US20190134059A1

US20190134059A1 - Fulvestrant formulations and methods of their use

Info

Publication number: US20190134059A1
Application number: US16/099,265
Authority: US
Inventors: Feng-Jing Chen; Steven L. Krill; Rama Abu Shmeis
Original assignee: Eagle Pharmaceuticals Inc
Current assignee: Eagle Pharmaceuticals Inc
Priority date: 2016-05-06
Filing date: 2017-05-05
Publication date: 2019-05-09
Also published as: MA44862A; US20210346396A1; AU2017261321B2; KR20190005183A; EP3452011A1; BR112018072768A2; ZA201807031B; MX2018013414A; JP2023109959A; RU2018142068A; CA3022834A1; CO2018013257A2; MX2022013199A; CN109310621A; IL285928A; RU2018142068A3; WO2017193048A1; UA125514C2; JP2021169510A; AU2017261321A1

Abstract

The disclosure is directed to fulvestrant formulations including suspensions of fulvestrant particles suitable for injection. The formulations can comprise fulvestrant particles having an LD Dv(10) less than about 3 microns, for example, between about 1 micron to about 3 microns, an LD Dv(50) less than about 35 microns, for example, between about 2 microns and about 35 microns, and an LD Dv(90) less than about 120 microns, for example, between about 4 microns and about 120 microns. The formulations can comprise fulvestrant particles having a CE Dv(90) less than about 200 microns, for example, between about 10 microns and about 200 microns, a CE Dv(50) less than about 60 microns, for example, between about 5 microns and about 60 microns, and a CE Dv(10) less than about 25 microns, for example, between about 1 micron and about 25 microns.

Description

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No. 62/332,842, filed May 6, 2016, and U.S. Provisional Application No. 62/420,555, filed Nov. 10, 2016, the entireties of which are incorporated by reference herein.

FIELD

The disclosure is directed to fulvestrant-containing formulations and methods of their use in the treatment of disease.

BACKGROUND

Fulvestrant, or 7-(9-(4,4,5,5,5-pentafluoropentylsulfinyl)nonyl)estra-1,3,5(10)-triene-3,17-diol, has the structure of formula (1):
Fulvestrant is a selective estrogen receptor degrader (SERD) indicated for the treatment of hormone receptor positive metastatic breast cancer in postmenopausal women with disease progression following anti-estrogen therapy.
As with other steroidal-like compounds, fulvestrant has physical properties which make preparing fulvestrant pharmaceutical compositions difficult. Fulvestrant is a particularly lipophilic molecule, even when compared with other steroidal compounds, and its aqueous solubility is extremely low.
Due to the poor solubility and oral bioavailability of fulvestrant, the drug is currently administered via intramuscular injection of an oil-based fulvestrant formulation. The current commercial formulation of fulvestrant, FASLODEX™, is dosed at 500 mg and requires that two 5 mL injections of a 50 mg/mL fulvestrant formulation be administered intramuscularly. Each 5 mL injection contains 10% w/v alcohol, 10% w/v benzyl alcohol, and 15% w/v benzyl benzoate as co-solvents and made up to 100% w/v with castor oil as a further co-solvent and release rate modifier. Administration of the formulation is slow (1-2 minutes per injection) and painful, due to the viscous oil-based vehicle used to solubilize fulvestrant. A warning has been added to the FASLODEX™ label concerning painful injections, sciatica, neuropathic pain, and peripheral neuropathy.
It has been previously reported (U.S. Pat. No. 6,774,122 to AstraZeneca) that intra-muscular injections of fulvestrant in the form of an aqueous suspension were not suitable for use. Those suspensions resulted in extensive local tissue irritation at the injection site as well as a poor release profile due to the presence of fulvestrant in the form of solid particles. Furthermore, the fulvestrant release rate was reported as not clinically significant.
There is a need for fulvestrant formulations with improved dosing properties. The disclosure is directed to these and other important needs.

SUMMARY

The present disclosure provides formulations comprising fulvestrant particles. The disclosure also provides fulvestrant suspensions, preferably those having a fulvestrant concentration of equal to or greater than about 50 mg/mL. The disclosure also provides formulations comprising fulvestrant particles and a non-oil vehicle. Some aspects of the disclosure are directed to pharmaceutical compositions comprising fulvestrant particles having an LD Dv(90) greater than or equal to about 7 microns. Further aspects of the disclosure are directed to pharmaceutical compositions comprising fulvestrant particles having a CE Dv(90) less than about 200 microns, for example, between about 10 microns and about 200 microns, a CE Dv(50) less than about 60 microns, for example, between about 5 microns and about 60 microns, and a CE Dv(10) less than 25 microns, for example, between about 1 microns and about 25 microns. Other aspects of the disclosure are directed to pharmaceutical compositions comprising fulvestrant at a concentration of about 100 mg/mL, whereupon administration to a subject, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of the pharmaceutical compositions of the disclosure is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of a reference listed fulvestrant product. Other aspects of the disclosure are directed to fulvestrant formulations having a concentration of about 100 mg/mL and particular pharmacokinetic profiles. In other aspects, the disclosure is directed to pharmaceutical compositions comprising fulvestrant particles, wherein the fulvestrant concentration is about 40 to 125 mg/mL.
Methods of making and using the products described herein are also described.

BRIEF DESCRIPTION OF THE DRAWINGS

The summary, as well as the following detailed description, is further understood when read in conjunction with the appended drawings. For the purpose of illustrating the disclosure, there are shown in the drawings exemplary embodiments of the disclosure; however, the disclosure is not limited to the specific methods, compositions, and devices disclosed. In the drawings:

FIG. 1A depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 1B depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 2A depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 2B depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 2C depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 3 depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 4 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 5 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 6 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 7 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 8 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 9 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 10 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 11 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 12 depicts aspects of exemplary methods of preparation for fulvestrant suspensions of the present disclosure;

FIG. 13 depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines;

FIG. 14 depicts pharmacokinetic data for administration of commercial fulvestrant formulations (FASLODEX™) and some exemplary fulvestrant formulations of the present disclosure to canines; and

FIG. 15 depicts schematic representations of aspects of methods of preparation of some exemplary fulvestrant formulations of the present disclosure.

All callouts and annotations in the Figures are hereby incorporated into this description as if fully set forth herein.

DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

The present disclosure may be understood more readily by reference to the following detailed description taken in connection with the accompanying figures and examples, which form a part of this disclosure. It is to be understood that this disclosure is not limited to the specific devices, methods, applications, conditions or parameters described and/or shown herein, and that the terminology used herein is for the purpose of describing particular embodiments by way of example only and is not intended to be limiting of the claimed disclosure.
As used in the specification including the appended claims, the singular forms “a,” “an,” and “the” include the plural, and reference to a particular numerical value includes at least that particular value, unless the context clearly dictates otherwise.
When a range of values is expressed, another embodiment includes from the one particular value and/or to the other particular value. All ranges are inclusive and combinable. Further, reference to values stated in ranges include each and every value within that range. When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. The term “about” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass reasonable variations of the value, such as, for example, ±10% from the specified value. For example, the phrase “about 50%” can include ±10% of 50, or from 45% to 55%.
It is to be appreciated that certain features of the disclosure which are, for clarity, described herein in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any subcombination.

Terms

As used herein, whether by itself or in conjunction with another term or terms, it should be understood that the phrases “method of treating” and “method of treatment” may be used interchangeably with the phrase “for use in the treatment of” a particular disease.
As used herein, whether by itself or in conjunction with another term or terms, “pharmaceutically acceptable” indicates that the designated entity such as, for example, e.g., a pharmaceutically acceptable excipient is generally chemically and/or physically compatible with other ingredients in a formulation, and/or is generally physiologically compatible with the recipient thereof.
As used herein, “pharmaceutical composition” refers to a formulation as described herein that includes one or more pharmaceutically acceptable excipients, that is suitable for administration to a subject. It should be understood that the term “pharmaceutical composition” encompasses (a) suspensions and (b) suspensions which have been dried such that one or more solvents have been removed partially or completely, either by evaporation or sublimation, including, but not limited to, lyophilized cakes.
As used herein, whether by themselves or in conjunction with another term or terms, “subject(s),” “individual(s),” and “patient(s)”, refer to mammals, including humans. The term human(s) refers to and includes, a human child, adolescent, or adult.
As used herein, whether by themselves or in conjunction with another term or terms, “treats,” “treating,” “treated,” and “treatment,” refer to and include ameliorative, palliative, and/or curative uses and results, or any combination thereof. In other embodiments, the methods described herein can be used prophylactically. It should be understood that “prophylaxis” or a prophylactic use or result do not refer to nor require absolute or total prevention (i.e., a 100% preventative or protective use or result). As used herein, prophylaxis or a prophylactic use or result refer to uses and results in which administration of a compound or formulation diminishes or reduces the severity of a particular condition, symptom, disorder, or disease described herein; diminishes or reduces the likelihood of experiencing a particular condition, symptom, disorder, or disease described herein; or delays the onset or relapse (reoccurrence) of a particular condition, symptom, disorder, or disease described herein; or any combination of the foregoing.
As used herein, whether used alone or in conjunction with another term or terms, “therapeutic” and “therapeutically effective amount”, refer to an amount of a compound or formulation that (a) treats a particular condition, symptom, disorder, or disease described herein; (b) attenuates, ameliorates or eliminates one or more symptoms of a particular condition, disorder, or disease described herein; (c) delays the onset or relapse (reoccurrence) of a particular condition, symptom, disorder, or disease described herein. It should be understood that the terms “therapeutic” and “therapeutically effective” encompass any one of the aforementioned effects (a)-(c), either alone or in combination with any of the others (a)-(c).
As used herein, whether used alone or in conjunction with another term or terms, “therapeutic agent” refers to any substance included in a formulation that is useful in the treatment of a disease, condition, or disorder or comorbidity (i.e., a disease, condition, or disorder that exists simultaneously with breast cancer) and is not fulvestrant.
As used herein, whether used alone or in conjunction with another term or terms, “suspension” refers to solid particles dispersed in a liquid vehicle.
As used herein, whether used alone or in conjunction with another term or terms, “formulation” refers to a mixture of components. The term “formulation” encompasses pharmaceutical compositions, and suspensions, as well as suspensions that have been dried such that one or more solvents have been removed partially or completely (e.g., lyophilized cakes).
As used herein “Dv(10)”, “Dv(50)” and “Dv(90)” are defined as the volume weighted particle diameters where a cumulative 10%, 50% or 90% v/v of the particles have an equal or smaller diameter, respectively, when measured. For example, if a particle population has a Dv(50) of about 25 microns, 50% of the particles in volume have a diameter of less than or equal to about 25 microns.
As used herein, Dn(10)”, “Dn(50)” and “Dn(90)” are defined as the number weighted particle diameters where a cumulative 10%, 50% or 90% of the particles have an equal or smaller diameter, respectively, when measured. For example, if a particle population has a Dn(50) of about 25 microns, 50% of the particles in number have a diameter of less than or equal to about 25 microns.
Particle size and particle size distributions can be determined by measurement via laser diffraction. Particle size analysis by laser diffraction methods is known in the art and is explained more fully by ISO 13320:2009(E), “Particle size analysis—Laser diffraction methods,” International Organization for Standardization which is incorporated by reference herein in its entirety for all purposes. Particle sizes determined by laser diffraction are represented as the diameter of a sphere having equivalent volume to the particle volume as determined by Mie theory of light scattering. Tables 1-7 and 23-27 and FIGS. 4-12 provide laser diffraction particle size and particle size distribution (“PSD”) data for some exemplary embodiments of the present invention, with measurements taken during methods of preparation, on the day of formulation (“Day 0”), and at various later dates after formulation, as indicated. Measurements were taken “as is” and “sonicated.” Data for “sonicated” samples indicates that the measurement sample was subjected to sonication to disperse agglomerates and provide stable repeat measurements, as more fully described in ISO 13320:2009(E). Values measured via laser diffraction are indicated as such in the Figures and Tables, or are referred to herein by “laser diffraction Dv(##)”, “LD Dv(##)”, “laser diffraction diameter”, or “LD diameter.”
Particle size and particle size distributions can also be determined by microscopy image capture and analysis. Microscopy image capture and analysis captures a two dimensional (2D) image of a 3D particle and calculates various size and shape parameters from the 2D image. Particle sizes determined by microscopy image capture and analysis are represented as the diameter of a circle with the equivalent area as the 2D image of the particle, referred to herein as a circle equivalent or “CE” diameter. Particle size analysis by microscopy image capture and analysis is known in the art and is explained more fully by ISO 13322-1:2014, “Particle size analysis—Image analysis methods—Part 1: Static image analysis methods,” International Organization for Standardization, which is incorporated by reference herein in its entirety for all purposes. Values measured by microscopy image capture and analysis are referred to herein by “circle equivalent diameter,” “CE diameter,” “circle equivalent Dv(##),” “CE Dv(##)”, or “CE Dn(##)”. Tables 41-50 provide microscopy image capture and analysis particle size and particle size distribution data for some exemplary embodiments of the present invention, with measurement samples taken during methods of preparation, after an initial suspension is formed, or after lyophilization and reconstitution, as indicated.

A. Formulations.

Suspensions Comprising Fulvestrant Particles and a Vehicle

In particular embodiments, the invention is directed to suspensions comprising fulvestrant particles and a vehicle. The fulvestrant particles may have different particle size distributions as described more fully elsewhere herein. As used herein, a “vehicle” is a suspending medium, preferably a pharmaceutically acceptable suspending medium. In certain embodiments, the vehicle is a non-oil vehicle. As used herein, “oils” are non-polar substances that have no or low miscibility with water. Castor oil is an example of an oil. In other embodiments of the invention, the vehicle comprises water, i.e., is aqueous. As used herein, an “aqueous” vehicle is a vehicle that comprises at least about 50% w/w water. In some embodiments, the aqueous vehicle comprises at least about 60% w/w, at least about 70% w/w, at least about 80% w/w, at least about 85% w/w, at least about 90% w/w, at least about 95% w/w, at least about 96% w/w, at least about 97% w/w, at least about 98% w/w, or at least about 99% w/w water. In certain embodiments of the invention, the vehicle is water. In yet other embodiments of the invention, the vehicle is a non-aqueous medium. In some embodiments, a vehicle comprises a single suspending medium. In other embodiments, a vehicle comprises a mixture of two or more suspending mediums, which may be aqueous or non-aqueous. In still other embodiments of the invention, the vehicle comprises both water and a non-aqueous solvent. In particular embodiments of the invention, the suspension is substantially oil-free. As used herein, a “substantially oil-free” suspension is a suspension comprising a vehicle that comprises at most about 10% w/w oil. In some preferred embodiments, a substantially oil-free suspension comprises a vehicle that comprises less than about 5% w/w oil, less than about 2% w/w oil, less than about 1% w/w oil, less than about 0.5% w/w oil, less than about 0.1% w/w oil, or comprises a vehicle that is free of oil.
Fulvestrant suspensions of the disclosure can have fulvestrant present at a concentration of about 40 mg/mL to about 125 mg/mL in a vehicle. The fulvestrant present in the fulvestrant suspensions may have different particle size distributions as described more fully elsewhere herein. In particular embodiments of the invention, fulvestrant is present at a concentration equal to or greater than about 40 mg/mL. In further embodiments, fulvestrant is present at a concentration of about 40 to about 75 mg/mL. In other embodiments, fulvestrant is present at a concentration of about 75 mg/mL to about 125 mg/mL. In still further embodiments, fulvestrant is present at a concentration of about 40 mg/mL, about 45 mg/mL, about 50 mg/mL, about 55 mg/mL, about 60 mg/mL, about 65 mg/mL, about 70 mg/mL, or about 75 mg/mL.
In certain embodiments, fulvestrant is present in the suspension at a concentration equal to or greater than about 75 mg/mL. In further embodiments, fulvestrant is present in the suspension at a concentration of about 75 to about 125 mg/mL. In particular embodiments, fulvestrant is present in the suspension at a concentration of about 80 mg/mL, about 85 mg/mL, about 90 mg/mL, about 95 mg/mL, about 100 mg/mL, about 105 mg/mL, about 110 mg/mL, about 115 mg/mL, about 120 mg/mL, or about 125 mg/mL. In other embodiments, fulvestrant is present in the suspension at a concentration of about 75 mg/mL to about 95 mg/mL, about 80 mg/mL to about 100 mg/mL, about 90 mg/mL to about 110 mg/ml, about 95 mg/mL to about 105 mg/mL, about 95 mg/mL to about 115 mg/mL, about 100 mg/mL to about 110 mg/mL, about 110 mg/mL to about 125 mg/mL, including all ranges and subranges there between.

Pharmaceutical Compositions Comprising Fulvestrant

Other embodiments of the disclosure include pharmaceutical compositions comprising fulvestrant. These pharmaceutical compositions may be prepared by combining fulvestrant, as described herein, with one or more additional excipients, preferably pharmaceutically acceptable excipients.
In certain embodiments, the pharmaceutical compositions may further comprise a stabilizer, or one or more stabilizers, or two or more stabilizers. In still further embodiments of the invention, the stabilizer is selected from the group consisting of surfactants, polymers, cross-linked polymers, buffering agents, electrolytes, and non-electrolytes. In yet further embodiments of the invention, the pharmaceutical composition comprises a combination of two or more stabilizers selected from the group consisting of surfactants, polymers, cross-linked polymers, buffering agents, electrolytes, and non-electrolytes.
In certain embodiments of the invention, the pharmaceutical compositions comprising fulvestrant comprise about 0.2 mg/mL to about 75 mg/mL of one or more stabilizers, and all ranges and subranges therebetween. In particular embodiments of the invention, the pharmaceutical composition comprises about 0.2 to 0.7 mg/mL, 0.5 to 1 mg/mL, 1 to 5 mg/mL, 2 to 8 mg/mL, 5 to 6 mg/mL, 5 to 10 mg/mL, 8 to 12 mg/mL, 10 to 15 mg/mL, 15 to 20 mg/mL, 20 to 30 mg/mL, 30 to 40 mg/mL, 40 to 50 mg/mL, 45 to 55 mg/mL, 50 to 60 mg/mL, or 60 to 75 mg/mL of one or more stabilizers, and all ranges and subranges there between. In further embodiments of the invention, the pharmaceutical composition comprises about 0.2 mg/mL, 0.5 mg/mL, 1 mg/mL, 2 mg/mL, 3 mg/mL, 4 mg/mL, 5 mg/mL, 5.5 mg/mL, 6 mg/mL, 7 mg/mL, 8 mg/mL, 9 mg/mL, 10 mg/mL, 12 mg/mL, 15 mg/mL, 17 mg/mL, 20 mg/mL, 25 mg/mL, 30 mg/mL, 35 mg/mL, 40 mg/mL, 45 mg/mL, 50 mg/mL, 55 mg/mL, 60 mg/mL, 65 mg/mL, 70 mg/mL, or about 75 mg/mL of one or more stabilizers.
In yet further embodiments of the invention, the stabilizer is a surfactant. For example, the stabilizer can be, but is not limited to, polyethylene oxide (PEO), a PEO derivative, polysorbate 80, polysorbate 20, poloxamer 188 (including, but not limited to, PLURONIC® F-68 poloxamer sold by BASF Corp. (Wyandotte, Mich., USA)), poloxamer 124 (including, but not limited to, PLURONIC® L44 poloxamer sold by BASF Corp. (Wyandotte, Mich., USA)), poloxamer 407 (including, but not limited to, PLURONIC® F127 poloxamer sold by BASF Corp. (Wyandotte, Mich., USA)), polyethoxylated vegetable oils, polyethoxylated castor oil (including but not limited to KOLLIPHOR® EL, formerly known as CREMOPHOR® EL sold by BASF Corp. (Wyandotte, Mich., USA)), sorbitan palmitate (including, but not limited to, SPAN™ 40 sold by Croda International Plc), lecithin, poly(vinyl alcohol) (“PVA”), human serum albumin, and mixtures thereof.
In particular embodiments of the invention, the stabilizer is a polymer. For example, the stabilizer can be, but is not limited to, a polyvinylpyrrolidone (“PVP”) (such as, but not limited to povidone K12, povidone K17, PLASDONE™ C-12 povidone, PLASDONE™ C-17 povidone, PLASDONE™ C-30 povidone, and mixtures thereof), polyethylene glycol 3350, and mixtures thereof.
In other embodiments of the invention, the stabilizer is an electrolyte, i.e., a salt that dissociates into anions and cations in aqueous solution. For example, the stabilizer can be, but is not limited to, sodium chloride, calcium chloride, and mixtures thereof.
In still other embodiments of the invention, the stabilizer is a non-electrolyte, i.e., is non-ionic. For example, the stabilizer can be, but is not limited to, dextrose, glycerol (also referred to as glycerin), mannitol, or mixtures thereof.
In other embodiments of the invention, the stabilizer is a cross-linked polymer. For example, the stabilizer can be, but is not limited to, carboxymethylcellulose sodium (CMC). In some embodiments of the invention, the stabilizer is CMC 7LF, CMC 7MF, CMC 7HF, or mixtures thereof.
In other embodiments of the invention, the stabilizer is a buffering agent, for example, NaH₂PO₄.H₂O, NaH₂PO₄.2H₂O, anhydrous NaH₂PO₄, sodium citrate, citric acid, Tris, sodium hydroxide, HCl, or mixtures thereof.
In further embodiments of the invention, combinations of non-electrolyte stabilizers and electrolyte stabilizers may be used. In some embodiments, the combination of stabilizers may comprise two or more non-electrolyte stabilizers. In other embodiments, the combination of stabilizers may comprise two or more electrolyte stabilizers. In further embodiments, the combination of stabilizers may comprise one or more non-electrolyte stabilizers and one or more electrolyte stabilizers. In yet further embodiments, the combination of stabilizers may comprise two or more of mannitol, dextrose, and sodium chloride.
In certain embodiments of the invention, combinations of surfactant stabilizers and polymer stabilizers may be used. In some embodiments, the combination of stabilizers may comprise two or more surfactant stabilizers. In other embodiments, the combination of stabilizers may comprise two or more polymer stabilizers. In further embodiments, the combination of stabilizers may comprise one or more surfactant stabilizers and one or more polymer stabilizers. In yet further embodiments, the combination of stabilizers may comprise two or more of polysorbate 80, polysorbate 20, and poloxamer 188. In still further embodiments, the combination of stabilizers may comprise one or more of polysorbate 80, polysorbate 20, and poloxamer 188 and one or more of povidone K12, povidone K17, PLASDONE™ C-12 povidone, PLASDONE™ C-17 povidone, PLASDONE™ C-30 povidone, and polyethylene glycol 3350. In yet still further embodiments, the combination of stabilizers may comprise polysorbate 80 and one or more of PLASDONE™ C-12 povidone and povidone K12.
In certain embodiments, the pharmaceutical compositions comprising fulvestrant comprise CMC (carboxymethylcellulose sodium). In some embodiments, the CMC is prepared and heat sterilized before being combined with the fulvestrant during methods of preparation (described more fully elsewhere herein). In further embodiments of the invention, the viscosity of a CMC solution can be modulated by the degree of heating applied, which can allow for the formation of a plurality of fulvestrant pharmaceutical compositions having identical constituents, but with different viscosity values. These different viscosity values can affect the physical stability of the fulvestrant pharmaceutical compositions and the pharmacokinetic characteristics upon administration to subjects. In some embodiments, fulvestrant pharmaceutical compositions comprising CMC may be prepared in two or more parts with each part comprising a different amount of CMC. In other embodiments one or more such parts may be a suspension free of any CMC. In further embodiments, the parts can be mixed in an appropriate ratio to obtain a desired pharmaceutical composition.
In certain embodiments of the invention, the pharmaceutical compositions in the form of liquid suspensions comprising fulvestrant and one or more stabilizers may exhibit different sedimentation behaviors to form either flocculated or caked suspension upon storage. In some embodiments of the invention, after being stored, pharmaceutical compositions in the form of liquid suspensions comprising fulvestrant can be redispersed back into a homogeneous suspension with an acceptable particle size distribution upon redispersion. Exemplary liquid suspension formulations described herein were prepared and tested for sedimentation and redispersion. The tested formulations exhibited different sedimentation behaviors, but all were redispersible back to an acceptable, homogeneous suspension after a 3-month storage period at room temperature.
In certain embodiments of the invention, the pharmaceutical compositions comprising fulvestrant have a pH of from about 3-10, for example, about 3, 4, 5, 6, 7, 8, 9, or about 10. In further embodiments of the invention, the pharmaceutical composition has a pH of from about 5-8. In further embodiments of the invention, the pharmaceutical composition has a pH of from about 6-8. In further embodiments of the invention, the pharmaceutical composition has a pH of from about 3-7. In certain embodiments of the invention, the pharmaceutical composition has a pH of about 6.0 to 8.0. In particular embodiments of the invention, the pharmaceutical composition has a pH of about 6.0 to 7.0, 6.5 to 7.0, 6.5 to 7.5, 6.7 to 7.2, 7.0 to 7.2, 7.0 to 7.5, or 7.0 to 8.0. In further embodiments of the invention, the pharmaceutical composition has a pH of about 7.0.
In particular embodiments of the invention, the pharmaceutical composition further comprises one or more buffering agents, i.e., an agent that when added to a pharmaceutical composition, results in a pharmaceutical composition that resists pH changes or that results in a change in pH, such as, but not limited to, NaH₂PO₄.H₂O, NaH₂PO₄.2H₂O, anhydrous NaH₂PO₄, sodium citrate, citric acid, Tris, sodium hydroxide, HCl, or mixtures thereof. In certain embodiments of the invention, the pharmaceutical composition comprises about 1 mM to 20 mM, of one or more buffering agents, and all ranges and subranges therebetween. In particular embodiments of the invention, the pharmaceutical composition comprises about 1 to 2 mM, 1 to 3 mM, 1 to 5 mM, 2 to 8 mM, 5 to 6 mM, 5 to 10 mM, 8 to 12 mM, 10 to 15 mM, or 15 to 20 mM of one or more buffering agents, and all ranges and subranges there between. In further embodiments of the invention, the pharmaceutical composition comprises about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, or 20 mM of one or more buffering agents.
In certain embodiments of the invention, the pharmaceutical composition has an osmolarity from about 280 mOsm/L to about 310 mOsm/L, for example, about 280, 285, 290, 300, 305, or about 310 mOsm/L. In further embodiments of the invention, the pharmaceutical composition has an osmolarity from about 290 mOsm/L to about 300 mOsm/L. In yet further embodiments of the invention, the pharmaceutical composition has an osmolarity of about 290 mOsm/L. In some embodiments, the osmolarity may be selected through the use of appropriate amounts of the one or more stabilizers, e.g., stabilizers that also act as tonicifiers, such as, but not limited to, the non-electrolyte stabilizers and electrolyte stabilizers described herein. In some embodiments, the osmolarity may be selected through the use of appropriate amounts of one or more buffering agents that act as tonicifiers in a pharmaceutical composition, such as, but not limited to, the buffering agents described herein.
In certain embodiments of the invention, the pharmaceutical composition has an absolute viscosity measured at 25° C. from about 1.0 cP to about 1000 cP, and all ranges and subranges therebetween. In particular embodiments of the invention, the pharmaceutical composition has an absolute viscosity measured at 25° C. from about 750 cP to about 1000 cP, about 500 to about 750 cP, about 250 cP to about 500 cP, about 100 cP to about 250 cP, about 50 cP to about 100 cP, about 25 cP to about 50 cP, about 10 cP to about 25 cP, about 1 cP to about 10 cP, about 1 cP to about 5 cP, about 1.0 cP to about 4.0 cP, about 1.0 cP to about 3.0 cP, about 1.0 cP to about 2.5 cP, about 1.0 cP to about 2.0 cP, about 1.5 cP to about 2.0 cP. In further embodiments of the invention, the pharmaceutical composition has an absolute viscosity measured at 25° C. of about 1.0 cP, 1.1 cP, 1.2 cP, 1.3 cP, 1.4 cP, 1.5 cP, 1.6 cP, 1.7 cP, 1.8 cP, 1.9 cP, 2.0 cP, 2.1 cP, 2.2 cP, 2.3 cP, 2.4 cP, 2.5 cP, 2.6 cP, 2.7 cP, 2.8 cP, 2.9 cP, 3.0 cP, 3.5 cP, 4.0 cP, 4.5 cP, 5.0 cP, 10 cP, 15 cP, or 20 cP.
In yet further embodiments of the invention, pharmaceutical compositions having a fulvestrant concentration of 50 mg/mL or 100 mg/mL have an absolute viscosity measured at 25° C. that is from about 2-fold to about 500-fold lower than FASLODEX™, and all ranges and subranges therebetween. In further embodiments of the invention, fulvestrant pharmaceutical compositions having a fulvestrant concentration of 50 mg/mL or 100 mg/mL have an absolute viscosity measured at 25° C. that is 500-fold lower, about 400-fold lower, about 300-fold lower, about 250-fold lower, about 200-fold lower, about 150-fold lower, about 100-fold lower, about 50-fold lower, about 40-fold lower, about 30-fold lower, about 20-fold lower, about 10-fold lower, about 5-fold lower, about 4-fold lower, about 3-fold lower, about 2-fold lower, or about 1.5-fold lower than FASLODEX™. In further embodiments of the invention, for example, fulvestrant pharmaceutical compositions having a fulvestrant concentration of 50 mg/mL or 100 mg/mL, have an absolute viscosity measured at 25° C. that is substantially equivalent to FASLODEX™. Table 21 provides density measurements of some exemplary fulvestrant pharmaceutical compositions of the present disclosure. Table 22 provides viscosity measurements of some exemplary fulvestrant pharmaceutical compositions of the present disclosure.
In certain embodiments of the invention, the pharmaceutical composition comprises one or more additional pharmaceutically acceptable excipients. As used herein, a pharmaceutically acceptable excipient is generally chemically and/or physically compatible with other ingredients in a pharmaceutical composition or pharmaceutical composition, and/or is generally physiologically compatible with the recipient thereof. In some embodiments, the one or more additional pharmaceutically acceptable excipients are selected from the group consisting of preservatives, antioxidants, or mixtures thereof. In yet further embodiments of the invention, the additional pharmaceutically acceptable excipient is a preservative such as, but not limited to, phenol, cresol, p-hydroxybenzoic ester, chlorobutanol, or mixtures thereof. In yet further embodiments of the invention, the additional pharmaceutically acceptable excipient is an antioxidant such as, but not limited to, ascorbic acid, sodium pyrosulfite, palmitic acid, butylated hydroxyanisole, butylated hydroxytoluene, tocopherols, or mixtures thereof.
In certain embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5.8 mg/mL of one or more stabilizers, and water for injection (WFI) q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 0.8 mg/mL of one or more polymers, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 0.8 mg/mL of povidone K12 (PVP 12K), and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5.8 mg/mL of one or more stabilizers, about 9 mg/mL of one or more electrolytes, about 10 mM of one or more buffering agents, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 0.8 mg/mL of one or more polymers, about 9 mg/mL of sodium chloride, about 10 mM of one or more of NaH₂PO₄.H₂O, NaH₂PO₄.2H₂O, and anhydrous NaH₂PO₄(preferably a mixture of about 0.61 mg/mL NaH₂PO₄.2H₂O and about 0.85 mg/mL of anhydrous NaH₂PO₄), and WFI q.s. to volume
In certain embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 55 mg/mL of one or more stabilizers, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 50 mg/mL of one or more non-electrolytes, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 50 mg/mL of dextrose, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of one or more stabilizers, about 9 mg/mL of one or more electrolytes, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 9 mg/mL of sodium chloride, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 50 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 9 mg/mL of sodium chloride, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 55 mg/mL of one or more stabilizers, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 50 mg/mL of one or more non-electrolytes, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 56.6 mg/mL of one or more stabilizers, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 1.6 mg/mL of one or more polymers, about 50 mg/mL of one or more non-electrolytes, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.6 mg/mL of PLASDONE™ C-12 povidone, povidone K12, or a mixture thereof, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 57.4 mg/mL of one or more stabilizers, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of one or more surfactants, about 2.4 mg/mL of one or more polymers, about 50 mg/mL of one or more non-electrolytes, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 2.4 mg/mL of PLASDONE™ C-12 povidone, povidone K12, or a mixture thereof, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, between about 1 mg/mL and 2.4 mg/mL of PVP, sorbitan palmitate, poloxamer 188, poloxamer 124, poloxamer 427, polyethoxylated castor oil, PVA, or a mixture thereof, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 2.4 mg/mL of PVA, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In yet further embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.0 mg/mL of polyethoxylated castor oil, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 2.0 mg/mL of poloxamer 188, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.5 mg/mL of poloxamer 188, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.5 mg/mL of sorbitan palmitate, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.5 mg/mL of poloxamer 124, about 50 mg/mL of mannitol, and WFI q.s. to volume.
In certain embodiments of the invention, the pharmaceutical composition comprises about 100 mg/mL fulvestrant, about 5 mg/mL of polysorbate 80, about 1.5 mg/mL of poloxamer 407, about 50 mg/mL of mannitol, and WFI q.s. to volume.
Aspects of some exemplary embodiments of pharmaceutical compositions comprising fulvestrant are shown in Tables 1-20 and 23-27.
In the Figures and specification, references are made to exemplary formulations. Some exemplary formulations are identified as “F###” where each “#” is a numeral, e.g., F001, F002, and so on. The exemplary formulations sharing an initial identification “F###” share identical concentrations of constituent components (mg/mL), but may vary in their properties due to different methods of preparation, particle size distributions of fulvestrant, or other differences in processing, storage, or handling. Such exemplary formulation sharing an initial identification scheme “F###” are further identified by extra alphanumeric characters. For example, the exemplary formulations F003a, F003b, and F003k2 have the same concentrations of constituent components but may differ in, e.g., the underlying methods of preparation and resulting particle size distributions. In some instances in the Figures, formulations are identified by only ending non-zero numerals # or ## and subsequent alphanumeric characters; for example, formulation F003a may be referred to as “Variant 3a”, formulation F005a2 may be referred to as “Variant 5a2”, and the like. Some exemplary formulations are identified and referred to as “Lot”s, with references to the same Lot number referring to exemplary formulations having the same concentrations of constituent components, but may vary in their properties due to different methods of preparation, particle size distributions of fulvestrant, or other differences in processing, storage, or handling.

TABLE 1

Target Formulation (mg/mL)	Formulation B	Formulation E	Formulation I

Fulvestrant	50	50	50
Polysorbate 80	5	5	5
PVP 12K	0.8	0.8	0.8
NaCl			9
WFI	q.s. to volume	q.s. to volume	q.s. to volume
Assay (% LC)	100.0	93.5	94.0
Total Impurities (% a/a)	0.2	0.3	0.2

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0 (μm)	LD Dv(10)	7.4	6.0	2.4	1.9	NA	NA
	LD Dv(50)	35.0	32.0	5.2	3.9
	LD Dv(90)	143	129	11.1	8.7

(μm)

day 5

day 1

day 4

LD Dv(10)	7.6	2.3	2.3	1.2	2.5	0.04
LD Dv(50)	31.9	5.7	5.0	1.8	5.6	0.8
LD Dv(90)	107	19.2	10.9	2.5	12.3	3.2

(μm)

day 14

Day 10

Day 11

LD Dv(10)	5.9	5.1	1.9	1.8	2.3	0.06
LD Dv(50)	27.5	24.4	4.0	3.8	4.8	1.3
LD Dv(90)	86.3	76.0	9.1	9.4	9.4	3.7

(μm)

Day 77

Day 71

Day 12

LD Dv(10)	5.4	4.8	2.1	1.8	2.4	0.2
LD Dv(50)	26.3	21.0	4.7	3.8	5.1	3.0
LD Dv(90)	88.7	62.2	12.5	9.7	11.4	6.3

pH	7.4 (day 11)	6.9 (day 7)	7.1 (day 4)
pH	N/A	N/A	6.9 (day 11)

TABLE 2

Target Formulation (mg/mL)	Formulation J	Formulation K	Formulation L

Fulvestrant	50	50	100
Polysorbate 80	5	5	5
PVP 12K	0.8	0.8	0.8
NaCl	9	9	9
Phosphate buffer*	10 mM	10 mM	10 mM
WFI	q.s. to volume	q.s. to volume	q.s. to volume
Assay (% LC)	80.9	82.4	94.1
Total Impurities (% a/a)	0.2	0.2	0.2

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0 (μm)	LD Dv(10)	2.2	1.8	0.04	0.04	2.4	1.9
	LD Dv(50)	5.4	4.2	1.9	1.1	5.0	4.1
	LD Dv(90)	11.8	10.3	4.7	3.6	10.4	8.4

(μm)

day 5

day 4

Day 0

LD Dv(10)	2.0	1.7	0.03	0.04	2.4	1.9
LD Dv(50)	4.7	3.8	1.4	1.1	5.0	4.1
LD Dv(90)	9.7	8.2	3.6	3.0	10.4	8.4

(μm)

Day 12

Day 11

LD Dv(10)	2.1	1.8	1.8	0.05	2.0	2.0
LD Dv(50)	5.2	4.2	3.6	2.1	4.0	4.0
LD Dv(90)	14.8	14.5	6.9	4.9	7.6	7.8

(μm)

Day 13

Day 12

Day 11

LD Dv(10)	2.3	1.9	2.2	0.06	2.5	2.2
LD Dv(50)	5.9	4.6	4.5	2.3	5.3	5.0
LD Dv(90)	16.5	16.0	11.1	5.6	10.8	10.6

pH	7.0 (day 4)	7.0 (day 1)
pH	7.0 (day 11)	7.1 (day 12)

	TABLE 3

	Target Formulation (mg/mL)

	Formulation L3F	Formulation L6

Fulvestrant

	50	100
Polysorbate 80	5	5
PVP 12K	0.8	0.8
NaCl	9	9
Phosphate buffer*	10 mM	10 mM
WFI	q.s. to volume	q.s. to volume
Assay (mg/mL)	83.8	113.9
Total Impurities (% a/a)	0.7	0.2

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

Day 0 (μm)	LD Dv(10)	2.1	2.0	2.2	2.1
	LD Dv(50)	5.9	5.5	6.9	6.7
	LD Dv(90)	14.7	14.2	17.6	17.7

(μm)

Day 14

Day 1

LD Dv(10)	2.1	2.0	2.1	2.1
LD Dv(50)	5.7	5.5	6.8	6.6
LD Dv(90)	14.4	14.0	18.0	17.7

pH			7.1 (day 1)

TABLE 4

Target Formulation	F003a	F003b	F004a

Fulvestrant

	100	100	100
Polysorbate 80	5	5	5
Dextrose	50	50
NaCl			9
WFI	q.s. to volume	q.s. to volume	q.s. to volume
Manufacturing Process	API size reduction by	API size reduction by	API size reduction by
	HSM only	HSM followed by HPH	HSM followed by HPH
Assay (mg/mL)	95.0	96.3	99.8
Total Impurities (%	0.42	0.28	0.27

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0	LD	1.9	1.9	2.2	1.6	1.9	1.5
(μm)	LD	5.3	5.2	6.0	3.9	5.8	3.5
	LD	13.0	12.7	12.1	7.6	12.2	7.7

(μm)

Day 13

LD Dv(10)	1.9	1.9	2.1	1.7	1.7	1.5
LD Dv(50)	5.3	5.2	5.5	4.2	4.0	3.3
LD Dv(90)	13.4	13.0	11.3	8.2	8.7	6.8

(μm)

Day 13

LD Dv(10)	1.8	1.7	2.1	1.7	1.7	1.6
LD Dv(50)	5.1	4.9	5.9	4.3	4.0	3.4
LD Dv(90)	13.0	13.2	12.1	8.4	8.6	6.3

pH	7.3 (Day 0)	7.5 (Day 0)
pH	7.1 (Day 13)	7.1 (Day 13)

TABLE 5

Target Formulation (mg/mL)	F003e	F003k2	F003k3

Fulvestrant	100	100	100
Polysorbate 80	5	5	5
Dextrose	50	50	50
WFI	q.s. to voltune	q.s. to volume	q.s. to volume
Manufacturing Process	Micronized API	Micronized API	Micronized API dispersed
	dispersed by vortex/	dispersed by HSM	by HSM followed by HPH
	sonics or HSM		for size reduction
Assay (mg/mL)	100.5	97.8	99.6
Total Impurities (% a/a)	0.34	0.43	0.42

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0 (μm)	LD Dv(10)	1.5	1.5	1.5	1.4	2.2	1.4
	LD Dv(50)	3.9	3.8	2.9	2.6	7.0	3.4
	LD Dv(90)	13.0	12.8	6.7	6.3	13.3	7.4

(μm)

Day 1

Day 5

Day 1

LD Dv(10)	1.5	1.5	NA	NA	NA	NA
LD Dv(50)	4.0	4.0
LD Dv(90)	12.4	12.5

(μm)

Day 13

Day 12

Day 8

LD Dv(10)	1.5	1.5	1.6	1.5	1.5	1.3
LD Dv(50)	4.1	4.1	3.2	2.9	2.9	2.6
LD Dv(90)	14.0	14.3	7.0	6.5	5.7	5.2

(μm)

Day 22

Day 12

Day 8

LD Dv(10)	1.5	1.5	1.5	1.4	1.4	1.1
LD Dv(50)	3.9	3.9	3.0	2.8	2.8	1.9
LD Dv(90)	12.6	12.8	6.7	6.1	6.2	3.5

pH	7.2 (day 1)	7.2 (day 0)
pH	4.5 (Day 12)	6.8 (Day 8)

TABLE 6

Target Formulation (mg/mL)	F003i	F005a2	F005b1

Fulvestrant

	100	100	100
Polysorbate 80	5	5	5
Dextrose	50
Mannitol		50	50
WFI	q.s. to volume	q.s. to volume	q.s. to volume
Manufacturing Process	Reconstituted lyophilized	Reconstituted lyophilized	Micronized API
	suspension, micronized	suspension, API sized	dispersed by HSM
	API dispersed by HSM	reduction by HSM
Assay (mg/mL)	99.2	93.2	100.2
Total Impurities (% a/a)	0.46	0.55	0.36

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0 (μm)	LD	4.6	1.7	2.1	2.0	1.5	1.4
	LD	54.2	5.6	6.5	5.6	3.2	2.7
	LD	112	16.2	18.4	13.5	7.9	6.6

(μm)

Day 9

Day 8

Day 13

LD	2.5	1.5	2.1	2.0
LD	39.5	4.3	6.2	5.4
LD Dv(90)	95.1	13.5	16.7	13.2

pH	6.5 (Day 3)	7.4 (Day 0)
pH	7.2 (Day 9)

TABLE 7

Target Formulation (mg/mL)	F005c2	F0017a	F0015a

Fulvestrant

	100	100	100
Polysorbate 80	5	15	25
Mannitol	50	50	50
WFI	q.s. to volume	q.s. to volume	q.s. to volume
Manufacturing Process	Micronized API dispersed	Micronized API	Micronized API
	by HSM followed by HPH	dispersed by HSM	dispersed by HSM
	for size reduction
Assay (mg/mL)	100.2		103.8
Total Impurities (% a/a)			0.39

PSD (via laser diffraction)

as is

sonicated

as is

sonicated

as is

sonicated

Day 0 (μm)	LD	1.3	1.1	1.5	1.4	1.5	1.4
	LD	2.3	1.9	3.0	2.6	2.9	2.6
	LD	4.9	3.7	7.2	6.4	6.9	6.4

pH		7.1 (day 0)	7.1 (day 0)

	TABLE 8

	Formulation/Variants

mg/mL	I	J	K	L	M	N	O, O2

Fulvestrant

	50	50	50	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5
Povidone K12	0.8	0.8	0.8	0.8	0.8	0.8	0.8
Sodium Chloride	9	9	9	9	9	4.5
Dextrose						25	50
NaH₂PO₄•2H₂O		0.61	0.61	0.61
Na₂HPO₄		0.85	0.85	0.85
WFI	QS	QS	QS	QS	QS	QS	QS

	TABLE 9

	Formulation

F001

F002

F003

F004

F005

F015

F016

F017

variants (mg/mL)

		F003a, b, e,		F005a, a2, b1,
		f, g, h, i, j,		c, c2, c3, d, g,	F015a,	F017a,
F001e	B, E	k2, k3, l	F004a	d1, g4, g5, h3	a1, a3	a1, a3

Fulvestrant

	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	25	5	15
Povidone K12		0.8					0.8
Sodium Chloride				9
Dextrose			50
Mannitol					50	50	50	50
WFI	QS	QS	QS	QS	QS	QS	QS	QS

	TABLE 10

	Formulation

mg/mL	F003k+	F006	F007	F008	F009	F010	F011	F012	F013	F014

Fulvestrant

	200	100	100	100	100	100	100	100	100	100
Polysorbate 80	5	25	5	5	5	5	5	5	5	5
Dextrose	50	50	50	50	50	50	50	50	50	50
Polysorbate 20			15
Poloxamer 188				2
PVP K12					20
PVP K17						5
PEG 3350							60
CMC7LF PH								30
CMC7MF PH									20
CMC7HF PH										5
WFI	QS	QS	QS	QS	QS	QS	QS	QS	QS	QS

TABLE 11

(mg/mL)	F018	F019	F020	F021	F022	F023	F024	F025	F026	F027	F028

Fulvestrant

	100	100	100	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5	5	5	5	5
Polysorbate 20					5	15		5	15	5
Poloxamer 188							2	2	2	2
Mannitol	50	50	50	50	50	50	50	50	50	50	50
Dextrose
NaCl
Glycerol
PVP K12
	5	10	20							10
PVP K17				5
PEG 3350											60
CMC7LF PH
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs	qs	qs	qs	qs

TABLE 12

(mg/mL)	F029	F030	F031	F032	F033	F034	F035	F036

Fulvestrant

	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5	5
Polysorbate 20								5
Poloxamer 188								2
Mannitol	50	25	12.5	37.5	50	50	50	50
Dextrose		25	37.5	12.5
NaCl
Glycerol
	30
PVP K12								5
PVP K17
PEG 3350
CMC7LF PH					3	2	1	1
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs	qs

TABLE 13

(mg/mL)	F037	F038	F039	F040	F041	F042	F043	F044	F045	F046	F047

Fulvestrant

	100	100	100	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5	5	5	5	5
Polysorbate 20				5				5
Poloxamer 188				2				2
Mannitol	25	25	25	25	12.5	12.5	12.5	12.5	37.5	37.5	37.5
Dextrose	25	25	25	25	37.5	37.5	37.5	37.5	12.5	12.5	12.5
NaCl
Glycerol
PVP K12
				5				5
PVP K17
PEG 3350
CMC7LF PH	3	2	1	1	3	2	1	1	3	2	1
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs	qs	qs	qs	qs

TABLE 14

(mg/mL)	F048	F049	F050	F051	F052	F053	F054	F055

Fulvestrant*	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5	5
Polysorbate 20	5							5
Poloxamer 188	2							2
Mannitol	37.5	25	25	25	25	25	25	25
Dextrose	12.5			25	25	25	25	25
NaCl		4.9	9	4.9	4.9	4.9	4.9	4.9
Glycerol
PVP K12
	5							5
PVP K17
PEG 3350
CMC7LF PH	1				3	2	1	1
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs	qs

TABLE 15

(mg/mL)	F056	F057	F058	F059	F060	F061	F062

Fulvestrant

	100	100	100	100	100	100	100
Polysorbate 80
Polysorbate 20	5	10	15	5	5	10	10
Poloxamer 188
Mannitol	50	50	50	50	50	50	50
Dextrose
NaCl
Glycerol
PVP K12
PVP K17
PEG 3350				10	30	10	30
PEG 4000
CMC7LF PH
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs

TABLE 16

(mg/mL)	F063	F064	F065	F066	F067	F068	F069	F070

Fulvestrant

	100	100	100	100	100	100	100	100
Polysorbate 80
Polysorbate 20	15	15	5	5	10	10	15	15
Poloxamer 188
Mannitol	50	50	50	50	50	50	50	50
Dextrose
NaCl
Glycerol
PVP K12
PVP K17
PEG 3350	10	30
PEG 4000			3	7.5	3	7.5	3	7.5
CMC7LF PH
CMC7MF PH
CMC7HF PH
WFI	qs	qs	qs	qs	qs	qs	qs	qs

	TABLE 17

	Formulations/Variants

Component	Q	R	S	T	U	V	W	X

Fulvestrant (mg/mL)	100	100	100	100	100	100	100	100
Polysorbate 80	25	5	—	—	5	5	—	5
(mg/mL)
Polysorbate 20	—	—	5	—	—	—	—	—
(mg/mL)
Poloxamer 188	—	—	—	5	—	—	—	—
(mg/mL)
Lecithin (mg/mL)	—	—	—	—	—	—	5	—
PVP K12 (mg/mL)	0.8	5	0.8	0.8	—	—	0.8	0.8
PVP K17 (mg/mL)	—	—	—	—	0.8	—	—	—
PEG 3350 (mg/mL)	—	—	—	—	—	50	—	—

Dextrose (mg/mL)	50	—	—
Sodium Chloride	—	9	13
(mg/mL)

NaOH	QS to	QS to	QS to	QS to	QS to	QS to	QS to pH	QS to
	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	7.0	pH 7.0
HCl	QS to	QS to	QS to	QS to	QS to	QS to	QS to pH	QS to
	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	7.0	pH 7.0
WFI	QS to	QS to	QS to	QS to	QS to	QS to	QS	QS to
	volume	volume	volume	volume	volume	volume	to volume	volume

	TABLE 18

	Formulations/Variants

Component

	1	2	3	4	5	6	7	8

Fulvestrant	100	100	100	100	100	100	100	100
(mg/mL)
Polysorbate 80	—	5	—	—	5	5	—	5
(mg/mL)
Polysorbate 20	—	—	5	—	—	—	—	—
(mg/mL)
Poloxamer 188	—	—	—	5	—	—	—	—
(mg/mL)
Human Serum	5
Albumin (mg/mL)
Lecithin (mg/mL)	—	—	—	—	—	—	5	—
PVP K12	0.8	5	0.8	0.8	—	—	0.8	0.8
(mg/mL)
PVP K17	—	—	—	—	0.8	—	—	—
(mg/mL)
PEG 3350	—	—	—	—	—	50	—	—
(mg/mL)

Dextrose	50	50	—
(mg/mL)
Mannitol (mg/mL)	—	—	50

Citric buffer	QS to	QS to	QS to	QS to	QS to	QS to	QS to pH	QS to
	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	pH 7.0	7.0	pH 7.0
WFI	QS to	QS to	QS to	QS to	QS to	QS to	QS	QS to
	volume	volume	volume	volume	volume	volume	to volume	volume

	TABLE 19

	Formulation (mg/mL)

	F005g4		Lot	Lot	Lot	Lot	Lot	Lot	Lot
	F005g5	Lot
15	26	27	28	42	43	X1	X2

Fulvestrant

	100	100	100	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5	5	5	5
PVPC12	—		—	1.6	1.6	—	—	—	—
Span 40	—		—	—	—	1.5	—	—	—
Pluronic F-68							1.5
Pluronic L44								1.5
Pluronic F127									1.5
Mannitol (before	50	50	—	50	—	50	50	50	50
homogenization)
Mannitol (after	—		50	—	50
homogenization)

	TABLE 20

	Formulation (mg/mL)

Lot	Lot	Lot	Lot
45	46	47	48

Fulvestrant	100	100	100	100
Polysorbate 80	5	5	5	5
PVPC12	2.4	—
Span 40	—	—
Pluronic F-68		2
Cremophor EL			1
PVA				2.4
Mannitol	50	50	50	50
(before
homogenization)

	TABLE 21

	Sample	Density
	Name	(g/ml)

	F003h	1.032
	F003f	1.032
	F003e	1.032
	F003k2	1.030

TABLE 22

Sample		Viscosity
Name	Formulation	(cps)

Placebo	5 mg/mL PS80 + 50 mg/mL	1.1
	Dextrose
F003h	[Described elsewhere]	1.8
F003f	[Described elsewhere]	1.9
F003k	[Described elsewhere]	2.0
F003e	[Described elsewhere]	1.5

B. Fulvestrant Particles

Particular embodiments of the disclosure comprise solid fulvestrant particles, for example a fulvestrant suspension comprising solid fulvestrant particles. In certain embodiments of the invention, at least about 90% of the total fulvestrant in the formulation is present as solid particles. In further embodiments of the invention, at least about 80% of the total fulvestrant in the formulation is present as solid particles.
In certain embodiments of the invention, solid fulvestrant particles are particles consisting of crystalline and/or amorphous fulvestrant. In other embodiments of the invention, fulvestrant particles comprise crystalline and/or amorphous fulvestrant as well as other excipients. In still other embodiments, fulvestrant particles comprise crystalline and/or amorphous fulvestrant coated or surface modified by a surface modifier adsorbed on the surface of the particle. The surface modifier can be a stabilizer such as, but not limited to surfactants, polymers, electrolytes, and non-electrolytes, and mixtures thereof.
Other embodiments of the present invention may further comprise fulvestrant in forms other than a solid particle, such as, but not limited to, solubilized fulvestrant as a free molecule or associated with a suspension such as micelles, microemulsions, emulsion, liposome, and combinations thereof, or complexed with other formulation constituents in a vehicle. In further embodiments of the invention, such other forms of fulvestrant are in equilibrium with the fulvestrant solid particles.
In particular embodiments of the invention, the fulvestrant particles comprise about 90-99.9% by weight of fulvestrant and 0.1-10% by weight of a surface modifier adsorbed on the surface of said particle. In particular embodiments of the invention, the surface modifier is a stabilizer such as, but not limited to surfactants, polymers, electrolytes, and non-electrolytes, and mixtures thereof. In certain embodiments of the invention, fulvestrant particles comprise at least about 90% fulvestrant. In other embodiments of the invention fulvestrant particles comprise at least about 92%, 95%, 97%, 98%, 99%, 99.5%, or 99.9% fulvestrant.
In further embodiments of the invention, one or more solvents, such as water, present in the pharmaceutical composition can be removed partially or completely by appropriate techniques known to the art, such as lyophilization or spray drying, to form a dried pharmaceutical composition for reconstitution. In certain embodiments of the invention, the dried pharmaceutical composition can comprise up to about 1%, about 2%, about 5%, or about 10% of the one or more solvents. The dried pharmaceutical composition can be reconstituted with appropriate diluent known to the art, such as, but not limited to water for injection (WFI), normal saline (NS), and 5% dextrose in water (D5W) prior to administration. In further embodiments of the invention, the diluent can further comprise an organic solvent or one or more of the excipients described herein. Dried pharmaceutical compositions formed by lyophilization may be in the form of a lyophilized cake.

Fulvestrant Particle Sizes

In certain embodiments of the invention, the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In yet further embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 micron. In other embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In still other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns.
In certain embodiments of the invention, the fulvestrant particles have a laser diffraction diameter greater than or equal to about 0.5 microns. In other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 0.5 microns. In other embodiments of the invention, the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 microns. In still other embodiments of the invention, the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1.5 microns. In other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1.5 microns. In yet other embodiments of the invention, the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns.
In further embodiments of the invention, about 98% of fulvestrant particles have a laser diffraction diameter greater than or equal to about 0.5 microns. In other embodiments of the invention, about 98% of fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In still other embodiments of the invention, about 98% of fulvestrant particles have a laser diffraction diameter greater than or equal to about 1.5 microns. In yet other embodiments of the invention, about 98% of fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) between about 4 microns and about 120 microns, between about 4 microns and about 100 microns, between about 4 microns and about 75 microns, between about 4 microns and about 60 microns, between about 4 microns and about 50 microns, between about 4 microns and about 40 microns, between about 4 microns and about 30 microns, between about 4 microns and about 20 microns, between about 4 microns and about 15 microns, between about 4 microns and about 10 microns, between about 20 microns and about 60 microns, between about 20 microns and about 45 microns, between about 20 microns and about 30 microns, between about 30 microns and about 50 microns, or between about 4 microns and about 9 microns. In other embodiments of the invention, the fulvestrant particles have a LD Dv(90) equal to about 4 microns, about 5 microns, about 6 microns, about 7 microns, about 8 microns, about 9 microns, about 10 microns, about 11 microns, about 12 microns, about 13 microns, about 14 microns, about 15 microns, about 16 microns, about 17 microns, about 18 microns, about 19 microns, about 20 microns, about 25 microns, about 30 microns, about 35 microns, about 40 microns, about 45 microns, about 50 microns, about 55 microns, about 60 microns, about 65 microns, about 70 microns, about 75 microns, about 80 microns, about 85 microns, about 90 microns, about 95 microns, about 100 microns, about 105 microns, about 110 microns, about 115 microns, or about 120 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 120 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 100 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 80 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 60 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 50 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 40 microns. In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 30 microns. In further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 25 microns. In further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 18 microns. In further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 16 microns. In further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 14 microns. In still further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 11 microns. In yet further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 9 microns. In yet further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 7 microns. In yet further embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 5 microns. In particular embodiments of the invention, particles have an LD Dv(90) between about 9-14 microns. In other embodiments of the invention, the particles have an LD Dv(90) between about 12-14 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 9-11 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 7-9 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 6-8 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 6-7 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 3-6 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(50) between about 2 microns and about 35 microns, between about 2 microns and about 25 microns, between about 2 microns and about 20 microns, between about 2 microns and about 15 microns, between about 2 microns and about 10 microns, between about 2 microns and about 8 microns, between about 2 microns and about 7 microns, between about 2 microns and about 6 microns, between about 2 microns and about 5 microns, between about 2 microns and about 4 microns, between about 5 microns and about 10 microns, between about 5 microns and about 15 microns, between about 7 microns and about 10 microns, between about 8 microns and about 10 microns, or between about 9 microns and about 16 microns. In other embodiments of the invention, the fulvestrant particles have a LD Dv(50) equal to about 2 microns, 3 microns, 4 microns, about 5 microns, about 6 microns, about 7 microns, about 8 microns, about 9 microns, about 10 microns, about 11 microns, about 12 microns, about 13 microns, about 14 microns, about 15 microns, about 16 microns, about 17 microns, about 18 microns, about 19 microns, about 20 microns, about 25 microns, about 30 microns, or about 35 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(50) less than or equal to about 9 microns. In other embodiments of the invention, the particles have an LD Dv(50) less than or equal to about 7 microns. In other embodiments of the invention, the particles have an LD Dv(50) less than or equal to about 6 microns. In yet other embodiments of the invention, the particles have an LD Dv(50) less than or equal to about 5 microns. In particular embodiments of the invention, the particles have an LD Dv(50) less than or equal to about 4 microns. In further embodiments of the invention, the particles have an LD Dv(50) less than or equal to about 3 microns. In further embodiments of the invention, the particles have an LD Dv(50) between about 4-6 microns. In further embodiments of the invention, the particles have an LD Dv(50) between about 3-5 microns. In yet further embodiments of the invention, the particles have an LD Dv(50) between about 3-4 microns. In yet further embodiments of the invention, the particles have an LD Dv(50) between about 2-3 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(10) no more than about 3 microns, about 2 microns, or about 1 microns. In further embodiments of the invention, the particles have an LD Dv(10) between about 1 micron and about 3 microns. In still further embodiments of the invention, the particles have an LD Dv(10) greater than or equal to about 2 microns. In yet further embodiments of the invention, the particles have an LD Dv(10) between about 1.5 microns to about 2.5 microns. In yet further embodiments of the invention, the particles have an LD Dv(10) between about 1 micron to about 2 microns. In yet further embodiments of the invention, the particles have an LD Dv(10) between about 1.0 micron to about 1.5 microns. In even further embodiments of the invention, the particles have an LD Dv(10) of about 2 microns. In even further embodiments of the invention, the particles have an LD Dv(10) of about 1.5 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) less than or equal to about 25 microns and an LD Dv(50) less than or equal to about 9 microns. In particular embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 16 microns and an LD Dv(50) less than or equal to about 6 microns. In other embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 11 microns and an LD Dv(50) less than or equal to about 5 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 9 microns and an LD Dv(50) less than or equal to about 4 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) less than or equal to about 8 microns and an LD Dv(50) less than or equal to about 4 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) between about 9-14 microns and an LD Dv(50) between about 4-6 microns. In still other embodiments of the invention, the particles have an LD Dv(90) between about 9-11 microns and an LD Dv(50) between about 4-6 microns. In particular embodiments of the invention, the particles have an LD Dv(90) between about 12-14 microns and an LD Dv(50) between about 4-6 microns. In further embodiments of the invention, the particles have an LD Dv(90) between about 6-8 microns and an LD Dv(50) between about 2-4 microns. In further embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In yet further embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 micron. In other embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In still other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) between about 9-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns. In other embodiments of the invention, the particles have an LD Dv(90) between about 9-11 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 12-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 6-9 microns, an LD Dv(50) between about 2-4 microns, and an LD Dv(10) between about 1-2 microns. In further embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In yet further embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 micron. In other embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In still other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns.
In certain embodiments of the invention, the fulvestrant particles have an LD Dv(90) between about 9-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns, and the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In other embodiments of the invention, the particles have an LD Dv(90) between about 9-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns, and at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 micron. In yet other embodiments of the invention, the fulvestrant particles have an LD Dv(90) between about 30 microns and about 110 microns, an LD Dv(50) between about 5 microns and about 30 microns, and an LD Dv(10) between about 1.5 microns and about 3 microns. In other embodiments of the invention, the particles have an LD Dv(90) between about 9-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns, and the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In still other embodiments of the invention, the particles have an LD Dv(90) between about 9-14 microns, an LD Dv(50) between about 4-6 microns, and an LD Dv(10) between about 2-3 microns, and at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 6-9 microns, an LD Dv(50) between about 2-4 microns, an LD Dv(10) between about 1-2 microns, and the fulvestrant particles have a laser diffraction diameter greater than or equal to about 0.5 microns. In yet other embodiments of the invention, the particles have an LD Dv(90) between about 6-9 microns, an LD Dv(50) between about 2-4 microns, an LD Dv(10) between about 1-2 microns, and at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 0.5 microns. In further embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 1 micron. In yet further embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 1 micron. In other embodiments of the invention the fulvestrant particles have a laser diffraction diameter greater than or equal to about 2 microns. In still other embodiments of the invention, at least a portion of the fulvestrant particles have a laser diffraction diameter less than about 2 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dv(10) between about 1 microns and about 25 microns, between about 2 microns and about 25 microns, between about 3 microns and about 7 microns, between about 4 microns and about 15 microns, between about 4 microns and about 10 microns, between about 4 microns and about 8 microns, between about 6 microns and about 8 microns, between about 6 microns and about 7 microns, or between about 1 microns and about 10 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dv(10) equal to about 1 micron, about 2 microns, about 3 microns, about 4 microns, about 5 microns, about 6 microns, about 7 microns, about 8 microns, about 9 microns, about 10 microns, about 11 microns, about 12 microns, about 13 microns, about 14 microns, about 15 microns, about 16 microns, about 17 microns, about 18 microns, about 19 microns, about 20 microns, about 21 microns, about 22 microns, about 23 microns, about 24 microns, or about 25 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dv(50) between about 5 microns and about 60 microns, between about 5 microns and about 50 microns, between about 9 microns and about 20 microns, between about 9 microns and about 15 microns, between about 10 microns and about 50 microns, between about 10 microns and about 40 microns, between about 10 microns and about 30 microns, between about 10 microns and about 20 microns, between about 15 microns and about 30 microns, between about 15 microns and about 25 microns, between about 15 microns and about 20 microns, or between about 10 microns and about 15 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dv(50) equal to about 5 micron, about 6 microns, about 7 microns, about 8 microns, about 9 microns, about 10 microns, about 11 microns, about 12 microns, about 13 microns, about 14 microns, about 15 microns, about 16 microns, about 17 microns, about 18 microns, about 19 microns, about 20 microns, about 21 microns, about 22 microns, about 23 microns, about 24 microns, about 25 microns, about 30 microns, about 35 microns, about 40 microns, about 45 microns, about 50 microns, about 55 microns, or about 60 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dv(90) between about 10 microns and about 200 microns, between about 25 microns and about 150 microns, between about 25 microns and about 125 microns, between about 25 microns and about 100 microns, between about 25 microns and about 75 microns, between about 25 microns and about 50 microns, between about 25 microns and about 40 microns, between about 25 microns and about 35 microns, between about 35 microns and about 90 microns, between about 35 microns and about 75 microns, between about 35 microns and about 50 microns, between about 35 microns and about 45 microns, between about 50 microns and about 100 microns, between about 50 microns and about 75 microns, or between about 20 microns and about 40 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dv(90) equal to about 10 microns, about 15 microns, about 20 microns, about 25 microns, about 30 microns, about 35 microns, about 40 microns, about 45 microns, about 50 microns, about 55 microns, about 60 microns, about 65 microns, about 70 microns, about 75 microns, about 80 microns, about 85 microns, about 90 microns, about 95 microns, about 100 microns, about 105 microns, about 110 microns, about 115 microns, about 120 microns, about 125 microns, about 130 microns, about 135 microns, about 140 microns, about 145 microns, about 150 microns, about 155 microns, about 160 microns, about 165 microns, about 170 microns, about 175 microns, or about 200 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dv(90) between about 35 microns and about 90 microns, a CE Dv(50) between about 10 microns and about 35 microns, and a CE Dv(10) between about 4 microns and about 10 microns. In other embodiments of the invention, the particles have a CE Dv(90) between about 25 microns and about 60 microns, a CE Dv(50) between about 10 microns and about 25 microns, and a CE Dv(10) between about 4 microns and about 8 microns. In other embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 35 microns, a CE Dv(50) between about 10 microns and about 20 microns, and a CE Dv(10) between about 4 microns and about 8 microns. In still other embodiments of the invention, the particles have a CE Dv(90) between about 30 microns and about 100 microns, a CE Dv(50) between about 10 microns and about 50 microns, and a CE Dv(10) between about 4 microns and about 10 microns. In yet other embodiments of the invention, the particles have a CE Dv(90) between about 50 microns and about 100 microns, a CE Dv(50) between about 20 microns and about 50 microns, a CE Dv(10) between about 6 microns and about 8 microns. In yet other embodiments of the invention, the particles have a CE Dv(90) between about 50 microns and about 75 microns, a CE Dv(50) between about 30 microns and about 40 microns, a CE Dv(10) between about 8 microns and about 10 microns. In yet other embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 60 microns, a CE Dv(50) between about 9 microns and about 20 microns, and a CE Dv(10) between about 3 microns and about 7 microns. In still further embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 50 microns, a CE Dv(50) between about 9 microns and about 20 microns, and a CE Dv(10) between about 3 microns and about 7 microns. In other embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 45 microns, a CE Dv(50) between about 9 microns and about 20 microns, and a CE Dv(10) between about 3 microns and about 7 microns. In yet further embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 40 microns, a CE Dv(50) between about 9 microns and about 15 microns, and a CE Dv(10) between about 3 microns and about 7 microns. In further embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 35 microns, a CE Dv(50) between about 9 microns and about 15 microns, and a CE Dv(10) between about 3 microns and about 7 microns. In still other embodiments of the invention, the particles have a CE Dv(90) between about 20 microns and about 45 microns, a CE Dv(50) between about 9 microns and about 15 microns, and a CE Dv(10) between about 3 microns and about 7 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dn(90) between about 4 microns and about 20 microns, between about 6 microns and about 15 microns, between about 6 microns and about 12 microns, between about 8 microns and about 12 microns, between about 8 microns and about 11 microns, between about 4 microns and about 10 microns, between about 4 microns and about 8 microns, between about 4 microns and about 7 microns, or between about 4 microns and about 6 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dn(90) equal to about 4 microns, about 5 microns, about 6 microns, about 7 microns, about 8 microns, about 9 microns, about 10 microns, about 11 microns, about 12 microns, about 13 microns, about 14 microns, about 15 microns, about 16 microns, about 17 microns, about 18 microns, about 19 microns, or about 20 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dn(50) between about 2.0 microns and about 10.0 microns, between about 2.0 microns and about 8.0 microns, between about 2.0 microns and about 6.0 microns, between about 2.0 microns and about 5.0 microns, between about 3.0 microns and about 5.0 microns, between about 3.5 microns and about 4.5 microns, between about 2.0 microns and about 4.0 microns, between about 2.5 microns and about 4.5 microns, or between about 2.5 microns and about 3.5 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dn(50) equal to about 2.0 microns, about 2.5 microns, about 3.0 microns, about 3.5 microns, about 4.0 microns, about 4.5 microns, about 5.0 microns, about 5.5 microns, about 6.0 microns, about 6.5 microns, about 7.0 microns, about 7.5 microns, about 8.0 microns, about 8.5 microns, about 9.0 microns, about 9.5 microns, or about 10.0 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dn(10) between about 0.5 microns and about 2.0 microns, between about 0.5 microns and about 1.5 microns, between about 1.0 microns and about 1.5 microns, between about 0.8 microns and about 1.2 microns, between about 0.9 microns and about 1.1 microns, or between about 0.5 microns and about 1.0 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dn(10) equal to about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1.0, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, or about 2.0 microns.
In certain embodiments of the invention, the fulvestrant particles have a CE Dn(90) between about 4 microns and about 20 microns, a CE Dn(50) between about 2.0 microns and about 10.0 microns, and a CE Dn(10) between about 0.5 microns and about 2.0 microns. In other embodiments of the invention, the fulvestrant particles have a CE Dn(90) between about 6 microns and about 12 microns, a CE Dn(50) between about 2.0 microns and about 6.0 microns, and a CE Dn(10) between about 0.5 microns and about 1.5 microns. In further embodiments of the invention, the fulvestrant particles have a CE Dn(90) between about 8 microns and about 1 microns, a CE Dn(50) between about 3.0 microns and about 5.0 microns, and a CE Dn(10) between about 0.8 microns and about 1.2 microns.

C. Preparation Methods

In certain embodiments of the invention, formulations of the invention can be prepared from commercially available fulvestrant having different particle size distributions, such as, for example, recrystallized, micronized fulvestrant, or a combination thereof. In further embodiments of the invention, the formulations are prepared with sterilized, commercially available fulvestrant. In particular embodiments, commercially available fulvestrant is used in the formulations of the present invention without further processing for size reduction.
In other embodiments of the invention, fulvestrant particles suitable for use in formulations of the invention can be prepared from commercially available fulvestrant by any suitable methods known in the art. Suitable methods include, but are not limited to, size-reduction techniques such as milling, grinding, crushing, compression, attrition, low shear mixing, high shear mixing, high pressure homogenization, lyophilization, precipitation, or combinations thereof.
Desired particle size distributions for fulvestrant particles can be achieved by processing steps at one or more stages of formulation preparation. In some embodiments, the desired particle size distribution can be formed by processing fulvestrant material prior to suspension in media, by techniques described more fully elsewhere herein. In other embodiments, the desired particle size distribution can be formed by processing after suspension in media, by techniques described more fully elsewhere herein, including but not limited to high shear mixing and high pressure homogenization. In still other embodiments, the desired particle size distribution can be formed by a combination of the processing prior to and after suspension in media.
Suitable milling techniques include, but are not limited to, dry milling, wet milling, and cryogenic milling. Suitable milling machines include ball mills, pebble mills, rod mills, roller mills, colloid mills, impact mills, and jet mills. In certain embodiments of the invention, the particles can be reduced in size in the presence of one or more excipients or stabilizers, such as but not limited to a surfactants, polymers, electrolytes, and non-electrolytes, and mixtures thereof. Alternatively, the particles can be contacted with one or more excipients or stabilizers after they are reduced in size.
In certain embodiments of the invention, the formulations can be prepared from an un-milled commercially available fulvestrant by reducing the fulvestrant particle size with high shear mixing. In yet further embodiments of the invention, the formulations can be prepared from an un-milled, commercially available fulvestrant by reducing the fulvestrant particle size with high shear mixing followed by high pressure homogenization.
In certain embodiments of the invention, the formulations can be prepared from commercially available micronized fulvestrant by reducing the micronized fulvestrant particle size with high shear mixing (“HSM”). In yet further embodiments of the invention, the formulations can be prepared from an un-milled commercially available fulvestrant by reducing the fulvestrant particle size with high shear mixing followed by high pressure homogenization (“HPH”).
In some embodiments of the invention, the formulations can be prepared using high pressure homogenization. In further embodiments of the invention, the high pressure homogenization process reduces particle size by subjecting the particle population to one or more of cavitation, shear, and impact within a homogenization chamber under operating pressures from about 5,000 psi to about 45,000 psi, for example, about 5,000, 10,000, 15,000, 20,000, 25,000, 30,000, 35,000, 40,000 or about 45,000 psi. In yet further embodiments of the invention, the high pressure homogenization process is performed at about 40,000 psi. In some embodiments of the invention, the high pressure homogenization process is performed at operating pressures from about 15,000 psi to about 20,000 psi. In further embodiments, the formulations can be prepared by passing the formulation suspension through a homogenization chamber under operating pressures for one or more passes, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, or 50 passes.
In the Figures and specification, references are made to exemplary formulations and processes for preparing exemplary formulations. Some exemplary preparation processes are identified with alphanumeric reference identifiers, such as “Process A1,” “Process A2,” and so on. Some exemplary formulations can share identical concentrations of constituent components (mg/mL), but may vary in their properties due to the different preparation processes, storage, or handling, which can result in different particle size distributions due to more or less size reduction, more or less aggregation or agglomeration, or both during processing, storage, or handling.

Methods of Forming Aqueous Fulvestrant Suspensions

In some embodiments of the invention, methods of forming an aqueous fulvestrant suspension comprise mixing an aqueous medium and at least one stabilizer to form a suspension vehicle, adding an amount of fulvestrant to the suspension vehicle, and dispersing the fulvestrant in the suspension vehicle to form the aqueous fulvestrant suspension. In further embodiments, these methods can further comprise homogenizing the aqueous fulvestrant suspension. In yet further embodiments, the methods with or without the homogenizing step can further comprise concentrating the fulvestrant suspension by phase separating the suspension and removing a portion of the supernatant. In particular further embodiments, after the concentrating step the methods can further comprise adding one or more electrolytes, non-electrolytes, buffering agents, or cross-linked polymers to the homogenized aqueous fulvestrant suspension and mixing the one or more electrolytes, non-electrolytes, buffering agents, or cross-linked polymers into the suspension. In some embodiments of the invention, the methods comprise a dispersing step performed using high shear mixing, a homogenizing step performed using high pressure homogenization, or both a dispersing step performed using high shear mixing and a homogenizing step performed using high pressure homogenization.
In further embodiments of the invention that include one or more stabilizers, the one or more stabilizers may be incorporated into the formulations at one or more stages of the methods of forming the formulations. In some embodiments, at least a portion or all of the stabilizers of a formulation are added to an aqueous medium along with an amount of fulvestrant prior to some or all of any mixing, homogenization, or supernatant-removal steps. In still other embodiments, at least a portion or all of the one or more stabilizers of a formulation are added to the fulvestrant suspensions after some or all of any mixing, homogenization, or supernatant-removal steps have been completed. In further embodiments, at least a portion or all of the surfactant and polymer stabilizers of the formulations are combined with the aqueous medium and fulvestrant prior to some or all of any mixing, homogenization, or supernatant-removal steps and at least a portion or all of the electrolyte, non-electrolyte, buffering agents, and cross-linked polymers of the formulations are added to the suspension after some or all of any mixing, homogenization, or supernatant-removal steps.
The fulvestrant particles described herein can be prepared in a method comprising the steps of dispersing fulvestrant particles in a liquid suspension medium and applying mechanical means in the presence of grinding media to reduce the particle size of fulvestrant to the desired size.
In further embodiments of the invention, a solvent, such as water, present in a formulation can be removed by appropriate techniques known to the art, such as lyophilization or spray drying, to form a dried formulation suitable for later reconstitution. Lyophilization can be used to produce a lyophilized (lyo) cake. The dried formulation can be reconstituted back into a liquid suspension using an appropriate diluent. Different volumes of diluent can be used to produce reconstituted suspensions with different fulvestrant concentrations as needed. The diluent can be aqueous in general but can further comprise an organic solvent and/or any excipient as described elsewhere herein.
In some embodiments of the invention, at least a portion of the formulation components other than fulvestrant can be omitted from the suspension and incorporated as part of the diluent and introduced into the suspension upon reconstitution by the diluent to arrive at the final formulation. In further embodiments, suspensions can be prepared with higher or lower concentrations of constituent components than desired in formulations for administration, formed into dried formulations and placed into vials in appropriate amounts of dried formulation to achieve target dose amounts of fulvestrant per vial for later reconstitution of diluent to form the desired formulation for administration.
Some exemplary methods of preparation of dried pharmaceutical compositions are depicted schematically in FIG. 15.
In some embodiments, the pharmaceutical compositions and dried pharmaceutical compositions can be prepared using aseptic process or terminally sterilized by a compatible sterilization technique, such as, but not limited to, gamma irradiation. When a polymer is used as an excipient in the pharmaceutical composition, said polymer such as microcrystalline cellulose (CMC) or its salts including sodium CMC, can be sterilized by autoclave in a solution then combined with rest of the pharmaceutical composition that is prepared aseptically or terminally sterilized.
Some aspects of exemplary embodiments of methods of preparation of the invention are shown in Tables 4-7 and 23-27 and FIGS. 4-12 and 15 which describe aspects of the preparation methods for exemplary fulvestrant formulations.

	TABLE 23

	Target Formulation (mg/mL)

	B	E	I	J	K	L

Fulvestrant	50	50	50	50	50	100
Polysorbate 80	5	5	5	5	5	5
PVP 12K	0.8	0.8	0.8	0.8	0.8	0.8
NaCl	—	—	9	9	9	9
Phosphate	—	—	—	10 mM	10 mM	10 mM
buffer
WFI	q.s. to	q.s. to	q.s. to	q.s. to	q.s. to volume	q.s. to volume
Starting API	Un-milled	Un-milled API	Un-milled API	Un-milled API	Un-milled API	Un-milled API
(PSD via laser	API	L D Dv90: 780	L D Dv90: 780	L D Dv90: 1890	L D Dv90: 1890	L D Dv90: 1890
diffraction)	L D Dv90:	microns	microns	microns	microns	microns
	780
Manufacturing	API size	API size	API size	API size	API size reduction	API size reduction by
process	reduction	reduction by	reduction by	reduction by	by HSM followed	HSM followed by
	by HSM	HSM followed	HSM followed	HSM followed	by HPH in the	HPH in the presence
		by HPH	by HPH, then	by HPH in the	presence of salts,	of salts
			subsequent salt	presence of	then further HSM
			addition	salts
Process End	L D Dv90:	Total 15 min	Total 42 min	Total 45 min	Total 45 min HSM	Total 25 min HSM at
Point Targets	143	HSM at	HSM at	HSM at	at	~25,000 rpm
(PSD via laser	micron	~20,000 rpm	~20,000 rpm	~25,000 rpm	~25,000 rpm	(L D Dv90:<~80
diffraction)	HSM:	(L D Dv90:<~80	(L D Dv90:<~40	(L D Dv90:<~50	(L D Dv90:<~50	micron)
	Total 15 min	micron)	micron)	micron)	micron)	L D Dv90: 13.8
	at	HPH: 15 passes	L D Dv90: 12.6	L D Dv90: 11.8	L D Dv90: 4.7	microns
	~20,000 rpm	in reverse flow	micron	micron	micron	HPH: 15 passes in
		through z5	HPH: 30 passes	HPH: 3 passes	HPH: 12 passes in	parallel flow through
		nozzle at	in parallel flow	in parallel flow	parallel flow	z8 nozzle at ~30,000 psi
		~30,000 psi	through z5	through z5	through z5 nozzle at
		L D Dv90: 11.1	nozzle at	nozzle at	~40,000 psi to
		micron	~40,000 psi	~40,000 psi	L D Dv90: ~8
					micron then
					additional 5 min
					HSM at ~25,000 rpm

	TABLE 24

	Target Formulation (mg/mL)

	L3F	L6	F003a	F003b	F004a	F003e

Fulvestrant	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	5	5
PVP 12K	0.8	0.8	—	—	—	—
NaCl	9	9	—	—	—	—
Phosphate	10 mM	10 mM	—	—	9	—
buffer
Dextrose	—	—	50	50	—	50
WFI	q.s. to	q.s. to	q.s. to	q.s. to	q.s. to	q.s. to
Starting API	Un-milled	Un-milled	Un-milled API	Un-milled API	Un-milled API	Jet-milled
(PSD via laser	API	API	L D Dv90:	L D Dv90: 240	L D Dv90: 240	API
diffraction)		L D Dv90:	240 micron	micron	micron
Manufacturing	API size	API size	API size	API size	API size	API size
process	reduction	reduction by	reduction by	reduction by	reduction by	dispersed
	by HSM	HSM	HSM (F001e),	HSM followed	HSM followed	by HSM
			then	by HPH, then	by HPH, then	and
			subsequent	subsequent	subsequent	sonication/
			dextrose	dextrose	NaCl addition	vortex
			addition	addition
Process End	L D Dv90:	L D Dv90:	L D Dv90:	Total 15 min	Total 15 min	L D Dv90:
Point Targets	14.7	17.6 micron	13.6 micron	HSM at	HSM at	12.2
(PSD via laser	micron	and	and L D Dv50:	~25,000 rpm to	~25,000 rpm to	micron
diffraction)	and	L D Dv50: 6.9	5.7 micron	target	target	HSM:
	L D Dv50:	micron	HSM: Total	L D Dv90:<~40	L D Dv90:<~40	Total 5 min
	5.9	HSM: Total	60-120 min	micron	micron	at
	micron	>120 min at	at	L D Dv90: 15.1	L D DV90: 15.1	~25,000 rpm
	HSM:	~25,000-30,000 rpm	~25,000 rpm	micron	micron	to
	Total		before	HPH: 15 passes	HPH: 15 passes	L D Dv90:
	>120 min		dextrose	in parallel flow	in parallel flow	12.6
	at		addition to	through z5	through z5	micron by
	~25,000-30,000 rpm		L D Dv90: 13.0	nozzle at	nozzle at	sonication/
			micron	~40,000 psi	~40,000 psi	vortex
				before dextrose	before NaCl
				addition to	addition to
				L D Dv90: 12.1	L D Dv90:
				micron	12.2 micron
				pH = 7.3	pH = 7.5

	TABLE 25

	Target Formulation (mg/mL)

	F003k2	F003k3	F003l	F005a2

Fulvestrant

	100	100	100	100
Polysorbate 80	5	5	5	5
Dextrose	50	50	50	—
Mannitol	—	—	—	50
WFI	q.s. to	q.s. to	q.s. to volume	q.s. to volume
Starting API	Jet-mill API	Jet-mill API	Jet-mill API L D Dv90:	Un-milled API L D Dv90:
(PSD via laser	L D Dv90: 7-8	L D Dv90: 7-8	7-13 micron	240 micron
diffraction)	micron	micron
Manufacturing	API dispersed	API dispersed by	API dispersed by HSM	API size reduction by
process	by HSM, then	HSM then size	(F003i, f, j). Suspension	HSM (F001e), then
	subsequent	reduction by HPH	lyophilized, reconstituted	mannitol addition.
	dextrose	in the presence of	then composited	Suspension lyophilized,
	addition	dextrose		reconstituted then
				composited
Process End	L D Dv90: 6.7	Total 5 min HSM	Total	5 min HSM at	L D Dv90: 13.6 micron
Point Targets	micron after	at	~25,000 rpm for	and L D Dv50: 5.7 micron
(PSD via laser	dextrose	~25,000 rpm	dispersing each	HSM: Total 60-120 min.
diffraction)	addition	(Dv90: 9.2	individual API lot	at
	HSM: Total 5 min	micron) for	F003i, f, j before	~25,000 rpm before
	at	dispersing API	lyophilization, L D Dv90:	mannitol addition to
	~25,000 rpm	L D Dv90: 13.3	7.0, 12.9, 7.3 micron	L D Dv90: 13.2 micron
		micron HPH: 9	After reconstitution,	(F005a)
		passes in parallel	L D Dv90: 121, 113, 113	After lyophilization,
		flow through z5	micron.	reconstitution, composite
		nozzle at	Composite F003l,	F005a2, Dv90: 18.4
		~40,000 psi;	L D Dv90: 112 micron	micron
		dextrose co-
		processed

	TABLE 26

	Target Formulation (mg/mL)

	F005b1	F005c2	F005c3	F005d1	F015a1	F015a3

Fulvestrant	100	100	100	100	100	100
Polysorbate 80	5	5	5	5	25	25
Mannitol	50	50	50	50	50	50
WFI	q.s. to	q.s. to	q.s. to	q.s to	q.s. to	q.s. to
Starting API	Jet-mill API	Jet-mill	Jet-mill API	Jet-mill API	Jet-mill API	Jet-mill API
(PSD via laser	L D Dv90: 7-8	API	L D Dv90: 7-8	L D Dv90: 7-8	L D Dv90: 7-8	L D Dv90: 7-8
diffraction)	micron	L D Dv90:	micron	micron	micron	micron
		7-8 micron
Manufacturing	API	API	Lyophilized	API size	API	Lyophilized
process	dispersed by	dispersed	F005c2	reduction by	dispersed by	F015a1
	HSM in the	by HSM		HSM followed	HSM in the
	presence of	then size		by HPH, then	presence of
	mannitol	reduction		subsequent	mannitol
		by HPH in		mannitol
		the		addition
		presence of
		mannitol
Process End	L D Dv90:	Total 5 min	After	To total 5 min	L D Dv90:	After
Point Targets	7.9	HSM at	reconstitution,	HSM at	6.9	reconstitution,
(PSD via laser	micron	~25,000 rpm	L D Dv90:	~25,000 rpm for	micron	L D Dv90:
diffraction)	HSM: Total	for	112 micron	dispersing API	HSM: Total	22.7
	5 min at	dispersing		L D Dv90: ~7	5 min at	micron
	~25,000 rpm	API		micron	~25,000 rpm
		(L D Dv90:		HPH with 15
		~7 micron)		passes in parallel
		To HPH		flow through z5
		with 15		nozzle at
		passes in		~40,000 psi
		parallel		resulted in
		flow		Dv90:
		through z5		~16 micron
		nozzle at		(F001h4)
		~40,000 psi		L D Dv90: 4.0
		resulted in		micron after
		L D Dv90:		mannitol
		17.2 micron		addition
		pH = 7.1		pH = 7.1

	TABLE 27

	Target Formulation (mg/mL)

	F015a4	F017a1	F017a3	F005g4	F005g5

Fulvestrant

	100	100	100	100	100
Polysorbate 80	25	15	15	5	5
Mannitol	50	50	50	50	50
WFI	q.s. to	q.s. to	q.s. to	q.s. to	q.s. to
Starting API	Jet-mill API	Jet-mill API	Jet-mill API	Recrystallized API	Recrystallized
(PSD via laser	L D Dv90: 7-8	L D Dv90: 7-8	L D Dv90: 7-8	L D Dv90: 18	API
diffraction)	micron	micron	micron	micron	L D Dv90: 18
					micron
Manufacturing	F015a3 gamma	API dispersed by	Lyophilized,	API dispersed by	Lyophilized
process	irradiated at 35	HSM in the	gamma	HSM in the	F005g4
	kGy	presence of	irradiated (35	presence of
		mannitol	KGy) F017a1	mannitol
Process End	After	L D Dv90: 7.2	After	Total 5 min HSM at
Point Targets	reconstitution,	micron	reconstitution,	~25,000 rpm for
(PSD via laser	L D Dv90: 22.7	HSM: Total 5 min	L D Dv90: 31.9	dispersing API
diffraction)	micron	at	micron	(L D Dv90: ~20
		~25,000 rpm		micron)
				L D Dv90: ~10
				micron after HPH
				with 9 passes in
				parallel flow
				through z5 nozzle at
				~40,000 psi
				Final L D Dv90: 7.5
				micron after
				concentration

D. Pharmacokinetics

In certain embodiments of the invention, the pharmaceutical compositions are bioequivalent to the commercial pharmaceutical composition, FASLODEX™. The single dose PK parameters in postmenopausal advanced breast cancer patients administered FASLODEX™ dosed intramuscularly with 500 mg with an additional dose at day 15 are reported as, in geometric mean and coefficient of variation (%), Cmax 25.1 (35.3) ng/mL, Cmin 16.3 (25.9) ng/mL, and AUC 11,400 (33.4) ng·hr/mL.
In further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™. In yet further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™ in the fasting state. In still further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™ in the fed state.
In other embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention having a fulvestrant concentration of 100 mg/mL is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™. In still other embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention having a fulvestrant concentration of 100 mg/mL is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™ in the fasting state. In yet other embodiments of the invention, the 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)of the pharmaceutical composition of the invention having a fulvestrant concentration of 100 mg/mL is within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of FASLODEX™ in the fed state.
In particular embodiments of the invention, the pharmaceutical composition has the single dose and multiple dose pharmacokinetic parameters shown in Tables 28 and 29. Table 28 shows pharmacokinetic parameters for 500 mg dosage of pharmaceutical compositions of the disclosure. For the data labeled “Single Dose” in Table 28, the fulvestrant blood plasma concentration data are shown for a 500 mg initial dose with an additional 500 mg dose given on day 15. For the data labeled “Multiple Dose Steady State” in Table 28, the fulvestrant blood plasma concentration data are shown for measurement at month 3, after a 500 mg dosage on days 1, 15, 20, and once monthly thereafter. Table 29 shows pharmacokinetic parameters for a single 250 mg dosage of pharmaceutical compositions of the disclosure. In Table 29, data are expressed as geometric mean (CV %), except for T_max, which is shown as a median value with a range indicated in parentheses.

TABLE 28

C_max(ng/mL)	C_min(ng/mL)	AUC (ng · hr/mL)

Single Dose¹	20.08-31.375	13.04-20.375	9,120-14,250
Multiple Dose	22.4-35.0	9.76-15.25	10,480-16,375
Steady State²

TABLE 29

I	II	III	IV	V	VI	VII

C_max	8.20	4.76	8.2	4-8.5	11.8	8.3	8-12
(μg/L)	(63.8)	(68.1)			(6.6)	(8.8)
C_min	2.62	2.38	2.6	2.0-3.0
(μg/L)	(33.4)	(47.7)
T_max	6.97	8.8	7	6-9	4.2	4.6	4-5
(days)	(1.86-7.95)	(6.97-12.0)			(8.3)	(11.2)
AUC₂₈	148	88.4	148	80-150	369	333	325-375
(μg · day/L)	(45.3)	(47.3)			(4.1)	(3.0)

In particular embodiments, a dose of about 500 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™. In certain embodiments, a dose of less than 500 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™. In further embodiments, a dose of about 400 to 450 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™. In still further embodiments, a dose of about 350 to 400 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™. In yet further embodiments, a dose of about 300 to 350 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™. In even further embodiments, a dose of about 250 to 300 mg of a fulvestrant pharmaceutical composition of the invention is bioequivalent to 500 mg of the commercial pharmaceutical composition, FASLODEX™.
In other embodiments of the invention, a 500 mg dose of a pharmaceutical composition of the invention provides 90% confidence intervals (CI) of the relative mean C_max, AUC_(0-t)and AUC_(0-∞)within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of a 500 mg dose of FASLODEX™.
In other embodiments of the invention, a dose of less than 500 mg of a pharmaceutical composition of the invention provides 90% confidence intervals (CI) of the relative mean C_ma, AUC_(0-t)and AUC_(0-∞)within 80% to 125% of the relative mean C_max, AUC_(0-t)and AUC_(0-∞), respectively, of a 500 mg dose of FASLODEX™.
In some embodiments of the invention, fulvestrant pharmaceutical compositions of the invention can be administered as a single intramuscular injection, with the 90% confidence intervals (CI) of the relative mean Cmax, AUC(0-t) and AUC(0-∞) of fulvestrant is within 80% to 125% of the relative mean Cmax, AUC(0-t) and AUC(0-∞), respectively, of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections. In further embodiments, such fulvestrant pharmaceutical compositions administered as a single intramuscular injection comprise a dose of about 500 mg of fulvestrant. In yet further embodiments, such fulvestrant pharmaceutical compositions administered as a single intramuscular injection comprise a dose of about 500 mg of fulvestrant in an injection volume of about 3.0 mL to about 5.0 mL, about 3.5 mL to about 4.5 mL, or about 4.0 mL.
In certain embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is less than 80% of the relative mean Cmax of FASLODEX™. It is believed that such embodiments may provide benefits by providing a therapeutically effect amount of fulvestrant exposure to a subject while reducing the degree of one or more Cmax-driven side-effects or toxicities in comparison to the degree of side-effects or toxicities experienced by a subject from receiving a therapeutically effective amount of fulvestrant exposure from one or more dosages of FASLODEX™.
In some embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, or less than 40% of the relative mean Cmax of FASLODEX™. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.
In yet further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, or less than 40% of the relative mean Cmax of FASLODEX™ in the fasting state. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.
In still further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, less than 50%, less than 45%, or less than 40% of the relative mean Cmax of FASLODEX™ in the fed state. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.
In some embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, within about 45% to about 55%, within about 55% to about 65%, within about 65% to about 75%, within about 50% to about 60%, within about 60% to about 70%, or within about 70% to about 80% of the relative mean Cmax of FASLODEX™. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.
In yet further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, within about 45% to about 55%, within about 55% to about 65%, within about 65% to about 75%, within about 50% to about 60%, within about 60% to about 70%, or within about 70% to about 80% of the relative mean Cmax of FASLODEX™ in the fasting state. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.
In still further embodiments of the invention, the 90% confidence intervals (CI) of the relative mean AUC_(0-t), relative mean AUC_(0-∞), or both of fulvestrant pharmaceutical compositions of the invention is within 80% to 125% of the relative mean AUC_(0-t)and relative mean AUC_(0-∞), respectively, of FASLODEX™, and the relative mean Cmax of fulvestrant pharmaceutical compositions of the invention is about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, within about 45% to about 55%, within about 55% to about 65%, within about 65% to about 75%, within about 50% to about 60%, within about 60% to about 70%, or within about 70% to about 80% of the relative mean Cmax of FASLODEX™ in the fed state. In further embodiments, such fulvestrant pharmaceutical compositions are administered as a single intramuscular injection and comprise a dose of about 500 mg of fulvestrant at a concentration of about 100 mg/mL.

E. Methods of Treatment

In further embodiments, the invention is directed to methods of treatment comprising administration of a pharmaceutically effective amount of any of the fulvestrant pharmaceutical compositions described herein to a patient in need thereof. In particular embodiments, the invention is directed to a method of treating breast cancer, comprising administering a pharmaceutically acceptable amount of any of the fulvestrant pharmaceutical compositions described herein. In certain embodiments, the breast cancer is metastatic breast cancer. In other embodiments of the invention, the breast cancer is hormone receptor (HR)-positive breast cancer. In still other embodiments of the invention, the invention is directed to a method of treating hormone receptor (HR)-positive breast cancer in a post-menopausal woman comprising administration of a pharmaceutically effective amount of any of the fulvestrant pharmaceutical compositions described herein. In yet other embodiments, the invention is directed to a method of treating hormone receptor (HR)-positive breast cancer in a post-menopausal woman with disease progression following antiestrogen therapy comprising administration of a pharmaceutically effective amount of any of the fulvestrant pharmaceutical compositions described herein. In yet further embodiments, the invention is directed to a method of treating HR-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer in a woman with disease progression after endocrine therapy.
In particular embodiments of the invention, a fulvestrant pharmaceutical composition as described herein is administered on days 1, 15, 29, and once monthly thereafter. In further embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered on days 1, 15, 29, and once monthly thereafter. In still further embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered on days 1, 15, 29, and once monthly thereafter.
In certain embodiments of the invention, a fulvestrant pharmaceutical composition as described herein is administered as a single injection. In other embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single injection. In yet other embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single 5 mL injection. In further embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single 4 mL injection. In yet further embodiments, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single 3 mL injection. In still other embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single injection. In further embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single 2.5 mL injection. In yet further embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as a single 5 mL injection.
In particular embodiments of the invention, a fulvestrant pharmaceutical composition as described herein is administered as two injections. In further embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as two injections. In still further embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as two 5 mL injections. In yet further embodiments of the invention, a 500 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as two 2 mL injections, two 2.5 mL injections, two 3 mL injections, two 3.5 mL injections, or two 4 mL injections. In other embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as two injections. In yet other embodiments of the invention, a 250 mg dose of any of the fulvestrant pharmaceutical compositions as described herein is administered as two 2.5 mL injections.
The fulvestrant pharmaceutical compositions described herein may be administered alone, or in combination with one or more additional therapeutic agents as defined herein. An additional therapeutic agent may be used to treat one or more core symptoms and/or comorbidities associated with cancer in general or breast cancer in particular. In one aspect, fulvestrant is formulated (and administered) with at least one therapeutic agent as a fixed dose. In another aspect, fulvestrant is formulated (and administered) separately from the therapeutic agent(s).
Some examples of therapeutic agents that may be used in combination with fulvestrant include, but are not limited to, e.g., a EGFR kinase inhibitor, a PDGFR kinase inhibitor, a FGFR kinase inhibitor, or any of the other cytotoxic, chemotherapeutic, antihormonal, anti-angiogenic, antiproliferative, pro-apoptotic, anti-HER2, radiation or a radiopharmaceutical, signal transduction inhibitors, or other anti-cancer agents or treatments. Examples of particular agents that can be used in combination with the fulvestrant pharmaceutical compositions of the disclosure include palbociclib, letrozole, anastrozole, doxorubicin, paclitaxel, docetaxel, vinorelbine, and 5-fluorouracil. In other embodiments, therapeutic agents that may be used in combination with fulvestrant include, but are not limited to, agents or treatments for one or more of pain, nausea, emesis, hot flushes, constipation, and dizziness.
Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred embodiments of the disclosure and that such changes and modifications can be made without departing from the spirit of the disclosure. It is, therefore, intended that the following examples and appended claims cover all such equivalent variations as fall within the true spirit and scope of the disclosure.
The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety.

EXAMPLES

Example: Preparation of Fulvestrant Pharmaceutical Compositions/Variants

Some exemplary fulvestrant pharmaceutical compositions were prepared with 50 mg/mL and 100 mg/mL concentrations of fulvestrant in aqueous suspensions. Tables 1-20 show aspects of the pharmaceutical compositions of the pharmaceutical compositions and the methods of preparation of some of the pharmaceutical compositions, also referred to as formulations, variants, or Lots in the Tables. Tables 4-7 and 23-27 and FIGS. 4-12 and 15 show aspects of the methods of preparation used to prepare some of the pharmaceutical compositions.
Where indicated in the Tables and Figures, the formulations B, E, I, J, K, L, L3F, L6, F003a, F003b, F003e, F004a, F003k2, F003k3, F005a2, F003l, F005b1, F015a1, F015a3, F005d1, F005c3, F005g5 tested in Studies 1-3 below were prepared via one or more of process steps of (1) low shear mixing, indicated in the Tables 23-27 and FIGS. 4-12 as “Mix” or “Mix with Vortex Mixer” steps; (2) high shear mixing, indicated as “HSM” or “Homogenize” steps; (3) high pressure homogenization, indicated as “HPH” or “Process with Nano DeBee” steps; (4) concentration via supernatant removal; and (5) application of sonication. Where indicated, supernatant removal was performed by phase separating the pharmaceutical composition and withdrawing the desired amount of supernatant to concentrate the suspensions to the target concentrations of fulvestrant, either 50 mg/mL or 100 mg/mL, depending on the pharmaceutical composition. Where indicated, phase separation was performed by overnight settling in a clear glass centrifuge tube. Application of centrifuge for phase separation could also be utilized.
Fulvestrant active pharmaceutical ingredient (which may also be referred to as “API” herein and in the Tables and Figures) was obtained in un-milled forms or in milled, micronized, or recrystallized forms from commercial suppliers. As-obtained particle size distributions for un-milled API varied from an LD Dv(90) of about 240 microns to an LD Dv(90) of about 2130 microns. As-obtained particle size distributions for milled, micronized, and recrystallized API varied from an LD Dv(90) of about 7 microns to an LD Dv(90) of about 18 microns. Fulvestrant API may be obtained in various particle size distributions from commercial sources and processed as described elsewhere herein to achieve the desired particle size distributions. Particle size distributions can be monitored throughout the processing steps through analysis of samples as described elsewhere herein.
Where indicated in the Tables and Figures, the formulations B, E, I, J, K, L, L3F, L6, F003a, F003b, F003e, F004a, F003k2, F003k3, F005a2, F003l, F005b1, F015a1, F015a3, F005d1, F005c3, F005g5 tested in Studies 1-3 below were prepared via high shear mixing (HSM) steps. The preparation of formulations can be performed with an IKA T10 Basic Disperser with an IKA S10N-10G dispersing tool. At the speeds indicated (˜20,000 to 30,000 rpm), the mixture of fulvestrant and suspension vehicle was processed in cycles until the total processing time indicated was reached. Between each cycle, a formulation was vortexed at ˜3000 rpm for 30 seconds then sonicated for 1 minute to remove or reduce foam generated during the high shear mixing by the disperser. Formulations were also rested as needed in between cycles at room temperature to allow the disperser to cool down and avoid overheating of the product and the equipment. Sonication was performed with a Branson 3800 Ultrasonic Bath (Branson Ultrasonics Corp., Danbury, Conn.) at a frequency of 40 kHz. Other mixing and ultrasonic apparatuses may also be used to achieve mixing and particle size distribution as desired.
In some embodiments, high pressure homogenization was performed. In certain embodiments, high pressure homogenization (HPH) steps were performed with a Nano DeBEE High Pressure Homogenizer (BEE International, South Easton, Mass.) in a Labconco XPert Filtered Balance System (Model 3950630) (Labconco, Kans. City, Mo.), installed the 100 ml sample holder and Z5 nozzle in parallel flow configuration on Nano DeBEE High Pressure Homogenizer. The homogenizer was primed with filing water until the process pressure reached the processing pressure as indicated in the Tables and Figures. Water was removed from the system using the plunger to minimize the dilution of the batch by the residual priming water. Approximately ˜50 ml of the suspension for HPH processing was loaded from the 50 mL clear Pyrex glass bottle on Nano DeBEE High Pressure Homogenizer. The Nano DeBEE was run in continuous mode until the pressure reached the indicated target processing pressure. The suspension was then processed for the indicated number passes at the processing pressure. To avoid losing the prime of the system and consequently the processing pressure, only total ˜40 mL (8 strokes of ˜5 mL per stroke) of the suspension was processed and collected from each pass. The 40 mL suspension was then loaded back to the reservoir for the suspension to be processed in the next pass. After the processing was completed, 40 ml fine suspension was collected in a 100 mL clear Pyrex glass bottle by running Nano DeBEE High Pressure Homogenizer until no sample was pumped out. In certain embodiments, high pressure homogenization was performed with other apparatuses at processing pressures ranging from about 5,000 psi to about 45,000 psi. Other high pressure homogenization apparatuses may also be used to achieve the desired particle size distributions described herein.
Some formulations for Study 3 below were lyophilized and reconstituted with sterile water for injection, USP prior to administration, as indicated in the Example below.
References to “Assay” refers to high-performance liquid chromatography (HPLC) measurement of the fulvestrant concentration of the pharmaceutical composition at intermediate processing steps or in final result as prepared. The “Assay” results are given in absolute measured mg/mL or as a percentage (%) or (% LC), where percentages indicate the concentration of fulvestrant relative to the 50 mg/mL label claim of the commercially available FASLODEX™ product. Total impurities were also measured and are provided in the figures as a percentage by area (% a/a) where indicated. HPLC was performed with Agilent Technologies Agilent 1260 Infinity Quaternary LC module G1311B (Agilent Technologies, Santa Clara, Calif.). Other HPLC apparatuses may also be used to analyze the fulvestrant concentrations.
In some aspects, particle size and particle size distributions were analyzed with Malvern Mastersizer 3000 (Malvern Instruments Ltd., Malvern, Worcestershire, UK), with an attached sample dispersion unit with an in-line sonication probe for agglomerate dispersion prior to analysis via laser diffraction.
In some aspects, particle size and particle size distributions were analyzed with Malvern Morphologi G3 (Malvern Instruments Ltd., Malvern, Worcestershire, UK), to determine circle equivalent (CE) diameters via microscopy image capture and analysis.
Measurements of pH were obtained at ambient room temperature with a Thermo Scientific Orion Star A211 pH Meter (Thermo Fisher Scientific Inc., Waltham, Mass.).

Example: Pharmacokinetic Study 1 of Intramuscular Administration to Female Dogs

Fulvestrant pharmaceutical compositions B, E, I, J, K, and L were prepared as described elsewhere herein and in the Figures. A preclinical study was performed to determine the pharmacokinetics of the pharmaceutical compositions following a single intramuscular administration of 15.4 mg/kg to female dogs. The pharmacokinetics of 15.4 mg/kg IM FASLODEX™ (fulvestrant injection, 250 mg/5 mL) were also determined and used for comparison to the three prototype pharmaceutical compositions. The 15.4 mg/kg dose used in this study is the canine equivalent, in mg/m², of the maximum dose (500 mg) for human use and was scaled for use in canine by dividing the dose (based on a 60 kg human) by a canine species conversion factor of 0.54.
Twenty-four non-naïve female beagle dogs were used in the study. The animals weighed between approximately 5-10 kg. Animal welfare for this study was in compliance with the U.S. Department of Agriculture's (USDA) Animal Welfare Act (9 Code of Federal Regulations (CFR) Parts 1, 2 and 3). The Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Academy Press, Washington, D.C., was followed. The facility maintained an Animal Welfare Assurance statement with the National Institutes of Health, Office of Laboratory Animal Welfare.
The FASLODEX™ test articles contained a small molecule that was used as received and no adjustment was made for purity, salt correction, etc. The FASLODEX™ test articles were gently agitated prior to dispensing and dose delivery. Pharmaceutical Composition B, Pharmaceutical Composition E, Pharmaceutical Composition I, Pharmaceutical Composition J, Pharmaceutical Composition K, and Pharmaceutical Composition L were stored at room temperature and protected from light prior to use, and gently agitated prior to dispensing and dose delivery.
The animals were not fasted prior to dosing. Each animal received a single intramuscular (IM) dose of only one of the appropriate test article pharmaceutical compositions as outlined in the following study design table, Table 30. IM doses were administered with a 20 G needle via bolus injection into the same large muscle mass (using the Z-track injection technique) in the left hind limb of each animal. Attempts were made for consistent injections between animals [selection of the dose site (muscle), depth, etc.]. The hair was clipped from the injection site prior to dosing. The injection site was marked following dosing and remarked as necessary throughout the study. Specifications for all dose delivery were recorded and reported in the study report [including, but not limited to needle gauge/length, syringe size/barrel type with manufacturer and part number, estimated injection depth into the muscle, approximate duration required to administer the injection; any substantial resistance (either flow through the syringe/needle and/or into the muscle during administration)] was documented.

TABLE 30

			Pharmaceutical
			Composition
			Fulvestrant	Dose	Dose
		No. of	Concentration	Level	Volume
Group	Test Article	Females	(mg/mL)	(mg/kg)	(mL/kg)

1	FASLODEX ™	3	50	15.4	0.308
2	Pharmaceutical	3	50	15.4	0.308
	Composition B
3	Pharmaceutical	3	50	15.4	0.308
	Composition E
4	FASLODEX ™	3	50	15.4	0.308
5	Pharmaceutical	3	50	15.4	0.308
	Composition I
6	Pharmaceutical	3	50	15.4	0.308
	Composition J
7	Pharmaceutical	3	50	15.4	0.308
	Composition K
8	Pharmaceutical	3	100	15.4	0.154
	Composition L

All animals were observed at least twice a day for morbidity, mortality, injury, and availability of food and water. Any animals in poor health were identified for further monitoring and possible euthanasia.
Blood samples were collected at various time intervals to measure the blood plasma concentration of fulvestrant. Blood samples for Groups 1-3 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, and 12 (on Day 1); and 24 (on Day 2), 48 (on Day 3), 120 (on Day 6), 192 (on Day 9), 264 (on Day 12), 336 (on Day 15), 384 (on Day 17), 456 (on Day 20), 528 (on Day 23), 600 (on Day 26), and 672 (on Day 29) hours postdose. Blood samples for Groups 4-8 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, and 12 (on Day 1); and 24 (on Day 2), 48 (on Day 3), 72 (on Day 4), 96 (on Day 5), 120 (on Day 6), 192 (on Day 9), 264 (on Day 12), 336 (on Day 15), 384 (on Day 17), 528 (on Day 23), and 672 (on Day 29) hours postdose.
Whole venous blood samples of approximately 2 mL each were collected from a peripheral vein of all animals for determination of fulvestrant exposure. Blood was collected with sodium heparin anticoagulant (glass tube, no gel separator). All blood samples were placed on wet ice following collection until centrifuged. Blood was centrifuged at 3500 rpm for 7 minutes at 2 to 8° C. Plasma (minimum of 0.8 mL volume) was separated from blood cells within 0.75 hours of blood collection and frozen. Plasma samples were initially placed on dry ice prior to being stored in the appropriate freezer (−60 to −90° C.). Samples were shipped on dry ice for bioanalytical analysis.
A model independent method was used to determine C_maxand AUC values from fulvestrant plasma concentration-time data. Results are shown in Tables 31-37 and FIGS. 1B, 2A, 2C, and 3. Table 31 shows the pharmacokinetic data from the 15.4 mg/kg dosages as nominally dosed (based on the target fulvestrant concentration for each pharmaceutical composition). An “Assay %” is shown for the fulvestrant pharmaceutical compositions of the present disclosure used in the study. The “Assay %” represents the percentage equivalence of the particular pharmaceutical composition in comparison to the FASLODEX™ label claim fulvestrant concentration, with “Assay %” values determined via HPLC, measurement samples taken pre- and post-dose, with one value selected for normalization. The data in Tables 32-37 are normalized using the “Assay %” values to compare PK results based upon the actual mg/kg of fulvestrant administered, assuming linear scaling. FIGS. 1B, 2A, 2C, and 3 depict graphs of the dose normalized fulvestrant mean plasma concentrations.

TABLE 31

PK parameters based upon the nominal dose 15.4 mg/kg

	Assay (%)	Assay (%)
	(measured	(measured
Variant	pre-dose)	post-dose)

(Geometric Mean of		AUC0-14 d	AUC0-28 d	Starred value used for
n as indicated)	Cmax (ng/mL)	(hr * ng/mL)	(hr * ng/mL)	normalization

Faslodex	35.0	7015	8917
(LW466, n = 9)
Faslodex	45.7	7666	9306
(MB122, n = 6)
Faslodex	32.0	7177	9018
(MB948, n = 3)
Faslodex	36.9	8149	9817
(MC949, n = 4)
Faslodex	37.5	7408	9195
(All, n = 22)
B (n = 3)	8.7	1930	3250	100.0*	138.9
E (n = 3)	29.1	5750	8380	93.4*	91.8
I (n = 3)	41.5	8840	12300	95.6*	99.6
J (n = 3)	44.8	5750	8100	87.6*	93.6
K (n = 3)	69.7	7540	9630	84.6*	91.2
L (n = 3)	63.9	8430	11000	94.1*	97.3
L3F (n = 3)	22.6	5140	7130	83.8*	79.0
L6 (n = 3)	24.7	6050	9360	113.9*	113.9
F003a (n = 3)	27.7	5860	8610	95.0*	97.1
F003b (n = 3)	32.5	7210	9650	96.3*	97.4
F003e (n = 3)	28.5	6400	9080	100.6*	100.5
F004a (n = 3)	31.7	4310	6190	99.8*	100.4
F003k2 (n = 4)	33	3910	5960	97.8*	100.1
F003k3 (n = 4)	26.8	5430	7060	99.6*	100.7
F005a2 (L, n = 3)	19.7	4370	6840	—	93.2*
F003l (L, n = 3)	25.1	5510	8680	—	99.2*
F005b1 (n = 3)	49.7	9420	12100	100.2*	—
F015a1 (n = 3)	63.6	4750	7120	103.8*	—
F015a3 (L, n = 3)	34.4	3850	6000	—	98.6*
F005d1 (n = 3)	37.9	7180	9910	100.5*	99.2
F005c3 (L, n = 3)	17.7	3680	5820	95.2*	—

TABLE 32

PK parameters normalized against the actual dose

	Cmax
Variant (Geometric	(ng/mL	AUC0-14 d	AUC0-28 d
Mean of n as	per	(hr * ng/mL per	(hr * ng/mL per
indicated)	mg/kg)	mg/kg)	mg/kg)

Faslodex (LW466,	2.3	456	579
n = 9)
Faslodex (MB122,	3.0	498	604
n = 6)
Faslodex (MB948,	2.1	466	586
n = 3)
Faslodex (MC949,	2.4	529	637
n = 4)
Faslodex	2.4	481	597
(All, n = 22)
B (n = 3)	0.6	125	211
E (n = 3)	2.0	400	583
I (n = 3)	2.8	600	835
J (n = 3)	3.3	426	600
K (n = 3)	5.3	579	739
L (n = 3)	4.4	582	759
L3F (n = 3)	1.8	398	552
L6 (n = 3)	1.4	345	534
F003a (n = 3)	1.9	401	589
F003b (n = 3)	2.2	486	651
F003e (n = 3)	1.8	413	586
F004a (n = 3)	2.1	280	403
F003k2 (n = 4)	2.2	260	396
F003k3 (n = 4)	1.7	354	460
F005a2 (L, n = 3)	1.4	304	477
F003l (L, n = 3)	1.6	361	568
F005b1 (n = 3)	3.2	610	784
F015a1 (n = 3)	4.0	297	445
F015a3 (L, n = 3)	2.3	254	395
F005d1 (n = 3)	2.4	464	640
F005c3 (L, n = 3)	1.2	251	397

TABLE 33

Comparison of normalized PK parameters against all Faslodex lots

Variant	Cmax
(Geometric Mean of	ratio to	AUC0-14 d ratio	AUC0-28 d ratio
n as indicated)	Faslodex (%)	to Faslodex (%)	to Faslodex (%)

Faslodex (LW466,	93	95	97
n = 9)
Faslodex (MB122,	122	103	101
n = 6)
Faslodex (MB948,	85	97	98
n = 3)
Faslodex (MC949,	98	110	107
n = 4)
Faslodex	100	100	100
(All, n = 22)
B (n = 3)	23	26	35
E (n = 3)	83	83	98
I (n = 3)	116	125	140
J (n = 3)	136	89	101
K (n = 3)	219	120	124
L (n = 3)	181	121	127
L3F (n = 3)	72	83	93
L6 (n = 3)	58	72	89
F003a (n = 3)	78	83	99
F003b (n = 3)	90	101	109
F003e (n = 3)	75	86	98
F004a (n = 3)	85	58	67
F003k2 (n = 4)	90	54	66
F003k3 (n = 4)	72	74	77
F005a2 (L, n = 3)	56	63	80
F003l (L, n = 3)	67	75	95
F005b1 (n = 3)	132	127	131
F015a1 (n = 3)	163	62	75
F015a3 (L, n = 3)	93	53	66
F005d1 (n = 3)	100	96	107
F005c3 (L, n = 3)	50	52	66

TABLE 34

Comparison of normalized PK parameters against Faslodex lot
LW466

	Cmax
Variant	ratio to
(Geometric Mean of n	Faslodex	AUC0-14 d ratio	AUC0-28 d ratio
as indicated)	(%)	to Faslodex (%)	to Faslodex (%)

Faslodex (LW466,	100	100	100
n = 9)
Faslodex (MB122,	131	109	104
n = 6)
Faslodex (MB948,	92	102	101
n = 3)
Faslodex (MC949,	106	116	110
n = 4)
Faslodex	107	106	103
(All, n = 22)
B (n = 3)	25	28	36
E (n = 3)	89	88	101
I (n = 3)	124	132	144
J (n = 3)	146	94	104
K (n = 3)	236	127	128
L (n = 3)	194	128	131
L3F (n = 3)	77	87	95
L6 (n = 3)	62	76	92
F003a (n = 3)	83	88	102
F003b (n = 3)	97	107	112
F003e (n = 3)	81	91	101
F004a (n = 3)	91	62	70
F003k2 (n = 4)	97	57	68
F003k3 (n = 4)	77	78	79
F005a2 (L, n = 3)	60	67	82
F003l (L, n = 3)	72	79	98
F005b1 (n = 3)	142	134	135
F015a1 (n = 3)	175	65	77
F015a3 (L, n = 3)	100	56	68
F005d1 (n = 3)	108	102	111
F005c3 (L, n = 3)	53	55	69

TABLE 35

Comparison of normalized PK parameters against Faslodex lot
MB122

Faslodex (LW466,	76	92	96
n = 9)
Faslodex (MB122,	100	100	100
n = 6)
Faslodex (MB948,	70	94	97
n = 3)
Faslodex (MC949,	81	106	105
n = 4)
Faslodex	82	97	99
(All, n = 22)
B (n = 3)	19	25	35
E (n = 3)	68	80	96
I (n = 3)	95	121	138
J (n = 3)	112	86	99
K (n = 3)	180	116	122
L (n = 3)	148	117	126
L3F (n = 3)	59	80	91
L6 (n = 3)	47	69	88
F003a (n = 3)	64	80	97
F003b (n = 3)	74	98	108
F003e (n = 3)	62	83	97
F004a (n = 3)	69	56	67
F003k2 (n = 4)	74	52	65
F003k3 (n = 4)	59	71	76
F005a2 (L, n = 3)	46	61	79
F003l (L, n = 3)	55	72	94
F005b1 (n = 3)	108	123	130
F015a1 (n = 3)	134	60	74
F015a3 (L, n = 3)	76	51	65
F005d1 (n = 3)	82	93	106
F005c3 (L, n = 3)	41	50	66

TABLE 36

Comparison of normalized PK parameters against Faslodex lot
MB948

Faslodex (LW466,	109	98	99
n = 9)
Faslodex (MB122,	143	107	103
n = 6)
Faslodex (MB948,	100	100	100
n = 3)
Faslodex (MC949,	115	114	109
n = 4)
Faslodex	117	103	102
(All, n = 22)
B (n = 3)	27	27	36
E (n = 3)	97	86	99
I (n = 3)	136	129	143
J (n = 3)	160	91	103
K (n = 3)	257	124	126
L (n = 3)	212	125	130
L3F (n = 3)	84	85	94
L6 (n = 3)	68	74	91
F003a (n = 3)	91	86	100
F003b (n = 3)	105	104	111
F003e (n = 3)	89	89	100
F004a (n = 3)	99	60	69
F003k2 (n = 4)	105	56	68
F003k3 (n = 4)	84	76	79
F005a2 (L, n = 3)	66	65	81
F003l (L, n = 3)	79	77	97
F005b1 (n = 3)	155	131	134
F015a1 (n = 3)	191	64	76
F015a3 (L, n = 3)	109	54	67
F005d1 (n = 3)	118	100	109
F005c3 (L, n = 3)	58	54	68

TABLE 37

Comparison of normalized PK parameters against Faslodex lot
MB949

Faslodex (LW466,	95	86	91
n = 9)
Faslodex (MB122,	124	94	95
n = 6)
Faslodex (MB948,	87	88	92
n = 3)
Faslodex (MC949,	100	100	100
n = 4)
Faslodex	102	91	94
(All, n = 22)
B (n = 3)	24	24	33
E (n = 3)	84	76	91
I (n = 3)	118	113	131
J (n = 3)	139	81	94
K (n = 3)	223	109	116
L (n = 3)	184	110	119
L3F (n = 3)	73	75	87
L6 (n = 3)	59	65	84
F003a (n = 3)	79	76	92
F003b (n = 3)	91	92	102
F003e (n = 3)	77	78	92
F004a (n = 3)	86	53	63
F003k2 (n = 4)	91	49	62
F003k3 (n = 4)	73	67	72
F005a2 (L, n = 3)	57	58	75
F003l (L, n = 3)	69	68	89
F005b1 (n = 3)	134	115	123
F015a1 (n = 3)	166	56	70
F015a3 (L, n = 3)	94	48	62
F005d1 (n = 3)	102	88	100
F005c3 (L, n = 3)	50	47	62

Example: Pharmacokinetic Study 2 of Intramuscular Administration to Female Dogs

Fulvestrant pharmaceutical compositions L3F and L6 were prepared as described elsewhere herein and in the Figures. A preclinical study was performed to determine the pharmacokinetics of the pharmaceutical compositions following a single intramuscular administration of 15.4 mg/kg to female dogs. The pharmacokinetics of 15.4 mg/kg IM FASLODEX™ (fulvestrant injection, 250 mg/5 mL) were also determined and used for comparison to the three prototype pharmaceutical compositions. The 15.4 mg/kg dose used in this study is the canine equivalent, in mg/m², of the maximum dose (500 mg) for human use and was scaled for use in canine by dividing the dose (based on a 60 kg human) by a canine species conversion factor of 0.54.
Nine non-naïve female beagle dogs were used in the study. The animals weighed between approximately 5-13 kg. Animal welfare for this study was in compliance with the U.S. Department of Agriculture's (USDA) Animal Welfare Act (9 Code of Federal Regulations (CFR) Parts 1, 2 and 3). The Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Academy Press, Washington, D.C., was followed. The facility maintained an Animal Welfare Assurance statement with the National Institutes of Health, Office of Laboratory Animal Welfare.
The FASLODEX™ test articles contained a small molecule that was used as received and no adjustment was made for purity, salt correction, etc. The FASLODEX™ test articles were gently agitated prior to dispensing and dose delivery. Pharmaceutical Compositions L3F and L6 were stored at room temperature and protected from light prior to use, and gently agitated prior to dispensing and dose delivery.

TABLE 38

			Fulvestrant
		Dosage	Conc.	Dose Volume
Dose Group	Test Article	(mg/kg)	(mg/mL)	(mg/kg)

1	FASLODEX ™	15.4	50	0.308
2	Pharmaceutical	15.4	100	0.154
	Composition
	L3F

3	Pharmaceutical	15.4	100	0.154
	Composition
	L6

The animals were not fasted prior to dosing. Each animal received a single intramuscular (IM) dose of only one of the appropriate test article pharmaceutical compositions as outlined in the following study design table, Table 38. IM doses were administered with a 20 G needle via bolus injection into the same large muscle mass (using the Z-track injection technique) in the left hind limb of each animal. Attempts were made for consistent injections between animals [selection of the dose site (muscle), depth, etc.]. The hair was clipped from the injection site prior to dosing. The injection site was marked following dosing and remarked as necessary throughout the study. Specifications for all dose delivery were recorded and reported in the study report [including, but not limited to needle gauge/length, syringe size/barrel type with manufacturer and part number, estimated injection depth into the muscle, approximate duration required to administer the injection; any substantial resistance (either flow through the syringe/needle and/or into the muscle during administration)] was documented.
All animals were observed at least twice a day for morbidity, mortality, injury, and availability of food and water. Any animals in poor health were identified for further monitoring and possible euthanasia.
Whole venous blood samples of approximately 2 mL each were collected from a peripheral vein of all animals for determination of fulvestrant exposure. Samples were collected at the following target timepoints; predose, 0.25, 0.5, 1, 2, 4, 8, 12, 24 (Day 2), 48 (Day 3), 72 (Day 4), 96 (Day 5), 120 (Day 6), 192 (Day 9), 264 (Day 12), 336 (Day 15), 384 (Day 17), 528 (Day 23), and 672 (Day 29) hours after administration. Blood was collected with sodium heparin anticoagulant (glass tube, no gel separator). All blood samples were placed on wet ice following collection until centrifuged. Blood was centrifuged at 3500 rpm for 7 minutes at 2 to 8° C. Plasma (minimum of 0.8 mL volume) was separated from blood cells within 0.75 hours of blood collection and frozen. Plasma samples were initially placed on dry ice prior to being stored in the appropriate freezer (−60 to −90° C.). Samples were shipped on dry ice for bioanalytical analysis.
A model independent method was used to determine C_maxand AUC values from fulvestrant plasma concentration-time data. Results are shown in Tables 31-37 and FIGS. 2A and 2C. Table 31 shows the pharmacokinetic data from the 15.4 mg/kg dosages as nominally dosed (based on the target fulvestrant concentration for each pharmaceutical composition). An “Assay %” is shown for the fulvestrant pharmaceutical compositions of the present disclosure used in the study. The “Assay %” represents the percentage equivalence of the particular pharmaceutical composition in comparison to the FASLODEX™ label claim fulvestrant concentration, with “Assay %” values determined via HPLC, measurement samples taken pre- and post-dose, with one value selected for normalization. The data in Tables 32-37 are normalized using the “Assay %” values to compare PK results based upon the actual mg/kg of fulvestrant administered, assuming linear scaling. FIGS. 2A and 2C depict graphs of the dose normalized fulvestrant mean plasma concentrations.

Example: Pharmacokinetic Study 3 of Intramuscular and Intravenous Administration to Female Dogs

Fulvestrant formulations F003a, F003b, F004a, F003e, F003k2, F003k3, F005a2, F0031, F005b1, F015a1, F005d1, F005c3, F015a3, F005g5, Del-1S, Del-2S, F005H3, Lot 15, Lot 26, Lot 27, Lot 28, Lot 42, Lot 43, Lot 45, Lot 46, Lot 47, and Lot 48 were prepared as described elsewhere herein and in the Figures. In some instances, the formulations were prepared using different processes as indicated, referred to by an alphanumeric process identifier, such as “Process A1,” “Process A2,” and the like. A fulvestrant pharmaceutical composition for intravenous injection, referred to as batch FV-004/15M, was prepared as described below. A preclinical study was performed to determine the pharmacokinetics of the pharmaceutical compositions following a single intramuscular (IM) administration of 15.4 mg/kg to female dogs. The pharmacokinetics of 15.4 mg/kg IM FASLODEX™ (fulvestrant injection, 250 mg/5 mL) were also determined and used for comparison to the three prototype pharmaceutical compositions. The 15.4 mg/kg dose used in this study is the canine equivalent, in mg/m², of the maximum dose (500 mg) for human use and was scaled for use in canine by dividing the dose (based on a 60 kg human) by a canine species conversion factor of 0.54.
One hundred fifty-six non-naïve female beagle dogs of body weight range of 5.65 to 11.40 kilograms were used in the study and assigned to Groups 1-48, as shown in Table 39 below.
Animal welfare for this study was in compliance with the U.S. Department of Agriculture's (USDA) Animal Welfare Act (9 Code of Federal Regulations (CFR) Parts 1, 2 and 3). The Guide for the Care and Use of Laboratory Animals, Institute of Laboratory Animal Resources, National Academy Press, Washington, D.C., was followed. The facility maintained an Animal Welfare Assurance statement with the National Institutes of Health, Office of Laboratory Animal Welfare.

TABLE 39

				Fulvestrant	Dose	Dose
Group		Number of	Dose	Concentration	Level	Volume
Number	Test Article	Females	Route	(mg/mL)	(mg/kg)	(mL/kg)

1	Faslodex (lot	3	IM	50	15.4	0.308
	LW466)
2	Faslodex (lot	3	IM	50	15.4	0.308
	MB122)
3	Faslodex (lot	3	IM	50	15.4	0.308
	MB948)
4	Formulation F003a	3	IM	100	15.4	0.154
5	Formulation F003b	3	IM	100	15.4	0.154
6	Formulation F004a	3	IM	100	15.4	0.154
7	Formulation F003e	3	IM	100	15.4	0.154
8	Formulation F003k2	4	IM	100	15.4	0.154
9	Formulation F003k3	4	IM	100	15.4	0.154
10	Faslodex (lot	4	IM	50	15.4	0.308
	MC949)
11	Formulation F005a2	3	IM	100	15.4	0.154
12	Formulation F003l	4	IM	100	15.4	0.154
13	Formulation F005b1	3	IM	100	15.4	0.154
14	Formulation F015a1	3	IM	100	15.4	0.154
15	Formulation F005d1	3	IM	100	15.4	0.154
16	Formulation F005c3	4	IM	100	15.4	0.154
17	Formulation F015a3	3	IM	100	15.4	0.154
18	Fulvestrant	4	IV	20	2.5	0.125
	(batch FV-004/15M)
19	Formulation F005g5	4	IM	100	15.4	0.154
20	Formulation Del-1S	4	IM	100	15.4	0.154
21	Formulation Del-2S	4	IM	100	15.4	0.154
22	Formulation F005H3	3	IM	100	15.4	0.154
23	Lot 15, by Process	4	IM	100	15.4	0.154
	E1
24	Lot 15, by Process	3	IM	100	15.4	0.154
	E2
25	Lot 26, by Process F1	3	IM	100	15.4	0.154
26	Lot 26, by Process F2	3	IM	100	15.4	0.154
27	Lot 26, by Process F3	3	IM	100	15.4	0.154
28	Lot 26, by Process F4	3	IM	100	15.4	0.154
29	Lot 26, by Process J1	3	IM	100	15.4	0.154
30	Lot 26, by Process J2	3	IM	100	15.4	0.154
31	Lot 26, by Process J3	3	IM	100	15.4	0.154
32	Lot 26, by Process J4	3	IM	100	15.4	0.154
33	Lot 42, by Process G1	3	IM	100	15.4	0.154
34	Lot 42, by Process G2	3	IM	100	15.4	0.154
35	Lot 43, by Process H1	3	IM	100	15.4	0.154
36	Lot 43, by Process H2	3	IM	100	15.4	0.154
37	Lot 27, by Process A1	3	IM	100	15.4	0.154
38	Lot 27, by Process A2	3	IM	100	15.4	0.154
39	Lot 27, by Process A3	3	IM	100	15.4	0.154
40	Lot 27, by Process A4	3	IM	100	15.4	0.154
41	Lot 28, by Process B1	3	IM	100	15.4	0.154
42	Lot 28, by Process B2	3	IM	100	15.4	0.154
43	Lot 28, by Process B3	3	IM	100	15.4	0.154
44	Lot 28, by Process B4	3	IM	100	15.4	0.154
45	Lot 45, by Process C1	3	IM	100	15.4	0.154
46	Lot 46	3	IM	100	15.4	0.154
47	Lot 47	3	IM	100	15.4	0.154
48	Lot 48	3	IM	100	15.4	0.154

Pharmaceutical Compositions F003a, F003b, F004a, F003e, F003k2, F003k3, F005a2, F0031, F005b1, F015a1, F005d1, F005c3, F015a3, FV-004/15M, Del-1S, and Del-2S were stored at room temperature and protected from light prior to use, and gently agitated via inversion prior to dispensing and dose delivery. If visible clumps of material were seen on the vial inside wall or inner seal of the vial cap after 3 minutes of inversion, the tightly capped vial was vortexed at moderate intensity and unlimited duration until clumps were not visible. FASLODEX™ was supplied as two 5-mL clear neutral glass (Type 1) syringe barrels, each containing a 250 mg/5 mL (50 mg/mL) solution for intramuscular injection. Upon receipt, FASLODEX™ was stored refrigerated (2°-8° C.) and protected from light. The procedure to prepare and administer FASLODEX™ was performed as outlined in the manufacturer's prescribing information.
Lyophilized pharmaceutical compositions of formulations F005g5, F005H3, F015a3, Lot 15, Lot 26, Lot 27, Lot 28, Lot 42, Lot 43, Lot 45, Lot 46, Lot 47, and Lot 48 were reconstituted prior to dosing. Using an empty syringe and hypodermic needle, about 5 mL of air was withdrawn from the head space of the vial (above the lyophilized contents) via the septum and the syringe and needle were discarded. Using a sterile syringe and hypodermic needle, 5 mL of sterile water for injection, USP were added to the vial by piercing the septum and injecting a stream of water slowly around the inner wall of the neck of the vial to wet the lyophilized cake without touching any of the vial contents. The needle was removed from the septum and the vial was gently swirled until a visually homogeneous particulate suspension formed, with no visual clumps or material attached to the inside wall of the vial. If a homogeneous suspension was not formed after 5 minutes of swirling, the vial was vortexed until a homogeneous suspension was formed. The vials were not shaken to avoid generating bubbles or excessive foam.
Group 18 was administered an intravenous batch of fulvestrant (batch FV-004/15M) prepared as follows by (% w/v): 2% fulvestrant, 10% EtOH, 79% propylene glycol, 1% Poloxamer 407, 8% Water for Injection, USP. Fulvestrant API was stored at 2-8° C., protected from light. Care was taken to protect the API from humidity during weighing. Fulvestrant powder was dissolved in ethanol and swirled and vortexed as needed to dissolve completely. Propylene glycol was added and mixed to dissolve to a clear liquid state. Poloxamer 407 was dissolved in water for injection, USP in a separate vessel and mixed, vortexed, and sonicated as needed to dissolve into to a clear liquid state. The Poloxamer 407 in water for injection solution was added to the fulvestrant/ethanol solution. Propylene glycol was added and the solution was mixed and vortexed to achieve a clear liquid. The solution was filtered through a 0.2 μm or 0.22 μm syringe (to ensure that all liquid volume was usable) tip filter (PVDF) into a clear glass vessel. The prepared formulation as stored at room temperature for up to four hours prior to dosing under protection from light exposure. Intravenous doses were administered via the cephalic (or other suitable) vein as a slow injection over approximately 1 minute. Batch FV-004/15M was administered intravenously at a dose of 2.5 mg/kg.
The animals were not fasted prior to dosing. Each animal in Groups 4-9, 11-17, and 19-48 received a single intramuscular (IM) dose of only one of the appropriate test article pharmaceutical compositions as outlined in Table 39. IM doses were administered with a 20 G needle via bolus injection into the same large muscle mass (using the Z-track injection technique) in the left hind limb of each animal. Attempts were made for consistent injections between animals [selection of the dose site (muscle), depth, etc.]. The hair was clipped from the injection site prior to dosing. The injection site was marked following dosing and remarked as necessary throughout the study. Specifications for all dose delivery were recorded and reported in the study report [including, but not limited to needle gauge/length, syringe size/barrel type with manufacturer and part number, estimated injection depth into the muscle, approximate duration required to administer the injection; any substantial resistance (either flow through the syringe/needle and/or into the muscle during administration)] was documented. Animals in Groups 13-17 and 19-48 were administered 1 tablet or capsule (25 mg) of (PO) diphenhydramine at approximately 1 hour prior to dosing.
All animals were observed at least twice a day for morbidity, mortality, injury, and availability of food and water. Any animals in poor health were identified for further monitoring and possible euthanasia.
Whole venous blood samples of approximately 2 mL each were collected from a peripheral vein of all animals for determination of fulvestrant exposure. Blood samples for Groups 1-7 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, and 12 (on Day 1); and 24 (on Day 2), 48 (on Day 3), 72 (on Day 4), 96 (on Day 5), 120 (on Day 6), 192 (on Day 9), 264 (on Day 12), 336 (on Day 15), 384 (on Day 17), 528 (on Day 23), and 672 (on Day 29) hours postdose. Blood samples for Groups 8-14 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, and 12 (on Day 1); and 24 (on Day 2), 48 (on Day 3), 72 (on Day 4), 96 (on Day 5), 120 (on Day 6), 192 (on Day 9), 264 (on Day 12), 336 (on Day 15), 384 (on Day 17), 456 (on Day 20), 528 (on Day 23), 600 (on Day 26), 672 (on Day 29), 696 (Day 30), 768 (Day 33), 816 (Day 35), 864 (Day 37), 936 (Day 40) and 1008 (Day 43) hours post-dose. Blood samples for Groups 15-17 and 19-21 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, 12 (Day 1), 24 (Day 2), 48 (Day 3), 72 (Day 4), 96 (Day 5), 120 (Day 6), 192 (Day 9), 264 (Day 12), 336 (Day 15), 384 (Day 17), 456 (Day 20), 528 (Day 23), 600 (Day 26), 672 (Day 29), 696 (Day 30), 768 (Day 33), 816 (Day 35), 864 (Day 37), 936 (Day 40), 1008 (Day 43), and 1176 (Day 50) hours postdose. Blood samples for Group 18 were collected predose and at 0.033 (2 minutes), 0.1 (6 minutes), 0.13 (8 minutes), 0.27 (16 minutes), 0.52 (31 minutes), 0.77 (46 minutes), 1, 2, 3, 4, 6, 8, 10, and 12 hours postdose on Day 1 and at 24 (on Day 2), 30 (on Day 2), 48 (on Day 3) and 72 (on Day 4) hours postdose, with postdose measurements from the start of dose administration, which took about 1 minute to complete. Blood samples for Groups 22-48 were collected predose and at 0.25, 0.5, 1, 2, 4, 8, 12 (Day 1), 24 (Day 2), 48 (Day 3), 72 (Day 4), 96 (Day 5), 120 (Day 6), 144 (Day 7), 168 (Day 8), 192 (Day 9), 216 (Day 10), 240 (Day 11), 264 (Day 12), 336 (Day 15), 384 (Day 17), 456 (Day 20), 528 (Day 23), 600 (Day 26), 672 (Day 29), 696 (Day 30), 768 (Day 33), 816 (Day 35), 864 (Day 37), 936 (Day 40), 1008 (Day 43), and 1176 (Day 50) hours postdose. Blood was collected with sodium heparin anticoagulant (glass tube, no gel separator). All blood samples were placed on wet ice following collection until centrifuged. Blood was centrifuged at 3500 rpm for 7 minutes at 2 to 8° C. Plasma (minimum of 0.8 mL volume) was separated from blood cells within 0.75 hours of blood collection and frozen. Plasma samples were initially placed on dry ice prior to being stored in the appropriate freezer (−60 to −90° C.). Samples were shipped on dry ice for bioanalytical analysis to determine absolute ng/mL fulvestrant in the plasma.

	TABLE 40

	PK parameters compared to
	Faslodex lots based upon the
	nominal dose 15.4 mg/kg

			AUC_{0-14 d}	AUC_{0-28 d}
Formulation	PK parameters based upon the	Cmax	ratio	ratio
(Geometric Mean,	nominal dose 15.4 mg/kg	ratio to	to	to

n = 3 unless indicated	Cmax	AUC_{0-14 d}	AUC_{0-28 d}	Faslodex	Faslodex	Faslodex
otherwise)	(ng/mL)	(hr * ng/mL)	(hr * ng/mL)	(%)	(%)	(%)

Faslodex (All, n = 22)	37.5	7408	9195	100	100	100
F005g5	32.1	6950	10400	86	94	113
Lot 15, by Process E1	20.1	3890	5500	54	53	60
Lot 15, by Process E2	20.3	4190	6230	54	57	68
Lot 26, by Process F1	19.5	4150	6650	52	56	72
Lot 26, by Process F2	30.5	5510	7850	81	74	85
Lot 26, by Process F3	25.7	5350	8260	68	72	90
Lot 26, by Process F4	30.8	7210	10200	82	97	111
Lot 26, by Process J1	25.2	5490	7910	67	74	86
Lot 26, by Process J2	20.8	4640	7190	55	63	78
Lot 26, by Process J3	21.1	4720	7310	56	64	80
Lot 26, by Process J4	21.3	4320	5900	57	58	64
Lot 42, by Process G1	28.1	4640	7410	75	63	81
Lot 42, by Process G2	33.8	6110	8800	90	82	96
Lot 43, by Process H1	25.0	5190	7500	67	70	82
Lot 43, by Process H2	22.2	4430	6250	59	60	68
Lot 27, by Process A1	92.6	6150	8710	247	83	95
Lot 27, by Process A2	27.9	6240	9170	74	84	100
Lot 27, by Process A3	26.3	5100	7680	70	69	84
Lot 27, by Process A4	29.1	6030	8620	78	81	94
Lot 28, by Process B1	21.9	3950	6230	58	53	68
Lot 28, by Process B2	36.3	7010	9870	97	95	107
Lot 28, by Process B3	31.6	6880	9660	84	93	105
Lot 28, by Process B4	31.0	6330	9000	83	85	98
Lot 45, by Process C1	29.8	5320	8560	79	72	93
Lot 46	30.2	5570	8240	80	75	90
Lot 47	27.9	5230	8070	74	71	88
Lot 48	26.3	4900	7410	70	66	81

A model independent method was used to determine C_maxand AUC values from fulvestrant plasma concentration-time data. Results are shown in Tables 31-37 and 40 and FIGS. 1A, 1B, 2A, 2B, 2C, 13, and 14. Table 31 shows the pharmacokinetic data from the 15.4 mg/kg dosages as nominally dosed (based on the target fulvestrant concentration for each pharmaceutical composition). An “Assay %” is shown in Table 31 for the fulvestrant pharmaceutical compositions of the present disclosure used in the study. The “Assay %” represents the percentage equivalence of the particular pharmaceutical composition in comparison to the FASLODEX™ label claim fulvestrant concentration, with “Assay %” values determined via HPLC, measurement samples taken pre- and post-dose, with one value selected for normalization. The data in Tables 32-37 are normalized using the “Assay %” values to compare PK results based upon the actual mg/kg of fulvestrant administered, assuming linear scaling. FIGS. 1A, 1B, 2A, 2B, and 2C depict graphs of the dose normalized fulvestrant mean plasma concentrations. Table 40 shows the pharmacokinetic data from the 15.4 mg/kg dosages as nominally dosed (based on the target fulvestrant concentration for each pharmaceutical composition) in comparison to the geometric mean of all Faslodex lots tested (n=22). FIGS. 13 and 14 depict fulvestrant plasma measurements for administration of Faslodex Lot MB948 to three female dogs and administration of fulvestrant formulation Lot 27 processed by Process A2 to three female dogs (referred to in FIG. 14 as subjects 924, 925, and 926).

Example: Microscopic Imaging of Fulvestrant Particles in Suspensions

Some exemplary fulvestrant pharmaceutical compositions of the present disclosure were examined via optical and scanning electron microscopy. Suspensions of fulvestrant pharmaceutical compositions Variants B, E, I, J, K, L, L3F, L6, F003a, F003b, F004a, F003e, F00k2, and F003k3 were examined via optical microscopy. Optical microscopy was performed at 400× magnification with a polarized light filter using fully dispersed homogeneous suspension samples.

Example: Particle Size Distribution Characterization of Fulvestrant Pharmaceutical Compositions

Batches of fulvestrant pharmaceutical composition Lot 27, described elsewhere herein, were prepared by the methods of preparation 101 of FIG. 15. Samples were taken periodically during high shear mixing, prior to any high pressure homogenization steps. Some test samples, referred to as “Sample 1”, were taken after approximately five hours of high shear mixing and other test samples, referred to as “Sample 2”, were taken after approximately 13.7 hours to high shear mixing. Particle sizes of test samples were analyzed using optical microscopy with a Malvern Morphologi G3 apparatus for microscopy image capture and analysis. CE diameters were measured and number-weighted and volume-weighted particle size distribution parameters were determined as shown in Tables 41 and 42. The CE diameter ranges of measurement aliquots are also shown, with the lower range value of 0.54 microns representing the lower limit of detection for the apparatus setup. Test samples were analyzed with a Malvern Mastersizer 3000 apparatus for laser diffraction particle size characterization of LD diameters.

TABLE 41

Volume-Weighted Distribution Parameters

				Circle
	CE	CE		Equivalent
	Dv	Dv	CE	(CE)-	LD	LD	LD
	(10)	(50)	Dv (90)	diameter-	Dv10	Dv50	Dv90
Formulation	(μm)	(μm)	(μm)	range (μm)	(μm)	(μm)	(μm)

Lot 27	6.113	13.77	32.71	0.54-49.72	1.81	6.68	16.6
(Sample 1)
Lot 27	6.509	14.34	28.64	0.54-54.14	1.85	6.93	17.7
(Sample 1)
Lot 27	6.378	12.76	23.90	0.54-55.04
(Sample 1)
Lot 27	5.297	10.73	24.57
(Sample 1)
Lot 27	6.015	13.03	25.25
(Sample 1)
Lot 27	5.446	11.41	22.10
(Sample 1)
Lot 27	7.222	14.93	28.99
(Sample 1)
Lot 27	8.747	18.31	32.93
(Sample 1)
Lot 27	7.663	14.96	26.58
(Sample 1)
Lot 27	6.733	13.61	24.70	0.54-42.64	1.96	7.54	19.3
(Sample 2)
Lot 27	7.182	14.12	25.15	0.54-44.65
(Sample 2)
Lot 27	6.920	13.26	22.63	0.54-38.24
(Sample 2)
Lot 27	7.400	14.86	27.82
(Sample 2)
Lot 27	6.857	14.08	27.39
(Sample 2)
Lot 27	8.117	16.58	30.90
(Sample 2)

TABLE 42

Number-Weighted Distribution Parameters

				Circle
				Equivalent
				(CE)-
	CE Dn10	CE Dn50	CE Dn90	diameter-
Formulation	(μm)	(μm)	(μm)	range (μm)

Lot 27 (Sample 1)	1.10	3.92	8.91	0.54-49.72
Lot 27 (Sample 1)	1.14	4.06	9.61	0.54-54.14
Lot 27 (Sample 1)	0.95	4.16	9.85	0.54-55.04
Lot 27 (Sample 2)	1.08	4.24	10.34	0.54-42.64
Lot 27 (Sample 2)	0.85	3.71	10.52	0.54-44.65
Lot 27 (Sample 2)	0.91	4.07	10.55	0.54-38.24

Fulvestrant pharmaceutical compositions F005g5, Lot 27, Lot 28, and Lot 45, described elsewhere herein, were prepared by the methods of preparation shown schematically as process 101 of FIG. 15. Samples of each Lot were prepared using different processes as indicated in Tables 43 to 50 to achieve fulvestrant particle sizes and particle size distributions. The preparation processes are referred to with alphanumeric identifiers, such as “Process A1,” “Process A2,” and the like, with each process representing a set of fulvestrant-particle-size-reduction steps, as more fully described elsewhere herein, to achieve final fulvestrant particle size and particle size distributions as shown in the Tables 43 to 50. Test samples were evaluated for particle size both prior to drying via lyophilization, which are indicated as “(100× Suspensions)”, and after lyophilization and reconstitution, which are indicated as “(100× Reconstituted Suspensions)” in the Tables 43 to 50. Samples were analyzed with a Malvern Morphologi G3 apparatus for microscopy image capture and analysis. CE diameters were measured and volume-weighted particle size distribution parameters were determined. The CE diameter ranges of measurement samples are also shown, with the lower range value of 0.54 microns representing the lower limit of detection for the apparatus setup. Samples were analyzed with a Malvern Mastersizer 3000 apparatus for laser diffraction particle size characterization of LD diameters.

TABLE 43

Volume-Weighted Distribution Parameters
(100x Reconstituted Suspensions)

					Circle
					Equivalent
		CE	CE	CE	(CE)-
		Dv10	Dv50	Dv90	diameter-
Formulation	Process	(μm)	(μm)	(μm)	range (μm)

Lot 27	Process	6.606	16.14	36.36	0.54-59.46
	A1
	Process	7.031	31.23	84.50	0.54-107.10
	A2
	Process	6.725	24.83	53.11	0.54-84.16
	A3
	Process	6.790	43.74	98.32	0.54-105.94
	A4
Lot 28	Process	5.333	17.64	41.18	0.54-53.33
	B1
	Process	4.863	12.71	42.29	0.54-75.19
	B2
	Process	5.376	15.85	49.64	0.54-75.62
	B3
	Process	4.144	25.86	82.19	0.54-107.08
	B4
Lot 45	Process	8.904	35.32	66.41	0.54-85.05
	C1
	Process	8.135	34.79	56.77	0.54-81.50
	C2

TABLE 44

Volume-Weighted Distribution Parameters
(100x Reconstituted Suspensions)

	Circle
	Equivalent	As-Is	Sonicated

		CE	CE	CE	(CE)-	LD	LD	LD	LD	LD	LD
		Dv10	Dv50	Dv90	diameter-	Dv10	Dv50	Dv90	Dv10	Dv50	Dv90
Formulation	Process	(μm)	(μm)	(μm)	range (μm)	(μm)	(μm)	(μm)	(μm)	(μm)	(μm)

Lot 45	Process	5.140	14.44	42.62	0.54-71.31	3.42	8.21	33.8	2.81	5.57	10.6
	C9
Lot 45	Process	6.558	17.10	44.70	0.54-74.57	3.37	7.81	32.6	2.82	5.58	10.5
	C9
Lot 45	Process	6.648	19.97	56.47	0.54-112.57	3.50	9.34	43.1	2.81	5.57	10.5
	C9
Lot 45	Process	6.541	15.68	37.75	0.54-55.93	3.20	7.08	24.3	2.80	5.52	10.7
	C10
Lot 45	Process	5.787	13.11	32.25	0.54-55.82	3.33	7.83	33.9	2.98	5.80	11.1
	C10
Lot 45	Process	5.375	14.59	55.02	0.54-88.17	3.38	8.02	35.6	2.97	5.79	11.1
	C10
F005g5	As					3.07	29.2	95.1	2.20	5.25	14.0
	shown
	in
	Table
	34

TABLE 45

Volume-Weighted Distribution Parameters
(100x Reconstituted Suspensions)

					Circle
					Equivalent
		CE			(CE)-
		Dv10	CE Dv50	CE Dv90	diameter-
Formulation	Process	(μm)	(μm)	(μm)	range (μm)

Lot 27	Process A2	4.547	11.10	33.19	0.54-55.75
Lot 27	Process A2	4.290	10.15	23.66	0.54-49.72
Lot 27	Process A2	5.908	12.98	25.55	0.54-42.63
Lot 27	Process A2	6.075	12.81	23.11	0.54-47.71
Lot 27	Process A2	7.022	15.59	30.94	0.54-69.41
Lot 27	Process A2	5.399	12.22	34.36	0.54-51.56
Lot 27	Process A13	4.89	11.84	32.47
Lot 27	Process A13	6.46	15.33	32.47
Lot 27	Process A13	4.93	11.90	31.23
Lot 27	Process A13	5.54	14.17	50.26
Lot 27	Process A13	4.19	10.16	32.22
Lot 27	Process A13	5.91	13.71	33.81
Lot 27	Process A13	5.10	13.26	45.32
Lot 27	Process A13	4.61	11.47	39.79
Lot 27	Process A13	4.79	12.50	46.19
Lot 27	Process A13	4.71	10.33	22.64
Lot 27	Process A13	5.66	12.66	29.72
Lot 27	Process A13	4.96	11.62	37.81
Lot 27	Process A13	4.22	12.46	42.59
Lot 27	Process A13	5.80	19.69	57.80
Lot 27	Process A13	5.12	11.53	28.37
Lot 27	Process A13	3.84	9.28	32.42
Lot 27	Process A13	4.64	10.32	25.28
Lot 27	Process A13	4.86	11.69	32.44

TABLE 46

Volume-Weighted Distribution Parameters
(100x Reconstituted Suspensions)

As-Is

Sonicated

		LD	LD	LD	LD		LD
		Dv10	Dv50	Dv90	Dv10	LD Dv50	Dv90
Formulation	Process	(μm)	(μm)	(μm)	(μm)	(μm)	(μm)

Lot 27	Process	2.03	8.41	36.0	1.20	3.93	7.97
	A13
Lot 27	Process	1.96	7.83	35.0	1.14	3.69	7.57
	A13
Lot 27	Process	2.08	9.14	41.3	1.20	3.92	7.94
	A13
Lot 27	Process	2.10	9.65	40.3	1.15	3.66	7.53
	A13
Lot 27	Process	1.67	6.10	26.5	1.08	3.42	7.19
	A13
Lot 27	Process	1.70	5.71	21.6	1.17	3.80	7.78
	A13
Lot 27	Process	2.53	15.8	48.4	1.27	4.07	8.72
	A13
Lot 27	Process	2.01	12.6	57.6	1.01	3.04	6.85
	A13
Lot 27	Process	1.62	6.34	29.5	0.96	2.76	5.89
	A13

TABLE 47

Volume-Weighted Distribution Parameters
(100x Suspensions)

Lot 27	Process A15	5.199	10.4	17.52	0.54-27.89
	Process A16	3.766	7.411	12.15	0.54-25.43
	Process A17	3.053	6.256	10.81	0.54-38.27
	Process A18	3.727	7.277	13.91	0.54-31.30
Lot 28	Process B5	4.442	8.585	15.04	0.54-41.23
	Process B6	4.134	7.773	13.25	0.54-22.59
	Process B7	3.835	7.311	13.11	0.54-25.94
	Process B8	3.417	6.644	11.62	0.54-28.25
Lot 45	Process C3	7.160	14.79	25.70	0.54-47.55
	Process C4	8.587	18.15	36.68	0.54-62.10

TABLE 48

Volume-Weighted Distribution Parameters
(100x Suspensions)

As-Is

Sonicated

		LD	LD		LD	LD	LD
		Dv10	Dv50	LD Dv90	Dv10	Dv50	Dv90
Formulation	Process	(μm)	(μm)	(μm)	(μm)	(μm)	(μm)

Lot 27	Process	1.08	3.29	7.68	0.91	2.62	6.98
	A15
	Process	1.03	3.15	7.00	0.83	2.32	5.72
	A16
	Process	0.97	2.96	6.52	0.78	2.13	5.01
	A17
	Process	0.92	2.91	6.32	0.72	1.96	4.38
	A18
Lot 45	Process	1.50	6.03	12.0	0.83	2.30	5.69
	C3
	Process	1.59	5.82	11.0	0.85	2.37	5.74
	C4

TABLE 49

Volume-Weighted Distribution Parameters
(100x Suspensions)

	Circle
	Equivalent	As-Is	Sonicated

Lot 45	Process	12.320	29.88	57.24	0.54-90.58	1.51	5.40	10.7	0.80	2.11	4.64
	C3
Lot 45	Process	7.454	17.37	40.29	0.54-76.61	1.71	6.36	12.5	0.82	2.26	5.20
	C4
Lot 45	Process	12.130	27.59	50.08	0.54-87.69	1.73	6.38	12.7	0.79	2.15	4.77
	C5
Lot 45	Process	8.193	17.89	35.05	0.54-54.50	1.63	5.85	11.7	0.79	2.13	4.68
	C5
Lot 45	Process	8.690	19.36	35.53	0.54-64.77	1.70	6.12	12.0	0.77	2.05	4.45
	C5
Lot 45	Process	8.218	17.00	46.80	0.54-80.21	1.46	4.92	9.59	1.36	4.71	9.55
	C6
Lot 45	Process	6.455	14.94	38.47	0.54-65.55	1.27	4.21	8.61	0.77	2.11	4.66
	C7
Lot 45	Process	7.795	17.20	31.60	0.54-45.03	1.55	5.37	10.7	0.77	2.14	4.81
	C8
Lot 45	Process	9.257	20.16	39.47	0.54-84.26	1.64	5.64	11.2	0.78	2.19	4.99
	C8
Lot 45	Process	10.050	21.35	36.43	0.54-59.43	1.59	5.55	11.1	0.76	2.10	4.66
	C8

TABLE 50

Volume-Weighted Distribution Parameters
(100x Suspensions)

					Circle
					Equivalent	As-Is	As-Is	As-Is
		CE	CE	CE	(CE)-	LD	LD	LD
		Dv10	Dv50	Dv90	diameter-	Dv10	Dv50	Dv90
Formulation	Process	(μm)	(μm)	(μm)	range (μm)	(μm)	(μm)	(μm)

Lot 27	Process A5	5.506	12.94	26.24	0.54-52.71	1.54	5.70	11.3
Lot 27	Process A6	8.740	19.30	35.62	0.54-54.43	1.42	5.10	10.1
Lot 27	Process A7	7.862	18.92	36.09	0.54-52.63	1.48	5.30	10.4
Lot 27	Process A8	8.184	17.49	33.02	0.54-63.56	1.46	5.04	9.69
Lot 27	Process A9	7.451	15.99	29.94	0.54-55.58	1.49	5.00	9.65
Lot 27	Process A10	7.673	18.50	36.71	0.54-74.22	1.59	5.55	10.6
Lot 27	Process A11	9.093	20.01	45.26	0.54-61.26	1.72	6.11	12.1
Lot 27	Process A12	6.355	14.92	46.31	0.54-63.98	1.66	5.77	10.9
Lot 27	Process A13	7.029	15.66	31.06	0.54-55.89	1.07	3.51	7.32
Lot 27	Process A14	7.870	20.15	43.79	0.54-62.35	1.74	5.87	11.0
Lot 27	Process A14	8.499	19.06	37.90	0.54-70.88	1.67	5.49	10.5
Lot 27	Process A14	8.072	17.43	30.90	0.54-56.50	1.76	6.01	11.5
Lot 27	Process A14	8.078	18.98	45.32	0.54-69.82	1.81	6.43	12.5

When ranges are used herein for chemical or physical properties, such as particle size or particle size distribution, formulation component concentrations, or pharmacokinetic properties, all combinations, and subcombinations of ranges for specific embodiments therein are intended to be included.
The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in its entirety.
Those skilled in the art will appreciate that numerous changes and modifications can be made to the preferred embodiments of the invention and that such changes and modifications can be made without departing from the spirit of the invention. It is, therefore, intended that the appended claims cover all such equivalent variations as fall within the true spirit and scope of the invention.

Claims

1. A suspension comprising fulvestrant particles and a vehicle;

wherein optionally the fulvestrant particles have

an LD Dv(10) between about 1 micron to about 3 microns, preferably an LD Dv(10) of about 1-2 microns,

an LD Dv(50) between about 2 microns and about 35 microns, preferably an LD Dv(50) of about 2-4 microns, and

an LD Dv(90) between about 4 microns and about 120 microns, preferably an LD Dv(90) of about 6-9 microns, and;

wherein optionally the vehicle is a non-oil vehicle, preferably water or non-aqueous vehicle;

wherein optionally fulvestrant is at a concentration of about 100 mg/mL; and

wherein optionally the suspension is substantially oil-free.

2. A suspension comprising fulvestrant and a vehicle,

wherein the fulvestrant is at a concentration equal to or greater than about 50 mg/mL, preferably about 50 mg/mL or about 100 mg/mL;

wherein optionally the suspension is substantially oil-free.

3. A pharmaceutical composition comprising fulvestrant particles and a non-oil vehicle,

wherein optionally the pharmaceutical composition further comprises at least one stabilizer selected from the group consisting of surfactants, polymers, cross-linked polymers, buffering agents, electrolytes, and non-electrolytes, preferably wherein the at least one stabilizer is a cross-linked polymer, preferably carboxymethylcellulose sodium, or the at least one stabilizer is selected from:

the group of polyethylene oxide (PEO), a PEO derivative, polysorbate 80, polysorbate 20, poloxamer 188, poloxamer 124, poloxamer 407, polyethoxylated vegetable oils, polyethoxylated castor oil, sorbitan palmitate, lecithin, poly(vinyl alcohol), human serum albumin, and mixtures thereof,

the group consisting of polyvinylpyrrolidone, povidone K12, povidone K17, PLASDONE™ C-12 povidone, PLASDONE™ C-17 povidone, PLASDONE™ C-30 povidone, polyethylene glycol 3350, and mixtures thereof,

the group consisting of sodium chloride, calcium chloride, and mixtures thereof, or

the group consisting of dextrose, glycerol, mannitol, and mixtures thereof;

wherein optionally the pharmaceutical composition further comprises at least one buffering agent selected from the group consisting of NaH₂PO₄.H₂O, NaH₂PO₄.2H₂O, anhydrous NaH₂PO₄, sodium citrate, citric acid, Tris, sodium hydroxide, HCl, or a mixture thereof; and

wherein optionally the pharmaceutical composition further comprises solubilized fulvestrant.

4. A pharmaceutical composition comprising fulvestrant particles,

wherein the fulvestrant particles have one or more of:

an LD Dv(10) between about 1 micron to about 3 microns, preferably between about 1 micron to about 2 microns or between about 2 microns and about 3 microns;

an LD Dv(50) between about 2 microns and about 35 microns, preferably between about 2 microns to about 6 microns, more preferably between about 2 microns and about 4 microns; and

an LD Dv(90) between about 4 microns and about 120 microns, preferably between about 7 microns to about 15 microns, more preferably between about 12 microns to about 14 microns or between about 9 microns to about 11 microns, or preferably between about 6 microns and about 9 microns, more preferably between about 6 microns and about 8 microns, most preferably between about 7 microns and about 8 microns;

wherein optionally the pharmaceutical composition is in the form of a suspension, preferably an aqueous suspension.

5. A pharmaceutical composition comprising fulvestrant,

wherein the fulvestrant is at a concentration of about 100 mg/mL and wherein upon administration to a subject, the 90% confidence intervals (CI) of the relative mean AUC_(0-t)and AUC_(0-∞)of fulvestrant of the pharmaceutical composition is within 80% to 125% of the relative mean AUC_(0-t)and AUC_(0-∞), respectively, of fulvestrant upon administration of a reference listed fulvestrant product, for example, FASLODEX™;

wherein optionally the reference listed fulvestrant product comprises fulvestrant at a concentration of about 50 mg/mL;

wherein optionally upon administration to a subject, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 40% to 80% of the relative mean Cmax of fulvestrant after administration of the reference listed fulvestrant product.

6. A pharmaceutical composition comprising fulvestrant wherein the fulvestrant is at a concentration of about 100 mg/mL and wherein after an initial administration to a subject of the pharmaceutical composition comprising 500 mg fulvestrant on day 1 and an additional administration of the pharmaceutical composition comprising 500 mg fulvestrant on day 15 to the subject provides:

an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

wherein optionally the pharmaceutical composition is administered as a single injection of about 5 mL or administered as two injections of about 2.5 mL.

7. A pharmaceutical composition comprising fulvestrant particles wherein the fulvestrant is at a concentration of about 40 to about 125 mg/mL.

8. The pharmaceutical composition of claim 7,

wherein at least about 70% of the fulvestrant is present as fulvestrant particles, preferably at least about 80% of the fulvestrant is present as fulvestrant particles, more preferably at least about 90% of the fulvestrant is present as fulvestrant particles.

9. The pharmaceutical composition of claim 7,

wherein no more than about 20% of the fulvestrant is solubilized, preferably no more than about 10% of the fulvestrant is solubilized, more preferably no more than about 5% of the fulvestrant is solubilized.

10. The pharmaceutical composition of claim 6,

wherein the fulvestrant particles:

have one or more of:

an LD Dv(10) between about 1 micron to about 3 microns;

an LD Dv(50) between about 2 microns and about 35 microns; and

an LD Dv(90) between about 4 microns and about 120 microns;

have one or more of:

an LD Dv(10) between about 1-2 microns;

an LD Dv(50) between about 2-4 microns; and

an LD Dv(90) between about 6-9 microns;

have an LD Dv(90) of about 12-14 microns;

have an LD Dv(90) of about 9-11 microns;

have an LD Dv(90) of about 6-9 microns;

have an LD Dv(90) of about 6-8 microns; or

have an LD Dv(90) of about 7-8 microns.

11. A method of forming an aqueous fulvestrant suspension comprising:

mixing an aqueous medium and at least one stabilizer to form a suspension vehicle, the at least one stabilizer preferably comprising one surfactant and one polymer or one surfactant and one non-electrolyte;

adding an amount of fulvestrant to the suspension vehicle; and

dispersing the fulvestrant in the suspension vehicle to form the aqueous fulvestrant suspension, preferably performed using high shear mixing;

optionally further comprising homogenizing the aqueous fulvestrant suspension, the homogenizing preferably performed using high pressure homogenization, preferably at a pressure of about 15,000 psi to about 45,000 psi, the method preferably further comprising adding an electrolyte to the homogenized aqueous fulvestrant suspension and mixing the electrolyte into the suspension or adding a non-electrolyte to the homogenized aqueous fulvestrant suspension and mixing the non-electrolyte into the suspension;

the method optionally further comprising concentrating the fulvestrant suspension by phase separating the suspension and removing a portion of the supernatant;

the method optionally further comprising drying the aqueous suspension to form a dried pharmaceutical composition, the method preferably further comprising sterilizing the dried pharmaceutical composition using gamma irradiation, the method more preferably further comprising reconstituting the dried pharmaceutical composition into a second aqueous suspension by adding at least one of water for injection (WFI), normal saline (NS), and 5% dextrose in water (D5W).

12. An aqueous fulvestrant suspension prepared according to the method of claim 11.

13. A pharmaceutical composition comprising the aqueous fulvestrant suspension of claim 12,

wherein optionally the pharmaceutical composition comprises fulvestrant particles:

having one or more of:

an LD Dv(10) between about 1 micron to about 3 microns;

an LD Dv(50) between about 2 microns and about 35 microns; and

an LD Dv(90) between about 4 microns and about 120 microns;

having one or more of:

an LD Dv(10) between about 1-2 microns;

an LD Dv(90) between about 6-9 microns; and

an LD Dv(50) between about 2-4 microns;

having one or more of:

a CE Dv(90) between about 10 microns and about 200 microns;

a CE Dv(50) between about 5 microns and about 60 microns; and

a CE Dv(10) between about 1 microns and about 25 microns;

having a CE Dv(90) between about 10 microns and about 200 microns;

having a CE Dv(50) between about 5 microns and about 60 microns; or

having a CE Dv(10) between about 1 microns and about 25 microns.

14. An aqueous fulvestrant suspension comprising:

an amount of fulvestrant, preferably about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, or about 250 mg;

about 0.2 mg/mL to about 75 mg/mL of one or more stabilizers; and

an amount of aqueous medium;

wherein upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean AUC(0-t) and AUC(0-∞) of fulvestrant is within 80% to 125% of the relative mean AUC(0-t) and AUC(0-∞), respectively, of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

wherein optionally the suspension has a volume of about 3.0 to about 5.0 mL, preferably about 3.5 to about 4.5 mL, more preferably about 4.0 mL.

15. The aqueous fulvestrant suspension of claim 14,

wherein the one or more stabilizers:

is selected from the group consisting of surfactants, polymers, cross-linked polymers, buffering agents, electrolytes, and non-electrolytes,

is selected from the group consisting of polyethylene oxide (PEO), a PEO derivative, polysorbate 80, polysorbate 20, poloxamer 188, poloxamer 124, poloxamer 407, polyethoxylated vegetable oils, polyethoxylated castor oil, sorbitan palmitate, lecithin, poly(vinyl alcohol), human serum albumin, and mixtures thereof,

is selected from the group consisting of polyvinylpyrrolidone, povidone K12, povidone K17, PLASDONE™ C-12 povidone, PLASDONE™ C-17 povidone, PLASDONE™ C-30 povidone, polyethylene glycol 3350, and mixtures thereof,

is selected from the group consisting of sodium chloride, calcium chloride, and mixtures thereof, or

is selected from the group consisting of dextrose, glycerol, mannitol, and mixtures thereof.

16. The aqueous fulvestrant suspension of claim 14, wherein the one or more stabilizers comprises a cross-linked polymer, preferably carboxymethylcellulose sodium.

17. The aqueous fulvestrant suspension of claim 14, further comprising at least one buffering agent selected from the group consisting of NaH₂PO₄.H₂O, NaH₂PO₄.2H₂O, anhydrous NaH₂PO₄, sodium citrate, citric acid, Tris, sodium hydroxide, HCl, or mixtures thereof.

18. The aqueous fulvestrant suspension of claim 14,

wherein the fulvestrant particles:

have one or more of:

an LD Dv(10) between about 1 micron to about 3 microns;

an LD Dv(50) between about 2 microns and about 35 microns; and

an LD Dv(90) between about 4 microns and about 120 microns; or

have one or more of:

an LD Dv(10) between about 1-2 microns;

an LD Dv(50) between about 2-4 microns; and

an LD Dv(90) between about 6-9 microns.

19. The aqueous fulvestrant suspension of claim 14,

wherein the fulvestrant particles:

have one or more of:

a CE Dv(10) between about 4-10 microns,

a CE Dv(50) between about 10-35 microns, and

a CE Dv(90) between about 35-110 microns;

have one or more of:

a CE Dv(10) between about 4-8 microns,

a CE Dv(50) between about 10-25 microns, and

a CE Dv(90) between about 25-60 microns; or

have one or more of:

a CE Dv(10) between about 4-8 microns,

a CE Dv(50) between about 10-20 microns, and

a CE Dv(90) between about 20-35 microns.

20. The aqueous fulvestrant suspension of claim 14,

wherein:

the suspension comprises about 56 mg/mL to about 59 mg/mL of one or more stabilizers; and

the one or more stabilizers comprises:

about 1.0 mg/mL to about 4.0 mg/mL of one or more polyvinylpyrrolidones;

about 5.0 mg/mL of polysorbate 80; and

about 50 mg/mL of mannitol; or

wherein:

the suspension comprises about 56.6 mg/mL to about 57.4 mg/mL of one or more stabilizers; and

the one or more stabilizers comprises:

about 1.6 mg/mL to about 2.4 mg/mL of one or more polyvinylpyrrolidones;

about 5.0 mg/mL of polysorbate 80; and

about 50 mg/mL of mannitol; or

wherein the suspension comprises:

about 1.6 mg/mL to about 2.4 mg/mL of PLASDONE™ C-12 povidone, povidone K12, or a combination thereof;

about 5.0 mg/mL of polysorbate 80; and

about 50 mg/mL of mannitol; or

wherein the suspension comprises:

about 100 mg/mL fulvestrant;

about 1.6 mg/mL of PLASDONE™ C-12 povidone, povidone K12, or a combination thereof;

about 5.0 mg/mL of polysorbate 80; and

about 50 mg/mL of mannitol; or

wherein the suspension comprises:

about 100 mg/mL fulvestrant;

about 2.4 mg/mL of PLASDONE™ C-12 povidone, povidone K12, or a combination thereof;

about 5.0 mg/mL of polysorbate 80; and

about 50 mg/mL of mannitol.

21. The pharmaceutical composition of claim 6,

wherein after the initial administration the additional administration provides:

both a C_maxin the range of 10.04-12.55 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 12.55-15.06 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 15.06-17.57 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 17.57-20.08 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 10.04-11.295 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 11.295-12.55 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 12.55-13.805 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 13.805-15.06 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 15.06-16.315 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 16.315-17.57 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 17.57-18.825 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 18.825-20.08 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 15.06-18.825 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant;

both a C_maxin the range of 12.55-18.825 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant; or

both a C_maxin the range of 10.04-15.06 ng/mL of fulvestrant and an AUC in the range of 9,120-14,250 ng·hr/mL of fulvestrant.

22. The pharmaceutical composition of claim 3, wherein the fulvestrant particles:

have one or more of:

a CE Dv(90) between about 35 microns and about 110 microns;

a CE Dv(50) between about 10 microns and about 35 microns; and

a CE Dv(10) between about 4 microns and about 10 microns;

have one or more of:

a CE Dv(90) between about 25 microns and about 60 microns;

a CE Dv(50) between about 10 microns and about 25 microns; and

a CE Dv(10) between about 4 microns and about 8 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 35 microns;

a CE Dv(50) between about 10 microns and about 20 microns; and

a CE Dv(10) between about 4 microns and about 8 microns;

have one or more of:

a CE Dv(90) between about 30 microns and about 100 microns;

a CE Dv(50) between about 10 microns and about 50 microns; and

a CE Dv(10) between about 4 microns and about 10 microns;

have one or more of:

a CE Dv(90) between about 50 microns and about 100 microns;

a CE Dv(50) between about 20 microns and about 50 microns; and

a CE Dv(10) between about 6 microns and about 8 microns;

have one or more of:

a CE Dv(90) between about 50 microns and about 75 microns;

a CE Dv(50) between about 30 microns and about 40 microns; and

a CE Dv(10) between about 8 microns and about 10 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 60 microns;

a CE Dv(50) between about 9 microns and about 20 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 50 microns;

a CE Dv(50) between about 9 microns and about 20 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 45 microns;

a CE Dv(50) between about 9 microns and about 20 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 40 microns;

a CE Dv(50) between about 9 microns and about 15 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 35 microns;

a CE Dv(50) between about 9 microns and about 15 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have one or more of:

a CE Dv(90) between about 20 microns and about 45 microns;

a CE Dv(50) between about 9 microns and about 15 microns; and

a CE Dv(10) between about 3 microns and about 7 microns;

have a CE Dv(90) between about 10 microns and about 200 microns;

have a CE Dv(50) between about 5 microns and about 60 microns;

have a CE Dv(10) between about 1 microns and about 25 microns;

have one or more of:

a CE Dv(90) between about 10 microns and about 200 microns;

a CE Dv(50) between about 5 microns and about 60 microns; and

a CE Dv(10) between about 1 microns and about 25 microns;

have one or more of:

a CE Dv(90) between about 10 microns and about 200 microns; and

a CE Dv(50) between about 5 microns and about 60 microns;

have one or more of:

a CE Dv(50) between about 5 microns and about 60 microns; and

a CE Dv(10) between about 1 microns and about 25 microns; or

have one or more of:

a CE Dv(90) between about 10 microns and about 200 microns; and

a CE Dv(10) between about 1 microns and about 25 microns.

23. The aqueous fulvestrant suspension of claim 14,

wherein:

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 40% to 80% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 60% to 80% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 50% to 75% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 40% to 50% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 50% to 60% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 60% to 70% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections;

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 70% to 80% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections; or

upon administration of the aqueous fulvestrant suspension to a subject in a single intramuscular injection, the 90% confidence intervals (CI) of the relative mean Cmax of fulvestrant is within 70% to 80% of the relative mean Cmax of fulvestrant after administration of 500 mg of fulvestrant in the form of FASLODEX™ administered intramuscularly as two 5 mL injections.

24. A method of treating a subject having breast cancer, comprising administering to the subject

the suspension of claim 1;

wherein optionally the suspension, pharmaceutical composition, or aqueous fulvestrant suspension comprises about 50 mg/mL or about 100 mg/mL fulvestrant;

wherein optionally the suspension, pharmaceutical composition, or aqueous fulvestrant suspension is administered in combination with one or more additional therapeutic agents, preferably palbociclib.

25. The method of claim 24, wherein the breast cancer is hormone receptor (HR)-positive breast cancer.

26. The method of claim 24, wherein the subject is a post-menopausal human woman with disease progression following antiestrogen therapy.

27. The method of claim 24, wherein the breast cancer is HR-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced or metastatic breast cancer.

28. The method of claim 24, wherein the subject is a human woman with disease progression after endocrine therapy.