Pyridazinone compositions for the treatment of neuromuscular conditions

Abstract

Described herein is the treatment of neuromuscular diseases, such as Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), limb-girdle muscular dystrophy (LGMD), and McArdle's disease, with compositions comprising substituted pyridazinone compounds. Described herein is a method for treating neuromuscular conditions, comprising administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day. In some embodiments, the method comprises administering to a subject a compound of formula (I) at a dose of 15 mg to 40 mg per day.

Description

[Technical Field]

[0001] cross reference This application claims the benefit of U.S. Provisional Application No. 63 / 375,180, filed September 9, 2022; U.S. Provisional Application No. 63 / 375,366, filed September 12, 2022; U.S. Provisional Application No. 63 / 510,348, filed June 26, 2023; and U.S. Provisional Application No. 63 / 580,334, filed September 1, 2023, the entireties of which are hereby incorporated by reference herein. [Background technology]

[0002] Background of the Invention Skeletal muscle is the largest organ system in the human body and serves two primary purposes. The first is force generation, which enables muscle contraction, locomotor activity, and postural maintenance. The second is glucose, fatty acid, and amino acid metabolism. Skeletal muscle contraction during daily activities and exercise is naturally associated with muscle tone, breakdown, and remodeling, which are important for muscle adaptation. In individuals with neuromuscular conditions, such as Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), limb-girdle muscular dystrophy (LGMD), and McArdle disease, muscle contraction results in a continuous cycle of amplified muscle breakdown, which the body attempts to repair. Ultimately, as patients age, a pathophysiological process emerges that leads to excessive inflammation, fibrosis, and the accumulation of fat deposits in muscle, which are precursors to a rapid decline in physical function and a contribution to mortality. One potential treatment for such neuromuscular conditions is the administration of compounds that reduce skeletal muscle contraction. However, reduced skeletal muscle contractility can also lead to muscle weakness, which in turn can reduce a patient's physical performance. There remains a need for treatments that reduce muscle breakdown in patients with neuromuscular conditions while preserving or improving the patient's muscle strength. Summary of the Invention [Means for solving the problem]

[0003] Summary of the Invention 1. A method of treating a neuromuscular condition, comprising administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day: [ka]

[0013] Methods are described herein, comprising administering to a subject a compound of Formula (I) at a dose of 15 mg to 40 mg per day. In some embodiments, the methods involve administering to a subject a compound of Formula (I) at a dose of 15 mg, 20 mg, 25 mg, or 30 mg per day. In some embodiments, the methods involve administering to a subject a compound of Formula (I) at a dose of 20 mg per day. In some embodiments, the methods involve administering to a subject a compound of Formula (I) at a dose of 1 mg to 20 mg per day. In some embodiments, the methods involve administering to a subject a compound of Formula (I) at a dose of 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, or 20 mg per day. In some embodiments, the methods involve administering to a subject a compound of Formula (I) at a dose of 1 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 2 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 2.5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 7.5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 10 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 15 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 20 mg per day. In some embodiments, the subject is over 18 years of age. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 10 mg per day. In some embodiments, the method comprises administering to the subject a compound of formula (I) at a dose of 15 mg per day. In some embodiments, the method comprises administering to the subject a compound of formula (I) at a dose of 20 mg per day. In some embodiments, the subject is 18 years old or younger. In some embodiments, the method comprises administering to the subject a compound of formula (I) at a dose of 10 mg per day.In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 7.5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 2.5 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 2 mg per day. In some embodiments, the method comprises administering to the subject a compound of Formula (I) at a dose of 1 mg per day. In some embodiments, the method comprises administering a first dose of a fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose. In some embodiments, the method comprises administering, and maintaining or increasing the North Star Ambulatory Assessment (NSAA) score of the subject compared to the NSAA score before treatment. In some embodiments, the method maintains or improves one or more of the following functional measures in the subject: maximum elbow flexion strength compared to maximum elbow flexion strength before treatment; maximum knee extension strength compared to maximum knee extension strength before treatment; 10-meter walking / running speed compared to 10-meter walking / running speed before treatment; 100-meter timed test velocity compared to 100-meter timed test velocity before treatment; 4-step stair climbing timed velocity compared to 4-step stair climbing timed velocity before treatment; and maximum grip strength compared to maximum grip strength value before treatment. In some embodiments, the method maintains the creatine kinase activity level or reduces the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment. In some embodiments, the method maintains the creatine kinase activity level or increases the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment. In some embodiments, the method maintains the subject's dual-energy x-ray absorptiometry (DXA) lean mass % or increases the subject's DXA lean mass % by 5% or more compared to the DXA lean mass % before treatment.In some embodiments, the method maintains the subject's Fast Skeletal Muscle Troponin I (TNNI2) concentration or reduces the subject's TNNI2 concentration by 10% or more compared to the TNNI2 concentration before treatment, optionally the TNNI2 concentration is estimated from SOMAscan levels. In some embodiments, the administering reduces the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more compared to the protein concentration of one or more muscle injury biomarkers before treatment. In some embodiments, the one or more muscle injury biomarkers include ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof. In some embodiments, the administering reduces the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more pro-inflammatory proteins before treatment. In some embodiments, the one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In some embodiments, the administering increases the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more anti-inflammatory proteins before treatment. In some embodiments, the one or more anti-inflammatory proteins include IL10, IL13, IL22, IL37, IL4, or a combination thereof. In some embodiments, the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle disease. In some embodiments, the neuromuscular condition is Becker muscular dystrophy. In some embodiments, the neuromuscular condition is Duchenne muscular dystrophy.In some embodiments, administering comprises administering the compound of Formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more. In some embodiments, administering comprises administering the compound of Formula (I) for 4 months or more, 5 months or more, 6 months or more, or 7 months or more. In some embodiments, administering comprises administering the compound of Formula (I) for 6 months or more. In some embodiments, prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a)-(g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a dystrophin gene mutation of -0.1 to -10, or Further comprising selecting subjects for treatment who have one or more of the following: (a) pre-treatment NSAA annual change of -0.5 to -3; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher.

[0004] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or increase the subject's North Star Ambulatory Assessment (NSAA) score compared to the NSAA score before treatment.

[0005] Described herein are methods of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or improve one or more of the following functional measures in the subject: 100-meter timed test speed compared to 100-meter timed test speed before treatment; timed speed for climbing four steps compared to timed speed for climbing four steps before treatment; and maximum grip strength compared to maximum grip strength value before treatment.

[0006] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain creatine kinase activity levels or reduce the subject's creatine kinase activity levels by 10% or more compared to the creatine kinase activity levels before treatment.

[0007] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain serum creatinine levels or increase the subject's serum creatinine levels by 10% or more compared to the serum creatinine level before treatment.

[0008] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject an amount of a fast-twitch myosin (type II) inhibitor sufficient to maintain the subject's dual-energy x-ray absorptiometry (DXA) lean mass % or to increase the subject's DXA lean mass % by 5% or more compared to the DXA lean mass % before treatment.

[0009] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's fast skeletal troponin I (TNNI2) concentration or to reduce the subject's TNNI2 concentration by 10% or more compared to the TNNI2 concentration before treatment.

[0010] A method for inhibiting fast-twitch myosin (type II) in a subject in need thereof, comprising the steps of: (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; and (d) a treatment with a saturation of -0.1 to -10, -0.5 to -3, or -1 to -4. (e) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (f) DXA lean mass % of <75%, <70%, <65%, <60%; and (g) self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher; and administering to the subject a fast-twitch myosin (type II) inhibitor.

[0011] A compound of formula (I) in an amount of about 1 mg to about 20 mg [ka] Described herein are unit doses for oral administration comprising:

[0012] 1. A kit comprising an oral pharmaceutical composition and instructions for administration to a subject in need thereof, wherein the oral pharmaceutical composition comprises a compound of formula (I): [ka] and a pharmaceutically acceptable excipient.

[0013] 1. A method of treating a neuromuscular condition, comprising administering to a subject an AUC24 a compound of formula (I) in an amount sufficient to maintain [ka] The methods described herein include administering

[0014] Described herein are methods for treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain myoglobin levels or reduce the subject's myoglobin levels by 10% or more compared to the myoglobin levels before treatment.

[0015] Described herein are methods for reducing pain associated with a neuromuscular condition, comprising administering to a subject in need thereof a fast-twitch myosin (type II) inhibitor. Incorporation by Reference

[0016] All publications, patents, and patent applications mentioned herein are incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. Patent publications WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, WO2021231615, and PCT application PCT / US2023 / 021739 are incorporated by reference herein.

[0017] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings (also herein "Figure" and "FIG") [Brief explanation of the drawings]

[0018] [Figure 1] Figure 1 presents results from a study of BMD patients showing disease progression as measured by NSAA.

[0019] [Figure 2] FIG. 2 shows the NSAA scores of BMD patients after 4 months of Compound 1 administration.

[0020] [Figure 3] FIG. 3 depicts baseline characteristics for BMD patients compared with age-normal values.

[0021] [Figure 4] Figure 4 shows the plasma levels of Compound 1 and step counts in BMD patients after 4 months of dosing. The mean AUC for Compound 1 plasma PK levels was 2,293 ng / mL, 1,454 ng / mL, and 2,299 ng / mL for the 20 mg, 10 mg, and 15 mg doses, respectively.

[0022] [Figure 5] FIG. 5 depicts measurements of creatine kinase activity and fast skeletal troponin I (TNNI2) levels in BMD patients following administration of Compound 1 compared to baseline measurements.

[0023] [Figure 6] FIG. 6 depicts the NSAA scores of patients 2 and 4 months after the first dose of Compound 1 compared to baseline measurements.

[0024] [Figure 7] FIG. 7 depicts the increase in functional measures such as walking / running speed, stair climbing speed, and grip strength in BMD patients 2 and 4 months after the first dose of Compound 1 compared to baseline measurements.

[0025] [Figure 8] FIG. 8 depicts the reduction in self-reported pain scores in BMD patients 2 and 4 months after the first dose of Compound 1.

[0026] [Figure 9A] 9A, 9B, and 9C depict the North Star Ambulatory Performance Assessment. [Figure 9B] Same as above. [Figure 9C] Same as above.

[0027] [Figure 10AB] Figures 10A-10G show changes in several biomarkers after treatment with Compound 1. **p<0.01; ***p<0.001. Figure 10A shows the mean creatine kinase (CK) levels in different dosing regimens. All P<0.05 except for month 3. Figure 10B shows the mean fast skeletal troponin I (TNNI2) levels in different dosing regimens. All P<0.05 except for month 4. Figure 10C shows the mean myoglobin levels in different dosing regimens. All P<0.05 except for months 2, 3, 4, 6, or 10. Figure 10D shows the change in CK activity in each patient from the 12-month treatment group. p=0.0087. Figure 10E shows the mean CK levels between baseline and the 12-month treatment group. p=0.0012. The stated percentage differences were relative to the mean baseline. Bar graphs show the mean ± SEM. Figure 10F shows the change in TNNI2 activity for each patient in the 12-month treatment group. p=0.022. TNNI2 data were predicted from SOMAscan. Figure 10G shows the mean TNNI2 levels between baseline and the 12-month treatment group. p<0.0001. The percentage difference indicated was relative to the mean baseline. The graph shows the mean ± SEM. TNNI2 data were predicted from SOMAscan. [Figure 10C] Same as above. [Figure 10DE] Same as above. [Figure 10FG] Same as above.

[0028] [Figure 11AB]Figures 11A-11D show the change in NSAA score and the plasma concentration of Compound 1 under different dosing regimens, as well as their correlation. Figure 11A shows the change in NSAA score under different dosing regimens. The dashed line indicates the mean ± 95% CI. The downward trend callout reflects natural history data based on Luca Bello at al. MDA (2022) and van de Velde NM et al., Neurology (2021). Figure 11B shows the plasma concentration of Compound 1 under different dosing regimens. Figure 11C shows the NSAA response of individual patients from the 12-month group. The dashed line represents the expected natural history decline in untreated patients. Figure 11D shows the corresponding plasma concentration of Compound 1 from the 12-month group. [Figure 11CD] Same as above.

[0029] [Figure 12AB] Figures 12A-12F show functional assessments after treatment with Compound 1. Figure 12A shows the mean 10-meter speed in different dosing regimens. All p-values ​​are NS, with a 7% decrease at 12 months. Last observation carried forward (all N=12, except for two missing values ​​at 4 and 8 months). Figure 12B shows the mean 100-meter speed in different dosing regimens. All p-values ​​are NS, with a 0% change at 12 months. Last observation carried forward (all N=12, except for two missing values ​​at 4 and 8 months). Figure 12C shows the mean 4-step stair climbing time speed in different dosing regimens. All p-values ​​are NS, except for the p-value at 12 months, which was 0.016, with a 13% decrease at 12 months. Figure 12D shows the mean maximum grip strength in different dosing regimens. p-values ​​are <0.05 at 4, 6, 8 months, and NS at 12 months. There was an 8% decrease at 12 months. Figure 12E shows the mean maximum knee extension strength in different dosing regimens. P values ​​<0.05 at 4, 8, and 12 months. There was a 25% decrease at 12 months. Figure 12F shows the maximum elbow flexion strength in different dosing regimens. All p values ​​were NS, no change at 12 months. [Figure 12CD] Same as above. [Figure 12EF] Same as above.

[0030] [Figure 13] Figure 13 depicts self-reported pain scores after treatment with Compound 1. The figure shows the mean ± SEM.

[0031] [Figure 14AB] 14A-14H illustrate the characteristics of short-term and long-term proteomic responses to fast skeletal myosin (eg, Compound 1) in Becker muscular dystrophy (BMD). [Figure 14CD] Same as above. [Figure 14EF] Same as above. [Figure 14GH] Same as above. DETAILED DESCRIPTION OF THE INVENTION

[0032] Detailed Description of the Invention While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various modifications to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby.

[0033] In some embodiments, a method for treating a neuromuscular condition comprises administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day: [ka]

[0013] Methods are described herein that include administering to a subject a compound of Formula (I) at a dose of 15 mg to 40 mg per day. In some embodiments, the methods include administering to a subject a compound of Formula (I) at a dose of 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, or 20 mg per day. In some embodiments, the methods include administering to a subject a compound of Formula (I) at a dose of 10 mg, 15 mg, or 20 mg per day if the subject is older than 18 years of age. In some embodiments, the methods include administering to a subject a compound of Formula (I) at a dose of 10 mg, 7.5 mg, 5 mg, 2.5 mg, 2 mg, or 1 mg per day if the subject is younger than 18 years of age. In some embodiments, administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose. In some embodiments, administering maintains or increases the subject's North Star Ambulatory Assessment (NSAA) score compared to the NSAA score before treatment. In some embodiments, said administering maintains or improves one or more of the following functional measures of the subject: maximum elbow flexion strength compared to maximum elbow flexion strength before treatment; maximum knee extension strength compared to maximum knee extension strength before treatment; 10 meter walking / running speed compared to 10 meter walking / running speed before treatment; 100 meter timed test speed compared to 100 meter timed test speed before treatment; 4-step stair climbing timed speed compared to 4-step stair climbing timed speed before treatment; and maximum grip strength compared to maximum grip strength value before treatment.In some embodiments, said administering maintains creatine kinase activity level or reduces the creatine kinase activity level of the subject by 10% or more compared to the creatine kinase activity level before treatment.In some embodiments, the administering maintains the creatine kinase activity level or increases the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment. In some embodiments, the administering maintains the subject's dual-energy X-ray absorptiometry (DXA) lean mass % or increases the subject's DXA lean mass % by 5% or more compared to the DXA lean mass % before treatment. In some embodiments, the administering maintains the subject's fast skeletal troponin I (TNNI2) concentration or reduces the subject's TNNI2 concentration by 10% or more compared to the TNNI2 concentration before treatment, optionally where the TNNI2 concentration is estimated from SOMAscan levels. In some embodiments, the administering reduces the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more compared to the protein concentration of the one or more muscle injury biomarkers before treatment. In some embodiments, the one or more muscle injury biomarkers include ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof. In some embodiments, the administering reduces the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more pro-inflammatory proteins before treatment. In some embodiments, the one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In some embodiments, the administering increases the protein concentration of the one or more anti-inflammatory proteins in the subject by 10% or more compared to the pre-treatment protein concentration of the one or more anti-inflammatory proteins.In some embodiments, the one or more anti-inflammatory proteins comprise IL10, IL13, IL22, IL37, IL4, or a combination thereof. In some embodiments, the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease. In some embodiments, administering comprises administering a compound of Formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more. In some embodiments, prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a)-(g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a dystrophin gene mutation of -0.1 to -10, or Further comprising selecting subjects for treatment who have one or more of the following: (a) pre-treatment NSAA annual change of -0.5 to -3; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher.

[0034] In certain aspects, the present disclosure provides methods and compositions for treating neuromuscular conditions by selectively inhibiting fast-fiber skeletal muscle myosin. In particular, the methods of the present disclosure can be used in the treatment of DMD, BMD, LGMD, McArdle's disease, and other neuromuscular conditions. In some embodiments, the methods described herein include compounds or salts previously described in the following publications: WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615.

[0035] Skeletal muscles are primarily composed of two types of fibers: slow-twitch fibers (i.e., type I) and fast-twitch fibers (i.e., type II). In each muscle, the two types of fibers are arranged in a mosaic-like arrangement, with differences in fiber type composition in different muscles and at different points in growth and development. Slow-twitch fibers have excellent aerobic energy production capacity. Slow-twitch fibers have a slower contraction rate but are more resistant to fatigue. Slow-twitch fibers typically have higher concentrations of mitochondria and myoglobin than fast-twitch fibers and are surrounded by more capillaries than fast-twitch fibers. Compared to fast-twitch fibers, slow-twitch fibers contract at a slower rate and generate less power due to lower myosin ATPase activity, but they can maintain contractile function for longer periods, such as during stabilization, postural control, and endurance training.

[0036] Human fast-twitch muscle fibers are further divided into two main fiber types depending on the specific fast skeletal myosin they express (type IIa, type IIx / d). A third type of fast fiber (type IIb) is present in other mammals but is rarely identified in human muscles. Fast-twitch muscle fibers have excellent anaerobic energy production capacity and can generate high amounts of tension over short periods of time. Typically, fast-twitch muscle fibers have lower concentrations of mitochondria, myoglobin, and capillaries compared to slow-twitch muscle fibers, and therefore can fatigue more quickly. Fast-twitch muscle fibers produce the force required for power and resistance activities more quickly.

[0037] The proportion of type I and type II fibers can vary among different individuals. For example, a non-athlete may have close to 50% of each muscle fiber type. Power athletes may have a higher proportion of fast-twitch fibers, e.g., 70-75% type II in sprinters. Endurance athletes may have a higher proportion of slow-twitch fibers, e.g., 70-80% in long-distance runners. The proportion of type I and type II fibers can also vary depending on an individual's age. The proportion of type II fibers, especially type IIx, can decline as an individual ages, resulting in a loss of lean muscle mass.

[0038] Contraction of skeletal muscle leads to muscle damage in subjects with neuromuscular diseases, such as DMD, and this damage appears to be more prevalent in fast fibers. It has been observed that the acute force decline after prolonged injury is higher in fast type II fiber muscles compared to slow type I fiber muscles in a dystrophic mouse model. It has also been demonstrated that the degree of acute force decline and histological damage in a dystrophic mouse model is proportional to peak force generation during prolonged injury. Excessive contraction-induced injury precedes the inflammation and irreversible fibrosis that characterize the pathology of end-stage DMD.

[0039] Inhibitors of skeletal muscle myosin that are not selective for type II fibers can result in excessive inhibition of skeletal muscle contraction, including respiratory function, and unwanted inhibition of cardiac activity, since the heart shares some structural components (such as type I myosin) with type I skeletal muscle fibers. Without wishing to be bound by a particular mechanism theory, the present disclosure provides selective inhibitors of fast-twitch fiber skeletal muscle myosin as a treatment option for Becker muscular dystrophy (BMD), Duchenne muscular dystrophy (DMD), limb-girdle muscular dystrophy (LGMD), McArdle disease, and other neuromuscular conditions. Targeted inhibition of type II skeletal muscle myosin can reduce skeletal muscle contraction while minimizing the impact on the subject's daily activities.

[0040] When healthy muscles are subjected to excessive, unaccustomed exercise, pain and persistent reductions in strength and range of motion occur. Proteins, including creatine kinase (CK), lactate dehydrogenase, and myoglobin, also leak into the circulation from injured muscle fibers. These biomarkers are not specific to either fast or slow fibers and therefore do not provide detailed information regarding differences in the fiber response to injury. Troponin I (TNNI) is a component of the troponin complex that regulates the initiation of muscle contraction via calcium. It is distinguished by the existence of different isoforms for each type of striated muscle: TNNI1 in slow skeletal muscles, TNNI2 in fast skeletal muscles, and TNNI3 in cardiac muscles. Selective enzyme-linked immunosorbent assays (ELISAs) have been used to demonstrate that TNNI2, but not TNNI1, is elevated in the circulation after injurious exercise, even under extreme conditions.

[0041] DMD and BMD are caused by the absence (DMD) or shortening (BMD) of dystrophin protein 5. Dystrophin provides a structural link between the actin cytoskeleton and the basement membrane through the dystrophin-glycoprotein complex. When dystrophin is absent or shortened, muscle contraction leads to increased muscle tension and injury with normal use. Although susceptibility to injury is much higher in DMD muscles than in BMD or healthy muscles, fast fibers appear to be more susceptible than slow fibers, with young DMD patients showing histological evidence of destruction of fast fibers 7 and early loss of type IIx fibers. In some embodiments, the present disclosure provides selective inhibitors of fast fiber skeletal muscle myosin as a treatment option for DMD, BMD, McArdle disease, or limb-girdle muscular dystrophy. definition

[0042] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0043] As used in this specification and claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0044] The phrases "parenteral administration" and "administered parenterally", as used herein, mean modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0045] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0046] The phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier," as used herein, refers to a pharmaceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and the like. and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solution; and (21) other non-toxic, compatible substances used in pharmaceutical formulations.

[0047] As used herein, "treatment" or "treating" refers to an approach to obtaining beneficial or desired results for a disease, disorder, or medical condition, including, but not limited to, therapeutic benefit and / or preventative benefit. Therapeutic benefit may include, for example, eradication or amelioration of the underlying disorder being treated. Therapeutic benefit may also include, for example, eradication or amelioration of one or more physiological symptoms associated with the underlying disorder, such that an improvement is observed in a subject, even though the subject may still be suffering from the underlying disorder. In certain embodiments, for preventative benefit, the composition is administered to a subject at risk of developing a particular disease or who reports one or more physiological symptoms of a disease, even though the disease may not have been diagnosed. Treatment via administration of the compounds described herein does not require the involvement of a medical professional.

[0048] The following is a discussion of compounds and their salts that can be used in the methods described herein.In certain embodiments, the compounds or salts of the present disclosure inhibit fast myosin (type II) fast myosin (type II), and are referred to as fast myosin (type II) fast myosin (type II) inhibitors.In certain embodiments, the compounds and salts used in the methods and compositions of the present disclosure are described in any of WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615.In certain embodiments, the compounds used in the methods described herein are compounds of formula (I): [ka] and is also referred to herein as Compound 1. therapeutic application

[0049] In some embodiments, a method for treating a neuromuscular condition comprises administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day:

change

[0050] In some embodiments, the dose of the compound of Formula (I) comprises at least about 1 mg per day to about 30 mg per day. In some embodiments, the dose of the compound of Formula (I) comprises at least about 1 mg per day to about 2 mg per day, about 1 mg per day to about 2.5 mg per day, about 1 mg per day to about 3 mg per day, about 1 mg per day to about 5 mg per day, about 1 mg per day to about 7.5 mg per day, about 1 mg per day to about 10 mg per day, about 1 mg per day to about 12.5 mg per day, about 1 mg per day to about 15 mg per day, about 1 mg per day to about 20 ... Approximately 25mg per day, approximately 1mg per day to approximately 30mg per day, approximately 2mg per day to approximately 2.5mg per day, approximately 2mg per day to approximately 3mg per day, approximately 2mg per day to approximately 5mg per day, approximately 2mg per day to approximately 7.5mg per day, approximately 2mg per day to approximately 10mg per day, approximately 2mg per day to approximately 12.5mg per day, approximately 2mg per day to approximately 15mg per day, approximately 2mg per day to approximately 20mg per day, approximately 2mg per day to approximately 25mg per day, approximately Approximately 2mg to approximately 30mg per day, approximately 2.5mg to approximately 3mg per day, approximately 2.5mg to approximately 5mg per day, approximately 2.5mg to approximately 7.5mg per day, approximately 2.5mg to approximately 10mg per day, approximately 2.5mg to approximately 12.5mg per day, approximately 2.5mg to approximately 15mg per day, approximately 2.5mg to approximately 20mg per day, approximately 2.5mg to approximately 25mg per day, approximately 2.5mg to approximately 30mg per day Approximately 3mg to approximately 5mg per day, approximately 3mg to approximately 7.5mg per day, approximately 3mg to approximately 10mg per day, approximately 3mg to approximately 12.5mg per day, approximately 3mg to approximately 15mg per day, approximately 3mg to approximately 20mg per day, approximately 3mg to approximately 25mg per day, approximately 3mg to approximately 30mg per day, approximately 5mg to approximately 7.5mg per day, approximately 5mg to approximately 10mg per day, approximately 5 ...5mg, approximately 5mg per day to approximately 15mg per day, approximately 5mg per day to approximately 20mg per day, approximately 5mg per day to approximately 25mg per day, approximately 5mg per day to approximately 30mg per day, approximately 7.5mg per day to approximately 10mg per day, approximately 7.5mg per day to approximately 12.5mg per day, approximately 7.5mg per day to approximately 15mg per day, approximately 7.5mg per day to approximately 20mg per day, approximately 7.5mg per day to approximately 25mg per day, approximately 7.5mg per day to approximately 30mg per day, approximately 10mg per day to approximately 12.5mg per day, approximately 10mg per day to approximately 15mg per day, approximately 10mg per day about 20 mg per day, about 10 mg to about 25 mg per day, about 10 mg to about 30 mg per day, about 12.5 mg to about 15 mg per day, about 12.5 mg to about 20 mg per day, about 12.5 mg to about 25 mg per day, about 12.5 mg to about 30 mg per day, about 15 mg to about 20 mg per day, about 15 mg to about 25 mg per day, about 15 mg to about 30 mg per day, about 20 mg to about 25 mg per day, about 20 mg to about 30 mg per day, or about 25 mg to about 30 mg per day. In some embodiments, the dose of the compound of formula (I) is at least about 1 mg per day, about 2 mg per day, about 2.5 mg per day, about 3 mg per day, about 5 mg per day, about 7.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 20 mg per day, about 25 mg per day, or about 30 mg per day. In some embodiments, the dose of the compound of formula (I) is at least about 1 mg per day, about 2 mg per day, about 2.5 mg per day, about 3 mg per day, about 5 mg per day, about 7.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 20 mg per day, or about 25 mg per day. In some embodiments, the dose of the compound of Formula (I) is at least up to about 2 mg per day, about 2.5 mg per day, about 3 mg per day, about 5 mg per day, about 7 mg per day, or about 8 mg per day.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 20 mg per day, about 25 mg per day, or about 30 mg per day.

[0051] In some embodiments, the dose of the compound of Formula (I) includes up to about 1 mg per day to about 30 mg per day. In some embodiments, the dose of the compound of Formula (I) includes up to about 1 mg per day to about 2 mg per day, about 1 mg per day to about 2.5 mg per day, about 1 mg per day to about 3 mg per day, about 1 mg per day to about 5 mg per day, about 1 mg per day to about 7.5 mg per day, about 1 mg per day to about 10 mg per day, about 1 mg per day to about 12.5 mg per day, about 1 mg per day to about 15 mg per day, about 1 mg per day to about 20 mg per day, about 1 mg per day to about 1 Approximately 25mg per day, approximately 1mg per day to approximately 30mg per day, approximately 2mg per day to approximately 2.5mg per day, approximately 2mg per day to approximately 3mg per day, approximately 2mg per day to approximately 5mg per day, approximately 2mg per day to approximately 7.5mg per day, approximately 2mg per day to approximately 10mg per day, approximately 2mg per day to approximately 12.5mg per day, approximately 2mg per day to approximately 15mg per day, approximately 2mg per day to approximately 20mg per day, approximately 2mg per day to approximately 25mg per day, approximately 2mg to approximately 30mg per day, approximately 2.5mg per day to approximately 3mg per day, approximately 2.5mg per day to approximately 5mg per day, approximately 2.5mg per day to approximately 7.5mg per day, approximately 2.5mg per day to approximately 10mg per day, approximately 2.5mg per day to approximately 12.5mg per day, approximately 2.5mg per day to approximately 15mg per day, approximately 2.5mg per day to approximately 20mg per day, approximately 2.5mg per day to approximately 25mg per day, approximately 2.5mg per day to approximately 30mg per day Approximately 3mg to approximately 5mg per day, approximately 3mg to approximately 7.5mg per day, approximately 3mg to approximately 10mg per day, approximately 3mg to approximately 12.5mg per day, approximately 3mg to approximately 15mg per day, approximately 3mg to approximately 20mg per day, approximately 3mg to approximately 25mg per day, approximately 3mg to approximately 30mg per day, approximately 5mg to approximately 7.5mg per day, approximately 5mg to approximately 10mg per day, approximately 5 ...5mg, approximately 5mg per day to approximately 15mg per day, approximately 5mg per day to approximately 20mg per day, approximately 5mg per day to approximately 25mg per day, approximately 5mg per day to approximately 30mg per day, approximately 7.5mg per day to approximately 10mg per day, approximately 7.5mg per day to approximately 12.5mg per day, approximately 7.5mg per day to approximately 15mg per day, approximately 7.5mg per day to approximately 20mg per day, approximately 7.5mg per day to approximately 25mg per day, approximately 7.5mg per day to approximately 30mg per day, approximately 10mg per day to approximately 12.5mg per day, approximately 10mg per day to approximately 15mg per day, approximately 10mg per day about 20 mg per day, about 10 mg to about 25 mg per day, about 10 mg to about 30 mg per day, about 12.5 mg to about 15 mg per day, about 12.5 mg to about 20 mg per day, about 12.5 mg to about 25 mg per day, about 12.5 mg to about 30 mg per day, about 15 mg to about 20 mg per day, about 15 mg to about 25 mg per day, about 15 mg to about 30 mg per day, about 20 mg to about 25 mg per day, about 20 mg to about 30 mg per day, or about 25 mg to about 30 mg per day. In some embodiments, the dose of the compound of Formula (I) comprises up to about 1 mg per day, about 2 mg per day, about 2.5 mg per day, about 3 mg per day, about 5 mg per day, about 7.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 20 mg per day, about 25 mg per day, or about 30 mg per day. In some embodiments, the dose of the compound of Formula (I) comprises up to at least about 1 mg per day, about 2 mg per day, about 2.5 mg per day, about 3 mg per day, about 5 mg per day, about 7.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 20 mg per day, or about 25 mg per day. In some embodiments, the dose of the compound of Formula (I) is at most about 2 mg per day, at most about 2.5 mg per day, at most about 3 mg per day, at most about 5 mg per day, at most about 7.5 mg per day, at most about 10 mg per day, at most about 12 mg per day.5 mg per day, about 15 mg per day, about 20 mg per day, about 25 mg per day, or about 30 mg per day.

[0052] In some embodiments, administering to the subject comprises a 20 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 1 mg to 20 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, or 20 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 1 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 2 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 2.5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 7.5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to a subject comprises a 10 mg dose of a compound of Formula (I) per day. In some embodiments, administering to a subject comprises a 15 mg dose of a compound of Formula (I) per day. In some embodiments, administering to a subject comprises a 20 mg dose of a compound of Formula (I) per day.

[0053] In some embodiments, the subject treated with the compound of Formula (I) is over 18 years of age. In some embodiments, administering to the subject comprises a 10 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 15 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 20 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 10 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 7.5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 2.5 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 2 mg dose of the compound of Formula (I) per day. In some embodiments, administering to the subject comprises a 1 mg dose of the compound of Formula (I) per day. In some embodiments, administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose. In some embodiments, administering maintains or increases the subject's North Star Ambulatory Assessment (NSAA) score compared to the NSAA score before treatment. In some embodiments, the administration maintains or improves one or more of the following functional measures in the subject: maximum elbow flexion strength compared to maximum elbow flexion strength before treatment; maximum knee extension strength compared to maximum knee extension strength before treatment; 10 meter walk / run speed compared to 10 meter walk / run speed before treatment; 100 meter timed test speed compared to 100 meter timed test speed before treatment; 4-step stair climbing timed speed compared to 4-step stair climbing timed speed before treatment; and maximum grip strength compared to maximum grip strength value before treatment.In some embodiments, the administering maintains the creatine kinase activity level or reduces the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment. In some embodiments, the administering maintains the creatine kinase activity level or increases the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment. In some embodiments, the administering maintains the subject's dual-energy X-ray absorptiometry (DXA) lean mass % or increases the subject's DXA lean mass % by 5% or more compared to the DXA lean mass % before treatment. In some embodiments, the administering maintains the subject's fast skeletal troponin I (TNNI2) concentration or reduces the subject's TNNI2 concentration by 10% or more compared to the TNNI2 concentration before treatment, optionally where the TNNI2 concentration is estimated from SOMAscan levels. In some embodiments, the administering reduces the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more compared to the pre-treatment protein concentration of the one or more muscle injury biomarkers. In some embodiments, the one or more muscle injury biomarkers include ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof. In some embodiments, the administering reduces the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more pro-inflammatory proteins before treatment. In some embodiments, the one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof.In some embodiments, the administering increases the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more anti-inflammatory proteins before treatment. In some embodiments, the one or more anti-inflammatory proteins include IL10, IL13, IL22, IL37, IL4, or a combination thereof. In some embodiments, the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease. In some embodiments, the administering comprises administering a compound of Formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more. In some embodiments, prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a)-(g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a dystrophin gene mutation of -0.1 to -10, or Further comprising selecting subjects for treatment who have one or more of the following: (a) pre-treatment NSAA annual change of -0.5 to -3; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher.

[0054] The methods described herein are useful for treating neuromuscular conditions. In some embodiments, the methods include administering a composition comprising a fast-twitch myosin (type II) inhibitor and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents, and / or thickeners. In some embodiments, the fast-twitch myosin (type II) inhibitor is a compound described in WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, WO2021231615, or PCT application PCT / US2023 / 021739. In some embodiments, fast twitch myosin (type II) inhibitors are described in WO2020097266. In some embodiments, the fast twitch myosin (type II) inhibitor is represented by Formula (I), also referred to as Compound 1: [ka] It is expressed as:

[0055] The administration method of Compound 1 or other fast-twitch myosin (type II) inhibitors can be used to treat neuromuscular conditions and movement disorders.Examples of neuromuscular conditions include, but are not limited to, Duchenne muscular dystrophy, Becker muscular dystrophy, myotonic dystrophy 1, myotonic dystrophy 2, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, limb-girdle muscular dystrophy, tendonitis, and carpal tunnel syndrome.Examples of movement disorders include, but are not limited to, muscle spasticity disorder, spasticity associated with multiple sclerosis, Parkinson's disease, Alzheimer's disease, or cerebral palsy, or injury or traumatic event, such as stroke, traumatic brain injury, spinal cord injury, hypoxia, meningitis, encephalitis, phenylketonuria, or amyotrophic lateral sclerosis. Other conditions that may respond to inhibition of skeletal myosin II, skeletal troponin C, skeletal troponin I, skeletal tropomyosin, skeletal troponin T, skeletal regulatory light chain, skeletal myosin-binding protein C, or skeletal actin are also included. In some embodiments, the neuromuscular condition and movement disorder is selected from muscular dystrophies and myopathies. In some embodiments, muscular dystrophies are diseases in which abnormal genes (mutations) prevent the production of proteins necessary for healthy muscle formation, causing progressive weakness and loss of muscle mass. In some embodiments, the muscular dystrophies are selected from Becker muscular dystrophy (BMD), congenital muscular dystrophy (CMD), Duchenne muscular dystrophy (DMD), Emery-Dreifuss muscular dystrophy (EDMD), facioscapulohumeral muscular dystrophy (FSHD), limb-girdle muscular dystrophy (LGMD), myotonic dystrophy (DM), and oculopharyngeal muscular dystrophy (OPMD). In some embodiments, the congenital muscular dystrophy (CMD) is selected from Bethlem CMD, Fukuyama CMD, muscle-eye-brain disease (MEB), ankylosing spine syndrome, Ullrich CMD, and Walker-Warburg syndrome (WWS). In some embodiments, the myopathy is a muscle disease not caused by neuropathy. Myopathy causes muscles to weaken or shrink (atrophy).In some embodiments, the myopathy is selected from congenital myopathy, distal myopathy, endocrine myopathy, inflammatory myopathy, metabolic myopathy, myofibrillar myopathy (MFM), scapuloperoneal myopathy, and cardiomyopathies. In some embodiments, the congenital myopathy is selected from cap myopathy, centronuclear myopathy, congenital myopathy with fiber type disproportion, core myopathy, central core disease, multiminicore myopathy, myosin storage myopathy, myotubular myopathy, and nemaline myopathy. In some embodiments, the distal myopathy is selected from GNE myopathy / Nonaka myopathy / hereditary inclusion body myopathy (HIBM), Laing distal myopathy, Marquesberg-Griggs late-onset distal myopathy, Miyoshi myopathy, Udd myopathy / tibial muscular dystrophy, VCP myopathy / IBMPFD, distal vocal cord and pharyngeal myopathy, and Welander distal myopathy. In some embodiments, the endocrine myopathy is selected from hyperthyroid myopathy and hypothyroid myopathy. In some embodiments, the inflammatory myopathy is selected from dermatomyositis, inclusion body myositis, and polymyositis. In some embodiments, the metabolic myopathy is selected from von Gierke disease, Anderson disease, Fanconi-Bickel syndrome, aldolase A deficiency, acid maltase deficiency (Pompe disease), carnitine deficiency, carnitine palmitoyltransferase deficiency, debranching enzyme deficiency (Cori disease, Forbes disease), lactate dehydrogenase deficiency, myoadenylate deaminase deficiency, phosphofructokinase deficiency (Tarui disease), phosphoglycerate kinase deficiency, phosphoglycerate mutase deficiency (e.g., glycogen storage disease X), and phosphorylase deficiency (McArdle disease). In some embodiments, the cardiomyopathy is selected from intrinsic cardiomyopathy and extrinsic cardiomyopathy. In some embodiments, the intrinsic cardiomyopathy is selected from genetic myopathy and acquired myopathy. In some embodiments, the genetic myopathy is selected from hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy (ARVC), LV noncompaction, ion channelopathy, dilated cardiomyopathy (DCM), and restrictive cardiomyopathy (RCM).In some embodiments, the acquired myopathy is selected from stress cardiomyopathy, myocarditis, eosinophilic myocarditis, and ischemic cardiomyopathy. In some embodiments, the exogenous cardiomyopathy is selected from metabolic cardiomyopathy, endomyocardial cardiomyopathy, endocrine cardiomyopathy, and cardiofacial cardiomyopathy. In some embodiments, the metabolic cardiomyopathy is selected from Fabry disease and hemochromatosis. In some embodiments, the endomyocardial cardiomyopathy is selected from endomyocardial fibrosis and hypereosinophilic syndrome. In some embodiments, the endocrine cardiomyopathy is selected from diabetes mellitus, hyperthyroidism, and acromegaly. In some embodiments, the cardiofacial cardiomyopathy is Noonan syndrome.

[0056] In some aspects, methods for treating a neuromuscular condition or movement disorder may include administering Compound 1 or a salt thereof to inhibit skeletal muscle contraction. In some embodiments, Compound 1 or a salt thereof does not significantly inhibit myocardial contraction. In some embodiments, myocardial contraction is inhibited by 20% or less. In some embodiments, myocardial contraction is inhibited by 15% or less. In some embodiments, myocardial contraction is inhibited by 10% or less. In some embodiments, myocardial contraction is inhibited by 9% or less. In some embodiments, myocardial contraction is inhibited by 8% or less. In some embodiments, myocardial contraction is inhibited by 7% or less. In some embodiments, myocardial contraction is inhibited by 6% or less. In some embodiments, myocardial contraction is inhibited by 5% or less. In some embodiments, myocardial contraction is inhibited by 4% or less. In some embodiments, myocardial contraction is inhibited by 3% or less. In some embodiments, myocardial contraction is inhibited by 2% or less. In some embodiments, myocardial contraction is inhibited by 1% or less.

[0057] The activities of daily living (ADL) or habitual physical activity of the subject can be monitored before and after treatment with compound 1 or its salt.ADL or habitual physical activity depends on the subject, and can range from simple walking to extensive exercise according to the subject's ability and routine.The treatment options and dosage of the skeletal muscle contraction inhibitor discussed herein can be individualized for the subject, so that ADL and habitual physical activity remain unchanged.

[0058] In some aspects, a method for treating a neuromuscular condition or movement disorder can include administering Compound 1 or a salt thereof to inhibit skeletal muscle contraction. Compound 1 or a salt thereof can be administered in an amount relative to the amount required to reduce skeletal muscle contraction by 50%. Compound 1 or a salt thereof can be administered in an amount less than the amount required to reduce skeletal muscle contraction by 50% compared to the subject's skeletal muscle contractile capacity before treatment. Compound 1 or a salt thereof can be administered in an amount that reduces skeletal muscle contraction by 5% to 45% compared to the subject's skeletal muscle contractile capacity before treatment. In some cases, Compound 1 or a salt thereof can be administered in an amount that reduces skeletal muscle contraction by less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, less than 35%, less than 40%, less than 45%, or even less than 50% compared to the subject's skeletal muscle contractile capacity before treatment. In certain embodiments, Compound 1 or a salt thereof can be administered in an amount that reduces skeletal muscle contraction by 1% to 50% compared to the subject's skeletal muscle contractile capacity before treatment.

[0059] In some embodiments, a method for treating a neuromuscular condition or movement disorder may include administering Compound 1 or a salt thereof to inhibit type I skeletal muscle contraction. The inhibitor of type I skeletal muscle contraction may be administered in an amount relative to the amount required to reduce type I skeletal muscle contraction by 20%. The inhibitor of type I skeletal muscle contraction may be administered in an amount less than the amount required to reduce type I skeletal muscle contraction by 20% compared to the subject's type I skeletal muscle contraction capacity before treatment. The inhibitor of type I skeletal muscle contraction may be administered in an amount that reduces type I skeletal muscle contraction by 0.01% to 20% compared to the subject's type I skeletal muscle contraction capacity before treatment. In some cases, the inhibitor may be administered in an amount that reduces type I skeletal muscle contraction by less than 0.01%, less than 0.1%, less than 0.5%, less than 1%, less than 5%, less than 10%, less than 15%, or less than 20% compared to the subject's type I skeletal muscle contraction capacity before treatment. In certain embodiments, the inhibitor may be administered in an amount that reduces type I skeletal muscle contraction by 0.01% to 20% compared to the subject's pre-treatment type I skeletal muscle contraction capacity.

[0060] In some embodiments, a method for treating a neuromuscular condition or movement disorder may include administering Compound 1 or a salt thereof to inhibit type II skeletal muscle contraction. The inhibitor of type II skeletal muscle contraction may be administered in an amount relative to the amount required to reduce type II skeletal muscle contraction by 90%. The inhibitor of type II skeletal muscle contraction may be administered in an amount less than the amount required to reduce type II skeletal muscle contraction by 90% compared to the subject's type II skeletal muscle contraction capacity before treatment. The inhibitor of type II skeletal muscle contraction may be administered in an amount that reduces type II skeletal muscle contraction by 5% to 75% compared to the subject's type II skeletal muscle contraction capacity before treatment. In some cases, the inhibitor may be administered in an amount that reduces type II skeletal muscle contraction by less than 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, or even less than 90% compared to the subject's type II skeletal muscle contraction capacity before treatment. In certain embodiments, the inhibitor may be administered in an amount that reduces type II skeletal muscle contraction by 1% to 50% compared to the subject's type II skeletal muscle contraction capacity before treatment.

[0061] In some aspects, methods for treating contraction-induced injury in skeletal muscle fibers can include administering Compound 1 or a salt thereof to inhibit skeletal muscle contraction and / or fast skeletal myosin (type II). In certain embodiments, the inhibitor does not appreciably inhibit myocardial contraction.

[0062] In some embodiments, a method of treating a metabolic myopathy, such as McArdle syndrome, can include administering Compound 1 or a salt thereof.

[0063] In certain embodiments, contraction-induced damage in skeletal muscle fibers is due to involuntary skeletal muscle contractions. Involuntary skeletal muscle contractions may be associated with neuromuscular conditions or spasticity-related conditions. In certain embodiments, contraction-induced damage in skeletal muscle fibers is due to voluntary skeletal muscle contractions, such as physical exercise.

[0064] In certain embodiments, administration of Compound 1 or a salt thereof to a subject modulates one or more biomarkers related to muscle contraction. Examples of biomarkers include, but are not limited to, creatinine, creatine kinase (CK), troponin T (TnT), troponin C (TnC), troponin I (TnI), pyruvate kinase (PK), lactate dehydrogenase (LDH), myoglobin, isoforms of TnI (cardiac, slow skeletal muscle, fast skeletal muscle, etc.), and inflammatory markers (IL-1, IL-6, IL-4, TNF-α). Biomarkers may also include measures of muscle inflammation, such as edema. The levels of the biomarkers described herein may increase after administration of an inhibitor compared to the levels of the biomarker before treatment. Alternatively, the levels of the biomarkers may decrease after administration of an inhibitor compared to the levels of the biomarker before treatment. Modulation of one or more biomarkers by an inhibitor described herein may indicate treatment of a neuromuscular condition, such as those described herein.

[0065] In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more muscle injury biomarkers (e.g., one or more of the muscle injury proteins shown in Figure 14D). Examples of the one or more muscle injury biomarkers may include ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof. In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more muscle injury biomarkers during short-term treatment (e.g., 1-2 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more biomarkers of muscle injury during medium-term treatment (e.g., 3-4 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more biomarkers of muscle injury during long-term treatment (e.g., 6-12 months of treatment with Compound 1).

[0066] In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins (e.g., one or more of the pro-inflammatory proteins shown in Figure 14G or Figure 14H). Examples of the one or more pro-inflammatory proteins may include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during short-term treatment (e.g., 1-2 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during medium-term treatment (e.g., 3-4 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during long-term treatment (e.g., 6-12 months of treatment with Compound 1).

[0067] In certain embodiments, administration of Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins (e.g., one or more of the anti-inflammatory proteins shown in Figure 14G or Figure 14H). Examples of the one or more anti-inflammatory proteins may include IL10, IL13, IL22, IL37, IL4, or a combination thereof. In certain embodiments, administration of Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during short-term treatment (e.g., 1-2 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during medium-term treatment (e.g., 3-4 months of treatment with Compound 1). In certain embodiments, administration of Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during long-term treatment (e.g., 6-12 months of treatment with Compound 1).

[0068] The level of CK in a subject increases when the subject is active compared to when the subject is inactive (for example, during sleep), and therefore CK is a potential metric for evaluating skeletal muscle destruction caused by skeletal muscle contraction.In certain embodiments, compound 1 or its salt can be administered to a subject before light, moderate, or vigorous activity to reduce or prevent skeletal muscle destruction from activity.Moderate to vigorous activity can depend on the subject's ability, and can include physical exercise that increases the subject's heart rate by at least 20% or more, for example, about 50% or more, compared to the subject's resting heart rate.Examples of moderate to vigorous activity include walking, running, weightlifting, cycling, swimming, hiking, etc.

[0069] In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, is administered to a subject in an amount sufficient to maintain creatine kinase activity levels or reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, the subject's creatine kinase activity level is assessed after the first dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the assessment is performed 1 month or more, 2 months or more, 3 months or more, 4 months or more, 6 months or more, 8 months or more, 12 months or more, 24 months or more, 18 months or more, or 24 months or more. In some embodiments, the subject's creatine kinase activity level is assessed after the first dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the assessment is performed 1 month or more, 2 months or more, 3 months or more, 4 months or more, 6 months or more, 8 months or more, 12 months or more, 24 months or more, 18 months or more, or 24 months or more after the first dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor. In some embodiments, the one or more subsequent doses are higher than the first dose. In some embodiments, the one or more subsequent doses are lower than the first dose. In some embodiments, the one or more subsequent doses are determined based on the level of creatine kinase activity assessed after the first dose of the fast-twitch myosin (type II) inhibitor to the subject.In some embodiments, one or more subsequent doses are determined based on the level of creatine kinase activity assessed after a previous dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the pre-treatment creatine kinase activity level is assessed within one month before the first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0070] In certain embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered before, during, or after moderate or vigorous activity to reduce or prevent skeletal muscle breakdown from the activity. Compound 1 or a salt thereof can reduce the subject's CK levels compared to untreated subjects performing the same activity. CK levels can be measured in the subject's peripheral blood during or after the activity. Administration of an inhibitor described herein can reduce CK levels by 5% to 90% in an active subject compared to untreated subjects performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of an inhibitor described herein can modulate CK levels by about 5% to about 90% compared to untreated subjects performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of an inhibitor described herein can reduce CK levels by at least about 5% compared to untreated subjects performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of the inhibitors described herein can modulate CK levels by up to about 90% compared to untreated subjects performing the same activity.Administration of the inhibitors described herein may increase CK levels by about 5% to about 15%, about 5% to about 25%, about 5% to about 35%, about 5% to about 45%, about 5% to about 55%, about 5% to about 65%, about 5% to about 75%, about 5% to about 85%, about 5% to about 90%, about 15% to about 25%, about 15% to about 35%, about 15% to about 45%, about 15% to about 55%, about 15% to about 65%, about 15% to about 75%, about 15% to about 85%, about 15% to about 90%, about 25% to about 35%, about 25% to about 45%, about 25% to about 55%, about 25% to about 65%, about 25% to about 75%, about 25% to about 85%, about 25% to about 90%, about 35% to about 45%, about 35% to about 55%, about 35% to about 65%, about 35% to about 75%, about 35% to about 85%, about 3 ... The skeletal muscle breakdown from activity may be reduced by about 85%, about 25% to about 90%, about 35% to about 45%, about 35% to about 55%, about 35% to about 65%, about 35% to about 75%, about 35% to about 85%, about 35% to about 90%, about 45% to about 55%, about 45% to about 65%, about 45% to about 75%, about 45% to about 85%, about 45% to about 90%, about 55% to about 65%, about 55% to about 75%, about 55% to about 85%, about 55% to about 90%, about 65% to about 75%, about 65% to about 85%, about 65% to about 90%, about 75% to about 85%, about 75% to about 90%, or about 85% to about 90%, thereby reducing or preventing skeletal muscle breakdown from activity. Administration of the inhibitors described herein may modulate CK levels by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90% compared to untreated subjects performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity.

[0071] Treatment with a fast-twitch myosin (type II) inhibitor, such as Compound 1, sustained for various periods can reduce a subject's creatine kinase activity level. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for one month to maintain the creatine kinase activity level or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for two months to maintain the creatine kinase activity level or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for three months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for four months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment.In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 5 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 6 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 7 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 8 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment.In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 9 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 10 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 11 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 12 months to maintain creatine kinase activity levels or to reduce the subject's creatine kinase activity level by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more compared to the creatine kinase activity level before treatment.

[0072] In certain embodiments, the fast-twitch myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or increase the subject's lean mass as determined by dual-energy X-ray absorptiometry (DXA) %. In some embodiments, the subject's lean mass is increased by 5% or more, 10% or more, 15% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, the subject's lean mass is assessed after the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the assessment is performed 1 month or more, 2 months or more, 3 months or more, 4 months or more, 6 months or more, 8 months or more, 12 months or more, 24 months or more, 18 months or more, or 24 months or more after the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor. In some embodiments, the one or more subsequent doses are higher than the first dose. In some embodiments, the one or more subsequent doses are lower than the first dose. In some embodiments, the one or more subsequent doses are determined based on the level of lean mass assessed after the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, one or more subsequent doses are determined based on the level of lean mass assessed after one or more previous doses of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the pre-treatment lean mass level is assessed within one month prior to the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject.

[0073] Administration of Compound 1 or a salt thereof to a subject can modulate the level of circulating fast skeletal troponin I (fS-TnI or TNNI2). The level of fS-TnI can be measured in peripheral blood. The level of fS-TnI in peripheral blood can be increased after administration of the inhibitor compared to the level of fS-TnI in the subject before treatment. Alternatively, the level of fS-TnI in peripheral blood can be decreased after administration of the inhibitor compared to the level of fS-TnI in the subject before treatment. Administration of an inhibitor described herein can modulate the level of fS-TnI by 5% to 90% compared to the level of fS-TnI in the subject before treatment. In some cases, the level of fS-TnI can be modulated by at least about 5% compared to the level of fS-TnI in the subject before treatment. In some cases, the level of fS-TnI can be modulated by up to about 90% compared to the level of fS-TnI in the subject before treatment. In some cases, the level of fS-TnI is about 5% to about 15%, about 5% to about 25%, about 5% to about 35%, about 5% to about 45%, about 5% to about 55%, about 5% to about 65%, about 5% to about 75%, about 5% to about 85%, about 5% to about 90%, about 15% to about 25%, about 15% to about 35%, about 15% to about 45%, about 15% to about 55%, about 15% to about 65%, about 15% to about 75%, about 15% to about 85%, about 15% to about 90%, about 25% to about 35%, about 25% to about 45%, about 25% to about 55%, about 25% to about 65%, about 25% to about 90%, or It may be modulated by about 75%, about 25% to about 85%, about 25% to about 90%, about 35% to about 45%, about 35% to about 55%, about 35% to about 65%, about 35% to about 75%, about 35% to about 85%, about 35% to about 90%, about 45% to about 55%, about 45% to about 65%, about 45% to about 75%, about 45% to about 85%, about 45% to about 90%, about 55% to about 65%, about 55% to about 75%, about 55% to about 85%, about 55% to about 90%, about 65% to about 75%, about 65% to about 85%, about 65% to about 90%, about 75% to about 85%, about 75% to about 90%, or about 85% to about 90%.In some cases, the level of fS-TnI may be modulated by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90% compared to the subject's level of fS-TnI before treatment.

[0074] Troponin isoforms can be measured in subjects before and after administration of Compound 1 or a salt thereof. Inhibition of skeletal muscle contraction may not inhibit some isoforms of troponin, such as cardiac troponin I (cTnI) or slow skeletal troponin I (ssTnI). In some cases, inhibition of skeletal muscle contraction may not appreciably inhibit cTnI or ssTnI. When used herein with respect to cTnI or ssTnI, this phrase refers to cTnI or ssTnI that is not appreciably reduced by less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1%, less than 0.5%, or even less than 0.1% compared to cTnI or ssTnI before administration of the inhibitor.

[0075] In certain embodiments, the fast myosin (type II) inhibitor, for example, Compound 1 or its salt, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration of the subject compared to the TNNI2 concentration before treatment.In some embodiments, the TNNI2 of the subject is reduced by 5% or more, 10%, 15%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or more.In some embodiments, the TNNI2 of the subject is evaluated after the first dose of the fast myosin (type II) inhibitor to the subject.In some embodiments, the evaluation is carried out 1 month or longer, 2 months or longer, 3 months or longer, 4 months or longer, 6 months or longer, 8 months or longer, 12 months or longer, 24 months or longer, 18 months or longer, or 24 months or longer after the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor. In some embodiments, the one or more subsequent doses are higher than the first dose. In some embodiments, the one or more subsequent doses are lower than the first dose. In some embodiments, the one or more subsequent doses are determined based on the TNNI2 level assessed after the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the one or more subsequent doses are determined based on the TNNI2 level assessed after one or more previous doses of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the pre-treatment TNNI2 level is assessed within one month before the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject.

[0076] The treatment of fast myosin (type II) inhibitor, for example, compound 1, that continues for various periods can reduce the TNNI2 level of the subject.In some embodiments, fast myosin (type II) inhibitor, for example, compound 1, is administered to the subject for one month to maintain TNNI2 level, or to reduce the TNNI2 level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 level before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for two months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 3 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment.In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 4 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 5 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 6 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment.In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 7 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 8 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 9 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment.In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 10 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 11 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment. In some embodiments, a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 12 months to maintain TNNI2 levels or to reduce the subject's TNNI2 levels by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, 80% or more, or 90% or more compared to the TNNI2 levels before treatment.

[0077] Because elevated serum myoglobin levels can be found in patients with some types of neuromuscular conditions, myoglobin can be a potential metric for assessing the progression of neuromuscular conditions. In some embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1, is administered to a subject in an amount sufficient to maintain or reduce the subject's myoglobin level by 5% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, 40% or more, 45% or more, or 50% or more compared to the myoglobin level before treatment. In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, maintains myoglobin levels or increases a subject's myoglobin levels by about 5% to about 15%, about 5% to about 25%, about 5% to about 35%, about 5% to about 45%, about 5% to about 55%, about 5% to about 65%, about 5% to about 75%, about 5% to about 85%, about 5% to about 90%, about 15% to about 25%, about 15% to about 35%, about 15% to about 45%, about 15% to about 55%, about 15% to about 65%, about 15% to about 75%, about 15% to about 85%, about 15% to about 90%, about 25% to about 35%, about 25% to about 45%, about 25% to about 5 ... 5%, about 25% to about 65%, about 25% to about 75%, about 25% to about 85%, about 25% to about 90%, about 35% to about 45%, about 35% to about 55%, about 35% to about 65%, about 35% to about 75%, about 35% to about 85%, about 35% to about 90%, about 45% to about 55%, about 45% to about 65%, about 45% to about 75%, about 45% The subject is administered an amount sufficient to reduce the level of vasoconstriction by about 85%, about 45% to about 90%, about 55% to about 65%, about 55% to about 75%, about 55% to about 85%, about 55% to about 90%, about 65% to about 75%, about 65% to about 85%, about 65% to about 90%, about 75% to about 85%, about 75% to about 90%, or about 85% to about 90%.In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, is administered to a subject in an amount sufficient to maintain myoglobin levels or to reduce the subject's myoglobin levels by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90% compared to the myoglobin levels before treatment.

[0078] In some embodiments, the subject's myoglobin level is assessed after the first dose of the fast-twitch myosin (type II) inhibitor is administered to the subject. In some embodiments, the assessment is performed 1 month or more, 2 months or more, 3 months or more, 4 months or more, 6 months or more, 8 months or more, 12 months or more, 24 months or more, 18 months or more, or 24 months or more after the first dose of the fast-twitch myosin (type II) inhibitor is administered to the subject. In some embodiments, the administering step comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor. In some embodiments, the one or more subsequent doses are higher than the first dose. In some embodiments, the one or more subsequent doses are lower than the first dose. In some embodiments, one or more subsequent doses are determined based on the myoglobin level assessed after a first dose of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, one or more subsequent doses are determined based on the myoglobin level assessed after one or more previous doses of the fast-twitch myosin (type II) inhibitor administered to the subject. In some embodiments, the pre-treatment myoglobin level is assessed within one month before the first dose of the fast-twitch myosin (type II) inhibitor administered to the subject.

[0079] In certain embodiments, a fast-twitch myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered before, during, or after moderate or vigorous activity to reduce or prevent skeletal muscle breakdown from the activity. Compound 1 or a salt thereof can reduce the subject's myoglobin level compared to an untreated subject performing the same activity. The myoglobin level can be measured in the subject's peripheral blood during or after the activity. Administration of an inhibitor described herein can reduce the myoglobin level in an active subject by 5% to 90% compared to an untreated subject performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of an inhibitor described herein can modulate the myoglobin level by about 5% to about 90% compared to an untreated subject performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of an inhibitor described herein can reduce the myoglobin level by at least about 5% compared to an untreated subject performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity. Administration of the inhibitors described herein can modulate myoglobin levels by up to about 90% compared to untreated subjects performing the same activity.Administration of the inhibitors described herein may increase myoglobin levels by about 5% to about 15%, about 5% to about 25%, about 5% to about 35%, about 5% to about 45%, about 5% to about 55%, about 5% to about 65%, about 5% to about 75%, about 5% to about 85%, about 5% to about 90%, about 15% to about 25%, about 15% to about 35%, about 15% to about 45%, about 15% to about 55%, about 15% to about 65%, about 15% to about 75%, about 15% to about 85%, about 15% to about 90%, about 25% to about 35%, about 25% to about 45%, about 25% to about 55%, about 25% to about 65%, about 25% to about 75%, about 25% to about 85%, about 25% to about 90%, or about 35% to about 45% compared to an untreated subject performing the same activity. % to about 85%, about 25% to about 90%, about 35% to about 45%, about 35% to about 55%, about 35% to about 65%, about 35% to about 75%, about 35% to about 85%, about 35% to about 90%, about 45% to about 55%, about 45% to about 65%, about 45% to about 75%, about 45% to about 85%, about 45% to about 90%, about 55% to about 65%, about 55% to about 75%, about 55% to about 85%, about 55% to about 90%, about 65% to about 75%, about 65% to about 85%, about 65% to about 90%, about 75% to about 85%, about 75% to about 90%, or about 85% to about 90%, thereby reducing or preventing skeletal muscle breakdown from activity. Administration of the inhibitors described herein may modulate myoglobin levels by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90% compared to untreated subjects performing the same activity, thereby reducing or preventing skeletal muscle breakdown from the activity.

[0080] In certain embodiments, fast myosin (type II) inhibitor, for example, compound 1 or its salt, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration of the target compared with the TNNI2 concentration before treatment.In some embodiments, TNNI2 concentration is reduced by 120ng / mL or more, 100ng / mL or more, 80ng / mL or more, 65ng / mL or more, 50ng / mL or more, 45ng / mL or more, 40ng / mL or more, 35ng / mL or more, 30ng / mL or more, 25ng / mL or more, 20ng / mL or more, 15ng / mL or more, 10ng / mL or more, 5ng / mL or more. In some embodiments, the TNNI2 concentration is reduced by between about 5 and 120 ng / mL, between about 10 and 120 ng / mL, between about 20 and 100 ng / mL, between 5 and 50 ng / mL, between about 5 and 45 ng / mL, between about 5 and 40 ng / mL, between about 5 and 35 ng / mL, between about 5 and 30 ng / mL, between about 5 and 25 ng / mL, between about 5 and 20 ng / mL, between about 5 and 15 ng / mL, between about 5 and 10 ng / mL, between about 10 and 40 ng / mL, between about 20 and 40 ng / mL, between about 25 and 35 ng / mL, between about 15 and 35 ng / mL, between about 10 and 20 ng / mL, between about 40 and 50 ng / mL, or between about 20 and 35 ng / mL.

[0081] In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's TNNI2 concentration to about 1 ng / mL to about 50 ng / mL. In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's TNNI2 concentration to about 1 ng / mL to about 3 ng / mL, about 1 ng / mL to about 5 ng / mL, about 1 ng / mL to about 8 ng / mL, about 1 ng / mL to about 10 ng / mL, about 1 ng / mL to about 15 ng / mL, about 1 ng / mL to about 20 ng / mL, about 1 ng / mL to about 25 ng / mL, about 1 ng / mL to about 30 ng / mL, about 1 ng / mL to about 35 ng / mL, about 1 ng / mL to about 40 ng / mL, about 1 ng / mL to about 50 ng / mL. ng / mL~about 50ng / mL, about 3ng / mL~about 5ng / mL, about 3ng / mL~about 8ng / mL, about 3ng / mL~about 10ng / mL, about 3ng / mL~about 15ng / mL, about 3ng / mL~about 20ng / mL, about 3ng / mL~about 25ng / mL, about 3ng / mL to about 30ng / mL, about 3ng / mL to about 35ng / mL, about 3ng / mL to about 40ng / mL, about 3ng / mL to about 50ng / mL, about 5ng / mL to about 8ng / mL, about 5ng / mL to about 10ng / mL, about 5ng / mL ~15ng / mL, 5ng / mL~20ng / mL, 5ng / mL~25ng / mL, 5ng / mL~30ng / mL, 5ng / mL~35ng / mL, 5ng / mL~40ng / mL, 5ng / mL~50ng / mL , about 8ng / mL to about 10ng / mL, about 8ng / mL to about 15ng / mL, about 8ng / mL to about 20ng / mL, about 8ng / mL to about 25ng / mL, about 8ng / mL to about 30ng / mL, about 8ng / mL to about 35ng / mL, about 8ng / mL to about 40ng / mL, about 8ng / mL to about 50ng / mL, about 10ng / mL to about 15ng / mL, about 10ng / mL to about 20ng / mL, about 10ng / mL to about 25ng / mL, about 10ng / mL to about 30ng / mL, about 10ng / mL to about 35ng / mL, about 10ng / mL to about 40ng / mL, about 10ng / mL to about 50ng / mL, about 15ng / mL to about 20ng / mL, about 15ng / mL to about 25ng / mL, about 15ng / mL to about 30ng / mL, about 15ng / mL to about 35ng / mL,Approximately 15ng / mL to approximately 40ng / mL, approximately 15ng / mL to approximately 50ng / mL, approximately 20ng / mL to approximately 25ng / mL, approximately 20ng / mL to approximately 30ng / mL, approximately 20ng / mL to approximately 35ng / mL, about 20ng / mL to about 40ng / mL, about 20ng / mL to about 50ng / mL, about 25ng / mL to about 30ng / mL, about 25ng / mL to about 35ng / mL, about 25 In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's TNNI2 concentration to about 1 ng / mL, about 3 ng / mL, about 5 ng / mL, about 8 ng / mL, about 10 ng / mL, about 15 ng / mL, about 20 ng / mL, about 25 ng / mL, about 30 ng / mL, about 35 ng / mL, about 30 ng / mL, about 40 ng / mL, about 30 ng / mL, about 50 ng / mL, about 35 ng / mL, about 40 ng / mL, or about 50 ng / mL. In certain embodiments, the fast myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's TNNI2 concentration to at least about 1 ng / mL, about 3 ng / mL, about 5 ng / mL, about 8 ng / mL, about 10 ng / mL, about 15 ng / mL, about 20 ng / mL, about 25 ng / mL, about 30 ng / mL, about 35 ng / mL, or about 40 ng / mL. In certain embodiments, the fast myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's TNNI2 concentration to at most about 3 ng / mL, about 5 ng / mL, about 8 ng / mL, about 10 ng / mL, about 15 ng / mL, about 20 ng / mL, about 25 ng / mL, about 30 ng / mL, about 35 ng / mL, about 40 ng / mL, or about 50 ng / mL.

[0082] In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, increases serum creatine kinase (CK) levels to 10 U / L or greater, 20 U / L or greater, 30 U / L or greater, 40 U / L or greater, 50 U / L or greater, 60 U / L or greater, 70 U / L or greater, 80 U / L or greater, 90 U / L or greater, 100 U / L or greater, 200 U / L or greater, 300 U / L or greater, or or greater, 400 U / L or greater, 500 U / L or greater, 600 U / L or greater, 700 U / L or greater, 800 U / L or greater, 900 U / L or greater, 1000 U / L or greater, 1100 U / L or greater, 1200 U / L or greater, 1300 U / L or greater, 1400 U / L or greater, or 1500 U / L or greater. In some embodiments, CK levels are reduced by about 500 U / L to 1500 U / L, about 800 U / L to 1400 U / L, about 1000 U / L to 1300 U / L, or about 1100 U / L to 1300 U / L.

[0083] In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's serum creatine kinase (CK) level to between about 400 U / L and about 1,500 U / L. In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered to a subject to increase serum creatine kinase (CK) levels by about 400 U / L to about 500 U / L, about 400 U / L to about 600 U / L, about 400 U / L to about 700 U / L, about 400 U / L to about 800 U / L, about 400 U / L to about 900 U / L, about 400 U / L to about 1,000 U / L, about 400 U / L to about 1,100 U / L, about 400 U / L to about 1,200 U / L, about 400 U / L to about 1,300 U / L, or about 400 U / L. L ~ approx. 1,400U / L, approx. 400U / L ~ approx. 1,500U / L, approx. 500U / L ~ approx. 600U / L, approx. 500U / L ~ approx. 700U / L, approx. 500U / L ~ approx. 800U / L, approx. 500U / L ~ approx. 900U / L, approx. 500U / L ~ approx. 1,000U / L , about 500U / L to about 1,100U / L, about 500U / L to about 1,200U / L, about 500U / L to about 1,300U / L, about 500U / L to about 1,400U / L, about 500U / L to about 1,500U / L, about 600U / L to about 700U / L, about 600U / L ~ approx. 800U / L, approx. 600U / L ~ approx. 900U / L, approx. 600U / L ~ approx. 1,000U / L, approx. 600U / L ~ approx. 1,100U / L, approx. 600U / L ~ approx. 1,200U / L, approx. 600U / L ~ approx. 1,300U / L, approx. 00U / L, about 600U / L to about 1,500U / L, about 700U / L to about 800U / L, about 700U / L to about 900U / L, about 700U / L to about 1,000U / L, about 700U / L to about 1,100U / L, about 700U / L to about 1,200U / L, about 7 00U / L~Approx. 1,300U / L, Approx. 700U / L~Approx. 1,400U / L, Approx. 700U / L~Approx. 1,500U / L, Approx. 800U / L~Approx. 900U / L, Approx. 800U / L~Approx. 1,000U / L, Approx. 800U / L~Approx. 1,100U / L, Approx. 800U / L ~1,200U / L, 800U / L~1,300U / L, 800U / L~1,400U / L, 800U / L~1,500U / L, 900U / L~1,000U / L, 900U / L~1,100U / L, 900U / L~1,200U / L, approximately 900U / L to approximately 1,300U / L, approximately 900U / L to approximately 1,400U / L, approximately 900U / L to approximately 1,500U / L, approximately 1,000U / L to approximately 1,100U / L, approximately 1,000U / L to approximately 1,20 0U / L, approximately 1,000U / L to approximately 1,300U / L, approximately 1,000U / L to approximately 1,400U / L, approximately 1,000U / L to approximately 1,500U / L, approximately 1,100U / L to approximately 1,200U / L, approximately 1,100U / L to approximately 1 It is administered in an amount sufficient to maintain or reduce the blood cholesterol level to about 1,300 U / L, about 1,100 U / L to about 1,400 U / L, about 1,100 U / L to about 1,500 U / L, about 1,200 U / L to about 1,300 U / L, about 1,200 U / L to about 1,400 U / L, about 1,200 U / L to about 1,500 U / L, about 1,300 U / L to about 1,400 U / L, about 1,300 U / L to about 1,500 U / L, or about 1,400 U / L to about 1,500 U / L. In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's serum creatine kinase (CK) level to about 400 U / L, about 500 U / L, about 600 U / L, about 700 U / L, about 800 U / L, about 900 U / L, about 1,000 U / L, about 1,100 U / L, about 1,200 U / L, about 1,300 U / L, about 1,400 U / L, or about 1,500 U / L. In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's serum creatine kinase (CK) level to at least about 400 U / L, about 500 U / L, about 600 U / L, about 700 U / L, about 800 U / L, about 900 U / L, about 1,000 U / L, about 1,100 U / L, about 1,200 U / L, about 1,300 U / L, or about 1,400 U / L. In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce a subject's serum creatine kinase (CK) level to a maximum of about 500 U / L, about 600 U / L, about 700 U / L, about 800 U / L, about 900 U / L, about 1,000 U / L, about 1,100 U / L, about 1,200 U / L, about 1,300 U / L, about 1,400 U / L, or about 1,500 U / L.

[0084] In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, is administered to a subject in an amount sufficient to reduce myoglobin levels by 10 ng / mL or more, 20 ng / mL or more, 30 ng / mL or more, 40 ng / mL or more, 50 ng / mL or more, 60 ng / mL or more, 70 ng / mL or more, 80 ng / mL or more, 90 ng / mL or more, 100 ng / mL or more, 200 ng / mL or more, 300 ng / mL or more, 400 ng / mL or more, 500 ng / mL or more, 600 ng / mL or more, 700 ng / mL or more, 800 ng / mL or more, 900 ng / mL or more, or 1000 ng / mL or more.

[0085] In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's myoglobin level to about 100 U / L to about 400 U / L. In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's myoglobin level to about 100 U / L to about 150 U / L, about 100 U / L to about 200 U / L, about 100 U / L to about 250 U / L, about 100 U / L to about 300 U / L, about 100 U / L to about 350 U / L, about 100 U / L to about 400 U / L, about 150 U / L to about 200 U / L, about 150 U / L to about 250 U / L, about 150 U / L to about 300 U / L, about 150 U / L to about 150 U / L. It is administered in an amount sufficient to maintain or reduce the blood cholesterol level to about 350 U / L, about 150 U / L to about 400 U / L, about 200 U / L to about 250 U / L, about 200 U / L to about 300 U / L, about 200 U / L to about 350 U / L, about 200 U / L to about 400 U / L, about 250 U / L to about 300 U / L, about 250 U / L to about 350 U / L, about 250 U / L to about 400 U / L, about 300 U / L to about 350 U / L, about 300 U / L to about 400 U / L, or about 350 U / L to about 400 U / L. In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's myoglobin level to about 100 U / L, about 150 U / L, about 200 U / L, about 250 U / L, about 300 U / L, about 350 U / L, or about 400 U / L. In certain embodiments, the fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the subject's myoglobin level to at least about 100 U / L, about 150 U / L, about 200 U / L, about 250 U / L, about 300 U / L, or about 350 U / L. In certain embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce a subject's myoglobin level to a maximum of about 150 U / L, about 200 U / L, about 250 U / L, about 300 U / L, about 350 U / L, or about 400 U / L.

[0086] In some embodiments, a fast-twitch myosin (type II) inhibitor, e.g., Compound 1, is administered to a subject in an amount sufficient to increase serum creatinine levels by 1 mg / dL or more, 0.8 mg / dL or more, 0.6 mg / dL or more, 0.5 mg / dL or more, 0.4 mg / dL or more, 0.3 mg / dL or more, 0.2 mg / dL or more, or 0.1 mg / dL or more.

[0087] Administration of Compound 1 or a salt thereof can reduce involuntary muscle contractions. The involuntary muscle contractions can be reduced by 20% to 90% compared to the involuntary muscle contractions before administration of the inhibitor. In some cases, the involuntary muscle contractions can be reduced by at least about 20% compared to the involuntary muscle contractions before treatment. In some cases, the involuntary muscle contractions can be reduced by up to about 90% compared to the involuntary muscle contractions before treatment. In some cases, the involuntary muscle contractions are reduced by about 20% to about 25%, about 20% to about 30%, about 20% to about 40%, about 20% to about 50%, about 20% to about 70%, about 20% to about 75%, about 20% to about 80%, about 20% to about 85%, about 20% to about 90%, about 25% to about 30%, about 25% to about 40%, about 25% to about 50%, about 25% to about 70%, about 25% to about 75%, about 25% to about 80%, about 25% to about 85%, about 25% to about 90%, about 30% to about 40%, about 30% to about 50%, about 30% to about 70%, about 30% to about 75%, about 30% The concentration may be reduced by about 80%, about 30% to about 85%, about 30% to about 90%, about 40% to about 50%, about 40% to about 70%, about 40% to about 75%, about 40% to about 80%, about 40% to about 85%, about 40% to about 90%, about 50% to about 70%, about 50% to about 75%, about 50% to about 80%, about 50% to about 85%, about 50% to about 90%, about 70% to about 75%, about 70% to about 80%, about 70% to about 85%, about 70% to about 90%, about 75% to about 80%, about 75% to about 85%, about 75% to about 90%, about 80% to about 85%, about 80% to about 90%, or about 85% to about 90%. In some cases, involuntary muscle contractions may be reduced by about 20%, about 25%, about 30%, about 40%, about 50%, about 70%, about 75%, about 80%, about 85%, or about 90% compared to involuntary muscle contractions before treatment.

[0088] Compound 1 or its salt can be used to improve the activities of daily living (ADL) or habitual physical activity in subjects, because mature, functionally intact muscles can be restored.Examples of ADL or habitual activity include, but are not limited to, stair climbing, time to stand up, timed chair rise, habitual walking speed, North Star Ambulation Assessment (NSAA), incremental / endurance shuttle walk, and 6-minute walk distance test.The level or capacity of ADL or habitual physical activity can be measured before and after administration of skeletal muscle inhibitors.Inhibition of skeletal muscle contraction may not affect ADL or habitual physical activity.In some cases, inhibition of skeletal muscle contraction may not appreciably affect ADL or habitual physical activity. When used herein with respect to ADL or habitual physical activity, the phrase refers to a level of ADL or habitual activity that is appreciably reduced by less than 20%, less than 15%, less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1%, less than 0.5%, or even less than 0.1% compared to the ADL or habitual activity prior to administration of the inhibitor.

[0089] In some embodiments, administration of Compound 1 or a salt thereof affects a subject's ADL or habitual physical activity. In some embodiments, the North Star Ambulatory Assessment is used to assess the progression of neuromuscular disease in a subject over time. For example, FIG. 1 shows the NSAA scores of human subjects diagnosed with Becker muscular dystrophy over time. On average, there is an estimated mean annual decrease of -1.22 for patients aged 10 to 32 years. In some embodiments, administration of Compound 1 or a salt thereof slows the rate at which neuromuscular disease progresses, as determined by the NSAA score. In some embodiments, administration of Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -1.22. In some embodiments, administration of Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -1. In some embodiments, administration of Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.5. In some embodiments, administration of Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.4. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than -0.3. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than -0.2. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than -0.1. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0.1. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0.2. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0.3. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0.4. In some embodiments, administration of Compound 1 or a salt increases the mean annual NSAA score to greater than 0.5. In some embodiments, administration of Compound 1 or a salt increases the mean annualized NSAA score to greater than 1.0. In some embodiments, administration of Compound 1 or a salt increases the mean annualized NSAA score to greater than 2.0.In some embodiments, administration of Compound 1 or a salt increases the average annual NSAA score to greater than 3.0. In some embodiments, administration of Compound 1 or a salt increases the average annual NSAA score to greater than 4.0. In some embodiments, administration of Compound 1 or a salt increases the average annual NSAA score to greater than 5.0. In some embodiments, administration of Compound 1 or a salt increases the average annual NSAA score to greater than 10.0. In some embodiments, administration of Compound 1 or a salt increases the average monthly NSAA score to at least -0.1, 0, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 2.0, 3.0, 4.0, 5.0, or 10.0.

[0090] In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than about 10. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than 2. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than 1.5. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than 1. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than 0.5. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than 0. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1.2 to greater than -0.5. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1 to greater than 2. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -0.5 to greater than 2. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about 0 to greater than 5. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -1 to greater than 5. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about -0.5 to greater than 4. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about 0 to greater than 3. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score from about 2 to greater than 10. In some embodiments, administration of a fast-twitch myosin (type II) inhibitor increases the patient's NSAA score by about -1.2, about -1, about -0.75, about -0.5, about -0.25, about 0, about 0.25, about 0.5, about 0.75, about 1, about 1.5, or about 2, about 3, about 4, about 5, about 6, about 7, about 8, or greater than about 10.

[0091] In some embodiments, the patient's NSAA score increases after administration of a muscle myosin inhibitor, e.g., Compound 1. In some embodiments, the fast myosin (type II) inhibitor is administered for about 1 month to about 24 months. In some embodiments, the patient's NSAA score increases after administration of a muscle myosin inhibitor, e.g., Compound 1. In some embodiments, the fast myosin (type II) inhibitor is administered for about 2 months to about 12 months. In some embodiments, the patient's NSAA score increases after administration of a muscle myosin inhibitor, e.g., Compound 1. In some embodiments, the fast myosin (type II) inhibitor is administered for about 4 months to about 12 months. In some embodiments, the patient's NSAA score increases after administration of a muscle myosin inhibitor, e.g., Compound 1. In some embodiments, the fast myosin (type II) inhibitor is administered for about 6 months to about 18 months. In some embodiments, the patient's NSAA score increases after administration of a muscle myosin inhibitor, e.g., Compound 1. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for about 2 months to about 6 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 10 months, about 12 months, about 18 months, or about 24 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for at least about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 10 months, about 12 months, or about 18 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for up to about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 10 months, about 12 months, about 18 months, or about 24 months.

[0092] Administer a fast-twitch myosin (type II) inhibitor, such as Compound 1, to the subject in an amount sufficient to maintain or improve one or more functional measures of the subject. In some embodiments, the functional measure is selected from the following: 100-meter timed test speed compared to the 100-meter timed test speed before treatment; 4-step stair climbing timed speed compared to the 4-step stair climbing timed speed before treatment; 10-meter walking / running speed compared to the 10-meter walking / running speed before treatment; maximal elbow flexion strength compared to the maximal elbow flexion strength before treatment; maximal knee extension strength compared to the maximal knee extension strength before treatment; and maximal grip strength compared to the maximal grip strength value before treatment. In some embodiments, the functional measure comprises the assessment of the patient's ability to perform Gowers sign, walk, stand from a chair, stand on one leg, climb a box with a step, descend a box with a step, sit, stand from the floor, lift head, stand on heels, jump on two legs, hop on one leg, or run.

[0093] In some embodiments, the fast-twitch myosin (type II) inhibitor is administered in an amount sufficient to maintain one or more functional measures of the subject compared to pre-treatment values. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered in an amount sufficient to improve one or more functional measures of the subject compared to pre-treatment values. In some embodiments, the one or more functional measures are maintained within 20%, 30%, 40%, or 50% of pre-treatment values. In some embodiments, the one or more functional measures are maintained for 2 months or more, 4 months or more, 6 months or more, 8 months or more, 12 months or more, 18 months or more, 24 months or more, or 36 months or more.

[0094] Skeletal muscle contraction or force in a subject can be measured before and after administration of Compound 1 or a salt thereof. Such measurements can be performed to generate a dose-response curve for Compound 1 or a salt thereof. The dosage of Compound 1 or a salt thereof can be adjusted by about 5% to 50% compared to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dosage of the skeletal muscle contraction inhibitor can be adjusted by at least about 5% compared to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dosage of the skeletal muscle contraction inhibitor can be adjusted by up to about 50% compared to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dose of the skeletal muscle contraction inhibitor may be about 5% to about 10%, about 5% to about 15%, about 5% to about 20%, about 5% to about 25%, about 5% to about 30%, about 5% to about 35%, about 5% to about 40%, about 5% to about 50%, about 10% to about 15%, about 10% to about 20%, about 10% to about 25%, about 10% to about 30%, about 10% to about 35%, about 10% to about 40%, about 10% to about 50%, about 15% to about 20%, about 15% to about It can be adjusted to 25%, about 15% to about 30%, about 15% to about 35%, about 15% to about 40%, about 15% to about 50%, about 20% to about 25%, about 20% to about 30%, about 20% to about 35%, about 20% to about 40%, about 20% to about 50%, about 25% to about 30%, about 25% to about 35%, about 25% to about 40%, about 25% to about 50%, about 30% to about 35%, about 30% to about 40%, about 30% to about 50%, about 35% to about 40%, about 35% to about 50%, or about 40% to about 50%. In some cases, the dosage of the skeletal muscle contraction inhibitor can be adjusted by about 10%, about 12%, about 15%, about 18%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, or about 50% compared to the dose that reduces type II skeletal muscle contraction by 90%. Skeletal muscle contraction can be measured before and after administration of the skeletal muscle contraction inhibitor by muscle strength testing after nerve stimulation using surface electrodes (e.g., plantar flexion after peroneal nerve stimulation in the leg), single limb assay, heart rate monitor, or activity monitor, or equivalent.

[0095] The tidal volume of the lungs of a subject can be measured before and after administration of Compound 1 or its salt.Administration may not inhibit the tidal volume of the lungs.In some cases, administration may not appreciably inhibit the tidal volume of the lungs.In certain embodiments relating to tidal lung volume of the lungs, this phrase refers to the tidal volume of the lungs being reduced by less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1%, less than 0.5%, or less than 0.1% compared to the tidal volume of the lungs before administration of the inhibitor.The tidal volume of the lungs of a subject can be measured using forced volume in one second test (FEV1) or forced vital capacity test (FVC), or its equivalent test.

[0096] The smooth muscle contraction in subject can be measured before and after administration of skeletal muscle contraction inhibitor.Inhibition of skeletal muscle contraction may not inhibit smooth muscle contraction.In some cases, inhibition of skeletal muscle contraction may not inhibit smooth muscle contraction to an appreciable extent.When used herein in relation to smooth muscle contraction, this phrase refers to smooth muscle contraction that is not appreciably reduced by less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1%, less than 0.5%, or even less than 0.1% compared to the smooth muscle contraction before administration of inhibitor.The smooth muscle contraction in subject can be evaluated by measuring the blood pressure of subject.

[0097] Neuromuscular coupling in a subject can be measured before and after administration of Compound 1 or its salt. The inhibition of skeletal muscle contraction by the inhibitors described herein may not impair the nerve conduction, neurotransmitter release, or electrical depolarization of skeletal muscle in a subject. In some cases, the inhibition of skeletal muscle contraction may not appreciably impair the neuromuscular coupling in a subject. When used herein with respect to neuromuscular coupling, this phrase refers to the level of neuromuscular coupling in a subject that is appreciably reduced by less than 10%, less than 8%, less than 6%, less than 4%, less than 2%, less than 1%, less than 0.5%, or less than 0.1% compared to the level of neuromuscular coupling in a subject before administration of the inhibitor. Neuromuscular coupling in a subject can be evaluated by measuring the nerve-induced electrical depolarization of skeletal muscle by recording the electrical activity generated by skeletal muscle after electrical stimulation or voluntary stimulation using electromyography (EMG) with surface or needle electrodes.

[0098] The pain may be associated with a neuromuscular condition disclosed herein. Accordingly, disclosed herein is a method of reducing pain associated with a neuromuscular condition, comprising administering a fast-twitch myosin (type II) inhibitor to a subject in need thereof. In some embodiments, the pain is diffuse. In some embodiments, the pain is in the spine of the subject. In some embodiments, the pain is in the calf of the subject. In some embodiments, the pain is associated with Becker muscular dystrophy (BMD), Duchenne muscular dystrophy (DMD), limb-girdle muscular dystrophy (LGMD), McArdle's disease, and other neuromuscular conditions.

[0099] In some embodiments, a fast myosin (type II) inhibitor is administered to a subject exhibiting a dystrophin gene mutation selected from del 45-48, del 45-47, del 48, del x-51, del 45-55, and del 48-49.

[0100] In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma PK level of 500 ng / mL or higher, 1,000 ng / mL or higher, 1,200 ng / mL or higher, 1,400 ng / mL or higher, 1,800 ng / mL or higher, 2,000 ng / mL or higher, 2,200 ng / mL or higher, 2,300 ng / mL or higher, 2,400 ng / mL or higher, 2,500 ng / mL or higher, 2,800 ng / mL or higher, 3,000 ng / mL or higher, or 4,000 ng / mL or higher. In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma PK value of about 500 ng / mL to 4,000 ng / mL, about 500 ng / mL to 4,000 ng / mL, about 1,000 ng / mL to 3,000 ng / mL, about 1,300 ng / mL to 2,800 ng / mL, about 1,400 ng / mL to 2,600 ng / mL, about 2,000 ng / mL to 2,600 ng / mL, or about 2,200 ng / mL to 2,400 ng / mL.

[0101] In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of about 10 ng / mL to about 200 ng / mL. In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of about 10 ng / mL to about 20 ng / mL, about 10 ng / mL to about 30 ng / mL, about 10 ng / mL to about 40 ng / mL, about 10 ng / mL to about 50 ng / mL, about 10 ng / mL to about 60 ng / mL, about 10 ng / mL to about 70 ng / mL, about 10 ng / mL to about 80 ng / mL, about 10 ng / mL to about 90 ng / mL, about 10 ng / mL to about 100 ng / mL, about 10 ng / mL to about 150 ng / mL, about 10 ng / mL to about 200 ng / mL, or about 20 ng / mL to about 30 ng / mL. L, about 20ng / mL to about 40ng / mL, about 20ng / mL to about 50ng / mL, about 20ng / mL to about 60ng / mL, about 20ng / mL to about 70ng / mL, about 20ng / mL to about 80ng / mL, about 20ng / mL to about 90ng / mL, about 20ng / mL mL~about 100ng / mL, about 20ng / mL~about 150ng / mL, about 20ng / mL~about 200ng / mL, about 30ng / mL~about 40ng / mL, about 30ng / mL~about 50ng / mL, about 30ng / mL~about 60ng / mL, about 30ng / mL~about 70 ng / mL, about 30ng / mL to about 80ng / mL, about 30ng / mL to about 90ng / mL, about 30ng / mL to about 100ng / mL, about 30ng / mL to about 150ng / mL, about 30ng / mL to about 200ng / mL, about 40ng / mL to about 50ng / mL , about 40ng / mL to about 60ng / mL, about 40ng / mL to about 70ng / mL, about 40ng / mL to about 80ng / mL, about 40ng / mL to about 90ng / mL, about 40ng / mL to about 100ng / mL, about 40ng / mL to about 150ng / mL, about 40ng / mL mL~about 200ng / mL, about 50ng / mL~about 60ng / mL, about 50ng / mL~about 70ng / mL, about 50ng / mL~about 80ng / mL, about 50ng / mL~about 90ng / mL, about 50ng / mL~about 100ng / mL, about 50ng / mL~about 150 ng / mL, approximately 50ng / mL to approximately 200ng / mL, approximately 60ng / mL to approximately 70ng / mL, approximately 60ng / mL to approximately 80ng / mL, approximately 60ng / mL to approximately 90ng / mL, approximately 60ng / mL to approximately 100ng / mL, approximately 60ng / mL to approximately 150ng / mL,Approximately 60ng / mL to approximately 200ng / mL, approximately 70ng / mL to approximately 80ng / mL, approximately 70ng / mL to approximately 90ng / mL, approximately 70ng / mL to approximately 100ng / mL, approximately 70ng / mL to Approximately 150ng / mL, approximately 70ng / mL to approximately 200ng / mL, approximately 80ng / mL to approximately 90ng / mL, approximately 80ng / mL to approximately 100ng / mL, approximately 80ng / mL to approximately 150ng / m In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of about 10 ng / mL, about 20 ng / mL, about 30 ng / mL, about 40 ng / mL, about 50 ng / mL, about 60 ng / mL, about 70 ng / mL, about 80 ng / mL, about 90 ng / mL, about 100 ng / mL, about 150 ng / mL, about 90 ng / mL, about 200 ng / mL, about 100 ng / mL, about 150 ng / mL, or about 200 ng / mL. In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of about 10 ng / mL, about 20 ng / mL, about 30 ng / mL, about 40 ng / mL, about 50 ng / mL, about 60 ng / mL, about 70 ng / mL, about 80 ng / mL, about 90 ng / mL, about 100 ng / mL, about 150 ng / mL, or about 200 ng / mL. In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of at least about 10 ng / mL, about 20 ng / mL, about 30 ng / mL, about 40 ng / mL, about 50 ng / mL, about 60 ng / mL, about 70 ng / mL, about 80 ng / mL, about 90 ng / mL, about 100 ng / mL, or about 150 ng / mL. In some embodiments, Compound 1 is administered to a subject in an amount sufficient to maintain a plasma concentration of at most about 20 ng / mL, about 30 ng / mL, about 40 ng / mL, about 50 ng / mL, about 60 ng / mL, about 70 ng / mL, about 80 ng / mL, about 90 ng / mL, about 100 ng / mL, about 150 ng / mL, or about 200 ng / mL.

[0102] The fast-twitch myosin (type II) inhibitor can be administered for various periods. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered for more than 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, or more than 12 months.

[0103] In some embodiments, Compound 1 is administered. In some embodiments, Compound 1 is administered at a daily dose of about 1 mg to about 10 mg. In some embodiments, Compound 1 is administered at a daily dose of about 1 mg to about 2 mg, about 1 mg to about 2.5 mg, about 1 mg to about 3 mg, about 1 mg to about 4 mg, about 1 mg to about 5 mg, about 1 mg to about 6 mg, about 1 mg to about 7 mg, about 1 mg to about 7.5 mg, about 1 mg to about 8 mg, about 1 mg to about 9 mg, about 1 mg to about 10 mg, about 2 mg to about 2.5 mg, about 2 mg to about 3 mg, about 2 mg to about 4 mg, about 2 mg to about 5 mg, about 2 mg to about 6 mg, about 2 mg to about 6 mg, about 2 mg to about 7 mg, about 2 mg to about 8 mg, about 2 mg to about 9 mg, about 1 mg to about 10 mg, about 2 mg to about 2.5 mg, about 2 mg to about 3 mg, about 2 mg to about 4 mg, about 2 mg to about 5 mg, about 2 mg to about 6 ... g ~ about 7mg, about 2mg - about 7.5mg, about 2mg - about 8mg, about 2mg - about 9mg, about 2mg - about 10mg, about 2.5mg - about 3mg, about 2.5mg - about 4mg, about 2.5mg - about 5mg, about 2.5mg Approximately 6 mg, approximately 2.5 mg to approximately 7 mg, approximately 2.5 mg to approximately 7.5 mg, approximately 2.5 mg to approximately 8 mg, approximately 2.5 mg to approximately 9 mg, approximately 2.5 mg to approximately 10 mg, approximately 3 mg to approximately 4 mg, approximately 3 mg to approximately 5 mg, approximately 3 mg Approximately 6 mg, approximately 3 mg to approximately 7 mg, approximately 3 mg to approximately 7.5 mg, approximately 3 mg to approximately 8 mg, approximately 3 mg to approximately 9 mg, approximately 3 mg to approximately 10 mg, approximately 4 mg to approximately 5 mg, approximately 4 mg to approximately 6 mg, approximately 4 mg to approximately 7 mg, approximately 4 mg Approximately 7.5 mg, approximately 4 mg to approximately 8 mg, approximately 4 mg to approximately 9 mg, approximately 4 mg to approximately 10 mg, approximately 5 mg to approximately 6 mg, approximately 5 mg to approximately 7 mg, approximately 5 mg to approximately 7.5 mg, approximately 5 mg to approximately 8 mg, approximately 5 mg to approximately 9 mg, approximately 5 m In some embodiments, Compound 1 is administered at a daily dose of about 1 mg, about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 7.5 mg, about 8 mg, about 9 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 6 mg, about 7 mg, about 7.5 mg, about 8 mg, about 7 mg, about 9 mg, about 7 mg, about 10 mg, about 7.5 mg, about 8 mg, about 7.5 mg, about 9 mg, about 7.5 mg, about 10 mg, about 8 mg, about 9 mg, about 8 mg, about 10 mg, or about 9 mg to about 10 mg. In some embodiments, Compound 1 is administered at a daily dose of about 1 mg, about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 7.5 mg, about 8 mg, about 9 mg, or about 10 mg. In some embodiments, Compound 1 is administered at a daily dose of at least about 1 mg, about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 7.5 mg, about 8 mg, or about 9 mg.In some embodiments, Compound 1 is administered at a daily dose of up to about 2 mg, about 2.5 mg, about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 7.5 mg, about 8 mg, about 9 mg, or about 10 mg. In some embodiments, Compound 1 is administered at a daily dose of about 2.5 mg to about 20 mg. In some embodiments, Compound 1 is from about 2.5 mg to about 5 mg, from about 2.5 mg to about 7.5 mg, from about 2.5 mg to about 10 mg, from about 2.5 mg to about 12.5 mg, from about 2.5 mg to about 15 mg, from about 2.5 mg to about 17.5 mg, from about 2.5 mg to about 20 mg, from about 5 mg to about 7.5 mg, from about 5 mg to about 10 mg, from about 5 mg to about 12.5 mg, from about 5 mg to about 15 mg, from about 5 mg to about 17.5 mg, from about 5 mg to about 20 mg, from about 7.5 mg to about 10 mg, or from about 7.5 mg to about 10 mg. In some embodiments, Compound 1 is administered at a daily dose of about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 10 mg, about 12.5 mg, about 10 mg, about 15 mg, about 10 mg, about 17.5 mg, about 10 mg, about 20 mg, about 12.5 mg, about 15 mg, about 12.5 mg, about 17.5 mg, about 15 mg, about 20 mg, or about 17.5 mg to about 20 mg. In some embodiments, Compound 1 is administered at a daily dose of about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg. In some embodiments, Compound 1 is administered at a daily dose of at least about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, or about 17.5 mg. In some embodiments, Compound 1 is administered at a daily dose of up to about 5 mg, about 7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg.

[0104] In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at a constant dosage throughout the entire treatment. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at a constant dosage throughout approximately 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at a constant dosage for more than 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, or more than 12 months. In some embodiments, the constant dosage is greater than 1 mg / day, more than 2.5 mg / day, more than 5 mg / day, more than 10 mg / day, more than 15 mg / day, more than 20 mg / day, more than 25 mg / day, more than 30 mg / day, more than 35 mg / day, or more than 40 mg / day. In some embodiments, the constant dosage is less than 1 mg / day, less than 2.5 mg / day, less than 5 mg / day, less than 10 mg / day, less than 15 mg / day, less than 20 mg / day, less than 25 mg / day, less than 30 mg / day, less than 35 mg / day, less than 40 mg / day, less than 45 mg / day, less than 50 mg / day, less than 55 mg / day, or less than 60 mg / day. In some embodiments, the constant dosage is about 1 mg / day, 2 mg / day, 2.5 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 7.5 mg / day, 8 mg / day, 9 mg / day, 10 mg / day, 11 mg / day, 12 mg / day, 12.5 mg / day, 13 mg / day, 14 mg / day, 15 mg / day, 16 mg / day, 17 mg / day, 17.5 mg / day, 18 mg / day, 19 mg / day, or 20 mg / day. In some preferred embodiments, the constant dosage is about 10 mg / day.

[0105] In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying dosages throughout the entire treatment. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying dosages throughout about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying dosages for more than 1 month, more than 2 months, more than 3 months, more than 4 months, more than 5 months, more than 6 months, more than 7 months, more than 8 months, more than 9 months, more than 10 months, more than 11 months, or more than 12 months. In a specific embodiment, the fast-twitch myosin (type II) inhibitor is administered at gradually decreasing dosages throughout the treatment. In a specific embodiment, the fast-twitch myosin (type II) inhibitor is administered at gradually increasing dosages throughout the treatment. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at x mg / day, followed by (x + 5) mg / day, followed by (x + 5 x 2) mg / day, followed by (x + 5 x 3) mg / day, followed by (x + 5 x 4) mg / day, and finally followed by (x + 5 x y) mg / day, where x can be a number between 1 and 100, 5 and 100, 5 and 50, 5 and 20, or 5 and 10, and y can be a number between 1 and 10. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at x mg / day, followed by (x + 5) mg / day, and further followed by (x + 10) mg / day, where x can be between 1 and 100, 5 and 100, 5 and 50, 5 and 20, or 5 and 10. In some embodiments, the fast myosin (type II) inhibitor is administered at 10 mg / day, followed by 15 mg / day, and further followed by 20 mg / day.

[0106] In some embodiments, where the fast-twitch myosin (type II) inhibitor is administered at varying dosages throughout treatment, the varying dosages last for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some specific embodiments, the fast-twitch myosin (type II) inhibitor is administered at 10 mg / day in months 1 and 2, followed by 15 mg / day in months 3-6, and further followed by 20 mg / day in months 7-12.

[0107] The fast-twitch myosin (type II) inhibitor may be administered at varying dosages, which are determined in real time based on one or more biomarkers and / or plasma concentrations of the fast-twitch myosin (type II) inhibitor in the subject. In some embodiments, the varying dosages are determined based on whether they are sufficient to reduce one or more biomarkers (e.g., creatine kinase, lactate dehydrogenase, myoglobin, TNNI2). In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying dosages to maintain the plasma concentration of the fast-twitch myosin (type II) inhibitor in the subject within an optimal range. In some embodiments, the fast myosin (type II) inhibitor is between 5 ng / ml and 200 ng / ml, between 10 ng / ml and 200 ng / ml, between 10 ng / ml and 150 ng / ml, between 10 ng / ml and 140 ng / ml, between 10 ng / ml and 130 ng / ml, between 10 ng / ml and 120 ng / ml, between 10 ng / ml and 110 ng / ml, between 10 ng / ml and 100 ng / ml , between 10ng / ml and 80ng / ml, between 20ng / ml and 70ng / ml, between 40ng / ml and 60ng / ml, between 40ng / ml and 100ng / ml, between 20ng / ml and 200ng / ml, between 20ng / ml and 150ng / ml, between 20ng / ml and 140ng / ml, between 20ng / ml and 130ng / ml, between 20ng / ml and 120ng / ml, 20ng / ml and Between 110ng / ml, between 20ng / ml and 100ng / ml, between 30ng / ml and 200ng / ml, between 30ng / ml and 150ng / ml, between 30ng / ml and 140ng / ml, between 30ng / ml and 130ng / ml, between 30ng / ml and 120ng / ml, between 30ng / ml and 110ng / ml, between 30ng / ml and 100ng / ml, between 40ng / ml and 200ng The fast-twitch myosin (type II) inhibitor is administered at various dosages to maintain a plasma concentration in the subject of between 40 ng / ml and 150 ng / ml, between 40 ng / ml and 140 ng / ml, between 40 ng / ml and 130 ng / ml, between 40 ng / ml and 120 ng / ml, between 40 ng / ml and 110 ng / ml, or between 40 ng / ml and 100 ng / ml.In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying doses to maintain a plasma concentration of the fast-twitch myosin (type II) inhibitor in the subject between 10 ng / ml and 100 ng / ml, between 10 ng / ml and 100 ng / ml, between 10 ng / ml and 100 ng / ml, between 20 ng / ml and 100 ng / ml, or between 30 ng / ml and 100 ng / ml. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at varying doses to maintain a plasma concentration of the fast-twitch myosin (type II) inhibitor in the subject greater than 10 ng / ml, greater than 15 ng / ml, greater than 20 ng / ml, greater than 25 ng / ml, greater than 30 ng / ml, greater than 35 ng / ml, or greater than 40 ng / ml. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at various doses to maintain a plasma concentration of the fast-twitch myosin (type II) inhibitor in the subject of less than 70 ng / ml, less than 60 ng / ml, less than 50 ng / ml, less than 40 ng / ml, or less than 30 ng / ml. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at a dose of between 15 ng / ml and 70 ng / ml, between 15 ng / ml and 60 ng / ml, between 15 ng / ml and 50 ng / ml, between 15 ng / ml and 40 ng / ml, between 15 ng / ml and 30 ng / ml, between 15 ng / ml and 20 ng / ml, between 20 ng / ml and 70 ng / ml, between 25 ng / ml and 70 ng / ml, between 30 ng / ml and 70 ng / ml. and / or at various dosages to maintain a plasma concentration of the fast twitch myosin (type II) inhibitor in the subject between 35 ng / ml and 70 ng / ml, between 40 ng / ml and 70 ng / ml, between 45 ng / ml and 70 ng / ml, between 50 ng / ml and 70 ng / ml, between 55 ng / ml and 70 ng / ml, between 60 ng / ml and 70 ng / ml, or between 65 ng / ml and 70 ng / ml.

[0108] In some embodiments, fast myosin (type II) inhibitor is abundant in muscle.Therefore, in some embodiments, fast myosin (type II) inhibitor is administered at various dosages to maintain the muscle concentration of the fast myosin (type II) inhibitor in the subject to be greater than 500ng / g, greater than 600ng / g, greater than 700ng / g, greater than 800ng / g, greater than 900ng / g, greater than 1000ng / g, greater than 1100ng / g, greater than 1200ng / g, greater than 1300ng / g, greater than 1400ng / g, greater than 1500ng / g, greater than 2000ng / g or greater than 3000ng / g. In some embodiments, a fast-twitch myosin (type II) inhibitor is administered at various dosages to maintain a muscle concentration of said fast-twitch myosin (type II) inhibitor in the subject of less than 10,000 ng / g, less than 9,000 ng / g, less than 8,000 ng / g, less than 7,000 ng / g, less than 6,000 ng / g, less than 5,000 ng / g, less than 4,000 ng / g, or less than 3,000 ng / g. In some embodiments, a fast-twitch myosin (type II) inhibitor is administered at varying dosages to maintain a muscle concentration of the fast-twitch myosin (type II) inhibitor in the subject between 1000 ng / g and 4000 ng / g, between 1000 ng / g and 3000 ng / g, between 1000 ng / g and 2000 ng / g, between 2000 ng / g and 4000 ng / g, between 2000 ng / g and 4000 ng / g, or between 3000 ng / g and 4000 ng / g.

[0109] The compositions and methods described herein may be useful as pharmaceutical compositions for administration to subjects in need thereof.The pharmaceutical composition may include a fast myosin (type II) inhibitor and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents, and / or thickeners.In some embodiments, the fast myosin (type II) inhibitor is a compound or salt thereof described in WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615.In some embodiments, the fast myosin (type II) inhibitor is described in WO2020097266. In some embodiments, the muscle myosin inhibitor is a compound of formula (I), also referred to as Compound 1: [ka] is.

[0110] Pharmaceutical compositions containing fast myosin (type II) inhibitors can be formulated using one or more physiologically acceptable carriers, including excipients and auxiliary agents.The formulation can be modified according to the selected route of administration.Pharmaceutical compositions containing compounds, salts, or conjugates can be produced, for example, by lyophilizing, mixing, dissolving, emulsifying, encapsulating, or enclosing the compounds, salts, or conjugates.Pharmaceutical compositions can also contain compounds, salts, or conjugates in free base form or pharmaceutically acceptable salt form.

[0111] Pharmaceutical compositions containing fast myosin (type II) inhibitors may include formulating either the compound, salt, or conjugate with one or more inert pharmaceutically acceptable excipients or carriers to form solid, semi-solid, or liquid compositions.Solid compositions may include, for example, powders, tablets, dispersible granules, and capsules, and in some embodiments, the solid compositions further contain non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and other pharmaceutically acceptable additives.Alternatively, the compound, salt, or conjugate may be lyophilized or in powder form for reconstitution with a suitable vehicle, for example, sterile pyrogen-free water, before use.

[0112] Pharmaceutical compositions containing fast-twitch myosin (type II) inhibitors may contain at least one active ingredient (e.g., a compound, salt, or conjugate and other drugs). The active ingredient may be encapsulated in microcapsules (e.g., hydroxymethylcellulose or gelatin microcapsules and poly(methyl methacrylate) microcapsules, respectively), colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules), or macroemulsions prepared, for example, by coacervation techniques or interfacial polymerization.

[0113] The compositions and formulations may be sterilized. Sterilization may be achieved by filtration by sterile filtration.

[0114] Pharmaceutical compositions containing fast myosin (type II) inhibitors may be formulated for administration as injections. Non-limiting examples of injection preparations may include sterile suspensions, solutions, or emulsions in oily or aqueous media. Suitable oily media may include, but are not limited to, lipophilic solvents or media, such as fatty oils or synthetic fatty acid esters, or liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension. The suspension may also contain suitable stabilizers. Injections may be formulated for bolus injection or continuous infusion. Alternatively, the composition may be lyophilized or in powder form for reconstitution with a suitable vehicle, such as sterile pyrogen-free water, before use.

[0115] Pharmaceutical compositions containing fast myosin (type II) inhibitors may be formulated into injection unit dosage forms (e.g., solutions, suspensions, emulsions) in combination with pharmaceutically acceptable parenteral vehicles. Such vehicles may be essentially non-toxic and non-therapeutic. Vehicles may be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles, such as fixed oils and ethyl oleate, may also be used. Liposomes may be used as carriers. Vehicles may contain small amounts of additives, such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives).

[0116] The pharmaceutical composition comprising a fast-twitch myosin (type II) inhibitor may be formulated for oral delivery to a subject in need thereof.In one embodiment, the composition is formulated to deliver one or more pharmaceutically active agents to a subject through the mucous membrane layer of the oral cavity or esophagus.In another embodiment, the composition is formulated to deliver one or more pharmaceutically active agents to a subject through the mucous membrane layer of the stomach and / or intestine.

[0117] In one embodiment, the composition comprising the fast myosin (type II) inhibitor may comprise a modified release dosage form.Suitable modified release dosage vehicles include, but are not limited to, hydrophilic or hydrophobic matrix devices, water-soluble separating layer coatings, enteric coatings, osmotic devices, multiparticulate devices, and combinations thereof.The composition may also comprise non-release-controlled excipients.

[0118] In another embodiment, the composition containing the fast-twitch myosin (type II) inhibitor may be provided in an enteric-coated dosage form. These enteric-coated dosage forms may also contain non-release-controlled excipients. In one embodiment, the composition is in the form of enteric-coated granules as a controlled-release capsule for oral administration. The composition may further comprise cellulose, disodium hydrogen phosphate, hydroxypropyl cellulose, pyridazine, lactose, mannitol, or sodium lauryl sulfate. In another embodiment, the composition is in the form of enteric-coated pellets as a controlled-release capsule for oral administration. The composition may further comprise glycerol monostearate 40-50, hydroxypropyl cellulose, pyridazine, magnesium stearate, methacrylic acid copolymer type C, polysorbate 80, spherical sugar, talc, or triethyl citrate.

[0119] In another embodiment, the composition comprising a fast-twitch myosin (type II) inhibitor is an enteric-coated controlled-release tablet for oral administration. The composition may further comprise carnauba wax, crospovidone, diacetylated monoglyceride, ethyl cellulose, hydroxypropyl cellulose, pyridazine phthalate, magnesium stearate, mannitol, sodium hydroxide, sodium stearyl fumarate, talc, titanium dioxide, or yellow iron oxide.

[0120] The sustained release composition containing fast myosin (type II) inhibitor can also be prepared.The example of sustained release preparation can include the semipermeable matrix of solid hydrophobic polymer that can contain compound, salt or conjugate, and these matrices can be in the form of shaped article (for example, film or microcapsule).The example of sustained release matrix can include polyester, hydrogel (for example, poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol)), polylactide, L-glutamic acid and γ-ethyl-L-glutamate copolymer, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymer, for example, LUPRON DEPO (i.e., injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(-)-3-hydroxybutyric acid.

[0121] Pharmaceutical compositions containing fast-twitch myosin (type II) inhibitors can be prepared for storage by mixing the compound, salt, or conjugate with a pharmaceutically acceptable carrier, excipient, and / or stabilizer. This formulation can be a lyophilized formulation or an aqueous solution. Acceptable carriers, excipients, and / or stabilizers can be non-toxic to recipients at the dosages and concentrations used. Acceptable carriers, excipients, and / or stabilizers can include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservative polypeptides; proteins such as serum albumin or gelatin; hydrophilic polymers; amino acids; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes; and / or nonionic surfactants or polyethylene glycol.

[0122] Pharmaceutical compositions comprising a fast-twitch myosin (type II) inhibitor can further include calcium stearate, crospovidone, hydroxypropyl methylcellulose, iron oxide, mannitol, methacrylic acid copolymer, polysorbate 80, povidone, propylene glycol, sodium carbonate, sodium lauryl sulfate, titanium dioxide, and triethyl citrate.

[0123] Pharmaceutical compositions containing fast-twitch myosin (type II) inhibitors may be provided in dosage forms having at least one component capable of facilitating immediate release of the active agent and at least one component capable of facilitating controlled release of the active agent. In further embodiments, the dosage form may be capable of providing discontinuous release of the compound in the form of at least two consecutive pulses separated by periods of 0.1 to up to 24 hours. The compositions may include one or more controlled-release and non-controlled-release excipients, such as excipients suitable for disintegrating semipermeable membranes and swellable materials.

[0124] A pharmaceutical composition comprising a fast-twitch myosin (type II) inhibitor is provided in a dosage form for oral administration to a subject, which comprises one or more pharmaceutically acceptable excipients or carriers surrounded by a gastric juice-resistant polymer layered material that is partially neutralized with alkali and has cation exchange capacity, and an intermediate reactive layer that comprises a gastric juice-resistant outer layer.

[0125] Pharmaceutical compositions containing fast myosin (type II) inhibitors provided herein may be in unit-dosage or multi-dosage form. As used herein, unit-dosage form refers to an individually packaged, physically discrete unit suitable for administration to a human or non-human animal subject. Each unit dose may contain a predetermined amount of active ingredient(s) sufficient to produce the desired therapeutic effect, in association with the necessary pharmaceutical carriers or excipients. Examples of unit-dosage forms include, but are not limited to, ampoules, syringes, and individually packaged tablets and capsules. In some embodiments, unit-dosage forms may be administered in fractions or multiples thereof. A multiple-dosage form is a plurality of identical unit-dosage forms packaged in a single container, which may be administered in segregated unit-dosage form. Examples of multiple-dosage forms include, but are not limited to, vials, bottles of tablets or capsules, or bottles of pints or gallons. In another embodiment, the multiple-dosage form contains different pharmaceutically active agents.

[0126] The pharmaceutical composition comprising fast-twitch myosin (type II) inhibitor can also be formulated as modified release dosage form, including immediate release, delayed release, extended release, prolonged release, sustained release, pulsatile release, controlled release, extended release, accelerated and fast release, targeted release, programmed release and gastric retention dosage form.These dosage forms can be prepared according to known methods and techniques (see Remington: The Science and Practice of Pharmacy, supra; Modified-Release Drug Delivery Technology, Rathbone et al., Eds., Drugs and the Pharmaceutical Science, Marcel Dekker, Inc.: New York, NY, 2002; Vol. 126, which are incorporated herein by reference in their entirety). Embodiment

[0127] Embodiment 1. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or increase the subject's North Star Ambulatory Assessment (NSAA) score compared to the NSAA score before treatment.

[0128] Embodiment 2. The method of embodiment 1, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's NSAA score compared to the NSAA score before treatment.

[0129] Embodiment 3. The method of embodiment 2, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's NSAA score within 20% of the pre-treatment NSAA score for a period of 2 months or more, 4 months or more, or 6 months or more.

[0130] Embodiment 4. The method of embodiment 1, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the subject's NSAA score by 0.3 points or more compared to the NSAA score before treatment.

[0131] Embodiment 5. The method of embodiment 4, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the subject's NSAA score by 1 point or more compared to the NSAA score before treatment.

[0132] Embodiment 6. The method of any one of embodiments 1-5, wherein said administering comprises administering a first dose of said fast-twitch myosin (type II) inhibitor and one or more subsequent doses of said fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0133] Embodiment 7. The method of embodiment 6, wherein the subject's NSAA score is assessed 2 months or more, 4 months or more, or 6 months or more after the subject's first dose of said fast-twitch myosin (type II) inhibitor.

[0134] Embodiment 8. The method of embodiment 6 or 7, wherein the pre-treatment NSAA score is assessed within 1 month prior to the subject's first dose of said fast-twitch myosin (type II) inhibitor.

[0135] Embodiment 9. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or improve one or more of the following functional measures in the subject: maximal elbow flexion strength compared to maximal elbow flexion strength before treatment; maximal knee extension strength compared to maximal knee extension strength before treatment; 10 meter walk / run speed compared to 10 meter walk / run speed before treatment; 100 meter timed test speed compared to 100 meter timed test speed before treatment; 4-step stair climbing timed speed compared to 4-step stair climbing timed speed before treatment; and maximal grip strength compared to maximal grip strength value before treatment.

[0136] Embodiment 10. 100 meter timed test speed 100 meter timed test speed Timed speed of climbing 4 steps Timed speed of climbing 4 steps. The method of embodiment 9, comprising administering an amount of a fast-twitch myosin (type II) inhibitor sufficient to maintain one or more functional measures of the subject compared to pre-treatment values.

[0137] Embodiment 11. The method of embodiment 10, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain one or more functional measures of the subject within 20% of pre-treatment values ​​for a period of 2 months or more, 4 months or more, or 6 months or more.

[0138] Embodiment 12. The method of embodiment 9, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase one or more functional measures in the subject by 20% or greater compared to pre-treatment values.

[0139] Embodiment 13. The method of embodiment 12, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase one or more functional measures in the subject by 30% or greater compared to pre-treatment values.

[0140] Embodiment 14. The method of any one of embodiments 9-13, wherein said administering comprises administering a first dose of said fast-twitch myosin (type II) inhibitor and one or more subsequent doses of said fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0141] Embodiment 15. The method of embodiment 14, wherein the subject's one or more functional measures are assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0142] Embodiment 16. The method of embodiment 14 or 15, wherein the pre-treatment value is assessed within one month prior to the subject's first dose of said fast-twitch myosin (type II) inhibitor.

[0143] Embodiment 17. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or reduce the subject's creatine kinase activity level by 10% or more compared to the creatine kinase activity level before treatment.

[0144] Embodiment 18. The method of embodiment 17, comprising administering an amount of a fast-twitch myosin (type II) inhibitor sufficient to maintain creatine kinase activity in the subject compared to pre-treatment creatine kinase activity levels.

[0145] Embodiment 19. The method of embodiment 18, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's creatine kinase activity level within 10% of the pre-treatment creatine kinase activity level.

[0146] Embodiment 20. The method of embodiment 17, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the subject's creatine kinase activity level by 15% or more, 20% or more, 25% or more, 30% or more, or 35% or more compared to the creatine kinase activity level before treatment.

[0147] Embodiment 21. The method of any one of embodiments 17-20, wherein said administering comprises administering a first dose of said fast-twitch myosin (type II) inhibitor and one or more subsequent doses of said fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0148] Embodiment 22. The method of embodiment 21, wherein the subject's creatine kinase activity level is assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0149] Embodiment 23. The method of embodiment 21 or 22, wherein the pre-treatment creatine kinase activity level is assessed within one month prior to administering a first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0150] Embodiment 24. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain serum creatinine levels or to increase the subject's serum creatinine levels by 10% or more compared to the serum creatinine level before treatment.

[0151] Embodiment 25. The method of embodiment 24, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's serum creatinine level compared to the serum creatinine level before treatment.

[0152] Embodiment 26. The method of embodiment 25, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's serum creatinine level within 10% of the pre-treatment serum creatinine level for a period of 2 months or more, 4 months or more, or 6 months or more.

[0153] Embodiment 27. The method of embodiment 24, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the subject's serum creatinine level by 10% or more, or 20% or more, compared to the serum creatinine level before treatment.

[0154] Embodiment 28. The method of any one of embodiments 24-27, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0155] Embodiment 29. The method of embodiment 28, wherein the subject's serum creatinine level is assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0156] Embodiment 30. The method of embodiment 28 or 29, wherein the pre-treatment serum creatinine level is assessed within 1 month prior to the subject's first dose of said fast-twitch myosin (type II) inhibitor.

[0157] Embodiment 31. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's dual-energy x-ray absorptiometry (DXA) % lean mass or to increase the subject's DXA % lean mass by 5% or more compared to the DXA % lean mass before treatment.

[0158] Embodiment 32. The method of embodiment 31, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's DXA % lean mass compared to pre-treatment DXA % lean mass.

[0159] Embodiment 33. The method of embodiment 32, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's DXA lean mass % within 5% of the pre-treatment DXA lean mass % for a period of 2 months or more, 4 months or more, or 6 months or more.

[0160] Embodiment 34. The method of embodiment 31, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the subject's DXA % lean mass by 5% or more, 10% or more, or 15% or more compared to the DXA % lean mass before treatment.

[0161] Embodiment 35. The method of any one of embodiments 31-34, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0162] Embodiment 36. The method of embodiment 35, wherein the subject's DXA % lean mass is assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0163] Embodiment 37. The method of embodiment 35 or 36, wherein the pre-treatment DXA lean mass % is assessed within 1 month prior to the subject's first dose of said fast-twitch myosin (type II) inhibitor.

[0164] Embodiment 38. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's fast skeletal troponin I (TNNI2) level or reduce the subject's TNNI2 level by 10% or more compared to the TNNI2 level before treatment, wherein optionally the TNNI2 level is estimated from SOMAscan levels.

[0165] Embodiment 39. The method of embodiment 38, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain a TNNI2 level in the subject compared to the TNNI2 level before treatment.

[0166] Embodiment 40. The method of embodiment 39, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's TNNI2 concentration within 10% of pre-treatment creatine kinase activity level.

[0167] Embodiment 41. The method of embodiment 38, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the subject's TNNI2 concentration by 15% or more, 20% or more, 25% or more, 30% or more, 40% or more, 50% or more, 60% or more, or 70% or more compared to the TNNI2 concentration before treatment.

[0168] Embodiment 42. The method of any one of embodiments 38-41, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0169] Embodiment 43. The method of embodiment 42, wherein the subject's TNNI2 concentration is assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0170] Embodiment 44 The method of embodiment 42 or 43, wherein the TNNI2 concentration is assessed within one month prior to the first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0171] Embodiment 45. The fast myosin (type II) inhibitor is a compound of formula (I): [ka] 10. The method of any one of the preceding embodiments, wherein

[0172] Embodiment 46. The method of any one of the preceding embodiments, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor once daily, twice daily, three times daily, once every two days, once every three days, once every four days, once every five days, once every six days, or once weekly.

[0173] Embodiment 47. The method of embodiment 46, wherein administering comprises administering a fast-twitch myosin (type II) inhibitor once daily.

[0174] Embodiment 48. The method of any one of embodiments 1-47, wherein administering comprises administering to the subject a fast-twitch myosin (type II) inhibitor for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0175] Embodiment 49. The method of embodiment 48, wherein administering comprises administering to the subject a fast-twitch myosin (type II) inhibitor for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0176] Embodiment 50. The method of any preceding embodiment, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0177] Embodiment 51. The method of embodiment 50, wherein the neuromuscular condition is Becker muscular dystrophy.

[0178] Embodiment 52. The method of embodiment 50, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0179] Embodiment 53 The method of any one of the preceding embodiments, wherein the subject is 18 years of age or younger.

[0180] Embodiment 54. The method of any one of embodiments 1 to 52, wherein the subject is over 18 years of age.

[0181] Embodiment 55. Prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a) to (g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a pre-treatment serum lipid profile of -0.1 to -10, or -0.5 to -3. The method of any preceding embodiment, further comprising selecting subjects for treatment having one or more of: (a) NSAA annual change; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) a self-reported pain score associated with muscular dystrophy of 1.5 or greater, or 2 or greater.

[0182] Embodiment 56. A method of inhibiting fast-twitch myosin (type II) in a subject in need thereof, comprising the steps of: (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a dystrophin gene mutation in the blood; (e) a dystrophin gene mutation in the blood; (f) a dystrophin gene mutation in the blood; (g) a dystrophin gene mutation in the blood; (h) a dystrophin gene mutation in the blood; (i) a dystrophin gene mutation in the blood; (j ... (e) pre-treatment NSAA annual change of -1 to -4; (e) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (f) DXA lean mass % of <75%, <70%, <65%, <60%; and (g) self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher; and administering to the subject a fast-twitch myosin (type II) inhibitor.

[0183] Fast Myosin (Type II) Embodiment 57. The method of embodiment 56, wherein the dystrophin gene mutation is selected from del 45-48, del 45-47, del 48, del x-51, del 45-55, and del 48-49.

[0184] Embodiment 58. The fast myosin (type II) inhibitor is a compound of formula (I): [ka] 58. The method of embodiment 56 or 57, wherein

[0185] Embodiment 59. The method of any one of the preceding embodiments, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor once daily, twice daily, three times daily, once every two days, once every three days, once every four days, once every five days, once every six days, or once weekly.

[0186] Embodiment 60. The method of embodiment 59, wherein administering comprises administering a fast-twitch myosin (type II) inhibitor once daily.

[0187] Embodiment 61. The method of any one of embodiments 56-60, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0188] Embodiment 62. The method of embodiment 61, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0189] Embodiment 63. The method of any one of embodiments 56-62, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0190] Embodiment 64. The method of embodiment 63, wherein the neuromuscular condition is selected Becker muscular dystrophy.

[0191] Embodiment 65. The method of embodiment 63, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0192] Embodiment 66. The method of any one of embodiments 56 to 65, wherein the subject is 18 years of age or younger.

[0193] Embodiment 67. The method of any one of embodiments 56-66, wherein administering does not negatively affect cardiac function of the subject.

[0194] Embodiment 68. The method of any one of embodiments 61 to 67, wherein one or more of characteristics (a) to (g) is improved after said administering.

[0195] Embodiment 69. A compound of Formula (I) in an amount of about 1 mg to about 20 mg [ka] and a pharmaceutically acceptable excipient.

[0196] Embodiment 70. The unit dose of embodiment 69, comprising about 1 mg to about 5 mg of a compound of Formula (I), and a pharmaceutically acceptable excipient.

[0197] Embodiment 71. The unit dose of embodiment 70, comprising about 1 mg to about 3 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0198] Embodiment 72. The unit dose of embodiment 70, comprising about 2.5 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0199] Embodiment 73. The unit dose according to any one of embodiments 69 to 72, wherein the unit dose is formulated as a pill or tablet.

[0200] Embodiment 74. A kit comprising an oral pharmaceutical composition and instructions for administration to a subject in need thereof, wherein the oral pharmaceutical composition comprises a compound of formula (I): [ka] and a pharmaceutically acceptable excipient.

[0201] Embodiment 75. A method of treating a neuromuscular condition, comprising administering to a subject in need thereof a compound of formula (I) at a dose of 3 mg to 40 mg per day: [ka] The method of claim 1, wherein the

[0202] Embodiment 76 The method of embodiment 75, comprising administering to the subject a dose of 15 mg to 40 mg per day of the compound of formula (I).

[0203] Embodiment 77. The method of embodiment 76, comprising administering to the subject a dose of 15 mg, 20 mg, 25 mg, or 30 mg per day of the compound of formula (I).

[0204] Embodiment 78. The method of embodiment 77, comprising administering to the subject a dose of 20 mg per day of the compound of formula (I).

[0205] Embodiment 79. The method of embodiment 75, comprising administering to the subject a dose of 2 mg to 15 mg per day of the compound of formula (I).

[0206] Embodiment 80. The method of embodiment 79, comprising administering to the subject a dose of 5 mg to 12.5 mg per day of the compound of formula (I).

[0207] Embodiment 81 The method of embodiment 80, comprising administering to the subject a dose of 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, or 12.5 mg per day of the compound of formula (I).

[0208] Embodiment 82. The method of any one of embodiments 75 to 81, wherein the subject is over 18 years of age.

[0209] Embodiment 83. The method of any one of embodiments 75 to 81, wherein the subject is 18 years of age or younger.

[0210] Embodiment 84. The method of any of embodiments 75-83, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0211] Embodiment 85. The method of embodiment 84, wherein the neuromuscular condition is Becker muscular dystrophy.

[0212] Embodiment 86. The method of embodiment 84, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0213] Embodiment 87. The method of any one of embodiments 75-86, wherein administering comprises administering the compound of formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0214] Embodiment 88. The method of embodiment 87, wherein administering comprises administering the compound of formula (I) for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0215] Embodiment 89. The method of embodiment 88, wherein administering comprises administering the compound of formula (I) for 6 months or longer.

[0216] Embodiment 90. A method of treating a neuromuscular condition, comprising administering to a subject an AUC 24 a compound of formula (I) in an amount sufficient to maintain [ka] The method of claim 1, wherein the

[0217] Embodiment 91. The method of embodiment 90, wherein administering comprises administering the compound of formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0218] Embodiment 92. The method of embodiment 91, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0219] Embodiment 93. A compound of formula (I) is administered to a subject in need thereof, wherein the AUC 24 The method of any one of embodiments 90-92, wherein the method is administered in an amount sufficient to maintain

[0220] Embodiment 94. A compound of formula (I) is administered to a subject in need thereof, wherein the AUC 2494. The method of embodiment 93, wherein the patient is administered in an amount sufficient to maintain

[0221] Embodiment 95. The method of any one of embodiments 90-94, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0222] Embodiment 96. The method of embodiment 95, wherein the neuromuscular condition is Becker muscular dystrophy.

[0223] Embodiment 97. The method of embodiment 95, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0224] Embodiment 98. The method of any one of embodiments 90 to 97, wherein the subject is over 18.

[0225] Embodiment 99. The method of any one of embodiments 90 to 97, wherein the subject is 18 years of age or younger.

[0226] Embodiment 100. The method of any one of embodiments 90-99, wherein administering does not negatively affect cardiac function of the subject.

[0227] Embodiment 101. The method of any one of embodiments 90-100, wherein administering comprises administering the compound of formula (I) once a day, twice a day, three times a day, once every two days, once every three days, once every four days, once every five days, once every six days, or once a week.

[0228] Embodiment 102. The method of embodiment 101, wherein administering comprises administering the compound of formula I once daily.

[0229] Embodiment 103. Prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a) to (g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a pre-treatment N of -0.1 to -10, or -0.5 to -3. The method of any one of embodiments 90-102, further comprising selecting subjects for treatment having one or more of: (a) SAA annual change; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) a self-reported pain score associated with muscular dystrophy of 1.5 or greater, or 2 or greater.

[0230] Embodiment 104. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain or reduce the subject's myoglobin level by 10% or more compared to the myoglobin level before treatment.

[0231] Embodiment 105. The method of embodiment 104, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's myoglobin level compared to the myoglobin level before treatment.

[0232] Embodiment 106. The method of embodiment 104, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to maintain the subject's myoglobin level within 10% of the pre-treatment myoglobin level.

[0233] Embodiment 107. The method of embodiment 104, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the subject's myoglobin level by 5% or more, 10% or more, 15% or more, 20% or more, or 25% or more compared to the myoglobin before treatment.

[0234] Embodiment 108. The method of any one of embodiments 104 to 107, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0235] Embodiment 109. The method of any one of embodiments 104-108, wherein the subject's myoglobin level is assessed 2 months or more, 4 months or more, or 6 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0236] Embodiment 110. The method of any one of embodiments 104-109, wherein the pre-treatment myoglobin level is assessed within one month prior to administering a first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0237] Embodiment 111. A method of reducing pain associated with a neuromuscular condition, comprising administering to a subject in need thereof a fast-twitch myosin (type II) inhibitor.

[0238] Embodiment 112. The method of embodiment 111, wherein the pain is diffuse.

[0239] Embodiment 113. The method of embodiment 112, wherein the pain is in the spine and calf of the subject.

[0240] Embodiment 114. The method of any of embodiments 111-113, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0241] Embodiment 115. The method of embodiment 114, wherein the neuromuscular condition is Becker muscular dystrophy.

[0242] Embodiment 116. The method of embodiment 114, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0243] Embodiment 117. The method of any of the preceding embodiments, wherein the fast-twitch myosin (type II) inhibitor is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0244] Embodiment 118. The method of embodiment 117, wherein the fast-twitch myosin (type II) inhibitor is administered at a constant dose throughout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0245] Embodiment 119. The method of embodiment 118, wherein the constant dose is 10 mg / day, 15 mg / day, or 20 mg / day.

[0246] Embodiment 120. The method of embodiment 119, wherein the constant dose is 10 mg / day.

[0247] Embodiment 121. The method of embodiment 117, wherein the fast-twitch myosin (type II) inhibitor is administered at various doses throughout 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0248] Embodiment 122. The method of embodiment 121, wherein the fast-twitch myosin (type II) inhibitor is administered in decreasing doses over 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0249] Embodiment 123. The method of embodiment 121, wherein the fast-twitch myosin (type II) inhibitor is administered in escalating doses over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0250] Embodiment 124. The method of embodiment 123, wherein the fast-twitch myosin (type II) inhibitor is administered at 10 mg / day, followed by 15 mg / day, and further followed by 20 mg / day.

[0251] Embodiment 125. The method of embodiment 124, wherein the fast-twitch myosin (type II) inhibitor is administered at 10 mg / day in months 1 and 2, followed by 15 mg / day in months 3 through 6, and further followed by 20 mg / day in months 7 through 12.

[0252] Embodiment 126. The method of embodiment 121, wherein the fast-twitch myosin (type II) inhibitor is administered at varying doses to maintain a plasma concentration of the fast-twitch myosin (type II) inhibitor in the subject between 10 ng / ml and 100 ng / ml, between 15 and 70 ng / ml, between 20 ng / ml and 70 ng / ml, or between 30 ng / ml and 70 ng / ml.

[0253] Embodiment 127. The method of embodiment 121, wherein the fast-twitch myosin (type II) inhibitor is administered at varying doses to maintain a muscle concentration of the fast-twitch myosin (type II) inhibitor in the subject of 1,000 to 4,000 ng / g.

[0254] Embodiment 128. The method of any one of the preceding embodiments, wherein the fast-twitch myosin (type II) inhibitor is enriched in muscle.

[0255] Embodiment 129. The fast myosin (type II) inhibitor is a compound of formula (I): [ka] 10. The method of one of the preceding embodiments, comprising:

[0256] Embodiment 130. The unit dose of embodiment 69, comprising about 1 mg to about 15 mg of a compound of Formula (I), and a pharmaceutically acceptable excipient.

[0257] Embodiment 131. The unit dose of embodiment 69, comprising about 5 mg to about 15 mg of a compound of Formula (I), and a pharmaceutically acceptable excipient.

[0258] Embodiment 132. The unit dose of embodiment 69, comprising about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 12.5 mg, about 15 mg, about 17.5 mg, or about 20 mg of a compound of Formula (I), and a pharmaceutically acceptable excipient.

[0259] Embodiment 133. The unit dose of embodiment 69, comprising about 2.5 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0260] Embodiment 134. The unit dose of embodiment 69, comprising about 5 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0261] Embodiment 135. The unit dose of embodiment 69, comprising about 10 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0262] Embodiment 136 The method of embodiment 75, comprising administering to the subject a dose of about 1 mg to about 15 mg per day of a compound of formula (I).

[0263] Embodiment 137. The method of embodiment 75, comprising administering to the subject a dose of about 5 mg to about 15 mg per day of a compound of Formula (I).

[0264] Embodiment 138. The method of embodiment 75, comprising administering to the subject a dose of about 2.5 mg per day, about 5 mg per day, about 7.5 mg per day, about 10 mg per day, about 12.5 mg per day, about 15 mg per day, about 17.5 mg per day, or about 20 mg per day of a compound of Formula (I).

[0265] Embodiment 139. The method of embodiment 75, comprising administering to the subject a dose of about 2.5 mg per day of the compound of formula (I).

[0266] Embodiment 140. The method of embodiment 75, comprising administering to the subject a dose of about 5 mg per day of a compound of formula (I).

[0267] Embodiment 141. The method of embodiment 75, comprising administering to the subject a dose of about 10 mg per day of the compound of formula (I).

[0268] Embodiment 142. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more compared to the pre-treatment protein concentration of the one or more muscle injury biomarkers.

[0269] Embodiment 143. The method of embodiment 142, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more muscle injury biomarkers in the subject to within 10% of the pre-treatment protein concentration of the one or more muscle injury biomarkers.

[0270] Embodiment 144. The method of embodiment 142, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more muscle injury biomarkers in the subject by 15% or more, 20% or more, 25% or more, 30% or more, or 35% or more compared to the pre-treatment protein concentration of the one or more muscle injury biomarkers.

[0271] Embodiment 145. The method of any one of embodiments 142 to 144, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0272] Embodiment 146. The method of embodiment 145, wherein the protein concentration of the one or more muscle injury biomarkers in the subject is assessed 2 months or more, 4 months or more, 6 months or more, 8 months or more, 10 months or more, or 12 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0273] Embodiment 147. The method of embodiment 145 or 146, wherein the protein concentration of one or more muscle injury biomarker levels is assessed within one month prior to the first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0274] Embodiment 148. The method of any one of embodiments 142 to 147, wherein the one or more muscle injury biomarkers comprise ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof.

[0275] Embodiment 149. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more compared to the pre-treatment protein concentration of the one or more pro-inflammatory proteins.

[0276] Embodiment 150. The method of embodiment 149, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more pro-inflammatory proteins in the subject to within 10% of the pre-treatment protein concentration of the one or more pro-inflammatory proteins.

[0277] Embodiment 151. The method of embodiment 149, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to reduce the protein concentration of one or more pro-inflammatory proteins in the subject by 15% or more, 20% or more, 25% or more, 30% or more, or 35% or more compared to the pre-treatment protein concentration of the one or more pro-inflammatory proteins.

[0278] Embodiment 152. The method of any one of embodiments 149 to 151, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0279] Embodiment 153. The method of embodiment 152, wherein the protein concentration of the one or more pro-inflammatory proteins in the subject is assessed 2 months or more, 4 months or more, 6 months or more, 8 months or more, 10 months or more, or 12 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0280] Embodiment 154. The method of embodiment 152 or 153, wherein the protein concentration of one or more pro-inflammatory proteins is assessed within one month prior to the first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0281] Embodiment 155. The method of any one of embodiments 149-154, wherein the one or more pro-inflammatory proteins comprise CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof.

[0282] Embodiment 156. A method of treating a neuromuscular condition, comprising administering to a subject a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more compared to the protein concentration of the one or more anti-inflammatory proteins before treatment.

[0283] Embodiment 157. The method of embodiment 156, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the protein concentration of the one or more anti-inflammatory proteins in the subject to within 10% of the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

[0284] Embodiment 158. The method of embodiment 156, comprising administering a fast-twitch myosin (type II) inhibitor in an amount sufficient to increase the protein concentration of one or more anti-inflammatory proteins in the subject by 15% or more, 20% or more, 25% or more, 30% or more, or 35% or more compared to the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

[0285] Embodiment 159. The method of any one of embodiments 156 to 158, wherein the administering comprises administering a first dose of the fast-twitch myosin (type II) inhibitor and one or more subsequent doses of the fast-twitch myosin (type II) inhibitor, optionally wherein the one or more subsequent doses are higher than the first dose.

[0286] Embodiment 160. The method of embodiment 159, wherein the protein concentration of the one or more anti-inflammatory proteins in the subject is assessed 2 months or more, 4 months or more, 6 months or more, 8 months or more, 10 months or more, or 12 months or more after a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0287] Embodiment 161. The method of embodiment 159 or 160, wherein the protein concentration of one or more anti-inflammatory proteins is assessed within one month prior to the first dose of said fast-twitch myosin (type II) inhibitor to the subject.

[0288] Embodiment 162. The method of any one of embodiments 156 to 161, wherein the one or more anti-inflammatory proteins comprise IL10, IL13, IL22, IL37, IL4, or a combination thereof.

[0289] Embodiment 163. The fast myosin (type II) inhibitor is a compound of formula (I): [ka] The method of any one of embodiments 142 to 162, wherein

[0290] Embodiment 164. The method of any one of embodiments 142-163, wherein administering comprises administering the fast-twitch myosin (type II) inhibitor once a day, twice a day, three times a day, once every two days, once every three days, once every four days, once every five days, once every six days, or once a week.

[0291] Embodiment 165. The method of embodiment 164, wherein administering comprises administering a fast-twitch myosin (type II) inhibitor once daily.

[0292] Embodiment 166. The method of any one of embodiments 142-165, wherein administering comprises administering to the subject a fast-twitch myosin (type II) inhibitor for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0293] Embodiment 167. The method of embodiment 166, wherein administering comprises administering to the subject a fast-twitch myosin (type II) inhibitor for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0294] Embodiment 168. The method of any one of embodiments 142 to 167, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0295] Embodiment 169. The method of embodiment 168, wherein the neuromuscular condition is Becker muscular dystrophy.

[0296] Embodiment 170. The method of embodiment 168, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0297] Embodiment 171. The method of any one of embodiments 142 to 170, wherein the subject is 18 years of age or younger.

[0298] Embodiment 172. The method of any one of embodiments 142 to 170, wherein the subject is over 18 years of age.

[0299] Embodiment 173. Prior to said administering, the method further comprises detecting a patient having one of the following characteristics (a) to (g): (a) a creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) a dystrophin gene mutation; (d) a pre-treatment NS of -0.1 to -10, or -0.5 to -3. The method of any one of embodiments 142-172, further comprising selecting subjects for treatment having one or more of: (a) AA annual change; (b) pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (c) DXA lean mass % of <75%, <70%, <65%, <60%; and (d) a self-reported pain score associated with muscular dystrophy of 1.5 or higher, or 2 or higher. [Example]

[0300] The invention now being generally described, this will be more readily understood by reference to the following examples, which are included solely for purposes of illustration of certain aspects and embodiments of the invention and are not intended to limit the invention in any way. Example 1 Human Clinical Trials

[0301] Figures 2-8 present positive 4-month interim results from an ongoing, open-label, single-center study evaluating the safety, tolerability, and effects on muscle damage biomarkers, as well as pharmacokinetics (PK), of Compound 1 in adults with BMD. Twelve adults with BMD enrolled in the ARCH study initially received a 10 mg dose during the first 2 months of the study, followed by dose escalation to a daily 15 mg oral dose of Compound 1 at night. Compound 1 was well tolerated in all participants, with no discontinuations or dose reductions. The most common adverse events observed to date were dizziness, somnolence, and headache. All eligible patients subsequently had their dose increased to 20 mg daily, per protocol.

[0302] Treatment with Compound 1 resulted in significant reductions in key biomarkers of muscle damage, as assessed by laboratory assays. Importantly, CK and fast skeletal troponin I were reduced by an average of 29% and 73%, respectively, after 4 months (Figure 5). CK reduction was sustained and consistent with observations made at 2 months, while fast skeletal troponin I levels continued to decline with continued exposure to Compound 1, with four patients reaching levels observed in unaffected adults. Similar to observations made at 2 months, both CK and fast skeletal troponin I were significantly reduced under typical daily activity levels measured by pedometer.

[0303] Four months after Compound 1 administration, NSAA increased by a mean of 1.17 points compared to pretreatment baseline. Notably, 9 of 12 participants showed either functional improvement or no decline in NSAA compared to their baseline (Figure 2). The NSAA improvement observed after only 4 months of Compound 1 administration differs from the trajectory observed in a natural history study reported by Bello et al. (2016), one of the most comprehensively characterized BMD cohorts, in which the annual decline was 1.22 NSAA points. These observations were further corroborated by an independent study from van de Velde et al. (2021), which showed a 2.5 NSAA point decline over 2 years. Example 2 Ongoing human clinical trials

[0304] Figures 10-13 present promising 12-month results from an ongoing, open-label, single-center study evaluating the safety, tolerability, and effects on muscle damage biomarkers, as well as pharmacokinetics (PK), of Compound 1 in adults with BMD. Different dosing regimens, depicted in Figures 10A-10C, 11B, and 12A-12F, were administered to three cohorts. One cohort received 20 mg of Compound 1 for 12 months. The second cohort received 15 mg of Compound 1 for 6 months. The third cohort received 10 mg of Compound 1 for 2 months. Compound 1 was well tolerated at all doses without any serious adverse events (AEs). Therefore, the treatment regimen was not discontinued due to AEs.

[0305] Several muscle damage biomarkers were tested to evaluate the efficacy of Compound 1 with different dosing regimens. Figures 10A-10C illustrate that Compound 1 resulted in sustained reductions in creatine kinase (CK), fast skeletal troponin I (TNNI2), and myoglobin. Notably, the 10 mg dose showed near-maximal reductions. When analyzing biomarker changes in individual participants from the 12-month group, individuals with the highest baseline values ​​of CK or TNNI2 showed the greatest reductions in biomarkers with Compound 1, suggesting that Compound 1 provided protection against activity-induced damage. See Figures 10D-10G.

[0306] NSAA scores were also quantified based on Figures 9A-9C. Compound 1 stabilized NSAA scores by rescuing the natural history trend by approximately 1.6 points, resulting in a trend toward score improvement. See Figure 11A. Individual NSAA responses from the 12-month group are reflected in Figure 11C, which showed that approximately 75% of participants from the 12-month group had the same or better NSAA scores. In comparison, the expected natural history decline in untreated patients is approximately -1.2 to -1.3 points.

[0307] In parallel, plasma concentrations of Compound 1 were also monitored across different dosing regimens. See Figure 11B. For higher dose groups (e.g., the 20 mg group), there were patients with high or low exposure. It was unexpected that high-exposure patients reached a range of plasma concentrations after 20 mg / day dosing that was more than four-fold higher than the target exposure predicted from preclinical models. It is also noteworthy that Compound 1 was abundant in muscle. The target muscle concentrations of Compound 1 from patients with lower plasma concentrations ranged from 1,000 to 4,000 ng / g in preclinical experiments. Using muscle biopsy data from a Phase 1 multiple-ascending dose (MAD) study, this equates to approximately 15 to 70 ng / ml in BMD. In other words, Compound 1 was approximately 60-fold more abundant in muscle than in plasma.

[0308] Furthermore, participants from the lower exposure 20 mg group (highlighted by the lower circle in Figure 11D) had stable or best improvement in NSAA score (see the dark shaded bar in Figure 11C). In contrast, participants with less optimal results (e.g., negative change in NSAA score in Figure 11C) were under high exposure of Compound 1 (highlighted by the upper circle in Figure 11D). Therefore, lower exposure of Compound 1 (e.g., 10 mg) may have more desirable functional benefits.

[0309] Furthermore, functional tests were carried out to evaluate the muscle function improvement in participants resulting from Compound 1. For example, no statistically significant deterioration was observed in 10m speed (see Figure 12A), 100m speed (see Figure 12B), or 4-4 stair climbing time speed (see Figure 12C). Different parameters of muscle strength were also quantified and evaluated in Figures 12D-12F. Furthermore, it was observed that some participants from the 20mg group showed muscle strength weakness (see, for example, Figures 12C-12F).

[0310] In combination with the data shown in Figures 10A-10C and 11B-11D, improvement of activity-induced muscle damage and reduction of related biomarkers indicating a good NSAA score could be achieved at 10 mg / day. A dose of 20 mg / day produced efficacy, however, some muscle weakness also occurred. Therefore, optimally controlled dosage (e.g., 10 mg / day) of Compound 1 could produce a well-balanced result of muscle preservation and avoidance of some muscle weakness.

[0311] Individuals with BMD typically report more diffuse pain in the spine and calf (see Jacques MF et al., Plos ONE (2019)). Self-reported pain assessments were collected and quantified in Figure 13. A positive trend in self-reported pain scores was observed 12 months after Compound 1 treatment. Thus, Compound 1 was shown to reduce pain associated with BMD. Example 3 Characterization of short- and long-term proteomic responses to fast skeletal myosin inhibitors in Becker muscular dystrophy (BMD)

[0312] Changes in muscle injury biomarkers, such as creatine kinase (CK) or fast skeletal troponin I (TNNI2), were assessed in clinical trial participants. Biomarker abundance was measured by plasma proteomics (SOMAscan) during short-term (1-2 months), intermediate-term (3-4 months), or long-term (6-12 months) treatment. Figure 14A illustrates the study design. Twelve BMD participants (seven of whom participated in the Phase I MAD study (Example 2)) were enrolled and initially treated with 10 mg of Compound 1 daily, with dose escalation from 10 mg to 20 mg daily as indicated. Blood samples were collected at baseline before dosing and at regular intervals thereafter for analysis by SOMAscan and quantification of CK activity or TNNI2 concentration by activity assay or ELISA. SOMAscan is a multistep, engineered aptamer-based assay for high-throughput, sensitive, and unbiased biomarker measurement. Proteins in the sample were selectively bound to the fluorescent aptamers, which were then measured in the chip array to give values ​​proportional to absolute concentration.

[0313] In this study, SOMAscan not only correlated with CK and TNNI2, allowing direct conversion to absolute concentrations, but was also used to identify large-scale proteomic differences between short-term and long-term treatment with Compound 1. Figure 14B illustrates the validation of CK and TNNI2, where a strong correlation was observed between SOMAscan and absolute measurements of CK (left) and TNNI2 (right). Figure 14C illustrates CK and TNNI2 quantified by SOMAscan in blood. Circulating CK and TNNI2 were significantly increased above values ​​seen in healthy individuals at baseline and reduced by short-term treatment. TNNI2 was reduced to near-healthy levels at longer-term treatment. The significance shown was present for all values ​​at that treatment time point compared to the pre-dose baseline. Figures 14D and 14E illustrate the effect of Compound 1 on muscle injury proteins. SOMAscan was used to analyze the effects of treatment on a panel of proteins identified as correlated with bona fide contraction-induced skeletal muscle injury (Figure 14D). These proteins, such as CK and TNNI2, were reduced from pre-dose baseline with short-term treatment and showed stable levels or further reductions over the course of long-term treatment. As a group, there was no difference in muscle injury protein responses between short-term and long-term treatment (Figure 14E), indicating that the muscle effects of treatment were maximized by 1-2 months of 10 mg Compound 1 treatment.

[0314] Figure 14F illustrates the proteomics of long-term Compound 1 treatment. Proteins that were decreased by long-term treatment or differentiated based on long-term treatment compared to short-term treatment showed two distinct clusters of changes. Cluster A was characterized by several skeletal muscle proteins that were significantly decreased from the pre-treatment baseline after both short-term and long-term treatment. In contrast, proteins in Cluster B were only responsive after a longer period of Compound 1 treatment. These proteins primarily represented immune and inflammatory pathways.

[0315] Figures 14G and 14H illustrate favorable shifts in inflammatory biomarkers with long-term treatment. Favorable shifts were observed in the inflammatory profiles of study participants after long-term Compound 1 treatment. Several pro-inflammatory cytokines and chemokines were highly significantly reduced after 6 to 12 months, as indicated by a decrease in grayscale intensity (upper panel of Figure 14G), while several anti-inflammatory interleukins increased after long-term treatment, as indicated by an increase in grayscale intensity (lower panel of Figure 14G). The observed protein concentration changes after 6 to 12 months were consistent across study participants (Figure 14H).

[0316] As shown in this study, SOMAscan values ​​strongly correlated with absolute measurements of CK and TNNI2. CK, TNNI2, and a broad set of muscle injury proteins were elevated at pretreatment baseline, decreased soon after treatment began, and remained significantly reduced with long-term treatment. Short-term proteomics of Compound 1 treatment were dominated by skeletal muscle proteins reflecting muscle damage. In addition, long-term treatment also showed a consistent shift in proteins associated with inflammation, where pro-inflammatory factors were reduced while anti-inflammatory cytokines were increased. While this study varied both time and exposure, these proteins would be expected to require longer periods to confirm significant changes, which may reflect muscle normalization in the sustained reduction of muscle damage. The proteomic profile of Compound 1 treatment showed a rapid and sustained decrease in markers of muscle injury, accompanied by a longer-term change in the inflammatory milieu, shifting the proteomic profile toward that of healthy individuals.

[0317] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will occur to those skilled in the art herein without departing from the invention. It should be understood that various modifications to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention, and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. A method of treating a neuromuscular condition, comprising administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day: 【Chemistry 18】 The method of claim 1, wherein the

2. 10. The method of claim 1, comprising administering to the subject a dose of 15 mg to 40 mg of the compound of formula (I) per day.

3. 3. The method of claim 2, comprising administering to the subject a dose of 15 mg, 20 mg, 25 mg, or 30 mg of the compound of formula (I) per day.

4. 4. The method of claim 3, comprising administering to the subject a dose of 20 mg per day of the compound of formula (I).

5. 10. The method of claim 1, comprising administering to the subject a dose of the compound of formula (I) of 1 mg to 20 mg per day.

6. 6. The method of claim 5, comprising administering to the subject a dose of 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, or 20 mg of the compound of formula (I) per day.

7. 7. The method of claim 6, comprising administering to the subject a dose of 1 mg per day of the compound of formula (I).

8. 7. The method of claim 6, comprising administering to the subject a dose of 2 mg per day of the compound of formula (I).

9. 7. The method of claim 6, comprising administering to the subject a dose of 2.5 mg per day of the compound of formula (I).

10. 7. The method of claim 6, comprising administering to the subject a dose of 5 mg per day of the compound of formula (I).

11. 7. The method of claim 6, comprising administering to the subject a dose of 7.5 mg per day of the compound of formula (I).

12. 7. The method of claim 6, comprising administering to the subject a dose of 10 mg per day of the compound of formula (I).

13. 7. The method of claim 6, comprising administering to the subject a dose of 15 mg per day of the compound of formula (I).

14. 7. The method of claim 6, comprising administering to the subject a dose of 20 mg per day of the compound of formula (I).

15. The method of any one of claims 1 to 14, wherein the subject is over 18 years of age.

16. 16. The method of claim 15, comprising administering to the subject a dose of 10 mg per day of the compound of formula (I).

17. 16. The method of claim 15, comprising administering to the subject a dose of 15 mg per day of the compound of formula (I).

18. 16. The method of claim 15, comprising administering to the subject a dose of 20 mg per day of the compound of formula (I).

19. The method of any one of claims 1 to 7, wherein the subject is 18 years of age or younger.

20. 20. The method of claim 19, comprising administering to the subject a dose of 10 mg per day of the compound of formula (I).

21. 20. The method of claim 19, comprising administering to the subject a dose of 7.5 mg per day of the compound of formula (I).

22. 20. The method of claim 19, comprising administering to the subject a dose of 5 mg per day of the compound of formula (I).

23. 20. The method of claim 19, comprising administering to the subject a dose of 2.5 mg per day of the compound of formula (I).

24. 20. The method of claim 19, comprising administering to the subject a dose of 2 mg per day of the compound of formula (I).

25. 20. The method of claim 19, comprising administering to the subject a dose of 1 mg per day of the compound of formula (I).

26. 26. The method of any one of claims 1-25, wherein said administering comprises administering a first dose of a fast-twitch myosin (type II) inhibitor and one or more subsequent doses of said fast-twitch myosin (type II) inhibitor, optionally wherein said one or more subsequent doses are higher than said first dose.

27. 27. The method of any one of claims 1-26, wherein said administering maintains or increases the North Star Ambulatory Assessment (NSAA) score of the subject compared to the NSAA score before treatment.

28. 28. The method of any one of claims 1-27, wherein said administering maintains or improves one or more of the following functional measures in the subject: maximum elbow flexion strength compared to maximum elbow flexion strength before treatment; maximum knee extension strength compared to maximum knee extension strength before treatment; 10 meter walk / run speed compared to 10 meter walk / run speed before treatment; 100 meter timed test speed compared to 100 meter timed test speed before treatment; 4 step stair climbing timed speed compared to 4 step stair climbing timed speed before treatment; and maximum grip strength compared to maximum grip strength value before treatment.

29. 29. The method of any one of claims 1-28, wherein said administering maintains or reduces the creatine kinase activity level in the subject by 10% or more compared to the creatine kinase activity level before treatment.

30. 29. The method of any one of claims 1-28, wherein said administering maintains or increases the creatine kinase activity level in the subject by 10% or more compared to the creatine kinase activity level before treatment.

31. 31. The method of any one of claims 1-30, wherein said administering maintains the subject's dual-energy x-ray absorptiometry (DXA) lean mass % or increases the subject's DXA lean mass % by 5% or more compared to pre-treatment DXA lean mass %.

32. 32. The method of any one of claims 1-31, wherein said administering maintains or reduces the subject's fast skeletal troponin I (TNNI2) concentration by 10% or more compared to the TNNI2 concentration before treatment, and optionally the TNNI2 concentration is estimated from SOMAscan levels.

33. 33. The method of any one of claims 1-32, wherein said administering reduces the protein concentration of one or more muscle injury biomarkers in said subject by 10% or more compared to pre-treatment protein concentration of said one or more muscle injury biomarkers.

34. 34. The method of claim 33, wherein the one or more muscle injury biomarkers comprise ACTN2, MYOM2, MYBPC1, PDLIM3, MYL3, TNNI2, MYL11, CKB, CKM, APOBEC2, ENO3, CA3, KLHL41, ADSS1, CAPN3, HSPB6, TNNT2, MYBPC2, GPD1(a), MUSTN1, FBP2, DUSP29, MYBPH, GPD1(b), THAP4, CHCD10, or a combination thereof.

35. 35. The method of any one of claims 1-34, wherein said administering reduces the protein concentration of one or more pro-inflammatory proteins in said subject by 10% or more compared to the pre-treatment protein concentration of said one or more pro-inflammatory proteins.

36. 36. The method of claim 35, wherein the one or more pro-inflammatory proteins comprise CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof.

37. 35. The method of any one of claims 1-34, wherein said administering increases the protein concentration of the one or more anti-inflammatory proteins in the subject by 10% or more compared to the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

38. 38. The method of claim 37, wherein the one or more anti-inflammatory proteins comprise IL10, IL13, IL22, IL37, IL4, or a combination thereof.

39. 39. The method of any preceding claim, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy and McArdle's disease.

40. 40. The method of claim 39, wherein the neuromuscular condition is Becker muscular dystrophy.

41. 40. The method of claim 39, wherein the neuromuscular condition is Duchenne muscular dystrophy.

42. 42. The method of any one of claims 1-41, wherein said administering comprises administering the compound of formula (I) for 2 months or more, 3 months or more, 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

43. 43. The method of claim 42, wherein said administering comprises administering the compound of formula (I) for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

44. 43. The method of claim 42, wherein said administering comprises administering said compound of formula (I) for six months or longer.

45. Prior to said administering, said method further comprises administering to a subject a composition comprising at least one of the following features (a) through (g): (a) creatine kinase activity levels of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) a TNNI2 concentration of 20 ng / mL or higher, 30 ng / mL or higher, or 40 ng / mL or higher; (c) dystrophin gene mutation; (d) pretreatment NSAA annual change of −0.1 to −10 or −0.5 to −3; (e) a pre-treatment serum creatinine of 1 mg / dL or less, 0.8 mg / dL or less, 0.6 mg / dL or less; (f) DXA % lean mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score of 1.5 or higher, or 2 or higher, associated with muscular dystrophy 45. The method of any one of claims 1 to 44, further comprising selecting a subject for treatment having one or more of:

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