Pyridazinone compositions for treatment of neuromuscular conditions

Abstract

Neuromuscular diseases, such as Duchenne Muscular Dystrophy (DMD), Bainite Muscular Dystrophy (BMD), Limb-banding Muscular Dystrophy (LGMD), and MecARDL Disease, are treated with compositions comprising substituted pyridazinone compounds.

Description

[0001] Cross-reference

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 63 / 375,180, filed on September 9, 2022, U.S. Provisional Application Serial No. 63 / 375,366, filed on September 12, 2022, U.S. Provisional Application Serial No. 63 / 510,348, filed on June 26, 2023, and U.S. Provisional Application Serial No. 63 / 580,334, filed on September 1, 2023, the entire contents of which are hereby incorporated herein by reference. BACKGROUND OF THE INVENTION

[0003] Skeletal muscle is the largest organ system in the human body and has two main functions. The first is force generation for muscle contraction, movement, and posture maintenance; the second is glucose, fatty acid, and amino acid metabolism. Skeletal muscle contraction during daily activities and exercise is naturally associated with muscle stress, breakdown, and remodeling, which are crucial for muscle adaptation. In individuals with neuromuscular conditions such as Duchenne Muscular Dystrophy (DMD), Becker Muscular Dystrophy (BMD), Limb-girdle muscular dystrophies (LGMD), and McArdle's disease, muscle contraction leads to successive rounds of amplified muscle breakdown that are difficult for the body to repair. Eventually, as the patient ages, pathophysiological processes occur, resulting in excessive inflammation, fibrosis, and accumulation of fat deposits in the muscle, heralding a sharp decline in physical function and leading to death. One potential treatment for such neuromuscular conditions is the administration of compounds that reduce skeletal muscle contraction. However, a decrease in skeletal muscle contractility can also lead to muscle weakness and a similar reduction in the patient's physical ability. There remains a need for treatments that reduce muscle breakdown in patients with neuromuscular conditions while maintaining or improving the patient's muscle strength. SUMMARY OF THE INVENTION

[0004] Described herein is a method of treating a neuromuscular condition, which comprises administering to a subject in need thereof a compound of formula (I) at a dose of 1 mg to 40 mg per day:

[0005] In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 15 mg to 40 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 15 mg, 20 mg, 25 mg or 30 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 20 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 1 mg to 20 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) 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 method comprises administering to the subject a dose of the compound of formula (I) of 1 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 2 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 2.5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 7.5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 10 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 15 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 20 mg per day. In some embodiments, the subject is over 18 years old. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 10 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 15 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) 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 dose of the compound of formula (I) of 10 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 7.5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 2.5 mg per day. In some embodiments, the method comprises administering to the subject a dose of the compound of formula (I) of 2 mg per day.In some embodiments, the method comprises administering to the subject a dose of 1 mg / day of the compound of formula (I). In some embodiments, the method comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose. In some embodiments, the method comprises maintaining or increasing the subject's NSAA score relative to the pre-treatment North Star Ambulatory Assessment (NSAA) score. In some embodiments, the method maintains or improves one or more of the following functional metrics of the subject: maximum elbow flexion strength relative to pre-treatment maximum elbow flexion strength; maximum knee extension strength relative to pre-treatment maximum knee extension strength; 10-meter walk / run speed relative to pre-treatment 10-meter walk / run speed; 100-meter timed test speed relative to pre-treatment 100-meter timed test speed; 4-step climb time speed relative to pre-treatment 4-step climb time speed; and maximum grip strength relative to pre-treatment maximum grip strength value. In some embodiments, relative to the pre-treatment creatine kinase activity level, the method maintains the creatine kinase activity level or reduces the creatine kinase activity level of the subject by 10% or more. In some embodiments, relative to the pre-treatment creatine kinase activity level, the method maintains the creatine kinase activity level or increases the creatine kinase activity level of the subject by 10% or more. In some embodiments, relative to the pre-treatment dual-energy x-ray absorptiometry (DXA) % lean body mass, the method maintains the subject's DXA % lean body mass or increases the subject's DXA % lean body mass by 5% or more. In some embodiments, relative to the pre-treatment skeletal muscle fast troponin I (TNNI2) concentration, the method maintains the subject's TNNI2 concentration or reduces the subject's TNNI2 concentration by 10% or more, and optionally wherein the TNNI2 concentration is inferred from the SOMAscan level. In some embodiments, relative to the pre-treatment protein concentration of one or more muscle injury biomarkers, the administration reduces the protein concentration of the one or more muscle injury biomarkers in the subject by 10% or more. 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, relative to the pre-treatment protein concentration of one or more pro-inflammatory proteins, the administration reduces the protein concentration of the one or more pro-inflammatory proteins in the subject by 10% or more. 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, relative to the pre-treatment protein concentration of one or more anti-inflammatory proteins, the administration increases the protein concentration of the one or more anti-inflammatory proteins in the subject by 10% or more. 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 neuromuscular condition is Becker muscular dystrophy. In some embodiments, the neuromuscular condition is Duchenne muscular dystrophy. In some embodiments, the administration comprises administering the compound of formula (I) for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer. In some embodiments, the administration comprises administering the compound of formula (I) for 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer. In some embodiments, the administration comprises administering the compound of formula (I) for 6 months or longer. In some embodiments, prior to the administration, the method further comprises selecting a subject having one or more of the characteristics (a)-(g) for treatment: (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 greater, 30 ng / mL or greater, or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) a pre-treatment annual change in NSAA of -0.1 to -10, or -0.5 to -3; (e) a pre-treatment serum creatinine of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) a DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0006] Disclosed herein is a method of treating a neuromuscular condition, comprising administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain or increase the North Star Ambulatory Assessment (NSAA) score of the subject relative to the pre-treatment NSAA score.

[0007] The present disclosure describes a method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) fast myosin (type II) inhibitor, the amount being sufficient to maintain or improve one or more of the following functional metrics of the subject: 100-meter timed test speed relative to the 100-meter timed test speed before treatment; 4-step climb time speed relative to the 4-step climb time speed before treatment; and maximum grip strength relative to the maximum grip strength value before treatment.

[0008] The present disclosure describes a method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) fast myosin (type II) inhibitor, the amount being sufficient to maintain the creatine kinase activity level or reduce the creatine kinase activity level of the subject by 10% or more relative to the creatine kinase activity level before treatment.

[0009] The present disclosure describes a method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) fast myosin (type II) inhibitor, the amount being sufficient to maintain the serum creatinine level or increase the serum creatinine level of the subject by 10% or more relative to the serum creatinine level before treatment.

[0010] The present disclosure describes a method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) fast myosin (type II) inhibitor, the amount being sufficient to maintain the DXA % lean body mass of the subject or increase the DXA % lean body mass of the subject by 5% or more relative to the DXA % lean body mass before treatment.

[0011] The present disclosure describes a method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) fast myosin (type II) inhibitor, the amount being sufficient to maintain the TNNI2 concentration of the subject or reduce the TNNI2 concentration of the subject by 10% or more relative to the skeletal muscle fast troponin I (TNNI2) concentration before treatment.

[0012] This text describes a method for inhibiting fast myosin (type II) in a subject in need thereof, which comprises selecting a subject having one or more of the 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 greater, 30 ng / mL or greater, or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA before treatment of -0.1 to -10, -0.5 to -3, or -1 to -4; (e) serum creatinine before treatment of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) DXA % lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater; and administering a fast myosin (type II) inhibitor to the subject.

[0013] This text describes a unit dose for oral administration, which comprises a compound of formula (I) in an amount of about 1 mg to about 20 mg and a pharmaceutically acceptable excipient.

[0014] This text describes a kit, which comprises an oral pharmaceutical composition and instructions for administering to a subject in need thereof, wherein the oral pharmaceutical composition comprises a compound of formula (I): and a pharmaceutically acceptable excipient.

[0015] This text describes a method for treating a neuromuscular condition, which comprises administering to a subject an amount of a compound of formula (I): said amount being sufficient to maintain the AUC of the subject 24 at 1000 ng / mL or higher.

[0016] This text describes a method for treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor, said amount being sufficient to maintain the myoglobin level or reduce the myoglobin level of the subject by 10% or more relative to the pre-treatment myoglobin level.

[0017] This text describes a method for alleviating pain associated with a neuromuscular condition, which comprises administering a fast myosin (type II) inhibitor to a subject in need thereof.

[0018] Incorporated by reference

[0019] All publications, patents, and patent applications mentioned in this specification are incorporated herein by reference to the extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. The patent publications WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, WO2021231615, and the PCT application PCT / US2023 / 021739 are incorporated herein by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] 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 invention will be obtained from the following detailed description that sets forth illustrative embodiments, which utilize the principles of the invention, and the accompanying drawings (also referred to herein as "figures"), in which:

[0021] Figure 1 Depicts the study results of BMD patients, showing disease progression as measured by NSAA.

[0022] Figure 2 Depicts the NSAA scores of BMD patients four months after administration of Compound 1.

[0023] Figure 3 Depicts the baseline characteristics of BMD patients compared to age normative values.

[0024] Figure 4 Depicts the plasma levels of Compound 1 and the step counts of BMD patients four months after dosing. For the 20 mg, 10 mg, and 15 mg doses, the mean AUC of the Compound 1 plasma PK levels were 2,293 ng / mL, 1,454 ng / mL, and 2,299 ng / mL, respectively.

[0025] Figure 5 Depicts the measurements of creatine kinase activity and skeletal muscle fast troponin I (TNNI2) levels in BMD patients after administration of Compound 1 compared to baseline measurements.

[0026] Figure 6 Depicts the NSAA scores of patients two and four months after initial administration of Compound 1 compared to baseline measurements.

[0027] Figure 7 Depicts the increase in functional measures (such as walking / running speed, stair climbing speed, and grip strength) in BMD patients two and four months after initial administration of Compound 1 compared to baseline measurements.

[0028] Figure 8 Depicts the reduction of self-reported pain scores in BMD patients at two and four months after the initial administration of Compound 1.

[0029] Figure 9A 、 Figure 9B and Figure 9C Depicts the North Star Ambulatory Assessment Scale.

[0030] Figures 10A - 10G Depicts the changes in several biomarkers after treatment with Compound 1. **p < 0.01; ***p < 0.001. Figure 10A Depicts the mean creatine kinase (CK) levels in different dosing regimens. All P < 0.05 except at month 3. Figure 10B Depicts the mean skeletal muscle fast troponin I (TNNI2) levels in different dosing regimens. All P < 0.05 except at month 4. Figure 10C Depicts the mean myoglobin levels in different dosing regimens. All P < 0.05 except at months 2, 3, 4, 6, or 10. Figure 10D Depicts the change in CK activity for each patient from the 12-month treatment group. p = 0.0087. Figure 10E Depicts the mean CK levels between baseline and the 12-month treatment group. p = 0.0012. The specified percentage difference is relative to the mean baseline; the bar graph shows the mean ± SEM. Figure 10F Depicts the change in TNNI2 activity for each patient in the 12-month treatment group. p = 0.022. TNNI2 data were inferred from SOMAscan. Figure 10G Depicts the mean TNNI2 levels between baseline and the 12-month treatment group. p < 0.0001. The specified percentage difference is relative to the mean baseline; the figure shows the mean ± SEM. TNNI2 data were inferred from SOMAscan.

[0031] Figures 11A - 11D Depicts the changes in the NSAA scores of Compound 1 and its plasma concentration and their correlation in different dosing regimens. Figure 11A Depicts the changes in the NSAA scores in different dosing regimens. The dashed line shows the mean ± 95% CI. The annotated lines showing a downward trend reflect the natural history data based on Luca Bello et al., MDA (2022) and van de Velde NM et al., Neurology (2021). Figure 11B Depicts the plasma concentration of Compound 1 in different dosing regimens. Figure 11C Depicts the NSAA response of individual patients in the 12-month group. The dashed line represents the expected reduction in the natural course of untreated patients. Figure 11D Depicts the corresponding plasma concentration of Compound 1 from the 12-month group.

[0032] Figures 12A - 12F Depicts the functional evaluation after treatment with Compound 1. Figure 12A Depicts the average 10m speed in different dosing regimens. All p-values were NS, with a 7% decrease at 12 months. Last observation carried forward method (all N = 12 except for 2 missing values at months 4 and 8). Figure 12B Depicts the average 100m speed in different dosing regimens. All p-values were NS, with a 0% change at 12 months. Last observation carried forward method (all N = 12 except for 2 missing values at months 4 and 8). Figure 12C Depicts the average 4-step climb time speed in different dosing regimens. All p-values were NS except for a p-value of 0.016 at 12 months, with a 13% decrease at 12 months. Figure 12D Depicts the average maximum grip strength in different dosing regimens. p-values < 0.05 at months 4, 6, and 8, and NS at 12 months. 8% decrease at 12 months. Figure 12E Depicts the average maximum knee extension strength in different dosing regimens. P-values < 0.05 at months 4, 8, and 12. 25% decrease at 12 months. Figure 12F Depicts the maximum elbow flexion strength in different dosing regimens. All p-values were NS and no change at 12 months.

[0033] Figure 13 Depicts the self-reported pain score after treatment with Compound 1. The figure shows mean ± SEM.

[0034] Figures 14A - 14H Illustrates the characterization of the short-term and long-term proteomic responses of skeletal muscle fast myosin (e.g., Compound 1) in Becker muscular dystrophy (BMD). Detailed Description

[0035] While the preferred embodiments of the present invention have been shown and described herein, it will be apparent to those skilled in the art that these embodiments are provided by way of example only. Many variations, changes, and substitutions will occur to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the present invention described herein may be employed in practicing the present invention. It is intended that the scope of the present invention be defined by the appended claims, which will cover methods and structures within the scope of these claims and their equivalents.

[0036] In some aspects, described herein is a method for treating a neuromuscular condition, which comprises administering to a subject in need thereof a dose of a compound of formula (I) of 1 mg to 40 mg per day: In some embodiments, the method comprises administering to a subject a dose of a compound of formula (I) of 15 mg to 40 mg per day. In some embodiments, the method comprises administering to a subject a dose of a compound of formula (I) 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 method comprises administering to a subject a dose of a compound of formula (I) of 10 mg, 15 mg or 20 mg per day when the subject is over 18 years of age. In some embodiments, the method comprises administering to a subject a dose of a compound of formula (I) of 10 mg, 7.5 mg, 5 mg, 2.5 mg, 2 mg or 1 mg per day when the subject is less than 18 years of age. In some embodiments, the administration comprises administering a first dose of the fast myosin (type II) fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose. In some embodiments, the administration maintains the North Star Ambulatory Assessment (NSAA) score of the subject or increases the NSAA score of the subject relative to the pre-treatment NSAA score. In some embodiments, the administration maintains or improves one or more of the following functional metrics of the subject: maximum elbow flexion strength relative to pre-treatment maximum elbow flexion strength; maximum knee extension strength relative to pre-treatment maximum knee extension strength; 10-meter walking / running speed relative to pre-treatment 10-meter walking / running speed; 100-meter timed test speed relative to pre-treatment 100-meter timed test speed; 4-step climb time speed relative to pre-treatment 4-step climb time speed; and maximum grip strength relative to pre-treatment maximum grip strength value. In some embodiments, the administration maintains the creatine kinase activity level or reduces the creatine kinase activity level of the subject by 10% or more relative to the pre-treatment creatine kinase activity level. In some embodiments, the administration maintains the creatine kinase activity level or increases the creatine kinase activity level of the subject by 10% or more relative to the pre-treatment creatine kinase activity level. In some embodiments, the administration maintains the DXA % lean body mass of the subject or increases the DXA % lean body mass of the subject by 5% or more relative to the pre-treatment dual energy x-ray absorptiometry (DXA) % lean body mass. In some embodiments, the administration maintains the TNNI2 concentration of the subject or reduces the TNNI2 concentration of the subject by 10% or more relative to the pre-treatment skeletal muscle fast troponin I (TNNI2) concentration, and optionally wherein the TNNI2 concentration is inferred from the SOMAscan level. In some embodiments, the administration reduces the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more relative to the pre-treatment protein concentration of one or more muscle injury biomarkers.In some embodiments, 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 administration reduces the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more relative to the pre-treatment protein concentration of the one or more pro-inflammatory proteins. In some embodiments, one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In some embodiments, the administration increases the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more relative to the pre-treatment protein concentration of the one or more anti-inflammatory proteins. In some embodiments, 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 administration comprises administering the compound of formula (I) for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer. In some embodiments, prior to the administration, the method further comprises selecting a subject having one or more of the characteristics (a)-(g) for treatment: (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 greater, 30 ng / mL or greater, or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) a pre-treatment annual change in NSAA of -0.1 to -10, or -0.5 to -3; (e) a pre-treatment serum creatinine of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0037] In some aspects, the present disclosure provides methods and compositions for treating neuromuscular conditions by selectively inhibiting fast-twitch skeletal muscle myosin. In particular, the methods of the present disclosure can be used to treat DMD, BMD, LGMD, McArdle's disease, and other neuromuscular conditions. In some embodiments, the methods described herein include compounds or salts previously described in publications WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615.

[0038] Skeletal muscle is mainly composed of two types of fibers, slow-twitch muscle fibers (i.e., type I) and fast-twitch muscle fibers (i.e., type II). In each muscle, the two types of fibers are arranged in a mosaic configuration, and the fiber type composition varies in different muscles and at different points of growth and development. Slow-twitch muscle fibers have excellent aerobic energy production capabilities. Slow-twitch muscle fibers have a low contraction rate but high fatigue tolerance. Slow-twitch muscle fibers generally have a higher concentration of mitochondria and myoglobin than fast-twitch fibers and are surrounded by more capillaries compared to fast-twitch fibers. Due to the lower myosin ATPase activity, slow-twitch fibers contract more slowly and produce less energy compared to fast-twitch fibers, but they are able to maintain contraction function for a longer time, such as in steady-state, postural control, and endurance exercises.

[0039] Human fast-twitch muscle fibers are further divided into two main fiber types (type IIa, type IIx / d) based on the specific skeletal muscle fast myosin they express. A third type of fast-twitch fiber (type IIb) is present in other mammals but is rarely found in human muscle. Fast-twitch muscle fibers have excellent anaerobic energy production capabilities and are able to generate a large amount of tension in a short time. Generally, compared to slow-twitch muscle fibers, fast-twitch muscle fibers have lower concentrations of mitochondria, myoglobin, and capillaries and thus fatigue more quickly. Fast-twitch muscles generate the faster forces required for power and resistance activities.

[0040] The ratio of type I and type II can vary among different individuals. For example, each muscle fiber type can account for nearly 50% in non-athletic individuals. Power athletes may have a higher proportion of fast-twitch fibers. For example, 70%-75% of type II fibers are present in sprinters. Endurance athletes may have a higher proportion of slow-twitch fibers. For example, the proportion of slow-twitch fibers in long-distance runners is 70%-80%. The ratio of type I and type II fibers can also vary according to the age of the individual. The proportion of type II fibers (especially type IIx) can decrease with increasing age of the individual, resulting in loss of lean muscle mass.

[0041] The contractile action of skeletal muscle results in muscle damage in subjects with neuromuscular diseases (e.g., DMD), and this damage appears to be more prevalent in fast-twitch muscle fibers. It has been observed that in a murine model of dystrophy, the acute force decline after strain in type II fibers, which are predominantly fast-twitch, is relatively greater than in type I fibers, which are predominantly slow-twitch. It has also been demonstrated in a murine dystrophy model that the degree of acute force decline and histological damage is proportional to the peak force development during strain. Damage caused by overcontraction occurs prior to the inflammation and irreversible fibrosis characteristic of late-stage DMD pathology.

[0042] Skeletal muscle myosin inhibitors that are not selective for type II fibers can lead to overinhibition of skeletal muscle contraction (including respiratory function) and unwanted inhibition of cardiac activity because the heart shares multiple structural components (such as type I myosin) with type I skeletal muscle fibers. Without wishing to be bound by a theory of a particular mechanism, 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's 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.

[0043] When healthy muscle is subjected to excessive, unaccustomed exercise, it can result in soreness and a persistent reduction in strength and range of motion. Proteins, including creatine kinase (CK), lactate dehydrogenase, and myoglobin, also leak from damaged muscle fibers into the circulation. These biomarkers are not specific to fast-twitch or slow-twitch muscle fibers and thus do not provide detailed information about differences in the response of damaged fibers. Troponin I (TNNI) is a component of the troponin complex, which controls the initiation of muscle contraction through calcium. The difference is that each type of striated muscle has a different isoform: TNNI1 in slow-twitch skeletal muscle, TNNI2 in fast-twitch skeletal muscle, and TNNI3 in cardiac muscle. It has been demonstrated using a selective enzyme-linked immunosorbent assay (ELISA) that TNNI2, but not TNNI1, is elevated in the circulation after damaging exercise, even under extreme conditions.

[0044] DMD and BMD are caused by the absence (DMD) or truncation (BMD) of dystrophin. Dystrophin provides a structural link between the actin cytoskeleton and the basement membrane through the dystrophin-glycoprotein complex. When dystrophin is absent or truncated, muscle contraction leads to increased muscle stress and damage during normal use. Although DMD muscles are far more sensitive to injury than BMD or healthy muscles, fast-twitch muscle fibers still appear to be more sensitive than slow-twitch muscle fibers, with histological evidence of fast-twitch muscle fiber 7 disruption and early loss of type IIx fibers in young DMD patients. In some embodiments, the present disclosure provides selective inhibitors of fast-twitch skeletal muscle myosin as a treatment option for DMD, BMD, McArdle's disease, or limb-girdle muscular dystrophy.

[0045] Definitions

[0046] 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.

[0047] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise.

[0048] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, typically by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, intratracheal, subcutaneous, subepidermal, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0049] The phrase "pharmaceutically acceptable" is used herein to mean those compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications and are commensurate with a reasonable benefit / risk ratio.

[0050] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" 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 injurious 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 carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) diols, 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) ethanol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0051] As used herein, "treatment" or "treating" refers to a method for obtaining a beneficial or desired result (including, but not limited to, therapeutic and / or prophylactic benefits) with respect to a disease, disorder, or medical condition. Therapeutic benefits can include, for example, eradication or amelioration of the underlying disorder being treated. Additionally, therapeutic benefits can include, for example, eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder, such that an improvement is observed in the subject, even though the subject may still be afflicted with the underlying disorder. In certain embodiments, for prophylactic benefits, the composition is administered to a subject at risk of developing a particular disease or reporting one or more physiological symptoms of a disease, even if a diagnosis of the disease has not yet been made. Treatment by administration of the compounds described herein does not require the involvement of a medical professional.

[0052] 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), also known as fast myosin (type II), and are referred to as fast myosin (type II) inhibitors. In certain embodiments, the compounds and salts for use in the methods and compositions of the present disclosure are described in any one of WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615. In certain embodiments, the compounds for use in the methods described herein are depicted as compounds of formula (I):

[0053] Also referred to herein as Compound 1.

[0054] Therapeutic applications

[0055] In some aspects, methods for treating neuromuscular conditions are described herein, which comprise administering to a subject in need thereof a dose of a compound of formula (I) of 1 mg to 40 mg per day: In some embodiments, administration to a subject comprises a dose of the compound of formula (I) from 15 mg to 40 mg per day. In some embodiments, administration to a subject comprises a dose of the compound of formula (I) of 15 mg, 20 mg, 25 mg or 30 mg per day. In some embodiments, the dose of the compound of formula (I) comprises from about 1 mg per day to about 30 mg per day. In some embodiments, the dose of the compound of formula (I) comprises from about 1 mg per day to about 2 mg per day, from about 1 mg per day to about 2.5 mg per day, from about 1 mg per day to about 3 mg per day, from about 1 mg per day to about 5 mg per day, from about 1 mg per day to about 7.5 mg per day, from about 1 mg per day to about 10 mg per day, from about 1 mg per day to about 12.5 mg per day, from about 1 mg per day to about 15 mg per day, from about 1 mg per day to about 20 mg per day, from about 1 mg per day to about 25 mg per day, from about 1 mg per day to about 30 mg per day, from about 2 mg per day to about 2.5 mg per day, from about 2 mg per day to about 3 mg per day, from about 2 mg per day to about 5 mg per day, from about 2 mg per day to about 7.5 mg per day, from about 2 mg per day to about 10 mg per day, from about 2 mg per day to about 12.5 mg per day, from about 2 mg per day to about 15 mg per day, from about 2 mg per day to about 20 mg per day, from about 2 mg per day to about 25 mg per day, from about 2 mg per day to about 30 mg per day, from about 2.5 mg per day to about 3 mg per day, from about 2.5 mg per day to about 5 mg per day, from about 2.5 mg per day to about 7.5 mg per day, from about 2.5 mg per day to about 10 mg per day, from about 2.5 mg per day to about 12.5 mg per day, from about 2.5 mg per day to about 15 mg per day, from about 2.5 mg per day to about 20 mg per day, from about 2.5 mg per day to about 25 mg per day, from about 2.5 mg per day to about 30 mg per day, from about 3 mg per day to about 5 mg per day, from about 3 mg per day to about 7.5 mg per day, from about 3 mg per day to about 10 mg per day, from about 3 mg per day to about 12.5 mg per day, from about 3 mg per day to about 15 mg per day, from about 3 mg per day to about 20 mg per day, from about 3 mg per day to about 25 mg per day, from about 3 mg per day to about 30 mg per day, from about 5 mg per day to about 7.5 mg per day, from about 5 mg per day to about 10 mg per day, from about 5 mg per day to about 12.5 mg per day, from about 5 mg per day to about 15 mg per day, from about 5 mg per day to about 20 mg per day, from about 5 mg per day to about 25 mg per day, from about 5 mg per day to about 30 mg per day, from about 7.5 mg per day to about 10 mg per day, from about 7.5 mg per day to about 12.5 mg per day, from about 7.5 mg per day to about 15 mg per day, from about 7.5 mg per day to about 20 mg per day, from about 7.5 mg per day to about 25 mg per day, from about 7.5 mg per day to about 30 mg per day, from about 10 mg per day to about 12.5 mg / day, from about 10 mg / day to about 15 mg / day, from about 10 mg / day to about 20 mg / day, from about 10 mg / day to about 25 mg / day, from about 10 mg / day to about 30 mg / day, from about 12.5 mg / day to about 15 mg / day, from about 12.5 mg / day to about 20 mg / day, from about 12.5 mg / day to about 25 mg / day, from about 12.5 mg / day to about 30 mg / day, from about 15 mg / day to about 20 mg / day, from about 15 mg / day to about 25 mg / day, from about 15 mg / day to about 30 mg / day, from about 20 mg / day to about 25 mg / day, from about 20 mg / day to about 30 mg / day, or from about 25 mg / day to about 30 mg / day. In some embodiments, the dose of the compound of formula (I) comprises about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day. In some embodiments, the dose of the compound of formula (I) comprises at least about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, or about 25 mg / day. In some embodiments, the dose of the compound of formula (I) comprises at most about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day.

[0056] In some embodiments, the dosage of the compound of formula (I) comprises at least about 1 mg / day to about 30 mg / day. In some embodiments, the dosage of the compound of formula (I) comprises at least about 1 mg / day to about 2 mg / day, about 1 mg / day to about 2.5 mg / day, about 1 mg / day to about 3 mg / day, about 1 mg / day to about 5 mg / day, about 1 mg / day to about 7.5 mg / day, about 1 mg / day to about 10 mg / day, about 1 mg / day to about 12.5 mg / day, about 1 mg / day to about 15 mg / day, about 1 mg / day to about 20 mg / day, about 1 mg / day to about 25 mg / day, about 1 mg / day to about 30 mg / day, about 2 mg / day to about 2.5 mg / day, about 2 mg / day to about 3 mg / day, about 2 mg / day to about 5 mg / day, about 2 mg / day to about 7.5 mg / day, about 2 mg / day to about 10 mg / day, about 2 mg / day to about 12.5 mg / day, about 2 mg / day to about 15 mg / day, about 2 mg / day to about 20 mg / day, about 2 mg / day to about 25 mg / day, about 2 mg / day to about 30 mg / day, about 2.5 mg / day to about 3 mg / day, about 2.5 mg / day to about 5 mg / day, about 2.5 mg / day to about 7.5 mg / day, about 2.5 mg / day to about 10 mg / day, about 2.5 mg / day to about 12.5 mg / day, about 2.5 mg / day to about 15 mg / day, about 2.5 mg / day to about 20 mg / day, about 2.5 mg / day to about 25 mg / day, about 2.5 mg / day to about 30 mg / day, about 3 mg / day to about 5 mg / day, about 3 mg / day to about 7.5 mg / day, about 3 mg / day to about 10 mg / day, about 3 mg / day to about 12.5 mg / day, about 3 mg / day to about 15 mg / day, about 3 mg / day to about 20 mg / day, about 3 mg / day to about 25 mg / day, about 3 mg / day to about 30 mg / day, about 5 mg / day to about 7.5 mg / day, about 5 mg / day to about 10 mg / day, about 5 mg / day to about 12.5 mg / day, about 5 mg / day to about 15 mg / day, about 5 mg / day to about 20 mg / day, about 5 mg / day to about 25 mg / day, about 5 mg / day to about 30 mg / day, about 7.5 mg / day to about 10 mg / day, about 7.5 mg / day to about 12.5 mg / day, about 7.5 mg / day to about 15 mg / day, about 7.5 mg / day to about 20 mg / day, about 7.5 mg / day to about 25 mg / day, about 7.5 mg / day to about 30 mg / day, about 10 mg / day to about 12.5 mg / day, about 10 mg / day to about 15 mg / day, about 10 mg / day to about 20 mg / day, about 10 mg / day to about 25 mg / day, about 10 mg / day to about 30 mg / day, about 12.5 mg / day to about 15 mg / day, about 12.5 mg / day to about 20 mg / day, about 12.5 mg / day to about 25 mg / day, about 12.5 mg / day to about 30 mg / day, about 15 mg / day to about 20 mg / day, about 15 mg / day to about 25 mg / day, about 15 mg / day to about 30 mg / day, about 20 mg / day to about 25 mg / day, about 20 mg / day to about 30 mg / day, or about 25 mg / day to about 30 mg / day. In some embodiments, the dose of the compound of formula (I) comprises at least about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day. In some embodiments, the dose of the compound of formula (I) comprises at least at least about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, or about 25 mg / day. In some embodiments, the dose of the compound of formula (I) comprises at least at most about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day.

[0057] In some embodiments, the dosage of the compound of formula (I) comprises up to about 1 mg / day to about 30 mg / day. In some embodiments, the dosage of the compound of formula (I) comprises up to about 1 mg / day to about 2 mg / day, about 1 mg / day to about 2.5 mg / day, about 1 mg / day to about 3 mg / day, about 1 mg / day to about 5 mg / day, about 1 mg / day to about 7.5 mg / day, about 1 mg / day to about 10 mg / day, about 1 mg / day to about 12.5 mg / day, about 1 mg / day to about 15 mg / day, about 1 mg / day to about 20 mg / day, about 1 mg / day to about 25 mg / day, about 1 mg / day to about 30 mg / day, about 2 mg / day to about 2.5 mg / day, about 2 mg / day to about 3 mg / day, about 2 mg / day to about 5 mg / day, about 2 mg / day to about 7.5 mg / day, about 2 mg / day to about 10 mg / day, about 2 mg / day to about 12.5 mg / day, about 2 mg / day to about 15 mg / day, about 2 mg / day to about 20 mg / day, about 2 mg / day to about 25 mg / day, about 2 mg / day to about 30 mg / day, about 2.5 mg / day to about 3 mg / day, about 2.5 mg / day to about 5 mg / day, about 2.5 mg / day to about 7.5 mg / day, about 2.5 mg / day to about 10 mg / day, about 2.5 mg / day to about 12.5 mg / day, about 2.5 mg / day to about 15 mg / day, about 2.5 mg / day to about 20 mg / day, about 2.5 mg / day to about 25 mg / day, about 2.5 mg / day to about 30 mg / day, about 3 mg / day to about 5 mg / day, about 3 mg / day to about 7.5 mg / day, about 3 mg / day to about 10 mg / day, about 3 mg / day to about 12.5 mg / day, about 3 mg / day to about 15 mg / day, about 3 mg / day to about 20 mg / day, about 3 mg / day to about 25 mg / day, about 3 mg / day to about 30 mg / day, about 5 mg / day to about 7.5 mg / day, about 5 mg / day to about 10 mg / day, about 5 mg / day to about 12.5 mg / day, about 5 mg / day to about 15 mg / day, about 5 mg / day to about 20 mg / day, about 5 mg / day to about 25 mg / day, about 5 mg / day to about 30 mg / day, about 7.5 mg / day to about 10 mg / day, about 7.5 mg / day to about 12.5 mg / day, about 7.5 mg / day to about 15 mg / day, about 7.5 mg / day to about 20 mg / day, about 7.5 mg / day to about 25 mg / day, about 7.5 mg / day to about 30 mg / day, about 10 mg / day to about 12.5 mg / day, about 10 mg / day to about 15 mg / day, about 10 mg / day to about 20 mg / day, about 10 mg / day to about 25 mg / day, about 10 mg / day to about 30 mg / day, about 12.5 mg / day to about 15 mg / day, about 12.5 mg / day to about 20 mg / day, about 12.5 mg / day to about 25 mg / day, about 12.5 mg / day to about 30 mg / day, about 15 mg / day to about 20 mg / day, about 15 mg / day to about 25 mg / day, about 15 mg / day to about 30 mg / day, about 20 mg / day to about 25 mg / day, about 20 mg / day to about 30 mg / day, or about 25 mg / day to about 30 mg / day. In some embodiments, the dose of the compound of formula (I) includes up to about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day. In some embodiments, the dose of the compound of formula (I) includes at least up to about 1 mg / day, about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, or about 25 mg / day. In some embodiments, the dose of the compound of formula (I) includes at most up to about 2 mg / day, about 2.5 mg / day, about 3 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 20 mg / day, about 25 mg / day, or about 30 mg / day.

[0058] In some embodiments, the administration to the subject includes a dose of 20 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 1 mg to 20 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes 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 / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 1 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 2 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 2.5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 7.5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 10 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 15 mg / day of the compound of formula (I). In some embodiments, the administration to the subject includes a dose of 20 mg / day of the compound of formula (I).

[0059] In some embodiments, the subject being treated with the compound of formula (I) is over 18 years of age. In some embodiments, the administration to the subject comprises a dose of 10 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 15 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 20 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 10 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 7.5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 2.5 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 2 mg / day of the compound of formula (I). In some embodiments, the administration to the subject comprises a dose of 1 mg / day of the compound of formula (I). In some embodiments, the administration comprises administering a first dose of the fast myosin (type II) fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose. In some embodiments, the administration maintains the North Star Ambulatory Assessment (NSAA) score of the subject or increases the NSAA score of the subject relative to the pre-treatment NSAA score. In some embodiments, the administration maintains or improves one or more of the following functional metrics of the subject: maximum elbow flexion strength relative to pre-treatment maximum elbow flexion strength; maximum knee extension strength relative to pre-treatment maximum knee extension strength; 10-meter walk / run speed relative to pre-treatment 10-meter walk / run speed; 100-meter timed test speed relative to pre-treatment 100-meter timed test speed; 4-step climb time speed relative to pre-treatment 4-step climb time speed; and maximum grip strength relative to pre-treatment maximum grip strength value. In some embodiments, the administration maintains the creatine kinase activity level or reduces the creatine kinase activity level of the subject by 10% or more relative to the pre-treatment creatine kinase activity level. In some embodiments, the administration maintains the creatine kinase activity level or increases the creatine kinase activity level of the subject by 10% or more relative to the pre-treatment creatine kinase activity level. In some embodiments, the administration maintains the DXA % lean body mass of the subject or increases the DXA % lean body mass of the subject by 5% or more relative to the pre-treatment dual energy x-ray absorptiometry (DXA) % lean body mass. In some embodiments, the administration maintains the TNNI2 concentration of the subject or reduces the TNNI2 concentration of the subject by 10% or more relative to the pre-treatment skeletal muscle fast troponin I (TNNI2) concentration, and optionally wherein the TNNI2 concentration is inferred from the SOMAscan level.In some embodiments, relative to the pre-treatment protein concentration of one or more muscle injury biomarkers, the administration reduces the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more. In some embodiments, 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, relative to the pre-treatment protein concentration of one or more pro-inflammatory proteins, the administration reduces the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more. In some embodiments, one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In some embodiments, relative to the pre-treatment protein concentration of one or more anti-inflammatory proteins, the administration increases the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more. In some embodiments, 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 administration comprises administering a compound of formula (I) for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer. In some embodiments, prior to the administration, the method further comprises selecting a subject having one or more of the characteristics (a)-(g) for treatment: (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 greater, 30 ng / mL or greater, or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA prior to treatment of -0.1 to -10, or -0.5 to -3; (e) a serum creatinine of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower prior to treatment; (f) a DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0060] The methods described herein can be used to treat neuromuscular conditions. In some embodiments, the methods include administering a composition that comprises a fast myosin (type II) fast myosin (type II) inhibitor and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents, and / or thickening agents. In some embodiments, the fast myosin (type II) fast 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, the fast myosin (type II) fast myosin (type II) inhibitor is described in WO2020097266. In some embodiments, the muscle myosin inhibitor is depicted by formula (I): Also known as Compound 1.

[0061] The administration of Compound 1 or another fast skeletal myosin (type II) fast skeletal myosin (type II) inhibitor can be used to treat neuromuscular conditions and motor disorders. Examples of neuromuscular conditions include, but are not limited to, Duchenne muscular dystrophy, Becker muscular dystrophy, myotonic dystrophy type 1, myotonic dystrophy type 2, facioscapulohumeral muscular dystrophy, oculopharyngeal muscular dystrophy, limb-girdle muscular dystrophy, tendinitis, and carpal tunnel syndrome. Examples of motor disorders include, but are not limited to, muscle spasticity disorders, spasticity associated with multiple sclerosis, Parkinson's disease, Alzheimer's disease, or cerebral palsy, or an injury or traumatic event (such as a stroke, traumatic brain injury, spinal cord injury, hypoxia, meningitis, encephalitis, phenylketonuria, or amyotrophic lateral sclerosis). Also included are other conditions that may respond to the 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. In some embodiments, the neuromuscular conditions and motor disorders are selected from muscular dystrophies and myopathies. In some embodiments, muscular dystrophy is a disease that causes progressive weakness and loss of muscle mass, in which an abnormal gene (mutation) interferes with the production of proteins required to form healthy muscle. In some embodiments, muscular dystrophy is 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, congenital muscular dystrophy (CMD) is selected from Bethlem CMD, Fukuyama CMD, muscle-eye-brain disease (MEB), rigid spine syndrome, Ullrich CMD, and Walker-Warburg syndromes (WWS). In some embodiments, myopathy is a muscle disease not caused by a neurological disorder. Myopathy causes the muscles to become weak or shrink (atrophy). In some embodiments, myopathy is selected from congenital myopathy, distal myopathy, endocrine myopathy, inflammatory myopathy, metabolic myopathy, myofibrillar myopathy (MFM), scapuloperoneal myopathy, and cardiomyopathy. In some embodiments, congenital myopathy is selected from capmyopathies, centronuclear myopathies, congenital myopathy with fiber type disproportion, core myopathies, central core disease, multiminicore myopathies, myosin storage myopathy, myotubular myopathy, and nemaline myopathy.In some embodiments, distal myopathy is selected from GNE myopathy / Nonaka myopathy / hereditary inclusion body myopathy (HIBM), Laing distal myopathy, Markesbery-Griggs late-onset distal myopathy, Miyoshi myopathy, Udd myopathy / tibial muscular dystrophy, VCP myopathy / IBMPFD, vocal cord and pharyngeal distal myopathy, and Welander distal myopathy. In some embodiments, endocrine myopathy is selected from hyperthyroid myopathy and hypothyroid myopathy. In some embodiments, inflammatory myopathy is selected from dermatomyositis, inclusion body myositis, and polymyositis. In some embodiments, 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, cardiomyopathy is selected from endogenous cardiomyopathy and exogenous cardiomyopathy. In some embodiments, endogenous cardiomyopathy is selected from hereditary myopathy and acquired myopathy. In some embodiments, hereditary myopathy is selected from hypertrophic cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy (ARVC), LV non-compaction, ion channelopathy, dilated cardiomyopathy (DCM), and restrictive cardiomyopathy (RCM). In some embodiments, acquired myopathy is selected from stress cardiomyopathy, myocarditis, eosinophilic myocarditis, and ischemic cardiomyopathy. In some embodiments, exogenous cardiomyopathy is selected from metabolic cardiomyopathy, endocardial cardiomyopathy, endocrine cardiomyopathy, and cardiofacial myopathy. In some embodiments, metabolic cardiomyopathy is selected from Fabry disease and hemochromatosis. In some embodiments, endocardial cardiomyopathy is selected from endomyocardial fibrosis and hypereosinophilic syndrome. In some embodiments, endocrine cardiomyopathy is selected from diabetes, hyperthyroidism, and acromegaly. In some embodiments, cardiofacial myopathy is Noonan syndrome.

[0062] In some aspects, methods of treating neuromuscular conditions or movement disorders can 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 cardiac muscle contraction. In some embodiments, cardiac muscle contraction is inhibited by 20% or less. In some embodiments, cardiac muscle contraction is inhibited by 15% or less. In some embodiments, cardiac muscle contraction is inhibited by 10% or less. In some embodiments, cardiac muscle contraction is inhibited by 9% or less. In some embodiments, cardiac muscle contraction is inhibited by 8% or less. In some embodiments, cardiac muscle contraction is inhibited by 7% or less. In some embodiments, cardiac muscle contraction is inhibited by 6% or less. In some embodiments, cardiac muscle contraction is inhibited by 5% or less. In some embodiments, cardiac muscle contraction is inhibited by 4% or less. In some embodiments, cardiac muscle contraction is inhibited by 3% or less. In some embodiments, cardiac muscle contraction is inhibited by 2% or less. In some embodiments, cardiac muscle contraction is inhibited by 1% or less.

[0063] Before and after treatment with Compound 1 or a salt thereof, the activities of daily living (ADL) or habitual physical activities of the subject can be monitored. The ADL or habitual physical activities depend on the subject and can range from simple walking to extensive exercise, depending on the subject's capabilities and routine. The treatment options and dosages of the skeletal muscle contraction inhibitors discussed herein can be personalized for the subject such that the ADL and habitual physical activities remain unchanged.

[0064] In some aspects, methods of treating neuromuscular conditions or movement disorders 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% relative to the subject's pre-treatment skeletal muscle contraction ability. Compound 1 or a salt thereof can be administered in an amount that reduces skeletal muscle contraction by 5% to 45% relative to the subject's pre-treatment skeletal muscle contraction ability. 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% relative to the subject's pre-treatment skeletal muscle contraction ability. In certain embodiments, Compound 1 or a salt thereof can be administered in an amount that reduces skeletal muscle contraction by 1% to 50% relative to the subject's pre-treatment skeletal muscle contraction ability.

[0065] In some aspects, a method of treating a neuromuscular condition or a movement disorder may include administering Compound 1 or a salt thereof to inhibit type I skeletal muscle contraction. The dosage of the type I skeletal muscle contraction inhibitor may be administered in an amount relative to the amount required to reduce type I skeletal muscle contraction by 20%. The type I skeletal muscle contraction inhibitor may be administered in an amount less than the amount required to reduce type I skeletal muscle contraction by 20% relative to the pre-treatment type I skeletal muscle contraction ability of the subject. The type I skeletal muscle contraction inhibitor may be administered in an amount that reduces type I skeletal muscle contraction by 0.01% to 20% relative to the pre-treatment type I skeletal muscle contraction ability of the subject. 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% relative to the pre-treatment type I skeletal muscle contraction ability of the subject. In certain embodiments, the inhibitor may be administered in an amount that reduces type I skeletal muscle contraction by 0.01% to 20% relative to the pre-treatment type I skeletal muscle contraction ability of the subject.

[0066] In some aspects, a method of treating a neuromuscular condition or a movement disorder may include administering Compound 1 or a salt thereof to inhibit type II skeletal muscle contraction. The dosage of the type II skeletal muscle contraction inhibitor may be administered in an amount relative to the amount required to reduce type II skeletal muscle contraction by 90%. The type II skeletal muscle contraction inhibitor may be administered in an amount less than the amount required to reduce type II skeletal muscle contraction by 90% relative to the pre-treatment type II skeletal muscle contraction ability of the subject. The type II skeletal muscle contraction inhibitor may be administered in an amount that reduces type II skeletal muscle contraction by 5% to 75% relative to the pre-treatment type II skeletal muscle contraction ability of the subject. In some cases, the inhibitor may be administered in an amount that reduces type II 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%, less than 50%, less than 55%, less than 60%, less than 65%, less than 70%, less than 75%, less than 80%, less than 85%, or even less than 90% relative to the pre-treatment type II skeletal muscle contraction ability of the subject. In certain embodiments, the inhibitor may be administered in an amount that reduces type II skeletal muscle contraction by 1% to 50% relative to the pre-treatment type II skeletal muscle contraction ability of the subject.

[0067] In some aspects, a method of treating contraction-induced skeletal muscle fiber injury may include administering Compound 1 or a salt thereof to inhibit skeletal muscle contraction and / or skeletal muscle fast myosin (type II) fast myosin (type II). In certain embodiments, the inhibitor does not significantly inhibit cardiac muscle contraction.

[0068] In some aspects, a method of treating a metabolic myopathy (such as McArdle's syndrome) may include administering Compound 1 or a salt thereof.

[0069] In certain embodiments, contraction-induced damage in skeletal muscle fibers results from involuntary skeletal muscle contractions. Involuntary skeletal muscle contractions can be associated with neuromuscular disorders or spasticity-related disorders. In certain embodiments, contraction-induced damage in skeletal muscle fibers can result from voluntary skeletal muscle contractions, such as physical exercise.

[0070] In certain embodiments, administering Compound 1 or a salt thereof to a subject modulates one or more biomarkers associated with 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 (such as cardiac, slow skeletal muscle, and fast skeletal muscle TnI), and inflammatory markers (IL-1, IL-6, IL-4, TNF-α). Biomarkers can also include a measure of muscle inflammation (e.g., edema). Relative to the pre-treatment level of the biomarker, the level of the biomarker described herein can increase after administering an inhibitor. Alternatively, relative to the pre-treatment level of the biomarker, the level of the biomarker can decrease after administering an inhibitor. Modulation of one or more biomarkers with the inhibitors described herein can indicate treatment of a neuromuscular disorder such as those described herein.

[0071] In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more muscle damage biomarkers (e.g., Figure 14D one or more muscle damage proteins as shown). Examples of one or more muscle damage biomarkers can 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, administering Compound 1 or a salt thereof to a subject reduces one or more muscle damage biomarkers during short-term treatment (e.g., treating with Compound 1 for 1-2 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more muscle damage biomarkers during medium-term treatment (e.g., treating with Compound 1 for 3-4 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more muscle damage biomarkers during long-term treatment (e.g., treating with Compound 1 for 6-12 months).

[0072] In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins (e.g., Figure 14G or Figure 14H one or more pro-inflammatory proteins as shown in). Examples of one or more pro-inflammatory proteins can include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof. In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during short-term treatment (e.g., treating with Compound 1 for 1 - 2 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during medium-term treatment (e.g., treating with Compound 1 for 3 - 4 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject reduces one or more pro-inflammatory proteins during long-term treatment (e.g., treating with Compound 1 for 6 - 12 months).

[0073] In certain embodiments, administering Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins (e.g., Figure 14G or Figure 14H one or more anti-inflammatory proteins as shown in). Examples of one or more anti-inflammatory proteins can include IL10, IL13, IL22, IL37, IL4, or a combination thereof. In certain embodiments, administering Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during short-term treatment (e.g., treating with Compound 1 for 1 - 2 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during medium-term treatment (e.g., treating with Compound 1 for 3 - 4 months). In certain embodiments, administering Compound 1 or a salt thereof to a subject increases one or more anti-inflammatory proteins during long-term treatment (e.g., treating with Compound 1 for 6 - 12 months).

[0074] Compared to when the subject is inactive (e.g., sleeping), the CK level in the subject increases during activity, and thus CK is a potential metric for assessing skeletal muscle breakdown caused by skeletal muscle contraction. In certain embodiments, Compound 1 or a salt thereof can be administered to the subject before mild, moderate, or vigorous activity to reduce or prevent skeletal muscle breakdown caused by the activity. Moderate to vigorous activity can depend on the subject's ability and can include physical exercise that increases the heart rate by at least 20% or more (such as about 50% or more) relative to the subject's resting heart rate. Examples of moderate to vigorous activity include walking, running, weightlifting, cycling, swimming, hiking, etc.

[0075] In some embodiments, a subject is administered an amount of a fast myosin (type II) inhibitor, such as Compound 1, that is sufficient to maintain the creatine kinase activity level or reduce the creatine kinase activity level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more relative to the pre-treatment creatine kinase activity level. In some embodiments, the creatine kinase activity level of the subject is evaluated after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the evaluation is performed for 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, 18 months or longer, or 24 months or longer. In some embodiments, the creatine kinase activity level of the subject is evaluated after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the evaluation is performed for 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, 18 months or longer, or 24 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor. In some embodiments, one or more of the subsequent doses are higher than the first dose. In some embodiments, one or more of the subsequent doses are lower than the first dose. In some embodiments, one or more subsequent doses are determined based on the creatine kinase activity level evaluated after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, one or more subsequent doses are determined based on the creatine kinase activity level evaluated after administering a previous dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the pre-treatment creatine kinase activity level is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0076] 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 caused by the activity. Relative to an untreated subject performing the same activity, Compound 1 or a salt thereof can reduce the CK level in the subject. The CK level can be measured in the peripheral blood of the subject during or after the activity. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can reduce the CK level in an active subject by 5% to 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can modulate the CK level by about 5% to about 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can reduce the CK level by at least about 5%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can modulate the CK level by up to about 90%. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can reduce the CK level 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 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 caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitors described herein can modulate the CK level by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity.

[0077] Treatment with fast myosin (type II) inhibitors (such as compound 1) of varying durations can reduce the level of creatine kinase activity in a subject. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 1 month to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 2 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 3 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 4 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 5 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 6 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 7 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 8 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level in the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more.In some embodiments, a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 9 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 10 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 11 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more. In some embodiments, a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject for 12 months to maintain the creatine kinase activity level relative to the pre-treatment creatine kinase activity level or to reduce the creatine kinase activity level of the subject by 10% or more, 20% or more, 30% or more, 40% or more, or 50% or more.

[0078] In certain embodiments, an amount of a fast-twitch myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered, the amount maintaining or increasing the lean body mass % of a subject as determined by dual energy x-ray absorptiometry (DXA). In some embodiments, the lean body mass of the subject increases 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 lean body mass of the subject is evaluated after administering a first dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the evaluation is performed 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, 18 months or longer, or 24 months or longer after administering a first dose of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the administration includes 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, one or more subsequent doses are higher than the first dose. In some embodiments, one or more subsequent doses are lower than the first dose. In some embodiments, one or more subsequent doses are determined based on the lean body mass level evaluated after administering a 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 lean body mass level evaluated after administering one or more previous doses of the fast-twitch myosin (type II) inhibitor to the subject. In some embodiments, the pre-treatment lean body mass level is evaluated within 1 month before administering a first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0079] Administering Compound 1 or a salt thereof to a subject can modulate the level of circulating skeletal muscle fast 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 increase after administering an inhibitor, relative to the pre-treatment level of fS-TnI in the subject. Alternatively, the level of fS-TnI in peripheral blood can decrease after administering an inhibitor, relative to the pre-treatment level of fS-TnI in the subject. Administration of the inhibitors described herein can modulate the fS-TnI level by 5% to 90% relative to the pre-treatment level of fS-TnI in the subject. In some cases, the fS-TnI level can be modulated by at least about 5% relative to the pre-treatment level of fS-TnI in the subject. In some cases, the fS-TnI level can be modulated by at most about 90% relative to the pre-treatment level of fS-TnI in the subject. In some cases, the fS-TnI level can be modulated 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 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 fS-TnI level can be modulated by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85% or about 90% relative to the pre-treatment level of fS-TnI in the subject.

[0080] The isoforms of troponin can be measured in a subject before and after administration of Compound 1 or a salt thereof. Inhibiting 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, inhibiting skeletal muscle contraction may not significantly inhibit cTnI or ssTnI. As used herein, with respect to cTnI or ssTnI, the phrase “not significantly” means that the reduction in cTnI or ssTnI is 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% relative to cTnI or ssTnI before administration of the inhibitor.

[0081] In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered that is sufficient to maintain or decrease the TNNI2 concentration in the subject relative to the pre-treatment TNNI2 concentration. 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 administration of a first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the evaluation is performed 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, 18 months or longer, or 24 months or longer after administration of a first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the administration includes administration of a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor. In some embodiments, one or more of the subsequent doses are higher than the first dose. In some embodiments, one or more of the subsequent doses are lower than the first dose. In some embodiments, one or more subsequent doses are determined based on the TNNI2 level evaluated after administration of a first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, one or more subsequent doses are determined based on the TNNI2 level evaluated after administration of one or more previous doses of the fast myosin (type II) inhibitor to the subject. In some embodiments, the pre-treatment TNNI2 level is evaluated within 1 month before administration of a first dose of the fast myosin (type II) inhibitor to the subject.

[0082] Treatment with fast myosin (type II) inhibitors (such as compound 1) for different durations can reduce the TNNI2 level in a subject. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 1 month to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 2 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 3 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 4 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 5 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as compound 1) is administered to the subject for 6 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or reduce the subject's TNNI2 level 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.In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 7 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 8 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 9 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 10 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 11 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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. In some embodiments, a fast myosin (type II) inhibitor (such as Compound 1) is administered to a subject for 12 months to maintain the subject's TNNI2 level relative to the pre-treatment TNNI2 level or to reduce the subject's TNNI2 level 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.

[0083] Elevated serum myoglobin levels can be found in patients with several types of neuromuscular conditions, and thus myoglobin can be a potential indicator for assessing the progression of neuromuscular conditions. In some embodiments, a quantity of a fast myosin (type II) inhibitor, such as Compound 1, is administered to an object, the quantity being sufficient to maintain the myoglobin level or reduce the myoglobin level of the object 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 relative to the pre-treatment myoglobin level. In some embodiments, a quantity of a fast myosin (type II) inhibitor, such as Compound 1, is administered to an object, the quantity being sufficient to maintain the myoglobin level or reduce the myoglobin level of the object 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 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% relative to the pre-treatment myoglobin level. In some embodiments, a quantity of a fast myosin (type II) inhibitor, such as Compound 1, is administered to an object, the quantity being sufficient to maintain the myoglobin level or reduce the myoglobin level of the object by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90% relative to the pre-treatment myoglobin level.

[0084] In some embodiments, the myoglobin level of a subject is evaluated after administering a first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the evaluation is performed for 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, 18 months or longer, or 24 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor. In some embodiments, one or more of the subsequent doses are higher than the first dose. In some embodiments, one or more of the subsequent doses are lower than the first dose. In some embodiments, one or more subsequent doses are determined based on the myoglobin level evaluated after administering the first dose of the fast myosin (type II) inhibitor to the subject. In some embodiments, one or more subsequent doses are determined based on the myoglobin level evaluated after administering one or more previous doses of the fast myosin (type II) inhibitor to the subject. In some embodiments, the pre-treatment myoglobin level is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0085] 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 strenuous activity to reduce or prevent skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, Compound 1 or a salt thereof can reduce the myoglobin level in the subject. The myoglobin level in the peripheral blood of the subject can be measured during or after the activity. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can reduce the myoglobin level in an active subject by 5% to 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can modulate the myoglobin level by about 5% to about 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can reduce the myoglobin level by at least about 5%, thereby reducing or preventing skeletal muscle breakdown caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can modulate the myoglobin level by up to about 90%. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can reduce the myoglobin level 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 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 caused by the activity. Relative to an untreated subject performing the same activity, administration of the inhibitor described herein can modulate the myoglobin level by about 5%, about 15%, about 25%, about 35%, about 45%, about 55%, about 65%, about 75%, about 85%, or about 90%, thereby reducing or preventing skeletal muscle breakdown caused by the activity.

[0086] In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered, the amount being sufficient to maintain or reduce the TNNI2 concentration in a subject relative to the pre-treatment TNNI2 concentration. In some embodiments, the TNNI2 concentration is reduced by 120 ng / mL or more, 100 ng / mL or more, 80 ng / mL or more, 65 ng / mL or more, 50 ng / mL or more, 45 ng / mL or more, 40 ng / mL or more, 35 ng / mL or more, 30 ng / mL or more, 25 ng / mL or more, 20 ng / mL or more, 15 ng / mL or more, 10 ng / mL or more, 5 ng / 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, 5 to 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.

[0087] In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration in a subject to about 1 ng / mL to about 50 ng / mL. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration in a subject 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, about 3 ng / mL to about 5 ng / mL, about 3 ng / mL to about 8 ng / mL, about 3 ng / mL to about 10 ng / mL, about 3 ng / mL to about 15 ng / mL, about 3 ng / mL to about 20 ng / mL, about 3 ng / mL to about 25 ng / mL, about 3 ng / mL to about 30 ng / mL, about 3 ng / mL to about 35 ng / mL, about 3 ng / mL to about 40 ng / mL, about 3 ng / mL to about 50 ng / mL, about 5 ng / mL to about 8 ng / mL, about 5 ng / mL to about 10 ng / mL, about 5 ng / mL to about 15 ng / mL, about 5 ng / mL to about 20 ng / mL, about 5 ng / mL to about 25 ng / mL, about 5 ng / mL to about 30 ng / mL, about 5 ng / mL to about 35 ng / mL, about 5 ng / mL to about 40 ng / mL, about 5 ng / mL to about 50 ng / mL, about 8 ng / mL to about 10 ng / mL, about 8 ng / mL to about 15 ng / mL, about 8 ng / mL to about 20 ng / mL, about 8 ng / mL to about 25 ng / mL, about 8 ng / mL to about 30 ng / mL, about 8 ng / mL to about 35 ng / mL, about 8 ng / mL to about 40 ng / mL, about 8 ng / mL to about 50 ng / mL, about 10 ng / mL to about 15 ng / mL, about 10 ng / mL to about 20 ng / mL, about 10 ng / mL to about 25 ng / mL, about 10 ng / mL to about 30 ng / mL, about 10 ng / mL to about 35 ng / mL, about 10 ng / mL to about 40 ng / mL, about 10 ng / mL to about 50 ng / mL, about 15 ng / mL to about 20 ng / mL, about 15 ng / mL to about 25 ng / mL, about 15 ng / mL to about 30 ng / mL, about 15 ng / mL to about 35 ng / mL, about 15 ng / mL to about 40 ng / mL,From about 15 ng / mL to about 50 ng / mL, from about 20 ng / mL to about 25 ng / mL, from about 20 ng / mL to about 30 ng / mL, from about 20 ng / mL to about 35 ng / mL, from about 20 ng / mL to about 40 ng / mL, from about 20 ng / mL to about 50 ng / mL, from about 25 ng / mL to about 30 ng / mL, from about 25 ng / mL to about 35 ng / mL, from about 25 ng / mL to about 40 ng / mL, from about 25 ng / mL to about 50 ng / mL, from about 30 ng / mL to about 35 ng / mL, from about 30 ng / mL to about 40 ng / mL, from about 30 ng / mL to about 50 ng / mL, from about 35 ng / mL to about 40 ng / mL, from about 35 ng / mL to about 50 ng / mL, or from about 40 ng / mL to about 50 ng / mL. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration in a subject 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 40 ng / mL, or about 50 ng / mL. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration in a subject 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, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the TNNI2 concentration in a subject 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.

[0088] In some embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject, the amount being sufficient to reduce the serum creatine kinase (CK) level by 10 U / L or more, 20 U / L or more, 30 U / L or more, 40 U / L or more, 50 U / L or more, 60 U / L or more, 70 U / L or more, 80 U / L or more, 90 U / L or more, 100 U / L or more, 200 U / L or more, 300 U / L or more, 400 U / L or more, 500 U / L or more, 600 U / L or more, 700 U / L or more, 800 U / L or more, 900 U / L or more, 1000 U / L or more, 1100 U / L or more, 1200 U / L or more, 1300 U / L or more, 1400 U / L or more, 1500 U / L or more. In some embodiments, the CK level is reduced by between about 500 U / L and 1500 U / L, between about 800 U / L and 1400 U / L, between about 1000 U / L and 1300 U / L, or between about 1100 U / L and 1300 U / L.

[0089] In certain embodiments, a quantity of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the serum creatine kinase (CK) level of a subject to from about 400 U / L to about 1,500 U / L. In certain embodiments, a quantity of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the serum creatine kinase (CK) level of a subject to from about 400 U / L to about 500 U / L, from about 400 U / L to about 600 U / L, from about 400 U / L to about 700 U / L, from about 400 U / L to about 800 U / L, from about 400 U / L to about 900 U / L, from about 400 U / L to about 1,000 U / L, from about 400 U / L to about 1,100 U / L, from about 400 U / L to about 1,200 U / L, from about 400 U / L to about 1,300 U / L, from about 400 U / L to about 1,400 U / L, from about 400 U / L to about 1,500 U / L, from about 500 U / L to about 600 U / L, from about 500 U / L to about 700 U / L, from about 500 U / L to about 800 U / L, from about 500 U / L to about 900 U / L, from about 500 U / L to about 1,000 U / L, from about 500 U / L to about 1,100 U / L, from about 500 U / L to about 1,200 U / L, from about 500 U / L to about 1,300 U / L, from about 500 U / L to about 1,400 U / L, from about 500 U / L to about 1,500 U / L, from about 600 U / L to about 700 U / L, from about 600 U / L to about 800 U / L, from about 600 U / L to about 900 U / L, from about 600 U / L to about 1,000 U / L, from about 600 U / L to about 1,100 U / L, from about 600 U / L to about 1,200 U / L, from about 600 U / L to about 1,300 U / L, from about 600 U / L to about 1,400 U / L, from about 600 U / L to about 1,500 U / L, from about 700 U / L to about 800 U / L, from about 700 U / L to about 900 U / L, from about 700 U / L to about 1,000 U / L, from about 700 U / L to about 1,100 U / L, from about 700 U / L to about 1,200 U / L, from about 700 U / L to about 1,300 U / L, from about 700 U / L to about 1,400 U / L, from about 700 U / L to about 1,500 U / L, from about 800 U / L to about 900 U / L, from about 800 U / L to about 1,000 U / L, from about 800 U / L to about 1,100 U / L, from about 800 U / L to about 1,200 U / L, from about 800 U / L to about 1,300 U / L, from about 800 U / L to about 1,400 U / L, from about 800 U / L to about 1,500 U / L, from about 900 U / L to about 1,000 U / L, from about 900 U / L to about 1,100 U / L, from about 900 U / L to about 1,200 U / L, from about 900 U / L to about 1,300 U / L, from about 900 U / L to about 1,400 U / L, from about 900 U / L to about 1,500 U / L, from about 1,000 U / L to about 1,100 U / L, from about 1,000 U / L to about 1,200 U / L, from about 1,000 U / L to about 1,300 U / L, from about 1,000 U / L to about 1,400 U / L, from about 1,000 U / L to about 1,500 U / L, from about 1,100 U / L to about 1,200 U / L, from about 1,100 U / L to about 1,300 U / L, from about 1,100 U / L to about 1,400 U / L, from about 1,100 U / L to about 1,500 U / L, from about 1,200 U / L to about 1,300 U / L, from about 1,200 U / L to about 1,400 U / L, from about 1,200 U / L to about 1,500 U / L, from about 1,300 U / L to about 1,400 U / L, from about 1,300 U / L to about 1,500 U / L, or from about 1,400 U / L to about 1,500 U / L. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the serum creatine kinase (CK) level of a subject 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, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the serum creatine kinase (CK) level of a subject 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, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the serum creatine kinase (CK) level of a subject to at most 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.,

[0090] In some embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject, the amount being sufficient to reduce the myoglobin level 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, 1000 ng / mL or more.

[0091] In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the myoglobin level in a subject to from about 100 U / L to about 400 U / L. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the myoglobin level in a subject to from about 100 U / L to about 150 U / L, from about 100 U / L to about 200 U / L, from about 100 U / L to about 250 U / L, from about 100 U / L to about 300 U / L, from about 100 U / L to about 350 U / L, from about 100 U / L to about 400 U / L, from about 150 U / L to about 200 U / L, from about 150 U / L to about 250 U / L, from about 150 U / L to about 300 U / L, from about 150 U / L to about 350 U / L, from about 150 U / L to about 400 U / L, from about 200 U / L to about 250 U / L, from about 200 U / L to about 300 U / L, from about 200 U / L to about 350 U / L, from about 200 U / L to about 400 U / L, from about 250 U / L to about 300 U / L, from about 250 U / L to about 350 U / L, from about 250 U / L to about 400 U / L, from about 300 U / L to about 350 U / L, from about 300 U / L to about 400 U / L, or from about 350 U / L to about 400 U / L. In certain embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the myoglobin level in a subject 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, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the myoglobin level in a subject 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, an amount of a fast myosin (type II) inhibitor, such as Compound 1 or a salt thereof, is administered in an amount sufficient to maintain or reduce the myoglobin level in a subject to at most 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.

[0092] In some embodiments, an amount of a fast myosin (type II) inhibitor, such as Compound 1, is administered to a subject in an amount sufficient to increase the serum creatinine level 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.

[0093] Administration of Compound 1 or a salt thereof can reduce involuntary muscle contractions. The involuntary muscle contractions can be reduced by 20% to 90% relative 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% relative to the involuntary muscle contractions before treatment. In some cases, the involuntary muscle contractions can be reduced by at most about 90% relative to the involuntary muscle contractions before treatment. In some cases, the involuntary muscle contractions can be 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% to 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, the involuntary muscle contractions can be reduced by about 20%, about 25%, about 30%, about 40%, about 50%, about 70%, about 75%, about 80%, about 85%, or about 90% relative to the involuntary muscle contractions before treatment.

[0094] Compound 1 or a salt thereof can be used to improve the activities of daily living (ADL) or habitual physical activity of a subject because mature, functionally unimpaired muscle can be restored. Examples of ADL or habitual activities include, but are not limited to, stair climbing, time to get out of bed, timed chair rise, habitual walking speed, North Star Ambulatory Assessment (NSAA), incremental / endurance shuttle walk, and 6-minute walk distance test. The level or ability of ADL or habitual physical activity can be measured before and after administration of a skeletal muscle inhibitor. Inhibiting skeletal muscle contraction may not affect ADL or habitual physical activity. In some cases, inhibiting skeletal muscle contraction may not significantly affect ADL or habitual physical activity. As used herein, with respect to ADL or habitual physical activity, the phrase "not significantly" means that the level of ADL or habitual activity is 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% relative to the ADL or habitual activity before administration of the inhibitor.

[0095] In some embodiments, administration of Compound 1 or a salt thereof affects the ADL or habitual physical activity of a subject. In some embodiments, the North Star Ambulatory Assessment is used to evaluate the progression of a subject's neuromuscular disease over time. For example, Figure 1Shows the change over time in the NSAA score of human subjects diagnosed with Becker muscular dystrophy. For patients aged 10 to 32 years, the estimated mean annual decrease was -1.22. In some embodiments, administering Compound 1 or a salt thereof slows the rate of progression of the neuromuscular disease as determined by the NSAA score. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -1.22. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -1. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.5. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.4. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.3. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.2. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than -0.1. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0.1. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0.2. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0.3. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0.4. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 0.5. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 1.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 2.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 3.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 4.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 5.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean annual NSAA score to greater than 10.0. In some embodiments, administering Compound 1 or a salt thereof increases the mean 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.

[0096] In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to about 10. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to 2. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to 1.5. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to 1. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to 0.5. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to 0. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1.2 to -0.5. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1 to 2. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -0.5 to 2. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about 0 to 5. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -1 to 5. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about -0.5 to 4. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about 0 to 3. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than about 2 to 10. In some embodiments, administration of a fast myosin (type II) inhibitor increases the NSAA score of a patient to greater than 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 about 10.

[0097] In some embodiments, after administration of a muscle myosin inhibitor such as Compound 1, the NSAA score of a patient increases. In some embodiments, a fast muscle myosin (type II) inhibitor is administered for about 1 month to about 24 months. In some embodiments, after administration of a muscle myosin inhibitor such as Compound 1, the NSAA score of a patient increases. In some embodiments, a fast muscle myosin (type II) inhibitor is administered for about 2 months to about 12 months. In some embodiments, after administration of a muscle myosin inhibitor such as Compound 1, the NSAA score of a patient increases. In some embodiments, a fast muscle myosin (type II) inhibitor is administered for about 4 months to about 12 months. In some embodiments, after administration of a muscle myosin inhibitor such as Compound 1, the NSAA score of a patient increases. In some embodiments, a fast muscle myosin (type II) inhibitor is administered for about 6 months to about 18 months. In some embodiments, after administration of a muscle myosin inhibitor such as Compound 1, the NSAA score of a patient increases. In some embodiments, a fast muscle myosin (type II) inhibitor is administered for about 2 months to about 6 months. In some embodiments, a fast muscle 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, a fast muscle 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, a fast muscle myosin (type II) inhibitor is administered for at most 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.

[0098] An amount of a fast muscle myosin (type II) inhibitor, such as Compound 1, is administered to a subject, the amount being sufficient to maintain or improve one or more functional metrics of the subject. In some embodiments, the functional metrics are selected from the 100-meter timing test speed relative to the 100-meter timing test speed before treatment; the 4-step stair climb time speed relative to the 4-step stair climb time speed before treatment; the 10-meter walk / run speed relative to the 10-meter walk / run speed before treatment; the maximum elbow flexion strength relative to the maximum elbow flexion strength before treatment; the maximum knee extension strength relative to the maximum knee extension strength before treatment; and the maximum grip strength relative to the maximum grip strength value before treatment. In some embodiments, the functional metrics include an assessment of the patient's ability to perform the Gower's maneuver, walk, rise from a chair, stand on one leg, step up onto a box, step down from a box, sit down, rise from the ground, lift the head, stand on the heels, jump double-footed, jump single-footed, or run.

[0099] In some embodiments, an amount of a fast myosin (type II) inhibitor is administered, the amount being sufficient, relative to pre-treatment values, to maintain one or more functional metrics in a subject. In some embodiments, an amount of a fast myosin (type II) inhibitor is administered, the amount being sufficient, relative to pre-treatment values, to improve one or more functional metrics in a subject. In some embodiments, one or more functional metrics are maintained within 20%, 30%, 40%, or 50% of pre-treatment values. In some embodiments, one or more functional metrics are maintained for 2 months or longer, 4 months or longer, 6 months or longer, 8 months or longer, 12 months or longer, 18 months or longer, 24 months or longer, or 36 months or longer.

[0100] The skeletal muscle contraction or force of a subject can be measured before and after administration of Compound 1 or a salt thereof. Such measurements can be made to generate a dose-response curve for Compound 1 or a salt thereof. The dose of Compound 1 or a salt thereof can be adjusted by about 5% to 50% relative to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dose of a skeletal muscle contraction inhibitor can be adjusted by at least about 5% relative to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dose of a skeletal muscle contraction inhibitor can be adjusted by at most about 50% relative to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dose of a skeletal muscle contraction inhibitor can be adjusted by 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 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% relative to the dose that reduces type II skeletal muscle contraction by 90%. In some cases, the dose of a 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% relative to the dose that reduces type II skeletal muscle contraction by 90%. The skeletal muscle contraction can be measured by a muscle strength test after nerve stimulation using a surface electrode (e.g., plantar flexion after peroneal nerve stimulation of the leg), isolated limb assay, heart rate monitor, or activity monitor or an equivalent thereof before and after administration of the skeletal muscle contraction inhibitor.

[0101] The tidal volume in the lungs of a subject can be measured before and after administration of Compound 1 or a salt thereof. Administration may not inhibit the tidal volume in the lungs. In some cases, administration may not significantly inhibit the tidal volume in the lungs. In certain embodiments regarding the tidal volume in the lungs, the phrase "not significantly" means that the tidal volume in the lungs is 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% relative to the tidal volume in the lungs before administration of an inhibitor. The forced expiratory volume in one second test (FEV1) or forced vital capacity test (FVC) or an equivalent test can be used to measure the tidal volume in the lungs of a subject.

[0102] The smooth muscle contraction of a subject can be measured before and after administration of a skeletal muscle contraction inhibitor. Inhibiting skeletal muscle contraction may not inhibit smooth muscle contraction. In some cases, inhibiting skeletal muscle contraction may not significantly inhibit smooth muscle contraction. As used herein, regarding smooth muscle contraction, the phrase "not significantly" means that the smooth muscle contraction is 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% relative to the smooth muscle contraction before administration of the inhibitor. The smooth muscle contraction of a subject can be evaluated by measuring the blood pressure of the subject.

[0103] The neuromuscular coupling of a subject can be measured before and after administration of Compound 1 or a salt thereof. Inhibiting skeletal muscle contraction with the inhibitors described herein may not impair nerve conduction, neurotransmitter release, or electrical depolarization in the skeletal muscles of the subject. In some cases, inhibiting skeletal muscle contraction may not significantly impair the neuromuscular coupling of the subject. As used herein, regarding neuromuscular coupling, the phrase "not significantly" means that the level of neuromuscular coupling in the subject is 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% relative to the level of neuromuscular coupling in the subject before administration of the inhibitor. The neuromuscular coupling in the subject can be evaluated by measuring the nerve-induced electrical depolarization of the skeletal muscle, which is performed by using surface electrodes or needle electrodes and recording the electrical activity generated by the skeletal muscle after electrical stimulation or voluntary stimulation with electromyography (EMG).

[0104] Pain may be associated with the neuromuscular conditions disclosed herein. Accordingly, methods for alleviating pain associated with neuromuscular conditions are disclosed herein, which include administering a fast 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.

[0105] In some embodiments, a fast myosin (type II) inhibitor is administered to a subject having a dystrophin gene mutation selected from del45-48, del 45-47, del 48, del x-51, del 45-55, and del 48-49.

[0106] In some embodiments, a subject is administered an amount of Compound 1 sufficient to maintain a plasma PK value 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, a subject is administered an amount of Compound 1 sufficient to maintain a plasma PK value between about 500 ng / mL and 4,000 ng / mL, between about 500 ng / mL and 4,000 ng / mL, between about 1,000 ng / mL and 3,000 ng / mL, between about 1,300 ng / mL and 2,800 ng / mL, between about 1,400 ng / mL and 2,600 ng / mL, between about 2,000 ng / mL and 2,600 ng / mL, or between about 2,200 ng / mL and 2,400 ng / mL.

[0107] In some embodiments, a sufficient amount of Compound 1 is administered to a subject to maintain a plasma concentration of from about 10 ng / mL to about 200 ng / mL. In some embodiments, a sufficient amount of Compound 1 is administered to a subject to maintain a plasma concentration of from about 10 ng / mL to about 20 ng / mL, from about 10 ng / mL to about 30 ng / mL, from about 10 ng / mL to about 40 ng / mL, from about 10 ng / mL to about 50 ng / mL, from about 10 ng / mL to about 60 ng / mL, from about 10 ng / mL to about 70 ng / mL, from about 10 ng / mL to about 80 ng / mL, from about 10 ng / mL to about 90 ng / mL, from about 10 ng / mL to about 100 ng / mL, from about 10 ng / mL to about 150 ng / mL, from about 10 ng / mL to about 200 ng / mL, from about 20 ng / mL to about 30 ng / mL, from about 20 ng / mL to about 40 ng / mL, from about 20 ng / mL to about 50 ng / mL, from about 20 ng / mL to about 60 ng / mL, from about 20 ng / mL to about 70 ng / mL, from about 20 ng / mL to about 80 ng / mL, from about 20 ng / mL to about 90 ng / mL, from about 20 ng / mL to about 100 ng / mL, from about 20 ng / mL to about 150 ng / mL, from about 20 ng / mL to about 200 ng / mL, from about 30 ng / mL to about 40 ng / mL, from about 30 ng / mL to about 50 ng / mL, from about 30 ng / mL to about 60 ng / mL, from about 30 ng / mL to about 70 ng / mL, from about 30 ng / mL to about 80 ng / mL, from about 30 ng / mL to about 90 ng / mL, from about 30 ng / mL to about 100 ng / mL, from about 30 ng / mL to about 150 ng / mL, from about 30 ng / mL to about 200 ng / mL, from about 40 ng / mL to about 50 ng / mL, from about 40 ng / mL to about 60 ng / mL, from about 40 ng / mL to about 70 ng / mL, from about 40 ng / mL to about 80 ng / mL, from about 40 ng / mL to about 90 ng / mL, from about 40 ng / mL to about 100 ng / mL, from about 40 ng / mL to about 150 ng / mL, from about 40 ng / mL to about 200 ng / mL, from about 50 ng / mL to about 60 ng / mL, from about 50 ng / mL to about 70 ng / mL, from about 50 ng / mL to about 80 ng / mL, from about 50 ng / mL to about 90 ng / mL, from about 50 ng / mL to about 100 ng / mL, from about 50 ng / mL to about 150 ng / mL, from about 50 ng / mL to about 200 ng / mL, from about 60 ng / mL to about 70 ng / mL, from about 60 ng / mL to about 80 ng / mL, from about 60 ng / mL to about 90 ng / mL, from about 60 ng / mL to about 100 ng / mL, from about 60 ng / mL to about 150 ng / mL,From about 60 ng / mL to about 200 ng / mL, from about 70 ng / mL to about 80 ng / mL, from about 70 ng / mL to about 90 ng / mL, from about 70 ng / mL to about 100 ng / mL, from about 70 ng / mL to about 150 ng / mL, from about 70 ng / mL to about 200 ng / mL, from about 80 ng / mL to about 90 ng / mL, from about 80 ng / mL to about 100 ng / mL, from about 80 ng / mL to about 150 ng / mL, from about 80 ng / mL to about 200 ng / mL, from about 90 ng / mL to about 100 ng / mL, from about 90 ng / mL to about 150 ng / mL, from about 90 ng / mL to about 200 ng / mL, from about 100 ng / mL to about 150 ng / mL, from about 100 ng / mL to about 200 ng / mL, or from about 150 ng / mL to about 200 ng / mL. In some embodiments, a sufficient amount of Compound 1 is administered to a subject 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, a sufficient amount of Compound 1 is administered to a subject 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, a sufficient amount of Compound 1 is administered to a subject 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.

[0108] Fast myosin (type II) inhibitors can be administered for different durations. In some embodiments, the fast 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 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.

[0109] In some embodiments, Compound 1 is administered. In some embodiments, Compound 1 is administered at a daily dose of from about 1 mg to about 10 mg. In some embodiments, Compound 1 is administered at the following daily doses: from about 1 mg to about 2 mg, from about 1 mg to about 2.5 mg, from about 1 mg to about 3 mg, from about 1 mg to about 4 mg, from about 1 mg to about 5 mg, from about 1 mg to about 6 mg, from about 1 mg to about 7 mg, from about 1 mg to about 7.5 mg, from about 1 mg to about 8 mg, from about 1 mg to about 9 mg, from about 1 mg to about 10 mg, from about 2 mg to about 2.5 mg, from about 2 mg to about 3 mg, from about 2 mg to about 4 mg, from about 2 mg to about 5 mg, from about 2 mg to about 6 mg, from about 2 mg to about 7 mg, from about 2 mg to about 7.5 mg, from about 2 mg to about 8 mg, from about 2 mg to about 9 mg, from about 2 mg to about 10 mg, from about 2.5 mg to about 3 mg, from about 2.5 mg to about 4 mg, from about 2.5 mg to about 5 mg, from about 2.5 mg to about 6 mg, from about 2.5 mg to about 7 mg, from about 2.5 mg to about 7.5 mg, from about 2.5 mg to about 8 mg, from about 2.5 mg to about 9 mg, from about 2.5 mg to about 10 mg, from about 3 mg to about 4 mg, from about 3 mg to about 5 mg, from about 3 mg to about 6 mg, from about 3 mg to about 7 mg, from about 3 mg to about 7.5 mg, from about 3 mg to about 8 mg, from about 3 mg to about 9 mg, from about 3 mg to about 10 mg, from about 4 mg to about 5 mg, from about 4 mg to about 6 mg, from about 4 mg to about 7 mg, from about 4 mg to about 7.5 mg, from about 4 mg to about 8 mg, from about 4 mg to about 9 mg, from about 4 mg to about 10 mg, from about 5 mg to about 6 mg, from about 5 mg to about 7 mg, from about 5 mg to about 7.5 mg, from about 5 mg to about 8 mg, from about 5 mg to about 9 mg, from about 5 mg to about 10 mg, from about 6 mg to about 7 mg, from about 6 mg to about 7.5 mg, from about 6 mg to about 8 mg, from about 6 mg to about 9 mg, from about 6 mg to about 10 mg, from about 7 mg to about 7.5 mg, from about 7 mg to about 8 mg, from about 7 mg to about 9 mg, from about 7 mg to about 10 mg, from about 7.5 mg to about 8 mg, from about 7.5 mg to about 9 mg, from about 7.5 mg to about 10 mg, from about 8 mg to about 9 mg, from about 8 mg to about 10 mg, or from 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 administered at the following daily doses: about 2.5 mg to about 5 mg, about 2.5 mg to about 7.5 mg, about 2.5 mg to about 10 mg, about 2.5 mg to about 12.5 mg, about 2.5 mg to about 15 mg, about 2.5 mg to about 17.5 mg, about 2.5 mg to about 20 mg, about 5 mg to about 7.5 mg, about 5 mg to about 10 mg, about 5 mg to about 12.5 mg, about 5 mg to about 15 mg, about 5 mg to about 17.5 mg, about 5 mg to about 20 mg, about 7.5 mg to about 10 mg, about 7.5 mg to about 12.5 mg, about 7.5 mg to about 15 mg, about 7.5 mg to about 17.5 mg, about 7.5 mg to about 20 mg, about 10 mg to about 12.5 mg, about 10 mg to about 15 mg, about 10 mg to about 17.5 mg, about 10 mg to about 20 mg, about 12.5 mg to about 15 mg, about 12.5 mg to about 17.5 mg, about 12.5 mg to about 20 mg, about 15 mg to about 17.5 mg, about 15 mg to 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.

[0110] In some embodiments, the fast myosin (type II) inhibitor is administered at a constant dose throughout the course of treatment. In some embodiments, the fast myosin (type II) inhibitor is administered at a constant dose for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the fast myosin (type II) inhibitor is administered at a constant dose 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 dose is greater than 1 mg / day, greater than 2.5 mg / day, greater than 5 mg / day, greater than 10 mg / day, greater than 15 mg / day, greater than 20 mg / day, greater than 25 mg / day, greater than 30 mg / day, greater than 35 mg / day, or greater than 40 mg / day. In some embodiments, the constant dose 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 dose 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 dose is about 10 mg / day.

[0111] In some embodiments, the fast myosin (type II) inhibitor is administered at different doses throughout the course of treatment. In some embodiments, the fast myosin (type II) inhibitor is administered at different doses for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some embodiments, the fast myosin (type II) inhibitor is administered at different doses 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 myosin (type II) inhibitor is administered at a decreasing dose throughout the course of treatment. In a specific embodiment, the fast myosin (type II) inhibitor is administered at an increasing dose throughout the course of treatment. In some embodiments, the fast myosin (type II) inhibitor is administered at x mg / day, then (x + 5) mg / day, then further (x + 5 times 2) mg / day, then further (x + 5 times 3) mg / day, then further (x + 5 times 4) mg / day, and finally (x + 5 times y) mg / day, where x can be a number between 1 - 100, 5 - 100, 5 - 50, 5 - 20, or 5 - 10, and y can be a number between 1 - 10. In some embodiments, the fast myosin (type II) inhibitor is administered at x mg / day, then (x + 5) mg / day, then further (x + 10) mg / day, where x can be between 1 - 100, 5 - 100, 5 - 50, 5 - 20, or 5 - 10. In some embodiments, the fast myosin (type II) inhibitor is administered at 10 mg / day, then 15 mg / day, then further 20 mg / day.

[0112] In some embodiments, wherein the fast myosin (type II) inhibitor is administered at different doses throughout the course of treatment, the different doses are administered for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In some specific embodiments, the fast myosin (type II) inhibitor is administered at 10 mg / day for the first and second months, then 15 mg / day for the third to sixth months, and then further 20 mg / day for the seventh to twelfth months.

[0113] Fast twitch myosin (type II) inhibitors can be administered at different doses, which are determined in real time based on one or more biomarkers in the subject and / or the plasma concentration of the fast twitch myosin (type II) inhibitor. In some embodiments, the different doses are determined by 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 different doses 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 twitch myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast twitch myosin (type II) inhibitor in the subject 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 10 ng / ml and 80 ng / ml, between 20 ng / ml and 70 ng / ml, between 40 ng / ml and 60 ng / ml, between 40 ng / ml and 100 ng / ml, between 20 ng / ml and 200 ng / ml, between 20 ng / ml and 150 ng / ml, between 20 ng / ml and 140 ng / ml, between 20 ng / ml and 130 ng / ml, between 20 ng / ml and 120 ng / ml, between 20 ng / ml and 110 ng / ml, between 20 ng / ml and 100 ng / ml, between 30 ng / ml and 200 ng / ml, between 30 ng / ml and 150 ng / ml, between 30 ng / ml and 140 ng / ml, between 30 ng / ml and 130 ng / ml, between 30 ng / ml and 120 ng / ml, between 30 ng / ml and 110 ng / ml, between 30 ng / ml and 100 ng / ml, between 40 ng / ml and 200 ng / ml, 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 myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast 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 myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast 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 myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast myosin (type II) inhibitor in the subject 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 myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast myosin (type II) inhibitor in the subject 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, 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.

[0114] In some embodiments, the fast-twitch myosin (type II) inhibitor is enriched in muscle. Thus, in some embodiments, the fast-twitch myosin (type II) inhibitor is administered at different doses to maintain the muscle concentration of the fast-twitch myosin (type II) inhibitor in the subject at greater than 500 ng / g, greater than 600 ng / g, greater than 700 ng / g, greater than 800 ng / g, greater than 900 ng / g, greater than 1000 ng / g, greater than 1100 ng / g, greater than 1200 ng / g, greater than 1300 ng / g, greater than 1400 ng / g, greater than 1500 ng / g, greater than 2000 ng / g, or greater than 3000 ng / g. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at different doses to maintain the muscle concentration of the fast-twitch myosin (type II) inhibitor in the subject at less than 10000 ng / g, less than 9000 ng / g, less than 8000 ng / g, less than 7000 ng / g, less than 6000 ng / g, less than 5000 ng / g, less than 4000 ng / g, or less than 3000 ng / g. In some embodiments, the fast-twitch myosin (type II) inhibitor is administered at different doses to maintain the muscle concentration of the fast-twitch myosin (type II) inhibitor in the subject at 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.

[0115] The compositions and methods described herein can be considered to be useful as pharmaceutical compositions for administration to a subject in need. The pharmaceutical composition can comprise a fast-twitch myosin (type II) inhibitor and one or more pharmaceutically acceptable carriers, diluents, excipients, stabilizers, dispersants, suspending agents, and / or thickening agents. In some embodiments, the fast-twitch myosin (type II) inhibitor is a compound or a salt thereof described in WO2020097265, WO2020097258, WO2020097266, WO2021231572, WO2021231572, WO2021231546, WO2021231630, and WO2021231615. In some embodiments, the fast-twitch myosin (type II) inhibitor is described in WO2020097266. In some embodiments, the muscle myosin inhibitor is a compound of formula (I):

[0116] Also known as Compound 1.

[0117] A pharmaceutical composition comprising a fast myosin (type II) inhibitor can be formulated using one or more physiologically acceptable carriers, which include excipients and auxiliaries. The formulation can be adjusted according to the selected route of administration. The pharmaceutical composition comprising the compound, salt or conjugate can be manufactured, for example, by lyophilizing the compound, salt or conjugate, mixing, dissolving, emulsifying, encapsulating or entrapping the conjugate. The pharmaceutical composition can also contain the compound, salt or conjugate in free base form or in pharmaceutically acceptable salt form.

[0118] A pharmaceutical composition comprising a fast myosin (type II) inhibitor can include formulating any compound, salt or conjugate with one or more inert, pharmaceutically acceptable excipients or carriers to form a solid, semi-solid or liquid composition. Solid compositions can include, for example, powders, tablets, dispersible granules and capsules, and in some aspects, the solid compositions also contain non-toxic auxiliary substances, such as wetting agents or emulsifying agents, pH buffering agents and other pharmaceutically acceptable additives. Alternatively, the compound, salt or conjugate can be lyophilized or in powder form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0119] A pharmaceutical composition comprising a fast myosin (type II) inhibitor can contain at least one active ingredient (e.g., the compound, salt or conjugate and other medicaments). The active ingredient can be entrapped in, for example, microcapsules prepared by coacervation techniques or by interfacial polymerization (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 in a coarse emulsion.

[0120] The composition and formulation can be sterilized. Sterilization can be accomplished by sterile filtration by filtration.

[0121] A pharmaceutical composition comprising a fast myosin (type II) inhibitor can be formulated for administration as an injection. Non-limiting examples of injectable formulations can include sterile suspensions, solutions or emulsions in an oily or aqueous vehicle. Suitable oily vehicles can include, but are not limited to, lipophilic solvents or vehicles such as fatty oils or synthetic fatty acid esters, or liposomes. The aqueous injectable suspension can contain substances that increase the viscosity of the suspension. The suspension can also contain suitable stabilizers. The injection can be formulated for bolus or continuous infusion. Alternatively, the composition can be lyophilized or in powder form for reconstitution with a suitable vehicle (e.g., sterile pyrogen-free water) before use.

[0122] A pharmaceutical composition comprising a fast-twitch myosin (type II) inhibitor can be formulated as a unit dose injectable form (e.g., solution, suspension, emulsion) in association with a pharmaceutically acceptable parenteral vehicle. Such vehicles can be inherently non-toxic and non-therapeutic. The vehicle can be water, saline, Ringer's solution, dextrose solution, and 5% human serum albumin. Non-aqueous vehicles such as fixed oils and ethyl oleate can also be used. Liposomes can be used as carriers. The vehicle can contain minor amounts of additives such as substances that enhance isotonicity and chemical stability (e.g., buffers and preservatives).

[0123] A pharmaceutical composition comprising a fast-twitch myosin (type II) inhibitor can 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 the subject through the mucosal layer in the oral cavity or esophagus. In another embodiment, the composition is formulated to deliver one or more pharmaceutically active agents to the subject through the mucosal layer in the stomach and / or intestine.

[0124] In one embodiment, a composition comprising a fast-twitch myosin (type II) inhibitor can include a modified release dosage form. Suitable modified release dosage vehicles include, but are not limited to, hydrophilic or hydrophobic matrix devices, water-soluble barrier coatings, enteric coatings, osmotic devices, multiparticulate devices, and combinations thereof. The composition can also contain non-controlled release excipients.

[0125] In another embodiment, a composition comprising a fast-twitch myosin (type II) inhibitor can be provided in an enteric-coated dosage form. These enteric-coated dosage forms can also contain non-controlled release excipients. In one embodiment, the composition is in the form of enteric-coated granules as a controlled release capsule for oral administration. The composition can further contain 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 can further contain glycerol monostearate 40 - 50, hydroxypropyl cellulose, pyridazine, magnesium stearate, methacrylic acid copolymer type C, polysorbate 80, sugar spheres, talc, or triethyl citrate.

[0126] In another embodiment, a composition comprising a fast-twitch myosin (type II) inhibitor is an enteric-coated controlled release tablet for oral administration. The composition can also contain 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.

[0127] It is also possible to prepare sustained-release compositions comprising fast myosin (type II) inhibitors. Examples of sustained-release formulations can include semipermeable matrices of solid hydrophobic polymers that can contain a compound, salt, or conjugate, and these matrices can be in the form of shaped articles (e.g., films or microcapsules). Examples of sustained-release matrices can include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactides, copolymers of L-glutamic acid and γ-ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers (such as LUPRONDEPO TM , i.e., injectable microspheres composed of a lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-(–)-3-hydroxybutyric acid.

[0128] Drug compositions comprising fast 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. The formulation can be a lyophilized formulation or an aqueous solution. At the doses and concentrations used, the acceptable carrier, excipient, and / or stabilizer can be non-toxic to the recipient. Acceptable carriers, excipients, and / or stabilizers can include buffering agents such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives; 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 glycols.

[0129] Drug compositions comprising fast myosin (type II) inhibitors can also contain 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.

[0130] Drug compositions comprising fast myosin (type II) inhibitors can be provided in dosage forms having at least one component that promotes rapid release of the active agent and at least one component that promotes controlled release of the active agent. In additional embodiments, the dosage form can be capable of providing discontinuous release of the compound in the form of at least two consecutive pulses spaced 0.1 to 24 hours apart in time. The composition can contain one or more controlled-release and non-controlled-release excipients, such as excipients suitable for rupturable semipermeable membranes and as swellable substances.

[0131] There is provided a pharmaceutical composition comprising a fast myosin (type II) inhibitor in a dosage form for oral administration to a subject, which comprises one or more pharmaceutically acceptable excipients or carriers encapsulated in an intermediate reactive layer containing an anti-gastric juice polymeric material and an outer anti-gastric juice layer, the material being partially neutralized with a base and having cation exchange capacity.

[0132] The pharmaceutical composition comprising a fast myosin (type II) inhibitor provided herein can be in unit dose form or multiple dose form. As used herein, unit dose form refers to a physically discrete unit suitable for administration to a human or non-human animal subject and individually packaged. Each unit dose may contain a predetermined amount of the active ingredient sufficient to produce the desired therapeutic effect, as well as the required pharmaceutical carrier or excipient. Examples of unit dose forms include, but are not limited to, ampoules, syringes, and individually packaged tablets and capsules. In some embodiments, the unit dose form can be administered in its fractions or multiples. Multiple dose form is a plurality of identical unit dose forms packaged in a single container so that they can be administered in separate unit dose forms. Examples of multiple dose forms include, but are not limited to, vials, bottles containing tablets or capsules, or pint bottles or gallon bottles. In another embodiment, the multiple dose form contains different pharmaceutical active agents.

[0133] The pharmaceutical composition comprising a fast myosin (type II) inhibitor can also be formulated into a modified release dosage form, including immediate release, delayed release, extended release, prolonged release, sustained release, pulsed release, controlled release, extended release, accelerated and rapid release, targeted release, programmed release, and gastric retention dosage forms. 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, N.Y., 2002; vol. 126; the above documents are incorporated herein by reference in their entirety).

[0134] Embodiments

[0135] Embodiment 1. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain the NSAA score of the subject or increase the NSAA score of the subject relative to the pre-treatment North Star Ambulatory Assessment (NSAA) score.

[0136] Embodiment 2. The method according to Embodiment 1, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the NSAA score of the subject relative to the pre-treatment NSAA score.

[0137] Embodiment 3. The method according to Embodiment 2, which comprises administering an amount of a fast myosin (type II) inhibitor for 2 months or longer, 4 months or longer, or 6 months or longer, the amount being sufficient to maintain the NSAA score of the subject within 20% of the pre-treatment NSAA score.

[0138] Embodiment 4. The method according to Embodiment 1, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to increase the NSAA score of the subject by 0.3 points or more relative to the pre-treatment NSAA score.

[0139] Embodiment 5. The method according to Embodiment 4, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to increase the NSAA score of the subject by 1 point or more relative to the pre-treatment NSAA score.

[0140] Embodiment 6. The method according to any one of Embodiments 1 to 5, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally one or more of the subsequent doses are higher than the first dose.

[0141] Embodiment 7. The method according to Embodiment 6, wherein the NSAA score of the subject is evaluated 2 months or longer, 4 months or longer, or 6 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0142] Embodiment 8. The method according to Embodiment 6 or 7, wherein the pre-treatment NSAA score is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0143] Embodiment 9. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain or improve one or more of the following functional measures of the subject: maximum elbow flexion strength relative to pre-treatment maximum elbow flexion strength; maximum knee extension strength relative to pre-treatment maximum knee extension strength; 10-meter walking / running speed relative to pre-treatment 10-meter walking / running speed; 100-meter timed test speed relative to pre-treatment 100-meter timed test speed; 4-step climb time speed relative to pre-treatment 4-step climb time speed; and maximum grip strength relative to pre-treatment maximum grip strength.

[0144] Example 10. 100-meter timing test speed, 100-meter timing test speed, 4-step climbing time speed, 4-step climbing time speed. The method as described in Example 9, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain one or more functional metrics of the subject relative to pre-treatment values.

[0145] Example 11. The method as described in Example 10, which comprises administering an amount of a fast myosin (type II) inhibitor for 2 months or longer, 4 months or longer, or 6 months or longer, the amount being sufficient to maintain one or more functional metrics of the subject within 20% of pre-treatment values.

[0146] Example 12. The method as described in Example 9, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to increase one or more functional metrics of the subject by 20% or more relative to pre-treatment values.

[0147] Example 13. The method as described in Example 12, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to increase one or more functional metrics of the subject by 30% or more relative to pre-treatment values.

[0148] Example 14. The method as described in any one of Examples 9 to 13, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0149] Example 15. The method as described in Example 14, wherein one or more functional metrics of the subject are evaluated 2 months or longer, 4 months or longer, or 6 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0150] Example 16. The method as described in Example 14 or 15, wherein the pre-treatment values are evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0151] Example 17. A method of treating a neuromuscular condition, which comprises administering an amount of a fast myosin (type II) inhibitor to a subject, the amount being sufficient to maintain the creatine kinase activity level of the subject or reduce the creatine kinase activity level of the subject by 10% or more relative to the pre-treatment creatine kinase activity level.

[0152] Embodiment 18. The method according to embodiment 17, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the creatine kinase activity level of the subject relative to the pre-treatment creatine kinase activity level.

[0153] Embodiment 19. The method according to embodiment 18, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the creatine kinase activity level of the subject within 10% of the pre-treatment creatine kinase activity level.

[0154] Embodiment 20. The method according to embodiment 17, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to reduce the creatine kinase activity level of the subject by 15% or more, 20% or more, 25% or more, 30% or more, or 35% or more relative to the pre-treatment creatine kinase activity level.

[0155] Embodiment 21. The method according to any one of embodiments 17 to 20, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0156] Embodiment 22. The method according to embodiment 21, wherein the creatine kinase activity level of the subject is evaluated 2 months or more, 4 months or more, or 6 months or more after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0157] Embodiment 23. The method according to embodiment 21 or 22, wherein the pre-treatment creatine kinase activity level is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0158] Embodiment 24. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain the serum creatinine level of the subject or increase the serum creatinine level of the subject by 10% or more relative to the pre-treatment serum creatinine level.

[0159] Embodiment 25. The method according to embodiment 24, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the serum creatinine level of the subject relative to the pre-treatment serum creatinine level.

[0160] Embodiment 26. The method according to embodiment 25, which comprises administering an amount of a fast myosin (type II) inhibitor for 2 months or longer, 4 months or longer, or 6 months or longer, the amount being sufficient to maintain the serum creatinine level of the subject within 10% of the pre-treatment serum creatinine level.

[0161] Embodiment 27. The method according to embodiment 24, which comprises administering an amount of a fast myosin (type II) inhibitor, the amount being sufficient to increase the serum creatinine level of the subject by 10% or more or 20% or more relative to the pre-treatment serum creatinine level.

[0162] Embodiment 28. The method according to any one of embodiments 24 to 27, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0163] Embodiment 29. The method according to embodiment 28, wherein the serum creatinine level of the subject is evaluated 2 months or longer, 4 months or longer, or 6 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0164] Embodiment 30. The method according to embodiment 28 or 29, wherein the pre-treatment serum creatinine level is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0165] Embodiment 31. A method of treating a neuromuscular condition, which comprises administering an amount of a fast myosin (type II) inhibitor to a subject, the amount being sufficient to maintain the DXA % lean body mass of the subject or increase the DXA % lean body mass of the subject by 5% or more relative to the pre-treatment dual energy x-ray absorptiometry (DXA) % lean body mass.

[0166] Embodiment 32. The method according to embodiment 31, which comprises administering an amount of a fast myosin (type II) inhibitor, the amount being sufficient to maintain the DXA % lean body mass of the subject relative to the pre-treatment DXA % lean body mass.

[0167] Embodiment 33. The method according to embodiment 32, which comprises administering an amount of a fast myosin (type II) inhibitor for 2 months or longer, 4 months or longer, or 6 months or longer, the amount being sufficient to maintain the DXA % lean body mass of the subject within 5% of the pre-treatment DXA % lean body mass.

[0168] Embodiment 34. The method according to embodiment 31, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to increase the DXA % lean body mass of the subject by 5% or more, 10% or more, or 15% or more relative to the pre-treatment DXA % lean body mass.

[0169] Embodiment 35. The method according to any one of embodiments 31 to 34, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0170] Embodiment 36. The method according to embodiment 35, wherein the DXA % lean body mass of the subject is evaluated 2 months or more, 4 months or more, or 6 months or more after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0171] Embodiment 37. The method according to embodiment 35 or 36, wherein the pre-treatment DXA % lean body mass is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0172] Embodiment 38. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain the TNNI2 concentration of the subject or reduce the TNNI2 concentration of the subject by 10% or more relative to the pre-treatment skeletal muscle fast troponin I (TNNI2) concentration, and optionally wherein the TNNI2 concentration is inferred from the SOMAscan level.

[0173] Embodiment 39. The method according to embodiment 38, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the TNNI2 concentration of the subject relative to the pre-treatment TNNI2 concentration.

[0174] Embodiment 40. The method according to embodiment 39, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the TNNI2 concentration of the subject within 10% of the pre-treatment creatine kinase activity level.

[0175] Embodiment 41. The method according to embodiment 38, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to reduce the TNNI2 concentration of the subject 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 relative to the pre-treatment TNNI2 concentration.

[0176] Embodiment 42. The method according to any one of embodiments 38 to 41, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0177] Embodiment 43. The method according to embodiment 42, wherein the TNNI2 concentration of the subject is evaluated 2 months or more, 4 months or more, or 6 months or more after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0178] Embodiment 44. The method according to embodiment 42 or 43, wherein the TNNI2 concentration is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0179] Embodiment 45. The method according to any one of the foregoing embodiments, wherein the fast myosin (type II) inhibitor is a compound of formula (I):

[0180] Embodiment 46. The method according to any one of the foregoing embodiments, wherein the administration comprises administering the fast 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.

[0181] Embodiment 47. The method according to embodiment 46, wherein the administration comprises administering the fast myosin (type II) inhibitor once a day.

[0182] Embodiment 48. The method according to any one of embodiments 1 to 47, wherein the administration comprises administering the fast myosin (type II) inhibitor to the subject 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.

[0183] Embodiment 49. The method according to embodiment 48, wherein the administration comprises administering the fast myosin (type II) inhibitor to the subject for 4 months or more, 5 months or more, 6 months or more, or 7 months or more.

[0184] Embodiment 50. The method according to any one of the foregoing embodiments, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0185] Embodiment 51. The method according to embodiment 50, wherein the neuromuscular condition is Becker muscular dystrophy.

[0186] Embodiment 52. The method according to embodiment 50, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0187] Embodiment 53. The method according to any one of the preceding embodiments, wherein the subject is 18 years of age or younger.

[0188] Embodiment 54. The method according to any one of embodiments 1 to 52, wherein the subject is over 18 years of age.

[0189] Embodiment 55. The method according to any one of the preceding embodiments, wherein prior to said administration, the method further comprises selecting a subject for treatment having one or more of the 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 greater, 30 ng / mL or greater or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA before treatment of -0.1 to -10, or -0.5 to -3; (e) a serum creatinine before treatment of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0190] Embodiment 56. A method of inhibiting fast myosin (type II) in a subject in need thereof, comprising selecting a subject having one or more of the 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 greater, 30 ng / mL or greater or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA before treatment of -0.1 to -10, -0.5 to -3, or -1 to -4; (e) a serum creatinine before treatment of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater; and administering a fast myosin (type II) inhibitor to the subject.

[0191] Fast-twitch myosin (type II) - Embodiment 57. The method according to 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.

[0192] Embodiment 58. The method according to embodiment 56 or 57, wherein the fast-twitch myosin (type II) inhibitor is a compound of formula (I):

[0193] Embodiment 59. The method according to any one of the preceding embodiments, wherein the administration 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.

[0194] Embodiment 60. The method according to embodiment 59, wherein the administration comprises administering the fast-twitch myosin (type II) inhibitor once a day.

[0195] Embodiment 61. The method according to any one of embodiments 56 to 60, wherein the administration comprises administering the fast-twitch myosin (type II) inhibitor for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0196] Embodiment 62. The method according to embodiment 61, wherein the administration comprises administering the fast-twitch myosin (type II) inhibitor for 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0197] Embodiment 63. The method according to any one of embodiments 56 to 62, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0198] Embodiment 64. The method according to embodiment 63, wherein the neuromuscular condition is selected from Becker muscular dystrophy.

[0199] Embodiment 65. The method according to embodiment 63, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0200] Embodiment 66. The method according to any one of embodiments 56 to 65, wherein the subject is 18 years old or younger.

[0201] Embodiment 67. The method according to any one of embodiments 56 to 66, wherein the administration does not negatively affect the cardiac function of the subject.

[0202] Embodiment 68. The method according to any one of embodiments 61 to 67, wherein one or more of characteristics (a)-(g) are improved after said administration.

[0203] Embodiment 69. A unit dose for oral administration, comprising a compound of formula (I) in an amount of about 1 mg to about 20 mg and a pharmaceutically acceptable excipient.

[0204] Embodiment 70. The unit dose according to embodiment 69, comprising a compound of formula (I) in an amount of about 1 mg to about 5 mg, and a pharmaceutically acceptable excipient.

[0205] Embodiment 71. The unit dose according to embodiment 70, comprising a compound of formula (I) in an amount of about 1 mg to about 3 mg, and a pharmaceutically acceptable excipient.

[0206] Embodiment 72. The unit dose according to embodiment 70, comprising a compound of formula (I) in an amount of about 2.5 mg, and a pharmaceutically acceptable excipient.

[0207] Embodiment 73. The unit dose according to any one of embodiments 69 to 72, wherein the unit dose is formulated as a pill or a tablet.

[0208] 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): and a pharmaceutically acceptable excipient.

[0209] Embodiment 75. A method of treating a neuromuscular condition, comprising administering to a subject in need thereof a dose of a compound of formula (I) of 3 mg to 40 mg per day:

[0210] Embodiment 76. The method according to embodiment 75, comprising administering to the subject a dose of a compound of formula (I) of 15 mg to 40 mg per day.

[0211] Embodiment 77. The method according to embodiment 76, comprising administering to the subject a dose of a compound of formula (I) of 15 mg, 20 mg, 25 mg or 30 mg per day.

[0212] Embodiment 78. The method according to embodiment 77, comprising administering to the subject a dose of a compound of formula (I) of 20 mg per day.

[0213] Embodiment 79. The method according to embodiment 75, comprising administering to the subject a dose of a compound of formula (I) of 2 mg to 15 mg per day.

[0214] Embodiment 80. The method according to embodiment 79, which comprises administering to a subject a dose of the compound of formula (I) of 5 mg to 12.5 mg per day.

[0215] Embodiment 81. The method according to embodiment 80, which comprises administering to a subject a dose of the compound of formula (I) of 2 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg or 12.5 mg per day.

[0216] Embodiment 82. The method according to any one of embodiments 75 to 81, wherein the subject is over 18 years old.

[0217] Embodiment 83. The method according to any one of embodiments 75 to 81, wherein the subject is 18 years old or younger.

[0218] Embodiment 84. The method according to any one of embodiments 75 to 83, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0219] Embodiment 85. The method according to embodiment 84, wherein the neuromuscular condition is Becker muscular dystrophy.

[0220] Embodiment 86. The method according to embodiment 84, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0221] Embodiment 87. The method according to any one of embodiments 75 to 86, wherein the administration comprises administering the compound of formula (I) for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0222] Embodiment 88. The method according to embodiment 87, wherein the administration comprises administering the compound of formula (I) for 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0223] Embodiment 89. The method according to embodiment 88, wherein the administration comprises administering the compound of formula (I) for 6 months or longer.

[0224] Embodiment 90. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of the compound of formula (I): said amount being sufficient to maintain the AUC of the subject 24 at 1000 ng / mL or higher.

[0225] Embodiment 91. The method according to embodiment 90, which comprises administering a compound of formula (I) for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0226] Embodiment 92. The method according to embodiment 91, wherein the administration comprises administering a fast myosin (type II) inhibitor for 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0227] Embodiment 93. The method according to any one of embodiments 90 to 92, wherein an amount of the compound of formula (I) is administered, the amount being sufficient to maintain the AUC of the subject 24 at 1200 ng / mL or higher.

[0228] Embodiment 94. The method according to embodiment 93, wherein an amount of the compound of formula (I) is administered, the amount being sufficient to maintain the AUC of the subject 24 at 1400 ng / mL or higher.

[0229] Embodiment 95. The method according to any one of embodiments 90 to 94, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0230] Embodiment 96. The method according to embodiment 95, wherein the neuromuscular condition is Becker muscular dystrophy.

[0231] Embodiment 97. The method according to embodiment 95, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0232] Embodiment 98. The method according to any one of embodiments 90 to 97, wherein the subject is older than 18 years old.

[0233] Embodiment 99. The method according to any one of embodiments 90 to 97, wherein the subject is 18 years old or younger.

[0234] Embodiment 100. The method according to any one of embodiments 90 to 99, wherein the administration does not negatively affect the cardiac function of the subject.

[0235] Embodiment 101. The method according to any one of embodiments 90 to 100, wherein the administration 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.

[0236] Embodiment 102. The method according to Embodiment 101, wherein the administration comprises administering the compound of Formula I once a day.

[0237] Embodiment 103. The method according to any one of Embodiments 90 to 102, wherein prior to said administration, the method further comprises selecting a subject having one or more of the characteristics (a)-(g) for treatment: (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 greater, 30 ng / mL or greater or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA before treatment of -0.1 to -10, or -0.5 to -3; (e) a serum creatinine of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower before treatment; (f) DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0238] Embodiment 104. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to maintain the myoglobin level of the subject or reduce the myoglobin level of the subject by 10% or more relative to the pre-treatment myoglobin level.

[0239] Embodiment 105. The method according to Embodiment 104, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the myoglobin level of the subject relative to the pre-treatment myoglobin level.

[0240] Embodiment 106. The method according to Embodiment 104, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to maintain the myoglobin level of the subject within 10% of the pre-treatment myoglobin level.

[0241] Embodiment 107. The method according to Embodiment 104, which comprises administering an amount of a fast myosin (type II) inhibitor sufficient to reduce the myoglobin level of the subject by 5% or more, 10% or more, 15% or more, 20% or more, or 25% or more relative to the pre-treatment myoglobin.

[0242] Embodiment 108. The method according to any one of Embodiments 104 to 107, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0243] Embodiment 109. The method according to any one of Embodiments 104 to 108, wherein the myoglobin level of the subject is evaluated 2 months or more, 4 months or more, or 6 months or more after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0244] Embodiment 110. The method according to any one of Embodiments 104 to 109, wherein the pre-treatment myoglobin level is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0245] Embodiment 111. A method of alleviating pain associated with a neuromuscular condition, comprising administering a fast myosin (type II) inhibitor to a subject in need thereof.

[0246] Embodiment 112. The method according to Embodiment 111, wherein the pain is diffuse.

[0247] Embodiment 113. The method according to Embodiment 112, wherein the pain is in the spine and calves of the subject.

[0248] Embodiment 114. The method according to any one of Embodiments 111 to 113, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

[0249] Embodiment 115. The method according to Embodiment 114, wherein the neuromuscular condition is Becker muscular dystrophy.

[0250] Embodiment 116. The method according to Embodiment 114, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0251] Embodiment 117. The method according to any one of the foregoing embodiments, wherein the fast myosin (type II) inhibitor is administered for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0252] Embodiment 118. The method according to Embodiment 117, wherein the fast myosin (type II) inhibitor is administered at a constant dose for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0253] Embodiment 119. The method according to Embodiment 118, wherein the constant dose is 10 mg / day, 15 mg / day, or 20 mg / day.

[0254] Embodiment 120. The method according to Embodiment 119, wherein the constant dose is 10 mg / day.

[0255] Embodiment 121. The method according to embodiment 117, wherein the fast myosin (type II) inhibitor is administered at different doses for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0256] Embodiment 122. The method according to embodiment 121, wherein the fast myosin (type II) inhibitor is administered at decreasing doses for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0257] Embodiment 123. The method according to embodiment 121, wherein the fast myosin (type II) inhibitor is administered at increasing doses for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months.

[0258] Embodiment 124. The method according to embodiment 123, wherein the fast myosin (type II) inhibitor is administered at 10 mg / day, then 15 mg / day, and further then 20 mg / day.

[0259] Embodiment 125. The method according to embodiment 124, wherein the fast myosin (type II) inhibitor is administered at 10 mg / day for the first and second months, then 15 mg / day for the third to sixth months, and further then 20 mg / day for the seventh to twelfth months.

[0260] Embodiment 126. The method according to embodiment 121, wherein the fast myosin (type II) inhibitor is administered at different doses to maintain the plasma concentration of the fast myosin (type II) inhibitor in the subject between 10 ng / ml and 100 ng / ml, 15 - 70 ng / ml, 20 ng / ml and 70 ng / ml, or 30 ng / ml and 70 ng / ml.

[0261] Embodiment 127. The method according to embodiment 121, wherein the fast myosin (type II) inhibitor is administered at different doses to maintain the muscle concentration of the fast myosin (type II) inhibitor in the subject at 1,000 - 4,000 ng / g.

[0262] Embodiment 128. The method according to any one of the foregoing embodiments, wherein the fast myosin (type II) inhibitor is enriched in muscle.

[0263] Embodiment 129. The method according to any one of the foregoing embodiments, wherein the fast myosin (type II) inhibitor comprises a compound of formula (I):

[0264] Embodiment 130. A unit dose as described in Embodiment 69, which comprises about 1 mg to about 15 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0265] Embodiment 131. A unit dose as described in Embodiment 69, which comprises about 5 mg to about 15 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0266] Embodiment 132. A unit dose as described in Embodiment 69, which comprises 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.

[0267] Embodiment 133. A unit dose as described in Embodiment 69, which comprises about 2.5 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0268] Embodiment 134. A unit dose as described in Embodiment 69, which comprises about 5 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0269] Embodiment 135. A unit dose as described in Embodiment 69, which comprises about 10 mg of a compound of formula (I), and a pharmaceutically acceptable excipient.

[0270] Embodiment 136. A method as described in Embodiment 75, which comprises administering to a subject a dose of about 1 mg to about 15 mg / day of a compound of formula (I).

[0271] Embodiment 137. A method as described in Embodiment 75, which comprises administering to a subject a dose of about 5 mg to about 15 mg / day of a compound of formula (I).

[0272] Embodiment 138. A method as described in Embodiment 75, which comprises administering to a subject a dose of about 2.5 mg / day, about 5 mg / day, about 7.5 mg / day, about 10 mg / day, about 12.5 mg / day, about 15 mg / day, about 17.5 mg / day, or about 20 mg / day of a compound of formula (I).

[0273] Embodiment 139. A method as described in Embodiment 75, which comprises administering to a subject a dose of about 2.5 mg / day of a compound of formula (I).

[0274] Embodiment 140. A method as described in Embodiment 75, which comprises administering to a subject a dose of about 5 mg / day of a compound of formula (I).

[0275] Embodiment 141. The method according to embodiment 75, which comprises administering to a subject a dose of about 10 mg / day of a compound of formula (I).

[0276] Embodiment 142. A method of treating a neuromuscular condition, which comprises administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to reduce the protein concentration of one or more muscle injury biomarkers in the subject by 10% or more relative to the pre-treatment protein concentration of the one or more muscle injury biomarkers.

[0277] Embodiment 143. The method according to embodiment 142, which comprises administering an amount of a fast myosin (type II) inhibitor 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.

[0278] Embodiment 144. The method according to embodiment 142, which comprises administering an amount of a fast myosin (type II) inhibitor 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 relative to the pre-treatment protein concentration of the one or more muscle injury biomarkers.

[0279] Embodiment 145. The method according to any one of embodiments 142 to 144, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0280] Embodiment 146. The method according to embodiment 145, wherein the protein concentration of one or more muscle injury biomarkers in the subject is evaluated 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 administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0281] Embodiment 147. The method according to embodiment 145 or 146, wherein the protein concentration of one or more muscle injury biomarker levels is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0282] Embodiment 148. The method according to any one of Embodiments 142 to 147, wherein 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.

[0283] Embodiment 149. A method of treating a neuromuscular condition, comprising administering to a subject an amount of a fast-twitch myosin (type II) inhibitor sufficient to reduce the protein concentration of one or more pro-inflammatory proteins in the subject by 10% or more relative to the pre-treatment protein concentration of the one or more pro-inflammatory proteins.

[0284] Embodiment 150. The method according to Embodiment 149, comprising administering an amount of a fast-twitch myosin (type II) inhibitor 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.

[0285] Embodiment 151. The method according to Embodiment 149, comprising administering an amount of a fast-twitch myosin (type II) inhibitor 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 relative to the pre-treatment protein concentration of the one or more pro-inflammatory proteins.

[0286] Embodiment 152. The method according to 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, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0287] Embodiment 153. The method according to Embodiment 152, wherein the protein concentration of one or more pro-inflammatory proteins in the subject is evaluated 2 months or longer, 4 months or longer, 6 months or longer, 8 months or longer, 10 months or longer, or 12 months or longer after administering the first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0288] Embodiment 154. The method according to Embodiment 152 or 153, wherein the protein concentration of one or more pro-inflammatory proteins is evaluated within 1 month before administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0289] Embodiment 155. The method according to any one of Embodiments 149 to 154, wherein the one or more pro-inflammatory proteins include CCL22, CCL5, CXCL10, FGG, IFN-γ, IL3, PPBP, PF4, or a combination thereof.

[0290] Embodiment 156. A method of treating a neuromuscular condition, comprising administering to a subject an amount of a fast myosin (type II) inhibitor sufficient to increase the protein concentration of one or more anti-inflammatory proteins in the subject by 10% or more relative to the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

[0291] Embodiment 157. The method according to Embodiment 156, comprising administering an amount of a fast myosin (type II) inhibitor sufficient to increase the protein concentration of one or more anti-inflammatory proteins in the subject within 10% of the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

[0292] Embodiment 158. The method according to Embodiment 156, comprising administering an amount of a fast myosin (type II) inhibitor 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 relative to the pre-treatment protein concentration of the one or more anti-inflammatory proteins.

[0293] Embodiment 159. The method according to any one of Embodiments 156 to 158, wherein the administration comprises administering a first dose of the fast myosin (type II) inhibitor and one or more subsequent doses of the fast myosin (type II) inhibitor, and optionally wherein one or more of the subsequent doses are higher than the first dose.

[0294] Embodiment 160. The method according to Embodiment 159, wherein the protein concentration of one or more anti-inflammatory proteins in the subject is evaluated 2 months or longer, 4 months or longer, 6 months or longer, 8 months or longer, 10 months or longer, or 12 months or longer after administering the first dose of the fast myosin (type II) inhibitor to the subject.

[0295] Embodiment 161. The method according to embodiment 159 or 160, wherein the protein concentration of one or more anti-inflammatory proteins is evaluated within 1 month before administering the first dose of the fast-twitch myosin (type II) inhibitor to the subject.

[0296] Embodiment 162. The method according to any one of embodiments 156 to 161, wherein the one or more anti-inflammatory proteins include IL10, IL13, IL22, IL37, IL4, or a combination thereof.

[0297] Embodiment 163. The method according to any one of embodiments 142 to 162, wherein the fast-twitch myosin (type II) inhibitor is a compound of formula (I):

[0298] Embodiment 164. The method according to any one of embodiments 142 to 163, wherein the administration includes 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.

[0299] Embodiment 165. The method according to embodiment 164, wherein the administration includes administering the fast-twitch myosin (type II) inhibitor once a day.

[0300] Embodiment 166. The method according to any one of embodiments 142 to 165, wherein the administration includes administering the fast-twitch myosin (type II) inhibitor to the subject for 2 months or longer, 3 months or longer, 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0301] Embodiment 167. The method according to embodiment 166, wherein the administration includes administering the fast-twitch myosin (type II) inhibitor to the subject for 4 months or longer, 5 months or longer, 6 months or longer, or 7 months or longer.

[0302] Embodiment 168. The method according to 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.

[0303] Embodiment 169. The method according to embodiment 168, wherein the neuromuscular condition is Becker muscular dystrophy.

[0304] Embodiment 170. The method according to embodiment 168, wherein the neuromuscular condition is Duchenne muscular dystrophy.

[0305] Embodiment 171. The method according to any one of Embodiments 142 to 170, wherein the subject is 18 years of age or younger.

[0306] Embodiment 172. The method according to any one of Embodiments 142 to 170, wherein the subject is over 18 years of age.

[0307] Embodiment 173. The method according to any one of Embodiments 142 to 172, wherein before administration, the method further comprises selecting a subject having one or more of the characteristics (a)-(g) for treatment: (a) creatine kinase activity level of 1000 U / L or higher, 1200 U / L or higher, 1300 U / L or higher; (b) TNNI2 concentration of 20 ng / mL or greater, 30 ng / mL or greater or 40 ng / mL or greater; (c) dystrophin gene mutation; (d) annual change in NSAA before treatment of -0.1 to -10, or -0.5 to -3; (e) serum creatinine before treatment of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower; (f) DXA% lean body mass < 75%, less than 70%, less than 65%, less than 60%; and (g) self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

[0308] Example

[0309] The present invention is now generally described, and the present invention will be more easily understood by reference to the following examples, which are included only for the purpose of illustrating certain aspects and embodiments of the present invention and are not intended to limit the present invention in any way.

[0310] Example 1 - Human Clinical Trial

[0311] Figures 2 - 8 Depicted are the positive 4-month interim results from an ongoing open-label, single-center study that evaluated the safety, tolerability, and effect on muscle injury biomarkers, as well as the pharmacokinetics (PK), of Compound 1 in adults with BMD. Twelve adults with BMD were enrolled in the ARCH study and initially received a 10 mg dose during the first 2 months of the study, after which the dose was escalated to a 15 mg oral dose of Compound 1 every night. Compound 1 was well tolerated in all participants, with no drug discontinuation or dose reduction. The most common adverse events observed to date have been dizziness, drowsiness, and headache. Subsequently, all eligible patients were escalated to a 20 mg daily dose according to the protocol.

[0312] When evaluated by laboratory assays, treatment with Compound 1 resulted in a significant reduction in key biomarkers of muscle injury. Importantly, after 4 months, CK and skeletal muscle fast troponin I decreased by an average of 29% and 73% respectively (Figure 5 ) Although the reduction in CK was maintained and consistent with the observations at 2 months, the skeletal muscle fast troponin I level continued to decline with continued exposure to Compound 1, and four patients reached the levels observed in unaffected adults. Similar to the observations after 2 months, both CK and skeletal muscle fast troponin I were significantly reduced in the context of typical daily activity levels measured with a pedometer.

[0313] Four months after administration of Compound 1, the NSAA increased by an average of 1.17 points compared to the pre-treatment baseline. Significantly, nine out of twelve participants showed functional improvement or no decline in the NSAA relative to their baseline ( Figure 2 ). The NSAA improvement observed only four months after administration of Compound 1 differed from the trajectory observed in the natural history study reported by Bello et al. (2016) (characterizing one of the most comprehensively studied BMD cohorts, with an annual decline of 1.22 NSAA points). These observations were further confirmed by an independent study by vande Velde et al. (2021) (showing a decline of 2.5 NSAA points over 2 years).

[0314] Example 2 - Continued Human Clinical Trial

[0315] Figure 10 to Figure 13 depicts promising 12-month results from an ongoing open-label, single-center study evaluating the safety, tolerability, and effects on muscle injury biomarkers, as well as the pharmacokinetics (PK) of Compound 1 in adults with BMD. The study was conducted in three cohorts Figures 10A - 10C , Figure 11B and Figures 12A - 12F with different dosing regimens as depicted. One cohort was dosed with 20 mg of Compound 1 for 12 months. A second cohort was dosed with 15 mg of Compound 1 for 6 months. A third cohort was dosed with 10 mg of Compound 1 for 2 months. Compound 1 was well tolerated at all doses, without any serious adverse events (AEs). Therefore, no treatment plans were aborted due to AEs.

[0316] Several biomarkers of muscle injury were tested to evaluate the efficacy of Compound 1 under different dosing regimens. Figures 10A - 10C showed that Compound 1 caused a sustained reduction in creatine kinase (CK), skeletal muscle fast troponin I (TNNI2), and myoglobin. In particular, the 10 mg dose showed near-maximal reduction. When analyzing the changes in biomarkers of individual participants from the 12-month group, individuals with the highest baseline values of CK or TNNI2 showed the greatest reduction in biomarkers by Compound 1, indicating that Compound 1 provides protection from activity-induced injury. See Figures 10D - 10G .

[0317] Based on Figures 9A - 9C the NSAA score was also quantified. Compound 1 stabilized the NSAA score by reversing the natural course trend by approximately 1.6 points and led to a trend of score improvement. See Figure 11A . The individual NSAA responses in the 12 - month group were reflected in Figure 11C , which shows that approximately 75% of the participants in the 12 - month group had the same or better NSAA scores. In contrast, the expected natural course decline in untreated patients was approximately - 1.2 to - 1.3 points.

[0318] In parallel, the plasma concentration of Compound 1 was also monitored in different dosing regimens. See Figure 11B . For the higher - dose group (e.g., the 20 - mg group), there were patients with high exposure or low exposure. Unexpectedly, the plasma concentration range reached by patients with high exposure after dosing at 20 mg / day was more than 4 times the target exposure predicted by the pre - clinical model. It is also worth noting that Compound 1 was enriched in muscle. The target muscle concentration of Compound 1 from patients with lower plasma concentrations was 1,000 - 4,000 ng / g from pre - clinical experiments. Using muscle biopsy data from a Phase 1 multiple ascending dose (MAD) study, this corresponds to approximately 15 - 70 ng / ml in BMD. In other words, Compound 1 was enriched in muscle by approximately 60 - fold compared to plasma.

[0319] In addition, the participants in the 20 - mg group with lower exposure (highlighted by the lower circles in Figure 11D ) had the best - stabilized or improved NSAA scores (see the bars with darker shading in Figure 11C ). In contrast, participants with poor outcomes (e.g., Figure 11C negative changes in NSAA scores in Figure 11D ) were under high exposure to Compound 1 (highlighted by the upper circles in Figure 11D ). Thus, lower exposure to Compound 1 (e.g., 10 mg) may have more desirable functional benefits.

[0320] In addition, functional tests were conducted to evaluate the improvement in muscle function of the participants caused by Compound 1. For example, no statistically significant deterioration was observed in the 10 - m speed (see Figure 12A ), 100 - m speed (see Figure 12B ), or 4 - 4 step - climb time speed (see Figure 12C ). Different parameters of muscle strength were also quantified and evaluated in Figures 12D - 12F . In addition, some participants in the 20 - mg group were observed to show muscle weakness (see, for example, Figures 12C - 12F ).

[0321] Combined with Figures 10A - 10C and Figures 11B - 11DThe data shown demonstrated a reduction in relevant biomarkers (indicating improved activity-induced muscle damage) and a better NSAA score at 10 mg / day. The 20 mg / day dose produced efficacy but also some muscle weakening. Thus, Compound 1 with optimal dose control (e.g., 10 mg / day) can lead to a favorable balance of muscle protection and avoidance of some muscle weakness.

[0322] BMD individuals typically report more diffuse pain in the spine and calves, see Jacques MF et al., Plos ONE (2019). Self-reported pain evaluations were collected and quantified in Figure 13 . After 12 months of treatment with Compound 1, a positive trend in self-reported pain scores was observed. Thus, Compound 1 has been shown to reduce pain associated with BMD.

[0323] Example 3 - Characterization of the Short-Term and Long-Term Proteomic Responses of a Fast Myosin Inhibitor in Skeletal Muscle in Becker Muscular Dystrophy (BMD)

[0324] Changes in muscle injury biomarkers such as creatine kinase (CK) or skeletal muscle fast troponin I (TNNI2) were evaluated in clinical trial participants. Biomarker abundance was measured by plasma proteomics (SOMAscan) during short-term (1 - 2 months), medium-term (3 - 4 months), or long-term (6 - 12 months) treatment. Figure 14A The design of the study is shown. Twelve BMD participants were enrolled (7 of whom participated in the Phase I MAD study (Example 2)), and initially treated with 10 mg of Compound 1 per day, and as shown, the dose was escalated from 10 mg to 20 mg per day. Blood was drawn at pre-dose baseline and then analyzed regularly by SOMAscan, and CK activity or TNNI2 concentration was quantified by activity assay or ELISA. SOMAscan is a modified aptamer-based multi-step assay for high-throughput, sensitive, and unbiased biomarker measurements. Proteins in the sample selectively bind to fluorescent aptamers, which are then measured on a chip array to obtain values proportional to the absolute concentration.

[0325] In this study, SOMAscan was correlated with CK and TNNI2 to allow direct conversion to absolute concentration and used to identify large-scale proteomic differences between short-term and long-term treatment with Compound 1. Figure 14B Validation of CK and TNNI2 is shown, where a strong correlation was observed between SOMAscan and the absolute measurements of CK (left) and TNNI2 (right). Figure 14CCK and TNNI2 in blood quantified by SOMAscan are shown. Circulating CK and TNNI2 are significantly elevated compared to the values observed in healthy individuals at baseline and are reduced by short-term treatment. By the long-term treatment time point, TNNI2 is reduced to near healthy levels. The indicated significance is for all values at treatment time points relative to the pre-dose baseline. Figure 14D and Figure 14E The effect of Compound 1 on muscle injury proteins is shown. SOMAscan was used to analyze the effect of treatment on a set of the following proteins, which were identified as being associated with true contraction-induced skeletal muscle injury ( Figure 14D ). By short-term treatment, these proteins (such as CK and TNNI2) are reduced compared to the pre-dose baseline and show stable levels or further reduction during long-term treatment. As a group, the muscle injury proteins do not differ in their response to short-term and long-term treatment ( Figure 14E ), indicating that treatment with 10 mg of Compound 1 for 1 - 2 months maximizes the therapeutic muscle effect.

[0326] Figure 14F The proteomics of long-term Compound 1 treatment is shown. The following proteins exhibit two distinct clusters of change: proteins that are reduced by long-term treatment; or proteins that are differentiated based on long-term treatment compared to short-term treatment. After short-term and long-term treatment, Cluster A is significantly reduced compared to the pre-treatment baseline and is characterized by several skeletal muscle proteins. In contrast, the proteins in Cluster B respond only after a longer period of Compound 1 treatment. These proteins mainly represent immune and inflammatory pathways.

[0327] Figure 14G and Figure 14H Favorable changes in inflammatory biomarkers under long-term treatment are shown. After long-term Compound 1 treatment, favorable changes in the inflammatory profile of the study participants were observed. Several pro-inflammatory cytokines and chemokines were very significantly reduced after 6 - 12 months, as shown by the decrease in gray scale intensity ( Figure 14G upper inset), while several anti-inflammatory interleukins increased after long-term treatment, as shown by the increase in gray scale intensity ( Figure 14G lower inset). The changes in protein concentration observed after 6 - 12 months were consistent among the study participants ( Figure 14H ).

[0328] As shown in this study, SOMAscan values were strongly correlated with the absolute measurements of CK and TNNI2. CK, TNNI2, and a broader set of muscle injury proteins increased at pre-dose baseline, decreased shortly after starting treatment, and remained significantly decreased under long-term treatment. The short-term proteomics of compound 1 treatment was dominated by skeletal muscle proteins reflecting muscle injury. Additionally, long-term treatment also showed consistent changes in proteins related to inflammation, with pro-inflammatory factors decreasing while anti-inflammatory cytokines increased. Although this study varied in time and exposure, it was expected that these proteins would require a longer time span to observe significant changes and could reflect the normalization of muscle after the continuous reduction of muscle injury. The proteomic profile of treatment with compound 1 showed a rapid and continuous reduction of muscle injury markers, accompanied by long-term changes in the inflammatory environment, and the proteomic profile shifted towards that of healthy individuals.

[0329] While the preferred embodiments of the invention have been shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided by way of example only. Many variations, changes, and substitutions will occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. The following claims are intended to define the scope of the invention and are intended to cover methods and structures within the scope of these claims and their equivalents.

Claims

1. A method for treating neuromuscular conditions, which comprises administering to a subject in need thereof a compound of formula (I) in a dose of 1 mg to 40 mg per day:

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

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

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

5. The method according to claim 1, which comprises administering to the subject a compound of formula (I) in a dose of 1 mg to 20 mg per day.

6. The method according to claim 5, which comprises administering to the subject a compound of formula (I) in 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.

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

8. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 2 mg per day.

9. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 2.5 mg per day.

10. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 5 mg per day.

11. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 7.5 mg per day.

12. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 10 mg per day.

13. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 15 mg per day.

14. The method according to claim 6, which comprises administering to the subject a compound of formula (I) in a dose of 20 mg per day.

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

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

17. The method according to claim 15, which comprises administering to the subject a compound of formula (I) in a dose of 15 mg per day.

18. The method according to claim 15, which comprises administering to the subject a compound of formula (I) in a dose of 20 mg per day.

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

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

21. The method according to claim 19, which comprises administering to the subject a compound of formula (I) in a dose of 7.5 mg per day.

22. The method according to claim 19, which comprises administering to the subject a dose of 5 mg / day of the compound of formula (I).

23. The method according to claim 19, which comprises administering to the subject a dose of 2.5 mg / day of the compound of formula (I).

24. The method according to claim 19, which comprises administering to the subject a dose of 2 mg / day of the compound of formula (I).

25. The method according to claim 19, which comprises administering to the subject a dose of 1 mg / day of the compound of formula (I).

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

27. The method according to any one of claims 1-26, wherein relative to the pre-treatment North Star Ambulatory Assessment (NSAA) score, the administration maintains the NSAA score of the subject or increases the NSAA score of the subject.

28. The method according to any one of claims 1-27, wherein the administration maintains or improves one or more of the following functional metrics of the subject: maximum elbow flexion strength relative to pre-treatment maximum elbow flexion strength; maximum knee extension strength relative to pre-treatment maximum knee extension strength; 10-meter walking / running speed relative to pre-treatment 10-meter walking / running speed; 100-meter timed test speed relative to pre-treatment 100-meter timed test speed; 4-step climb time speed relative to pre-treatment 4-step climb time speed; and maximum grip strength relative to pre-treatment maximum grip strength value.

29. The method according to any one of claims 1-28, wherein relative to the pre-treatment creatine kinase activity level, the administration maintains the creatine kinase activity level or reduces the creatine kinase activity level of the subject by 10% or more.

30. The method according to any one of claims 1-28, wherein relative to the pre-treatment creatine kinase activity level, the administration maintains the creatine kinase activity level or increases the creatine kinase activity level of the subject by 10% or more.

31. The method according to any one of claims 1-30, wherein relative to the pre-treatment dual energy x-ray absorptiometry (DXA) % lean body mass, the administration maintains the DXA % lean body mass of the subject or increases the DXA % lean body mass of the subject by 5% or more.

32. The method according to any one of claims 1-31, wherein relative to the pre-treatment skeletal muscle fast troponin I (TNNI2) concentration, the administration maintains the TNNI2 concentration of the subject or reduces the TNNI2 concentration of the subject by 10% or more, and optionally wherein the TNNI2 concentration is inferred from the SOMAscan level.

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

34. The method according to claim 33, wherein 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.

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

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

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

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

39. The method according to any one of claims 1 to 38, wherein the neuromuscular condition is selected from Becker muscular dystrophy, Duchenne muscular dystrophy, limb-girdle muscular dystrophy, and McArdle's disease.

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

41. The method according to claim 39, wherein the neuromuscular condition is Duchenne muscular dystrophy.

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

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

44. The method according to claim 42, wherein the administration comprises administering the compound of formula (I) for 6 months or longer.

45. The method according to any one of claims 1-44, wherein prior to said administration, the method further comprises selecting an object having one or more of the characteristics (a)-(g) for treatment: (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 greater, 30 ng / mL or greater, or 40 ng / mL or greater; (c) a dystrophin gene mutation; (d) an annual change in NSAA before treatment of -0.1 to -10, or -0.5 to -3; (e) a serum creatinine of 1 mg / dL or lower, 0.8 mg / dL or lower, 0.6 mg / dL or lower before treatment; (f) DXA% lean body mass < 75%, < 70%, < 65%, < 60%; and (g) a self-reported pain score related to muscular dystrophy of 1.5 or greater, or 2 or greater.

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