WO2023156493A1 - Crystalline forms of naltrexone - Google Patents

Abstract

This disclosure provides crystalline forms of naltrexone, and methods of making and using these forms.

Description

CRYSTALLINE FORMS OF NALTREXONE

FIELD OF THE INVENTION

The present invention relates to novel crystalline forms of naltrexone and their use in pharmaceutical compositions. In particular, an anhydrous form of naltrexone as well as solvated forms are disclosed herein. The invention also relates to methods of use thereof.

BACKGROUND OF THE INVENTION

Alcohol dependence is a prevalent disease with substantial morbidity and mortality. Detoxification and psychosocial therapy provide the basis of treatment; in addition, pharmacotherapy is becoming widely accepted. Administered orally, naltrexone, a potent opioid antagonist, has been shown to reduce relapse to heavy drinking in alcohol dependent patients, decrease the number of drinks consumed when relapse does occur, and promote abstinence. Naltrexone has been reported to reduce both craving and the reinforcing euphoric qualities of alcohol.

Opioid dependence is a chronic disorder that results from a variety of genetic, psychological and environmental factors. Traditional treatment has consisted of two phases: detoxification and rehabilitation. Detoxification ameliorates the symptoms and signs of withdrawal while rehabilitation helps the patient avoid future problems with opioids. In the past, many rehabilitative treatments have been psychosocial. More recently, there has been increasing interest in medication-assisted treatment.

The successful treatment of alcohol dependence or opioid dependence has many serious challenges and complications. Patient compliance can be a particularly difficult challenge to overcome. Accordingly, there is a need for novel and improved therapies.

Polymorphs, solvates and salts of various drugs have been described in the literature as imparting novel properties upon the drug. These crystalline forms can have different solubilities, stabilities and processing characteristics, presenting opportunities and challenges.

Many crystalline forms of naltrexone, along with multiple solvated forms, have been previously disclosed with anhydrous forms being reported (such as US2006/0142320 and US2010/0210675). However, the anhydrous forms reported in the art are thermally liable to solid state conversions within and/or near manufacturing conditions. This can lead to problems with manufacturing naltrexone-containing products. Thus, there is still a need for improved crystalline forms of naltrexone. SUMMARY OF THE INVENTION

Provided herein are crystalline forms useful for the treatment of opioid dependence or alcohol dependence in a subject in need thereof. In an aspect, provided herein are crystalline forms of naltrexone having the formula:

In another aspect, provided herein is a method of treating opioid dependence in a subject in need thereof comprising administering to the subject a crystalline form of naltrexone. In another aspect, provided herein is a method of treating alcohol dependence in a subject in need thereof comprising administering to the subject a crystalline form of naltrexone. In another aspect, provided herein is a method of treating opioid use disorder in a subject in need thereof comprising administering to the subject a crystalline form of naltrexone. In another aspect, provided herein is a method of treating alcohol use disorder in a subject in need thereof comprising administering to the subject a crystalline form of naltrexone.

BRIEF DESCRIPTION OF THE DRAWINGS

The invention is illustrated, but in no way limited, by the tables herein and the following examples, with reference to the figures in which:

FIG. 1 is an experimental PXRD diffractogram for naltrexone anhydrate (Form S).

FIG. 2 is a differential scanning calorimetry (DSC) thermogram for Form S with temperature ramping of 10 degrees C per minute.

FIG. 3 is a differential scanning calorimetry (DSC) thermogram for Form S with temperature ramping of 5 degrees C per minute.

FIG. 4 is an experimental PXRD diffractogram for naltrexone sesquiacetonitrile solvate (Form T). FIG. 5 is an experimental PXRD diffractogram for naltrexone acetonitrile solvate (Form U).

DETAILED DESCRIPTION OF THE INVENTION

Applicants surprisingly discovered novel naltrexone crystalline forms, including solvates and anhydrous forms. Applicants appreciate that the novel crystalline forms of naltrexone, for example, the anhydrate form of naltrexone, have superior properties for use in naltrexone-containing compositions relative to other known forms of naltrexone.

Pharmaceutical compositions containing the naltrexone crystalline forms described herein, when formulated for administration, are useful in the treatment and prevention of, for example, opioid use disorder, opioid dependence, alcohol use disorder and alcohol dependence, as well as other naltrexone-based therapies.

The solid state of a compound can be important when the compound is used for pharmaceutical purposes. The physical properties of a compound can change from one solid form to another, which can affect the suitability of the form for pharmaceutical use. For example, a particular crystalline solid compound can overcome the disadvantage of other solid forms of the compound such as, e.g., physical instability and/or reduced purity.

As with all pharmaceutical compounds and compositions, the chemical and physical properties of the naltrexone form(s) utilized can be important in commercial development and manufacturing. These properties include, but are not limited to: (1) packing properties such as molar volume, density and hygroscopicity, (2) thermodynamic properties such as melting temperature, vapor pressure and solubility, (3) kinetic properties such as dissolution rate and stability (including stability at ambient conditions, especially to moisture, and under storage conditions), (4) surface properties such as surface area, wettability, interfacial tension and shape, (5) mechanical properties such as hardness, tensile strength, compactibility, handling, flow and blend; and (6) filtration properties. These properties can affect, for example, processing and storage of pharmaceutical compositions comprising naltrexone. Solid state forms of naltrexone that provide an improvement in one or more of these properties relative to other solid state forms of naltrexone are desirable. For example, the naltrexone anhydrate Form S described herein possesses improved hygroscopicity relative to other previously disclosed forms of naltrexone anhydrate.

The crystalline forms of the invention and the compositions containing them have the advantage that they are in a form which provides for improved ease of handling and/or processability. Further, depending upon the intended use, they have improved chemical and solid state stability. For example, they may be stable when stored over prolonged periods of time. They may be prepared in good yields, in higher purity, in less time, more conveniently and at a lower cost, than forms of naltrexone prepared previously.

Provided herein are solid, crystalline forms of naltrexone. In particular, provided herein, are crystalline forms of anhydrous naltrexone and crystalline forms of naltrexone solvate.

The crystalline forms provided herein can be characterized by powder X-ray diffraction (PXRD) and/or differential scanning calorimetry (DSC).

In one embodiment, the crystalline form of the anhydrate (Form S) disclosed herein is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, and 13.24.

In another embodiment, the crystalline form of the sesquiacetonitrile solvate (Form T) disclosed herein is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80, and 12.20.

In another embodiment, the crystalline form of the acetonitrile solvate (Form U) disclosed herein is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 6.13, 8.08 and 8.66.

Powder X-Ray Diffraction

Most of the various crystalline forms of naltrexone were analyzed using Powder x- Ray Diffraction. Powder x-ray diffraction (PXRD) diffractograms can be obtained using, for example, a PANalytical X’Pert PRO MPD diffractometer using an incident beam of Cu radiation produced using an Optix long, fine-focus source. An elliptically graded multilayer mirror was used to focus Cu Kα x-rays through the specimen and onto the detector.

PXRD diffractograms of the various naltrexone materials obtained by the methods described herein are shown in the Examples and Figures. The naltrexone forms of the invention are not limited to those made in accordance with the methods described herein.

Single Crystal X-Ray Diffraction

Samples of single crystals can be obtained and analyzed. Single crystal x-ray diffraction data was obtained using a Rigaku Supernova diffractometer, equipped with a copper anode microfocus sealed x-ray tube (Cu Kα = 1.54184 Å). Those skilled in the art will recognize other appropriate means of collecting single-crystal x-ray diffraction data. Melting/Decomposition Temperature

The temperatures of melting and/or decomposition of naltrexone crystalline forms were determined using differential scanning calorimetry (DSC). DSC measurements can be obtained, for example, on a TA Instruments DSC2500 Discovery Series thermal analysis system. Samples of approximately 1-2 mg can be accurately weighed in an aluminum DSC pan and covered with an aluminum lid that was crimped in place. The samples can then be heated over the range of 25-190 degrees C., for example, at a heating rate of 10 degrees C./min.

Melting/decomposition temperature ranges were defined from the extrapolated onset to the maximum of the melting/decomposition endotherm.

Those skilled in the art will recognize other appropriate means of measuring DSC.

Definitions

Listed below are definitions of various terms used to describe the crystalline forms provided herein. These definitions apply to the terms as they are used throughout this specification and claims, unless otherwise limited in specific instances, either individually or as part of a larger group.

Unless defined otherwise, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which the compound and its crystalline forms belong.

As used herein, the articles “a” and “an” refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

As used herein, “addiction” is generally defined as a chronic brain disease that causes compulsive drug seeking and use, or alcohol seeking and use. Drug addiction can be opioid addiction (i.e., opioid dependence), stimulant addiction, and the like.

As used herein, the term “pharmaceutically acceptable carrier” refers to a pharmaceutically acceptable material, composition or carrier, such as a liquid or solid filler, stabilizer, dispersing agent, suspending agent, diluent, excipient, thickening agent, solvent or encapsulating material, involved in carrying or transporting a compound useful within the present disclosure within or to the patient such that it may perform its intended function. Typically, such constructs are carried or transported from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation, including the compound provided herein, and not injurious to the patient.

Some examples of materials that may serve as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; surface active agents; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; phosphate buffer solutions; and other non-toxic compatible substances employed in pharmaceutical formulations.

As used herein, “pharmaceutically acceptable carrier” also includes any and all coatings, antibacterial and antifungal agents, and absorption delaying agents, and the like that are compatible with the activity of the compound provided herein, and are physiologically acceptable to the patient. Supplementary active compounds may also be incorporated into the compositions. Other additional ingredients that may be included in the pharmaceutical compositions used in the practice of the present disclosure are known in the art and described, for example in Remington's Pharmaceutical Sciences (Genaro, Ed., Mack Publishing Co., 1985, Easton, PA), which is incorporated herein by reference.

As used herein, the phrases “therapeutically effective dose” and “therapeutically effective amount” refer to an amount of a compound that prevents the onset, alleviates the symptoms, stops the progression of a disease, or results in another desired biological outcome such as, e.g., improved clinical signs.

As used herein, the term “treat,” “treated,” “treating,” or “treatment” includes the diminishment or alleviation of at least one symptom associated or caused by the state, disorder or disease being treated. In certain embodiments, the treatment comprises bringing into contact with the opioid receptor an effective amount of the compound provided herein for conditions related to opioid dependence, alcohol dependence or addiction.

As used herein, the term “prevent” or “prevention” means no disorder or disease development if none had occurred, or no further disorder or disease development if there had already been development of the disorder or disease. Also considered is the ability of one to prevent some or all of the symptoms associated with the disorder or disease. As used herein, the term “patient,” “individual” or “subject” refers to a human or a non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. In an embodiment, the patient, subject, or individual is human.

The term “administering “or “administration” and the like, refers to providing a therapeutic agent, such as a crystalline form disclosed herein, to the subject in need of treatment. In an embodiment, the subject is a mammal. In another embodiment, the subject is a human.

As used herein, the term “about” will be understood by persons of ordinary skill in the art and will vary to some extent on the context in which it is used. As used herein when referring to a measurable value such as an amount, a temporal duration, and the like, the term “about” is meant to encompass variations of ±20% or ±10%, including ±5%, ±1%, and ±0.1% from the specified value, as such variations are appropriate to perform the disclosed methods.

Characterization of Crystalline Forms

In certain embodiments, the crystalline forms described herein are identifiable on the basis of characteristic peaks in a powder X-ray diffraction analysis. Powder X-ray diffraction (PXRD) is a scientific technique using X-ray, neutron, or electron diffraction on powder, microcrystalline, or other solid materials for structural characterization of solid materials. A description of the methods used to obtain certain PXRD diffractograms in connection with the crystalline forms provided herein can be found in the Examples and specification. The PXRD data provided herein is obtained by a method utilizing Cu Kα radiation.

Naltrexone Anhydrate (Form S)

In an aspect, provided herein is the crystalline form naltrexone anhydrate (Form S).

In one embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) as shown in FIG. 1.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) as shown in Table 1. In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59 and 13.24.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, 12.21, 13.24, 15.10, and 15.48.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, 11.32, 12.21, 13.24, 14.13, 15.10, 15.48, and 16.19.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 11.32, 13.24, 15.10 and 17.35.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 11.32, 13.81, 15.48 and 17.35.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, 11.32, 12.21 and 13.81.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 10.59, 11.32, 13.24, 15.10 and 17.35.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by an PXRD diffractogram having peaks expressed in degrees 2-theta at angles (±0.2 degrees) of:

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) is characterized by any two, three, four, five, six, seven, or eight peaks from the list above.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) has a PXRD diffractogram substantially as depicted in FIG. 1.

In another embodiment, the crystalline form of the naltrexone anhydrate (Form S) has a DSC thermogram characterized by an endotherm with an onset temperature of 174 degrees C.

Two anhydrate crystalline forms of naltrexone were previously described in the art. US2006/0142320 (Alkermes, Inc.) describes a crystalline anhydrate (referred to herein as “Form A”). US2010/0210675 (Cilag AG) describes a solvent-free crystalline form (referred to herein as “Form K”).

Naltrexone Sesquiacetonitrile Solvate (Form T)

In another aspect, provided herein is the crystalline form naltrexone sesquiacetonitrile solvate (Form T).

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) as shown in FIG. 4.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) as shown in Table 3. In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80 and 12.20.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 12.20, 15.05, 15.59 and 16.65.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80, 12.20, 15.05, 15.59, 16.65, 20.01, 21.31 and 22.96.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 15.05, 15.59 and 16.65.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80, 13.81 and 15.05.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 9.80, 12.20, 18.17 and 20.01.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 15.05, 16.65 and 18.49.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile solvate (Form T) is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80, 12.20 and 15.59.

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile (Form T) is characterized by an PXRD diffractogram having peaks expressed in degrees 2- theta at angles (±0.2 degrees) of:

In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile (Form T) is characterized by any two, three, four, five, six, seven, or eight peaks from the list above. In another embodiment, the crystalline form of the naltrexone sesquiacetonitrile

(Form T) has an PXRD diffractogram substantially as depicted in FIG. 4. Naltrexone Acetonitrile Solvate (Form U)

In another aspect, provided herein is the crystalline form naltrexone acetonitrile solvate (Form U).

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) as shown in FIG. 5.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) as shown in Table 5.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 6.13, 8.08 and 8.66.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 6.13, 8.08, 9.12, 10.62 and 13.03.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 6.13, 8.08, 8.66, 9.12, 10.11, 10.62, 12.60, 13.03 and 14.79.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 8.08, 9.12, 12.60 and 13.03.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 8.08, 8.66, 9.12 and 10.62.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 6.13, 8.66, 13.03, 14.79 and 15.06.

In another embodiment, the crystalline form of the naltrexone acetonitrile solvate (Form U) is characterized by a PXRD diffractogram having peaks expressed in degrees 2- theta at angles (± 0.2 degrees) of 6.13, 9.12, 10.62 and 12.60.

In another embodiment, the crystalline form of the naltrexone acetonitrile (Form U) is characterized by an PXRD diffractogram having peaks expressed in degrees 2-theta at angles (±0.2 degrees) of:

In another embodiment, the crystalline form of the naltrexone acetonitrile (Form U) is characterized by any two, three, four, five, six, seven, or eight peaks from the list above.

In another embodiment, the crystalline form of the naltrexone acetonitrile (Form U) has the PXRD diffractogram substantially as depicted in FIG. 5.

Methods of Treatment

Provided herein are methods for the treatment of a disease comprising administering a crystalline form of naltrexone as disclosed herein, or a pharmaceutical composition comprising the crystalline form and a pharmaceutically acceptable carrier.

In a non-limiting aspect, naltrexone modulates the p-opioid receptor. In a particular embodiment, naltrexone is a p-receptor antagonist. As such, in one aspect, naltrexone is useful in the treatment of opioid dependence or alcohol dependence in a subject by acting as an antagonist of the p-receptor.

In another embodiment, the crystalline forms disclosed herein can be used to treat addiction in a subject in need thereof. The addiction can be drug addiction or alcohol addiction.

The drug addiction can be one or more of opioid addiction (i.e., opioid dependence), opioid use disorder, or stimulant addiction. The opioid can be one or more of fentanyl, morphine, oxymorphone, buprenorphine, hydromorphone, oxycodone, hydrocodone, or the like. The drug addiction can also be one or more of diamorphine (i.e., heroin), ***e, nicotine, and amphetamine.

In one embodiment, the crystalline forms disclosed herein can be used to treat a disease or condition in a subject, wherein the subject has a tolerance to opioid medication, the subject has a history of opioid dependency or abuse, the subject is at risk of opioid dependency or abuse, or in circumstances wherein it is desirable that the risk of opioid dependence, opioid addiction, or symptoms of opioid withdrawal in the subject is minimized.

The crystalline forms disclosed herein can also be used to treat alcohol use disorder, alcohol dependence, or alcohol addiction, which can also be referred to as alcoholism. “Alcoholism” refers to an addictive disease or disorder characterized by an inability to control the intake of alcohol, i.e., a continued excessive or compulsive use of alcoholic drinks. Alcoholism may involve changes an individual’s ability to metabolize alcohol as well. Diagnosis of alcoholism can be made by psychiatric examination.

In one embodiment of the methods described herein, the subject is human.

Pharmaceutical Compositions

In an aspect, provided herein is a pharmaceutical composition comprising a crystalline form as disclosed herein and a pharmaceutically acceptable carrier.

In an embodiment, the pharmaceutical composition comprises a crystalline form that is substantially free from other crystalline forms.

The pharmaceutical compositions can be formulated for oral, intravenous, intramuscular, subcutaneous or parenteral administration for the therapeutic or prophylactic treatment of opioid dependence, alcohol dependence, opioid use disorder, alcohol use disorder or relapse to opioid dependence.

The pharmaceutical preparations disclosed herein can be prepared in accordance with standard procedures and are administered at dosages that are selected to reduce, prevent or eliminate disease. See, for example, Remington’s Pharmaceutical Sciences, Mack Publishing Company, Easton, PA and Goodman and Gilman’s “The Pharmaceutical Basis of Therapeutics,” Pergamon Press, New York, NY, the contents of which are incorporated herein by reference, for a general description of the methods for administering various agents for human therapy.

The pharmaceutical compositions described herein can comprise a crystalline form disclosed herein in association with one or more nontoxic, pharmaceutically acceptable carriers and/or diluents and/or adjuvants and/or excipients.

For oral or parenteral administration, the crystalline form disclosed herein can be mixed with conventional pharmaceutical carriers and excipients and used in the form of tablets, capsules, elixirs, suspensions, syrups, wafers and the like. The compositions comprising a crystalline form disclosed herein can contain from about 0.1% to about 99% by weight of the active compound, such as from about 10% to about 30%.

For oral use, solid formulations such as tablets and capsules are useful. Sustained release or enterically coated preparations can also be devised. For pediatric and geriatric applications, one embodiment provides suspensions, syrups and chewable tablets. For oral administration, the pharmaceutical compositions are in the form of, for example, a tablet, capsule, suspension or liquid. The pharmaceutical compositions can be made in the form of a dosage unit containing a therapeutically-effective amount of the active ingredient. Examples of such dosage units are tablets and capsules. For therapeutic purposes, the tablets and capsules which can contain, in addition to the active ingredient, conventional carriers such as binding agents, fillers, lubricants, disintegrants, or acceptable wetting agents. Oral liquid preparations generally are in the form of aqueous or oily solutions, suspensions, emulsions, syrups or elixirs.

The pharmaceutical compositions disclosed herein can be placed in a pharmaceutically acceptable carrier and are delivered to a recipient subject (e.g., a human) in accordance with known methods of drug delivery. In general, the methods of delivering the pharmaceutical compositions in vivo utilize art-recognized protocols for delivering the agent with the only substantial procedural modification being the substitution of a crystalline form of the present disclosure for the drugs in the art-recognized protocols.

Administration / Dosage / Formulations

In another aspect, provided herein is a pharmaceutical composition comprising a crystalline form provided herein, together with a pharmaceutically acceptable carrier.

Actual dosage levels of the active ingredients in the pharmaceutical compositions discussed herein may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

In particular, the selected dosage level will depend upon a variety of factors including the activity of the particular compound employed, the time of administration, the rate of excretion of the compound, the duration of the treatment, other drugs, compounds or materials used in combination with the crystalline form, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well, known in the medical arts.

A medical doctor, e.g., physician or veterinarian, having ordinary skill in the art may readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could begin administration of the pharmaceutical composition to dose the disclosed crystalline form at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

In particular embodiments, it is especially advantageous to formulate the crystalline form in dosage unit form for ease of administration and uniformity of dosage. “Dosage unit form,” as used herein, refers to physically discrete units suited as unitary dosages for the patients to be treated; each unit containing a predetermined quantity of the disclosed compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical vehicle. The dosage unit forms of the crystalline form disclosed herein are dictated by and directly dependent on (a) the unique characteristics of the disclosed compound and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compounding/formulating such a disclosed compound for the treatment of opioid dependence, alcohol dependence, opioid use disorder, alcohol use disorder, or drug addiction in a patient.

In one embodiment, the crystalline form provided herein is formulated using one or more pharmaceutically acceptable excipients or carriers. In one embodiment, the pharmaceutical compositions comprise a therapeutically effective amount of the disclosed crystalline form and a pharmaceutically acceptable carrier.

In some embodiments, the dose of a disclosed compound is from about 1 mg to about 1,000 mg. In some embodiments, a dose of the disclosed compound used in compositions described herein is less than about 1,000 mg, or less than about 800 mg, or less than about 600 mg, or less than about 500 mg, or less than about 300 mg, or less than about 200 mg, or less than about 100 mg, or less than about 50 mg, or less than about 20 mg, or less than about 10 mg. For example, a dose is about 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 120 mg, 140 mg, 160 mg, 180 mg, 200 mg, 220 mg, 240, 260 mg, 280 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, or about 600 mg.

Routes of administration of any of the compositions disclosed herein include oral, nasal, rectal, intravaginal, parenteral, buccal, sublingual or topical. The compound for use provided herein may be formulated for administration by any suitable route, such as for oral or parenteral, for example, transdermal, transmucosal (e.g., sublingual, lingual, (trans)buccal, (trans)urethral, vaginal (e.g., trans- and perivaginally), (intra)nasal and (trans)rectal), intravesical, intrapulmonary, intraduodenal, intragastrical, intrathecal, subcutaneous, intramuscular, intradermal, intra-arterial, intravenous, intrabronchial, inhalation, and topical administration. In one embodiment, the preferred route of administration is oral.

Suitable compositions and dosage forms include, for example, tablets, capsules, caplets, pills, gel caps, troches, dispersions, suspensions, solutions, syrups, granules, beads, transdermal patches, gels, powders, pellets, magmas, lozenges, creams, pastes, plasters, lotions, discs, suppositories, liquid sprays for nasal or oral administration, dry powder or aerosolized formulations for inhalation, compositions and formulations for intravesical administration and the like. It should be understood that the formulations and compositions that would be useful in the present disclosure are not limited to the particular formulations and compositions that are described herein.

For oral application, particularly suitable are tablets, dragees, liquids, drops, suppositories, or capsules, caplets and gelcaps. The compositions intended for oral use may be prepared according to any method known in the art and such compositions may contain one or more agents selected from the group consisting of inert, non-toxic pharmaceutically excipients that are suitable for the manufacture of tablets. Such excipients include, for example, an inert diluent such as lactose; granulating and disintegrating agents such as cornstarch; binding agents such as starch; and lubricating agents such as magnesium stearate. The tablets may be uncoated or they may be coated by known techniques for elegance or to delay the release of the active ingredients. Formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert diluent.

For parenteral administration, the disclosed compound may be formulated for injection or infusion, for example, intravenous, intramuscular or subcutaneous injection or infusion, or for administration in a bolus dose or continuous infusion. Suspensions, solutions or emulsions in an oily or aqueous vehicle, optionally containing other formulatory agents such as suspending, stabilizing or dispersing agents may be used.

It has been discovered that controlling the crystallinity of the total amount of naltrexone can have a substantial impact upon the duration of release. For example, a composition containing PLGA (i.e., poly(lactic-co-glycolic acid)) microspheres characterized by total % naltrexone crystallinity in PLGA can impact the release profile of naltrexone upon parenteral administration. Alternatively, instead of incorporating naltrexone into polymeric particles, it is possible to entrap these materials in microparticles prepared. For example, coacervation techniques, interfacial polymerization (for example, hydroxymethylcellulose or gelatine-microcapsules and poly-(methylmethacrylate) microcapsules, respectively), colloidal drug delivery systems (for example, liposomes, albumin, microparticles, microemulsions, nanoparticles, and nanocapsules), or macroemulsion systems can be used.

Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, numerous equivalents to the specific procedures, embodiments, claims, and examples described herein. Such equivalents were considered to be within the scope of this disclosure and covered by the claims appended hereto. For example, it should be understood, that modifications in reaction conditions, including but not limited to reaction times, reaction size/volume, and experimental reagents, such as solvents, catalysts, pressures, atmospheric conditions, e.g., nitrogen atmosphere, and reducing/oxidizing agents, with art- recognized alternatives and using no more than routine experimentation, are within the scope of the present application.

It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present disclosure. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application.

The following examples further illustrate aspects of the present disclosure. However, they are in no way a limitation of the teachings of the present disclosure as set forth.

EXEMPLIFICATION

The crystalline forms of the invention were made as described below. Thereafter the resulting material was analyzed using the analytical techniques described herein, that is, powder X-ray diffraction, differential scanning calorimetry, and single crystal x-ray diffraction.

Analytical Methods

Single-crystal x-ray diffraction (SCXRD) was performed with a Rigaku SuperNova diffractometer, equipped with a copper anode microfocus sealed X-ray tube (Cu Kα λ = 1.54184 Å) and a Dectris Pilatus3 R 200K hybrid pixel array detector.

PXRD was performed with a PANalytical X’Pert PRO MPD diffractometer using an incident beam of Cu Kα radiation produced using a long, fine-focus source and a nickel filter. Diffraction patterns were collected using a scanning position-sensitive detector (X’Celerator) located 240 mm from the sample and Data Collector software v. 5.5.

The relative intensity of each diffractogram peak in Tables 1, 3 and 5, as well as FIGS. 1, 4 and 5, may change or shift under certain conditions, although the crystalline form is the same. One of ordinary skill in the art should be able to readily determine whether a given crystalline form is the same crystalline form as described in one of FIGS. 1, 4 or 5 or Tables 1, 3 and 5.

DSC was performed using a DSC2500 Discovery Series differential scanning calorimeter. The sample was placed into a hermetically sealed aluminum DSC pan, the weight was accurately recorded, and the sample was inserted into the DSC cell. The samples were analyzed from 25 degrees C to 190 degrees C at 10 degrees C/minute, unless otherwise indicated.

Example 1- Naltrexone Anhydrate Form S

The anhydrate Form S was obtained according to the following method: A 20 mL vial was charged with 76.6 mg of commercially available naltrexone base anhydrous (Mallinckrodt; lot 1111000327; referred to herein as “Form A”) and placed into a preheated 165 degrees C oven. The sample was removed after 25 minutes and cooled to room temperature. Upon cooling, the sample was noted to be a light brown cracked glass containing crystals dispersed throughout. The crystals were collected and characterized using PXRD and DSC.

The anhydrate Form S was also, independently, obtained by heating 2.4 grams of commercially available naltrexone base anhydrous (sourced from Aesica, lot 6072807; mixture of Form A and Form K) at 125 degrees C for 24 hours. Full conversion to Form S was determined by PXRD.

Form S exhibits a higher melt onset and appears more thermodynamically stable relative to either Form A or Form K, both of which are also anhydrous forms of naltrexone, at all temperatures of practical importance.

The PXRD diffractogram for the anhydrate Form S provided herein is shown in Figure 1. Table 1 also shows PXRD data for a sample of naltrexone anhydrate Form S.

Table 1

DSC analysis of the anhydrate Form S was performed by ramping 10 degrees C per minute from ambient temperature (about 20 degrees C) to 190 degrees C. The DSC thermogram is depicted in Figure 2. An endotherm is observed with an onset temperature of 173.5 degrees C and a peak temperature of 175.4 degrees C.

DSC analysis of the anhydrate Form S was also performed by ramping 5 degrees C per minute from ambient temperature (about 20 degrees C) to 190 degrees C. The DSC thermogram is depicted in Figure 3. An endotherm is observed with an onset temperature of 174.1 degrees C and a peak temperature of 175.4 degrees C.

Example 2- Anhydrate Form S anhydrate Single Crystal Data

A single crystal of anhydrate Form S was separated from the discolored melt of Form K (described as solvent-free crystalline form in US2010/0210675 by Cilag AG), achieved by exposing Form A to 165 degrees C for 20 minutes. The resulting material was then allowed to cool to room temperature, resulting in large crystals dispersed in cooled melt (glass). A suitable single crystal was selected and analyzed by SCXRD. The crystal system is monoclinic and the space group is 21. The cell parameters and calculated volume are: a = 12.61399(10) Å, b = 22.35431(19) Å, c = 12.63774(11) Å, α = 90°, β = 97.4775(8)°, y = 90°, V= 3533.25(5) Å3. The molecular weight is 341.39 g mol-1 with Z = 8, resulting in a calculated density of 1.284 g cm^-3. Further details of the crystal data and crystallographic data collection parameters are summarized in Table 2.

Table 2 Anhydrate Form S Crystal Data and Collection Parameters

Empirical formula: C20H23NO4 Formula weight (g mol^-1): 341.39 Temperature (K): 299.7(2) Wavelength (Å): 1.54184

Crystal system: monoclinic

Space group: F21

Unit cell parameters: a = 12.61399(10) Å; a = 90° b = 22.35431(19) Å; p = 97.4775(8)° c = 12.63774(11) Å; y = 90°

Unit cell volume (ų): 3533.25(5)

Cell formula units, Z: 8

Calculated density (g cm^-3): 1.284

Absorption coefficient (mm^-1): 0.725

F(000): 1456

Crystal size (mm³): 0.23 x 0.09 x 0.08

Reflections used for cell measurement: 20362

Q range for cell measurement: 3.9450°-77.2250°

Total reflections collected: 38749

Index ranges: -14 ≤ h ≤ 15; -28 ≤ k ≤ 27; -15 ≤ I ≤ 16

0 range for data collection: θmin = 3.527°, θmax = 77.730°

Completeness to θmax : 98.9%

Completeness to θfull = 67.684°: 100%

Absorption correction: multi-scan

Transmission coefficient range: 0.786-1.000

Refinement method: full matrix least-squares on F²

Independent reflections: 14557 [Rint = 0.0238, Rσ = 0.0262]

Reflections [ />2σ(/) ]: 12977

Reflections / restraints / parameters: 14557 / 31 / 1030

Goodness-of-fit on F²: S = 1.04

Final residuals [ />2σ(/) ] : R = 0.0445, R_w = 0.1176

Final residuals [ all reflections ]: R = 0.0498, R_w = 0.1225

Largest diff peak and hole (e Å^-3): 0.347, -0.251

Max/mean shift/standard uncertainty: 0.000 / 0.000

Absolute structure determination: Flack parameter: -0.06(5).

Example 3- Sesquiacetonitrile Solvate (Form T) The sesquiacetonitrile solvate (Form T) was obtained according to the following method: A vial was charged with 138 mg of commercially available naltrexone base anhydrous (Aesica; lot 6072807) and contacted with 0.5 mL acetonitrile. The sample was warmed on an 85 degrees C hot plate and removed once visual confirmation of near complete dissolution noted. Upon removal, nucleation was immediately observed. The solution was decanted and the crystals were left wet. The isolated material was observed to expel solvent and a morphology change noted, indicative of a partial desolvation and form conversion.

The PXRD diffractogram for the sesquiacetonitrile solvate (Form T) provided herein is shown in Figure 4. Table 3 also shows PXRD data for a sample of naltrexone sesquiacetonitrile solvate (Form T).

Table 3

Example 4- Sesquiacetonitrile solvate (Form T) Single Crystal Data

The crystal system is orthorhombic and the space group is P212121. The cell parameters and calculated volume are: a = 12.53697(11) Å, b = 12.64062(11) Å, c = 26.0774(2) Å, α = 90°, β = 90°, y = 90°, V= 4132.62(6) Å3. The formula weight is 402.97 g mol-1 with Z = 8, resulting in a calculated density of 1.295 g cm-3. Further details of the crystal data and crystallographic data collection parameters are summarized in Table 4. The asymmetric unit shown contains two naltrexone molecules and three acetonitrile molecules.

Table 4 Sesquiacetonitrile Solvate (Form T) Crystal Data and Collection Parameters

Empirical formula: C23H27.50N2.50O4

Formula weight (g mol^-1): 402.97

Temperature (K): 150.01(10)

Wavelength (Å): 1.54184

Crystal system: orthorhombic

Space group: 212121

Unit cell parameters: a = 12.53697(11) Å; α = 90° b = 12.64062(11) Å; β = 90° c = 26.0774(2) Å; y = 90°

Unit cell volume (ų): 4132.62(6)

Cell formula units, Z: 8

Calculated density (g cm^-3): 1.295 Absorption coefficient (mm^-1 ): 0.723

F(000): 1720

Crystal size (mm³): 0.48 x 0.36 x 0.2

Reflections used for cell measurement: 17450

Q range for cell measurement: 3.8670°-77.2660°

Total reflections collected: 21844

Index ranges: -15 ≤ h ≤ 15; -14 < k ≤ 15; -32 ≤ I ≤ 30 0 range for data collection: min = 3.886°, θmax = 77.845° Completeness to θmax : 98.7%

Completeness to θfull = 67.684°: 100%

Absorption correction: multi-scan

Transmission coefficient range: 0.829-1.000

Refinement method: full matrix least-squares on F²

Independent reflections: 8547 [Rint = 0.0247, Rσ = 0.0260]

Reflections [ I>2σ (I) ]: 8278

Reflections / restraints / parameters: 8547 / 0 / 551 Goodness-of-fit on F²: S = 1.05

Final residuals [ />2σ(/) ] : R = 0.0449, Rw = 0.1254

Final residuals [ all reflections ]: R = 0.0461, Rw = 0.1265

Largest diff peak and hole (e Å^-3): 0.658, -0.413

Max/mean shift/standard uncertainty: 0.000 / 0.000

Absolute structure determination: Flack parameter: -0.02(6).

Example 5- Acetonitrile Solvate (Form U)

The acetonitrile solvate (Form U) was obtained according to the following method: A vial was charged with 110.8 mg of commercially available naltrexone base anhydrous (Aesica; lot 6072807) and contacted with 0.5 ml of acetonitrile. The sample was heated briefly on an 85 degrees C hot plate until dissolution occurred. The solution was transferred to vial and placed onto a programmable heating/cooling orbital shaker preset at 60 degrees C. A cooling/heating cycling profile was utilized. The cycling profile utilized an initial 60 degrees C hold time (held for 3 hours) followed by a cooling step from 60 degrees C to 15 degrees C (held for 3 hours) then heated to 60 degrees C (held for 3 hours). The cycling process ran for a total of 5 days. Solids were observed to form after 1.5 hours during the initial 3 hour, 60 degrees C set point.

The PXRD diffractogram for the acetonitrile solvate (Form U) provided herein is shown in Figure 5. Table 5 also shows PXRD data for a sample of naltrexone acetonitrile solvate (Form U).

Table 5

While this invention has been particularly shown and described with references to preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

Claims

What is claimed is:

1. A crystalline form of naltrexone anhydrate Form S.

2. The crystalline form of claim 1, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59 and 13.24.

3. The crystalline form of claim 1 or 2, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 11.32, 13.24, 15.10 and 17.35.

4. The crystalline form of any one of claims 1-3, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, 11.32, 12.21 and 13.81.

5. The crystalline form of any one of claims 1-4, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 10.59, 11.32, 12.21, 13.24, 14.13, 15.10, 15.48, and 16.19.

6. The crystalline form of claim 1, wherein the crystalline form is characterized by the PXRD diffractogram substantially as depicted in FIG. 1.

7. The crystalline form of any one of claims 1-6 having a DSC thermogram characterized by an endotherm with an onset temperature of 174 degrees C.

8. The crystalline form of any one of claims 1-7 having a DSC thermogram characterized by an endothermic peak temperature at 175 degrees C.

9. A crystalline form of naltrexone sesquiacetonitrile solvate Form T.

10. The crystalline form of claim 9, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80 and 12.20.

11. The crystalline form of claim 9 or 10, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 15.05, 15.59 and 16.65.

12. The crystalline form of any one of claims 9-11, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 9.80, 12.20, 18.17 and 20.01.

13. The crystalline form of any one of claims 9-11, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 7.67, 9.80, 12.20, 15.05, 15.59, 16.65, 20.01, 21.31 and 22.96.

14. The crystalline form of claim 9, wherein the crystalline form is characterized by the PXRD diffractogram substantially as depicted in FIG. 4.

15. A crystalline form of naltrexone acetonitrile solvate Form U.

16. The crystalline form of claim 15, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 6.13, 8.08 and 8.66.

17. The crystalline form of claim 15 or 16, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 8.08, 9.12, 12.60 and 13.03.

18. The crystalline form of any one of claims 15-17, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 6.13, 9.12, 10.62 and 12.60.

19. The crystalline form of any one of claims 15-17, wherein the crystalline form is characterized by a PXRD diffractogram having peaks expressed in degrees 2-theta at angles (± 0.2 degrees) of 6.13, 8.08, 8.66, 9.12, 10.11, 10.62, 12.60, 13.03 and 14.79.

20. The crystalline form of claim 15, wherein the crystalline form is characterized by the PXRD diffractogram substantially as depicted in FIG. 5.

21. A pharmaceutical composition comprising a crystalline form of any one of claims 1- 20 and a pharmaceutically acceptable carrier.

22. A method of treating opioid dependence in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a crystalline form of any one of claims 1 to 20 or a pharmaceutical composition of claim 21.

23. A method of treating alcohol dependence in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a crystalline form of any one of claims 1 to 20 or a pharmaceutical composition of claim 21.

24. A method of preventing relapse to opioid dependence in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a crystalline form of any one of claims 1 to 20 or a pharmaceutical composition of claim 21.

25. A method of treating alcohol use disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a crystalline form of any one of claims 1 to 20 or a pharmaceutical composition of claim 21.

26. Use of a crystalline form of any one of claims 1 to 20 for the manufacture of a medicament for opioid dependence.

27. Use of a crystalline form of any one of claims 1 to 20 for the manufacture of a medicament for alcohol dependence.

28. Use of a crystalline form of any one of claims 1 to 20 for the manufacture of a medicament for preventing relapse to opioid dependence.

29. Use of a crystalline form of any one of claims 1 to 20 for the manufacture of a medicament for alcohol use disorder.

30. A crystalline form of any one of claims 1 to 20 for use in a method of treating opioid dependence in a subject in need thereof.

31. A crystalline form of any one of claims 1 to 20 for use in a method of treating alcohol dependence in a subject in need thereof.

32. A crystalline form of any one of claims 1 to 20 for use in a method of preventing relapse to opioid dependence in a subject in need thereof.

33. A crystalline form of any one of claims 1 to 20 for use in a method of treating alcohol use disorder in a subject in need thereof.