Methods for screening for VMAT2 inhibitors

Abstract

The present application relates to a method for preparing a pharmaceutical composition comprising a VMAT2 inhibitor, and / or a method for identifying a therapeutically effective dose of a VMAT2 inhibitor, which is useful for treating neurological and psychiatric diseases and disorders, and achieving 80-96% occupancy in subjects. Dysregulation of the dopaminergic system is essential for several central nervous system (CNS) disorders, including neurological and psychiatric diseases and disorders.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 235,407, filed August 20, 2021, the disclosure of which is incorporated by reference in its entirety herein.

[0002] Technical Field The present application relates to a method for preparing a pharmaceutical composition comprising a VMAT2 inhibitor, and / or a method for identifying a therapeutically effective dose of a VMAT2 inhibitor, which is useful for treating neurological and psychiatric diseases and disorders, and achieving an occupancy rate of 80-96% in subjects. [Background technology]

[0003] background Dysregulation of the dopaminergic system is essential for several central nervous system (CNS) disorders, including neurological and psychiatric diseases and disorders. These neurological and psychiatric diseases and disorders include hyperkinetic movement disorders and conditions, such as schizophrenia and mood disorders. The transporter protein vesicular monoamine transporter-2 (VMAT2) plays a key role in presynaptic dopamine release, regulating monoamine uptake from the cytoplasm into synaptic vesicles for storage and release. Summary of the Invention [Means for solving the problem]

[0004] overview The present application relates, inter alia, to a method for preparing a pharmaceutical composition comprising a therapeutically effective dose of a VMAT2 inhibitor, the method comprising: (a) administering to a subject an amount of a VMAT2 inhibitor; (b) measuring in vivo VMAT2 occupancy of the VMAT2 inhibitor in the subject, wherein a VMAT2 occupancy of 80-96% indicates that the amount of the VMAT2 inhibitor is a therapeutically effective dose; and (c) combining a therapeutically effective dose of a VMAT2 inhibitor with a pharma- ceutically acceptable carrier. The present invention provides a method comprising:

[0005] The present application provides a method for identifying a therapeutically effective dose of a VMAT2 inhibitor, comprising: (a) administering to a subject an amount of a VMAT2 inhibitor; (b) measuring the in vivo VMAT2 occupancy of a VMAT2 inhibitor in a subject; and (c) identifying a therapeutically effective dose of the VMAT2 inhibitor, where the VMAT2 occupancy of that amount of the VMAT2 inhibitor is between 80% and 96%. The present invention further provides a method comprising:

[0006] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Methods and materials for use in the present invention are described herein; other suitable methods and materials known in the art can also be used.Materials, methods, and examples are illustrative only and are not intended to be limiting.All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety.In case of discrepancy, the present specification, including definitions, will prevail. [Brief description of the drawings]

[0007] [Figure 1A]Figures 1A-1C show [18F]AV-133 SUV images averaged at baseline and 60-120 min after various doses of NBI-750142 in three different cynomolgus monkeys: GC786 (Figure 1A), EC865 (Figure 1B), and LC206 (Figure 1C). A dose-dependent decrease in striatal uptake was observed. [Figure 1B] Figures 1A-1C show [18F]AV-133 SUV images averaged at baseline and 60-120 min after various doses of NBI-750142 in three different cynomolgus monkeys: GC786 (Figure 1A), EC865 (Figure 1B), and LC206 (Figure 1C). A dose-dependent decrease in striatal uptake was observed. [Figure 1C] Figures 1A-1C show [18F]AV-133 SUV images averaged at baseline and 60-120 min after various doses of NBI-750142 in three different cynomolgus monkeys: GC786 (Figure 1A), EC865 (Figure 1B), and LC206 (Figure 1C). A dose-dependent decrease in striatal uptake was observed. [Figure 1D] FIG. 1D shows a cynomolgus monkey MR image for anatomical reference when comparing the data in FIGS. 1A-1C. [Figure 2-1] FIG. 2 shows [ 18 F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-750142 from the NHP model described in Example 1. [Figure 2-2] FIG. 2 shows [ 18 F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-750142 from the NHP model described in Example 1. [Figure 2-3] FIG. 2 shows [ 18 F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-750142 from the NHP model described in Example 1. [Figure 3A] FIG. 3A shows the relationship between target occupancy and total NBI-750142 plasma concentrations in the NHP model described in Example 1. [Figure 3B]FIG. 3B shows the estimated VMAT2 target occupancy (% TO) for human doses of NBI-750142 from the NHP model described in Example 1. [Figure 4A] Figures 4A-4B show [18F]AV-133 SUV images averaged at baseline and 90-120 min after NBI-98782 at various doses in two different cynomolgus monkeys: A7701 (Figure 2A) and A7702 (Figure 2B). A dose-dependent decrease in striatal uptake was observed. [Figure 4B] Figures 4A-4B show [18F]AV-133 SUV images averaged at baseline and 90-120 min after NBI-98782 at various doses in two different cynomolgus monkeys: A7701 (Figure 2A) and A7702 (Figure 2B). A dose-dependent decrease in striatal uptake was observed. [Figure 5-1] FIG. 5 shows [18F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-98782 from the NHP model described in Example 2. [Figure 5-2] FIG. 5 shows [18F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-98782 from the NHP model described in Example 2. [Figure 5-3] FIG. 5 shows [18F]AV-133 time-activity curves comparing baseline and various dose levels of NBI-98782 from the NHP model described in Example 2. [Figure 6] FIG. 6 shows the relationship between target occupancy and total NBI-98782 plasma concentrations from the NHP model described in Example 2. [Figure 7A]Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 7B] Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 7C] Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 7D]Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 7E] Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 7F] Figures 7A-7F show [18F]AV-133 SUVR (occipital lobe reference region) images averaged over 60-120 min at baseline, and 1.5 h (T1) and 18 h (T2) after NBI-750142 administration in humans: Cohort 1: 100 mg (Figure 7A); Cohort 2: 200 mg (Figure 7B); Cohort 3: 60 mg (Figure 7C); Cohort 4: 60 mg (Figure 7D); Cohort 5A: 10 mg (Figure 7E); and Cohort 5B: 10 mg (Figure 7F). MRI images are shown for anatomical reference. [Figure 8A-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8A-2]8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8B-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8B-2] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8C-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8C-2]8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8D-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8D-2] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8E-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8E-2]8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8F-1] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 8F-2] 8A-8F show [18F]AV-133 SUV time-activity curves in humans at baseline and 1.5 hours (T1) and 18 hours (T2) after administration of NBI-750142: Cohort 1: 100 mg (Figure 8A); Cohort 2: 200 mg (Figure 8B); Cohort 3: 60 mg (Figure 8C); Cohort 4: 60 mg (Figure 8D); Cohort 5A: 10 mg (Figure 8E); and Cohort 5B: 10 mg (Figure 8F). [Figure 9A] 9A-9B show the relationship between NBI-750142 striatal occupancy and total plasma concentration based on the Emax model for both T1 and T2 (FIG. 9A), and T1 only (FIG. 9B). [Figure 9B] 9A-9B show the relationship between NBI-750142 striatal occupancy and total plasma concentration based on the Emax model for both T1 and T2 (FIG. 9A), and T1 only (FIG. 9B). [Figure 10] 10A-10B show a comparison of Emax and upregulation model fits for striatal occupancy at T1 and T2 (FIG. 10A) and T1 only (FIG. 10B) versus total plasma concentration across cohorts and subjects. [Figure 11]FIG. 11 shows the estimated VMAT2 target occupancy (% TO) for NBI-98782 from the NHP model described in Example 2. [Figure 12] Figure 12 shows the estimated VMAT2 occupancy of valbenazine over each 24-hour period. Figure 12 also shows cynomolgus monkey pharmacokinetic data with estimates of %TO at selected time points. As described below, adverse events were observed at these doses, allowing for the evaluation of %TO values ​​that may result in adverse events. [Figure 13] Figure 13A shows the estimated VMAT2 concentration-dependent target occupancy (%TO) for NBI-98782 (EC50 1.5ng / mL). Figure 13B shows the estimated VMAT2 concentration-dependent target occupancy (%TO) for NBI-98782 (EC50 3.8ng / mL). [Figure 14] FIG. 14 shows the imaging and sampling procedure for non-human primates. [Figure 15] FIG. 15 shows the protocol for estimating VMAT2 occupancy. [Figure 16] FIG. 16 shows the concentration versus time curves for [+]-α-HTBZ during a PET scan session. [Figure 17] FIG. 17 shows the percent target occupancy achieved in non-human primate studies. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] Detailed Description The present application relates to a method for preparing a pharmaceutical composition comprising a VMAT2 inhibitor, and / or a method for identifying a therapeutically effective dose of a VMAT2 inhibitor. In some embodiments, the VMAT2 inhibitors provided herein are particularly useful for achieving 80-96% occupancy in a subject, as described herein, and for treating neurological and psychiatric diseases and disorders. There is a significant unmet need for identifying a therapeutically effective dose of a VMAT2 inhibitor that provides 80-96% occupancy (e.g., for reducing or preventing treatment-emergent adverse events (TEAEs) associated with VMAT2 inhibition). The present disclosure meets these and other needs, as will be apparent with reference to the disclosure below.

[0009] In the following description, certain specific details are presented to provide a thorough understanding of various embodiments. However, those skilled in the art will understand that the present invention may be practiced without these details. In other instances, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless the context requires otherwise, throughout this specification and the claims that follow, the word "comprise" and variations thereof, such as "comprises" and "comprising," should be interpreted in an open and inclusive sense, i.e., "including but not limited to."

[0010] The headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed invention. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0011] Also, as used in this specification and the appended claims, the singular forms "a," "an," and "the" include the plural forms unless the content clearly dictates otherwise.

[0012] How to use Accordingly, the present application provides a method for preparing a pharmaceutical composition comprising a therapeutically effective dose of a VMAT2 inhibitor, comprising: (a) administering to a subject an amount of a VMAT2 inhibitor; (b) measuring in vivo VMAT2 occupancy of the VMAT2 inhibitor in the subject, wherein a VMAT2 occupancy of 80-96% indicates that the amount of the VMAT2 inhibitor is a therapeutically effective dose; and (c) combining a therapeutically effective dose of a VMAT2 inhibitor with a pharma- ceutically acceptable carrier. The present invention provides a method comprising:

[0013] The present application provides a method for identifying a therapeutically effective dose of a VMAT2 inhibitor, comprising: (a) administering to a subject an amount of a VMAT2 inhibitor; (b) measuring the in vivo VMAT2 occupancy of a VMAT2 inhibitor in a subject; and (c) identifying a therapeutically effective dose of the VMAT2 inhibitor when the amount of the VMAT2 inhibitor provides a VMAT2 occupancy rate of 80-96%; The present invention further provides a method comprising:

[0014] In some embodiments, VMAT2 occupancy is measured by one or more imaging techniques. In some embodiments, the one or more imaging techniques include administering to the subject an imaging agent that can bind to VMAT2, and then imaging the subject.

[0015] In some embodiments, the imaging agent is a VMAT2 inhibitor.

[0016] In some embodiments, VMAT2 occupancy is measured by positron emission tomography (PET) assay. In some embodiments, PET assay comprises administering to subject a PET imaging agent that can bind to VMAT2, and then imaging the subject.

[0017] In some embodiments, the PET assay comprises: (a) administering to a subject a PET imaging agent capable of binding to VMAT2; (b) waiting a sufficient time for the PET imaging agent to bind to VMAT2; (c) imaging the object one or more times; (d) measuring VMAT2 displacement of the PET imaging agent; and (e) determining VMAT2 occupancy based on the measured VMAT2 displacement of the PET imaging agent. Includes.

[0018] In some embodiments, VMAT2 displacement of the PET imaging agent is measured at one or more time points during imaging.

[0019] In some embodiments, the PET assay further comprises imaging the subject before step (a) to obtain a baseline image. In some embodiments, the VMAT2 inhibitor is administered to the subject after step (a). In some embodiments, the VMAT2 inhibitor is administered to the subject after step (b). In some embodiments, the VMAT2 inhibitor is administered to the subject after step (b) and before step (c).

[0020] In some embodiments, the PET imaging agent is a radiolabeled VMAT2 inhibitor. In some embodiments, the PET imaging agent is 11 In some embodiments, the PET imaging agent is a [C]-radiolabeled VMAT2 inhibitor. 18 F]-radiolabelled VMAT2 inhibitor.

[0021] In some embodiments, the PET imaging agent is a radiolabeled analog of a VMAT2 inhibitor selected from the group consisting of valbenazine, tetrabenazine, deutetrabenazine, dihydrotetrabenazine, NBI-750142, and AV-133.

[0022] In some embodiments, the PET imaging agent is a VMAT2 inhibitor selected from the group consisting of valbenazine, tetrabenazine, deutetrabenazine, dihydrotetrabenazine, NBI-750142, and AV-133. 11 C]- or [ 18 F]-radiolabelled analogue. In some embodiments, the radiolabeled analogue of dihydrotetrabenazine is a radiolabeled analogue of (+)-α-dihydrotetrabenazine.

[0023] In some embodiments, the PET imaging agent is 18 F]-AV-133.

[0024] In some embodiments, the method provided herein further comprises measuring the plasma concentration of the VMAT2 inhibitor in the subject.In some embodiments, the plasma concentration of the VMAT2 inhibitor is measured at one or more time points during the imaging of step (c).In some embodiments, the plasma concentration of the VMAT2 inhibitor is measured at one or more time points before the imaging of step (c).In some embodiments, the plasma concentration of the VMAT2 inhibitor is measured at one or more time points during the imaging of step (c) and before the imaging of step (c).

[0025] In some embodiments, the plasma concentration of the VMAT2 inhibitor is measured at one or more time points starting from about 2 hours prior to the imaging of step (c) to the end of imaging.

[0026] In some embodiments, the plasma concentration of the VMAT2 inhibitor is measured at one or more time points starting from about 1 hour prior to the imaging of step (c) to the end of imaging.

[0027] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% to about 96%, e.g., about 80%, about 82%, about 84%, about 86%, about 88%, about 90%, about 92%, about 94%, or about 96%.

[0028] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not greater than 96%.

[0029] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not greater than 94%.

[0030] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not greater than 92%.

[0031] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not greater than 90%.

[0032] In some embodiments, the methods provided herein further include monitoring the subject for one or more symptoms associated with treatment-emergent adverse events (TEAEs) after administration of the VMAT2 inhibitor.

[0033] In some embodiments, monitoring is performed for about 30 minutes to about 24 hours after administration of the VMAT2 inhibitor, for example, about 30 minutes, about 60 minutes, about 90 minutes, about 120 minutes, about 180 minutes, about 6 hours, about 12 hours, about 18 hours, or about 24 hours.

[0034] In some embodiments, the monitoring occurs for about 30 minutes to about 90 minutes after administration of the VMAT2 inhibitor.

[0035] In some embodiments, the monitoring occurs for about 30 minutes to about 60 minutes after administration of the VMAT2 inhibitor.

[0036] In some embodiments, the methods provided herein further include identifying the subject as not exhibiting one or more symptoms associated with a TEAE following administration of the VMAT2 inhibitor.

[0037] In some embodiments, the methods provided herein further include identifying the subject as not exhibiting one or more symptoms selected from ptosis, hypoactivity, sedation, anxiety, nausea, akathisia, and salivation after administration of the VMAT2 inhibitor.

[0038] In some embodiments, the methods provided herein further include identifying the subject as not exhibiting one or more symptoms selected from ptosis, hypoactivity, and drooling after administration of the VMAT2 inhibitor.

[0039] In some embodiments, the subject is identified as not exhibiting one or more symptoms associated with a TEAE following administration of the VMAT2 inhibitor.

[0040] In some embodiments, the subject is identified as not exhibiting one or more symptoms selected from ptosis, hypoactivity, sedation, anxiety, nausea, akathisia, and salivation after administration of the VMAT2 inhibitor.

[0041] In some embodiments, the subject is identified as not exhibiting one or more symptoms selected from ptosis, hypoactivity, and drooling following administration of the VMAT2 inhibitor.

[0042] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not greater than 96%, e.g., about 80%, about 82%, about 84%, about 86%, about 88%, about 90%, about 92%, about 94%, or about 96%.

[0043] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 85% and not greater than 95%.

[0044] In some embodiments, an administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 85% and not greater than 90%.

[0045] In some embodiments, the method comprises: (i) Radioactive ligand [ 11 C](+) 4-Propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol ([ 11 C]-PHNO) to a subject; and (ii) Image scanning of the object. and optionally measuring by Non-substitutable bonds ([ 11 C]-PHNO BP ND ) compared to [ 11 A 20-45% increase in [C]-PHNO binding capacity corresponds to a decrease in synaptic dopamine, indicating that the amount of VMAT2 inhibitor is a therapeutically effective dose.

[0046] The present application provides a method for preparing a pharmaceutical composition comprising a therapeutically effective dose of a VMAT2 inhibitor, comprising: mixing a therapeutically effective dose of a VMAT2 inhibitor with a pharma- ceutically acceptable carrier; Including, The therapeutically effective dose of a VMAT2 inhibitor is Measuring the in vivo VMAT2 occupancy of a VMAT2 inhibitor in a subject previously administered an amount of the VMAT2 inhibitor, where a VMAT2 occupancy rate of 80-96% indicates that the amount of the VMAT2 inhibitor is a therapeutically effective dose. The present invention further provides a method, characterized by:

[0047] In some embodiments, the method comprises: (i) Radioactive ligand [ 11 C](+) 4-Propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol ([ 11 C]-PHNO) to a subject; and (ii) Image scanning of the object. and optionally measuring the Non-substitutable bonds ([ 11 C]-PHNO BP ND ) compared to [ 11 A 20–45% increase in [C]-PHNO binding capacity corresponded to a decrease in synaptic dopamine, indicating that the amount of VMAT2 inhibitor administered was therapeutically effective.

[0048] In some embodiments, [ 11 C]-PNHO BP ND The administered dose of VMAT2 inhibitor is identified as a therapeutically effective dose when the increase in is determined to be at least 10% to about 60%, e.g., about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55% or about 60%. In such an embodiment, 11 C]-PNHO BP ND An increase in β-dopa corresponds to a decrease in synaptic dopamine.

[0049] The term "compound" (e.g., VMAT2 inhibitor) as used herein is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the depicted structure. Compounds identified herein by name or structure as one particular tautomeric form are intended to include other tautomeric forms unless otherwise specified.

[0050] As used herein, "VMAT2" refers to human vesicular monoamine transporter isoform 2, an integral membrane protein that acts to transport monoamines, particularly neurotransmitters such as dopamine, norepinephrine, serotonin and histamine, from the cell cytosol into synaptic vesicles.

[0051] As used herein, the term "VMAT2 inhibitor", "inhibit VMAT2", or "inhibit VMAT2" refers to the ability of the compounds disclosed herein to alter the function of VMAT2. VMAT2 inhibitors can block or reduce the activity of VMAT2 by forming a reversible or irreversible covalent bond between the inhibitor and VMAT2, or through the formation of a non-covalently bound complex. Such inhibition may only be manifested in certain cell types or may be associated with certain biological events. The term "VMAT2 inhibitor", "inhibit VMAT2", or "inhibit VMAT2" also refers to altering the function of VMAT2 by reducing the probability of complex formation between VMAT2 and a natural substrate.

[0052] A substance is an "inhibitor" of enzyme activity if it can reduce the specific activity of an enzyme or the metabolic effect of specific activity by its presence, regardless of the exact mechanism of such reduction.For example, a substance can be an inhibitor of enzyme activity by competitive, non-competitive, allosteric or other types of enzyme inhibition, by reducing the expression of the enzyme, or by other direct or indirect mechanisms.The co-administration of a given drug with an inhibitor can reduce the rate of metabolism of the drug through the metabolic pathways listed.

[0053] By way of example only, in some embodiments, "synaptic dopamine" can be measured non-invasively as follows. 11 Oral administration of a VMAT2 inhibitor was performed before administration of [C]-PHNO, and PET imaging was performed before and after the peak plasma concentration of the VMAT2 inhibitor. ND ) was used as the primary endpoint. Using the cerebellum as the reference region, regional BP ND Estimate the change in [ 11 C]-PHNO BP ND Increase in (ΔBP ND) corresponds to a decrease in synaptic dopamine after administration of a VMAT2 inhibitor. Other radioligands can also be used. Alternatively, in some embodiments, indirect measurement of synaptic dopamine can be achieved via invasive techniques, such as those described in Huang M., et al., Pharmacology, Biochemistry and Behavior 190 (2020) 172872; and Bosche SL, et al., Frontiers in Neuroscience, November 2021 | Volume 15 | Article 728092.

[0054] "Image scanning" of a subject refers to subjecting a subject to a scan, for example, a positron emission tomography (PET) and / or computed tomography (CT) scan.

[0055] The present application also includes pharma- ceutically acceptable salts of the compounds described herein (e.g., pharma- ceutically acceptable salts of the VMAT2 inhibitors described herein). As used herein, "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds, in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues, e.g., amines; alkali or organic salts of acidic residues, e.g., carboxylic acids, and the like. The pharma- ceutically acceptable salts of the present disclosure include conventional non-toxic salts of the parent compound, for example, formed from non-toxic inorganic or organic acids. The pharma- ceutically acceptable salts of the present disclosure can be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water, in an organic solvent, or in a mixture of the two, generally with non-aqueous media such as ether, ethyl acetate, alcohol (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (ACN) being preferred. Lists of suitable salts can be found in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., 1985, p. 1418 and Journal of Pharmaceutical Science, 66, 2 (1977), each of which is incorporated herein by reference in its entirety.

[0056] As will be appreciated by those of skill in the art, the compounds provided herein, including their salts and stereoisomers, can be prepared using known organic synthesis techniques, or can be synthesized according to any of a number of possible synthetic routes.

[0057] As used herein, "valbenazine" may be referred to as (S)-2-amino-3-methyl-butyric acid (2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1a]isoquinolin-2-yl ester; or L-valine, (2R,3R,11bR)-1,3,4,6,7,11b-hexahydro-9,10-dimethoxy-3-(2-methylpropyl)-2H-benzo[a]quinolizin-2-yl ester, or NBI-98854.

[0058] As used herein, "NBI-98782" or "(+)-α-HTBZ" has the structure: [ka] It refers to a compound that is an active metabolite of valbenazine, having the formula: (+)-α-HTBZ may also be referred to as (2R,3R,11bR), or as (+)-α-DHTBZ, or as (+)-α-HTBZ, or as R,R,R-DHTBZ, or as (+)-α-3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ol; or as (2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ol.

[0059] As used herein, the term "NBI-750142" refers to a compound having the following structure: [ka]

[0033] refers to a compound having the formula: See, e.g., U.S. Patent No. 11,040,970, the disclosure of which is incorporated herein by reference in its entirety.

[0060] The compounds (e.g., VMAT2 inhibitors) and pharmaceutical compositions (e.g., pharmaceutical compositions comprising VMAT2 inhibitors) described herein can inhibit the activity of VMAT2 in subjects.Compounds that inhibit VMAT2 are useful for providing a means of treating neurological and psychiatric diseases and disorders.

[0061] Exemplary neurological and psychiatric diseases and disorders associated with VMAT2 include, but are not limited to, hyperkinetic disorder (i.e., hyperkinetic movement disorder or "hyperkinesia").Further examples of diseases or disorders that can be treated by VMAT2 inhibition can be found in, for example, U.S. Patent Nos. 10,065,952, 10,857,137, 10,857,148, 10,912,771, 10,952,997, 10,993,941, 11,026,931 and 11,040,029; the disclosures of which are incorporated herein by reference in their entirety; and in U.S. Patent Application Nos. 20200078352 and 20200397779; the disclosures of which are incorporated herein by reference in their entirety.

[0062] As used herein, "hyperkinetic disorder" or "hyperkinetic movement disorder" or "hyperkinesia" refers to a disorder or disease characterized by excessive, abnormal, involuntary movements. These neurological disorders include tremor, dystonia, myoclonus, athetosis, Huntington's disease, tardive dyskinesia, Tourette's syndrome, dystonia, hemiballismus, chorea, senile chorea, or tics.

[0063] As used herein, "tardive syndrome" includes, but is not limited to, tardive dyskinesia, tardive dystonia, tardive akathisia, tardive tics, myoclonus, tremor, and withdrawal-emergent syndrome. Tardive dyskinesia is characterized by rapid, repetitive, stereotyped, involuntary movements of the face, limbs, or trunk.

[0064] As used herein, "administering" refers to the process of introducing a composition or dosage form into a patient via art-recognized means of introduction.

[0065] As used herein, the term "disorder" is intended to be generally synonymous with, and is used interchangeably with, the terms "disease," "syndrome," and "condition" (as in medical condition), in that all reflect an abnormal condition of the human or animal body or one of its parts that impairs normal functioning and is typically manifested by characteristic signs and symptoms.

[0066] As used herein, "dose" refers to the measured quantity of active agent taken by a patient at one time. In certain embodiments where the active agent is not the free base form of the VMAT2 inhibitor, the amount is the molar equivalent to the equivalent amount of the free base form of the VMAT2 inhibitor. For example, drugs are often packaged in pharma- ceutically acceptable salt form, and dosages in terms of strength refer to the mass of the molar equivalent of the corresponding free base. As used herein, "dose" administered is used interchangeably with "amount" administered.

[0067] As used herein, an "effective amount" and a "therapeutically effective amount" of an agent, compound, drug, composition or combination is an amount that is non-toxic and effective to produce some desired therapeutic effect upon administration to a subject or patient (e.g., a human subject or patient). The exact therapeutically effective amount for a subject may depend, for example, on the subject's size and health, the nature and extent of the condition, the therapeutic agent or combination of therapeutic agents selected for administration, and other variables known to those skilled in the art. The effective amount for a given situation is determined by routine experimentation and is within the judgment of the clinician.

[0068] As used herein, "patient" or "subject" means a mammal, including a human, for whom treatment is desired, and generally refers to the recipient of the treatment.

[0069] As used herein, "pharmacologically acceptable" refers to a material that is not biologically or otherwise undesirable, i.e., the material can be incorporated into a pharmaceutical composition administered to a patient without causing undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. When the term "pharmacologically acceptable" is used to refer to a pharmaceutical carrier or excipient, it means that the carrier or excipient has met the necessary standards of toxicological and manufacturing testing, or that the carrier or excipient is included in the Inactive Ingredients Guide prepared by the U.S. Food and Drug Administration. "Pharmacologically active" (or simply "active"), as in "pharmacologically active" (or "active") derivative or analog, refers to a derivative or analog that has the same type of pharmacological activity as the parent compound, and in roughly the same degree. The term "pharmacologically acceptable salt" includes acid addition salts formed with inorganic acids, such as hydrochloric acid or phosphoric acid, or organic acids, such as acetic acid, oxalic acid, tartaric acid, mandelic acid, and the like. Salts formed with the free carboxyl groups can also be derived from inorganic bases such as, for example, sodium, potassium, ammonium, calcium, or ferric hydroxides, and such organic bases as isopropylamine, trimethylamine, histidine, procaine, and the like.

[0070] As used herein, "treating" or "treatment" refers to therapeutic applications to slow or stop the progression of a disorder, prophylactic applications to prevent the onset of a disorder, and / or reversal of a disorder. Reversal of a disorder differs from therapeutic applications that slow or stop a disorder in that the method of reversal not only completely stops the progression of the disorder, but also causes the behavior of cells to move to some degree toward the normal state that would be observed in the absence of the disorder.

[0071] Pharmaceutical Compositions Also provided herein is a pharmaceutical composition for use in treating a neurological or psychiatric disease or disorder, comprising a VMAT2 inhibitor as an active pharmaceutical ingredient in combination with one or more pharma- ceutically acceptable carriers or excipients.

[0072] The choice of excipient will to a large extent depend on factors such as the particular mode of administration, the effect of the excipient on the solubility and stability of the active ingredient, and the nature of the dosage form.

[0073] The pharmaceutical compositions provided herein may be provided in unit dosage form or multiple dosage form. Unit dosage form, as used herein, refers to a physically separate unit suitable for administration to human and animal subjects and packaged individually as known in the art. Each unit dose contains a predetermined amount of active ingredient(s) sufficient to produce the desired therapeutic effect, together with the required pharmaceutical carrier or excipient. Examples of unit dosage forms include ampoules, syringes, and individually packaged tablets and capsules. A unit dosage form may be administered in portions or multiples thereof. A multiple dosage form is a plurality of identical unit dosage forms packaged in a single container to be administered in separate unit dosage form. Examples of multiple dosage forms include vials, bottles of tablets or capsules, or bottles of pints or gallons.

[0074] The pharmaceutical compositions provided herein may be administered alone or in combination with one or more other compounds provided herein, one or more other active ingredients. The pharmaceutical compositions provided herein may be formulated in various dosage forms for oral, parenteral and topical administration. The pharmaceutical compositions may also be formulated as modified release dosage forms, including delayed release, extended release, prolonged release, sustained release, pulsed release, controlled release, accelerated and rapid release, targeted release, programmed release, and gastric retention dosage forms. These dosage forms may be prepared according to conventional methods and techniques known to those skilled in the art. The pharmaceutical compositions provided herein may be administered once or multiple times at intervals of time. It is understood that the exact dosage and duration of treatment may vary according to the age, weight and condition of the patient being treated, and may be determined empirically using known testing protocols or by extrapolation from in vivo or in vitro test or diagnostic data. It is further understood that for any particular individual, the specific dosage regimen should be adjusted over time according to the individual's needs and the professional judgment of the person administering or monitoring the administration of the formulation.

[0075] Oral route The pharmaceutical compositions provided herein may be provided in solid, semi-solid or liquid dosage forms for oral administration. As used herein, oral administration also includes buccal, lingual and sublingual administration. Suitable oral dosage forms include, but are not limited to, tablets, capsules, pills, troches, lozenges, pastilles, cachets, pellets, medicated chewing gums, granules, bulk powders, effervescent or non-effervescent powders or granules, solutions, emulsions, suspensions, solutions, wafers, sprinkles, elixirs and syrups. In addition to the active ingredient(s), pharmaceutical compositions may contain one or more pharma- ceutically acceptable carriers or excipients, including, but not limited to, binders, fillers, diluents, disintegrants, wetting agents, lubricants, glidants, colorants, dye transfer inhibitors, sweeteners and flavoring agents.

[0076] Binders or granulating agents impart cohesiveness to the tablet to ensure that the tablet remains intact after compression. Suitable binders or granulators include starches such as corn starch, potato starch and pregelatinized starch (e.g., STARCH 1500); gelatin; sugars such as sucrose, glucose, dextrose, molasses and lactose; natural and synthetic gums such as acacia, alginic acid, alginates, Irish moss extract, panwar gum, ghatti gum, mucilage of isabgol shell, carboxymethylcellulose, methylcellulose, polyvinylpyrrolidone (PVP), Veegum, larch arabinogalactan, powdered tragacanth and guar gum; celluloses such as ethyl cellulose, cellulose acetate, calcium carboxymethylcellulose, sodium carboxymethylcellulose, methylcellulose, hydroxyethylcellulose (HEC), hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC); microcrystalline celluloses such as AVICEL-PH-101, AVICEL-PH-103, AVICEL-PH-104, AVICEL-PH-105, AVICEL-PH-106, AVICEL-PH-107, AVICEL-PH-108, AVICEL-PH-109, AVICEL-PH-200, AVICEL-PH-201, AVICEL-PH-202, AVICEL-PH-203, AVICEL-PH-204, AVICEL-PH-205, AVICEL-PH-206, AVICEL-PH-207, AVICEL-PH-208, AVICEL-PH-209, AVICEL-PH-210, AVICEL-PH-211, AVICEL-PH-212, AVICEL-PH-213, AVICEL-PH-214, AVICEL-PH-215, AVICEL-PH-216, AVICEL-PH-217, AVICEL-PH-218, AVICEL-PH-219, AVICEL-PH Suitable fillers include, but are not limited to, RC-581, AVICEL-PH-105 (FMC Corp., Marcus Hook, PA); and mixtures thereof. Suitable fillers include, but are not limited to, talc, calcium carbonate, microcrystalline cellulose, powdered cellulose, dextrates, kaolin, mannitol, silicic acid, sorbitol, starch, pregelatinized starch, and mixtures thereof. The binder or filler may be present in the pharmaceutical compositions provided herein at about 50 to about 99% by weight.

[0077] Suitable diluents include, but are not limited to, dicalcium phosphate, calcium sulfate, lactose, sorbitol, sucrose, inositol, cellulose, kaolin, mannitol, sodium chloride, dry starch and powdered sugar.Certain diluents, such as mannitol, lactose, sorbitol, sucrose and inositol, when present in sufficient quantities, can impart to some compressed tablets the property of allowing them to disintegrate in the mouth by chewing.Such compressed tablets can be used as chewable tablets.

[0078] Suitable disintegrants include, but are not limited to, agar; bentonite; cellulose, such as methylcellulose and carboxymethylcellulose; wood products; natural sponge; cation exchange resins; alginic acid; gums, such as guar gum and veegum HV; citrus pulp; cross-linked cellulose, such as croscarmellose; cross-linked polymers, such as crospovidone; cross-linked starch; calcium carbonate; microcrystalline cellulose, such as sodium starch glycolate; polacrilin potassium; starches, such as com starch, potato starch, tapioca starch, and pregelatinized starch; clay; align; and mixtures thereof. The amount of disintegrant in the pharmaceutical compositions provided herein varies with the type of formulation and is readily discernible to one of skill in the art. The pharmaceutical compositions provided herein may contain about 0.5 to about 15% by weight or about 1 to about 5% by weight of disintegrant.

[0079] Suitable lubricants include, but are not limited to, calcium stearate, magnesium stearate, mineral oil, light mineral oil, glycerin, sorbitol, mannitol, glycols, such as glycerol behenate and polyethylene glycol (PEG), stearic acid, sodium lauryl sulfate, talc, hydrogenated vegetable oils, including peanut oil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, and soybean oil, zinc stearate, ethyl oleate, ethyl laurate, agar, starch, watercress, silica or silica gel, such as AEROSIL® 200 (WR Grace Co., Baltimore, MD) and CAB-0-SIL® (Cabot Co., Boston, MA), and mixtures thereof. The pharmaceutical compositions provided herein may contain about 0.1 to about 5% by weight of a lubricant. Suitable glidants include colloidal silicon dioxide, CAB-0-SIL® (Cabot Co., Boston, MA) and asbestos-free talc. Coloring agents include any of the approved, certified water-soluble FD and C dyes and water-insoluble FD and C dyes and lakes suspended on alumina hydrate, and mixtures thereof. Lakes are combinations of water-soluble dyes adsorbed onto hydrous oxides of heavy metals to produce an insoluble form of the dye. Flavoring agents include natural flavors extracted from plants such as fruits, and synthetic blends of compounds that produce a pleasant taste, such as peppermint and methyl salicylate. Sweetening agents include sucrose, lactose, mannitol, syrups, glycerin, and artificial sweeteners such as saccharin and aspartame. Suitable emulsifying agents include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate (TWEEN® 20), polyoxyethylene sorbitan monooleate 80 (TWEEN® 80), and triethanolamine oleate.Suspending and dispersing agents include sodium carboxymethylcellulose, pectin, tragacanth, veegum, acacia, sodium carbomethylcellulose, hydroxypropylmethylcellulose and polyvinylpyrrolidone. Preservatives include glycerin, methyl and propylparaben, benzoic add, sodium benzoate and alcohol. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauryl ether. Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examples of non-aqueous liquids utilized in emulsions include mineral oil and cottonseed oil. Organic acids include citric acid and tartaric acid. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate.

[0080] It should be understood that many carriers and excipients may perform multiple functions even within the same formulation. The pharmaceutical compositions provided herein may be provided as compressed tablets, tablet triturates, chewable lozenges, fast dissolving tablets, multiple compressed tablets, or enteric coated, sugar-coated or film-coated tablets. Enteric coated tablets are compressed tablets coated with a substance that resists the action of stomach acid but dissolves or disintegrates in the intestine, thereby protecting the active ingredients from the acidic environment of the stomach. Enteric coatings include, but are not limited to, fatty acids, fats, phenyl salicylates, waxes, shellac, ammoniated shellac, and cellulose acetate phthalates. Sugar-coated tablets are compressed tablets surrounded by a sugar coating, which may be beneficial in masking unpleasant tastes or odors and protecting the tablets from oxidation. Film-coated tablets are compressed tablets covered with a thin layer or film of a water-soluble material. Film coatings include, but are not limited to, hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000, and cellulose acetate phthalates. Film coatings impart the same general characteristics as sugar coatings. Multiple compressed tablets are compressed tablets made by multiple compression cycles, and include layered tablets, and press-coated or dry-coated tablets.

[0081] Tablet dosage forms can be prepared from the active ingredient in powdered, crystalline or granular form, alone or in combination with one or more carriers or excipients described herein, including binders, disintegrants, controlled release polymers, lubricants, diluents and / or colorants. Flavoring and sweetening agents are particularly useful in the formation of chewable tablets and lozenges.

[0082] The pharmaceutical compositions provided herein may be provided as soft or hard capsules, which may be made from gelatin, methylcellulose, starch, or calcium alginate. Hard gelatin capsules, also known as dry-filled capsules (DFC), consist of two sections, one nested over the other, thus completely enclosing the active ingredient. Soft elastic capsules (SEC) are soft, globular shells, such as gelatin shells, plasticized by the addition of glycerin, sorbitol, or similar polyols. Soft gelatin shells may contain preservatives to prevent microbial growth. Suitable preservatives are those described herein, including methyl and propyl parabens and sorbic acid. The liquid, semi-solid, and solid dosage forms provided herein may be encapsulated in capsules. Suitable liquid and semi-solid dosage forms include solutions and suspensions in propylene carbonate, vegetable oils, or triglycerides. Capsules may also be coated, as known to those skilled in the art, to modify or prolong the dissolution of the active ingredient.

[0083] The pharmaceutical compositions provided herein may be provided in liquid and semisolid dosage forms, including emulsions, solutions, suspensions, elixirs, and syrups. Emulsions are biphasic systems in which one liquid is dispersed in another in the form of small globules, and may be oil-in-water or water-in-oil. Emulsions may contain a pharma-ceutically acceptable non-aqueous liquid or solvent, an emulsifier, and a preservative. Suspensions may contain a pharma-ceutically acceptable suspending agent and a preservative. Aqueous alcoholic solutions may contain pharma-ceutically acceptable acetals, such as di(lower alkyl)acetals of lower alkyl aldehydes (the term "lower" means alkyl having 1 to 6 carbon atoms), such as acetaldehyde diethyl acetal; and water-miscible solvents having one or more hydroxyl groups, such as propylene glycol and ethanol. Elixirs are clear, sweetened aqueous alcoholic solutions. Syrups are concentrated aqueous solutions of sugars, such as sucrose, and may also contain preservatives. For a liquid dosage form, the solution, for example, in a polyethylene glycol, may be diluted with a sufficient quantity of a pharma- ceutically acceptable liquid carrier, for example water, to be conveniently measured for administration.

[0084] Other useful liquid and semisolid dosage forms include, but are not limited to, those containing the active ingredient(s) provided herein and dialkylated mono- or polyalkylene glycols, including 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether, where 350, 550, and 750 refer to the approximate average molecular weight of the polyethylene glycol. These formulations may further include one or more antioxidants, such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarin, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, bisulfites, sodium metabisulfite, thiodipropionic acid and its esters, and dithiocarbamates.

[0085] The pharmaceutical compositions provided herein for oral administration may also be provided in the form of liposomes, micelles, microspheres, or nanosystems.

[0086] The pharmaceutical compositions provided herein may be provided as non-effervescent or effervescent granules and powders that are reconstituted into liquid dosage forms. Pharmaceutically acceptable carriers and excipients used in non-effervescent granules or powders may include diluents, sweeteners, and wetting agents. Pharmaceutically acceptable carriers and excipients used in effervescent granules or powders may include organic acids and sources of carbon dioxide. Colorants and flavoring agents may be used in all of the above dosage forms. The pharmaceutical compositions provided herein may be formulated as immediate release dosage forms or modified release dosage forms, including delayed release, sustained release, pulsed release, controlled release, targeted release, and programmed release forms.

[0087] The pharmaceutical compositions provided herein may be co-formulated with other active ingredients that do not impair the desired therapeutic action, or with substances that supplement the desired action, such as antacids, proton pump inhibitors, and H2 receptor antagonists.

[0088] The pharmaceutical compositions provided herein can be administered parenterally by injection, infusion, or implantation for local or systemic administration. As used herein, parenteral administration includes intravenous, intraarterial, intraperitoneal, intrathecal, intravenous, intraurethral, ​​intrasternal, intracranial, intramuscular, intrasynovial, and subcutaneous administration.

[0089] Parenteral Administration The pharmaceutical compositions provided herein can be formulated into any dosage form suitable for parenteral administration, including liquids, suspensions, emulsions, micelles, liposomes, microspheres, nanosystems, and solid forms suitable for solution or suspension in liquid prior to injection. Such dosage forms can be prepared according to conventional methods known to those skilled in the art of pharmaceutical science.

[0090] Pharmaceutical compositions intended for parenteral administration may contain one or more pharma- ceutically acceptable carriers and excipients including, but not limited to, aqueous vehicles, water-miscible vehicles, non-aqueous vehicles, antimicrobial agents or preservatives against microbial growth, stabilizers, solubility enhancers, isotonicity agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, cryoprotectants, lyophilization protectants, thickening agents, pH adjusting agents, and inert gases.

[0091] Suitable aqueous vehicles include, but are not limited to, water, saline, normal saline or phosphate buffered saline (PBS), sodium chloride injection, Ringer's injection, isotonic dextrose injection, sterile water injection, dextrose and lactated Ringer's injection.Non-aqueous vehicles include, but are not limited to, fixed oils of vegetable origin, castor oil, corn oil, cottonseed oil, olive oil, peanut oil, peppermint oil, safflower oil, sesame oil, soybean oil, hydrogenated vegetable oil, hydrogenated soybean oil, and medium-chain triglycerides of coconut oil and palm kernel oil.Water-miscible vehicles include, but are not limited to, ethanol, 1,3-butanediol, liquid polyethylene glycols (e.g., polyethylene glycol 300 and polyethylene glycol 400), propylene glycol, glycerin, N-methyl-2-pyrrolidone, dimethylacetamide, and dimethylsulfoxide.

[0092] Suitable antimicrobial or preservative agents include, but are not limited to, phenol, cresol, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl phydroxybenzate, thimerosal, benzalkonium chloride, benzethonium chloride, methyl and propyl paraben, and sorbic acid. Suitable isotonicity agents include, but are not limited to, sodium chloride, glycerin, and dextrose. Suitable buffers include, but are not limited to, phosphates and citrates. Suitable antioxidants are those described herein, including bisulfites and sodium metabisulfite. Suitable local anesthetics include, but are not limited to, procaine hydrochloride. Suitable suspending and dispersing agents are those described herein, including sodium carboxymethylcellulose, hydroxypropylmethylcellulose, and polyvinylpyrrolidone. Suitable emulsifying agents include those described herein, including polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate 80, and triethanolamine oleate. Suitable sequestering or chelating agents include, but are not limited to, EDTA. Suitable pH adjusting agents include, but are not limited to, sodium hydroxide, hydrochloric acid, citric acid, and lactic acid. Suitable complexing agents include, but are not limited to, cyclodextrins, including alpha-cyclodextrin, beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin, sulfobutylether-beta-cyclodextrin, and sulfobutylether 7-beta-cyclodextrin (CAPTISOL®, CyDex, Lenexa, KS).

[0093] The pharmaceutical compositions provided herein can be formulated for administration by single dose or multiple doses. Single dose formulations are packaged in ampoules, vials or syringes. Multiple dose parenteral formulations must contain antibacterial agents at bacteriostatic or fungistatic concentrations. As known and practiced in the art, all parenteral formulations must be sterile.

[0094] In certain embodiments, the pharmaceutical composition is provided as a ready-to-use sterile liquid. In certain embodiments, the pharmaceutical composition is provided as a sterile dry soluble product, including lyophilized powder and hypodermic tablets, which are reconstituted with a vehicle before use. In certain embodiments, the pharmaceutical composition is provided as a ready-to-use sterile suspension. In certain embodiments, the pharmaceutical composition is provided as a sterile dry insoluble product, which is reconstituted with a vehicle before use. In certain embodiments, the pharmaceutical composition is provided as a ready-to-use sterile emulsion.

[0095] The pharmaceutical compositions provided herein may be formulated as immediate or modified release dosage forms, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release forms.

[0096] The pharmaceutical compositions may be formulated as a suspension, solid, semi-solid, or thixotropic liquid for administration as an implanted depot. In certain embodiments, the pharmaceutical compositions provided herein are dispersed in an inner solid matrix surrounded by an outer polymeric membrane that is insoluble in body fluids but permits diffusion of the active ingredients of the pharmaceutical composition.

[0097] Suitable inner matrices include polymethyl methacrylate, polybutyl methacrylate, plasticized or unplasticized polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of esters of acrylic and methacrylic acid, collagen, crosslinked polyvinyl alcohol, and crosslinked partially hydrolyzed polyvinyl acetate.

[0098] Suitable outer polymeric membranes include polyethylene, polypropylene, ethylene / propylene copolymers, ethylene / ethyl acrylate copolymers, ethylene / vinyl acetate copolymers, silicone rubber, polydimethylsiloxane, neoprene rubber, chlorinated polyethylene, polyvinyl chloride, vinyl acetate, vinylidene chloride, vinyl chloride copolymers with ethylene and propylene, ionomeric polyethylene terephthalate, butyl rubber, epichlorohydrin rubber, ethylene / vinyl alcohol copolymers, ethylene / vinyl acetate / vinyl alcohol terpolymers, and ethylene / vinyloxyethanol copolymers.

[0099] Topical administration The pharmaceutical compositions provided herein can be administered topically to the skin, orifices, or mucous membranes.As used herein, topical administration includes intradermal, intraconjuctival, intracorneal, intraocular, ophthalmic, auricular, transdermal, nasal, vaginal, urethral, ​​respiratory, and rectal administration.

[0100] The pharmaceutical compositions provided herein may be formulated into any form suitable for topical administration for local or systemic effect, including emulsions, solutions, suspensions, creams, gels, hydrogels, ointments, dusting powders, coatings, elixirs, lotions, suspensions, tinctures, pastes, foams, films, aerosols, irrigations, sprays, suppositories, bandages, skin patches, etc. Topical formulations of the pharmaceutical compositions provided herein may also include liposomes, micelles, microspheres, nanosystems, and mixtures thereof.

[0101] Pharmaceutically acceptable carriers and excipients suitable for use in the topical formulations provided herein include, but are not limited to, aqueous vehicles, water-miscible vehicles, non-aqueous vehicles, antimicrobial agents or preservatives against microbial growth, stabilizers, solubility enhancers, isotonicity agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, wetting or emulsifying agents, complexing agents, sequestering or chelating agents, penetration enhancers, cryoprotectants, lyoprotectants, viscosity enhancing agents, and inert gases.

[0102] Pharmaceutical compositions may also be administered locally by electroporation, iontophoresis, phonophoresis, sonophoresis, and microneedle or needle-free injection, for example, POWDERJECT™ (Chiron Corp., Emeryville, Calif.) and BIOJECT™ (Bioject Medical Technologies Inc., Tualatin, Oreg.).

[0103] The pharmaceutical composition provided herein may be provided in the form of ointment, cream and gel.Suitable ointment vehicles include oily or hydrocarbon bases, including lard, benzoic lard, olive oil, cottonseed oil and other oils, white petrolatum, etc.; emulsifying or absorbing bases, such as hydrophilic petrolatum, hydroxystearin sulfate, and anhydrous lanolin; water-removing bases, such as hydrophilic ointments; water-soluble ointment bases, including polyethylene glycols of various molecular weights; emulsion bases, either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, including cetyl alcohol, glyceryl monostearate, lanolin, and stearic acid.These vehicles are emollient, but generally require the addition of antioxidants and preservatives.

[0104] Suitable cream bases may be oil-in-water or water-in-oil. Cream vehicles may be water-washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase is also called the "internal" phase and is generally composed of petrolatum and a fatty alcohol, such as cetyl alcohol or stearyl alcohol. The aqueous phase usually, but not necessarily, exceeds the oil phase in volume and generally contains a moisturizer. The emulsifier in cream formulations may be a nonionic surfactant, an anionic surfactant, a cationic surfactant, or an amphoteric surfactant.

[0105] Gels are semi-solid suspension-type systems. Single-phase gels contain organic polymers that are distributed substantially uniformly throughout the liquid carrier. Suitable gelling agents include cross-linked acrylic acid polymers, such as carbomers, carboxypolyalkylenes, Carbopol®; hydrophilic polymers, such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers, and polyvinyl alcohol; cellulosic polymers, such as hydroxypropylcellulose, hydroxyethylcellulose, hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate, and methylcellulose; gums, such as tragacanth and xanthan gum; sodium alginate; and gelatin. To prepare a uniform gel, dispersing agents, such as alcohol or glycerin, may be added, or the gelling agent may be dispersed by trituration, mechanical mixing, and / or stirring.

[0106] The pharmaceutical compositions provided herein may be administered rectally, urethrally, vaginally, or perivaginally in the form of a suppository, pessary, bougie, poultice or cataplasm, paste, powder, coating, cream, plaster, contraceptive, ointment, solution, emulsion, suspension, tampon, gel, foam, spray, or enema. These dosage forms can be manufactured using conventional processes.

[0107] Rectal, urethral, ​​and vaginal suppositories are solid objects for insertion into body openings, solid at normal temperatures, but melt or soften at body temperature to release the active ingredient(s) inside the opening. Pharmaceutically acceptable carriers utilized for rectal and vaginal suppositories include vehicles such as stiffening agents that provide a melting point close to body temperature when formulated with the pharmaceutical compositions provided herein; and antioxidants as described herein, including bisulfite and sodium metabisulfite. Suitable vehicles include, but are not limited to, cocoa butter (theobroma oil), glycerin-gelatin, carbowax (polyoxyethylene glycol), spermaceti, paraffin, white and yellow wax, and suitable mixtures of mono-, di-, and triglycerides of fatty acids, hydrogels such as polyvinyl alcohol, hydroxyethyl methacrylate, polyacrylic acid; glycerin gelatin. Combinations of various vehicles can be used. Rectal and vaginal suppositories can be prepared by compression methods or molding. The typical weight of a rectal and vaginal suppository is about 2-3 g.

[0108] The pharmaceutical compositions provided herein can be administered ophthalmically in the form of solutions, suspensions, ointments, emulsions, gel-forming solutions, powders / sprinkles for solutions, gels, ocular inserts, and implants.

[0109] The pharmaceutical composition provided herein can be administered intranasally or by inhalation to the airway.The pharmaceutical composition can be provided in the form of an aerosol or liquid for delivery using a pressurized container, pump, spray, atomizer, such as an atomizer that uses electrohydrodynamics to generate a fine mist, or a nebulizer, alone or in combination with a suitable propellant, such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane.The pharmaceutical composition can also be provided as a dry powder for insufflation, and as a nasal drop, alone or in combination with an inert carrier, such as lactose or phospholipid.For intranasal use, the powder can include a bioadhesive agent, including chitosan or cyclodextrin.

[0110] Solutions or suspensions for use in pressurized containers, pumps, sprays, atomizers, or nebulizers can be formulated to contain ethanol, aqueous ethanol, or a suitable substitute, a propellant as a solvent; and / or a surfactant, e.g., sorbitan trioleate, oleic acid, or oligolactic acid, for dispersing, solubilizing, or extending the release of the active ingredients provided herein.

[0111] The pharmaceutical compositions provided herein can be micronized to a size suitable for delivery by inhalation, for example, 50 micrometers or less, or 10 micrometers or less.Particles of such size can be prepared using comminution methods known to those skilled in the art, for example, spiral jet milling, fluidized bed jet milling, supercritical fluid processing to form nanoparticles, high pressure homogenization, or spray drying.

[0112] Capsules, blisters and cartridges for use in an inhaler or insufflator can be formulated to contain a powder mix of the pharmaceutical composition provided herein; a suitable powder base, such as lactose or starch; and performance modifiers, such as / -leucine, mannitol, or magnesium stearate. Lactose can be anhydrous or in the form of monohydrate. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose. The pharmaceutical composition provided herein for inhalation / intranasal administration can further contain suitable flavors, such as menthol and levomenthol, or sweeteners, such as saccharin or saccharin sodium.

[0113] The pharmaceutical compositions provided herein for topical administration may be formulated to be immediate or modified release, including delayed-, sustained-, pulsed-, controlled-, targeted-, and programmed-release.

[0114] controlled release The pharmaceutical compositions provided herein can be formulated as modified release dosage forms.As used herein, the term "modified release" refers to dosage forms in which the rate or location of active ingredient release is different from that of immediate release dosage forms when administered by the same route.Modified release dosage forms include delayed release dosage forms, extended release dosage forms, prolonged release dosage forms, sustained release dosage forms, pulsatile or pulsed release dosage forms, controlled release dosage forms, accelerated and fast release dosage forms, targeted release dosage forms, program release dosage forms, and gastric retention dosage forms.

[0115] Pharmaceutical compositions in modified release dosage forms can be prepared using a variety of modified release devices and methods known to those skilled in the art, including, but not limited to, matrix controlled release devices, osmotic controlled release devices, multiparticulate controlled release devices, ion exchange resins, enteric coatings, multilayer coatings, microspheres, liposomes, and combinations thereof. The release rate of the active ingredient(s) can also be modified by varying the particle size and polymorphism of the active ingredient(s).

[0116] The pharmaceutical compositions provided herein in modified release dosage forms can be prepared using matrix controlled release devices known to those of ordinary skill in the art.

[0117] In certain embodiments, the pharmaceutical compositions provided herein in modified release dosage forms are formulated using erodible matrix devices that are water-swellable, erodible, or soluble polymers, including synthetic polymers, and natural polymers and derivatives, such as polysaccharides and proteins.

[0118] Materials useful for forming the erodible matrix include chitin, chitosan, dextran and pullulan; agar gum, gum arabic, karaya gum, locust bean gum, tragacanth gum, carrageenan, ghatti gum, guar gum, xanthan gum, and scleroglucan; starches, such as dextrin and maltodextrin; hydrocolloids, such as pectin; phosphatides, such as lecithin; alginates; propylene glycol alginate; gelatin; collagen; and cellulose derivatives, such as ethyl cellulose (EC), methyl ethyl cellulose (MEC), carboxymethyl cellulose (CMC), CMEC, hydroxyethyl cellulose (HCMC), cellulose esters, ... cellulose (HEC), hydroxypropyl cellulose (HPC), cellulose acetate (CA), cellulose propionate (CP), cellulose butyrate (CB), cellulose acetate butyrate (CAB), CAP, CAT, hydroxypropyl methylcellulose (HPMC), HPMCP, HPMCAS, hydroxypropyl methylcellulose acetate trimellitate (HPMCAT) and ethyl hydroxyethyl cellulose (EHEC); polyvinylpyrrolidone; polyvinyl alcohol; polyvinyl acetate; glycerol fatty acid esters; polyacrylamide; polyacrylic acid; copolymers of ethacrylic or methacrylic acid (EUDRAGIT®, Rohm copolymers of L-glutamic acid and ethyl-L-glutamate; degradable lactic acid-glycolic acid copolymers; poly-D-(-)-3-hydroxybutyric acid; and other acrylic acid derivatives, such as, but not limited to, homopolymers and copolymers of butyl methacrylate, methyl methacrylate, ethyl methacrylate, ethyl acrylate, (2-dimethylaminoethyl) methacrylate, and (trimethylaminoethyl) methacrylate chloride.

[0119] In certain embodiments, the pharmaceutical compositions are formulated with a non-erodible matrix device: the active ingredient is dissolved or dispersed in an inert matrix and, once administered, is released primarily by diffusion through the inert matrix. Materials suitable for use as non-erodible matrix devices include insoluble plastics such as polyethylene, polypropylene, polyisoprene, polyisobutylene, polybutadiene, polymethyl methacrylate, polybutyl methacrylate, chlorinated polyethylene, polyvinyl chloride, methyl acrylate-methyl methacrylate copolymers, ethylene-vinyl acetate copolymers, ethylene / propylene copolymers, ethylene / ethyl acrylate copolymers, vinyl acetate, vinylidene chloride, copolymers of vinyl chloride with ethylene and propylene, ionomeric polyethylene terephthalate, butyl rubber, epichlorohydrin rubber, ethylene / vinyl alcohol copolymers, ethylene / vinyl acetate / vinyl alcohol terpolymers, and ethylene / vinyloxyethanol copolymers, polyvinyl chloride, plasticized nylon, plasticized polyethylene terephthalate, natural rubber, silicone rubber, polydimethylsiloxane, silicone carbonate copolymers, and hydrophilic polymers such as ethyl cellulose, cellulose acetate, crospovidone, and crosslinked partially hydrolyzed polyvinyl acetate; and aliphatic compounds such as carnauba wax. Examples of suitable waxes include, but are not limited to, glyceryl stearate ...

[0120] In a matrix controlled release system, the desired release kinetics can be controlled, for example, through the type of polymer used, the viscosity of the polymer, the particle size of the polymer and / or the active ingredient(s), the ratio of active ingredient(s) to polymer, and other excipients in the composition.

[0121] The pharmaceutical compositions provided herein in modified release dosage forms can be prepared by methods known to those skilled in the art, including direct compression, dry or wet granulation followed by compression, and melt granulation followed by compression.

[0122] The pharmaceutical composition provided herein in a modified release dosage form can be manufactured using an osmotic controlled release device, including one-container system, two-container system, asymmetric membrane technology (AMT) and extruded core system (ECS).In general, such a device has at least two components: (a) a core that contains active ingredient(s); and (b) a semipermeable membrane that encapsulates the core and has at least one delivery port.The semipermeable membrane controls the influx of water from the aqueous environment of use into the core, so that the drug is released by extrusion through delivery port(s).

[0123] In addition to the active ingredient(s), the core of the osmotic device optionally contains an osmotic agent that creates a driving force for transporting water from the environment of use into the core of the device. One class of osmotic agents are water-swellable hydrophilic polymers, which are also referred to as "osmopolymers" and "hydrogels." Suitable osmotic agents include, but are not limited to, hydrophilic vinyl and acrylic polymers, polysaccharides such as calcium alginate, polyethylene oxide (PEO), polyethylene glycol (PEG), polypropylene glycol (PPG), poly(2-hydroxyethyl methacrylate), poly(acrylic) acid, poly(methacrylic) acid, polyvinylpyrrolidone (PVP), crosslinked PVP, polyvinyl alcohol (PVA), PVA / PVP copolymers, PVA / PVP copolymers with hydrophobic monomers such as methyl methacrylate and vinyl acetate, hydrophilic polyurethanes containing large PEO blocks, croscarmellose sodium, carrageenan, hydroxyethyl cellulose (HEC), hydroxypropyl cellulose (HPC), hydroxypropyl methylcellulose (HPMC), carboxymethyl cellulose (CMC), and carboxyethyl cellulose (CEC), sodium alginate, polycarbophil, gelatin, xanthan gum, and sodium starch glycolate.

[0124] Another class of osmotic agent is osmogen, which can absorb water and affect the osmotic pressure gradient across the barrier of the surrounding coating.Suitable osmogens include, but are not limited to, inorganic salts such as magnesium sulfate, magnesium chloride, calcium chloride, sodium chloride, lithium chloride, potassium sulfate, potassium phosphate, sodium carbonate, sodium sulfite, lithium sulfate, potassium chloride and sodium sulfate; sugars such as dextrose, fructose, glucose, inositol, lactose, maltose, mannitol, raffinose, sorbitol, sucrose, trehalose and xylitol; organic acids such as ascorbic acid, benzoic acid, fumaric acid, citric acid, maleic acid, sebacic acid, sorbic acid, adipic acid, edetic acid, glutamic acid, p-toluenesulfonic acid, succinic acid and tartaric acid; urea; and mixtures thereof.

[0125] Osmotic agents with different dissolution rates can be used to affect the rate at which active ingredient(s) are initially delivered from the dosage form.For example, amorphous sugars such as Mannogeme EZ (SPI Pharma, Lewes, DE) can be used to provide a more rapid delivery during the first few hours, so as to rapidly produce the desired therapeutic effect, and the remaining amount is gradually and continuously released to maintain the desired level of therapeutic or prophylactic effect over a long period of time.In this case, active ingredient(s) is released at a rate that replaces the amount of active ingredient that is metabolized and excreted.

[0126] The core may also contain a wide variety of other excipients and carriers as described herein to enhance the performance of the dosage form or to promote stability or processing.

[0127] Materials useful for forming the semipermeable membrane include various grades of acrylics, vinyls, ethers, polyamides, polyesters, and cellulosics that are water permeable and water insoluble at physiologically relevant pH or susceptible to being rendered water insoluble by chemical changes such as crosslinking. Examples of suitable polymers useful for forming the coating include plasticized, unplasticized, and reinforced cellulose acetate (CA), cellulose diacetate, cellulose triacetate, CA propionate, cellulose nitrate, cellulose acetate butyrate (CAB), CA ethyl carbamate, CAP, CA methyl carbamate, CA succinate, cellulose acetate trimellitate (CAT), CA dimethylaminoacetate, CA ethyl carbonate, CA chloroacetate, CA ethyl oxalate, CA methyl sulfonate, CA butyl sulfonate, CA p-toluenesulfonate, agar acetate, amylose triacetate, beta glucan acetate, beta glucan triacetate, acetaldehyde dimethyl acetate, triacetate of locust bean gum, hydroxide ethylene vinyl acetate, EC, PEG, PPG, PEG / PPG copolymers, PVP, HEC, HPC, CMC, CMEC, HPMC, HPMCP, HPMCAS, HPMCAT, poly(acrylic) acids and esters, and poly(methacrylic) acids and esters and copolymers thereof, starch, dextran, dextrin, chitosan, collagen, gelatin, polyalkenes, polyethers, polysulfones, polyethersulfones, polystyrene, polyvinyl halides, polyvinyl esters and ethers, natural and synthetic waxes.

[0128] The semipermeable membrane may also be a hydrophobic microporous membrane, as disclosed in US Patent No. 5,798,119, in which the pores are substantially filled with gas and are not wetted by aqueous media but are water permeable. Such hydrophobic but water permeable membranes are typically composed of hydrophobic polymers, such as polyalkenes, polyethylene, polypropylene, polytetrafluoroethylene, polyacrylic acid derivatives, polyethers, polysulfones, polyethersulfones, polystyrene, polyvinyl halides, polyvinylidene fluorides, polyvinyl esters and ethers, natural waxes, and synthetic waxes. The delivery port(s) on the semipermeable membrane can be formed after coating by mechanical or laser drilling. The delivery port(s) can also be formed in situ by erosion of a plug of water-soluble material or by rupture of a thinner portion of the membrane above the core recess. In addition, the delivery port may be formed during the coating process.

[0129] The total amount and release rate of the active ingredient(s) released can be substantially modulated via the thickness and porosity of the semipermeable membrane, the composition of the core, and the number, size and location of the delivery ports.

[0130] The pharmaceutical composition in an osmotic controlled release dosage form may further comprise additional conventional excipients as described herein to facilitate performance or processing of the formulation.

[0131] The osmotic controlled release dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art.

[0132] In certain embodiments, the pharmaceutical compositions provided herein are formulated as AMT controlled release dosage forms, which comprise an asymmetric osmotic membrane coating a core comprising an active ingredient(s) and other pharma-ceutical acceptable excipients. AMT controlled release dosage forms can be prepared according to conventional methods and techniques known to those skilled in the art, including direct compression, dry granulation, wet granulation, and dip-coating methods.

[0133] In certain embodiments, the pharmaceutical compositions provided herein are formulated as ESC controlled release dosage forms comprising an osmotic membrane coating a core comprising the active ingredient(s), hydroxyethylcellulose, and other pharma- ceutically acceptable excipients.

[0134] The pharmaceutical compositions provided herein in modified release dosage forms can be manufactured as multiparticulate controlled release devices comprising a multiplicity of particles, granules or pellets having diameters ranging from about 10 pm to about 3 mm, about 50 pm to about 2.5 mm, or about 100 pm to about 1 mm. Such multiparticulates can be made by processes known to those skilled in the art, including wet or dry granulation, extrusion / spheronization, roller compaction, melt congealing, and by spray coating seed cores.

[0135] Other excipients as described herein can be blended with the pharmaceutical composition to aid in the processing and formation of the multiparticulates. The resulting particles may themselves constitute the multiparticulate device or may be coated with various film-forming materials, such as enteric polymers, water-swellable, and water-soluble polymers. The multiparticulates can be further processed into capsules or tablets.

[0136] targeted delivery The pharmaceutical compositions provided herein may also be formulated to target specific tissues, receptors, or other areas of the body of the subject receiving the treatment, and include liposome-based, resealed erythrocyte-based, and antibody-based delivery systems.

[0137] labeled compound In some embodiments, the VMAT2 inhibitors provided herein are isotopic variants of VMAT2 inhibitors. As used herein, "isotopic variant" refers to a compound that contains an unnatural proportion of isotopes in one or more of the atoms that constitute such compound. In certain embodiments, the "isotopic variant" of a compound is an isotope of hydrogen ( 1 H), deuterium ( 2 H), tritium ( 3 H), Carbon-11( 11 C), carbon-12( 12 C), carbon-13( 13 C), carbon-14( 14 C), nitrogen-13( 13 N), nitrogen-14( 14 N), nitrogen-15( 15 N), oxygen-14( 14 O), oxygen-15( 15 O), oxygen-16( 16 O), oxygen-17( 17 O), oxygen-18( 18 O), Fluorine-17( 17 F), Fluorine-18( 18 F), Phosphorus-31( 31 P), phosphorus-32( 32 P), phosphorus-33( 33 P), sulfur-32( 32 S), Sulfur-33( 33 S), Sulfur-34( 34 S), Sulfur-35( 35 S), Sulfur-36( 36 S), Chlorine-35( 35 Cl), Chlorine-36( 36 Cl), Chlorine-37( 37 Cl), Bromine-79( 79 Br), Bromine-81( 81 Br), Iodine-123( 123 I), iodine-125( 125 I), iodine-127( 127 I), iodine-129( 129 I), and iodine-131( 131In certain embodiments, an "isotopic variant" of a compound is in a stable form, i.e., is non-radioactive. In certain embodiments, an "isotopic variant" of a compound is in a stable form, i.e., is non-radioactive. In certain embodiments, an "isotopic variant" of a compound is in a stable form, i.e., is non-radioactive. 1 H), deuterium ( 2 H), Carbon-12( 12 C), carbon-13( 13 C), nitrogen-14( 14 N), nitrogen-15( 15 N), oxygen-16( 16 O), oxygen-17( 17 O), and oxygen-18 ( 18 In certain embodiments, an "isotopic variant" of a compound is in an unstable form, i.e., is radioactive. In certain embodiments, an "isotopic variant" of a compound is in an unstable form, i.e., is radioactive. In certain embodiments, an "isotopic variant" of a compound is in an unstable form, i.e., is radioactive. 3 H), Carbon-11( 11 C), carbon-14( 14 C), nitrogen-13( 13 N), oxygen-14( 14 O), and oxygen-15 ( 15 In the compounds provided herein, any hydrogen may be, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 109, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121 2 H or any carbon can be, for example 13 C or any nitrogen, e.g. 15 N, any oxygen, e.g. 18 It is understood that the isotope ratio can be O. In certain embodiments, an "isotopic variant" of a compound contains unnatural proportions of deuterium.

[0138] With respect to the compounds provided herein, when a particular atomic position is designated as having deuterium or "D" or "d", it is understood that the abundance of deuterium at that position is substantially greater than the natural abundance of deuterium, which is about 0.015%. Positions designated as having deuterium typically have a minimum isotopic enrichment factor at each designated deuterium position of, in certain embodiments, at least 1000 (15% deuterium incorporation), at least 2000 (30% deuterium incorporation), at least 3000 (45% deuterium incorporation), at least 3500 (52.5% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation). The isotopic enrichment of the compounds provided herein can be determined using conventional analytical methods known to those of skill in the art, including mass spectrometry, nuclear magnetic resonance spectroscopy, and crystallography.

[0139] In some embodiments, the present application provides radiolabeled compounds, and uses thereof. In some embodiments, the radiolabeled compounds provided herein include 11 C. 13 N, 15 O. 18 F, 68 Ga, 89 Zr, 82 Rb, 124 I, and 131 The radioisotope comprises at least one radioisotope selected from the group consisting of I. In some embodiments, the radioisotope is a positron emitter. As used herein, the term "positron emitter" refers to a radioisotope in which protons are converted to neutrons, thereby emitting a positron and an electron neutrino. In some embodiments, the positron emitter is 11 C or 18F. In some embodiments, the radiolabeled compounds provided herein comprise at least one 18 F radioisotope. In some embodiments, the radiolabeled compounds provided herein are suitable for use as imaging agents. In some embodiments, the radiolabeled compounds provided herein are suitable for use as positron emission tomography (PET) imaging agents.

[0140] kit The present application further includes pharmaceutical kits, for example, useful in the treatment of diseases and disorders mentioned herein, which include one or more containers containing the pharmaceutical compositions described herein. Such kits can further include one or more of a variety of conventional pharmaceutical kit components, such as a container with one or more pharma- ceutically acceptable carriers, additional containers, etc., as would be readily apparent to one skilled in the art. Instructions, either as inserts or labels, indicating the amounts of components to be administered, guidelines for administration, and / or guidelines for mixing the components, can also be included in the kit.

[0141] Abbreviation The following abbreviations may be used throughout this application: [Table A] EXAMPLES

[0142] The following examples are provided to better illustrate the claimed invention and should not be construed as limiting the scope of the invention. To the extent that specific materials are mentioned, they are merely for illustrative purposes and are not intended to limit the invention. Those skilled in the art may develop equivalent means or reactants without the exercise of inventive capacity and without departing from the scope of the invention.

[0143] Example 1 [ 18VMAT2 receptor occupancy study in non-human primates (NHPs) after intravenous (iv) administration of NBI-98782 using [F]AV-133 positron emission tomography (PET) The purpose of this study is to evaluate the dose / plasma concentration of NBI-98782 and its association with [ 18 The aim of this study was to evaluate the relationship between VMAT2 occupancy and the degree of VMAT2 occupancy induced by [F]AV-133. NBI-98782 (i.e., [+]-α-HTBZ; or (+)-α-3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ol; or (2R,3R,11bR)-3-isobutyl-9,10-dimethoxy-1,3,4,6,7,11b-hexahydro-2H-pyrido[2,1-a]isoquinolin-2-ol) is an active metabolite of valbenazine that has been developed as a therapeutic agent to inhibit VMAT2 and decrease dopamine release at presynaptic nerve terminals. The radioligand [ 18 [F]AV-133 has been used to image and quantify brain VMAT2 in vivo. PET studies in NHPs have been shown to provide useful information on brain access and target engagement by novel compounds. Such data can be used to demonstrate target engagement of novel CNS compounds to index pharmacodynamic outcomes, which, for example, allows for the identification of dose ranges with important implications in clinical practice.

[0144] [ 18 F]AV-133 Labeling Protocol [ 18 [F]AV-133 was prepared from the following radiosynthetic scheme: [ka] High performance liquid chromatography (HPLC) chromatograms of the final purified radiolabeled [ 18 The purity of [F]AV-133 was demonstrated.

[0145] Preparation of dosage forms [ 18 [F]AV-133 was formulated in saline containing ethanol and ascorbic acid. NBI-98782 was dissolved in 0.9% sterile saline and the resulting solution was filtered through a 0.2 μm sterile filter into an empty sterile glass vial. To avoid possible losses during filtration, approximately 5 mL of the formulated material was filtered and discarded before collection of the final dose formulation. The solution was visually inspected and found to be clear and free of particles. For all preparations, concentrations are reported as the free base of NBI-98782 using a correction factor of 1.73.

[0146] Animal models Two female non-naive cynomolgus monkeys (Macaca fascicularis) were used in this study. The animals were judged to be in good health prior to the start of the study. At the start of the study, animals A7701 (tag ID A5028) and A7702 (tag ID A5029) weighed 4.45 kg / 4 years 11 months and 4.75 kg / 3 years 10 months, respectively.

[0147] In vivo imaging studies Animals were treated with vehicle (0.9% saline) at baseline. 18 F]AV-133 and on a separate day after NBI-98782 blockade. Vehicle and NBI-98782 were administered as an intravenous (iv) bolus followed by continuous infusion over 3 hours. One hour after vehicle / NBI98782 administration, [ 18 [F]AV-133 was injected as an intravenous bolus. The study design is summarized in Table 1-1. [Table 1-1]

[0148] Dose Administration On the day of dose administration, each animal was weighed. Two intravenous lines were placed to allow for (1) administration of the radiopharmaceutical ([ 18F]AV-133) and (2) administration of test article (NBI-98782) or vehicle (0.9% saline). See Tables 1-2 and 1-3 for details.

[0149] Monkeys were fasted overnight prior to each individual PET scan. Animals were sedated with an intramuscular injection of ketamine 5–10 mg / kg prior to administration of NBI-98782 or vehicle. Animals were intubated with an endotracheal tube for continuous delivery of isoflurane to achieve maintenance anesthesia. Hydration was maintained by intravenous administration of lactated Ringer's solution (LRS) at 3–5 mL / kg / h. Levels of isoflurane and fluids were adjusted over the course of the study to maintain anesthesia.

[0150] Body temperature was maintained using a warm water blanket.Vital signs, including heart rate, respiratory rate, oxygen saturation, and temperature, were monitored at least every 10-15 min throughout the period the monkeys were under anesthesia.

[0151] Image acquisition and processing Dynamic data were acquired with a MicroPET Focus-220 camera (Siemens Microsystems, Knoxville, TN). Luminescence data were collected for 120 min post-injection and reconstructed into a sequence of 33 frames, applying all standard corrections: normalization, random, scattering and attenuation (via CT scan). Dynamic brain PET images were transferred and analyzed using the image processing PMOD software package v3.802 (PMOD Technologies, Zurich, Switzerland). PET images were rigorously registered to a previously acquired T1 MRI of the animal's brain and subsequently spatially normalized to a common cynomolgus monkey MRI template for definition of anatomical brain regions.

[0152] Analysis method Evaluation of NBI-98782 concentration During each scan, the following time points were recorded: pre-administration (-5 min), 4 min after administration of the test substance (approximately 1 min after the bolus was completed), 30 min, and 60 min after administration ([ 18Whole blood PK samples (in K2 EDTA tubes) were collected at 90, 120, 150, and 180 minutes (just before administration of [F]AV-133 radiopharmaceutical), 90, 120, 150, and 180 minutes (end of scan). PK samples were approximately 1 mL each (8 mL total including pre-dose sample). PK sample collection times were relative to the start of NBI-98782 bolus administration.

[0153] NBI-98782 concentrations in cynomolgus monkey plasma samples were analyzed using a validated LC-MS / MS method. The quantitative range of the NBI-98782 assay is 0.50-500 ng / mL, with dilution integrity demonstrated up to 12,500 ng / mL. Plasma samples were analyzed using stable isotope-labeled internal standards. 13 The samples were processed using liquid-liquid extraction with the addition of C-83198. The processed samples were chromatographically separated on a C18 reversed phase column and interfaced to a Sciex API 4000 triple quadrupole mass spectrometer for detection. The average concentrations of NBI-98782 are presented in Tables 1-6.

[0154] Non-GLP Determination of NBI-98782 in Stabilized K2-EDTA Plasma of Cynomolgus Monkeys in a Cyno PET Imaging Study by LC-MS-MS The method was validated for NBI-98782 in the range of 0.500–500 ng / mL based on the analysis of 100 μL of cynomolgus monkey plasma by LC-MS-MS. Quantitation was performed using a weighted 1 / x2 linear least-squares regression analysis generated from calibration standards prepared on the day of extraction. Liquid-liquid extraction was performed using an internal standard solution (approximately 400 ng / mL [ 13 C]-Radioactive label[ 13 C]-NBI-98854 (i.e., [ 13 10.0 μL of [C]-valbenazine) and [ 13The incubation was started by the addition of [C]-NBI-83198 (racemic mixture of two dihydrotetrabenazine enantiomers). All samples received 50.0 μL of sodium carbonate solution (0.2 M) and 2.00 mL of methyl tert-butyl ether, after which the tubes were sealed, vortexed, and centrifuged. The organic layer was then transferred to a clean tube and evaporated to dryness. The samples were then reconstituted with 150 μL of reconstitution solution (water / methanol / acetic acid; 80:20:0.2), sealed, vortexed, sonicated, and centrifuged. The reconstituted extracts were then transferred to a 96-well plate. Chromatographic separation was achieved using an HPLC Kinetex C18 column (2.6 μm, 50 × 2.1 mm) with gradient elution using mobile phase A (water / ammonium acetate solution / acetic acid; 1000:5:1) and mobile phase B (2-propanol / methanol / ammonium acetate solution / acetic acid; 300:700:5:1). A triple quadrupole mass spectrometer (Sciex API 4000) equipped with a TurboIonSpray source was used in positive ion mode. Quantitation was performed using NBI-98782 and [ 13 This was based on multiple reaction monitoring (MRM) of the m / z transitions of [C]-NBI-83198.

[0155] Visualization of tissue uptake A volume of interest (VOI) defined within the cynomolgus monkey template space (see, for example, Ballanger et al. Neuroimage, 2013, 77:26-43) was applied to the spatially normalized PET images to define the caudate nucleus (0.96 cm 3 ), putamen (1.34cm 3 ) and occipital lobe (8.61 cm 3 ) for time-activity curve (TAC, kBq / cm 3 ) was calculated. TAC and images (averaged over 90–120 min) are presented in standard uptake value (SUV) units (g / mL) normalized by animal weight and injected dose.

[0156] Image quantification Non-invasive Logan graph analysis (start time of fit, t* = 30 min) was used to estimate regional non-displaceable binding potential (BPND) using the cerebellar cortex as the reference TAC and the caudate and putamen TACs. BPND was calculated in the caudate and putamen across baseline and NBI-98782 scans. Target occupancy was calculated according to Equation 2-1.

number

number

[0157] result Two female cynomolgus monkeys were randomly assigned to receive NBI-98782 at baseline and after administration of NBI-98782. 18 The relationship between the mean total NBI-98782 concentration during the scan (mean concentration for 60-180 min after administration of NBI-98782) and the VMAT2 occupancy in the striatum (averaged from the caudate and putamen) was examined using [F]AV-133 (5.5 ± 0.9 mCi). Tables 1-2 and 1-3 show the relationship between the mean total NBI-98782 concentration during the scan (mean concentration for 60-180 min after administration of NBI-98782) and the VMAT2 occupancy in the striatum (averaged from the caudate and putamen). 18 F]AV-133 and NBI-98782 injection information is shown. Throughout the evaluation, 18 [F]AV-133 SUV images are shown in Figure 4 and TAC in Figure 5.

[0158] At baseline, the caudate and putamen 18 Uptake of [F]AV-133 was higher in the sham-treated rats than in the rest of the brain. 18A dose- and concentration-dependent decrease in [F]AV-133 binding was observed following administration of NBI-98782. BPND and occupancy are shown in Tables 1-4 for animal A7701 and 1-5 for animal A7702. Total NBI-98782 concentrations and [ 18 The striatal occupancy of [F]AV-133 is shown in Tables 1-6. The relationship between the occupancy of NBI-98782 and the total plasma concentration is shown in Figure 6. The 50% effective total concentration (EC 50 ) was 3.8 ng / mL, with a 95% confidence interval of 1.9 ng / mL to 7.5 ng / mL. [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6]

[0159] conclusion [ 18 NBI-98782 striatal occupancy of VMAT2 was concentration-dependent as measured by [F]AV-133 PET imaging. E using a Hill slope of 1.0 and a fixed maximum occupancy of 100% max Using the model, the EC 50 The free (unbound) EC was 3.8 ng / mL (95% confidence interval: 1.9 ng / mL to 7.5 ng / mL). 50 was 1.5 ng / mL (see, e.g., FIG. 13).

[0160] Using two valbenazine doses (40 mg and 80 mg) that demonstrated efficacy in a Phase 3 study for tardive dyskinesia, together with the known pharmacokinetic properties of valbenazine, the PET method described above was applied to estimate the VMAT2 target occupancy (%TO) at therapeutic levels of valbenazine for the treatment of TD. Steady-state free plasma concentrations of NBI-98782 in humans were estimated from total plasma concentrations of NBI-98782 and human PPB data after once-daily administration of 40 mg, 60 mg, or 80 mg of valbenazine. Free ECs obtained from NHP PET were then used to estimate the VMAT2 target occupancy (%TO) at therapeutic levels of valbenazine for the treatment of TD. 50 Using the maximal, mean and minimal plasma concentrations (C) of NBI-98782 at steady state based on a clinically effective dose of valbenazine, max , C ave , C trough ) was estimated for %TO in humans.

[0161] EC 50 Application of the data to steady-state human PK data demonstrated that once-daily administration of valbenazine maintained VMAT2 target occupancy as follows: 40 mg (73%-82%); 60 mg (82%-88%); 80 mg (85%-91%), as shown in FIG. 11. At doses demonstrated to be clinically effective in treating tardive dyskinesia, valbenazine is estimated to maintain high VMAT2 occupancy (≧73%) throughout each 24-hour period (see FIG. 11). Reasonable target occupancy (C ave Approximately 80% of patients with TD achieved VMAT2 occupancy at 40 mg valbenazine once daily, the recommended starting dose (and minimum approved dose) for all patients with TD, indicating potential biological efficacy from the start of treatment. At an estimated VMAT2 target occupancy of 96%-98% in cynomolgus monkeys (see Figure 12), clinical observations were recorded approximately 30 minutes to 1 hour after dosing, including partial eye closure (ptosis) (all monkeys), reduced activity (2 / 3 monkeys), and salivation (1 monkey).

[0162] Based on the larger effect size observed for valbenazine 80 mg compared to valbenazine 40 mg in the Phase 3 clinical trial, the results of this study suggest that high sustained occupancy (85% or higher) and VMAT2 inhibition support maximum efficacy against TD. Given that adverse effects in monkeys were observed at occupancy levels above 95%, these results suggest that once-daily administration of valbenazine 80 mg may be pharmacologically optimized to maximize efficacy (e.g., by achieving and maintaining above 85% TO) while avoiding adverse effects (by keeping TO generally below 95%). The process described herein matched the clinical efficacy of valbenazine with %TO estimates. These values, as summarized in Figure 12, may then serve as a reference value for comparison against any tetrabenazine analog VMAT2 inhibitor. Although these reference values ​​are defined by efficacy in treating TD, it is proposed that these reference values ​​will serve as a suitable "reference case" for exploring the efficacy of VMAT2 inhibitors in any therapeutic area.

[0163] Example 2 [ 18 VMAT2 receptor occupancy study in non-human primates (NHPs) after intravenous (iv) administration of NBI-750142 using [F]AV-133 positron emission tomography (PET) The purpose of this study is to evaluate the dose / plasma concentration of NBI-750142 and its association with [ 18 The aim of this study was to evaluate the relationship between VMAT2 occupancy and the effect of the radioligand [F]AV-133. 18[F]AV-133 has been used to image and quantify brain VMAT2 in vivo. PET studies in NHPs have been shown to provide useful information on brain access and target engagement by novel compounds. Such data can be used to demonstrate target engagement of novel CNS compounds to index pharmacodynamic outcomes, which, for example, allows for the identification of dose ranges with important implications in clinical practice.

[0164] [ 18 F]AV-133 Labeling Protocol [ 18 [F]AV-133 was prepared according to the following radiosynthetic scheme: [ka] High performance liquid chromatography (HPLC) chromatograms of the final purified radiolabeled [ 18 The purity of [F]AV-133 was demonstrated.

[0165] Preparation of dosage forms [ 18 [F]AV-133 was formulated in saline containing sodium ascorbate (4.66 mg / mL) and up to 10% ethanol. NBI-750142 was dissolved in 0.9% USP grade saline and the resulting solution was filtered through a 0.2 μm sterile filter into a sterile, empty glass vial. The solution was visually inspected and found to be clear and free of visible particles. For all preparations, concentrations are reported as the free base of NBI-750142 using a correction factor of 1.59.

[0166] Animal models Three non-naive cynomolgus monkeys (Macaca fascicularis) were used in this study. The animals were judged to be in good health prior to the start of the study. At the start of the study, the animals were 7 years old (female, EC865), 8 years old (male, GC786), and 9 years old (female, LC206), weighing 4.2 kg, 7.0 kg, and 5.4 kg, respectively.

[0167] In vivo imaging studies Animals were treated with vehicle (0.9% saline) at baseline. 18 F]AV-133 and on a separate day after NBI-750142 blockade. Vehicle and NBI-750142 were administered as an intravenous (iv) bolus followed by continuous infusion over 3 hours. One hour after vehicle / NBI-750142 administration, [ 18 [F]AV-133 was injected as an intravenous bolus. The study design is summarized in Table 2-1 below. [Table 2-1]

[0168] Dose Administration On the day of dose administration, each animal was weighed. Two intravenous lines were placed to allow for (1) administration of the radiopharmaceutical ([ 18 F]AV-133) and (2) administration of test article (NBI-750142) or vehicle (0.9% saline). See Table 2-2 for details. [Table 2-2]

[0169] Prior to each individual PET scan, monkeys were fasted for 8–12 h. At least 60 min before administration of NBI-750142 or vehicle, animals were sedated with a combination of Aflaxan 2 mg / kg, dexmedetomidine 0.02 mg / kg, and midazolam 0.3 mg / kg (intramuscular, IM) and transported to the imaging suite. One to two drops of 1% lidocaine were applied topically over the larynx to control inflammation induced by intubation. Animals were immediately intubated with an endotracheal tube and continuously delivered oxygen (1.5–2.5 L) and 1.0–2.5% isoflurane for anesthesia maintenance via a rebreathing or non-rebreathing circuit. Zofran 1.0 mg / kg (subcutaneous, SC) and dexamethasone 0.5 mg / kg (IM) were administered immediately after intubation to control nausea and inflammation, respectively. Hydration was maintained with lactated Ringer's solution (LRS) + 5% dextrose at 4-10 mL / kg / h (iv). Isoflurane and fluid levels were adjusted during the course of the study to maintain anesthesia.

[0170] Body temperature was maintained at 35–40°C using a warm water blanket. Vital signs, including heart rate, blood pressure, respiratory rate, oxygen saturation, and temperature, were monitored at least every 10–15 min throughout the period the monkeys were under anesthesia. Glycopyrrolate (0.01 mg / kg IM) was administered at the end of the study to control intubation-induced salivation.

[0171] Image acquisition and processing Dynamic data were acquired with a MicroPET Focus-220 camera (Siemens Microsystems, Knoxville, TN). Luminescence data was collected for 120 min post-injection and reconstructed into a sequence of 33 frames, with all standard corrections applied: normalization, random, scattering, and attenuation. Dynamic brain PET images were transferred and analyzed using the image processing PMOD software package v3.802 (PMOD Technologies, Zurich, Switzerland). PET images were rigorously registered to a previously acquired animal brain T1 MRI and subsequently spatially normalized to a common cynomolgus monkey MRI template for definition of anatomical brain regions.

[0172] Analysis method Evaluation of NBI-750142 concentration During each scan, the subjects were examined before administration (-5 min), 1 min after administration of the test substance (end of bolus), 30 min, and 60 min after administration ([ 18 Whole blood PK samples were collected (in K2 EDTA tubes) at 90, 120, 150, and 180 minutes (just before administration of [F]AV-133 radiopharmaceutical), 90, 120, 150, and 180 minutes (end of scan). PK samples were approximately 1 mL each (8 mL total including pre-dose sample). PK sample collection times were based on the start of NBI-750142 bolus administration.

[0173] NBI-750142 concentrations in cynomolgus monkey plasma samples were analyzed using a qualified LC-MS / MS bioanalytical method. The quantification range of the assay is 0.25-250 ng / mL, with dilution integrity demonstrated up to 1250 ng / mL. Plasma samples were processed using protein precipitation extraction with 0.1% formic acid in acetonitrile and addition of stable deuterium isotope-labeled internal standard NBI-750142-D6. Processed samples were chromatographically separated on a C18 column and interfaced to a Sciex API 4000 triple quadrupole mass spectrometer. Concentrations of NBI-750142 are shown in Tables 2-6.

[0174] Determination of NBI-750142 in cynomolgus monkey K2-EDTA plasma by LC-MS-MS The method used in this study was qualified in the range of 0.250–250 ng / mL based on the analysis of 100 μL of plasma by LC-MS-MS. Quantification was performed using a weighted 1 / x2 linear least-squares regression analysis generated from calibration standards. The protein precipitation extraction procedure was initiated by the addition of an internal standard solution (20.0 μL of deuterated NBI-750142-D6 at approximately 100 ng / mL). After the addition of 400 μL of acetonitrile / formic acid (1000:1.00), the tubes were sealed, vortexed, and centrifuged. Then, 100 μL of the supernatant was transferred to a new plate containing 400 μL of water / formic acid (1000:1.00). Chromatographic separation was achieved using an HPLC Kinetex C18 column (2.6 μm, 2.1 × 50 mm) with gradient elution using mobile phase A (water / formic acid; 1000:1.00) and mobile phase B (acetonitrile / methanol / formic acid; 500:500:1.00). A triple quadrupole mass spectrometer (Sciex API 4000) equipped with a TurboIonSpray source was used in positive ion mode. Quantitation was based on multiple reaction monitoring (MRM) of the m / z transitions of NBI-750142 and NBI-750142-D6.

[0175] Visualization of tissue uptake A volume of interest (VOI) defined within the cynomolgus monkey template space (see, for example, Ballanger et al. Neuroimage, 2013, 7:26-43) was applied to the spatially normalized PET images to define the caudate nucleus (0.96 cm 3 ), putamen (1.34cm 3 ) and cerebellum (3.81 cm 3 ) for time-activity curve (TAC, kBq / cm 3 ) was calculated. TAC and images (averaged over 60–120 min) are presented in standard uptake value (SUV) units (g / mL) normalized by animal weight and injected dose.

[0176] Image quantification Non-invasive Logan graph analysis (start time of fit, t *= 35 min) was used to estimate regional non-displaceable binding potential (BPND) using the cerebellar cortex as the reference TAC and the caudate and putamen TACs. BPND was calculated in the caudate and putamen across baseline and NBI-750142 scans. Target occupancy was calculated according to Equation 1-1.

number

number

[0177] result Three cynomolgus monkeys were randomly assigned to receive NBI-751042 at baseline and after administration of NBI-751042. 18 The relationship between the mean total NBI-750142 concentration during the scan (mean concentration from 60 to 180 min after administration of NBI-750142) and VMAT2 occupancy in the striatum (averaged occupancy from the caudate and putamen) was examined using [F]AV-133 (3.33 ± 0.45 mCi). Table 2-2 shows the relationship between the mean total NBI-750142 concentration during the scan (mean concentration from 60 to 180 min after administration of NBI-750142) and VMAT2 occupancy in the striatum (averaged occupancy from the caudate and putamen) for each evaluation. 18 F]AV-133 and NBI-750142 injection information is shown. Throughout the evaluation, 18 F]AV-133 SUV images are shown in Figures 1A-1D, and TAC is shown in Figure 2. At baseline, [F]AV-133 SUV images in the caudate and putamen were 18 Uptake of [F]AV-133 was higher in the sham-treated rats than in the rest of the brain. 18A dose-dependent decrease in [F]AV-133 binding was observed following administration of NBI-750142. BPND and occupancy are shown in Tables 2-3, 2-4, and 2-5 for assessment in animals LC206, EC865, and GC786, respectively. Total NBI-750142 concentrations and [ 18 The striatal occupancy of [F]AV-133 is shown in Tables 2-6. The relationship between occupancy of NBI-750142 and total plasma concentration is shown in Figure 3A, where the 50% effective total concentration (EC 50 ) was 11ng / mL. [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0178] conclusion The concentration-dependent occupancy of NBI-750142 at VMAT2 in NHP brain was 18 F]AV-133 and PET. Hill slope of 1 E max Using the model, occupancy was associated with total plasma NBI-750142 concentrations and EC 50was 11 ng / mL. Following the same process used for NBI-98782 (see Example 1), the PET method described above was applied to the human pharmacokinetic data to estimate VMAT2 target occupancy (%TO) for NBI-750142 (see FIG. 3B). It was determined that 60 mg BID of NBI-750142 (a dose predicted to be the maximum tolerated dose in humans) is predicted to achieve a lower %TO than the moderately effective dose of valbenazine. Thus, NBI-750142, at its maximum tolerated dose, would not be expected to be as effective as valbenazine in treating TD. These data serve as an example in which the process described herein can be used to evaluate VMAT2 compounds for further development at an early stage, enabling effective and efficient decision-making for the clinical development of VMAT2 inhibitors. Without being bound by theory, it is believed that the process described in the Examples (e.g., matching cynomolgus monkey PET with human PK data) may be useful at least for developing tetrabenazine analog VMAT2 inhibitors.

[0179] Example 3 [ 18 An Open-Label Positron Emission Tomography Study in Healthy Adult Male Subjects to Investigate VMAT2 Target Occupancy of a Single Oral Dose of NBI-750142 Using [F]AV-133 This study was conducted to evaluate VMAT2 brain penetration and target engagement after a single oral dose of NBI-750142 in healthy subjects. In this PET study, NBI-750142 engagement with VMAT2 in vivo was evaluated at two time points after dosing. The sequential cohort design allowed for the evaluation of safety, target occupancy and available pharmacokinetic (PK) data from each previous cohort before the selection of dose levels for the next cohort. A range of NBI-750142 doses (up to 200 mg) were tested to investigate the relationship between NBI-750142 plasma concentrations and VMAT2 engagement in selected brain regions.

[0180] This test is 18This was a single-center, open-label, single-dose, target occupancy Phase I study of NBI-750142 in up to 12 healthy male subjects using [F]AV-133 PET imaging. The study had a sequential cohort design with up to six cohorts, with 2-4 subjects per dose cohort receiving a single oral dose of NBI-750142. A total of 12 subjects completed the study. Subjects enrolled in the study were required to report to the clinical trial site in four waves, as detailed below.

[0181] Screening period (day -28 to day -2) The dosing day was day 1. Day -1 was the day before dosing, Day -2 was 48 hours before dosing, and Day -n was n days before dosing. Screening assessments were performed to determine eligibility (within 28 days prior to dosing of study drug). All screening safety assessments were performed to ensure subjects were medically healthy. Brain magnetic resonance imaging (MRI) was obtained in all subjects to assess eligibility. MRI was also utilized for anatomical localization and ROI (region of interest) analysis by co-registration to the subject's PET summation images.

[0182] If a subject was deemed eligible but did not enroll within the 28-day screening period, rescreening during the study was permitted. During the rescreening visit, subjects had all screening safety assessments repeated per protocol to ensure they were medically well, with no new clinically significant history or physical findings, laboratory profile, vital signs, or clinically significant findings on ECG. Brain MRI was not part of the safety assessment panel, so there was no need to repeat the MRI if it was performed within 1 year of the rescreening visit.

[0183] After completing the screening assessment, eligible subjects were instructed to: Refrain from taking prohibited drugs -Avoid alcohol and caffeinated products for 48 hours prior to Day 1 (measured from the day of dosing) Return to the study center for baseline evaluation between days -14 and -1 (measured from the day of dosing)

[0184] Baseline assessment (day -14 to day -1) Subjects underwent baseline assessment procedures within 14 days prior to Day 1 (day of dosing). On the day prior to the baseline visit, subjects were admitted to the clinical trial center. Subjects were medically evaluated and confirmed to meet study inclusion / exclusion criteria. Upon admission, a urine drug screen (UDS), urine cotinine, and urine alcohol test were performed. The following day, baseline [ 18 F]AV-133 PET scan was performed. If NBI-750142 administration occurred on the day after the baseline visit, subjects were not discharged between the baseline assessment and NBI-750142 administration.

[0185] Day before administration (Day -1) Subjects were admitted to the study center the day before NBI-750142 administration (day −1) if they were not already there and were discharged on day 3.

[0186] Treatment and follow-up period (Days 1-3) Two subjects were initially enrolled in each dose cohort. Each subject underwent one structural brain magnetic resonance imaging (MRI) scan during screening and one 18 Subjects underwent up to three [F]AV-133 PET scans (one baseline scan and up to two follow-up scans after receiving a single dose of NBI-750142). Subjects enrolled in the first cohort received a single dose of 100 mg of NBI-750142 orally on day 1. PET scans were performed approximately 1.5 hours after NBI-750142 administration on day 1 and approximately 18 hours after NBI-750142 administration on day 2. Subjects were subsequently discharged from the clinical trial center after all procedures and evaluations were completed on day 3.

[0187] Final study visit (day 10 ± 2 days or early termination) Subjects returned to the site for a final study visit on Day 10 (± 2 days). Safety assessments were performed, including physical and neurological examinations, adverse event assessments, electrocardiograms, vital signs, and laboratory tests.

[0188] Inclusion and Exclusion Criteria Subjects were eligible for the study if they met the following inclusion criteria: 1. Male subjects aged 18-55 years at the time of screening. 2. Medically healthy with no clinically significant medical history or clinically significant findings on physical examination, laboratory profile, vital signs or electrocardiogram at screening or on day -1. 3. Body mass index (BMI) of 18 kg / m 2 More than 30kg / m 2 It is less than or equal to. 4. Male subjects and their fertile partners must commit to using two forms of contraception, one of which is a barrier method for the male subject for the duration of the study. 5. Male subjects must not donate sperm during the study or for 90 days after completion of the study. 6. Be able to read, understand, sign and date the Informed Consent Form (ICF). 7. Willing to comply with all study procedures and restrictions, including refraining from strenuous, unaccustomed exercise and sports. 8. Agree to remain solely on the investigational unit for the required period and to return for all follow-up visits.

[0189] To be eligible for the study, subjects were required to not meet any of the following exclusion criteria: 1.Has had an unstable psychological disorder within the year prior to screening or is at significant risk for suicidal or violent behavior. 2. History of seizures, epilepsy, significant brain injury or lesion, stroke or transient ischemic attack, or intracranial or brain surgery. 3. Current clinically significant cardiovascular disease or abnormal screening ECG, including but not limited to a mean QT interval corrected three times using the Fridericia formula (QTcF) >450 msec at screening or baseline or a history of long QT syndrome. 4. A positive human immunodeficiency virus antibody (HIV-Ab) test result, a positive hepatitis B surface antigen (HBsAg) test result, or a positive hepatitis C virus antibody (HCV-Ab) test result and a positive HCV-Ab polymerase chain reaction (PCR) result at screening, or a history of positive results. 5. Hemoglobin level <12 g / dL at screening. 6. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT), or total bilirubin levels above the upper limit of normal (ULN) at screening. Subjects with a documented diagnosis of Gilbert syndrome do not need to meet the bilirubin criteria. 7. Have a positive alcohol urine test or UDS (positive for amphetamines, barbiturates, benzodiazepines, phencyclidine, cocaine, opiates, or cannabinoids) at screening or on day -1. 8. Consuming more than 2 alcoholic drinks per day or more than 14 alcoholic drinks per week within the 7 days preceding Day -1, or consuming any alcohol within the 48 hours preceding Day -1. 9.- Have used nicotine or cannabis products within 60 days prior to Day 1. 10. Use of any prescription or over-the-counter (OTC) medications within 7 days prior to Day -1, except for acetaminophen (within recommended doses). 11. Used alternative / complementary medicine products (e.g., herbal supplements, medicinal teas, creatine, sports supplements) within 7 days prior to Day -1, except for vitamins and minerals (supplemented with herbal preparations). Vitamins and minerals must be within the daily recommended dietary allowance (RDA) dose (e.g., daily multivitamins). 12. Use of strong inducers of CYP3A4 / 5 (e.g., rifampin, carbamazepine, phenytoin, phenobarbital, rifabutin, primidone, St. John's wort) within 30 days prior to day -1. 13. Have taken a strong CYP3A4 / 5 inhibitor (e.g., ketoconazole, itraconazole, erythromycin, clarithromycin, ritonavir) within 14 days or 5 half-lives (whichever is longer) prior to day -1. 14. Have taken a CYP2D6 inhibitor (e.g., bupropion, fluoxetine, paroxetine, quinidine) within 14 days or 5 half-lives (whichever is longer) prior to day -1. 15. Have taken a monoamine oxidase inhibitor (MAOI) (e.g., isocarboxazid, tranylcypromide, phenelzine, selegiline, rasagiline) within 30 days prior to Day -1. 16. Have taken a VMAT2 inhibitor (e.g., valbenazine, tetrabenazine, deutetrabenazine, or reserpine) within 30 days prior to Day -1; at any time [ 18 F]AV-133 injection or NBI-750142. 17. Baseline [ 18 Within 30 days prior to F]AV-133 injection, 18 Taking any of the medications listed under Concomitant Treatments that may interfere with [F]AV-133 PET imaging: methylphenidate, reserpine, amphetamine derivatives, dextroamphetamine derivatives, methylphenidate derivatives, or bupropion. 18.Currently taking medications known to cause QT prolongation. 19. Any other medical or psychiatric condition or laboratory abnormality that the investigator determines may prevent participation in the trial. 20. History of allergic reaction to VMAT2 inhibitors (e.g., valbenazine, tetrabenazine, deutetrabenazine, or reserpine). 21. Current substance abuse (e.g., painkillers, tranquilizers, opioids, stimulants, mood regulators) or known drug dependence. 22. Self-reported consumption of >5 caffeinated beverages per day in the past 30 days prior to Day -1 or self-reported consumption of caffeinated products within the 48 hours prior to Day -1. 23.- Consumed grapefruit juice or grapefruit products within 7 days prior to Day 1. 24.Has received any investigational product within 5 half-lives of the product if known, or at least 60 days prior to Day -1. 25. History or suspected poor compliance in clinical trials. 26.- Have blood loss or blood donation of 500 mL or more within 56 days prior to Day 1. 27. Brain MRI shows any of the following: infection, space-occupying lesion, normal pressure hydrocephalus, or evidence of any other abnormality related to CNS disease. 28. Have an implant, e.g., an implantable cardiac pacemaker or defibrillator, insulin pump, cochlear implant, metallic intraocular foreign body, implantable neurostimulator, CNS aneurysm clip, or other medical implant not certified for MRI, or have a history of claustrophobia with MRI. 29. In addition to the radiation exposure anticipated from your participation in this clinical trial, you have participated in other research protocols or clinical treatments within the past year that may have resulted in radiation exposure exceeding the annual permissible limit of 50 mSv effective dose set by U.S. federal guidelines. 30. The investigator has determined that the patient is not eligible to participate in the clinical trial.

[0190] Combination treatment All prescription medications, OTC medications, dietary supplements (including vitamins) and herbal supplements taken by subjects in the 30 days prior to screening were recorded. Use of prescription medications, OTC medications and alternative / complementary medicine products (e.g., herbal supplements, medicinal teas, creatine, sports supplements) was prohibited from within 7 days prior to day -1 until the end of the study, except for vitamins and minerals (unless supplemented with herbal preparations) and acetaminophen (within recommended doses). Vitamins and minerals must be within the daily RDA (recommended dietary allowance) doses (e.g., daily multivitamins).

[0191] The following drugs were prohibited in this study: · Strong inducers of CYP3A4 / 5 (e.g., rifampin, carbamazepine, phenytoin, phenobarbital, rifabutin, primidone, St. John's wort) within 30 days prior to Day 1. · Strong CYP3A4 / 5 inhibitors (e.g., ketoconazole, itraconazole, erythromycin, clarithromycin, ritonavir) within 14 days or 5 half-lives (whichever is longer) prior to day -1. · CYP2D6 inhibitors (e.g., bupropion, fluoxetine, paroxetine, quinidine) within 14 days or 5 half-lives (whichever is longer) prior to Day -1. -MAOIs (e.g., isocarboxazid, tranylcypromide, phenelzine, selegiline, rasagiline) within 30 days of Day 1. - Within 30 days prior to day -1, VMAT2 inhibitors (e.g., valbenazine, tetrabenazine, deutetrabenazine, or reserpine) and [18F]AV-133 injection or NBI-750142 at any time point. -Prescription or OTC medications, except acetaminophen (within recommended doses), within 7 days prior to Day 1. - Alternative or complementary medical products (e.g. herbal supplements, medicinal teas, creatine and sports supplements), other than vitamins and minerals (except those supplemented with herbal preparations) within the daily recommended daily allowance, within 7 days prior to Day 1.

[0192] The following list of medicines can be found at 18 F]AV-133 may interfere with PET imaging and may increase baseline [ 18 Within 30 days prior to [F]AV-133 injection, the following were not permitted: Methylphenidate VMAT2 inhibitors Amphetamine derivatives Dextroamphetamine derivatives Methylphenidate derivatives Bupropion

[0193] Dietary and other restrictions Subjects were fasted for the duration of the study. Subjects were required to abstain from large meals for at least 8 hours prior to NBI-750142 administration. If fasting was greater than 8 hours, subjects consumed a full breakfast (given 8 hours prior to dosing). All subjects consumed a small, prescribed snack at least 2 hours prior to NBI-750142 administration. Subjects were not permitted to consume any food for at least 3 hours after NBI-750142 administration and were not permitted to consume food during the PET scan.

[0194] Subjects were not permitted to consume water (except for the 240 mL required for administration) from 1 hour prior to until 1 hour after NBI-750142 administration. Water was permitted during all other fasting periods. Subjects were also required to fast before the baseline and second post-dose PET scans as well as before the first post-dose PET scan.

[0195] Other dietary restrictions included: ·alcohol Caffeine products Nicotine or cannabis products ·Grapefruit or grapefruit juice

[0196] NBI-750142 study drug NBI-750142 was made into a solution consisting of water within 72 hours of dosing. The appropriate volume of dosing solution (based on cohort) was administered via an oral dosing syringe. After dosing, subjects were instructed to ingest the prescribed volume of water to bring the total dosing volume of solution and water to 240 mL. Subjects were required to ingest the dosing solution and water within 2 minutes.

[0197] [ 18 [F]AV-133 Imaging Procedure The subject was positioned in a gently fixed head holder using the camera's laser light so that the brain was in the center of the field of view. Prior to injection of the radiotracer and emission imaging, a CT scan was performed to obtain a correction factor for photon attenuation by the material.

[0198] Subjects were administered 100 mg of 10 ... 18 A single dose of [F]AV-133 was administered intravenously over 3 min, followed by a 10 mL saline flush. Dynamic PET images of the brain were acquired over 120 min following radiotracer injection into a Siemens Biograph 6 PET / CT camera according to the following protocol: Head CT scan ·[ 18 F]AV-133 injection (T=0 minutes) Emission scan: 6 x 30 seconds, 4 x 1 minute, 4 x 2 minutes, 21 x 5 minutes (0-120 minutes)

[0199] Images were reconstructed into a 168x168x81 matrix (pixel size 2.03mmx2.03mm) using an iterative reconstruction algorithm (OSEM 4 iterations, 16 subsets) and a posterior Gaussian filter = 3mm. Standard corrections for random, scattering, system dead time and attenuation provided by the camera manufacturer were performed.

[0200] MRI brain imaging MRI scans were obtained from eligible subjects as part of the screening visit to identify and delineate brain anatomical regions of interest for individual PET images. MRI scans were obtained on a Siemens Espree 1.5 Tesla clinical magnet. MRI scans of 1 mm contiguous slices were obtained in the following order: sagittal T1, horizontal T2 double echo, horizontal flair, coronal T2, horizontal DWI, horizontal T1, sagittal MPRAGE.

[0201] [ 18 F]AV-133 image analysis Reconstructed PET image data volumes were transferred to the image processing PMOD software package (PMOD Technologies, Zurich, Switzerland), where images were motion and attenuation corrected, reregistered onto the subject's MRI, and subsequently normalized to standard MNI (Montreal Neurological Institute) space, where volumes of interest (VOIs) were defined from a template (see, for example, Hammers et al, Adult brain maximum probability map ("Hammersmith atlas"; n30r83) in MNI space, 2008). Subject MRIs were segmented into gray matter, white matter, and CSF maps. The mean activity concentration (kBq / cc) within each VOI, confined to gray matter voxels in cortical regions, was measured and time-activity curves (TACs) were generated. TACs were extracted from the following VOIs: caudate and putamen (striatum), midbrain, cerebellum, and occipital cortex. TACs (TACs and ) were expressed in standard uptake value (SUV) units (g / mL) by normalizing to subject weight and injection dose. The activity of the target regions was normalized to generate SUVr images, using the occipital cortex as the reference region. For visualization, standard uptake value ratio (SUVr) images were calculated between 60 and 120 min. The primary outcome measure, non-displaceable binding potential (BPND), was evaluated by non-invasive Logan graph analysis (NI-LGA, t *= 20 min). The measured occupancy percentage (OM) of NBI-750142 (drug) by region was calculated according to Equation 3-1 at each post-administration time point, T1 (1.5 hours post-administration) and T2 (18 hours post-administration).

number

[0202] E in Equation 3-2 max A model was used to describe the relationship between plasma concentrations of NBI-750142 (ng / mL, average of start-scan and end-scan measurements) and occupancy in the striatum (average occupancy of the caudate and putamen) and midbrain, where E max is limited to 100, i.e., at a sufficiently high concentration the maximum occupancy reaches 100%,

number

[0203] In post-hoc exploratory analyses, an upregulation model (Equation 3-3) was applied to explain the apparent "negative occupancy" values ​​observed at T2 (BPND values ​​at T2 were unexpectedly higher than the corresponding BPND values ​​at baseline).

number

[0204] Test results Target Demographics A total of 20 healthy male volunteers were screened and a total of 12 participants were enrolled. Participants' ages at screening ranged from 29 to 54 years (mean 43.2 years). Nine of the enrolled subjects were black or African American and three were white. One was Hispanic and 11 were non-Hispanic. Subject demographics are shown in Table 3-1. [Table 3-1]

[0205] [ 18 [F]AV-133 brain imaging and time-activity curves Twelve healthy male volunteers were randomly assigned to 18 Subjects were imaged with [F]AV-133 at baseline and at two time points after NBI-750142 administration, 1.5 hours (T1) and 18 hours (T2). Each cohort had 2-4 subjects: Cohort 1 received 100 mg; Cohort 2 received 200 mg; Cohort 3 received 60 mg; Cohort 4 received 60 mg; Cohort 5A (subjects 1 and 2) received 10 mg; Cohort 5B (subjects 3 and 4) received 10 mg.

[0206] The 60 mg cohorts had different dilutions (cohort 3 = 20 mg / mL in 3 mL, cohort 4 = 10 mg / mL in 6 mL). For each cohort, the mean dilution time was averaged between 60 and 120 minutes. 18 [F]AV-133 SUVr images (normalized to the occipital cortex) are shown in Figures 7A-7F for baseline and post-NBI-750142 administration (T1 and T2). 18 F]AV-133 SUVr TAC is shown in Figures 8A-8F for the same imaging time points for each cohort. 18F]AV-133 uptake was higher in the caudate nucleus and putamen (target regions) where VMAT2 is abundant, compared to the rest of the brain. TAC showed relatively low activity in the midbrain and cerebellum, with the lowest uptake in the occipital cortex. At T1, [F]AV-133 uptake in the striatum of each subject was 18 Compared to baseline [F]AV-133, the 100 mg and 200 mg doses significantly reduced uptake in both subjects; 60 mg in cohorts 3 and 4 showed variability between subjects with a smaller reduction in striatal uptake in subject 2 compared to subject 1 in both cohorts; 10 mg in cohorts 5A and 5B showed no reduction in uptake. Across all cohorts, striatal uptake at T2 appeared to return to or slightly exceed baseline levels.

[0207] Quantitative evaluation The primary outcome measure, BPND, was estimated using NI-LGA, with the occipital cortex as the reference region. BPND was estimated in the target regions, the caudate and putamen, and in the less connected region, the midbrain. The cerebellum was [ 18 Cerebellar BPND was not evaluated because it is also a known reference region for [F]AV-133. In addition, slightly higher uptake in the cerebellar cortex compared to the occipital cortex was observed in SUVr images and TAC. 18 The BPND of [F]AV-133 and VMAT2 occupancy of NBI-750142 at T1 and T2 are shown in Table 3-2. The occupancy at T2 in the striatum of subject 1 and subject 2, respectively, was as follows: 100 mg cohort 1: 1% and 14%; 200 mg cohort 2: -28% and -6%; 60 mg cohort 3: -14% and -21%; 60 mg cohort 4: -10% and -12%; 10 mg cohort 5A: -3% and -4%; and 10 mg cohort 5B: -14% (subject 3) and 5% (subject 4).

[0208] The occupancy in the midbrain reflects the striatum, where (positive) occupancy was observed in T1; however, an apparent negative occupancy was calculated in T2. max A model (see Equation 3-2) was used to describe the relationship between plasma concentrations of NBI-750142 and striatal occupancy of VMAT2, resulting in EC 50 9A-9C show the plasma concentrations of NBI-750142 plotted against T1 and T2 occupancy, T1 occupancy only, and T2 occupancy only. EC 50 was estimated to be 25.5 ng / mL when only T1 occupancy values ​​were used and 35.2 ng / mL when both T1 and T2 occupancy values ​​were used. max The model could not fit the occupancy data at T2 alone. [Table 3-2]

[0209] Post-hoc analysis: Up-regulation model Apparent negative occupancy was observed primarily at T2 among the cohorts, with E max Given our inability to fit the model to the T2 data, we assessed post hoc exploratory analyses applying the upregulation model (Eq. 3-3; see also Gunn and Rabiner, Semin. Nucl. Med. 2017, 47: 89-98). The negative occupancy at T2 was attributed to higher striatal BPND at T2 compared to baseline. max A comparison of data fits using the model (Eq. 3-2) and the enhanced upregulation model (Eq. 3-3) is shown in Figure 10A. The upregulation model uses both T1 and T2 data to estimate the EC 50 The upregulation model estimated the α-to-α ratio (α = 6.3 ng / mL) and the URF of 2.76. The upregulation model was used to fit the data using both T1 and T2 occupancy as a function of plasma concentration of NBI-750142 with an E(2) score based on visual assessment of fit and the corrected Akaike's Information Criterion (AICc). maxCompared with the model, it better described the upregulation model (AICc = 123 vs E max Model: AICc=131). The AICc was calculated for each fit to the T1 data only. max However, as shown in Figure 10B, the upregulation model showed a lower EC 50 is estimated.

[0210] Summary and Conclusion In this Phase 1, open-label study, 12 healthy adult male subjects were 18 Vesicular monoamine transporter 2 (VMAT2) target occupancy was examined after a single oral dose of NBI-750142, as imaged by [F]AV-133. Occupancy in the striatum (mean occupancy in caudate and putamen) ranged from -4% (10 mg) to 76% (100 mg) at T1 and from -28% (200 mg) to 14% (100 mg) at T2. The relationship between VMAT2 target occupancy and NBI-750142 plasma concentrations was examined using E max The model was evaluated. EC 50 was estimated to be 25.5 ng / mL using T1 occupancy and 35.2 ng / mL using both T1 and T2 occupancy. However, EC 50 could not be estimated from T2 occupancy data alone.

[0211] Although plasma concentrations of NBI-750142 were above the level of quantification at T2, ranging from approximately 0.3 to 10 ng / mL, many calculated T2 occupancies were negative. Negative occupancies observed in PET studies can arise when baseline levels of binding (BPND) at the time of the "drug" scan no longer match previous baseline scan measurements. 18The test-retest value for BPND of [F]AV-133 is 9.4% (see, e.g., Freeby et al., Mol. Imaging Biol. 2016, Apr;18(2):292-301). This suggests that the trend for negative occupancy at T2 observed in this study is different from 0%. Without being bound by theory, one possible explanation for this discrepancy is an upregulation of the number of VMAT2 binding sites accessible to the radiotracer following drug administration. To address this hypothesis, a post hoc exploratory analysis using the upregulation model was performed. Analyses using both T1 and T2 occupancy showed that EC 50 was estimated to be 6.3 ng / mL, and the upregulation factor was 2.76. max Compared with the model, the upregulation model better described the relationship between NBI-750142 plasma concentrations and T1 and T2 occupancy. This conclusion was based on visual assessment of model fit and the "corrected AIC" standard goodness-of-fit metric (upregulation model: AICc = 123 vs E max Model: AICc = 131). AIC is a measure of goodness of fit that is penalized for the number of parameters. Corrected AIC is appropriate for small "n" studies. This result is consistent with E max The model is consistent with the true EC 50 Larger than EC 50 This suggests that it may provide value.

[0212] In conclusion, substantial VMAT2 occupancy by NBI-750142 was observed in [ 18 F]AV-133 and PET imaging were used in living human brains from healthy volunteers. max Applying the model to only the T1 data (1.5 hours after NBI-750142) gave an estimated EC 50 was obtained, but E maxExploratory analyses of all data (from T1 and T2) were also performed using a model that allowed for upregulation of VMAT2 sites after drug administration. EC from the latter analysis 50 Although exploratory, the findings of this upregulation model are supported by E from T1 data alone. max This suggests that the results should be treated as an upper limit.

[0213] As outlined in Example 2 (see, e.g., FIG. 3A), the EC of NBI-750142 in cynomolgus monkey PET 50 The EC value was 11 ng / mL, which is lower than that obtained in humans using two different models. 50 Because NBI-750142 PPB in cynomolgus monkeys and humans is similar, this close relationship between monkey and human PET results is consistent with the EC 50 This also applies to values ​​for PET-derived EC in humans and cynomolgus monkeys. 50 The fairly good agreement between the PET EC 50 The values ​​were applied to human PK data, further supporting the conclusions described in Examples 1-2.

[0214] Example 4 [ 11 Effects of valbenazine on the dopamine system in humans measured by [C]-PHNO positron emission tomography (PET) Interim imaging and tolerability data from the ongoing study were collected and analyzed in a cohort of two to four healthy volunteers. For each scan, participants received a radioligand that is a D2 / D3 dopamine receptor agonist [ 11 C](+) 4-Propyl-3,4,4a,5,6,10b-hexahydro-2H-naphtho[1,2-b][1,4]oxazin-9-ol ([ 11Participants received an injection of [C]-PHNO followed by 90 minutes of data acquisition using a Siemens Biograph PET / CT. For the post-valbenazine scan, participants 11 Valbenazine was administered orally 6–8 h before [C]-PHNO administration, and PET imaging was performed around the time when plasma concentrations of valbenazine were maximal. ND ) was used as the primary endpoint. Using the cerebellum as the reference region, regional BP ND The decrease in synaptic dopamine after valbenazine administration was [ 11 C]-PHNO BP ND Increase in (ΔBP ND ) corresponded to the mean plasma (+)-α-dTBZ concentration (C ave ) for each participant's ΔBP ND and obtained exposure-response curves.

[0215] Nine participants (five men and four women) were administered valbenazine at doses ranging from 40 to 160 mg, and the C ave The mean mean mean blood glucose concentration was approximately 10-60 ng / mL. In the interim analysis, 8 participants showed a dose-dependent 11 C]-PHNO ΔBP ND The higher the exposure to (+)-α-dTBZ with higher doses of valbenazine, the greater the increase in ΔBP ND The adverse events in this study were consistent with the known safety and tolerability profile of valbenazine, as previously reported in clinical trials for tardive dyskinesia (TD).

[0216] Valbenazine is a 11 C]-PHNO ΔBP NDThe approximately 20-40% [ 11 C]-PHNO ΔBP ND Increases in dopamine levels have previously been seen following treatment with tyrosine hydroxylase inhibitors, which deplete dopamine. 11 C]-PHNO ΔBP ND (Caravaggio et al, Neuropsychopharmacology 2014;39:2769). Thus, biologically meaningful dopamine reductions were observed in humans at pharmacological and therapeutic doses of valbenazine. These data enable future investigations into the relationship between VMAT2 inhibition and the potential treatment of other CNS disorders.

[0217] Other embodiments Although the present invention has been described with its detailed description, it should be understood that the foregoing description is intended to illustrate, but not to limit, the scope of the invention as defined by the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. It should be understood by those skilled in the art to which the present invention pertains that any of the features described herein with respect to any particular aspect and / or embodiment of the present invention may be combined with any one or more of any other features of any other aspect and / or embodiment of the present invention described herein, with appropriate modifications to ensure compatibility of the combination. Such combinations are part of the invention contemplated by this disclosure.

Claims

1. 1. A method for preparing a pharmaceutical composition comprising a therapeutically effective dose of a VMAT2 inhibitor, comprising: (1) measuring in vivo VMAT2 occupancy of the VMAT2 inhibitor in a subject to which an amount of the VMAT2 inhibitor has been administered, wherein a VMAT2 occupancy of 80-96% indicates that the amount of the VMAT2 inhibitor is a therapeutically effective dose; and (2) mixing the therapeutically effective dose of the VMAT2 inhibitor with a pharmaceutically acceptable carrier. A method comprising:

2. A method for identifying a therapeutically effective dose of a VMAT2 inhibitor, comprising: (1) measuring in vivo VMAT2 occupancy of the VMAT2 inhibitor in a subject to which the VMAT2 inhibitor has been administered; and (2) identifying a therapeutically effective dose of a VMAT2 inhibitor when the VMAT2 occupancy rate of the amount of the VMAT2 inhibitor is 80 to 96%; A method comprising:

3. A method for preparing a pharmaceutical composition comprising a therapeutically effective dose of a VMAT2 inhibitor, comprising: mixing said therapeutically effective dose of said VMAT2 inhibitor with a pharmaceutically acceptable carrier. Including, the therapeutically effective dose of the VMAT2 inhibitor is identified by measuring in vivo VMAT2 occupancy of the VMAT2 inhibitor in a subject previously administered an amount of the VMAT2 inhibitor; A VMAT2 occupancy of 80-96% indicates that said amount of said VMAT2 inhibitor is a therapeutically effective dose.

4. VMAT2 occupancy is measured by one or more imaging techniques; the one or more imaging techniques comprise administering to the subject an imaging agent capable of binding to VMAT2, and thereafter imaging the subject; The method of any one of claims 1 to 3, wherein the imaging agent is a VMAT2 inhibitor.

5. VMAT2 occupancy is measured by positron emission tomography (PET) assay; 5. The method of any one of claims 1 to 4, wherein the PET assay comprises administering to the subject a PET imaging agent capable of binding to VMAT2, and thereafter imaging the subject.

6. The PET assay comprises: (a) administering to said subject a PET imaging agent capable of binding to VMAT2; (b) waiting a sufficient time for the PET imaging agent to bind to VMAT2; (c) imaging the object one or more times; (d) measuring VMAT2 displacement of said PET imaging agent; and (e) determining VMAT2 occupancy based on the measured VMAT2 displacement of said PET imaging agent. Including, Optionally, said VMAT2 displacement of said PET imaging agent is measured at one or more time points during said imaging; 6. The method of claim 5, wherein the method further comprises imaging the subject prior to step (a) to obtain a baseline image. (i) the administration of the VMAT2 inhibitor to the subject occurs after step (a); (ii) said administering said VMAT2 inhibitor to said subject occurs after step (b); or (iii) the administering of the VMAT2 inhibitor to the subject occurs after step (b) and before step (c); The method of claim 6.

8. The method described in claim 6 or 7, further comprising a step of measuring the plasma concentration of the VMAT2 inhibitor in the subject. (i) the plasma concentration of the VMAT2 inhibitor is measured at one or more time points during the imaging of step (c). (ii) the plasma concentration of the VMAT2 inhibitor is measured at one or more time points prior to the imaging of step (c); (iii) the plasma concentration of the VMAT2 inhibitor is measured at one or more time points during and prior to the imaging of step (c); (iv) the plasma concentration of the VMAT2 inhibitor is measured at one or more time points from about 2 hours before the imaging of step (c) until the end of the imaging; or (v) the plasma concentration of the VMAT2 inhibitor is measured at one or more time points from about 1 hour before the imaging of step (c) to the end of the imaging; The method of claim 8.

10. the PET imaging agent is a radiolabeled VMAT2 inhibitor; as needed, the PET imaging agent is a [ 11 C]-radiolabelled VMAT2 inhibitor; or the PET imaging agent is a [ 18 F]-radiolabelled VMAT2 inhibitor; The method according to any one of claims 5 to 9.

11. the PET imaging agent is a radiolabeled analog of a VMAT2 inhibitor selected from the group consisting of valbenazine, tetrabenazine, deutetrabenazine, dihydrotetrabenazine, NBI-750142, and AV-133; or the PET imaging agent is a [ 11 C]- or [ 18 F]-radiolabelled analog of a VMAT2 inhibitor selected from the group consisting of valbenazine, tetrabenazine, deutetrabenazine, dihydrotetrabenazine, NBI-750142, and AV-133; Optionally, the radiolabeled analogue of dihydrotetrabenazine is a radiolabeled analogue of (+)-α-dihydrotetrabenazine. The method according to any one of claims 5 to 10.

12. The PET imaging agent is １８ 12. The method according to any one of claims 5 to 11, wherein the antibody is [F]-AV-133.

13. The method further comprising monitoring the subject for one or more symptoms associated with a treatment-emergent adverse event (TEAE) following administration of the VMAT2 inhibitor; as needed, the monitoring step occurs from about 30 minutes to about 90 minutes after administration of the VMAT2 inhibitor; or the monitoring step occurs from about 30 minutes to about 60 minutes after administration of the VMAT2 inhibitor. The method according to any one of claims 1 to 12.

14. The method further comprising identifying the subject as not exhibiting one or more symptoms associated with TEAEs after administration of the VMAT2 inhibitor; the one or more symptoms are selected from ptosis, hypoactivity, sedation, anxiety, nausea, akathisia, and salivation; The method according to any one of claims 1 to 13. (i) the administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 80% and not more than 95%; (ii) the administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 85% and not more than 95%; or (iii) the administered dose is identified as a therapeutically effective dose if VMAT2 occupancy is determined to be at least 85% and not more than 90%; The method according to any one of claims 1 to 14.

16. A method according to any one of claims 1 to 15, wherein the subject is a mammal.

17. A method according to any one of claims 1 to 15, wherein the subject is a human.

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