Novel formulations for treating copper metabolism-related diseases or disorders

Abstract

The present disclosure relates to novel formulations of bis-choline tetrathiomolybdate useful for treating copper metabolism-related diseases or disorders, such as Wilson's disease (WD). For example, the present disclosure relates to low-dose formulations of bis-choline tetrathiomolybdate.

Description

[Technical field]

[0001] The present disclosure relates to novel formulations of bis-choline tetrathiomolybdate useful for treating copper metabolism-related diseases or disorders, such as Wilson's disease (WD). For example, the present disclosure relates to low-dose formulations, such as minitablets, of bis-choline tetrathiomolybdate, as well as capsules, sachets, and kits containing these formulations. [Background technology]

[0002] Wilson's disease (WD) is a rare autosomal recessive disorder of copper (Cu) transport deficiency that results in pathological copper accumulation. In WD, mutations in the ATP7B gene result in defective production of adenosine triphosphatase 2 (ATPase2), which in turn results in impaired biliary excretion of Cu and impaired incorporation of Cu into serum ferroxidase, ceruloplasmin (CP), which contains more than 95% of the Cu found in plasma in healthy humans. This results in increased Cu in the liver, brain, and other tissues, resulting in organ damage and dysfunction. The initial signs and symptoms of WD are primarily hepatic, neurological, or psychiatric disorders, but patients often develop both hepatic and neuropsychiatric disorders. Untreated or inadequately treated patients have progressive disease and usually die from cirrhosis of the liver. Other causes of death associated with WD include hepatic malignancies and worsening neurological symptoms with severe debilitation.

[0003] Current treatments for WD are general chelator therapy, where D-penicillamine and trientine chelate Cu to promote urinary Cu excretion, and zinc (Zn) blocks dietary intake of Cu by upregulating intestinal metallothionein. Currently available drugs are associated with high rates of treatment discontinuation due to tolerability and efficacy issues, as well as non-adherence to treatment regimens. For example, currently available drugs require frequent dosing (e.g., 2-4 times per day) and must be taken in a fasting state for each dose. Their adverse event (AE) profile and complex dosing regimens lead to poor treatment adherence and high treatment failure rates, which are major concerns in WD requiring lifelong treatment.

[0004] Bis-choline tetrathiomolybdate (BC-TTM, also known as thiomolybdate choline, thiomolybdic acid, and WTX101) is an investigational, oral, first-in-class copper protein-binding molecule being developed for the treatment of WD and is described in detail in International Publication No. WO2019 / 110619, the entirety of which is incorporated herein by reference. BC-TTM has the following structure: [ka] There is a need in the art for improved drug delivery systems for the delivery of BC-TTM for use in patient populations with different dosage needs. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. 2019 / 110619 Summary of the Invention [Means for solving the problem]

[0006] One embodiment of the present disclosure provides a minitablet formulation comprising bis-choline thiomolybdate in an amount ranging from about 1.00 mg to about 1.50 mg.

[0007] Another aspect of the present disclosure is a method for producing a method for manufacturing a semiconductor device comprising the steps of: Bis-choline tetrathiomolybdate in an amount of about 1.25 mg; Approximately 25% (by weight based on the weight of the minitablet core) of buffer; Approximately 66% (by weight based on the weight of the minitablet core) of a filler component; and about 0.75% (by weight based on the weight of the minitablet core) of a lubricant component.

[0008] Another aspect of the present disclosure provides a unit dose comprising one or more of the minitablets of the present disclosure. In certain embodiments, the unit dose of the present disclosure comprises two or more of the minitablets of the present disclosure.

[0009] Another aspect of the present disclosure provides a capsule, sachet, or stick pack comprising a unit dose of the present disclosure as described herein.The present disclosure provides a unit dose dispensing container configured to dispense a unit dose of the present disclosure as described herein.

[0010] Another aspect of the present disclosure provides a method for treating a copper metabolism-related disease or disorder in a subject. Such a method comprises administering to the subject one or more minitablets of the present disclosure as described herein or a unit dose of the present disclosure as described herein. In certain embodiments, the unit dose of the present disclosure can be provided in a unit dose container such as a capsule, sachet, stick pack, or can be dispensed from a unit dose dispenser as described herein.

[0011] Another aspect of the present disclosure provides a use for the manufacture of a pharmaceutical product of one or more of the minitablets as described herein or the unit doses of the present disclosure as described herein.In certain embodiments, the unit doses can be provided in unit dose containers such as capsules, sachets, stick packs, etc., or can be dispensed from unit dose dispensers as described herein.In certain embodiments, the use is for the manufacture of a pharmaceutical product for treating a copper metabolism-related disease or disorder in a subject.

[0012] These and other features and advantages of the claimed invention will become more fully understood from the following detailed description taken in conjunction with the appended claims, with the understanding that the scope of the claims is defined by the recitations therein, and not by the specific discussion of the features and advantages set forth in the description of the invention.

[0013] The accompanying drawings are included to provide a further understanding of the formulations and methods of the present disclosure, and are incorporated in and constitute a part of this specification. The drawings illustrate the description in conjunction with one or more embodiments of the present disclosure and serve to explain the principles and operation of the present disclosure. [Brief description of the drawings]

[0014] [Figure 1] Figure 1 shows the stability of a minitablet formulation of the present disclosure (F2G2; circles, solid line) and a comparative formulation (5 mg; triangles, dashed line) after 4 weeks of storage. The top chart shows the concentration (%) of total impurities in the formulation over time; the bottom chart shows the concentration (%) of BC-TTM in the formulation over time. [Diagram 2] Figure 1 shows the stability of minitablet formulations of the present disclosure, F2G2 (circles, solid line) and F1G2 (squares, dotted line) after 4 weeks of storage. The top chart shows the concentration (%) of total impurities in the formulation over time; the bottom chart shows the concentration (%) of BC-TTM in the formulation over time. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015] Before describing the disclosed processes and materials, it is to be understood that the aspects described herein are not limited to particular embodiments, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only, and is not intended to be limiting, unless specifically defined herein.

[0016] In view of the present disclosure, the methods and formulations described herein can be adapted by one of skill in the art to meet desired needs. The present disclosure provides improvements in treating copper metabolism-related diseases or disorders.

[0017] Wilson's disease (also called hepatic lens dysfunction) is an inherited disorder of copper transport. Wilson's disease is caused by various genetic mutations in the Cu-loading enzyme ATP7B (in humans). ATP7B facilitates the movement of Cu into the CP and its excretion via the bile canaliculi. The resulting defects in the hepatic excretion pathway lead to the accumulation of copper in tissues such as the liver, kidney, central nervous system / brain, and nuclear membrane, and copper levels remain elevated without treatment. Specifically, copper accumulation exceeds the capacity of the CP, resulting in free non-ceruloplasmin-bound copper ("NCC"), which circulates in the blood and accumulates in tissues and organs. This NCC can be loosely bound to plasma proteins (e.g., albumin, transcuprin, and low molecular weight peptides or amino acids) to form complexes ("labile bound copper" or "LBC").

[0018] In certain embodiments of the methods and uses of the present disclosure as described herein, the copper metabolism related disease or disorder is Wilson's disease.

[0019] In certain embodiments, the copper metabolism-related disease or disorder is copper poisoning (e.g., from high exposure to copper sulfate pesticides, ingesting copper-rich drinking water, overuse of copper supplements, etc.) In certain embodiments, the copper metabolism-related disease or disorder is copper deficiency, Menkes disease, or ceruloplasmin deficiency. In certain embodiments, the copper metabolism related disease or disorder is academic failure, acne, attention deficit / hyperactivity disorder, amyotrophic lateral sclerosis (ALS), atherosclerosis, autism, Alzheimer's disease, Candida overgrowth, chronic fatigue, liver cirrhosis, depression, elevated adrenergic activity, elevated copper protein, elevated norepinephrine activity, emotional lability, fibromyalgia, frequent anger, age-related impaired copper excretion, high anxiety, hair loss, liver disease, hyperactivity, hypothyroidism, estrogen intolerance, oral contraceptive intolerance, Kayser-Fleischer rings, learning disabilities, low dopamine activity, multiple sclerosis, neurological problems, oxidative stress, Parkinson's disease, poor concentration, poor concentration focus), weakened immune system, tinnitus, allergies, sensitivity to food dyes, sensitivity to shellfish, skin metal intolerance, skin sensitivities, sleep problems, and white spots on fingernails.

[0020] Advantageously, low dose formulations such as minitablets comprising bis-choline tetrathiomolybdate (BC-TTM) are provided that can be administered in various doses to a patient population with a continuing need for monitoring and dose adjustment throughout the patient's life. In particular, a patient's dosage can remain constant or can be adjusted to maintain therapeutic levels of BC-TTM and satisfactory copper levels. In some embodiments, the present disclosure further provides a capsule, sachet, or stick pack comprising one or more of the minitablets that allows administration of a specific dose of BC-TTM based on the patient's needs. In some other embodiments, the present disclosure further provides a unit dose dispensing container configured to dispense a unit dose of the minitablets.

[0021] In some embodiments, the minitablet formulations disclosed herein include BC-TTM in an amount of about 1.00 mg to about 1.50 mg. For example, BC-TTM may be present in an amount ranging from about 1.10 mg to about 1.40 mg, or from about 1.15 mg to about 1.35 mg, or from about 1.20 mg to about 1.30 mg, or from about 1.22 mg to about 1.28 mg, or from about 1.23 mg to about 1.27 mg, or from about 1.24 mg to about 1.26 mg. In some embodiments, the amount is In some embodiments, the minitablet formulations disclosed herein contain BC-TTM in an amount of about 1.25 mg.

[0022] In some embodiments, the minitablet formulations disclosed herein contain about 5% to about 10% (by weight based on the weight of the minitablet core, i.e., the weight of the tablet excluding the coating) of BC-TTM. In some embodiments, the minitablet formulations contain about 5%, about 5.5%, about 6.0%, about 6.5%, about 7.0%, about 7.5%, about 8.0%, about 8.5%, about 9.0%, about 9.5%, or about 10% (by weight based on the weight of the minitablet core) of BC-TTM. In certain embodiments, the minitablet formulations contain about 8.33% (by weight based on the weight of the minitablet core) of BC-TTM.

[0023] In some embodiments, the minitablet formulations disclosed herein include one or more buffers. As used herein, "buffer" refers to an excipient for maintaining the pH of the formulation. In certain embodiments, the buffer is sodium bicarbonate (NaHCO3). Sodium bicarbonate provides excellent stabilization of BC-TMM, advantageously allowing for the formulation of BC-TMM without the need for disintegrants for stabilization.

[0024] In some embodiments, the minitablet formulation comprises about 20% to about 30% (by weight based on the weight of the minitablet core) of buffer. For example, the buffer may be present in the range of about 22% to about 28% by weight, or about 23% to about 27% by weight, or about 24% to about 26% by weight, or about 20% to about 25% by weight, or about 25% to about 30% by weight based on the weight of the minitablet core. In some embodiments, the minitablet formulation comprises about 20% by weight, about 21% by weight, about 22% by weight, about 23% by weight, about 24% by weight, about 25% by weight, about 26% by weight, about 27% by weight, about 28% by weight, about 29% by weight, or about 30% by weight of buffer based on the weight of the minitablet core. In certain embodiments, the minitablet formulation comprises about 25% by weight of buffer based on the weight of the minitablet core.

[0025] In some embodiments, the minitablet formulation comprises BC-TTM and sodium bicarbonate present in a weight ratio ranging from about 10:90 to 40:60 (e.g., from about 20:80 to 30:70). In some embodiments, the minitablet formulation comprises BC-TTM and sodium bicarbonate in a ratio of about 10:90, about 20:80, about 25:75, about 30:70, or about 40:60. In some embodiments, the minitablet formulation comprises BC-TTM and sodium bicarbonate in a weight ratio of about 25:75.

[0026] In some embodiments, the minitablet formulations disclosed herein include a filler component. In certain embodiments, the filler component is tricalcium phosphate, dicalcium phosphate, lactose monohydrate, anhydrous lactose, spray-dried lactose, microcrystalline cellulose, powdered cellulose, silicified microcrystalline cellulose, starch, pregelatinized starch, or a combination thereof. In certain embodiments, the filler component is microcrystalline cellulose. In some embodiments, the minitablet formulations include about 60% to about 70% (by weight based on the weight of the minitablet core) of the filler component. For example, the filler component may be present in a range of about 62% to about 70% by weight, or about 63% to about 69% by weight, or about 64% to about 68% by weight, or about 65% to about 67% by weight based on the weight of the minitablet core. In some embodiments, the minitablet formulation comprises about 60%, about 61%, about 62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%, about 69%, or about 70% by weight of the filler component based on the weight of the minitablet core. In certain embodiments, the minitablet formulation comprises about 65% by weight of the filler component based on the weight of the minitablet core. In certain embodiments, the minitablet formulation comprises about 66% by weight of the filler component based on the weight of the minitablet core.

[0027] In some embodiments, the minitablet formulation disclosed herein comprises a lubricant component. In certain embodiments, the lubricant component is sodium stearyl fumarate, glyceryl behenate (i.e., Compritol 888 ATO), glyceryl monostearate, stearic acid, magnesium stearate, calcium stearate, hydrogenated vegetable oil, polyethylene glycol (PEG) 4000-6000, sodium lauryl sulfate (SLS), or a combination thereof. In certain embodiments, the lubricant component is sodium stearyl fumarate (sodium (E)-4-octadecoxy-4-oxobut-2-enoate). In certain embodiments, the lubricant component is a hydrophilic lubricant. In some embodiments, the minitablet formulation comprises about 0.5% to about 1% (by weight based on the weight of the minitablet core) of the lubricant component. For example, the lubricant component may be present in the range of about 0.6% to about 0.9% by weight, or about 0.65% to about 0.85% by weight, or about 0.7% to about 0.8% by weight, or about 0.72% to about 0.78% by weight, or about 0.73% to about 0.77% by weight, based on the weight of the minitablet core. In some embodiments, the minitablet formulation comprises about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, or about 1.0% by weight of the lubricant component, based on the weight of the minitablet core. In certain embodiments, the minitablet formulation comprises about 0.75% by weight of the lubricant component, based on the weight of the minitablet core.

[0028] In some embodiments, the minitablets further comprise a coating on the outer surface of the formulation. For example, the coating may be on the outer surface of the minitablet core comprising bis-choline tetrathiomolybdate and, if present, a buffer, a filler component, and / or a lubricant component. In some embodiments, the coating may comprise a seal coating, a subcoating, an enteric coating, or a combination thereof. In some embodiments, the seal coating comprises a hydrophobic material, such as, for example, carnauba wax. In some embodiments, the subcoating comprises a hydrophilic material. In some embodiments, the enteric coating comprises a methacrylic acid copolymer. In some embodiments, the coating may comprise at least two layers (e.g., three layers). In some embodiments, the coating comprises powdered carnauba wax as a seal coating, Opadry 200 Clear 203A190001 as a subcoating, or Acrylic-EZE White as an enteric coating, or a combination thereof.

[0029] Surprisingly, the minitablet formulations of the present disclosure as described herein maintain a high level of purity after extended storage. For example, in certain embodiments, the minitablet formulations of the present disclosure as described herein contain less than about 0.7%, or less than about 0.6%, or less than about 3.5%, or less than about 3.25%, or less than about 3%, or less than about 2.75%, or less than about 2.5%, or in the range of about 2% to about 3% total impurities after 4 weeks of storage at about 25° C. and about 60% relative humidity as determined by HPLC.

[0030] Common impurities observed in BC-TTM formulations are molybdenum impurities including, for example, TM0, TM1, TM2, and TM3. [ka] Other common impurities include polymeric molybdenum impurities such as dimer S6 and dimer S7 shown below. [ka]

[0031] In certain embodiments, the minitablet formulations of the present disclosure as described herein contain less than about 2%, or less than about 1.8%, or less than about 1.7%, or less than about 1.6%, or in the range of about 1% to about 2% total molybdenum impurities selected from one or more of TM0, TM1, TM2, and TM3 at about 25° C. and about 60% relative humidity as determined by HPLC for 4 weeks storage.

[0032] In certain embodiments, the minitablet formulations of the present disclosure as described herein contain less than about 0.7%, or less than about 0.6%, or less than about 0.5%, or less than about 0.4%, or less than about 0.3%, or in the range of about 0.1% to about 0.5%, of polymeric molybdenum impurities upon storage for 4 weeks at about 25° C. and about 60% relative humidity as determined by HPLC.

[0033] In certain embodiments, the minitablet formulations of the present disclosure as described herein have low levels of TM3 impurities after extended storage. In certain embodiments, the minitablet formulations of the present disclosure contain less than about 1.3%, or less than about 1.2%, or less than about 1.1%, or less than about 1%, or in the range of about 0.8 to about 1%, of TM3 impurities after 4 weeks of storage at about 25° C. and about 60% relative humidity as determined by HPLC.

[0034] In certain embodiments, the minitablet formulations of the present disclosure as described herein have low levels of dimeric S6 impurity after extended storage. In certain embodiments, the minitablet formulations of the present disclosure contain less than about 0.3%, or less than about 0.2%, or less than about 0.1%, or in the range of about 0.08 to about 0.12% dimeric S6 impurity after 4 weeks of storage at about 25° C. and about 60% relative humidity as determined by HPLC.

[0035] In some embodiments, the present disclosure further provides a unit dose comprising one or more of the minitablets of the present disclosure. In certain embodiments, the unit dose of the present disclosure comprises two or more of the minitablets of the present disclosure.

[0036] In certain embodiments, one or more of the unit doses of the present disclosure may be provided in a unit dose container. Examples of suitable unit dose containers include, but are not limited to, capsules, sachets, stick packs, or unit dose dispensers. Thus, a unit dose container of the present disclosure may contain one unit dose of the present disclosure. Such containers may include capsules, sachets, or stick packs. A unit dose container of the present disclosure may also contain two or more of the unit doses of the present disclosure. Examples of such containers include dispensers.

[0037] In some embodiments, the unit dose container of the present disclosure is configured to dispense a unit dose (such as one unit dose) of mini-tablets. Such unit dose containers allow patient populations that are unable to swallow tablets and capsules, such as pediatric and geriatric populations, to access and administer a dose of mini-tablets without having to swallow a whole tablet or capsule. In some embodiments, the unit dose container is a capsule (such as a sprinkle capsule), sachet, or stick pack that can be opened by the patient. In some embodiments, the unit dose container is a mini-tablet dispensing container, such as those marketed by Phillips Medisize.

[0038] In some embodiments, the unit dose comprises about 2.5 mg, about 3.75 mg, about 5 mg, about 6.25 mg, about 7.5 mg, about 8.75 mg, about 10 mg, about 11.25 mg, about 12.5 mg, about 13.75 mg, about 15 mg, about 20 mg, or about 30 mg of BC-TTM. In some embodiments, the unit dose comprises about 2.5 mg, about 3.75 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, or about 12 mg of BC-TTM. In some embodiments, the unit dose comprises about 15 mg, about 20 mg, about 25 mg, or 30 mg of BC-TTM. In some embodiments, the unit dose comprises about 15 mg of BC-TTM.

[0039] In some embodiments, a unit dose container, such as a reclosable capsule, sachet, stick pack, etc., provides a dose of BC-TTM of about 5 mg to about 30 mg. In some embodiments, a unit dose container provides a dose of BC-TTM of about 2.5 mg to about 12.5 mg, e.g., about 2.5 mg, or about 5 mg, or about 10 mg. In some embodiments, a unit dose container provides a dose of BC-TTM of about 15 mg to about 30 mg, e.g., about 15 mg, or about 20 mg, or about 30 mg.

[0040] In some embodiments, the unit dose container contains at least about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24 1.25 mg minitablets. In some embodiments, the unit dose container contains 6 1.25 mg minitablets. In some embodiments, the unit dose container contains more than 24 1.25 mg minitablets.

[0041] In some embodiments, the unit dose container is a minitablet dispenser configured to dispense a unit dose of minitablets containing about 2.5 mg, 3.75 mg, about 5 mg, about 6.25 mg, about 7.5 mg, about 8.75 mg, about 10 mg, about 11.25 mg, about 12.5 mg, about 13.75 mg, about 15 mg, about 20 mg, or about 30 mg of BC-TTM. In some embodiments, the dispenser is configured to dispense a unit dose of minitablets containing about 2.5 mg, about 3.75 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, or about 12 mg of BC-TTM. In some embodiments, the dispenser is configured to dispense a unit dose of minitablets containing about 15 mg, about 20 mg, about 25 mg, or about 30 mg of BC-TTM. In some embodiments, the dispenser is configured to dispense a unit dose of about 15 mg minitablets of BC-TTM.

[0042] In some embodiments, the dispenser dispenses minitablets providing a unit dose of about 5 mg to about 30 mg of BC-TTM. In some embodiments, the dispenser dispenses minitablets providing a unit dose of about 2.5 mg to about 12.5 mg, e.g., about 2.5 mg, or about 5 mg, or about 10 mg of BC-TTM. In some embodiments, the dispenser dispenses minitablets providing a unit dose of about 15 mg to about 30 mg, e.g., about 15 mg, or about 20 mg, or about 30 mg of BC-TTM.

[0043] In some embodiments, the dispenser dispenses a unit dose of minitablets that includes at least about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24 1.25 mg minitablets. In some embodiments, the dispenser dispenses a unit dose of 6 1.25 mg minitablets. In some embodiments, the dispenser dispenses a unit dose of minitablets that includes more than 24 1.25 mg minitablets.

[0044] The dispenser, in certain embodiments, is configured to dispense unit doses of the present disclosure, conveniently containing two or more unit doses, such as 15, or 30, or 60 unit doses. Thus, in certain embodiments, the dispenser contains at least about 30 to about 720 1.25 mg minitablets (e.g., a 30 day supply). In certain embodiments, the dispenser contains at least about 90 to about 360 1.25 mg minitablets. In certain embodiments, the dispenser contains about 90, or 18, or about 360 1.25 mg minitablets.

[0045] As mentioned above, the unit dose container of the present disclosure provides a convenient means for providing a dose of mini-tablets. For example, the capsule, sachet, or stick pack is configured to be opened by the patient (e.g., sprinkle capsules, etc.). Thus, in some embodiments of the method of the present disclosure, administration includes opening the capsule, sachet, or stick pack or dispensing a unit dose of mini-tablets from a mini-tablet dispenser and providing the contents of the mini-tablets to a food (such as a soft acidic food). Without being bound by theory, it is believed that the mini-tablets may be administered with an acidic soft food to protect the enteric coating until the site of absorption in the gastrointestinal tract. In some embodiments, one or more mini-tablets are administered by sprinkling one or more mini-tablets on a soft acidic food such as applesauce or yogurt. In some embodiments, administering one or more of the mini-tablets with food provides a statistically equivalent average bioavailability to one or more of the mini-tablets administered without food.

[0046] One or more minitablets or unit doses of the present disclosure as described herein can be administered daily in the method and use of the present disclosure as described herein.For example, in certain embodiments, one or more minitablets or unit doses are administered once a day.In certain embodiments of the method and use of the present disclosure as described herein, one or more minitablets or unit doses can be administered every other day.

[0047] In certain embodiments of the methods and uses of the present disclosure as described herein, the dose comprises BC-TMM in an amount of about 15 mg. For example, in certain embodiments, the dose comprises a unit dose comprising a plurality of minitablets or a plurality of minitablets having a combined amount of 15 mg of BC-TMM.

[0048] In certain embodiments of the methods and uses of the present disclosure as described herein, one or more minitablets or unit doses are administered in fasting state.For example, in certain embodiments, fasting state is the state after overnight fasting.In certain embodiments, administration is performed in fasting state, for example, at least 1 hour before a meal or at least 2 hours after a meal.

[0049] The methods of the present disclosure are further illustrated by the following examples, which are not to be construed as limiting the disclosure in scope or spirit to the specific procedures and compounds described therein. EXAMPLES

[0050] Example 1: Preparation of low dose formulations ("minitablets") Various low dose formulations of BC-TTM, referred to herein as "minitablets", containing 1.25 mg of BC-TTM and excipients were prepared as shown in Tables 1 and 2. Upon final blending of BC-TTM with excipients, tablet cores were produced using a compression machine according to the method commonly used for the manufacture of tablet dosage forms. The tablet cores were then subjected to coating according to a common coating method. Formulation #1, generations 1, 2 and 3 contained the hydrophobic lubricant magnesium stearate (Table 1). For formulation #2, generations 1, 2, 3 and 4, the lubricant was changed to the hydrophilic lubricant sodium stearyl fumarate (Table 2).

[0051] [Table 1]

[0052] [Table 2]

[0053] Example 2: 4-week accelerated stability of minitablets The objective of the stability study was to evaluate the stability profile of several BC-TTM minitablet formulations. Stability was evaluated using observation of one tablet (for product appearance) and HPLC / UV (200-400 nm) analysis of an injection from one tablet sample preparation (for assay of BC-TTM and impurity content). The stability of the minitablets was compared to a tablet containing 5 mg of ALX1840 with the formulation as shown in Table 3.

[0054] The stability of minitablets of the present disclosure was evaluated at the start ("ATST"), at 1 week ("1W"), at 2 weeks ("2W"), and at 4 weeks ("4W") when stored at 5°C, 60% relative humidity (RH) at 25°C, and 75% RH at 40°C. Table 4 provides an evaluation of Formulation #1, Generation 2 (F1G2), of Example 1, Table 5 provides an evaluation of Formulation #2, Generation #2 (F2G2), of Example 1, and Table 6 provides an evaluation of the 5 mg tablets. As used herein, LTLOQ means "below the lower limit of quantification" and as used herein, ND means "not determined." For Tables 4-7 and 10, the reported amount of TM0 is in its anionic form ([MoO4] 2- ) of TM0, whereas TM0 in the remainder of this disclosure is reported with respect to its choline salt form. Thus, the amount of "Total Impurities" reported in Tables 4-7 and 10 was calculated using the amount of TM0 in its anionic form, whereas the amount of "Total Impurities" reported in the remainder of this disclosure was calculated using the amount of TM0 in its choline salt form.

[0055] Surprisingly, the 1.25 mg F2G2 minitablets showed greater stability compared to the 5 mg tablets, as illustrated by the lower concentration (%) of total impurities over time and the higher concentration (%) of BC-TTM over time (Figure 1 and Table 7). In addition, Figure 2 illustrates that the 1.25 mg F2G2 minitablets also showed greater stability compared to the 1.25 mg F1G2 minitablets.

[0056] [Table 3]

[0057] [Table 4]

[0058] [Table 5]

[0059] [Table 6]

[0060] [Table 7]

[0061] Example 3: Preparation of minitablets The minitablets of the present disclosure were prepared at a manufacturing scale. The manufacturing recipe for the minitablets is provided in Table 8 below. Smaller or larger batches may be manufactured using this ingredient and ratio. The cores of the minitablets were prepared using a dry granulation process. Briefly, at the final blend, the minitablet cores were manufactured using a compression machine to match their desired physical attributes. The minitablet cores were then subjected to seal coating, subcoating, and final enteric coating. The minitablet manufacturing process used commercially available pharmaceutical processing equipment commonly used to manufacture tablet dosage forms.

[0062] [Table 8]

[0063] Description of manufacturing process and process control The manufacturing process consisted of blending the drug substance and excipients in a dry granulation process. The final blend was then compressed into minitablet cores. The coating process began with seal coating the cores with carnauba wax. The seal coated tablets were then subjected to subcoating using Opadry 200 Clear followed by enteric coating with Acryl-EZE White. The main processing steps were pre-roller compaction blending, ribbon roller compaction and milling, final blending of the drug substance granulation with extragranular excipients, minitablet compression, seal coating, subcoating and enteric coating.

[0064] Pre-Roller Compaction Blending: Prior to processing, it was confirmed for each batch that BC-TTM was dispensed within two days of the start of production. Sodium Bicarbonate, USP Grade 1 Powder-Increment 1 was filled into a 15L container, then BC-TTM was added to the same 15L container. The bag containing the remaining BC-TTM was rinsed with sodium bicarbonate, then USP Grade 1 Powder-Increment 2 was added to the same 15L container. These materials were then blended in the 15L container for 5 minutes at 10 RPM. Sodium Bicarbonate, USP Grade 1 Powder-Increment 3 was then filled into the 15L container, and the materials were blended for 10 minutes at 10 RPM.

[0065] The blended ingredients were then discharged into an intermediate container and then comminuted using a Quadro Comil equipped with a 032R screen (approximately 812 microns). The comminuted material was then returned to the same 15 L container and mixed for an additional 5 minutes at 10 RPM. Microcrystalline cellulose, NF (Avicel PH-112) was comminuted by passing through the same Comil equipped with a 032R screen and collected in a clean suitable container. The comminuted microcrystalline cellulose, NF (Avicel PH-112) was charged into the same 15 L container and mixed for 15 minutes at 10 RPM.

[0066] An equal volume of the blend from the 15 L bin was added to the Sodium Stearyl Fumarate, NF (intragranular) and mixed by inverting the bag for approximately 20 seconds. This mixture was simultaneously screened through a 20 mesh hand screen directly into the 15 L bin and blended at 10 RPM for 5 minutes. The pre-roller compacted blend was then discharged into an intermediate bin and the yield and accountability were calculated.

[0067] Roller Compaction and Milling of Ribbons: The pre-compacted blend was roller compacted using an Alexanderwerks WP120 roller compactor equipped with 40 mm top smooth / bottom angle rollers and a chiller set at 15° C. The ribbons were milled at 95 RPM using an integrated in-line mill on the Alexanderwerks WP120 roller compactor equipped with a 1.0 mm coarse screen and a 0.63 mm fine screen.

[0068] Ribbon and milled granulation samples were collected at the beginning, middle, and yield of roller compaction. Upon completion of roller compaction, the milled granulation was collected in an intermediate container for immediate further processing.

[0069] Final Blending: The weight of the extragranular component (Sodium Stearyl Fumarate, NF) was adjusted based on the yield of granulation recovered from the roller compaction and milling steps. Initially, approximately 50% of the milled granulation was filled into a 15L container. An equal volume of milled granulation from the remaining granulation was added to the Sodium Stearyl Fumarate, NF (extragranular) and mixed by inverting the bag for approximately 20 seconds, then hand screened through a 20 mesh screen directly into the container, and then the remaining milled granulation filled was filled into the container. The mixture was blended at 10 RPM for 5 minutes.

[0070] A homogenous sample of the final blend was collected from the container in triplicate at 10 locations using a disposable 0.5 ml sample thief. Approximately 100 g samples from the container were also collected and the final blend was then discharged into a foil bag that was double lined with a polyethylene bag with one desiccant in the headspace of the outer polyethylene bag. Air was removed from the polyethylene bag before closing with a zip tie. Similarly, air was removed from the foil bag and then purged with nitrogen for approximately 3 minutes before heat sealing. Yields and theoretical yields were calculated. The foil bag was then placed into a foil lined fiber drum and stored at 2-8°C.

[0071] Minitablet Cores: The final blend of BC-TTM (8.33% by weight based on the weight of the core) was compressed into minitablet cores using a Korsch XL 100 Pro Tablet Press equipped with 3 mm Round Multi Tip tooling and a first feeder. The compressed tablets were de-dusted using a Key tablet de-duster and metal tested using a Lock Met30+ metal detector. The minitablets were compressed to a target weight of 15 mg / unit and met other physical attributes. In-process samples were collected and tested for physical attributes at predetermined time intervals during compression to ensure product quality.

[0072] The bulk core tablets were collected in a foil bag that was double lined with a polyethylene bag with a desiccant in the headspace of the outer polyethylene bag. As much air as possible was removed from the polyethylene bag before closing with a zip tie. The foil bag was also processed to remove as much air as possible and then purged with nitrogen for approximately 3 minutes before heat sealing. The foil bag was then placed in a foil lined fiber drum and stored at 2-8°C.

[0073] Enteric Coating of Minitablets: Three sublots with approximately equal pan fill sizes and equivalent coating processes are required to coat the entire theoretical batch.

[0074] A seal coat coating of powdered carnauba wax, NF#1, was applied to the minitablet cores using a pan coating system. The core minitablets were seal coated to a theoretical weight gain of 1% in a Compu-Lab coater equipped with a 15" pan.

[0075] The subcoat coating dispersion was prepared at 20% solids using Opadry 200 Clear (203A190001) coating system and purified water. The subcoated tablets were subcoated in a Compu-Lab coater equipped with a 15" pan to a theoretical weight gain of 20% ± 1%.

[0076] The enteric coating dispersion was prepared at 20% solids using Acrylic-EZE White Coating System and purified water. The subcoated tablets were coated in a Compu-Lab coater equipped with a 15" pan to a theoretical weight gain of 35% ± 1%.

[0077] Upon completion of coating, the bulk enteric coated tablets were collected in a foil bag that was double lined with a polyethylene bag with one desiccant in the headspace of the outer polyethylene bag. As much air as possible was removed from the polyethylene bag before closing with a zip tie. The foil bag was also removed as much air as possible, and then it was purged with nitrogen for approximately 3 minutes before heat sealing. The foil bag was then placed back into the foil-lined fiber drum.

[0078] A summary of the in-process controls for pharmaceutical manufacturing is provided in Table 9.

[0079] [Table 9]

[0080] There were no significant differences between the 1.25 mg minitablet formulations manufactured in different batches (Table 10).

[0081] [Table 10]

[0082] Example 4: Six-Month Stability of Capsules Containing Low-Dose Formulations The minitablets prepared according to Example 3 were placed into hydroxypropyl methylcellulose (HPMC) sprinkle capsules. Each HPMC capsule contained four individual 1.25 mg minitablets. The capsules were stored in 60 cc HDPE WM round bottles ((0060Hl-01)(33 / 400)Q024847) closed with DPC CRH11100 33MM WHT SECURX RIBD SIDE PP CRC TXT (7821H1-G1 263131). Each bottle contained 30 capsules.

[0083] Capsule stability was evaluated based on product appearance, assay / impurities, dissolution, and moisture when stored at 5° C. and 25° C. / 60% RH. Stability data measured at 0, 1, 2, 3, 4, 5, 6, and 12 months for samples stored at 5° C. and 25° C. / 60% RH conditions, respectively, are provided in Tables 11 and 12. LTLOQ, as used herein, means "lower limit of quantification" and ND, as used herein, means "not determined."

[0084] [Table 11]

[0085] [Table 12]

[0086] Another set of capsules containing another batch of 1.25 mg minitablets (so-called "second batch") prepared according to Example 3 (4 minitablets per capsule prepared and stored as above except the bottles were closed with 33 mm SCRX RIBD SIDE WHT PP CRC TXT TOP (HS130-35 7903Hl-1Cl 263455)) was also tested for long term stability against the criteria provided in Table 14. Table 13 provides the results of the stability evaluation at 3 months storage at 5°C and 25°C / 60% RH, 6 months storage at 25°C / 60% RH, and 12 months storage at 5°C.

[0087] [Table 13]

[0088] [Table 14]

[0089] Example 5: Relative bioavailability of two oral formulations of BC-TTM in healthy adult participants A phase 1, randomized, two-period, two-sequence, crossover with parallel group expansion, open-label study was conducted to compare the relative bioavailability of two oral formulations of BC-TTM in healthy adult participants. The objective of the study was to evaluate the relative bioavailability of the 1.25 mg enteric coated (EC) minitablet formulation of BC-TTM compared to the 15 mg EC tablet of BC-TTM to evaluate dose proportionality between 2.5 mg (2 x 1.25 mg), 5 mg (4 x 1.25 mg), 10 mg (8 x 1.25 mg), 15 mg (12 x 1.25 mg), and 30 mg (24 x 1.25 mg) EC minitablet doses. The 15 mg EC tablet of BC-TTM used in the study had a formulation consisting of the ingredients listed in Table 3. The 1.25 mg EC minitablets of BC-TTM were prepared according to the pharmaceutical formulation formulation in Example 3 and Table 8.

[0090] This was a two-period, two-sequence, parallel extension, crossover study designed to evaluate the relative bioavailability of equal doses of BC-TTM administered as a single 15 mg EC tablet versus 1.25 mg EC minitablets, and to evaluate dose proportionality between 2.5 mg (2 × 1.25 mg), 5 mg (4 × 1.25 mg), 10 mg (8 × 1.25 mg), 15 mg (12 × 1.25 mg), and 30 mg (24 × 1.25 mg) EC minitablet doses in the dose proportionality extension period. The stability and tolerability of the two formulations of BC-TTM in healthy participants were also evaluated. The pharmacokinetics (PK) of BC-TTM in plasma as measured via total molybdenum (Mo) and plasma ultrafiltrate (PUF) Mo were determined.

[0091] [Table 15]

[0092] [Table 16]

[0093] [Table 17]

[0094] The study had a screening period (day -28 to day -2), a two-way crossover period consisting of two treatment periods (days 1 to 11, respectively), and a dose-proportional extension period. After completing the screening period, enrolled participants were admitted to the Clinical Research Unit (CRU) on day -1 for dosing on day 1 during treatment period 1. If discharged after treatment period 1, participants were readmitted to the CRU during treatment period 2 after a minimum of 14 days washout from the previous dose, and also for the dose-proportional extension period after a minimum of 14 days washout. An end-of-study (EOS) visit was conducted 14 days (± 2 days) after dosing of BC-TTM in the dose-proportional extension period.

[0095] The two-way crossover period was a randomized, open-label, two-way (two periods, two sequences), crossover design to evaluate the relative bioavailability of 12 × 1.25 g EC minitablets compared with the 15 mg EC tablets currently used in clinical trials. Participants were randomly assigned to one of two treatment sequences. Randomized treatment assignment was based on baseline body mass index (BMI). Two strata for BMI (<25, 25 to <32 kg / m2) were included. 2 ) was used: Treatment A: 12 × 1.25 mg EC minitablets of BC-TTM Treatment B: A single 15 mg EC tablet of BC-TTM (reference tablet, currently being studied in the Phase 3 study WTX101-301)

[0096] [Table 18]

[0097] Blood samples for PK analysis of total Mo and PUF Mo (as surrogate measures of BC-TTM PK) and pharmacodynamics (PD) / biomarkers were collected pre-dose on Day 1, and at 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours post-dose (Day 2), then at 24-hour intervals on Days 3, 4, 5, 6, 7, 8, 9, 10, and 11 during each dosing period.

[0098] The 336-hour sample for dosing period 1 was collected prior to dosing of dosing period 2. Participants could be discharged on Day 11 of each dosing period following completion of all treatments and review of all stability data. End of dosing period 2 occurred on Day 15±2 of dosing period 2, and a 336-hour PK sample for dosing period 2 was collected.

[0099] The dose proportionality extension period was a re-randomized, open-label, parallel-group, design to evaluate dose proportionality between the 2.5 mg (2 × 1.25 mg), 5 mg (4 × 1.25 mg), 10 mg (8 × 1.25 mg), and 30 mg (24 × 1.25 mg) EC minitablet doses. The 15 mg (12 × 1.25 mg) dose was not repeated during the dose proportionality extension period.

[0100] The dose-proportional extension period occurred after the completion of the two-way crossover period of the study and a washout period of at least 14 days. Participants were re-randomized as follows: Treatment C (N = 10–12): BC-TTM 2.5 mg (2 × 1.25 mg EC minitablets) Treatment D (N = 10–12): BC-TTM 5 mg (4 × 1.25 mg EC minitablets) Treatment E (N = 10–12): BC-TTM 10 mg (8 × 1.25 mg EC minitablets) Treatment F (N = 10–12): BC-TTM 30 mg (24 × 1.25 mg EC minitablets)

[0101] Dose proportionality assessment included data obtained from Treatment A in the two-way crossover period (12 x 1.25 mg EC minitablets) representing a dose of 15 mg.

[0102] Re-randomization treatment assignment was based on baseline body mass index (BMI). There were two strata for baseline BMI: <25 and 25–<32 kg / m 2 ) was used. Participants were randomly assigned equally to each treatment using block randomization.

[0103] Participants could be discharged on Day 11 of the dose-proportional extension period after completion of all treatments and review of all stability data.

[0104] Participants may be asked or required to remain in the CRU during the two-way crossover period and / or at the end of the dose proportional extension period prior to the End of Study (EOS) visit, for their own safety and to maintain the integrity of the conduct of the study.

[0105] Final data showed that of 48 randomized participants, 44 participants completed the two-way crossover period and 40 participants completed the dose-proportional extension period. All 48 (100%) participants randomized to the two-way crossover period were included in the safety, PKDS-CO, and full analysis sets, and all 41 (100%) participants randomized to the dose-proportional extension period were included in the safety, PKDS-E, and full analysis sets.

[0106] The PK profile of BC-TTM and plasma total molybdenum and PUF molybdenum as surrogate measures of PK parameters were comparable between single doses of BC-TTM administered as 12 x 1.25 mg EC minitablets (total dose of 15 mg) and as 1 x 15 mg EC tablet. max With 90% CI for total molybdenum, there were no clinically relevant differences between the two treatment formulations under fasting conditions in healthy participants, and the AUC t and AUC ∞ fell within the bioequivalence limits of 80% to 125%.

[0107] [Table 19]

[0108] Plasma total molybdenum PK parameters generally showed a dose-proportional increase from 2.5 mg to 30 mg for the EC minitablet formulation of BC-TTM. Plasma PUF molybdenum PK parameters showed a less than dose-proportional increase from 2.5 mg to 30 mg for the EC minitablet formulation of BC-TTM. The PK of BC-TTM was not clearly affected by weight or BMI.

[0109] [Table 20]

[0110] Plasma total molybdenum C normalized to dose max and AUC t The values ​​moderately decreased with increasing dose, and the AUC ∞ For PUF molybdenum, the dose-normalized plasma exposure values ​​decreased with increasing dose, suggesting that PUF molybdenum exposure for BC-TTM increased in a non-dose-proportional manner within the dose range of 2.5 mg to 30 mg of BC-TTM for the EC minitablet formulation (Table 18).

[0111] [Table 21]

[0112] Results of the analysis of potential formulation differences between treatments A and B suggest that there were no clinically meaningful differences in the PK parameters of BC-TTM between the two treatments or formulations. Plasma total molybdenum (C max , AUC t , and AUC ∞ ) and PUF molybdenum (C max The geometric mean ratio (90% CI) of the PUF molybdenum AUC with a geometric mean ratio (90% CI) of 101.2% (70.6% to 145.1%) was entirely within the 90% CI bounds of the default invalidation of 80% to 125%. twas excluded, as its lower and upper boundaries extend slightly outside the 80% to 125% invalidation boundaries (Table 19).

[0113] [Table 22]

[0114] Total molybdenum: For the dose range of 2.5 mg to 15 mg and the dose range of 2.5 mg to 30 mg, max and AUC t The dose proportionality criterion for was met with 90% CI slopes falling inside the critical interval defined as ([1 + ln(0.5) / ln(ρ), 1 + ln(2) / ln(ρ)]). However, for the dose ranges of 2.5 mg to 5 mg and 2.5 mg to 10 mg, C max and AUC t The dose proportionality criteria for AUC ∞ For , the dose proportionality criterion was met only for the dose range of 2.5 mg to 10 mg, but not for the dose ranges of 2.5 mg to 15 mg and 2.5 mg to 30 mg. Overall, the power model-based dose proportionality analysis results demonstrate that the increase in total molybdenum exposure was roughly dose proportional across the doses examined, from 2.5 mg to 30 mg.

[0115] PUF Molybdenum: C max and AUC t Dose proportionality criteria for values ​​were not met for any dose range. Power model-based dose proportionality analysis results showed that increases in PUF molybdenum exposure did not meet dose proportionality over the examined dose range of 2.5 mg to 30 mg, most likely due to the very high variability of PUF molybdenum concentrations versus plasma total molybdenum.

[0116] [Table 23]

[0117] There were no obvious differences in BC-TTM PD parameters (plasma total copper and PUF copper concentrations) between the 12 × 1.25 mg EC minitablets and the 15 mg reference EC tablet. Maximum plasma total copper concentrations occurred 8 hours post-dose, then gradually decreased, eventually returning to pre-dose baseline concentrations by 96-120 hours post-dose. Pre-dose baseline mean plasma total copper concentrations for Treatments A and B were 988 and 986 ng / mL, respectively, and transiently increased to mean maximum values ​​of 1230 and 1210 ng / mL, respectively, at 8 hours post-dose.

[0118] Beginning 8 hours after dosing, plasma total copper concentrations gradually decreased, with mean concentrations falling to <11% above predose baseline at 48 hours after dosing. By 96-120 hours after dosing, total copper concentrations had returned to predose baseline concentrations. PUF copper concentrations were much lower than total copper concentrations (mean values ​​<10 ng / mL) at all sampling time points, limiting the opportunity for quantitative assessment.

[0119] Summary statistics were calculated for absolute change and percent change from baseline plasma total copper concentrations after treatments C, D, E, and F. Treatment A from the two-way crossover period was also included in the comparison. For treatment C (2.5 mg BC-TTM, the lowest BC-TTM dose), the plasma total concentration versus time profiles remained stable overall. The plasma total copper concentration versus time profiles after treatments D, E, and F showed similar trends to the profiles of treatments A and B. Plasma total copper concentrations reached a maximum 6-12 hours post-dose and were centered at 8 hours, with maximum mean percent change (increase) from baseline (0.5 hours pre-dose) for treatments C, D, E, A, and F being approximately 2%, 10%, 18%, 26%, and 31%, respectively. The maximum mean percent change was dose-dependent but did not meet dose proportionality.

[0120] After the 12-h postdose time point, plasma total copper concentrations gradually decreased, reaching median changes from baseline within approximately <15% of predose baseline at 48 h postdose. Total copper concentrations had returned to predose baseline levels by 120–144 h postdose. PUF copper concentrations were much lower than total copper concentrations (mean values ​​<10 ng / mL) at all sampling time points, limiting the opportunity for quantitative assessment.

[0121] BC-TTM had an acceptable safety profile and was generally well tolerated in healthy adult participants when administered in single oral doses of 2.5 mg to 30 mg as EC minitablets and as 15 mg EC tablets, with no notable differences in the incidence of TEAEs. No deaths or TESAEs were reported. All TEAEs were grade 1 or 2 in severity, except for two events of increased blood creatine phosphokinase blood levels of grade 4 severity reported in two (4.3%) participants after treatment B during the two-way crossover period. The incidence of TEAEs was similar between treatment A (12 × 1.25 mg EC minitablets of BC-TTM) and treatment B (single 15 mg EC reference tablet of BC-TTM), and no dose correlation was observed in treatments C–F (2.5 mg to 30 mg BC-TTM administered as 1.25 mg EC minitablets).

[0122] Example 6. Food vehicle testing Food testing was performed to observe and test the integrity and stability of 1.25 mg minitablets of BC-TTM once introduced into a food vehicle. 1.25 mg minitablets of BC-TTM were prepared according to the pharmaceutical manufacturing recipe in Example 3 and Table 8. The minitablets were tested at 5 mg (4 x 1.25 mg) and 1.25 mg doses in either yogurt or applesauce. Samples were immersed in the food vehicle at the allotted time points at both room temperature and 5°C food storage conditions. Samples were then removed from the food vehicle and visually observed and tested.

[0123] Specifically, the tests were performed as follows: 1. Samples were tested in triplicate. 2. Both room temperature and 5°C storage conditions of the food vehicle were tested to determine if storage of the food vehicle affected the integrity of the samples. 3. Samples were tested at 5 mg doses (4 x 1.25 mg) and at 1.25 mg doses. 4. Applesauce soaking times: 5, 7.5, 10, 12.5, and 15 minutes. 5. Yogurt soaking times: 5, 10, 15, 30, 45, 60, 90, and 120 minutes. 6. Delivery technique: Minitablets were placed on top of the food vehicle and stirred from top to bottom a total of three times to best represent handling that may occur during administration. 7. No food vehicle was added to the dissolution vessel after sample immersion.

[0124] yogurt Yoplait Original French Vanilla Low Fat Yogurt (6 oz.) (pH 4.24) was the brand used for the yogurt food vehicle.

[0125] A 5 mg dose (4 x 1.25 mg) or a 1.25 mg dose was placed on top of the yogurt and the mini-tablets were stirred with a spoon from bottom to top a total of three times to ensure the sample was completely covered. The spoon was then removed and a foil lid was placed on top to cover. The mini-tablets were immersed in the yogurt for the following time points: 5, 10, 15, 30, 45, 60, 90, and 120 minutes. Samples from each time point were tested in both 5°C and room temperature food vehicle storage conditions, n=3. Room temperature samples were left on the lab countertop for the duration of the food immersion, while 5°C samples were immediately placed in 5°C storage after introduction into the yogurt. After the assigned time points, the samples were removed from the yogurt and observed.

[0126] For the 5 mg dose, minitablets were placed in the basket of dissolution apparatus 1 and transferred to an acid stage bath (500 mL, 0.1 N HCl, 37° C.±0.5° C.) set at a rotation speed of 100 rpm for 2 hours. Samples were then removed from the acid bath for observation and transferred to a buffer stage bath (500 mL, modified artificial intestinal fluid pH 7.5±0.05, 37° C.±0.5° C.) set at a rotation speed of 75 rpm. Samples were taken at 10, 12.5, 15, 20, and 30 minutes. After the 20 minute sampling time point, the rotation speed was increased to 250 rpm. Samples were then analyzed using HPLC.

[0127] For the 1.25 mg dose, the minitablets were placed in the minivessel acid stage bath (75 mL, 0.1 N HCl 37° C.±0.5° C.) of the Dissolution Apparatus 2 set at a rotation speed of 100 rpm for 2 hours. After the 2 hour acid stage, the minitablets were observed and buffer solution (25 mL, 0.25 M sodium phosphate tribasic, preheated to 37° C.±0.5° C.) was added to the vessel. The paddle rotation speed was reduced to 75 rpm and samples were taken at 10, 12.5, 15, 20, and 30 minutes. After the 20 minute sampling time point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC.

[0128] For the 5 mg dose (4 x 1.25 mg), the samples tested in yogurt showed no visible signs of swelling or discoloration throughout the test. The integrity of the minitablets was not compromised by introduction into yogurt.

[0129] For the 1.25 mg dose, samples tested in yogurt showed no visible signs of swelling or discoloration throughout the study. The integrity of the minitablet coating was not compromised by introduction into yogurt.

[0130] Applesauce Mott's Applesauce (4 oz.), pH 3.68, was the brand used for the applesauce food vehicle.

[0131] A 5 mg dose (4 x 1.25 mg) or a 1.25 mg dose was placed on top of the applesauce and the mini-tablets were stirred with a spoon from bottom to top a total of three times to ensure the sample was completely covered. The spoon was then removed and a foil lid was placed on top to cover. The mini-tablets were immersed in the applesauce for the following time points: 5, 7.5, 10, 12.5, and 15 minutes. Samples from each time point were tested in both 5°C and room temperature food vehicle storage conditions, n=3. Room temperature samples were left on the lab countertop for the duration of the food immersion, while 5°C samples were immediately placed in 5°C storage after introduction into the applesauce. After the assigned time points, the samples were removed from the food vehicle and observed.

[0132] For the 5 mg dose, minitablets were placed in the basket of dissolution apparatus 1 and transferred to an acid stage bath (500 mL, 0.1 N HCl, 37° C.±0.5° C.) set at a rotation speed of 100 rpm for 2 hours. Samples were then removed from the acid bath for observation and transferred to a buffer stage bath (500 mL, modified artificial intestinal fluid pH 7.5±0.05, 37° C.±0.5° C.) set at a rotation speed of 75 rpm. Samples were taken at 10, 12.5, 15, 20, and 30 minutes. After the 20 minute sampling time point, the rotation speed was increased to 250 rpm. Samples were then analyzed using HPLC.

[0133] For the 1.25 mg dose, the minitablets were placed in the minivessel acid stage bath (75 mL, 0.1 N HCl 37° C.±0.5° C.) of the Dissolution Apparatus 2 set at a rotation speed of 100 rpm for 2 hours. After the 2 hour acid stage, the minitablets were observed and buffer solution (25 mL, 0.25 M sodium phosphate tribasic, preheated to 37° C.±0.5° C.) was added to the vessel. The paddle rotation speed was reduced to 75 rpm and samples were taken at 10, 12.5, 15, 20, and 30 minutes. After the 20 minute sampling time point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC.

[0134] For the 5 mg dose, the minitablets were observed after immersion in applesauce with no visible signs of discoloration or decomposition. All samples were then transferred to a 2 hour acid stage bath. After the 2 hour acid stage, the minitablets were observed. All 5° C. time point samples remained intact and showed no signs of swelling or discoloration. The 5 minute and 7.5 minute room temperature time point samples also remained intact and showed no visible signs of discoloration or decomposition. All room temperature time point samples after 7.5 minutes (10, 12.5, and 15 minutes) had decomposed in the acid stage. Only the 5° C. samples and the 5 minute and 7.5 minute room temperature time point samples were able to continue into the buffer stage.

[0135] For the 1.25 mg dose, the minitablets were observed after immersion in applesauce with no visible signs of discoloration or decomposition. All samples were then transferred to a 2 hour acid stage bath. After the 2 hour acid stage, the minitablets were observed. All 5° C. time point samples remained intact and showed no signs of swelling or discoloration. The room temperature 5 minute and 7.5 minute time point samples also remained intact and showed no visible signs of swelling or decomposition. All room temperature time point samples after 7.5 minutes (10, 12.5, and 15 minutes) showed slight signs of swelling throughout the acid stage but no visible signs of discoloration or decomposition.

[0136] The results summarized in this example confirm that 1.25 mg enteric coated minitablets of BC-TTM are stable after incorporation into food vehicles. In applesauce, the minitablets are stable for up to 7.5 minutes at room temperature for the 5 mg dose (4 x 1.25 mg) and for up to 15 minutes at 5°C (refrigerated), and for the 1.25 mg dose, for both room temperature and 5°C (refrigerated) storage conditions. In yogurt, the minitablets are stable for up to 120 minutes at both room temperature and 5°C (refrigerated) storage conditions for the 5 mg dose (4 x 1.25 mg) and 1.25 mg doses.

[0137] It is understood that the examples and embodiments described herein are for illustrative purposes and that various modifications or changes in light thereof will be suggested to those skilled in the art and are incorporated within the spirit and scope of this application and the appended claims. All publications, patents, and patent applications cited herein are incorporated by reference herein for all purposes.

Claims

1. 1. A minitablet formulation comprising bis-choline tetrathiomolybdate in an amount ranging from about 1.00 mg to about 1.50 mg (e.g., in the range of about 1.10 mg to about 1.40 mg, or about 1.15 mg to about 1.35 mg, or about 1.20 mg to about 1.30 mg, or about 1.22 mg to about 1.28 mg, or about 1.23 mg to about 1.27 mg, or about 1.24 mg to about 1.26 mg).

2. 2. The minitablet formulation of claim 1, wherein said amount of bis-choline tetrathiomolybdate is about 1.25 mg.

3. 3. The minitablet formulation of claim 1 or claim 2, further comprising about 20% to about 30% (e.g., in the range of about 22% to about 28%, or about 23% to about 27%, or about 24% to about 26%, or about 20% to about 25%, or about 25% to about 30%) of a buffer, by weight based on the weight of the minitablet core.

4. 3. A minitablet formulation according to claim 1 or claim 2, further comprising about 25% by weight of a buffer, based on the weight of the minitablet core.

5. 5. A minitablet formulation according to claim 3 or 4, wherein the buffer is sodium bicarbonate.

6. 6. The minitablet formulation of any one of claims 1 to 5, further comprising about 60% to about 70% (e.g., in the range of about 62% to about 70%, or about 63% to about 69%, or about 64% to about 68%, or about 65% to about 67%) of a filler component, by weight based on the weight of the minitablet core.

7. 7. The minitablet formulation of claim 6, further comprising about 66% by weight of a filler component, based on the weight of the minitablet core.

8. 8. A minitablet formulation according to claim 6 or 7, wherein the filler component is microcrystalline cellulose.

9. 9. The minitablet formulation according to any one of claims 1 to 8, further comprising about 0.5% to about 1% (e.g., in the range of about 0.6% to about 0.9%, or about 0.65% to about 0.85%, or about 0.7% to about 0.8%, or about 0.72% to about 0.78%, or about 0.73% to about 0.77%) of a lubricant component, by weight based on the weight of the minitablet core.

10. 10. The minitablet formulation of claim 9, further comprising about 0.75% of said lubricant component.

11. 11. A minitablet formulation according to claim 9 or 10, wherein the lubricant component is sodium stearyl fumarate.

12. 12. A minitablet formulation according to any one of claims 1 to 11, further comprising a coating on the outer surface of the formulation (e.g. the outer surface of the minitablet core comprising bis-choline tetrathiomolybdate and optionally the buffer, the filler component, and / or the lubricant component).

13. 13. The minitablet formulation of claim 12, wherein the coating comprises a seal coating, a subcoating, an enteric coating, or a combination thereof.

14. A minitablet formulation comprising: Bis-choline tetrathiomolybdate in an amount of about 1.25 mg; About 25% (by weight based on the weight of the minitablet core) of buffer, About 66% (by weight based on the weight of the minitablet core) of a filler component; A minitablet formulation comprising about 0.75% (by weight based on the weight of the minitablet core) of a lubricant component.

15. 15. The minitablet formulation of claim 14, further comprising a coating on the outer surface of said minitablet core comprising bis-choline tetrathiomolybdate, said buffer, said filler component, and said lubricant component.

16. 16. The minitablet formulation of claim 15, wherein the coating comprises a seal coating, a subcoating, an enteric coating, or a combination thereof.

17. A minitablet formulation according to any one of claims 14 to 16, wherein the buffer is sodium bicarbonate.

18. A minitablet formulation according to any one of claims 14 to 17, wherein the filler component is microcrystalline cellulose.

19. A minitablet formulation according to any one of claims 14 to 18, wherein the lubricant component is sodium stearyl fumarate.

20. 20. The minitablet formulation of any one of claims 1 to 19, wherein the minitablet formulation contains less than about 3% total impurities upon storage at about 25°C and about 60% relative humidity for 4 weeks.

21. 20. The minitablet formulation of any one of claims 1 to 19, wherein the minitablet formulation contains less than about 2% total molybdenum impurities, the molybdenum impurities being selected from one or more of TM0, TM1, TM2, and TM3 at about 25°C and about 60% relative humidity storage for 4 weeks.

22. 20. A minitablet formulation according to any one of claims 1 to 19, wherein the minitablet formulation contains about 0.7% or less of polymeric molybdenum impurity.

23. 20. The minitablet formulation of any one of claims 1 to 19, wherein the minitablet formulation contains less than about 1.3% of TM3 impurity upon storage for 4 weeks at about 25°C and about 60% relative humidity.

24. 20. The minitablet formulation of any one of claims 1 to 19, wherein the minitablet formulation contains less than about 0.3% of dimeric S6 impurity upon storage for 4 weeks at about 25°C and about 60% relative humidity.

25. A unit dose container comprising one or more minitablets according to any one of claims 1 to 24.

26. 26. The unit dose container of claim 25, containing from 2 to 24 minitablets.

27. 27. The unit dose container of claim 26, containing 2, 4, 8, 12, or 24 minitablets.

28. 28. The unit dose container of any one of claims 25 to 27, comprising a patient openable capsule, sachet or stick pack.

29. 28. A container according to any one of claims 25 to 27, comprising a unit dose dispensing container adapted to dispense unit doses of minitablets.

30. 30. The unit dose container of claim 29, wherein the unit dose dispensing container is a minitablet dispensing container.

31. 31. The unit dose container of claim 30, wherein the dispensing container is configured to dispense about 2-24 minitablets.

32. 32. The unit dose container of claim 31, wherein the dispensing container is configured to dispense 2, 4, 8, 12, or 24 unit doses of minitablets.

33. One or more minitablet formulations according to any one of claims 1 to 24 or unit dose containers according to any one of claims 25 to 32 for treating a copper metabolism related disease or disorder in a subject.

34. 34. The minitablet formulation or unit dose container of claim 33, wherein the copper metabolism related disease or disorder is Wilson's disease.

35. 35. A minitablet formulation or unit dose container according to claim 33 or claim 34, wherein the one or more minitablets or unit doses are administered daily, optionally once daily.

36. 35. A minitablet formulation or unit dose container according to claim 33 or claim 34, wherein the one or more minitablets or unit doses are administered every other day.

37. A minitablet formulation or unit dose container according to any one of claims 33 to 36, wherein the one or more minitablets or unit doses are administered in the fasted state.

38. 38. The minitablet formulation or unit dose container of any one of claims 33 to 37, wherein the amount of bis-choline tetrathiomolybdate administered is 15 mg.

39. Use of one or more minitablets according to any one of claims 1 to 24 or a unit dose according to any one of claims 25 to 32 for the manufacture of a medicament.

40. 34. Use of one or more minitablets according to any one of claims 1 to 24 or a unit dose according to any one of claims 25 to 32 for the manufacture of a medicament for treating a copper metabolism related disease or disorder in a subject.

41. 41. The use according to claim 40, wherein the copper metabolism-related disease or disorder is Wilson's disease.

42. The use according to any one of claims 39 to 41, wherein said one or more minitablets or said unit doses are administered daily, optionally once a day.

43. The use according to any one of claims 39 to 41, wherein the one or more minitablets or unit doses are administered every other day.

44. The use according to any one of claims 39 to 43, wherein the one or more minitablets or the unit dose are administered in the fasted state.

45. The use according to any one of claims 39 to 44, wherein the amount of bis-choline tetrathiomolybdate administered is 15 mg.

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