms "controlled release" and "modified release" as used herein are used interchangeably and refer to the administered vitamin D compound being released in a manner that deviates from immediate release. The terms "sustained release" and "extended release" as used herein are used interchangeably and refer to the administered vitamin D compound being released over a longer period of time compared to a comparable immediate release formulation, such that the serum concentration of the vitamin D compound remains elevated above baseline for a longer period of time compared to a comparable immediate release formulation. The foregoing terms optionally include a delayed release feature. For example, a controlled release formulation would feature a delayed release type of C max greater than that of an immediate release formulation at a certain time maxAs another example, release of the 25-hydroxyvitamin D compound will preferably be at a rate that maintains or elevates the total serum or blood level of 25-hydroxyvitamin D above pre-dosing levels for a prolonged period of time, e.g., 4 to 24 hours or even longer.

[0030] Unless otherwise specified, the term "cellulose compound" as used herein can include cellulose (C6H 10 O5) n or a cellulose derivative. A "cellulose ether" is a cellulose derivative that has been chemically modified to etherize some or all of the hydroxyl moieties in the cellulose molecule. Examples of cellulose derivatives that can be used as stabilizing agents include, but are not limited to, celluloronic acid, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, polyanionic cellulose, and combinations thereof, by way of example. Each cellulose compound or stabilizing agent is also encompassed by these terms corresponding to different grades of, e.g., molecular weight, viscosity, solubility, and hydration.

[0031] Any vitamin D compound and combinations thereof suitable for prophylactic and / or therapeutic use are contemplated for inclusion in the formulations described herein. Vitamin D, 25-hydroxyvitamin D, 1,25-dihydroxyvitamin D, and other vitamin D metabolites and analogs are also suitable for use as active compounds in pharmaceutical compositions. Particular examples include, but are not limited to, vitamin D3 (cholecalciferol), vitamin D2 (ergocalciferol), 25-hydroxyvitamin D3, 25-hydroxyvitamin D2, 25-hydroxyvitamin D4, 25-hydroxyvitamin D5, 25-hydroxyvitamin D7, 1a,25-dihydroxyvitamin D3, 1a,25-dihydroxyvitamin D2, 1a,25-dihydroxyvitamin D4, and vitamin D analogs (including all hydroxy and dihydroxy forms), including 1,25-dihydroxy-19-nor- vitamin D2and 1a-hydroxyvitamin D3. In one type of embodiment, the vitamin D compound includes one or more hydroxy forms, such as a combination of 25-hydroxyvitamin D3and 25-hydroxyvitamin D2.

[0032] The type of vitamin D compound specifically contemplated for use in the formulations disclosed herein can include 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, or combinations thereof. 25-hydroxyvitamin D3is specifically contemplated. The term 25-hydroxyvitamin D as used herein refers to one or more of 25-hydroxyvitamin D3, 25-hydroxyvitamin D2, 25-hydroxyvitamin D4, 25-hydroxyvitamin D5, or 25-hydroxyvitamin D7, and it is contemplated that the preferred embodiments in any reference to this are one or more of 25-hydroxyvitamin D3and 25-hydroxyvitamin D2, preferably 25-hydroxyvitamin D3. Thus, in any and all of the formulations described herein, it is specifically contemplated that the active agent can include one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, particularly 25-hydroxyvitamin D3. In the disclosure herein, the vitamin D compound (or combinations thereof) is also referred to as the "active" portion (or "active" agent) of the formulation, as distinguished from the controlled release matrix, stabilizing agents, and other excipients. In the pharmacokinetic testing reported herein with respect to samples using 25-hydroxyvitamin D3as the active agent, references to 25-hydroxyvitamin D should be interpreted to mean 25-hydroxyvitamin D3, and all pharmacokinetic (PK) results related to (e.g., t max , C max , AUC) should be understood to be based on 25-hydroxyvitamin D3.

[0033] A "stabilized" formulation as used herein refers to a formulation that exhibits a stable in vitro dissolution profile according to any of the parameters further described herein and controlled release (e.g., sustained release) of the vitamin D compound in vivo for a period of time after initial manufacture, e.g., after actual shelf storage or accelerated stability storage conditions. Release of the active ingredient can be measured using a suitable in vitro dissolution method, such as one of the methods known in the art. In principle, any of the dissolution studies described in the United States Pharmacopeia, USP 29-NF 24, Dissolution <711> Physical Tests and Determinations, United States Pharmacopeial Convention, Inc., Rockville, MD, 2006, pp. 2673-2682; European Pharmacopoeia 2.9.3 Dissolution Test for Solid Dosage Forms; or Japanese Pharmacopoeia 6.10 Dissolution Test can be used to determine whether a formulation is stable. For purposes of the present invention, the in vitro dissolution method is United States Pharmacopeia, USP 29-NF 24, Dissolution <711> Physical Tests and Determinations, United States Pharmacopeial Convention, Inc., Rockville, MD, 2006, pp. 2673-2682, using Apparatus 2 (paddle method), as described in the Examples below.

[0034] As used herein, t max The time (or Tmax) is defined as the time at which the plasma concentration of the active compound reaches its maximum within the dosing interval after administration of a formulation according to the present invention. When a single 25-hydroxyvitamin D compound, e.g., 25-hydroxyvitamin D3, is administered, t max The time is defined as the time at which the plasma concentration of serum 25-hydroxyvitamin D3reaches its maximum within the dosing interval after administration of a formulation.

[0035] In accordance with the NKF K / DOQI guidelines, as used herein, vitamin D sufficiency is defined as serum 25-hydroxyvitamin D levels > 30 ng / mL, vitamin D insufficiency is defined as serum 25-hydroxyvitamin D of 16-30 ng / mL, mild vitamin D deficiency is defined as serum 25-hydroxyvitamin D of 5-15 ng / mL, and severe vitamin D deficiency is defined as serum 25-hydroxyvitamin D less than 5 ng / mL.

[0036] In jurisdictions where methods of treatment of the human body are not patentable, the meaning of "administering" a composition to a human subject should be limited to prescribing a regulated substance to a human subject who will self-administer by any technique (e.g., orally, inhalation, topical application, injection, insertion, etc.). The broadest reasonable interpretation consistent with the law or regulation defining patentable subject matter is intended. In jurisdictions where methods of treatment of the human body are patentable, "administering" of a composition includes methods of treatment of the human body as well as the foregoing activities.

[0037] It is specifically intended that any numerical values recited herein include all values from the lower to the upper end of the range, i.e., each numerical value is intended to include all possible combinations of the numerical values recited as the lower and upper limits of the range. For example, if a concentration range or a beneficial effect range is stated as 1% to 50%, it is intended that values such as 2% to 40%, 10% to 30%, or 1% to 3%, etc., are expressly enumerated in this specification. As another example, a stated concentration of about 20% is intended to encompass values from 19.5% to 20.5%. These are only specific examples of what is intended in this regard.

[0038] Disclosed herein are formulations for controlled release of a vitamin D compound in the gastrointestinal tract of a subject who ingests the formulation. Such formulations will include a vitamin D compound as described herein, a matrix component (e.g., a lipophilic matrix) that releasably binds the vitamin D compound and controllably releases the vitamin D compound, and a stabilizing agent (e.g., a cellulosic compound).

[0039] A stabilized formulation according to the disclosure herein releases an amount of 25-hydroxyvitamin D in an in vitro dissolution that is not substantially different from the same formulation immediately after manufacture and prior to storage, after being stored for a period of time. For example, in one embodiment, a formulation releases an amount of 25-hydroxyvitamin D in an in vitro dissolution that varies by 30% or less at any given dissolution time point after four hours, after being stored for two months at storage conditions of 25°C and 60% relative humidity, as compared to the amount released at the same dissolution time point during an in vitro dissolution performed prior to exposing the formulation to the storage conditions (i.e., a freshly produced product).

[0040] The table below provides examples of the degree of advantageous storage stability expected for embodiments of the application after storage at 25 °C and 60% RH and alternatively at 40 °C and 75% RH for various times after initial manufacture and at various times during the dissolution test. The degree of storage stability is expressed in terms of the maximum deviation from nominal active potency, i.e., the maximum change % from LC. Alternative embodiments of the maximum deviation are also provided.

[0041]

[0042]

[0043] In one type of embodiment, the formulation will have the degree of advantageous stability described in the table just above at multiple time points throughout the dissolution test, e.g., at least at the 2 hour and 4 hour time points, optionally yet at the 6 hour time point, further optionally yet at the 8 hour time point, and further optionally yet at the 12 hour time point, such that the dissolution profile after storage follows that of the fresh product. Alternatively, the formulation will have the degree of advantageous stability described in the table just above at least at the 2, 6, and 12 hour time points. Alternatively, the formulation will have the degree of advantageous stability described in the table just above at least at the 4, 8, and 12 hour time points. Alternatively, the formulation will have the degree of advantageous stability described in the table just above at least at the 2, 4, and 6 hour time points. Alternatively, the formulation will have the degree of advantageous stability described in the table just above at least at the 4, 6, 8, and 12 hour time points, or at the 4 hour and all times thereafter.

[0044] In any and all embodiments described in the table just above, the expected deviation can be positive (more release) or negative (less release) with respect to the fresh product. In one type of embodiment, the expected deviation will be in the negative (less release) direction at multiple time points. Further, in one type of embodiment, the expected deviation in dissolution release will be negative (less release) at multiple time points if there is no presence of a stabilizing agent in the formulation.

[0045] In any embodiment contemplated herein, the dissolution release profile of the formulation can have the characteristics of any of the examples provided herein below. For example, the dissolution release profile of the formulation can be one that provides the following release of the vitamin D compound: less than 30% at 2 hours, greater than 45% at 6 hours, and greater than 80% at 12 hours, and further optionally less than 60% at 6 hours.

[0046] In another type of embodiment, the formulation can be characterized by an in vitro dissolution profile providing release of the vitamin D compound of less than 30% at 100 to 140 minutes, greater than 45% at 5 to 7 hours, and greater than 80% at 11 to 13 hours. In another type of embodiment, the formulation can be characterized by an in vitro dissolution profile providing release of the vitamin D compound of less than 30% at 2 hours, greater than 45% at 6 hours, and greater than 80% at 12 hours. In these types of embodiments, optionally, the release of the vitamin D compound is less than 60% at 5 to 7 hours, or less than 60% at 6 hours.

[0047] In another type of embodiment, the formulation can be characterized by an in vitro dissolution profile providing release of the vitamin D compound of about 20% to about 40% at 2 hours, at least 35% at 6 hours, and at least 70% at 12 hours. In another type of embodiment, the formulation can be characterized by an in vitro dissolution profile providing release of the vitamin D compound of about 25% to about 35% at 2 hours, at least 40% at 6 hours, and at least 75% at 12 hours. In these types of embodiments, optionally, the release of the vitamin D compound is, for example, 75% or less at 6 hours, or 65% or less at 6 hours, or 60% or less at 6 hours.

[0048] In any embodiment described herein, the stabilized formulation can be characterized by a t max , for example, in the range of 4 to 96 hours, or in the range of 18 to 30 hours, or in the range of 13 to 28 hours, or, for example, 28 hours.

[0049] In any embodiment contemplated herein, a formulation comprising 25-hydroxyvitamin D can be characterized as providing a baseline-adjusted C max .

[0050] In any method contemplated herein, such a method can include administering to a human patient a stabilized sustained release dosage form comprising a 25-hydroxyvitamin D compound, which includes administering to the patient an effective amount of the formulation to provide a baseline-adjusted C max , for example, in the range of about 0.2 to about 24 ng / mL.

[0051] In any of the methods contemplated herein, the method can comprise administering to a human patient a stabilized sustained release dosage form comprising a 25-hydroxyvitamin D compound, which comprises administering to the patient an effective amount of the formulation to provide a baseline-adjusted AUC of at least 52 ng h / mL and optionally less than 34,500 ng h / mL and further optionally about 12,000 ng h / mL or less 0-inf , for example in a range of about 52 ng h / mL to about 12,000 ng h / mL.

[0052] In any of the embodiments described herein, it is contemplated that the stabilized formulation can be bioequivalent to a freshly made product after storage. Thus, for example, the stabilized formulation can provide an area under the curve AUC (e.g., AUC 0-inf or AUC 0-t ) of active agent (or serum total 25-hydroxyvitamin D) within 90% confidence interval of the freshly made product, or within 80% to 125% of the mean, or within 80% to 120% of the mean, after storage. Additionally or in the alternative, the stabilized formulation can provide a maximum serum concentration C max (e.g., C max absolute value, or C max compared to baseline) of active agent (or serum total 25-hydroxyvitamin D) within 90% confidence interval of the freshly made product, or within 80% to 125% of the mean, or within 80% to 120% of the mean, after storage.

[0053] In one embodiment, the stabilized formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a wax matrix, and a cellulose compound. In one aspect, the stabilized formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a wax matrix, and a cellulose stabilizer. In another aspect, the formulation comprises one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3, a wax matrix, and an effective amount of a cellulose compound to provide a degree of stability as described herein, for example, in terms of the table just above or consistent with any of the examples described below. For example, such an amount can be effective to provide a difference of 30% or less between the amount of active agent released at a certain dissolution time point during in vitro dissolution after exposure to storage conditions of 25°C and 60% relative humidity for at least one month and the amount released at the same dissolution time point during in vitro dissolution conducted prior to exposing the formulation to the storage conditions, whereas a comparative formulation lacking the stabilizer would result in a greater difference in dissolution release after the same storage conditions.

[0054] In one aspect, the formulation is an improved formulation for controlled release of the vitamin D compound in the gastrointestinal tract of a subject who ingests the formulation. In one embodiment, the improvement comprises admixing a cellulose stabilizer into the formulation for controlled release of the vitamin D compound in the gastrointestinal tract of a subject who ingests the formulation. In another embodiment, the improvement comprises admixing into the formulation for controlled release of the vitamin D compound in the gastrointestinal tract of a subject who ingests the formulation an effective amount of a cellulose compound to provide a degree of stability as described herein, e.g., in terms of the table immediately above or consistent with any of the embodiments described below. For example, such an amount can be effective to provide a difference of 30% or less between the amount of active agent released at a certain point in time during in vitro dissolution after exposure to storage conditions of 25°C and 60% relative humidity for at least one month and the amount released at the same point in time during in vitro dissolution conducted prior to exposing the formulation to the storage conditions, whereas a comparative formulation lacking the stabilizer would result in a greater difference in dissolution release after the same storage conditions.

[0055] Stabilizing agents can include cellulose compounds. Examples of cellulose compounds and stabilizing agents for use in the stabilized formulations of the present disclosure can include, but are not limited to, cellulose uronic acid, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, polyanionic cellulose, and combinations thereof. Also contemplated are one or more of poloxamer (e.g., poloxamer 407), poly(ethylene oxide) polymers (e.g., Dow's POLYOX polymers), povidone, and fumed silica (e.g., AEROSIL 200, Evonik Industries AG, Essen, Germany). Stabilizing agents, such as cellulose compounds, are preferably present in an amount of at least about 5% (wt%) of the formulation, based on the total weight of the formulation, exclusive of any additional coatings or outer shells. For example, the cellulose compound can be present in an amount of at least 5 wt% of the formulation, or at least 10 wt% of the formulation, or at least 15 wt% of the formulation, or greater than 5 wt% of the formulation, or greater than 10 wt% of the formulation, or greater than 15 wt% of the formulation. Suitable ranges include 5 wt% to 30 wt%, 10 wt% to 20 wt%, 10 wt% to 15 wt%, 5 wt% to 15 wt%, and 7.5 wt% to 12.5 wt.%. Examples include about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, and about 15 wt%. It will be appreciated that the stabilizing agents referred to herein are agents that stabilize the dissolution release profile (and thus also the in vivo release profile) against substantial changes over time during storage conditions, such as typical shelf storage conditions. Other agents known in the art as preservatives to prevent degradation of the active components themselves are not intended to be encompassed within the term "stabilizing agent," although such preservatives are also contemplated for use in the formulations of the present application.

[0056] In one class of embodiments, the cellulose compound is a cellulose ether. Examples of cellulose ethers include, but are not limited to, methyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl methyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, and combinations thereof.

[0057] Hydroxypropyl methylcellulose (HPMC) is specifically contemplated. HPMC can be characterized by one or more of the following features, individually and in combination, that are specifically contemplated. The % methoxyl component in HPMC can be in the range of 19 to 24. The % hydroxypropyl component can be in the range of 7 to 12. The apparent viscosity (2% solution in water at 20 °C) can be at least 50,000 cP or at least 80,000 cP, or in the range of about 80 to 120,000 cP, or 3000 to 120,000 cP, or 11,000 to 120,000 cP, or 80,000 to 120,000 cP. In particular, the apparent viscosity (2% solution in water at 20 °C) can be in the range of 80,000 to 120,000 cP. The pH (1% solution in water) can be in the range of 5.5 to 8.0. For example, a suitable hydroxypropyl methylcellulose having all the foregoing properties, including an apparent viscosity (2% solution in water at 20 °C) in the range of 80,000 to 120,000 cP, is METHOCEL K100M CR (Dow Wolff Cellulosics, Midland, Michigan).

[0058] In one type of embodiment, the cellulose compound will be insoluble in the matrix formulation at the melting point of the primary component of the matrix, for example at 65 °C or in the range of 60 °C to 75 °C.

[0059] In one type of embodiment, the cellulose compound will be hydrophilic.

[0060] Pharmaceutical formulations according to the present disclosure comprising one or more of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3and a cellulose compound have unexpectedly improved stability compared to formulations lacking the cellulose compound. In one embodiment, the stabilized formulations according to the present disclosure comprise a mixture of an active agent-loaded lipophilic matrix comprising one or both of 25-hydroxyvitamin D2and 25-hydroxyvitamin D3and a cellulose stabilizer, wherein such formulations release an amount of 25-hydroxyvitamin D during in vitro dissolution that varies by 30% or less at any given dissolution time point compared to the amount released at the same dissolution time point during in vitro dissolution of a freshly manufactured product after exposure to storage conditions of 25 °C and 60% relative humidity for at least one month.

[0061] As shown in the examples below, formulations that are not stabilized exhibit a change in the amount of active ingredient released upon exposure of the composition to storage for a period of time. The amount of 25-hydroxyvitamin D released by a non-stabilized formulation after exposure to storage conditions can vary by more than 30% at a given dissolution time point compared to the amount released at the same dissolution time point during in vitro dissolution of a freshly made product. The variation can be an increase or decrease in the dissolution rate at a given time point, and such variations result in dissolution profiles that differ in shape from the initial dissolution profile. After storage as described herein, for example, after storage at 25°C and 60% RH for 3 months or more, non-stabilized formulations also exhibit different in vivo effects compared to stabilized formulations according to the present disclosure. After storage as described herein, for example, after storage at 25°C and 60% RH for 3 months or more, stabilized formulations exhibit different clinical pharmacokinetic parameters, such as improved bioavailability. Stabilized formulations according to the present disclosure can have a storage unstable base formulation combined with a stabilizer that facilitates storage stability of the formulation as described herein.

[0062] Matrices that releasably bind and controllably release active components can be, for example, lipophilic matrices, including wax matrices. Wax matrices can provide formulations that are solid or semi-solid at room temperature and solid, semi-solid, or liquid at body temperature, preferably semi-solid or liquid at body temperature. In one aspect, the wax matrix comprises a controlled release agent, an emulsifying agent, and an absorption enhancer.

[0063] Examples of controlled release agents suitable for use include, but are not limited to, waxes, including synthetic waxes, microcrystalline waxes, paraffin waxes, carnauba wax, and beeswax; polyethoxylated castor oil derivatives, hydrogenated vegetable oils, glyceryl monobehenate, glyceryl dibehenate, or glyceryl tribehenate; long chain alcohols, such as stearyl alcohol, cetyl alcohol, and polyethylene glycol; and mixtures of any of the foregoing. Non-digestible waxy materials, such as hard paraffin wax, are preferred.

[0064] The controlled release agent can be present in an amount of at least 5 wt% of the formulation or greater than about 5 wt% of the formulation. For example, depending on the controlled release agent used, the controlled release agent can be at least 5 wt% of the formulation, or at least 10 wt% of the formulation, or at least 15 wt% of the formulation, or at least 20 wt% of the formulation, or at least 25 wt% of the formulation, or greater than 5 wt% of the formulation, or greater than 10 wt% of the formulation, or greater than 15 wt% of the formulation, or greater than 20 wt% of the formulation, and / or greater than 25 wt% of the formulation. The controlled release agent can be present in an amount of 50 wt% or less, 40 wt% or less, 35 wt% or less, or 30 wt% or less. Suitable ranges include 5 wt% to 40 wt%, 10 wt% to 30 wt%, and 15 wt% to 25 wt%. Examples include about 15 wt%, about 16 wt%, about 17 wt%, about 18 wt%, about 19 wt%, about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, and about 25 wt%.

[0065] Examples of suitable emulsifiers for use in the formulation include, but are not limited to, lipophilic agents having an HLB of less than 7, such as mixed fatty acid monoglycerides; mixed fatty acid diglycerides; mixtures of fatty acid monoglycerides and diglycerides; lipophilic polyglycerol esters; glycerol esters, including glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; fatty acid glycerol lactate esters; propylene glycol esters, including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters, including sorbitan monostearate, sorbitan sesquioleate; fatty acids and their soaps, including stearic acid, palmitic acid, and oleic acid; and mixtures thereof; glyceryl monooleate, glyceryl dioleate, glyceryl monostearate, glyceryl distearate, glyceryl monopalmitate, and glyceryl dipalmitate; fatty acid glycerol lactate esters; propylene glycol esters, including propylene glycol monopalmitate, propylene glycol monostearate, and propylene glycol monooleate; sorbitan esters, including sorbitan monostearate, sorbitan sesquioleate; fatty acids and their soaps, including stearic acid, palmitic acid, and oleic acid; and mixtures thereof.

[0066] Preferred lipid agents are selected from glycerol esters and derivatives thereof. Preferred glycerol esters are selected from the group consisting of heavy or long chain glycerol esters, caprylocaproyl polyoxylglycerols, and mixtures thereof.

[0067] Preferred medium chain glycerol esters include, but are not limited to, medium chain monoglycerides, medium chain diglycerides, caprylic / capric triglycerides, glyceryl monolaurate, glyceryl monostearate, caprylocaproyl glycerols, glyceryl monocaprylate, glyceryl monocaprylate, glyceryl monocaprylate, caprylic / capric / linoleic triglycerides, and caprylic / capric / succinylated glycerides.

[0068] Glycerol monesters with low melting points are preferred for use in making the formulation. Preferred glycerol monesters include, but are not limited to, glyceryl monostearate, glyceryl monopalmitate, glyceryl monooleate, glyceryl monocaprylate, glyceryl monocaprate, glyceryl monolaurate, and the like, preferably glyceryl monostearate (GMS). GMS is a natural emulsifier. GMS is oil-soluble, but poorly soluble in water. GMS has an HLB value of 3.8. For example, the lipophilic emulsifier can be present in an amount ranging from about 10 wt% to about 40 wt% or about 20 wt% to about 25 wt%. Other examples include about 20 wt%, about 21 wt%, about 22 wt%, about 23 wt%, about 24 wt%, and about 25 wt%.

[0069] Examples of suitable absorption enhancers include, but are not limited to, caprylocaproyl macrogolglycerides, such as polyoxylated glycerides, also known as polyglycolized glycerides or PEGylated glycerides. PEGylated glycerides that can be employed in the composition include, but are not limited to, mixtures of glycerol monoesters, diesters, and triesters with mono- and di-esters of polyethylene glycol, polyoxylated almond glycerides, polyoxylated corn glycerides, and polyoxylated caprylic / capric triglycerides. The absorption enhancer can have an HLB value of 13 to 18 or 13 to 15.

[0070] One preferred absorption enhancer is known under the trade name GELUCIRE (Gattefosse Corporation, Paramus, New Jersey, USA). GELUCIRE is a well-known excipient that is a family of glycerol fatty acid esters and PEG esters, also known as polyglycolized glycerides. GELUCIRE is used in a variety of applications, including in the preparation of sustained release pharmaceutical compositions. GELUCIRE compounds are inert semi-solid waxy substances that are amphiphilic and are available with different physical characteristics, such as melting point, HLB, and solubility in various solvents. GELUCIRE is surface active in nature and disperses or dissolves in aqueous media to form micelles, microspheres, or vesicles. GELUCIRE is identified by its melting point / HLB value. Melting point is expressed in degrees Celsius. One or a mixture of different grades of GELUCIRE excipients can be selected to achieve the desired characteristics of melting point and / or HLB value. A preferred GELUCIRE composition is GELUCIRE 44 / 14, i.e., a mixture of lauroyl macrogolglycerides and lauroyl macrogoloxyglycerides having a melting point of 44°C and an HLB of 14. For example, the absorption enhancer can be present in an amount ranging from about 5 wt% to about 20 wt% or about 8 wt% to about 15 wt%. Other examples include about 8 wt%, about 9 wt%, about 10 wt%, about 11 wt%, about 12 wt%, about 13 wt%, about 14 wt%, and about 15 wt%.

[0071] The low melting point of the wax matrix provides a means of incorporating a pharmaceutically active ingredient, e.g., a vitamin D compound, such as 25-D2, 25-D3, or both, at a temperature of about 0 °C to about 50 °C above the melting point of the wax matrix, and then filling the melt (solution and / or dispersion) into a suitable capsule. The capsule can be any kind compatible with the temperature of filling of the melt, including soft or hard gelatin capsules, and animal or vegetable gelatin capsules. The melt solidifies within the capsule upon cooling to room temperature.

[0072] In one aspect, the stabilized formulation can further comprise an oily vehicle for the 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3. Any pharmaceutically acceptable oil can be used. Examples include animal (e.g., fish), vegetable (e.g., soybean), and mineral oils. The oil preferably will readily dissolve the 25-hydroxyvitamin D compound used. Preferred oily vehicles include non-digestible oils, such as mineral oil, particularly liquid paraffin, and squalene. For example, the oily vehicle can be present at a concentration ranging from about 10 wt% to about 50 wt%, or about 15 wt% to about 45 wt%, or about 20 wt% to about 40 wt%, or about 30 wt% to about 40 wt% of the formulation. In one type of embodiment, a suitable liquid paraffin can be characterized by one or more of the following parameters: specific gravity, about 0.88 to 0.89; kinematic viscosity (40 °C), about 64 cSt to about 70 cSt; molecular weight, 424; paraffin hydrocarbons %, about 59; and pour point, -24 °C. The ratio between the wax matrix and the oily vehicle can be optimized to achieve the desired release rate of the vitamin D compound. Thus, if a heavier oil component is used, relatively less wax matrix can be used, and if a lighter oil component is used, relatively more wax matrix can be used.

[0073] The stabilized controlled release composition according to the present application is preferably designed to contain 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3at a concentration of, for example, 1 to 1000 μg per unit dose, and is prepared in such a way as to achieve controlled or substantially constant release of the 25-hydroxyvitamin D2 / 25-hydroxyvitamin D3over an extended period of time, optionally into the ileum of the gastrointestinal tract of a human or animal. Exemplary doses include 1 μg to 1000 μg, 1 μg to 600 μg, 1 μg to 400 μg, 1 μg to 200 μg, 1 μg to 100 μg, 5 μg to 90 μg, 30 μg to 80 μg, 20 μg to 60 μg, 30 μg to 60 μg, 35 μg to 50 μg, 5 μg to 50 μg, and 10 μg to 25 μg, e.g., 20 μg, 25 μg, 30 μg, 40 μg, 50 μg, 60 μg, 70 μg, 80 μg, 90 μg, and 100 μg, per unit dose.

[0074] In a preferred class of embodiments, the controlled release formulation releases at least 70%, more preferably at least 80% of the vitamin D compound within the first 24 hours after administration.

[0075] Advantageously, 25-hydroxyvitamin D2, 25-hydroxyvitamin D3, or combinations thereof, can be administered with other therapeutic agents, e.g., orally, in dosages of, e.g., 1 to 100 μg per day, in accordance with the embodiments described above. In one type of embodiment, the dosage will be selected to provide an average increase in serum 25-hydroxyvitamin D3 of about 1 to 3 ng / ml over the dosing interval.

[0076] In embodiments, the formulations described herein can be administered to raise and preferably also maintain blood 1,25-dihydroxyvitamin D levels at 25 pg / mL, 30 pg / mL or higher, e.g., 25-65 pg / mL, for an extended period of time, e.g., at least one month, at least three months, at least six months, or longer.

[0077] In one aspect, the formulations described herein can be administered to a patient to lower or maintain a lowered serum parathyroid hormone level, preferably in an amount that reduces PTH levels by at least 30%, or in an amount required to reduce serum levels of PTH to the target range for CKD stage (e.g., 35-70 pg / mL (equivalent to 3.85-7.7 pmol / L) for stage 3, 70-110 pg / mL (equivalent to 7.7-12.1 pmol / L) for stage 4, and 150-300 pg / mL (equivalent to 16.5-33.0 pmol / L) for stage 5) defined in K / DOQI Guidelines No. 1.

[0078] In another aspect, the formulations according to the disclosure herein can be administered to a patient suffering from hyperparathyroidism secondary to chronic kidney disease (e.g., stage 3 or 4, or stage 3, 4, or 5) to lower serum PTH levels.

[0079] The dosages described herein are contemplated for use in any of the treatment methods described herein. It will be appreciated that the actual preferred amount of vitamin D compound in a particular case will vary according to the particular composition formulated, the mode of application, and the particular site being treated. Dosage can be determined using routine consideration, e.g., by routine comparison of the activity of the hormone with that of known agents, e.g., by means of appropriate routine pharmacological protocols.

[0080] The specific dose for each particular patient can depend on a variety of factors, e.g., age, body weight, general health, sex, diet, time and mode of administration, rate of excretion, and drugs used in combination and the severity of the particular condition for which the therapy is applied.

[0081] Patients in need of vitamin D supplementation include healthy subjects and subjects at risk of vitamin D insufficiency or deficiency, e.g., subjects with Stage 1, Stage 2, Stage 3, Stage 4, or Stage 5 CKD; infants, children, and adults who do not drink vitamin D fortified milk (e.g., subjects who are lactose intolerant, subjects who are allergic to cow's milk, vegans who do not drink milk, and breastfed infants); subjects with rickets; subjects with dark skin (e.g., in the United States, 42% of African American women between the ages of 15 and 49 are vitamin D deficient compared to 4% of white women); the elderly (whose ability to synthesize vitamin D is reduced and who are also more likely to be indoors); institutionalized adults (who can be indoors, including subjects with Alzheimer's disease or mental illness); subjects who cover all exposed skin (such as members of certain religions or cultures); subjects who always use sunscreen (e.g., application of sunscreen with a sun protection factor (SPF) of 8 reduces vitamin D production by 95%, and higher SPF values can further reduce vitamin D); subjects with fat malabsorption syndromes (including, but not limited to, cystic fibrosis, cholestatic liver disease, other liver disease, gallbladder disease, pancreatic enzyme deficiency, Crohn's disease, inflammatory bowel disease, celiac disease, or liposteatoteraphy, or surgery to remove part or all of the stomach and / or intestine); subjects with inflammatory bowel disease; subjects with Crohn's disease; subjects who have had a portion of the small intestine removed; subjects with gum disease; subjects taking medications that increase the catabolism of vitamin D, including phenytoin, fosphenytoin, phenobarbital, carbamazepine, and rifampin; subjects taking medications that decrease the absorption of vitamin D, including cholestyramine, colestipol, orlistat, mineral oil, and fat substitutes; subjects taking medications that inhibit the activation of vitamin D, including ketoconazole; subjects taking medications that decrease calcium absorption, including corticosteroids; obese subjects (there is less bioavailability of vitamin D deposited in body fat stores); subjects with osteoporosis; patients with low bone mineral density and osteoporosis; and / or postmenopausal women.According to the Institute of Medicine's report on Dietary Reference Intakes for vitamin D, food consumption data indicate that median intakes of vitamin D are below the current recommendations for both younger and older women; data indicate that more than 50% of both younger and older women do not consume the recommended amount of vitamin D.

[0082] Treatment of subjects suffering from renal osteodystrophy, including osteomalacia and osteitis fibrosa cystica, is optionally excluded from the methods of the application described herein.

[0083] In other aspects, the compositions and methods of the application are useful in the prophylactic or therapeutic treatment of a vitamin D-responsive disease, i.e., a disease in which vitamin D, 25-hydroxyvitamin D, or active vitamin D (e.g., 1,25-dihydroxyvitamin D) prevents the onset or progression of the disease, or reduces the signs or symptoms of the disease. Such vitamin D-responsive diseases include cancer (e.g., breast cancer, lung cancer, skin cancer, melanoma, colon cancer, colorectal cancer, rectal cancer, prostate cancer, and bone cancer). 1,25-dihydroxyvitamin D has been observed to induce cell differentiation and / or inhibit cell proliferation in a number of cells in vitro. Vitamin D-responsive diseases also include autoimmune diseases, e.g., Type I diabetes mellitus, multiple sclerosis, rheumatoid arthritis, polymyositis, dermatomyositis, scleroderma, fibrosis, Grave's disease, Hashimoto's disease, acute or chronic transplant rejection, acute or chronic graft-versus-host disease, inflammatory bowel disease, Crohn's disease, systemic lupus erythematosus, Sjogren's Syndrome, eczema, and psoriasis; dermatitis, including atopic dermatitis, contact dermatitis, allergic dermatitis, and / or chronic dermatitis. Vitamin D-responsive diseases also include other inflammatory diseases, e.g., asthma, chronic obstructive pulmonary disease, polycystic kidney disease, polycystic ovary syndrome, pancreatitis, nephritis, hepatitis, and / or infection. Vitamin D-responsive diseases have also been reported to include hypertension and cardiovascular disease. Thus, the present application contemplates prophylactic or therapeutic treatment of a subject at risk of or suffering from cardiovascular disease, e.g., a subject having atherosclerosis, arteriosclerosis, coronary artery disease, cerebrovascular disease, peripheral vascular disease, myocardial infarction, myocardial ischemia, cerebral ischemia, stroke, congestive heart failure, cardiomyopathy; obesity or other body weight abnormalities; lipid abnormalities (e.g., hyperlipidemia; lipid metabolism abnormalities, including associated diabetic lipid metabolism abnormalities and mixed lipid metabolism abnormalities; hypoalphalipoproteinemia; hypertriglyceridemia; hypercholesterolemia; and low HDL (high density lipoprotein)); metabolic abnormalities (e.g., metabolic syndrome; Type II diabetes mellitus; Type I diabetes mellitus; hyperinsulinemia; impaired glucose tolerance; insulin resistance; diabetic complications, including neuropathy, nephropathy, retinopathy, diabetic foot ulcer, and cataracts); and / or thrombosis.

[0084] Disease conditions that can benefit from the level modulation of vitamin D compounds include, but are not limited to: (i) in the parathyroid gland - hypoparathyroidism, pseudohypoparathyroidism, secondary hyperparathyroidism; (ii) in the pancreas - diabetes; (iii) in the thyroid - medullary carcinoma; (iv) in the skin - psoriasis, wound healing; (v) in the lung - sarcoidosis and tuberculosis; (vi) in the kidney - chronic renal disease, hypophosphatemic VDRR, vitamin D dependent rickets; (vii) in the bone - anticonvulsant therapy, osteofibrosis dysplasia, osteitis fibrosa cystica, osteomalacia, osteoporosis, osteopenia, osteosclerosis, renal osteodystrophy, rickets; (viii) in the intestine - glucocorticoid antagonism, idiopathic hypercalcemia, malabsorption syndrome, steatorrhea, tropical sprue; and (ix) autoimmune disorders.

[0085] In embodiments of the application, the disease condition that benefits from the level modulation of vitamin D compounds is selected from cancer, a skin disorder (e.g., psoriasis), a parathyroid disorder (e.g., hyperparathyroidism and secondary hyperparathyroidism), a bone disorder (e.g., osteoporosis), and an autoimmune disorder.

[0086] The formulations can be prepared by procedures well within the ability of the ordinarily skilled artisan. For example, the matrix components (e.g., waxes and oily vehicles) can be melted, if necessary, to provide a flowable liquid, making it easier to obtain a homogeneous mixture. The active agent (e.g., 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3) is added to the liquid vehicle, e.g., dissolved in an alcohol such as absolute ethanol, and the ingredients are mixed to provide a homogeneous mixture. In one type of embodiment, the stabilizing agent can be added after all of the matrix components (e.g., waxes and oils) are blended and before combination with the active agent. The mixture can be cooled and stored before being divided into unit dosage forms, such as filled gelatin capsules, at a later time.

[0087] In one type of method, a portion of the oily vehicle, the controlled release agent, and the emulsifying agent are heated to a relatively high temperature (e.g., 65 °C) and mixed prior to the addition of the absorption enhancer, followed by additional mixing until homogeneous, and then cooled to an intermediate high temperature (e.g., 50 °C to 55 °C). In a separate container, the antioxidant preservative and the remaining oily vehicle are mixed and heated to an intermediate high temperature (e.g., 50 °C), then combined with the wax mixture and mixed until a homogeneous solution is obtained. Next, the stabilizing agent is added while mixing. Next, the solution of the vitamin D compound in alcohol is combined with the homogeneous waxy solution, mixed until a homogeneous solution is obtained, preferably filled into a capsule, and then cooled to room temperature. In another preferred method, a portion of the oily vehicle, the controlled release agent, and the emulsifying agent are heated to a temperature of 55 °C to 60 °C and mixed prior to the addition of the absorption enhancer, followed by additional mixing until homogeneous. In a separate container, the antioxidant preservative, the remaining oily vehicle, and the stabilizing agent are mixed and heated to a temperature of 55 °C to 60 °C, then combined with the wax mixture and mixed until a homogeneous solution is obtained. Next, the solution of the vitamin D compound in alcohol is combined with the homogeneous waxy solution, mixed until a homogeneous solution is obtained, preferably filled into a capsule, and then cooled to room temperature.

[0088] The formulation is preferably placed in a capsule prior to administration to a patient in need of treatment. Such capsules can be hard or soft, and soft capsules are particularly contemplated. The formulation can be filled into gelatin capsules using a standard capsule filling machine, such as by melting the formulation and injection filling it into a soft capsule shell. Exemplary soft capsule shells include VEGICAPS and OPTISHELL technology (Catalent, Somerset, NJ, USA). In the alternative, the formulation can be made into a unit dosage form by any other suitable process, for example to give tablets, sachets, dragees, suppositories, and the like.

[0089] In one type of embodiment, the formulation is prepared and administered by oral delivery. In another type of embodiment, the formulation is prepared and administered as a suppository, for example a rectal suppository.

[0090] Unless otherwise specified, the formulations, as well as the methods of use and manufacture, are contemplated to include embodiments comprising any combination of one or more of the additional optional elements, features, and steps described further below.

[0091] Thus, in one type of embodiment, the formulation further comprises a preservative, such as an antioxidant. Butylated hydroxytoluene (BHT) is preferred.

[0092] In another type of embodiment, the vitamin D compound is administered in combination with one or more other therapeutic agents.

[0093] If the vitamin D compound is administered in combination with one or more other therapeutic agents, the identity of each compound in the combination administered will depend on the particular disease condition being treated. For example, we can choose to administer 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3(e.g., orally) with one or more calcium salts (intended as calcium supplements or dietary phosphate binders), bisphosphonates, calcimimetics, niacin, iron, phosphate binders, cholecalciferol, ergocalciferol, active vitamin D sterols, glycemic and hypertension control agents, various antineoplastic agents, and inhibitors of CYP24 and other cytochrome P450 enzymes that can degrade vitamin D agents. In addition, we can choose to administer 25-hydroxyvitamin D2and / or 25-hydroxyvitamin D3intravenously with cholecalciferol, ergocalciferol, active vitamin D sterols, glycemic and hypertension control agents, various antineoplastic agents, and inhibitors of CYP24 and other cytochrome P450 enzymes that can degrade vitamin D agents. In fact, higher doses of the compounds of the present application are used where the therapeutic treatment of a disease condition is the desired goal, while lower doses are generally used for prophylactic purposes, it will be appreciated that the particular dose administered in any given case will be adjusted in accordance with the particular compound administered, the disease being treated, the condition of the subject, and other relevant medical circumstances that can alter the activity of the drug or the response of the subject, as is well known to those skilled in the art.

[0094] As described above, the formulation is preferably filled into gelatin capsules, but it can also be administered in pure form, or with one or more outer coating layers, such as an enteric coating. It is also contemplated that the formulation can be compressed into tablets, and in such cases can include one or more tabletting excipients.

[0095] In the compositions and methods described herein, the preferred steps, preferred components, preferred ranges of their compositions, and preferred combinations of the foregoing can be selected from the various specific examples provided herein. For example, a preferred formulation includes 25-hydroxyvitamin D (e.g., about 30 μg, about 60 μg, or about 90 μg 25-hydroxyvitamin D3), about 2 wt% (e.g., 2.32 wt%) absolute ethanol, about 10 wt% (e.g., 9.75 wt%) GELUCIRE 44 / 14, about 20 wt% (e.g., 20.00 wt.%) hard paraffin, about 23 wt% (e.g., 22.55 wt%) GMS, about 35 wt% (e.g., 35.36 wt%) liquid paraffin or mineral oil, about 10 wt% HPMC, and optionally, a small amount of preservative (e.g., 0.02 wt% BHT). Variations of this formulation will include about 15% (e.g., 15.29 wt%) HPMC and about 30 wt% (e.g., 29.88 wt%) liquid paraffin or mineral oil.

[0096] Example

[0097] The following examples illustrate particular formulations and methods of making them. These examples are provided for illustration and are not intended to limit the scope of the application.

[0098] In vitro dissolution testing in the examples was performed using USP Apparatus 2 (paddle method) as described in USP 29-NF 24 General Chapter <711> Dissolution, using the dissolution media described below. In general, the method was performed according to the following steps. A specified volume of dissolution medium (± 1%) was placed in the vessel of the designated apparatus, the apparatus was assembled, the dissolution medium was equilibrated to 37 ± 0.5 °C, and the thermometer was removed. The dosage unit was placed in the apparatus, air bubbles were carefully excluded from the surface of the dosage unit, and the apparatus was immediately operated at the designated rate. At each specified time, a sample was withdrawn from the mid- section area between the surface of the dissolution medium and the top of the rotating paddle, not less than 1 cm from the wall of the vessel. The aliquot withdrawn for analysis was replaced with an equal volume of fresh dissolution medium at 37 °C. The duration of the test was maintained with the vessel covered, and the temperature of the mixture under test was checked at appropriate times. Analysis was performed using an appropriate detection method, in this case ultra-performance liquid chromatography (UPLC).

[0099] Six capsules of each formulation were tested at each time point. The dissolution medium was 0.05 M pH 6.8 phosphate buffer / 1% sodium lauryl sulfate dissolution medium (37 ± 0.5 °C), and the apparatus was operated at 100 revolutions per minute. Samples were taken at 2, 4, 6, 8, and 12 hours, and the 25-hydroxyvitamin D content of each sample was determined using UPLC.

[0100] Example 1 - In vitro dissolution of unstabilized sustained release formulations of 25-hydroxyvitamin D

[0101] A formulation made from a mixture of 90 μg 25-hydroxyvitamin D3, 19.98 wt% hard paraffin, 37.85 wt% GMS, 9.76 wt% GELUCIRE 44 / 14, 2.36 wt% absolute ethanol, 29.88 wt% liquid paraffin, and 0.02 wt% BHT (Comparative Formulation 1) was tested for dissolution. This formulation did not include a cellulose compound. The average amount of 25-hydroxyvitamin D3 released at T=0 and after the formulation was controlled storage at 5 °C and ambient humidity for up to 12 months was calculated as the average percentage of the nominal drug load per dosage form (average labeled content %, LC%) and is summarized in the table below. It was determined that samples should be stored for a period of about 3 months at a temperature in the range of 15 °C to 30 °C and ambient humidity prior to testing. Therefore, samples that should be representative of time zero are labeled T=0 p(pseudo-time zero) and it will be appreciated that the nominally 1 month, 3 month, 6 month, 9 month, and 12 month aged samples also experience the approximately 3 month aging period just described. To provide a more accurate baseline, a fresh batch of the same type of sample was prepared and tested without any aging; this data is labeled T=0 f Indicating fresh sample. The coefficient of variation (CV%) is also reported. From the T=0 p and T=0 f The percent change in the initial amount of 25-hydroxyvitamin D3 released from the batch is provided in square brackets and double square brackets, respectively.

[0102]

[0103]

[0104] Dissolution of Comparative Formulation 1 was tested after storage at 25°C and 60% relative humidity for 0 to 12 months. The results are summarized in the table below.

[0105]

[0106]

[0107]

[0108] Dissolution of Comparative Formulation 1 was tested after storage at 40°C and 75% relative humidity for 0, 1, 3, and 6 months. The results are summarized in the table below.

[0109]

[0110]

[0111] Without intending to be bound by any particular theory, it is believed that the increase in the degree of dissolution compared to the pseudo T=0 values after storage at 40°C is due to a combination of the aging effects described above for the pseudo T=0 samples tested and the temperature-dependent phase transition of the formulation upon storage at 40°C.

[0112] Aged product according to Comparative Formulation 1 was heat-cured and then tested for dissolution. Curing consists of the application of heat treatment and has been shown to stabilize pharmaceutical formulations (see, e.g., U.S. Patent No. 6,645,527). Comparative Formulation 1 (aged sample) was heated at 40°C for 72 hours to cure, and then stored at room temperature for 8 weeks. Release of 25-hydroxyvitamin D3 from the cured formulation was tested after storage at room temperature for 0, 2, 4, and 8 weeks. The results are summarized in the table below.

[0113]

[0114]

[0115] Example 2 - In vitro dissolution of stabilized controlled release formulations of 25-hydroxyvitamin D

[0116] The dissolution of a sustained release formulation comprising 90 μg 25-hydroxyvitamin D3, 19.88 wt% hard paraffin, 15.29 wt% hydroxypropyl methylcellulose, 22.55 wt% GMS, 9.76 wt% GELUCIRE 44 / 14, 2.36 wt% absolute ethanol, 29.88 wt% liquid paraffin, and 0.02 wt% BHT (Example Formulation A) was tested after storage at room temperature for 0 to 11 weeks. The results are summarized in the table below.

[0117]

[0118]

[0119] The dissolution of a sustained release formulation comprising 90 μg 25-hydroxyvitamin D3, 19.88 wt% hard paraffin, 10.00 wt% hydroxypropyl methylcellulose, 22.55 wt% GMS, 9.76 wt% GELUCIRE 44 / 14, 2.36 wt% absolute ethanol, 35.17 wt% liquid paraffin, and 0.02 wt% BHT (Example Formulation B) was tested after storage at room temperature for 0 to 26 weeks. The results are summarized in the table below.

[0120]

[0121] Example Formulation B exhibited a substantially stable dissolution profile after storage at room temperature for at least 26 weeks.

[0122] The stability of stabilized formulations comprising 30 μg (Example Formulation C), 60 μg (Example Formulation D), or 90 μg (Example Formulation E) of 25-hydroxyvitamin D3 was tested using storage conditions of 25 °C and 60% relative humidity and 40 °C and 75% relative humidity. The compositions of Example Formulations C to E are summarized in the table below:

[0123]

[0124]

[0125] The formulations exhibited a substantially stable dissolution profile after storage at 25 °C and 60% relative humidity for at least 24 months (Figure 1). The dissolution results (LC% and CV%) are summarized in the table below.

[0126]

[0127]

[0128] 4 repeats instead of 6

[0129] The percent change between the amount of 25-hydroxyvitamin D3 released after aging compared to the initial amount released is summarized in the table below.

[0130]

[0131] Example Formulations C-E also exhibited substantially stable dissolution profiles after storage at 40 °C and 75% RH for at least 6 months (Figure 2). Dissolution results are summarized in the table below.

[0132]

[0133]

[0134]

[0135] The percent change between the amount of 25-hydroxyvitamin D3 released after aging compared to the initial amount released is summarized in the table below.

[0136]

[0137]

[0138] Stability of Comparative Formulation 1, which does not contain a cellulose compound, and Example Formulation E, which contains hydroxypropyl methylcellulose, was evaluated after storage at 25 °C and 60% relative humidity for 12 months Figure 3 ). Dissolution results are summarized in the table below.

[0139]

[0140] The percent change between the amount of 25-hydroxyvitamin D3 released after aging compared to the initial amount released is summarized in the table below.

[0141]

[0142] Example 3: In vivo results for unstabilized and stabilized controlled release formulations

[0143] In vivo studies were conducted to evaluate the clinical pharmacokinetics of unstable and stable controlled-release formulations of 25-hydroxyvitamin D3 in human subjects. In Study A, 28 subjects with stage 3 or 4 CKD, secondary hyperparathyroidism (stage 3: 70–1000 pg / mL iPTH; stage 4: 110–1000 pg / mL iPTH), and vitamin D deficiency (baseline serum total 25-hydroxyvitamin D of 15 ng / mL to 29 ng / mL) received a single oral dose of controlled-release capsules containing 450 μg or 900 μg of 25-hydroxyvitamin D3, 20.00 wt% paraffin, 37.85 wt% GMS, 9.75 wt% GELUCIRE 44 / 14, 2.32 wt% anhydrous ethanol, 30.06 wt% mineral oil, and 0.02 wt% BHT (comparative formulation 3) or a single intravenous dose of 448 μg of 25-hydroxyvitamin D3 in an ethanol solution. Neither formulation contained a cellulose compound.

[0144] Serum concentrations of 25-hydroxyvitamin D3 gradually increased after administration of the oral dose. The increase in 25-hydroxyvitamin D3 was dose-proportional and reached an approximate mean maximum observed serum concentration of 32 ng / mL after administration of 900 μg capsules (C0.05). max C max The time to onset (Tmax) was approximately 13 hours after administration. In contrast, the concentration of 25-hydroxyvitamin D3 increased rapidly after intravenous administration. Peak serum levels were reached immediately after intravenous administration (Tmax = 0.5 hours) and an approximate mean concentration of 134 ng / mL was achieved. max The bioavailability of oral doses is approximately 6% to 11%. The terminal half-life (t) of 25-hydroxyvitamin D3 after oral administration is... 1 / 2 The treatment duration was approximately 12 to 22 days. No adverse effects on serum calcium or phosphorus, or urinary calcium were observed in any treatment group.

[0145] Following intravenous administration, mean serum total 1,25-dihydroxyvitamin D increased rapidly, increasing by approximately 13 pg / mL from pretreatment baseline 6 hours after administration. In contrast, following administration of 900 μg capsules, mean serum total 1,25-dihydroxyvitamin D increased gradually and proportionally to the dose, increasing by approximately 7 pg / mL 48 hours after administration.

[0146] No significant changes in serum iPTH were observed in the initial 96 hours following intravenous administration. In contrast, for subjects receiving 900 μg capsules, serum PTH gradually decreased after administration, reaching a maximum inhibition of approximately 20% from pre-treatment baseline. Observed pharmacokinetic parameters for all treatment groups are summarized in the table below.

[0147]

[0148]

[0149] Baseline-adjusted pharmacokinetic parameters for all treatment groups are summarized in the table below.

[0150]

[0151]

[0152] In Study B, 20 healthy subjects with a mean baseline serum 25-hydroxyvitamin D of about 24 ng / mL (range 11 ng / mL to 45 ng / mL) received a single oral dose of stabilized controlled release capsules containing 900 μg 25-hydroxyvitamin D3, 20.00 wt% hard paraffin, 10.00 wt% HPMC, 22.55 wt% GMS, 9.75 wt% GELUCIRE 44 / 14, 2.32 wt% absolute ethanol, 35.36 wt% mineral oil, and 0.02 wt% BHT (Example Formulation F) or a single intravenous dose of 448 μg 25-hydroxyvitamin D3 in ethanol solution.

[0153] The gradual increase in 25-hydroxyvitamin D3 levels was demonstrated by the prolonged Tmaxcompared to the intravenous dose after administration of the stabilized oral formulation. The pharmacokinetic profile after administration of the stabilized oral formulation demonstrated a gradual increase in 25-hydroxyvitamin D3 concentrations with a mean Tmaxof 28 hours, avoiding rapid increases in blood levels in most subjects. Administration of the intravenous dose resulted in a rapid increase in 25-hydroxyvitamin D3 concentrations in all subjects. Avoiding rapid increases in 25-hydroxyvitamin D3 levels was highlighted by the significant differences observed between treatment groups in the C max max max

[0154] Despite being about twice as high, exposure to 25-hydroxyvitamin D3 after administration of the controlled release capsules was about twice as low as after the intravenous dose, resulting in a bioavailability of about 25%.t 1 / 2 1 / 2 The values for t 1 / 2 and CL are consistent with the reported prolonged elimination of 25-hydroxyvitamin D3. In addition, the Vdvalues suggest that 25-hydroxyvitamin D3 is maintained in systemic circulation, possibly highly bound to DBP. The observed pharmacokinetic parameters for all treatment groups are summarized in the table below.​​

[0155]

[0156] Baseline-adjusted pharmacokinetic parameters for all treatment groups are summarized in the table below.

[0157]

[0158]

[0159] Mean of individual subject bioavailability

[0160] 缩 Write: WA, not applicable

[0161] The study demonstrated that the stabilized controlled release formulation altered the rate of absorption of 25-hydroxyvitamin D3, resulting in a more gradual increase in serum 25-hydroxyvitamin D3 levels while maintaining the distribution and elimination characteristics. The stabilized formulation demonstrated improved pharmacokinetic parameters, such as increased Tmax, AUC, and bioavailability, compared to the unstabilized formulation of the same dose in Study A.

[0162] In Study C, 78 subjects with stage 3 CKD (eGFR 25-70 mL / min / 1.73 m 2 ), SHPT (>70 pg / mL plasma iPTH), and vitamin D insufficiency (serum total baseline 25-hydroxyvitamin D of 10 ng / mL to 29 ng / mL) received a daily oral dose of a stabilized controlled release formulation comprising 30 pg, 60 pg, or 90 pg 25-hydroxyvitamin D3, 20.00 wt% hard paraffin, 10.00 wt% HPMC, 22.55 wt% GMS, 9.75 wt% GELUCIRE 44 / 14, 2.32 wt% absolute ethanol, 35.36 wt% mineral oil, and 0.02 wt% BHT (Example Formulations C, D, and E from Example 2) or placebo for 6 weeks.

[0163] Mean baseline serum 25-hydroxyvitamin D3 concentrations were comparable across treatment groups and ranged from about 16 to 20 ng / mL. Following treatment with 25-hydroxyvitamin D3, mean levels of serum 25-hydroxyvitamin D3 gradually and in a dose-proportional manner increased following repeated daily administration of 25-hydroxyvitamin D3 and approached steady state (Cmax) by Week 6. Figure 4 ) for the groups administered 30 pg, 60 pg, and 90 pg 25-hydroxyvitamin D3, respectively, were about 28, 60, and 86 ng / mL, respectively. As background-adjusted AUC max values were about 28, 60, and 86 ng / mL, respectively. As background-adjusted AUC 0-6周The mean exposure to 25-hydroxyvitamin D3 assessed across the dose groups was proportional to dose. Mean serum 25-hydroxyvitamin D3 levels declined slowly after the last dose, but remained above baseline for all groups through the end of the study. Mean t 1 / 2 was between about 25 and 50 days. Baseline-adjusted pharmacokinetic parameters for 25-hydroxyvitamin D3 are summarized in the table below.

[0164]

[0165] The mean baseline serum 1,25-dihydroxyvitamin D concentration across the treatment groups was comparable and increased gradually, similar to the effect on serum 25-hydroxyvitamin D3 concentration. Mean ± SD baseline-adjusted C max values were higher in the 60 μg and 90 μg groups (18.4 ± 6.24 and 19.9 ± 14.30 ng / mL, respectively) compared to placebo and 30 μg groups (5.7 ± 6.35 and 6.4 ± 7.66 ng / mL, respectively). As baseline-adjusted AUC 0-6周 The mean exposure to 1,25-dihydroxyvitamin D assessed across the 25-hydroxyvitamin D3 dose groups was proportional to dose. Baseline-adjusted pharmacokinetic parameters for 1,25-dihydroxyvitamin D are summarized in the table below.

[0166]

[0167] The stabilized controlled-release formulation of 25-hydroxyvitamin D3 increased serum total 25-hydroxyvitamin D levels to > 30 ng / mL in a significantly greater number of subjects in all active dose groups compared to placebo. Similarly, the stabilized formulation significantly reduced mean plasma iPTH from baseline compared to placebo.

[0168] Administration of 25-hydroxyvitamin D3in a stabilized controlled-release formulation increased mean serum total 25-hydroxyvitamin D proportionally to the dose administered each day. The lowest administered dose (30 μg) increased serum total 25-hydroxyvitamin D from pre-treatment baseline (21.7 ± 1.8 ng / mL) by 15.6 ± 1.7 (SE) at the end of treatment, and the highest dose (90 μg) increased serum total 25-hydroxyvitamin D from 21.8 ± 1.2 ng / mL by 61.1 ± 6.1 ng / mL. In contrast, a decrease of 1.2 ± 0.7 ng / mL was observed at the end of treatment in the combination placebo group. The difference between the treatment and placebo groups was significant (p < 0.0001) for all three dose levels studied. The mean serum 25-hydroxyvitamin D level in the 30 μg dose group at the end of treatment was 37.3 ± 1.8 ng / mL (slightly above the K / DOQI designated minimum adequate level of 30 ng / mL), indicating that 30 μg was the minimum effective dose.

[0169] The percentage of treated subjects who achieved serum total 25-hydroxyvitamin D levels > 30 ng / mL at the end of treatment was 92.3%, 100.0%, and 100.0% in the 30 μg, 60 μg, and 90 μg dose groups, compared to 0% in placebo. These differences in response rate between active agent and placebo treatment were all significant (p < 0.001).

[0170] Mean plasma iPTH decreased proportionally to the administered dose of 25-hydroxyvitamin D3at the end of treatment. The lowest administered dose (30 μg) decreased iPTH from pre-treatment baseline by 20.2 ± 5.8 (SE) %, and the highest dose (90 μg) decreased iPTH by 35.9 ± 4.2%. An increase of 17.2 ± 7.8% was observed at the end of treatment in the combination placebo group. The difference between the groups receiving 25-hydroxyvitamin D3and placebo was significant (p < 0.005) for all three dose levels studied, and is favorably compared to the differences observed with more potent and calcemic oral vitamin D hormone replacement therapies (e.g., doxercalciferol, paricalcitol, and calcitriol) used in placebo-controlled studies for longer treatments.

[0171] The percentage of subjects receiving 25-hydroxyvitamin D3 who achieved a confirmed reduction (i.e., two consecutive measurements) in iPTH of at least 20% or 30% from pre-treatment baseline at EOT increased with dose up to 60 μg. Similar response rates were observed in the 60 μg and 90 μg treatment groups, indicating that no further benefit in iPTH reduction was observed for the 90 μg dose in this study. The response rates for a confirmed 20% reduction in iPTH were 38.5%, 70.6%, and 76.5% for the 30 μg, 60 μg, and 90 μg dose groups, respectively, compared to 9.7% in the placebo group. The difference in response rates observed for a 20% reduction was only significant (p < 0.005) in the 60 μg and 90 μg dose groups and for a 30% reduction in all three dose groups (p < 0.05). This data supports the conclusion that 30 μg of 25-hydroxyvitamin D3 per day in a stabilized controlled release formulation is the minimum effective dose.

[0172] The stabilized formulation of 25-hydroxyvitamin D3 had no clinically significant effects on albumin-corrected serum calcium, serum phosphorus, and urinary calcium excretion. There were no adverse effects on serum calcium or serum phosphorus or urinary calcium during the 6-week treatment period.

[0173] Pharmacokinetic analysis revealed that the stabilized formulation of 25-hydroxyvitamin D3 increased 25-hydroxyvitamin D3 exposure (AUC and C max ), where t 1 / 2 There were no differences after 6 weeks of administration. However, steady state modeling showed that steady state would have been achieved in all dose groups by week 7-9.

[0174] The data from this study clearly demonstrate that the stabilized controlled release formulation of 25-hydroxyvitamin D3 effectively raises serum total 25-hydroxyvitamin D to a minimum adequate level of 30 ng / mL and reduces plasma iPTH. The study also shows that the stabilized formulation of 25-hydroxyvitamin D3 has no clinically meaningful effects on serum calcium or phosphorus at the doses investigated.

[0175] Example 4: Pharmacokinetic and pharmacodynamic profile of modified release calcifediol in CKD subjects with secondary hyperparathyroidism and vitamin D deficiency

[0176] A multicenter, randomized, double-blind, placebo-controlled, repeat-dose, safety, efficacy, and PK / PD study of stabilized, sustained-release 25-hydroxyvitamin D3 (calcifediol, 25D3) capsules was conducted in subjects in 2 cohorts. Subjects with stage 3 CKD (eGFR of 25-70 mL / min / 1.73 m 2Male and female subjects, aged 18 to 85 years, with vitamin D insufficiency (serum 25-hydroxyvitamin D > 10 and < 29 ng / mL), SHPT (plasma iPTH > 70 pg / mL), and not requiring regular hemodialysis for this study. Subjects who met the criteria in the first cohort were randomized in a 1 : 1 : 1 ratio into 3 treatment groups: 2 groups received 60 or 90 μg daily oral doses of capsules, respectively, and 1 group received matching placebo capsules. Subjects in Cohort 2 were randomized in a 1 : 1 ratio into 2 treatment groups: 1 group received 30 μg capsules daily and the other received placebo. Subjects in each cohort completed a 6-week treatment and entered a 6-week follow-up period during which PK and PD samples were collected weekly. Serum calcium (Ca), phosphorus (P), 25D3, total 1,25-dihydroxyvitamin D (1,25D), and plasma iPTH were monitored weekly during the 6-week treatment and 6-week follow-up. ANCOVA models examined the association of 25D3 exposure with changes from baseline in 1,25D and iPTH. Covariates included baseline eGFR, body weight and height, sex, age, race, diabetes status, and baseline concentration of 1,25D or iPTH.

[0177] Figure 4 The resulting mean baseline-adjusted calcifediol concentrations for the treatment groups (PK population) are shown. The mean levels of serum calcifediol increased gradually and in a dose-proportional manner and approached steady state by Week 6. After 6 weeks of follow-up, the levels decreased in all active treatment groups but remained higher than baseline.

[0178] Figure 5 The resulting summary baseline-adjusted PK parameters for calcifediol concentrations for the treatment groups (PK population) are shown.

[0179] Figure 6 The resulting mean baseline-adjusted serum 1,25-dihydroxyvitamin D levels during the 6-week treatment (PK population) are shown. Mean baseline-adjusted serum total 1,25-dihydroxyvitamin D levels increased over time in those subjects administered active capsules compared to those administered placebo.

[0180] Figure 7 The resulting summary baseline-adjusted repeated-dose PK parameters for serum 1,25-dihydroxyvitamin D for the treatment groups (PK population) are shown.

[0181] Figure 8 The resulting mean baseline percent change in plasma iPTH levels during the 6-week treatment (PK population) are shown. Active capsules significantly reduced mean plasma iPTH from baseline by 21%, 33%, and 39% in all dose groups (30, 60, and 90 μg, respectively) compared to the 17% increase in the combined placebo group.

[0182] Figure 9 Summary of resultant baseline-adjusted repeated-dose PK parameters for plasma iPTH are shown for the treatment groups (PK population).

[0183] Figure 10 and 11 Percent change from baseline in plasma iPTH at EOT versus baseline-adjusted calcifediol and 1,25-dihydroxyvitamin D exposures (AUC 0-6周 ) in the PK population. The percent decrease in plasma iPTH from baseline to EOT increased with increasing serum calcifediol and total 1,25-dihydroxyvitamin D exposures (expressed as baseline-adjusted AUC 0-6周 ) during treatment.

[0184] Stabilized, sustained-release 25-hydroxyvitamin D3 capsules normalized 25D levels in most subjects and significantly reduced iPTH in all dose groups (30, 60, and 90 μg). Stabilized, sustained-release 25-hydroxyvitamin D3 capsules gradually increased serum 25D3 and serum 1,25D levels in a dose-dependent increase in exposure. Both 25D3 and total 1,25D exposures were significantly and inversely correlated with the change in plasma iPTH from baseline. Only eGFR was a significant covariate in both models. These findings demonstrate that stabilized, sustained-release 25-hydroxyvitamin D3 capsules reliably normalized 25D levels, increased serum 1,25D levels, and suppressed elevated plasma iPTH without clinically meaningful effects on serum Ca and P at the doses investigated.

[0185] ***

[0186] The foregoing description is given for clearness of understanding only, and no unnecessary limitations should be understood therefrom, as modifications within the scope of the invention can be apparent to those having ordinary skill in the art.

[0187] Throughout this specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to mean the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0188] Throughout the specification, unless otherwise indicated, where a composition is described as including a component or substance, it is contemplated that the composition can, in some embodiments, consist essentially of, or consist of, any combination of the recited components or substances, unless otherwise specified. Likewise, where a method is described as including a particular step, it is contemplated that, in some embodiments, the method can, in some embodiments, consist essentially of, or consist of, any combination of the recited steps, unless otherwise specified. The inventive subject matter disclosed herein can be implemented in the absence of any element or step not specifically disclosed herein.

[0189] Implementation of the methods and individual steps thereof disclosed herein can be done manually and / or with the aid of electronic equipment. Although the processes have been described with reference to particular embodiments, a person of ordinary skill in the art will readily appreciate that other ways of performing the acts associated with the methods can be used. For example, the order of various steps can be changed, unless otherwise described, without departing from the scope or spirit of the methods. In addition, some of the individual steps can be combined, omitted, or further subdivided into additional steps.

[0190] All patents, publications, and references cited herein are fully incorporated by reference. In the event of an inconsistency between the present disclosure and the incorporated patents, publications, and references, the present disclosure shall dominate.

[0191] Embodiments contemplated in view of the foregoing include those described in the following numbered paragraphs.

[0192] 1. A controlled release formulation of a vitamin D compound comprising one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3, the formulation comprising a matrix that releasably binds and controllably releases the vitamin D compound, the matrix comprising a cellulose derivative.

[0193] 2. A stabilized formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject who ingests the formulation, the formulation comprising a mixture of:

[0194] one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3;

[0195] and an effective amount of a stabilizing agent, optionally a cellulose compound, to maintain a difference of less than 30% between the amount of vitamin D compound released at any given time point after four hours during an in vitro dissolution test and the amount released at the same dissolution time point during an in vitro dissolution conducted prior to exposing the formulation to the storage conditions, after two months exposure to storage conditions of 25°C and 60% relative humidity.

[0196] 3. A stabilized formulation for controlled release of a vitamin D compound, the formulation comprising a mixture of:

[0197] one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3;

[0198] a wax matrix; and

[0199] a stabilizer, optionally a cellulose compound.

[0200] 4. A stabilized formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation, the formulation comprising a mixture of:

[0201] one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3;

[0202] a wax matrix; and

[0203] a stabilizer, optionally a cellulose stabilizer.

[0204] 5. A stabilized formulation for controlled release of a vitamin D, the formulation comprising a mixture of:

[0205] one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3;

[0206] a wax matrix; and

[0207] an effective amount of a stabilizer, optionally a cellulose compound, to maintain a difference of less than 30% between the amount of vitamin D compound released at any given time point after four hours during an in vitro dissolution test and the amount released at the same dissolution time point during an in vitro dissolution conducted prior to exposing the formulation to storage conditions of two months exposure to 25°C and 60% relative humidity.

[0208] 6. A stabilized formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation, the formulation comprising a mixture of:

[0209] an active agent-loaded wax matrix comprising one or both of 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3; and

[0210] a cellulose stabilizer;

[0211] wherein the amount of 25-hydroxyvitamin D released during in vitro dissolution after a two month storage period at 25 °C and 60% relative humidity of the formulation varies by 30% or less at any given dissolution time point in the absence of the cellulose stabilizer compared to the amount released at the same dissolution time point during in vitro dissolution performed prior to exposing the formulation to the storage conditions.

[0212] 7. In a formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation, the improvement comprising admixing a cellulose stabilizer into the formulation.

[0213] 8. In a formulation for controlled release of a vitamin D compound in the gastrointestinal tract of a subject ingesting the formulation, the improvement comprising admixing into the formulation an effective amount of a cellulose compound to maintain a variation of less than 30% at any given dissolution time point after four hours of the amount of 25-hydroxyvitamin D released during in vitro dissolution after exposure to storage conditions of 25 °C and 60% relative humidity for at least one month compared to the amount released at the same dissolution time point during in vitro dissolution performed prior to exposing the formulation to the storage conditions.

[0214] 9. The formulation of any of the preceding paragraphs, wherein the cellulose compound or cellulose stabilizer comprises a cellulose ether.

[0215] 10. The formulation of paragraph 9, wherein the cellulose ether is selected from the group consisting of methyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl methyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose.

[0216] 11. The formulation of paragraph 9, wherein the cellulose compound or cellulose stabilizer is hydroxypropyl methyl cellulose.

[0217] 12. The formulation of any of the preceding paragraphs, wherein the amount of 25-hydroxyvitamin D released during in vitro dissolution after exposure to storage conditions of 25 °C and 60% relative humidity for 2 months varies by 30% or less at any given dissolution time point after four hours compared to the amount released at the same dissolution time point during in vitro dissolution performed prior to exposing the formulation to the storage conditions.

[0218] 13. The formulation of any of the preceding paragraphs, wherein the amount of 25-hydroxyvitamin D released during in vitro dissolution after exposure to storage conditions of 40 °C and 75% relative humidity for one month varies by 30% or less at any given dissolution time point after four hours compared to the amount released at the same dissolution time point during in vitro dissolution performed prior to exposing the formulation to the storage conditions.

[0219] 14. The formulation of any of the preceding paragraphs, wherein the matrix comprises a wax matrix comprising a controlled release agent, an emulsifying agent, and an absorption enhancer.

[0220] 15. The formulation of paragraph 14, wherein the controlled release agent comprises paraffin wax.

[0221] 16. The formulation of paragraph 14 or 15, wherein the emulsifying agent has an HLB value of less than 7.

[0222] 17. The formulation of paragraph 16, wherein the emulsifying agent comprises glyceryl monostearate.

[0223] 18. The formulation of any of paragraphs 14 to 17, wherein the absorption enhancer has an HLB value in the range of about 13 to about 18.

[0224] 19. The formulation of paragraph 18, wherein the absorption enhancer is a mixture of lauroyl polyoxylglycerols and lauroyl macrogolglycerides.

[0225] 20. The formulation of any of the preceding paragraphs, wherein the vitamin D compound comprises 25-hydroxyvitamin D3.

[0226] 21. The formulation of any of the preceding paragraphs, further comprising an oily vehicle.

[0227] 22. The formulation of paragraph 21, wherein the oily vehicle comprises mineral oil.

[0228] 23. The formulation of paragraph 22, wherein the formulation comprises about 20 wt% paraffin wax, about 20 wt% to about 25 wt% glyceryl monostearate, about 10 wt% a mixture of lauroyl polyoxylglycerols and lauroyl macrogolglycerides, about 30 wt% to about 35 wt% mineral oil, and about 10 wt% to about 15 wt% hydroxypropyl methylcellulose.

[0229] 24. The formulation of any of the preceding paragraphs, wherein the formulation comprises glyceryl monostearate.

[0230] 25. The formulation of any of the preceding paragraphs, wherein the formulation comprises one or more polyoxylated glycerides.

[0231] 26. A sustained release dosage form in the form of a capsule, tablet, sachet, dragee, or suppository, comprising the formulation of any of the preceding paragraphs.

[0232] 27. The dosage form of paragraph 26, comprising a capsule or a tablet.

[0233] 28. The dosage form of paragraph 27, comprising a capsule.

[0234] 29. The dosage form of paragraph 26, comprising an oral capsule, tablet, sachet, dragee.

[0235] 30. The stabilized dosage form of any of the preceding paragraphs, characterized by providing a dissolution profile of release of the vitamin D compound:

[0236] less than 30% at 2 hours;

[0237] greater than 45% at 6 hours; and

[0238] greater than 80% at 12 hours.

[0239] 31. The stabilized dosage form of paragraph 26, wherein the release of the vitamin D compound is less than 60% at 6 hours.

[0240] 32. A stabilized sustained release oral dosage form comprising a vitamin D compound, characterized by providing an in vitro dissolution profile of release of the vitamin D compound:

[0241] less than 30% at 100 to 140 minutes;

[0242] greater than 45% at 5 to 7 hours; and

[0243] greater than 80% at 11 to 13 hours.

[0244] 33. The dosage form of paragraph 32, wherein the release of the vitamin D compound is

[0245] less than 30% at 2 hours;

[0246] greater than 45% at 6 hours; and

[0247] greater than 80% at 12 hours.

[0248] 34. The dosage form of paragraph 32 or 33, wherein the release of the vitamin D compound is less than 60% at 5 to 7 hours.

[0249] 35. The dosage form of paragraph 34, wherein the release of the vitamin D compound is less than 60% at 6 hours.

[0250] 36. A stabilized sustained release oral dosage form comprising a vitamin D compound, characterized by providing an in vitro dissolution profile of release of the vitamin D compound:

[0251] about 20% to about 40% at 2 hours;

[0252] at least 35% at 6 hours; and

[0253] at least 70% at 12 hours.

[0254] 37. The dosage form of paragraph 36, wherein the release of the vitamin D compound is about 25% to about 35% at 2 hours;

[0255] at least 40% at 6 hours; and

[0256] at least 75% at 12 hours.

[0257] 38. The dosage form of paragraph 36 or 37, wherein the release of the vitamin D compound is 75% or less at 6 hours.

[0258] 39. The dosage form of paragraph 38, wherein the release of the vitamin D compound is 65% or less at 6 hours.

[0259] 40. The dosage form of paragraph 39, wherein the release of the vitamin D compound is 60% or less at 6 hours.

[0260] 41. A stabilized sustained release dosage form comprising a vitamin D compound characterized by a t max .

[0261] 42. The dosage form of paragraph 41, wherein the t max is at least 8 hours.

[0262] 43. The dosage form of paragraph 42, wherein the t max is at least 12 hours.

[0263] 44. The dosage form of paragraph 43, wherein the t max is at least 18 hours.

[0264] 45. The dosage form of paragraph 44, wherein the t max is at least 20 hours.

[0265] 46. The dosage form of paragraph 45, wherein the t max is at least 24 hours.

[0266] 47. The dosage form of paragraph 46, wherein the t max is at least 28 hours.

[0267] 48. The dosage form of paragraph 41, wherein the t max ranges from 4 to 96 hours.

[0268] 49. The dosage form of paragraph 48, wherein the t max ranges from 18 to 30 hours.

[0269] 50. The dosage form of paragraph 49, wherein the t max is in the range of 13 to 28 hours.

[0270] 51. The dosage form of paragraph 50, wherein the t max is about 28 hours.

[0271] 52. A stabilized, sustained release dosage form comprising a 25-hydroxyvitamin D compound, the dosage form characterized as providing a baseline-adjusted C max .

[0272] 53. A method of administering a stabilized, sustained release dosage form comprising a 25-hydroxyvitamin D compound to a human patient, comprising administering an effective amount of the dosage form to the patient to provide a baseline-adjusted C max .

[0273] 54. The method of paragraph 53, comprising administering an effective amount of the dosage form to provide a baseline-adjusted C max .

[0274] 55. A method of administering a stabilized, sustained release dosage form comprising a 25-hydroxyvitamin D compound to a human patient, comprising administering an effective amount of the dosage form to the patient to provide a baseline-adjusted AUC 0-inf .

[0275] 56. The method of paragraph 55, comprising administering an effective amount of the dosage form to the patient to provide a baseline-adjusted AUC 0-inf .

[0276] 57. A method of vitamin D supplementation, comprising administering to a subject in need thereof a formulation or dosage form of any of the preceding paragraphs.

[0277] 58. A method of treating or preventing a vitamin D responsive disease in a subject, comprising administering to the subject a formulation or dosage form of any of the preceding paragraphs.

[0278] 59. The method of paragraph 58, wherein the disease is selected from cancer (e.g., breast cancer, lung cancer, skin cancer, melanoma, colon cancer, colorectal cancer, rectal cancer, prostate cancer, and bone cancer); an autoimmune disease, e.g., Type I diabetes mellitus, multiple sclerosis, rheumatoid arthritis, polymyositis, dermatomyositis, scleroderma, fibrosis, Graves' disease, Hashimoto's disease, acute or chronic transplant rejection, acute or chronic graft-versus-host disease, inflammatory bowel disease, Crohn's disease, systemic lupus erythematosus, Sjogren's syndrome, eczema, and psoriasis; dermatitis, including atopic dermatitis, contact dermatitis, allergic dermatitis, and / or chronic dermatitis; an inflammatory disease, e.g., asthma, chronic obstructive pulmonary disease, polycystic kidney disease, polycystic ovary syndrome, pancreatitis, nephritis, hepatitis, and / or infection; hypertension; a cardiovascular disease, e.g., a subject with atherosclerosis, arteriosclerosis, coronary artery disease, cerebrovascular disease, peripheral vascular disease, myocardial infarction, myocardial ischemia, cerebral ischemia, stroke, congestive heart failure, cardiomyopathy; obesity or other body weight abnormalities; a lipid disorder (e.g., hyperlipidemia; dyslipidemia, including associated diabetic dyslipidemia and mixed dyslipidemia; hypoalphalipoproteinemia; hypertriglyceridemia; hypercholesterolemia; and low HDL (high density lipoprotein)); a metabolic disorder (e.g., metabolic syndrome; Type II diabetes mellitus; Type I diabetes mellitus; hyperinsulinemia; impaired glucose tolerance; insulin resistance; diabetic complications, including neuropathy, nephropathy, osteoporosis, retinopathy, diabetic foot ulcer, and cataracts); and / or thrombosis.

[0279] 60. The method of paragraph 59, wherein the disease is selected from (i) in the parathyroid gland— hypoparathyroidism, pseudohypoparathyroidism, secondary hyperparathyroidism; (ii) in the pancreas— diabetes mellitus; (iii) in the thyroid— medullary carcinoma; (iv) in the skin— psoriasis, wound healing; (v) in the lung— sarcoidosis and tuberculosis; (vi) in the kidney— chronic kidney disease, hypophosphatemic VDRR, vitamin D dependent rickets; (vii) in the bone— anticonvulsant therapy, poor bone fibrogenesis, osteitis fibrosa cystica, osteomalacia, osteoporosis, osteopenia, osteosclerosis, renal osteodystrophy, rickets; (viii) in the intestine— glucocorticoid antagonism, idiopathic hypercalcemia, malabsorption syndrome, steatorrhea, tropical sprue; and (ix) autoimmune disorders.

[0280] 61. The method of paragraph 60, wherein the disease is selected from cancer, a skin disorder (e.g., psoriasis), a parathyroid disorder (e.g., hyperparathyroidism and secondary hyperparathyroidism), a bone disorder (e.g., osteoporosis), and an autoimmune disorder.

[0281] 62. The method of paragraph 61, wherein the disease is secondary hyperparathyroidism.

[0282] 63. The method of paragraph 62, wherein the subject has chronic kidney disease (CKD).

[0283] 64. The method of paragraph 63, wherein the CKD is stage 3 or stage 4.

[0284] 65. The method of paragraph 64, wherein the patient is vitamin D deficient.

[0285] 66. The method of any of the preceding paragraphs, wherein the patient is a human.

[0286] 67. The method of paragraph 66, wherein the human is an adult.

[0287] 68. A composition substantially as described herein.

Claims

1. A storage stable formulation for sustained release of a vitamin D compound, the formulation comprising: a lipophilic matrix, 25-hydroxyvitamin D3, and a stabilizer, the stabilizer being hydroxypropyl methylcellulose, wherein the stabilizer is effective to maintain a difference of less than 30% between the amount of vitamin D compound released at any given time point after four hours during in vitro dissolution testing following two months of exposure to storage conditions of 25°C and 60% relative humidity, and the amount released at the same dissolution time point during in vitro dissolution conducted prior to exposing the formulation to the storage conditions, wherein the formulation comprises 20 to 30 wt% paraffin, 20 to 25 wt% glyceryl monostearate, 10 wt% of a mixture of lauroyl macrogolglycerides and lauroyl polyoxylglycerides, 30 to 35 wt% mineral oil, and 10 to 15 wt% hydroxypropyl methylcellulose.

2. A sustained release dosage form in the form of a capsule, tablet, sachet, dragee, or suppository comprising the formulation of claim 1.

3. The sustained release dosage form of claim 2, wherein the dosage form comprises 5 to 80 μg of 25-hydroxyvitamin D3.

4. The sustained release dosage form of claim 3, wherein the dosage form comprises 30 or 60 μg of 25-hydroxyvitamin D3.

5. Use of the storage stable formulation for sustained release of claim 1 or the sustained release dosage form of any one of claims 2-4 in the manufacture of a medicament for treating a patient having a vitamin D responsive disease.

6. The use of claim 5, wherein the vitamin D responsive disease is selected from the group consisting of a cancer, a skin disorder, a parathyroid disorder, a bone disorder, and an autoimmune disorder.

7. The use of claim 5, wherein the vitamin D responsive disease is selected from the group consisting of psoriasis, primary hyperparathyroidism, secondary hyperparathyroidism, and osteoporosis.

8. The use of claim 5, wherein the vitamin D responsive disease is secondary hyperparathyroidism.

9. The use of any one of claims 5-8, wherein the patient has chronic kidney disease (CKD).

10. The use of any one of claims 5-8, wherein the patient has chronic kidney disease that is stage 3 or stage 4.

11. The use of any one of claims 5-8, wherein the patient has chronic kidney disease that is stage 3 or stage 4 and is vitamin D insufficient.

12. The use of any one of claims 5-8, wherein the patient has chronic kidney disease that is stage 3 or stage 4 and is vitamin D deficient.

13. The use of claim 5, wherein the vitamin D responsive disease is hyperparathyroidism secondary to chronic kidney disease.

14. The use of claim 13, wherein the vitamin D responsive disease is hyperparathyroidism secondary to chronic kidney disease stage 3 or stage 4.

15. The use of any one of claims 5-8, wherein the patient is a human. ​ 16. The use of any one of claims 5-8, wherein the patient is an adult.

17. The use of any one of claims 5-8, wherein 5 μg to 80 μg of 25-hydroxyvitamin D3 is administered per day.

18. The use of any one of claims 5-8, wherein 30 μg or 60 μg of 25-hydroxyvitamin D3 is administered per day.

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