CHEN DAVID JENSON (US)
STUTZMAN TODD ANTHONY (US)
KERNER ALLISSA ROBIN (US)
SCHNITZ JOSEPH MICHAEL (US)
JAIN RAJ RAMNIK (US)
WO2019110619A1 | 2019-06-13 | |||
WO2007142626A1 | 2007-12-13 | |||
WO2019154876A1 | 2019-08-15 |
US20040019087A1 | 2004-01-29 |
What is claimed is: Claim 1. A mini-tablet formulation comprising bis-choline tetrathiomolybdate in an amount in the range of 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). Claim 2. The mini-tablet formulation of claim 1, wherein the amount of bis-choline tetrathiomolybdate is about 1.25 mg. Claim 3. The mini-tablet 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%) by weight, based on the weight of mini-tablet core, of a buffer. Claim 4. The mini-tablet formulation of claim 1 or claim 2, further comprising about 25 wt%, based on the weight of mini-tablet core, of a buffer. Claim 5. The mini-tablet formulation of claim 3 or 4, wherein the buffer is sodium bicarbonate. Claim 6. The mini-tablet formulation of any one of claims 1-5, 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%) by weight, based on the weight of mini-tablet core, of a filler component. Claim 7. The mini-tablet formulation of claim 6, further comprising about 66 wt%, based on the weight of mini-tablet core, of a filler component. Claim 8. The mini-tablet formulation of claim 6 or 7, wherein the filler component is microcrystalline cellulose. Claim 9. The mini-tablet formulation of any one of claims 1-8, 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%) by weight, based on the weight of mini-tablet core, of a lubricant component. Claim 10. The mini-tablet formulation of claim 9, further comprising about 0.75% of the lubricant component. Claim 11. The mini-tablet formulation of claim 9 or 10, wherein the lubricant component is sodium stearyl fumarate. Claim 12. The mini -tablet formulation of any one of claims 1-11 further comprising a coating on the outer surface of the formulation (e.g., an outer surface of the mini-table’s core that comprises bis-choline tetrathiomolybdate and optionally the buffer, the filler component, and/or the lubricant component). Claim 13. The mini -tablet formulation of claim 12, wherein the coating comprises a seal coating, a sub-coating, an enteric coating, or a combination thereof. Claim 14. A mini-tablet formulation comprising: bis-choline tetrathiomolybdate in an amount of about 1.25 mg; about 25% (by weight based on the weight of mini-tablet core) of a buffer; about 66% (by weight based on the weight of mini-tablet core) of a filler component; about 0.75% (by weight based on the weight of mini-tablet core) of the lubricant component. Claim 15. The mini-tablet formulation of claim 14, further comprising a coating on an outer surface of the mini-tablet’s core that comprises bis-choline tetrathiomolybdate, the buffer, the filler component, and the lubricant component. Claim 16. The mini -tablet formulation of claim 15, wherein the coating comprises a seal coating, a sub-coating, an enteric coating, or a combination thereof. Claim 17. The mini-tablet formulation of any one of claims 14-16, wherein buffer is sodium bicarbonate. Claim 18. The mini-tablet formulation of any of claims 14-17, wherein the filler component is microcrystalline cellulose. Claim 19. The mini-tablet formulation of any of claims 14-18, wherein the lubricant component is sodium stearyl fumarate. Claim 20. The mini-tablet formulation of any one of claims 1-19, wherein the mini-tablet formulation comprises no more than about 3% of total impurities at 4 weeks of storage at about 25 °C at about 60% relative humidity. Claim 21. The mini-tablet formulation of any one of claims 1-19, wherein the mini-tablet formulation comprises less than about 2%, of total molybdenum impurities, wherein the molybdenum impurities are selected from one or more of TM0, TM1, TM2, and TM3, at 4 weeks of storage at about 25 °C at about 60% relative humidity. Claim 22. The mini-tablet formulation of any one of claims 1-19, wherein the mini-tablet formulation comprises no more than about 0.7% of polymeric molybdenum impurities. Claim 23. The mini-tablet formulation of any one of claims 1-19, wherein the mini-tablet formulation comprises less than about 1.3% of TM3 impurity at 4 weeks of storage at about 25 °C at about 60% relative humidity. Claim 24. The mini-tablet formulation of any one of claims 1-19, wherein the mini-tablet formulation comprises less than about 0.3% of Dimer S6 impurity at 4 weeks of storage at about 25 °C at about 60% relative humidity. Claim 25. A unit dose container comprising one or more of the mini-tablets of any claims 1- 24. Claim 26. The unit dose container of claim 25 comprising from 2 to 24 mini-tablets. Claim 27. The unit dose container of claim 26, comprising 2, 4, 8, 12, or 24 mini-tablets. Claim 28. The unit dose container of any one of claims 25-27 comprising a capsule that can be opened by the patient, a sachet, or a stick pack. Claim 29. The container of any one of claims 25-27 comprising a unit dose dispenser configured to dispense a unit dose of mini-tablets. Claim 30. The unit dose container of claim 29, wherein the unit dose dispenser is a mini- tablet dispenser. Claim 31. The unit dose container of claim 30, wherein the dispenser is configured to dispense about 2 to 24 mini-tablets. Claim 32. The unit dose container of claim 31, wherein the dispenser is configured to dispense a unit dose of 2, 4, 8, 12, or 24 mini-tablets. Claim 33. A method for treating a copper metabolism-associated disease or disorder in a subject, the method comprising administering to the subject one or more mini-tablets of any claims 1-24 or a unit dose as described in any of claims 25-32. Claim 34. The method of claim 33, wherein the copper metabolism-associated disease or disorder is Wilson Disease. Claim 35. The method of claim 33 or claim 34, wherein the one or more mini-tablets or the unit dose is administered daily, optionally once daily. Claim 36. The method of claim 33 or claim 34, wherein the one or more mini-tablets or the unit dose is administered every other day. Claim 37. The method of any claims 33-36, wherein the one or more mini-tablets or the unit dose is administered in fasted state. Claim 38. The method of any claims 33-37, wherein the amount of bis-choline tetrathiomolybdate administered is 15 mg. Claim 39. Use of one or more mini-tablets of any claims 1-24 or a unit dose as described in any of claims 25-32 for the manufacture of a medicament. Claim 40. Use of one or more mini-tablets of any claims 1-24 or a unit dose as described in any of claims 25-32 for the manufacture of a medicament for treating a copper metabolism- associated disease or disorder in a subject. Claim 41. The use of claim 40, wherein the copper metabolism-associated disease or disorder is Wilson Disease. Claim 42. The use of any of claims 39-41, wherein the one or more mini-tablets or the unit dose is administered daily, optionally once daily. Claim 43. The use of any of claims 39-41, wherein the one or more mini-tablets or the unit dose is administered every other day. Claim 44. The use of any of claims 39-43, wherein the one or more mini-tablets or the unit dose is administered in fasted state. Claim 45. The use of any of claims 39-44, wherein the amount of bis-choline tetrathiomolybdate administered is 15 mg. |
Example 2: Accelerated 4 Week Stability of Mini-Tablets [0052] The objective of the stability study was to assess the stability profile of several of BC- TTM mini-tablet formulations. The stability was evaluated using observation of one tablet (for product appearance) and HPLC/UV (200 to 400 nm) analysis of injection from one tablet sample preparation (for assay of BC-TTM and impurities content). The stability of the mini- tablets was compared to a tablet comprising 5 mg of ALX1840, having a formulation as shown in Table 3. [0053] The stability of the mini-tablets of the disclosure was evaluated at start (“ATST”), at week 1 (“1W”), at week 2 (“2W”), and at week 4 (“4W”) when stored at 5°C, at 25°C at 60% relative humidity (RH), and 40°C at 75%RH. Table 4 provides evaluation of Formulation #1, Generation 2 (F1G2) of Example 1; Table 5 provides evaluation of Formulation #2, Generation #2 (F2G2) of Example 1; and Table 6 provides evaluation of 5mg tablet. LTLOQ as used herein means “lower than limit-of-quantification”; ND as used herein means “not determined.” For Tables 4-7 and 10, the reported amounts of TM0 were measured as TM0 in its anion form ([MoO 4 ] ^^ ), whereas the TM0 in the remainder of the disclosure is reported in terms of its choline salt form. The “Total Impurities” amounts reported in Tables 4-7 and 10, therefore, were calculated using the amount of TM0 in its anion form, whereas the “Total Impurities” amounts reported in the remainder of the disclosure were calculated using the amount of TM0 in its choline salt form. [0054] Surprisingly, the 1.25 mg F2G2 mini-tablet showed greater stability compared to the 5 mg tablet as illustrated by the lower concentration of total impurities (%) over time; and the higher concentration of BC-TTM (%) over time (Figures 1 and Table 7). In addition, Figure 2 illustrates that the 1.25 mg F2G2 mini-tablet also showed greater stability compared to the 1.25 mg F1G2 mini-tablet. Table 3: Formulation of 5 mg BC-TTM Tablet
Example 3: Manufacturing Mini-Tablets [0055] Mini-tablets of the disclosure were prepared on a manufacturing scale. The batch formula for the mini-tablet is provided in Table 8 below. Smaller or larger batches using the components and proportions may be produced. The mini-tablets cores were prepared using a dry-granulation process. In short, upon final blending, mini-tablet cores were produced using a compression machine to match its targeted physical attributes. Subsequently, mini- tablet cores were subject to seal coating, sub-coating, and finally enteric coating. The mini- tablet manufacturing processes used commercially available pharmaceutical processing equipment commonly used for the manufacturing of tablet dosage forms. Table 8: Drug Product Batch Formula 1 Drug substance quantity may be adjusted based on lot specific potency and the difference adjusted with Microcrystalline Cellulose, NF quantity. 2 The actual quantity will be adjusted based on the actual yield of the milled granules. 3 Coating operations performed in three sub-lots of approximate equal size. 4 Water amount used for preparation of coating dispersions of Opadry 200 Clear and Acryl- EZE White may subject to adjustment based on the batch size and is not part of the finished product except for the residual amount remaining after drying. Description of Manufacturing Process and Process Controls [0056] The manufacturing process consisted of compounding of drug substance and excipients in a dry granulation process. The final blend was then compressed into mini-tablet cores. Coating processes started with a seal coating of the cores with Carnauba Wax. Then the seal coated tablets were subject to sub-coating using Opadry 200 Clear followed by enteric coating with Acryl-EZE White. The major processing steps are pre-roller compaction blending, roller compaction and milling of the ribbons, final blending of bulk granules with extragranular excipient, mini-tablet compression, seal coating, sub coating and enteric coating. [0057] Pre-Roller Compaction Blending: Prior to processing, it was confirmed for each batch that the BC-TTM had been dispensed within two days of the manufacturing start date. Sodium Bicarbonate, USP Grade 1 Powder – Increment 1 was charged into a 15L bin then BC-TTM was added into the same 15L bin. The bag containing the residual BC-TTM was rinsed with Sodium Bicarbonate, USP Grade 1 Powder – Increment 2 and then added into the same 15L bin. Then these materials were blended in the 15L bin for 5 minutes at 10 RPM. Then Sodium Bicarbonate, USP Grade 1 Powder – Increment 3 was charged into the 15L bin and the materials were blended for 10 minutes at 10 RPM. [0058] The blended components were then discharged into interim containers and then de- lumped using a Quadro Comil equipped with a 032R screen (~812 microns). The de-lumped materials then were charged back into the same 15L bin and mixed for an additional 5 minutes at 10 RPM. Microcrystalline Cellulose, NF (Avicel PH-112) was de-lumped by passing it through the same Comil fitted with 032R screen and collected in a clean suitable container. The de-lumped Microcrystalline Cellulose, NF (Avicel PH-112) was charged into the same 15L bin and mixed for 15 minutes at 10 RPM. [0059] An equal volume of blend from the 15L bin was added to the Sodium Stearyl Fumarate, NF (Intragranular) and mixed by inverting the bag for approximately 20 seconds. This mixture was co-screened through a 20 mesh hand screen directly into the 15L bin and blended for 5 minutes at 10 RPM. The pre-roller compaction blend was then discharged into an interim container and the yield and accountability was calculated. [0060] Roller Compaction and Milling of Ribbons: The pre-compaction blend was roller compacted using the Alexanderwerks WP120 roller compactor equipped with 40 mm upper smooth/lower square rollers and a chiller set at 15°C. The ribbons were milled using the integrated inline mill on the Alexanderwerks WP120 roller compactor fitted with 1.0 mm coarse screen and 0.63 mm fine screen at 95 RPM. [0061] Ribbon and milled granule samples were collected from the beginning, middle and end of roller compaction. Upon completion of roller compaction, the milled granules were collected into an interim container for immediate continuation of processing. [0062] Final Blending: Based on the yield of granules collected from the roller compaction and milling step, the weights of the extragranular component (Sodium Stearyl Fumarate, NF) was adjusted. Initially about 50% of the milled granules were charged into a 15L bin. An equal volume of the milled granules from the remaining granules was added to the Sodium Stearyl Fumarate, NF (Extragranular) and mixed by inverting the bag for approximately 20 seconds and then hand screened through a 20 mesh screen directly into the bin, and then charged the remaining milled granules were charged into the bin. The mix was blended at 10 RPM for 5 minutes. [0063] Final blend uniformity samples were collected from ten (10) locations in triplicate from the bin using a disposable 0.5ml sample thief. An approximately 100 g sample from the bin was also collected and then the final blend was discharged into a foil bag, double lined with polyethylene bags with one desiccant in the headspace of the outer polyethylene bag. Air was removed from the polyethylene bags prior to closure with zip ties. Similarly, air was removed from the foil bag and then purged with nitrogen for approximately 3 minutes prior to heat sealing. The yield and accountability were calculated. The foil bag was then placed into a foil-lined fiber drum and returned to 2-8°C storage. [0064] Mini-tablet Core: The BC-TTM Final Blend (8.33 % by weight based on the weight of the core) was compressed into mini-tablets cores using a Korsch XL 100 Pro Tablet Press equipped with 3mm Round Multi Tip tooling and the force feeder. The compressed tablets were dedusted using a Key tablet deduster and metal checked using a Lock Met30+ Metal Detector. The mini-tablets were compressed to a target weight of 15 mg/unit and complying with other physical attributes. In-process samples were collected and tested for physical attributes at predetermined time interval during compression to ensure product quality. [0065] Bulk core tablets were collected into a foil bag, double lined with polyethylene bags with one desiccant in the headspace of the outer polyethylene bag. As much air as possible was removed from the polyethylene bags prior to closure with zip ties. The foil bag also went through the process to remove air as much as possible and then it was purged with nitrogen for approximately 3 minutes prior to heat sealing. The foil bag was then placed into a foil- lined fiber drum and returned to 2-8°C storage. [0066] Enteric Coating of Mini-tablets: Three sub-lots, with almost equal pan load size and identical coating process, are required to coat the whole theoretical batch. [0067] A seal-coat coating of Carnauba Wax, NF Powdered #1 was applied on to the mini- tablets cores using a pan-coating system. Core mini-tablets were seal coated in a Compu- Lab coater fitted with a 15” pan to a theoretical weight gain of 1%. [0068] A sub-coat coating dispersion was prepared at 20% solid content using Opadry 200 Clear (203A190001) coating system and purified water. Core tablets were sub coated in a Compu-Lab coater fitted with 15” pan to a theoretical weight gain of 20% ± 1%. [0069] An enteric coat coating dispersion was prepared at 20% solid content using Acryl- EZE White coating system and purified water. Sub coated tablets were coated in a Compu- Lab coater fitted with 15” pan to a theoretical weight gain of 35% ± 1%. [0070] Upon coating completion, the bulk enteric-coated tablets were collected in a foil bag, double lined with polyethylene bags with one desiccant in the headspace of the outer polyethylene bag. As much air as possible was removed from the polyethylene bags prior to closure with zip ties. The foil bag also had as much air as possible removed and then it was purged with nitrogen for approximately 3 minutes prior to heat sealing. The foil bag was then placed into a foil-lined fiber drum and returned. [0071] A summary of the drug product manufacturing in-process controls is provided in Table 9. [0072] There were no significant differences between 1.25 mg mini-tablet formulations produced by different batches (Table 10). Table 10: Comparison Between Batch 1 and Batch 2 Mini-Tablet Samples Example 4: Six-Month Stability of Capsules Comprising Low Dose Formulation [0073] Mini-tablets prepared according to Example 3 were placed in hydroxypropyl methylcellulose (HPMC) sprinkle capsules. Each HPMC capsule contained four (4) individual 1.25 mg mini-tablets. The capsules were stored in 60 cc HDPE WM round bottle ((0060Hl- 01) (33/400) Q024847) closed with DPC CRH1110033MM WHT SECURX RIBD SIDE PP CRC TXT (7821H1-G1263131). Each bottle contained 30 capsules. [0074] The stability of the capsules was evaluated based on product appearance, assay/impurities, dissolution, and moisture when stored at 5°C and at 25°C/60%RH. The stability data measured at 0, 1, 2, 3, 4, 5, 6, and 12 months is provided in Tables 11 and 12 for samples stored at 5°C and 25°C/60%RH conditions, respectively. LTLOQ as used herein means “lower than limit-of-quantification”; ND as used herein means “not determined.”
[0075] Another set of capsules (4 mini-tablets per capsule, prepared and stored as noted above, except that the bottles were closed with 33mm SCRX RIBD SIDE WHT PP CRC TXT TOP (HS130-357903Hl-1Cl 263455)) containing another batch of 1.25 mg mini-tablets prepared according to Example 3 (a so-called “second batch”) were also tested for long-term stability, relative to the standards provided in Table 14. Table 13 provides the results of the stability evaluation at 3 months of storage at 5°C and at 25°C/60%RH; 6 months of storage at 25°C/60%RH; and12 months of storage at 5°C. Table 13: 3, 6, and 12-Month Stability Data for Second Batch of 1.25 mg Mini-Tablets Table 14: Stability Testing Standards Example 5: Relative Bioavailability of Two Oral Formulations of BC-TTM in Healthy Adult Participants [0076] A phase 1, randomized, 2-period, 2-sequence, crossover with parallel-group extension, open -label study was conducted to compare the relative bioavailability of 2 oral formulations of BC-TTM in healthy adult participants. The purpose of this study was to assess relative bioavailability of the 1.25 mg enteric-coated (EC) mini-tablet formulation of BC-TTM compared with a 15 mg EC tablet of BC-TTM to assess 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 mini-tablet doses. The 15 mg EC tablet of BC-TTM used in the study had a formulation consisting of the components listed in Table 3. The 1.25 mg EC mini-tablets of BC-TTM were prepared in accordance with Example 3 and the drug product batch formula of Table 8. [0077] This was a 2-period, 2-sequence crossover study with parallel group extension designed to assess the relative bioavailability of equal doses of BC-TTM administered as 1.25 mg EC mini-tablets versus a single 15 mg EC tablet, and to assess 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 mini-tablet doses in the Dose-Proportionality Extension Period. The safety and tolerability of the 2 formulations of BC-TTM in healthy participants was also assessed. BC-TTM pharmacokinetics (PK) in plasma as measured via total molybdenum (Mo) and plasma ultrafiltrate (PUF) Mo was determined.
Table 15: Objectives and Endpoints of Study
[0078] The study had a Screening Period (Days -28 to -2), the Two-way Crossover Period, consisting of 2 dosing periods (Day 1 to Day 11 each), and a Dose-Proportionality 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 in Dosing Period 1. If discharged after Dosing Period 1, participants were readmitted to the CRU for Dosing Period 2 following a minimum washout of 14 days after the previous dose, and again for the Dose- Proportionality Extension Period after a minimum washout of 14 days. The end of study (EOS) visit took place 14 days (± 2 days) after the dose of BC-TTM in the Dose- Proportionality Extension Period. [0079] The Two-way Crossover Period was a randomized, open-label, 2-way (2-period, 2- sequence), crossover design to assess the relative bioavailability of 12 × 1.25 mg EC mini- tablets compared with the 15 mg EC tablet currently used in clinical studies. Participants were randomized to one of the two treatments sequences. Randomized treatment assignment were based on Baseline body mass index (BMI). Two strata for BMI (< 25, 25 to < 32 kg/m 2 ) were used: x Treatment A: BC-TTM 12 × 1.25 mg EC mini-tablets x Treatment B: BC-TTM single 15 mg EC tablet (reference tablet, currently being tested in the Phase 3 Study WTX101-301) Blood samples for PK analysis of total and PUF Mo (as surrogate measures of BC-TTM PK) and pharmacodynamic (PD)/biomarkers were collected in each dosing period on Day 1 at pre-dose, and postdose at 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours (Day 2) and then at 24 hour intervals on Days 3, 4, 5, 6, 7, 8, 9, 10, and 11. [0080] The 336-hour sample for Dosing Period 1 were collected predose in Dosing Period 2. Participants could have been discharged on Day 11 of each dosing period after completion of all procedures and review of all safety data. The end of Dosing Period 2 occurred on Day 15 ± 2 of Dosing Period 2, with the collection of the 336-hour PK sample for Dosing Period 2. [0081] The Dose-Proportionality Extension Period was a re-randomized, open-label, parallel group design to assess the dose-proportionality between 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 mini-tablet doses. The 15 mg (12 × 1.25 mg) dose was not repeated during the Dose-Proportionality Extension Period. [0082] The Dose-Proportionality Extension Period was conducted following completion of the Two-way Crossover Period of the study and after an at least 14-day washout period. Participants were re-randomized as follows: x Treatment C (N=10-12): BC-TTM 2.5 mg (2 × 1.25 mg EC mini-tablets) x Treatment D (N=10-12): BC-TTM 5 mg (4 × 1.25 mg EC mini-tablets) x Treatment E (N=10-12): BC-TTM 10 mg (8 × 1.25 mg EC mini-tablets) x Treatment F (N=10-12): BC-TTM 30 mg (24 × 1.25 mg EC mini-tablets) [0083] The dose-proportionality evaluation included data obtained from Treatment A of the Two-way Crossover Period (12 × 1.25 mg EC mini-tablets) to represent a dose of 15 mg. [0084] Re-randomized treatment assignment were based on Baseline body mass index (BMI). Two strata for Baseline BMI (< 25, 25 to < 32 kg/m 2 ) were used. Block randomization was used to equally randomly assign participants to each treatment. [0085] Participants could have been discharged on Day 11 of the Dose-Proportionality Extension Period after completion of all procedures and review of all safety data. [0086] Participants could have been asked or required to stay in the CRU during the Two- way Crossover Period, and/or at the end of the Dose-Proportionality Extension Period before the end of study (EOS) visit, for their own safety, and also to maintain the integrity of the conduct of the study. [0087] The final data showed that of the 48 randomized participants, 44 participants completed the Two-way Crossover Period and 40 participants completed the Dose- Proportionality Extension Period. All 48 (100%) participants randomized in the Two-way Crossover Period were included in the Safety, PKDS-CO, and Full Analysis sets, and all 41 (100%) participants randomized in the Dose-Proportionality Extension Period were included in the Safety, PKDS-E, and Full Analysis sets. [0088] Plasma total and PUF molybdenum as surrogate measures for BC-TTM PK profiles and PK parameters were comparable between a single dose of BC-TTM administered as 12 × 1.25 mg EC mini tablets (15 mg total dose) and as 1 × 15 mg EC tablet. There were no clinically relevant differences between the 2 treatment formulations under fasting conditions in healthy participants as the 90% CIs for C max , AUC t and AUC ^ of total molybdenum were contained within the 80% to 125% bioequivalence limits.
Table 16: Summary of PK Parameters of Plasma Total and PUF Molybdenum - Two-way Crossover Period a PK parameters were calculated based on corrected concentrations and all parameter YDOXHV^^H[FHSW^IRU^^ z ) are rounded to one digit after decimal point from source data b Data presented as mean ± SD (%CV) except for t max and t lag as median (range). c Molybdenum dose was used to calculate CL/F or V d /F values. [0089] Plasma total molybdenum PK parameters generally showed a dose-proportional increase from 2.5 mg to 30 mg for the BC-TTM EC mini-tablet formulation. Plasma PUF molybdenum PK parameters showed a less than dose-proportional increase from 2.5 mg to 30 mg for the BC-TTM EC mini-tablet formulation. BC-TTM PK were apparently not affected by body weight or BMI. Table 17: Summary of PK Parameters of Plasma Total and PUF Molybdenum - Dose- Proportionality Extension Period a PK parameters were calculated based on corrected concentrations and all parameter YDOXHV^^H[FHSW^IRU^^ z ) are rounded to one digit after decimal point. b Data presented as mean ± SD (%CV) except for t max and t lag as median (range). c Molybdenum dose was used to calculate CL/F or V d /F values. [0090] Dose-normalized plasma total molybdenum C max and AUC t values decreased moderately with increasing dose, with a decrease more prominent in AUC ^ values. For PUF molybdenum, dose-normalized plasma exposure values decreased with increasing dose, indicating that BC-TTM PUF molybdenum exposure increased in a less than dose proportional manner within the BC-TTM dose range of 2.5 mg to 30 mg for the EC mini- tablet formulation (Table 18). Table 18: Summary of Dose-normalized PK Parameters of Plasma Total and PUF Molybdenum - Dose-Proportionality Extension Period (PKDS-E Set and Treatment A from PKDS-CO Set) a PK parameters were calculated based on corrected concentrations and all parameter values are rounded to one digit after decimal point from the source data. [0091] The results of the analyses for a potential formulation difference between Treatments A and B indicate that there were no clinically meaningful differences in BC-TTM PK parameters between the 2 treatments or formulations. Plasma total molybdenum (C max , AUC t , and AUC ^ ) and PUF molybdenum (C max ) geometric mean ratios (90% CI) were contained entirely within the default no-effect 90% CI boundary of 80% to 125%, except for PUF molybdenum AUC t where geometric mean ratio (90% CI) was 101.2% (70.6% to 145.1%), with the lower and upper boundary marginally extending outside of the no-effect boundary of 80% to 125% (Table 19). Table 19: Relative Bioavailability of Plasma Total and PUF Molybdenum (PKDS- CO Set) PK parameters were calculated using corrected concentrations. b Bioavailability was derived using an ANOVA statistical model with dosing period, treatment, and treatment sequence as the fixed effects and the participant as a random effect, using the natural logarithms of the data. Bioavailability was then defined as the ratio of the geometric means of PK parameter (Cmax, AUCt, and AUC-) for the test (12 x 1.25 mg BC- TTM EC mini-tablets) over the reference (1 x 15 mg BC-TTM EC tablet) treatment.
[0092] Total molybdenum: For the 2.5 mg to 15 mg dose range and the 2.5 mg to 30 mg dose range, dose-proportionality criteria for Cmax and AUCt were met as 90% Cl slope values fell inside the critical intervals defined as ([1+ln(0.5)/ln(p), 1+ln(2)/ln(p)]). However, for the 2.5 mg to 5 mg dose range and the 2.5 mg to 10 mg dose range, the dose-proportionality criteria for Cmax and AUCt were not met. For AUC-, the dose-proportionality criterion was met only for the 2.5 mg to 10 mg dose range, but was not for the 2.5 mg to 15 mg dose range and the 2.5 mg to 30 mg dose range. Overall, the power model based dose-proportionality analysis results demonstrate that increases in total molybdenum exposure are generally dose proportional across the investigated dose range of 2.5 mg to 30 mg. [0093] PUF molybdenum: The dose-proportionality criteria for C max and AUC t values were not met for any dose range. The power model-based dose-proportionality analysis results demonstrate that increases in PUF molybdenum exposure were less than dose proportional across the investigated dose range of 2.5 mg to 30 mg due, most likely, to the much higher variability in the PUF molybdenum concentrations versus plasma total molybdenum. Table 20: Power Model Assessment of Dose-Proportionality of Plasma Total and PUF Molybdenum (PKDS-E Set and Treatment A from PKDS-CO Set) * Dose proportionality criteria was met as the 90% Cl values were contained entirely within the critical interval defined as ([1 + ln(0.5)/ln(p), 1 + ln(2)/ln(p)]), dose-proportionality was supported across the investigated dose range. a PK parameters were calculated based on corrected concentrations. b Equivalent molybdenum dose was used in the power model dose-proportionality analysis.
[0094] There were no apparent differences in BC-TTM RD parameters (plasma total and RUF copper concentrations) between 12 * 1.25 mg EC mini-tablets and the 15 mg reference EC tablet Maximum plasma total copper concentration occurred 8 hours post-dose and then gradually decreased and eventually returned to pre-dose Baseline concentrations by 96 to 120 hours post-dose. The pre-dose Baseline mean plasma total copper concentration of Treatments A and B were 988 and 986 ng/mL, respectively, and transiently increased to a mean maximum of 1230 and 1210 ng/mL, respectively, at 8 hours post-dose.
[0095] After the 8-hour post-dose time point, plasma total copper concentrations gradually decreased, and the mean concentration declined to < 11% above the pre-dose Baseline at 48 hours post-dose. By 96 to 120 hours post-dose, total copper concentrations had returned to pre-dose Baseline concentrations. RUF copper concentrations were much lower than total copper concentrations (mean value of less than 10 ng/mL) at all sampling time points limiting the opportunity for quantitative assessments.
[0096] Summary statistics for absolute and percentage change from Baseline plasma total copper concentrations following Treatments C, D, E, and F were calculated. Treatment A from the Two-way Crossover Period was included for comparison. For Treatment C (2.5 mg BC-TTM, lowest BC-TTM dose), plasma total copper concentration versus time profiles remained stable overall. The plasma total copper concentration versus time profiles following Treatments D, E, and F showed a similar trend as the profiles of Treatments A and B. Plasma total copper concentrations reached a maximum at 6 to 12 hours post-dose and centered around 8 hours, with a maximum mean percentage change (increase) from Baseline (0.5 hours pre-dose) of approximately 2%, 10%, 18%, 26%, and 31% for Treatments C, D, E, A, and F, respectively. The increases of maximum mean percentage changes are dose dependent, but less than dose proportional.
[0097] After the 12-hour post-dose time point, plasma total copper concentrations gradually decreased with the median percent change from Baseline reaching within approximately < 15% of the pre-dose Baseline at 48 hours post-dose. At 120 to 144 hours post-dose, total copper concentrations had returned to pre-dose Baseline levels. RUF copper concentrations were much lower than total copper concentrations (mean value of less than 10 ng/mL) at all sampling time points, limiting the opportunity for quantitative assessments [0098] BC-TTM had an acceptable safety profile and was generally well-tolerated in healthy adult participants when administered as a single oral dose from 2.5 mg to 30 mg as EC mini-tablets and as a 15 mg EC tablet 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 2 events of increased blood creatine phosphokinase blood concentrations of Grade 4 severity reported by 2 (4.3%) participants following Treatment B during the Two- way Crossover Period. The incidence of TEAEs was similar between Treatment A (BC-TTM 12 × 1.25 mg EC mini-tablets) and Treatment B (BC-TTM single 15 mg EC reference tablet), and no dose-relationship was observed for the Treatments C to F (2.5 mg to 30 mg BC-TTM administered as 1.25 mg EC mini-tablets). Example 6. Food Vehicle Study [0099] The food study was performed to observe and test the integrity and stability of the BC-TTM 1.25-mg mini-tablets once introduced to a food vehicle. The BC-TTM 1.25-mg mini- tablets were prepared in accordance with Example 3 and the drug product batch formula of Table 8. The mini-tablets were tested at a 5-mg (4 x 1.25-mg) dose and a 1.25-mg dose in either yogurt or applesauce. The samples were allowed to soak in the food vehicles for allotted time-points at both room temperature and 5°C food storage conditions. The samples were then removed from the food vehicles for visual observations and tested. [0100] Specifically, the study was conducted as follows: 1. Samples were tested at n=3. 2. Both room temperature and 5°C storage conditions of the food vehicles were tested to determine if the storage of the food vehicle had an influence on the integrity of the sample. 3. Samples were tested at a 5-mg dose (4 x 1.25-mg) and a 1.25-mg dose. 4. Applesauce Soaking Time-Points: 5, 7.5, 10, 12.5, and 15 minutes. 5. Yogurt Soaking Time-Points: 5, 10, 15, 30, 45, 60, 90, and 120 minutes. 6. Delivery technique: mini-tablets were placed on top of the food vehicle and stirred in from top to bottom a total of three times to best represent the handling likely during administration. 7. Food vehicles were not added to dissolution vessels following sample soaking. [0101] Yogurt [0102] Yoplait Original French Vanilla Low Fat Yogurt (6oz) (pH 4.24) was the brand used for the yogurt food vehicle. [0103] A 5-mg dose (4 x 1.25-mg) or a 1.25-mg dose was placed on top of the yogurt and a spoon was used to stir in the mini-tablet(s) from bottom to top a total of three times, ensuring the samples were fully covered. The spoon was then removed and the foil-lid was placed over to cover. The mini-tablets were allowed to soak in the yogurt for the following time-points: 5, 10, 15, 30, 45, 60, 90, and 120 minutes. The samples for each time-point were tested at n=3, at both 5°C and room temperature food vehicle storage conditions. The room temperature samples were left on the lab countertops for the duration of the food soaking, whereas the 5°C samples were immediately placed into 5°C storage after introduction to the yogurt. After the allotted time-points, the samples were removed from the yogurt and observed. [0104] For the 5 mg dose, the mini-tablets were placed into dissolution apparatus 1 baskets and transferred to an acid stage bath (500 mL, 0.1 N HCl, 37°C ± 0.5°C) for two hours set to a rotation speed of 100 rpm. The samples were then removed from the acid bath for observation and transferred to a buffer stage bath (500 mL, modified Simulated Intestinal Fluid pH 7.5 ± 0.05, 37°C ± 0.5°C) set to a rotation speed of 75 rpm. Samples were taken at 10, 12.5, 15, 20, and 30 minutes. Following the 20 minutes sampling time- point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC. [0105] For the 1.25 mg dose, the mini-tablet was placed into a dissolution apparatus 2 mini-vessel acid stage bath (75 mL, 0.1 N HCl 37°C ± 0.5°C) for two hours set to a rotation speed of 100 rpm. Following the two-hour acid stage, the mini-tablet was observed and a buffer solution was added to the vessel (25 mL, 0.25M Tribasic Sodium Phosphate, pre- heated to 37°C ± 0.5°C). The paddle speed rotation was decreased to 75 rpm, and samples were taken at 10, 12.5, 15, 20, and 30 minutes. Following the 20 minutes sampling time- point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC. [0106] For the 5-mg dose (4 x 1.25-mg), the samples tested in yogurt showed no visible signs of swelling or discoloration throughout testing. The integrity of the mini-tablets was not compromised by the introduction to yogurt. [0107] For the 1.25 mg dose, the samples tested in yogurt showed no visible signs of swelling or discoloration throughout testing. The integrity of the mini-tablets coating was not compromised by the introduction to yogurt. [0108] Applesauce [0109] Mott’s Applesauce (4oz) (pH 3.68) was the brand used for the applesauce food vehicle. [0110] A 5-mg dose (4 x 1.25-mg) or 1.25-mg dose was placed on top of the applesauce and a spoon was used to stir in the mini-tablets from bottom to top a total of three times, ensuring the samples were fully covered. The spoon was then removed and the foil-lid was placed over to cover. The mini-tablets were allowed to soak in the applesauce for the following time-points: 5, 7.5, 10.12.5, and 15 minutes. The samples for each time-point were tested at n=3, at both 5°C and room temperature food vehicle storage conditions. The room temperature samples were left on the lab countertops for the duration of the food soaking, whereas the 5°C samples were immediately placed into 5°C storage after introduction to the applesauce. After the allotted time-points, the samples were removed from the food vehicle and observed. [0111] For the 5 mg dose, the mini-tablets were placed in dissolution apparatus 1 baskets and transferred to an acid stage bath (500 mL, 0.1 N HCl, 37°C ± 0.5°C) for two hours set to a rotation speed of 100 rpm. The samples were then removed from the acid bath for observation and transferred to a buffer stage bath (500 mL, modified Simulated Intestinal Fluid pH 7.5 ± 0.05, 37°C ± 0.5°C) set to a rotation speed of 75 rpm. Samples were taken at 10, 12.5, 15, 20, and 30 minutes. Following the 20 minutes sampling time- point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC. [0112] For the 1.25 mg dose, the mini-tablet was placed into a dissolution apparatus 2 mini-vessel acid stage bath (75 mL, 0.1 N HCl 37°C ± 0.5°C) for two hours set to a rotation speed of 100 rpm. Following the two-hour acid stage, the mini-tablet was observed and a buffer solution was added to the vessel (25 mL, 0.25M Tribasic Sodium Phosphate pre- heated to 37°C ± 0.5°C). The paddle speed rotation was decreased to 75 rpm, and samples were taken at 10, 12.5, 15, 20, and 30 minutes. Following the 20-minute sampling time-point, the rotation speed was increased to 250 rpm. The samples were then analyzed using HPLC. [0113] For the 5 mg does, following the soaking in the applesauce, the mini-tablets were observed and there were no visible signs of discoloration or degradation. All samples were then moved to the two-hour acid stage bath. Following the two-hour acid stage, the mini- tablets were observed. All 5°C time-point samples remained intact, with no signs of swelling or discoloration. The room temperature 5 and 7.5 minute time-point samples also remained intact, with no visible signs of discoloration or degradation. All the room temperature time- point samples following 7.5 minutes (10, 12.5, and 15 minutes) had degraded in the acid stage. Only the 5°C samples and the 5 and 7.5 minute time-point room temperature samples were able to continue to the buffer stage. [0114] For the 1.25 mg does, following the soaking in the applesauce, the mini-tablet was observed and there was no visible sign of discoloration or degradation. All samples were then moved to the two-hour acid stage bath. Following the two-hour acid stage, the mini-tablets were observed. All 5°C time-point samples remained intact, with no signs of swelling or discoloration. The room temperature 5 and 7.5 minute time-point samples also remained intact, with no visible signs of swelling or degradation. All the room temperature time-point samples following 7.5 minutes (10, 12.5, and 15 minutes) showed signs of slight swelling throughout the acid stage, but no visible signs of discoloration or degradation. [0115] The results summarized in this example confirm that BC-TTM 1.25-mg enteric coated mini-tablets have stability after introduction to a food vehicle. In applesauce, the mini- tablets are stable for up to 7.5 minutes at room temperature, and up to 15 minutes at 5°C (refrigerated) for 5-mg doses (4 x 1.25-mg) and up to 15 minutes at both room temperature and 5°C (refrigerated) storage conditions for 1.25-mg doses. In yogurt, the mini-tablets are stable for up to 120 minutes at both room temperature and 5°C (refrigerated) storage conditions for 5-mg (4 x 1.25-mg) and 1.25-mg doses. [0116] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof are suggested to persons skilled in the art and are to be incorporated within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated herein by reference for all purposes.