Pharmaceutical composition comprising 4-(4-((4-aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol for muscle protection in microgravity, hypoxia and aging

Abstract

The present invention is related to a composition comprising a novel chemical compound with the chemical Name "4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol" and its use. The 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol and related compounds are useful for treating microgravity induced damage such as skeletal muscle atrophy and dysfunction as well as cardiac dysfunction and are also useful for human diseases and conditions accompanied by significant loss of muscle mass and reduced endurance capacity, such as immobilized patients, cancer, heart failure, sepsis, diabetes mellitus, muscle genetic diseases and neurodegenerative disorders. In addition, 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and related compounds are useful for treating human diseases related to muscle degeneration and ageing such as sarcopenia and heart failure.

Description

[0001] Description

[0002] Title of Invention: Pharmaceutical composition comprising 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol for muscle protection in microgravity, hypoxia and aging.

[0003] Technical Field

[0004] The present invention is related to a composition comprising a novel chemical compound with the chemical Name ''4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol'' and its use. The 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and related compounds are useful for treating microgravity induced damage such as skeletal muscle atrophy and dysfunction as well as cardiac dysfunction and are also useful for human diseases and conditions accompanied by significant loss of muscle mass and reduced endurance capacity, such as immobilized patients, cancer, heart failure, sepsis, diabetes mellitus, muscle genetic diseases and neurodegenerative disorders. In addition, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and related compounds are useful for treating human diseases related to muscle degeneration and ageing such as sarcopenia and heart failure.

[0005] The present invention relates to a composition comprising a novel chemical compound with the Name ''4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol'' that induces controlled proliferation and subsequent maturation of myoblasts / myocytes, resistance to apoptosis, increased muscle mass and function of atrophic muscles and increased endurance capacity. It also inhibits the process of cellular senescence. The present invention relates to products containing the same, as well as to methods of its use.

[0006] Background of the invention

[0007] Microgravity associated with space flight results in a substantial degeneration of the musculoskeletal system. Thus, during a long-duration space trip or upon returning to Earth, a significant loss of muscle mass (atrophy) and a deterioration of muscle strength and endurance capacity pose a considerable medical risk to astronauts. Astronauts experience well- documented consequences on their muscle tissue (wasting and / or atrophy) as a result of their limited range of motion and lack of muscle use in microgravity. The most common skeletal muscle characteristic linked to microgravity is atrophy, which is manifested as a decrease in myofiber size as well as a loss of muscle mass. Actually, the antigravity muscles that astronauts normally employ on Earth, remain in a typical state of unloading and disuse because they are no longer needed in the absence of gravity. Significant physical deficits, including fatigue and slowed speed, are the result of these changes in muscle fibre size, resting and active tension, contractile velocity, and neuromuscular junction function. In fact, the biological and clinical signs that astronauts experience both during and after space travel appear to be similar to the biochemical and clinical signs of physical frailty that older adults encounter. Thus, microgravity exposure seems to accelerate the aging process especially in skeletal muscles [Life 2022, 72(12): 2139],

[0008] Previous studies on animals exposed to long-term space missions have demonstrated a considerable muscle mass reduction of 30-40%, compared to the ground controls and give important insights of the pathophysiological mechanisms which involve extensive changes in kinase signalling activation and gene expression at molecular level. Allen et al. [J Appl Physiol 2009, 106: 582-595] reported that in the gastrocnemius of mice that were kept flowing on the mid-deck of the space shuttle Endeavour (STS-108 / UF-1) for 11 days a significant decrease in the activation of the Phosphatidylinositol 3-Kinase (PI3K) / Akt / mTOR pathway was found. Similarly, Sandona et al. performed a long-term (91 -days) experiment on mice (sponsored by the Italian Space Agency onboard the International Space Station) demonstrating that this exposure to microgravity resulted in a decrease in Insulin Growth Factor- 1 (IGF-1) expression, a critical regulator of the PI3K / Akt pathway, and a reduction in muscle mass [Life 2022, 72(12): 2139], In brief, the P 13K / Akt pathway controls protein synthesis via mammalian target of rapamycin (mTOR) and glycogen synthase kinase 3[3 (GSK3J3), and protein degradation via regulation of the transcription factors of the forkhead family of transcription factor (FOXO). Baek et al. observed that exposure of C2C12 myoblasts to simulated microgravity decreases myogenesis (proliferation and differentiation of myoblasts) by decreasing Akt activation while at the same time results in muscle degradation by apoptosis and autophagy as a result of FOXO1 upregulation. In line with the above, muscle atrophy in rats undergoing a 17-day space flight (NASA STS-90 NeuroLab) was associated with decreased IGF-II and upregulation of myostatin. Myostatin, a member of the TGF-J3 (Transforming Growth Factor-[3) superfamily, has a significant role in decreasing muscle mass and interfering with muscle repair via activation of signalling pathways such as p38 MAPK (Mitogen Activated Protein Kinase) and SMAD2 / 3 (Mothers against decapentaplegic 2 / 3). Increased expression of pro-inflammatory cytokines such as TNF-a (Tumor Necrosis Factor-a) and interleukin-6 (IL-6) and oxidative stress are also shown to have a role in microgravity induced muscle degeneration. Overall, results from existing studies suggest that unloading caused by microgravity directly affects the myogenic differentiation program by downregulating pathways associated with skeletal muscle differentiation and protein synthesis and upregulating pathways associated with degradation and apoptosis.

[0009] Microgravity can also alter cardiac muscle function by unloading. Unloading results in reduction in circulatory blood volume and physiologic cardiac atrophy. In a small study of four astronauts exposed to the complete elimination of hydrostatic gradients during a spaceflight of 10 days, a decrease in left ventricular (LV) mass by 12 + / - 6.9% (P = 0.07) was observed compared to control subjects as assessed by Cardiac Magnetic Resonance. Studies in animals after short-term spaceflight have shown evidence of decreased cardiac myocyte size, which is consistent with cardiac atrophy [Progress in Cardiovascular Diseases 2023, 81: 33-41], Long duration spaceflight makes heart contraction less efficient, stroke volume (the amount of blood that is being pumped out of the heart) decreases by 17.4% and cardiac output by 12.2%. These changes result in increased pressure of the heart during the relaxation phase known as diastolic dysfunction [Aviation, Space, and Environmental Medicine. 2002, 73-6: 532-536], In addition, during spaceflight, the configuration of the heart changes from being more elliptical on Earth to more spherical in space. The mean spherical index changed by 9.4% — from 2.01 on Earth to 1.82 in microgravity — a finding that verifies the anatomical change of the heart in space [J. Am. Coll. Cardiol. 2014, 63: A 1096], Overall, these changes are similar to the changes observed in patients after acute myocardial infarction and in patients with dilated cardiomyopathy and heart failure. It is of interest that exposure to simulated microgravity in rodents resulted in increased myocardial injury after ischemia / reperfusion indicating that microgravity may exacerbate the damage after acute myocardial infarction [Can J Physiol Pharmacol 2017,95(l):59-71],

[0010] Prolonged bed confinement and immobilization in patients (e.g. orthopaedic patients) and especially in intensive care units (ICU) result in severe mechanical unloading of the musculoskeletal system and heart with detrimental changes similar to prolonged microgravity. These changes include significant loss of muscle mass, size and function which ultimately lead to muscle atrophy and reduced functional capacity and endurance. An interesting study showed by using highly sensitive proteomic analysis that proteomic signatures of mechanical unloading are related to insulin signalling and are similar in spaceflight and bed rest [PNAS Nexus, 2022, 1: 1-14], Many chronic disease patients experience a concurrent loss of lean muscle mass due to a vicious cycle between immobilization and inflammation, metabolism dysregulation and hypoxia. Common illnesses accompanied by muscle atrophy include diabetes, cancers, heart failure, sepsis, muscle genetic diseases and neurodegenerative disorders [Front. Physiol. 2018, 9:235], Using magnetic resonance imaging (MRI), up to 17% and 40% loss in muscle volume and function were seen following 84-day bed rest. Another 90-day study of bed rest reported up to 26% reduction in muscle cross sectional area (mCSA) in healthy males [Bone 2005, 36 : 1019- 1029] . In addition, two week limb immobilization (casting) studies reported reduction in quadriceps muscle volume, mCSA and strength. Moreover, it has been observed that both contractile rate of force development and maximal isometric muscle strength are reduced significantly after 2 weeks of unilateral leg casting. A recent study showed a significant decrease in real-world walking speed after the 60-day bed-rest in 24 healthy subjects. Interestingly, decreased real-world walking speed after bed-rest was observed for all three groups, regardless of the training intervention [npj Microgravity 2024, 10:6],

[0011] In patients admitted to intensive care unit (ICU), a substantial decrease in muscle thickness of 0.84% to 0.98% per day was reported [Scientific Reports 2022,12:16629], The incidence of ICU acquired muscle weakness is up to 80% in critically ill patients with profound impact on prognosis. In a large study including 1127 patients who underwent cardiac surgery with a mean follow-up period of 6.4 years, it was found that the longer the distance covered in the 6min Walk Test before leaving the hospital, the lower the risk of mortality [Scientific Reports 2024; 14:2493], Sepsis-associated muscle wasting (SAMW) is also characterized by decreased muscle mass, reduced muscle fiber size, and decreased muscle strength, resulting in persistent physical disability. The mechanisms are very similar to those occurring in response to microgravity. Inflammatory cytokines such as IU-6, TNF-a and IL- ip, that increase at the onset of sepsis, cause acute muscle wasting [Int J Mol Sci. 2023, 24(5): 5040], Inflammatory cytokines activate a number of apoptotic and degradative pathways, such as p38 MAPK and nuclear factor kappa-light-chain-enhancer of the activated B cells (NF-KB) which result in protein degradation exceeding protein synthesis, leading to muscle wasting in sepsis. Moreover, evidence suggests that caspase-3, autophagy-lysosomal system and ubiquitin proteasome pathway are all involved in protein degradation and muscle degeneration. Initial acute muscle wasting during sepsis leads to bed confinement, immobilization and disuse atrophy creating a vicious cycle.

[0012] Focal adhesion kinase (FAK) is a mechanosensitive non-receptor protein kinase that regulates PI3K activity and subsequently activates Akt-mTOR pathway. During immobilization and disuse, both the impaired IGF-1 signalling and insulin resistance as well as the reduced FAK activation result in reduced activity of Akt-mTOR pathway and play essential roles in reduced protein synthesis and muscle atrophy on earth. In accordance, it has been reported that genetically modified mice that do not express Akt exhibit significant skeletal muscle atrophy as well as growth deficiency. FOXO transcription factors are upregulated in muscle atrophy under disuse conditions and are found to regulate the ubiquitin proteasome pathway and protein catabolism contributing to muscle atrophy. Important evidence also suggests that the activation of calpain and caspase-3 proteases is required for inactivity-induced limb muscle atrophy [J Appl Physiol 2013, 114: 1482-1489],

[0013] Losses in skeletal muscle mass are clearly involved in disuse-induced muscle weakness as muscle cross-sectional area (mCSA) is linearly related to force generating capacity [J Appl Physiol 2022, 132: 835-861], Interestingly, during disuse the loss of strength disproportionately exceeds the loss in muscle mass, implying that losses in strength and power following disuse are also related to changes in muscle fiber types. Disuse-induced mitochondrial deficits may also be contributing to impaired recovery following disuse independent of changes in muscle mass. Regarding neuromuscular function, it has been shown to be decreased following immobilization, which may explain the rapid and elevated losses in strength compared with muscle mass [J Appl Physiol 2022, 132: 835-861],

[0014] Another important element in the process of muscle atrophy is the function of satellite muscle cells which are resident myogenic stem cells with the ability to form new muscle tissue through a stepwise process of proliferation, differentiation, fusion, and maturation after injury. In this regard, 14 days bed rest in healthy adults was associated with reduced satellite cell content [J Appl Physiol 2016, 120: 965-975], Thus, muscle atrophy under disuse conditions includes increased degeneration / apoptosis and decreased repair / regeneration capacity at the same time.

[0015] Longer life expectancy is linked to a higher risk of chronic degenerative diseases, which are commonly seen in older populations. Healthcare systems must bear the enormous financial burden of this evolution. Ageing "per se" has long been thought to be a necessary condition for the emergence of many age-related diseases. Sarcopenia, defined as the age-related decline of muscle mass and function, is undoubtedly one of the most critical adverse effects of ageing and a prevalent problem in this population. Sarcopenia is considered the biological substrate of frailty, a condition of increased vulnerability to stressors. Indeed, ageing is typically characterized by muscle wasting that progressively causes disability, loss of muscle function and of self-sufficiency in older subjects [Life 2022, 12: 2139], Muscle mass reaches its peak between 30 and 40 years of age and starts declining after that, up to a reduction of 25-30% in the cross-sectional area of the skeletal muscle and 40% in muscle strength. Sarcopenia has now received a specific International Classification of Diseases, Tenth Revision (ICD-10) making it a formally recognized disease. Sarcopenia affects approximately 50 million individuals, and this number is projected to surpass 200 million within the next four decades [Age Ageing. 2019,48(4): 601] . The aetiology of sarcopenia is multifactorial, involving many biological mechanisms similar to microgravity induced muscle atrophy. These mechanisms include increased muscle degeneration due to chronic inflammation as well as reduced satellite cell number / function. IGF- 1 / PI3K / Akt pathway reduction have been suggested as a key player in sarcopenia, reduced endurance capacity and frailty. Lower IGF-1 serum levels are linked to reduced handgrip strength and physical performance, a higher risk of impairment, and they are also independently linked to lower skeletal muscle mass in older adults. Moreover, as suggested by several studies, the fate of muscles in older subjects depends mostly on the severity and chronicity of inflammation. An increased expression of IL-6 and TNF-a in skeletal muscle, has been found to result in loss of muscle mass and strength. A report by Kamper and colleagues [Journal of Cachexia, Sarcopenia and Muscle 2021, 12:1641-1652], using data from the Copenhagen Sarcopenia Study, observed that, during ageing, the systemic levels of TNF-a and the C-reactive protein (CRP) increase especially in more physically frail older subjects. Of note, TNF-a plays a crucial role in the pathogenesis of sarcopenia and frailty, since it directly upregulates the NF-KB pathway and the ubiquitin-proteasome system via p38 MAPK signalling activation. This pathway leads to the loss of skeletal muscle proteins and myofibrils degradation.

[0016] Muscle stem cells (known as satellite) reside within muscle tissue in a quiescent state until they are primed to regenerate damaged muscle through cycles of divisions, differentiation, fusion, and maturation. With ageing, satellite cell population and proliferative capacity are known to decline [J Appl Physiol 2016, 120: 965-975], which impairs the ability of the muscle to recover from an injury. The cell -intrinsic dysfunction of the aged satellite cells arises from alterations in signal transduction pathways such as activation of the stressed associated p38 MAPK and JAK / STAT signalling axes. These pathways can negatively regulate self-renewal of satellite cells by restricting cell cycle progression and cell division. Furthermore, the regenerative capacity of aged satellite muscle cells can be recapitulated by inhibition of p38 MAPK and JAK / STAT signalling axes. In particular, rejuvenation of aged satellite muscle cells is achieved after transient treatment with a chemical inhibitor of p38 MAPK [Nat Med 2015,21(8): 854-862],

[0017] Dysfunction of the aged satellite cells indicates the importance of understanding the mechanisms involved in the pathogenesis of ageing-related diseases. Age-related muscle dysfunction fuels several co-morbidities such as diabetes and heart failure and creates a vicious cycle leading to disease progression and death. In particular, age-related changes in skeletal muscle include reduced glucose utilization and increased resistance to insulin and thus contribute to the development of type 2 diabetes. In addition, the onset of diabetes accelerates skeletal muscle loss leading to a vicious cycle [Proceedings of the Nutrition Society 2020, 79: 158-169], Studies document that magnetic resonance scans of 103 cases of diabetic patietns showed edema in 76.8% of cases, while the initial presentation of thigh pain or swelling has been documented in 83.7% and 80% of cases. The pathophysiology of diabetic myopathy involves a microvascular pathological process resulting in inflammation and hypoxia. Diabetes has been shown to alter the function of satellite cells involved in muscle growth and regeneration. Satellite cell function has been proposed to be considerably affected by hyperglycaemic and lipotoxic conditions associated with diabetic state. In the obese and diabetic Zucker rat model, reduced satellite cell proliferative capacity has been documented [frit J Mol Sci 2023, 25( 1):469] .

[0018] There is significant evidence that muscle microcirculation plays an important role during muscle atrophy and regeneration. Capillary rarefaction does not only occur during ageing, but also during conditions as chronic heart failure and diabetes, where endothelial apoptosis has been reported to precede muscle atrophy. It has been reported that capillary rarefaction precedes sarcopenia in older people and may contribute to the age-related muscle atrophy and decline in exercise capacity. Microcirculatory dysfunction is also recognized as a central factor in pathogenesis of critical illness and ICU acquired weakness. Inflammatory activation of the endothelium in critical illness results in secretion of numerous mediators that may subsequently affect muscle function; indeed, inflammatory activation of skeletal muscle via endothelial -myocyte crosstalk is shown to contribute to pathogenesis of the ICU- acquired weakness [Front. Physiol. 2023, 14:1170429], Interestingly, in vitro endothelial cells stimulate satellite cells growth through secretion of growth factors. It has been also reported that in muscle tissue, active satellite cells are located closer to capillaries than quiescent satellite cells [Journal of Muscle Research and Cell Motility 2019, 40:127-140],

[0019] Heart failure (HF) is characterized by abnormal cardiac structure and function, leading to systolic and diastolic dysfunction. Sarcopenia and HF are closely related conditions [ESC Heart Fail. 2018, 5(6): 1074-1082], Sarcopenia is a critical comorbidity in heart failure, according to HF Guidelines published by the European Society of Cardiology. Patients suffering from chronic heart failure (CHF) are increasingly likely to be sarcopenic. A mixed cohort study on symptomatic heart failure patients revealed a 20% prevalence of sarcopenia among the patients, which was much greater than the incidence observed in older persons in good health. Additionally, a multicenter clinical investigation found a clear correlation between reduced exercise ability in HF patients and lower skeletal muscle mass. Furthermore, it has been demonstrated that sarcopenia negatively impacts the prognosis of HF patients and has been found to be an independent risk factor for death in CHF patients (J Cachexia Sarcopenia Muscle. 2020, 11(5): 1242-1249). In accordance, in end stage HF patients with a newly implanted ventricular assist device, the rehospitalization rate was significantly higher in patients with lower leg muscle strength compared to those with higher leg muscle strength. Current research on the pathophysiological mechanisms underlying HF with sarcopenia primarily revolves around inflammation, hormonal changes and oxidative stress.

[0020] From the above, it is obvious that several characteristics of ageing and age-related diseases have similarities with those problems confronted during spaceflight and thus synergies can be drawn between microgravity, ageing and age-related disorders. The strategies used in space for maintenance of astronaut health could also be used for the benefit of life on Earth.

[0021] Currently available therapies to counteract the loss of skeletal muscle from microgravity and ageing and restore muscle strength include resistance exercises, antioxidants and protein supplements. In this regard, physical activity and / or nutritional interventions are considered the currently available strategies to counteract muscle atrophy and bone mineral density in both older adults and astronauts. Exercise has been demonstrated to have a number of benefits for muscles, including a reduction in inflammation, oxidative damage and mitochondrial malfunction, regulation of the balance between muscle protein synthesis and breakdown, and stabilisation of the autophagy processes. Interestingly, it has been described that a multicomponent intervention, based on physical activity and nutritional counselling, is associated with a reduction in the incidence of physical frailty and sarcopenia in older subjects. In this regard, exercise continues to be the major defence against the decline in astronauts' and older adults' physical performance. However, despite 2 h of daily exercise training, muscle wasting is still present after 6-month missions to the international space station. Optimizing countermeasures for astronauts and space travellers is particularly important as plans for prolonged missions to the Moon and to Mars are under way.

[0022] Early rehabilitation has been also introduced in the ICU to reduce the occurrence and severity of muscle weakness in these patients. However, even with early rehabilitation, there is considerable muscle atrophy in ICU patients. In addition, in most cases, remobilization starts too late and patients permanently lose their independence and autonomy; this is associated with higher morbidity and mortality. Supplemental nutrition has been adopted as an additional intervention to prevent low vitamin D status, which appears to be linked to muscle loss and poor performance. It has been tested in both space flight studies and the geriatric population but it is still debatable whether vitamin D supplementation helps prevent muscle atrophy in elderly sarcopenic patients or astronauts. In the clinical setting of SAMW, electrical muscular stimulation, physiotherapy, early mobilization, and nutritional support are used for patients with sepsis to prevent or treat muscle wasting. Despite these measures, SAMW occurs in 40% of critically ill, sepsis patients and is associated with prolonged ventilator use, extended hospital stay, increased mortality, and long-term functional disorders. Although improvement and prevention of SAMW are important issues, there are no approved pharmacological therapeutic compounds for this condition.

[0023] There is currently no approved drug to combat muscle wasting on terrestrial diseases or in microgravity. Only a few pharmacological approaches are tested in preclinical studies fortheir ability to counteract disuse-associated muscle loss, while other pharmacological approaches that have reached initial clinical trials, have failed [Br J Clin Pharmacol. 2023, 1-13], The Sarcoendoplasmic Reticulum Calcium ATPase (SERCA) pump is a key regulator of cellular calcium homeostasis and is closely associated with muscle health and function. Pathological conditions linked to ageing, muscle unloading and disuse, neurodegeneration, and muscular dystrophy have been extensively documented to significantly lower SERCA function. This can potentially lead to an impairment in intracellular calcium homeostasis, which in turn can exacerbate muscle atrophy and weakness [Skelet Muscle 2021, 11(1): 25], Thus, targeting SERCA activity has gained interest as a therapeutic approach to treat muscle diseases. Treatment with CDN1163, an allosteric SERCA activator, prevented age-related muscle atrophy and weakness in mice [Int J Mol Sci. 2021, 22(1): 37],

[0024] Treatment with steroidal androgens has shown some improvement after muscle disuse in small clinical trials but only when combined with exercise. Clinical practice guidelines indicate that the effect of testosterone in men with low serum levels of testosterone is important on muscle mass but it is considered modest or minimal on muscle strength. Most investigations conclude that testosterone benefits on physical performance is unconvincing. In addition, its clinical use is considerably limited by severe side effects including behavioural abnormalities (which could be crucial in the spaceflight context) and amplified risk for developing prostate hypertrophy, cancer, sleep apnoea or thrombosis complications.

[0025] Selective androgen receptor modulators (SARMs) are small molecule drugs selectively targeting androgen receptors to elicit anabolic effects on muscle tissue. In small clinical trials, the SARM GTx-024 showed increased total lean body mass and physical ability. In a larger trial in 170 elderly women with sarcopenia, a SARM MK-0773 for 6 months resulted in a significant increase in muscle mass, even though no differences were found for muscle strength or physical performance. To date, none of the candidate SARMs has been approved for therapeutic use by the Food and Drug Administration and certainly not moved towards the market probably because their use in athletes is associated with serious or life-threatening health problems. Further to that, their effectiveness in both men and women is an issue under question and they have been discredited due to their potential hepatotoxicity [Br J Clin Pharmacol. 2023, 1-13],

[0026] The myokine myostatin is a signalling molecule negatively regulating muscle mass via signalling through activin type II receptors A and B. Therapeutic myostatin inhibition has been purported for muscular dystrophy, cachexia, sarcopenia, and disuse atrophy associated with different diseases. Subsequent development of a range of myostatin inhibitors and promising pre-clinical results encouraged human trials. These drugs have been tested in multiple phase 2 clinical trials for a wide range of indications, focused on muscle loss and the associated functional limitations often seen in older adults and people with chronic diseases. In older weak individuals who had fallen during the previous year, LY2495655 was shown to cause increases in lean body mass, accompanied by a persistent decrease in fat mass, as well as statistical improvements in stair climbing and gait speed compared to placebo, although clinical relevance was unclear. In patients following total hip arthroplasty, LY2495655 failed to achieve statistical significance in endpoints such as the 6-minute walk test. In addition, the trial did not show any improvements compared to the standard of care that included post-operative physical therapy. Bimagrumab, a monoclonal antibody that inhibits the action of Myostatin and Activin A, was tested in 2 trials for sarcopenia in older adults and failed to show consistent improvements in muscle strength and physical function. A similar result of an increase in lean body mass with no improvement in physical function was seen following 6 months of treatment in patients undergoing hip fracture repair surgery. Overall, functional improvements in the older population treated with Myostatin inhibitors have been inconsistent [The Journals of Gerontology 2023, 78(1): 32-37], Phase II clinical trials have also been performed with myostatin inhibitors in patients with muscular dystrophy, neuromuscular disorders and cachexia due to cancer but no improvement in muscle strength and physical function has been demonstrated.

[0027] Apelin (AP) is a small peptide with different isoforms that binds to APJ receptor and activates AMPK (AMP-activated protein kinase)-dependent pathways leading to mitochondriogenesis in skeletal muscles. In aged mice and sarcopenic humans, plasma apelin is decreased while recombinant apelin systemic treatment leads to an increase of muscle mass and function in aged mice by activating metabolism through AMPK and Akt in skeletal muscle. Moreover, APJ is present on satellite cells and its activation in aged mice results in an increase of proliferation of these cells alongside a better regenerative process. Clinical trials with apelin analogs are underway [Metabolism 2023, 149:155597],

[0028] A diverse range of hereditary illnesses, known as muscular dystrophies, are characterised by the progressive deterioration of cardiac and skeletal muscle. Gene mutations encoding various cellular components cause muscle dysfunction in muscular dystrophies, leading to progressive muscle weakness and wasting [J Clin Invest. 2020, 130( 11): 5652-5664] . The age at which symptoms first appear, the degree to which symptoms worsen, and the location of the affected muscles vary amongst dystrophies. Specifically, cardiac involvement is seen in a number of dystrophies. Duchenne muscular dystrophy (DMD) is the most common and one of the most severe forms of muscular dystrophy, occurring in about 1 in 5000 male newborns. Patients with DMD often exhibit progressive weakness in the diaphragm, trunk muscles, and limb muscles, which can result in kyphoscoliosis, wasting, and serious respiratory issues. The majority of patients pass away in their third decade of life mainly due to respiratory complications. Regardless of the underlying pathophysiological mechanisms, dystrophic muscle displays cycles of deterioration and regeneration along with inflammatory cells infiltration and a steady build-up of fibrotic and adipose tissues [J Clin Invest. 2020, 130(11): 5652-5664], Although glucocorticoid treatment is associated with reduced disease progression and multidisciplinary care may further improve patient survival, there is currently no definitive cure for muscular dystrophies. Numerous intriguing therapeutic approaches, such as gene delivery and cell-based therapy, are currently being actively researched. Cell therapy, which uses genetically normal or genetically corrected cells can introduce normal copies of a gene (or other therapeutic genes in patients) into myofibers through cell fusion. Therefore, creating strategies for gene correction and complementation is essential to the effective use of cell therapy. Transplantation of muscle stem or progenitor cells also has the potential to support or improve muscle repair and could cover future regenerative needs. Pharmacological enhancement of the muscular environment in recipient dystrophic muscles as well as in cultured donor cells for infusion could make it easier for cells to engraft. An alternative strategy that might make it possible to replace damaged tissue is artificial, bioengineered, skeletal muscle made by satellite and muscle resident cells [Cells 2019, 8(9): 1066],

[0029] Thus, novel pharmacological interventions that counteract and / or reverse muscle atrophy, reduced strength and endurance both in space and earth related diseases and ageing do not exist. Novel pharmacological interventions with the capacity to inhibit muscle degeneration, and at the same time induce proliferation and maturation of myoblast cells to form muscle tissue (repair) are urgently needed. Beyond the increase in muscle mass, it is important for the new therapies to increase muscle strength and resistance to fatigue. New compounds with these properties (muscle protectors) could counteract long-term space flight muscle atrophy and improve muscle power in aged individuals and in patients with muscle atrophy related to disuse, inflammation or genetic defects.

[0030] Summary of the Invention

[0031] Provided herein is the chemical formula of the new compound 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol or a pharmacologically acceptable isomer, racemate, tautomer, hydrate, isotope or salt thereof.

[0032] The chemical formula can be better explained with reference to Figure 1. In one embodiment, related to 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol compounds are provided having the structure of Formula (II) or a pharmacologically acceptable isomer, racemate, tautomer, hydrate, isotope or salt thereof.

[0033] The chemical formula (II) can be better explained with reference to Figure 2, where A is - 0(R)NH2, where R is lower alkyl or lower alkenyl group

[0034] X1is halogen or haloalkane

[0035] X2is halogen or haloalkane

[0036] R2is lower alkyl or lower alkenyl group

[0037] As used herein, the term “alkyl ' ' means, for example, a branched or unbranched, cyclic or acyclic, Saturated or unsaturated (e.g. alkenyl or alkynyl) hydrocarbyl group which may be substituted or unsubstituted. As used herein, the term “lower alkyl” means, a straight chain or branched alkyl group having from 1 to 8 carbon atoms, in some embodiments from 1 to 6 carbon atoms and in some embodiments from 1 to 4 carbon atoms. Typical lower alkyl groups include, but are not limited to, methyl, ethyl, propyl(n-propyl or isopropyl), butyl (n-butyl, isobutyl or tertiary-butyl), penty and hexyl.

[0038] As used herein, ' 'lower alkenyl ' ' means a straight chain or branched alkenyl group having from 2 to 8 carbon atoms, in some embodiments from 2 to 6 carbon atoms and in some embodiments from 2 to 4 carbon atoms. Alkenyl groups are unsaturated hydrocarbons that contain at least one carbon-carbondouble bond. Examples of lower alkenyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, butenyl, pentenyl and hexenyl.

[0039] As used herein, the term ' 'halogen' ' refers to a fluorine, chlorine, bromine or iodine group, preferably a chlorine or bromine group.

[0040] The term "haloalkane” refers to a lower alkyl as defined herein with one or more hydrogen atoms replaced by halogen. Examples of "haloalkane ” groups include, but are not limited to, -CHF2, CH2F, CFEBr and the like.

[0041] In an embodiment a pharmaceutical composition is provided comprising 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol or a pharmacologically acceptable isomer, racemate, tautomer, hydrate, isotope or salt thereof in combination with a pharmacologically acceptable carrier, diluent or excipient. In an embodiment the pharmaceutical composition is for use in treating microgravity induced damage such as skeletal muscle atrophy, reduced muscle strength or cardiac dysfunction which occur during spaceflight.

[0042] In another embodiment, the pharmacological composition is for use in treating a medical condition associated with muscle atrophy, weakness and reduced endurance capacity under disuse conditions as occurs in orthopaedic patients and in patients in intensive care units. In another embodiment, the pharmacological composition is for use in treating a medical condition associated with muscle atrophy, weakness and reduced endurance capacity such as cancer, heart failure, sepsis, muscle genetic diseases and neurodegenerative disorders.

[0043] In another embodiment, the pharmacological composition is for use in treating a medical condition associated with ageing such as sarcopenia, reduced muscle power and frailty.

[0044] In another embodiment, the pharmacological composition is for use in treating skeletal muscle atrophy and weakness associated with stress induced acceleration in ageing as occurs in diabetic myopathy and sepsis.

[0045] In an embodiment, the pharmacological composition is for use in treating patients with cardiac disease such as patients with heart failure and acute myocardial infarction.

[0046] Useful pharmaceutical carriers for the preparation of the compositions hereof, can be solids, liquids or gases; thus, the compositions can take the form of tablets, pills, capsules, suppositories, powders, enterically coated or other protected formulations (e.g. binding on ionexchange resins or packing in lipid-protein Vesicles), sustained release formulations, solutions, suspensions, elixirs, aerosols, and the like. The compositions maybe subjected to conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, buffers and the like. Such compositions will, in any event, contain an effective amount of the active compound together with a suitable carrier so as to prepare the proper dosage form for proper administration to the recipient. The pharmaceutical preparations can also contain preserving agents, solubilizing agents, stabilizing agents, wetting agents, emulsifying agents, sweetening agents, coloring agents, flavoring agents, salts for varying the osmotic pressure, buffers, coating agents or antioxidants. The compound with the chemical name 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol of the present invention is useful as medicament for the treatment of skeletal muscle atrophy, reduced strength and endurance capacity caused by prolonged microgravity or from any disease resulting in bed confinement and immobilization such as fractures, orthopaedic surgeries, prolonged stay in Intensive Care Units etc. The compound 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol of the present invention is also useful as medicament for the treatment of sarcopenia and frailty caused by ageing and may be useful as medicament for the treatment of muscle atrophy and reduced endurance capacity in common diseases that accelerate ageing, such as cancer, heart failure, sepsis, muscle genetic diseases and inflammatory diseases.

[0047] The therapeutically effective amount or dosage of a compound according to this invention can vary within wide limits and may be determined in a manner known in the art. Such dosage will be adjusted to the individual requirements in each particular case including the specific compound(s) being administered, the route of administration, the condition being treated, as well as the patient being treated. In general, in the case of oral or parenteral administration to adult humans weighing approximately 70 kg, a daily dosage of about 0.03 mg to about 50 mg of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol should be appropriate, although the upper limit may be exceeded when indicated. The dosage is preferably from about 3pg / kg to about 50pg / kg per day. A preferred dosage may be from 5pg / kg to about 20pg / kg per day. The daily dosage can be administered as a single dose or in divided doses, or for parenteral administration it may be given as continuous intravenous infusion.

[0048] In the present invention, we provide strong and unprecedented evidence showing that a novel chemical compound with the chemical Name ”4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol” can treat skeletal muscle wasting, reduced muscle strength and reduced endurance capacity occurring in humans during prolonged exposure to microgravity conditions, as in prolonged space flight and in conditions of immobilization.

[0049] The present invention indicates that administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol during prolonged microgravity conditions retains the ability of myoblasts to form new muscle tissue.

[0050] The present invention indicates that after exposure to prolonged microgravity conditions, administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)- 2, 6-dibromophenoxy)-2 -isopropylphenol upon return to normal gravity recapitulates the ability of myoblasts to form new muscle and the recovery of muscle tissue.

[0051] The present invention is based on the surprising observation that administration of a dosage regimen of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol as considered in the present invention under certain conditions can boost the myogenesis process and result in regeneration of skeletal muscle in vivo or in transplantation of muscle cells or in formation of bioengineered skeletal muscle.

[0052] The present invention in particular concerns a medicament comprising 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol for the treatment of patients with any kind of muscle damage due to unique properties including induction of controlled proliferation and maturation of myoblasts that potentiate the myogenesis process.

[0053] The present invention is based on the surprising observation that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol as considered in the present invention, can effectively orchestrate changes in important kinase signalling, such as p38 MAPK and Akt, in a time-dependent manner to achieve muscle repair and regeneration.

[0054] The present invention is based on the surprising observation that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, can effectively regulate thyroid hormone action at the level of intracellular thyroid hormone receptors to potentiate the myogenesis and achieve muscle repair.

[0055] The present invention is based on the surprising observation that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol as considered in the present invention, can increase SERCA expression and function, regulate calcium homeostasis and achieve muscle repair and functional recovery.

[0056] The present invention is based on the surprising observation that administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, can greatly delay the degeneration of the muscle tissue via inhibition of the processes of senescence and apoptosis.

[0057] The present invention is based on the surprising observation that administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, can effectively treat muscle atrophy as well as reduced muscle power and endurance in patients with prolonged bed confinement and immobilization such as severe trauma patients, acute serious illness and patients in intensive care units.

[0058] The present invention is based on the observation that administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, can effectively treat muscle wasting as well as reduced muscle power and endurance due to cachexia related to inflammation and muscle disuse such as sepsis, diabetes, cancer and heart failure.

[0059] The present invention is based on the observation that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, can effectively treat sarcopenia due to ageing and ageing-accelerating diseases and thus improve resistance to fatigue and frailty in aged individuals.

[0060] The present invention provides strong and unprecedented evidence showing that a novel chemical compound with the chemical Name ”4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenoT' can treat microvascular dysfunction due to increased senescence / apoptosis of endothelial cells occurring in diabetes mellitus and in other human diseases.

[0061] The present invention is based on the observation that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol, improves survival and proliferation of endothelial cells after exposure to high glucose as occurs in diabetes mellitus.

[0062] The present invention concerns a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, that can effectively treat cardiac dysfunction due to heart failure, in particular after myocardial infarction or cardiac dysfunction due to other stresses such as microgravity exposure, critical illness, sepsis and diabetes.

[0063] For example, a specific embodiment is the heart, wherein 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol administration improves contractile force and diastolic function and increases SERCA2a expression an important target protein of new drugs for heart failure. The present invention concerns a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol as considered in the present invention, that can effectively treat cardiac dysfunction induced by long-term exposure to microgravity such as during spaceflights.

[0064] The present invention indicates that administration of a pharmaceutical composition containing 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol can even reduce viability of cancer cells and thus can act as an anti -cancer therapy.

[0065] The invention will now be further described in the Examples below, which are intended as an illustration only and do not limit the scope of the invention.

[0066] Examples

[0067] Example 1. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol induces myoblast maturation, formation of new muscle tissue, inhibits myocyte degeneration and increases cell lifespan

[0068] The effect of the novel compound 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol in the myogenesis process is evaluated in a cell culture model of C2C12 myoblasts. C2C12 are murine skeletal myoblasts derived from satellite cells and their behaviour corresponds to that of progenitor lineage. C2C12 are a subclone of C2 myoblasts, which spontaneously differentiate in culture after serum removal and provide a useful experimental model to study myogenesis and muscle maturation.

[0069] The mouse myoblast C2C12 cells (ATCC #CRL-1772) were cultured at 37 °C under 5% CO2 in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Growth medium). Cells were passaged every 3 days. To induce differentiation, cells that had reached 90% confluence, are switched into differentiation medium consisting of DMEM supplemented with either 1% FBS or 2% serum replacement and 1% penicillin-streptomycin.

[0070] Differentiation medium induces a myogenesis program in C2C12 which includes restricted proliferation, differentiation / maturation of cells and fusion into myotubes to form muscle tissue. During this process, the almost circular myoblast cells become elongated while the actin cytoskeleton becomes denser. The fusion of muscle cells results in development of large polynucleated myotubes. Important molecular markers of the mature skeletal myotubes are the expression of actin and calcium cycling proteins like SERCA1 (Sarcoplasmic Reticulum Calcium ATPase 1). This maturation program is executed within 7-10 days in differentiation medium. At the same time, increased ageing and apoptosis appears in differentiated myotubes especially after day 7 which ultimately leads to degeneration of myotubes after 12-18 days.

[0071] The effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol on proliferation, maturation, fusion and myotube formation as well as apoptosis and senescence are evaluated. The evaluation includes addition of different concentrations of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (4nM, 20nM, lOOnM, 500nM) in the differentiation medium of C2C12 myoblasts. Comparisons are made with a control group including myoblasts cultured in differentiation medium without the compound (vehicle treated) as well as with a group including myoblasts cultured in differentiation medium in the presence of triiodothyronine (T3). T3 is a thyroid hormone that is well known to induce maturation of myoblasts into myotubes. Cultures are observed with an optical inverted microscope Axiovert 25 (Carl Zeiss, Germany) and photographs are taken with a camera Axiocam 208 color (Carl Zeiss, Germany) every 2 days after addition of differentiating medium. The average number of myotubes measured per optical field was used as an index of myogenesis process as reported in the literature.

[0072] In control group, the number of myotubes reaches a peak at day 7 (21±1.5) and then starts to decline indicating the degeneration of the mature myotubes. T3 treatment as expected significantly increased the number of myotubes at day 7 (36±5.8) but still the degeneration process occurs after this point. Figure 3. Surprisingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol administration increased the number of myotubes compared to control but also far beyond the effect of T3 which is a well known maturation factor. In particular, at day 7, the number of myotubes was increased to 50±5 , 75±7.9, 85±4.6 and 87±5.8 after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment with 4nM, 20nM, lOOnM, 500nM, respectively, p<0.05 vs control and T3 group. Beyond any expectations, degeneration was not observed in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol groups after day 7, but the number of myotubes continued to increase till day 9-11. Interestingly, at day 15 there was a great number of mature myotubes in all groups treated with the compound, while no myotubes could be observed in control and T3 groups (Figure 3). To investigate the progress of differentiation process, evaluation of myoblast fusion at day 7 was carried out. Cells were considered fused if they contained two or more nuclei within one cytoplasmic continuity. The cell fusion was assessed by determining the fusion index which was evaluated as the average number of nuclei per myotube. Ten optical fields were randomly chosen. Data were expressed as mean± SD. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at both concentrations of 4nM and 1 OOnM significantly increased the fusion index compared to control group (Figure 4).

[0073] In order to investigate the actin cytoskeleton of the myotubes which is an important index of maturation, C2C12 cells cultured in differentiation medium for 9 days were washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells were incubated with filamentous actin-specific probe phalloidin (Alexa Fluor 488 Phalloidin, #8878, Cell signalling). The nuclei were stained with DAPI (4,6-diamidino-2-phenylindole dihydrochloride; Molecular Probes Inc.) at 0.1 pg / ml in 0.9% NaCl. Cells were examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images were acquired with an integrated camera Axiocam 208 color (Carl Zeiss , Germany). Treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol resulted in more dense actin cytoskeleton. Myotubes after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol are also larger in all diameters.

[0074] Protein extracts of C2C12 cells are obtained by lysing the cells in IX RIPA buffer (10X, #9806, cell signalling). Cell lysates are subjected to electrophoresis by SDS / polyacrylamide gel electrophoresis, separated on 7-10% SDS-polyacrylamide gels and transferred to nitrocellulose membranes (Western Blotting). The membranes are subsequently blocked with 5% non fat milk in Tween-Tris-buffered saline for 1 hour and then incubated with primary antibodies diluted in 5% bovine serum albumin-Tween-Tris-buffered saline (20 mM Tris-HCl, pH 7.4, 150 mM NaCl, and 0.1% Tween 20) (TTBS) overnight at 4°C. Membranes are then washed three times with TTBS buffer (5 min each) and incubated with appropriate anti-mouse (Amersham) or anti-rabbit (Cell Signalling) HRP secondary antibodies. Immunoreactivity is detected by enhanced chemiluminescence using Lumiglo reagents (New England Biolabs). Chemiluminescence is detected by the image analysis system FluorChem HD2 (Alphalnnotech Corporation, 14743, Catalina Street, San Leandro, CA) equipped with a CCD camera and analysis software (Immunoblotting). To detect molecular markers of differentiation such as the sarcoplasmic reticulum calcium ATPase 1 (SERCA1), a protein that is specifically expressed in skeletal muscle cells, an anti-SERCAl primary antibody is used (1:1000; ab2819, Abeam).

[0075] In immature C2C12 myoblasts cultured in growth medium, the expression of SERCA1 is very low. As expected, when myoblasts are cultured in differentiation medium for 3 and 7 days, maturation occurs as indicated by an increase in SERCA1 by around 10 and 20 times respectively. Figure 5. Interestingly, treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol for 3 and 7 days resulted in increased expression of SERCA1 by 2.7 fold and 1.6 fold compared to respective control group, p<0.05. This evidence further supports that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol induces differentiation / maturation of skeletal muscle. Figure 5.

[0076] Example 2. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol orchestrates and potentiates mechanisms of controlled myoblast proliferation and maturation / differentiation involved in myogenesis and repair

[0077] Muscle stem cells can repair / regenerate damaged muscle through a process that includes controlled proliferation, differentiation, fusion, and formation of new muscle tissue. In order to investigate the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on controlled proliferation during this process, C2C12 myoblasts are cultured in differentiation medium for 12 hours and 3 days, are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells are incubated with specific antibody against Ki67 (ab 16667, Abeam ) which is a well known marker of DNA replication. The cell nuclei are also stained with DAPI (4,6-diamidino-2-phenylindole dihydrochloride; Molecular Probes Inc.) at 0.1 pg / ml in 0.9% NaCl. Cells are examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images are acquired with an integrated camera Axiocam 208 color (Carl Zeiss , Germany) and the percentage of positive Ki67 nuclei to total nuclei is evaluated.

[0078] In C2C12 myoblasts cultured in growth medium, the percent of positive Ki67 nuclei reaches 94%±3.7 indicating the high proliferation state as expected. After 12hours in differentiation medium, the percent of positive Ki67 nuclei is already reduced to 61%±9 in control group, p<0.05. Interestingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol treatment results in an increased percent of positive Ki67 nuclei at 90%±3.5 at 12 h. Figure 6. Furthermore, after 3 days in differentiation medium, the percent of positive Ki67 nuclei is greatly reduced to 12.7%±2.6 in control group, whereas in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treated group positive Ki67 nuclei represent 21.9%±4.8, p<0.05. Figure 7. These data indicate that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol results in prolongation of the initial phase of controlled proliferation during myogenesis process.

[0079] Protein extracts from cell lysates cultured in differentiation medium for 12 hours and 3 days were subjected to western blotting and immunoblotting as described above. To investigate mechanisms involved in C2C12 proliferation, we used a primary antibody against Cyclin DI (dilution 1:500, sc-8396, SantaCruz, overnight at 4° C). Cyclins have a central role in cell cycle control during terminal differentiation of muscle cells. Especially, Cyclin DI upregulates in C2C12 by twofold after 9-12 h in differentiation medium followed by a down-regulation to very low levels within 42 h.

[0080] Treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol 4nM resulted in 3.4 fold increased protein content of Cyclin DI after 12 hours in differentiation medium compared to control group, p<0.05. Figure 8. Furthermore, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol 4nM resulted in 2.2 fold increased protein content of Cyclin D 1 after 3 days in differentiation medium compared to control group, p<0.05. Figure 9. These data indicate that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol boosts and prolongs the initial controlled proliferation of myoblasts during the formation of new muscle tissue via cyclin D 1.

[0081] Protein extracts from cell lysates cultured in differentiation medium for 12 hours, 3 days and 7 days, were subjected to western blotting and immunoblotting as described above. To investigate kinase signalling activation which is important in proliferation and differentiation processes, we used primary antibodies against total p38 MAPK and dual phospho-p38 MAPK (Cell Signaling Technology, dilution 1 : 1000), total Akt and dual phospho-Akt (Cell Signaling Technology, dilution 1: 1000), total ERK and dual phospho-ERK (Cell Signaling Technology, dilution 1 : 1000), (Cell Signaling Technology, dilution 1 : 1000) overnight at 4° C. The ratio of phosphorylated to total kinase levels indicating the activation of the kinase is quantified. The protein content of Akt kinase was normalized based on Ponceau staining of the nitrocellulose membrane. After 12 hours in differentiation medium, the ratio of phosphorylated to total p38 MAPK is reduced by 5 fold in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treated myoblasts, p<0.05, while the ratio of phosphorylated to total Akt levels is increased by 1.7 fold in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treated myoblasts compared to control group, p<0.05. No significant changes are found in the ratio of phosphorylated to total p44 MAPK (ERK) in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treated myoblasts. Figure 10.

[0082] After 3 days in differentiation medium, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treatment does not result in significant changes in the activation of p38 MAPK, Akt and p44 MAPK (ERK). However, surprisingly, normalized levels of Akt protein content are increased by 1.75 fold in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treated myoblasts compared to control group, p<0.05. Figure 11.

[0083] After 7 days in differentiation medium, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment does not result in significant changes in the activation of p38 MAPK, Akt and p44 MAPK (ERK). However, surprisingly, normalized levels of Akt protein content are increased by 2.5 fold in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treated myoblasts compared to control group, p<0.05. Figure 12.

[0084] These data indicate that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol orchestrates changes in important kinase signalling, such as p38 MAPK and Akt, in a time-dependent manner to achieve controlled proliferation and enhanced differentiation / maturation which eventually lead to enhanced myogenesis.

[0085] In order to investigate the role of Akt kinase in the effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol in C2C12 myoblasts, we used a specific inhibitor of Akt (GSK-690693). GSK-690693 is known to inhibit all Akt isoforms with an IC50 (inhibitory concentration 50%) of around 2-13 nM. For these experiments, C2C12 myoblasts are cultured in differentiation medium with and without GSK-690693 lOnM in the presence or absence of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (4nM or lOOnM). The average number of myotubes per optical field, indicating the process of myogenesis, is evaluated at different time points. In control group, the number of myotubes per optical field reaches a peak of 24±1.9, while 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment increased the number of myotubes compared to control. In particular, the peak number of myotubes is increased to 64±11 and 76±8 after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment with 4nM and lOOnM, respectively, p<0.05 vs control. Figure 13. Furthermore, addition of GSK results in complete inhibition of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol effects. With GSK, the peak number of myotubes is reduced to 24±3.4 and 29±3 after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment with 4nM and lOOnM, respectively, p<0.05. Figure 13. These data indicate that Akt plays an important role in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol induced myogenesis.

[0086] In order to investigate the potential role of thyroid hormone signalling in the effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol in C2C12 myoblasts, we further used debutyl -dronedarone (DBD), an inhibitor of the transcription factor TRal (thyroid hormone receptor al). DBD is known to inhibit TRal at a concentration of 500nM. For these experiments, C2C12 myoblasts are cultured in differentiation medium with and without DBD 500nM in the presence or absence of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (4nM or lOOnM). The average number of myotubes per optical field, indicating the process of myogenesis, is evaluated at different time points.

[0087] In control group, the number of myotubes per optical field reaches a peak of 23±5 while 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol increased the number of myotubes compared to control. In particular, the peak number of myotubes is increased to 72±10 and 80±10 after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treatment with 4nM and lOOnM, respectively, p<0.05 vs control. Figure 14. Furthermore, administration of DBD results in a dose-dependent inhibition of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol effects. With DBD, the peak number of myotubes is reduced to 11±2.8 and 44±4.7 after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment with 4nM and lOOnM, respectively, p<0.05. Figure 14. These data indicate a potential action of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol via thyroid hormone receptor al. Example 3. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol has anti-apoptotic and anti-ageing properties

[0088] Degeneration of C2C12 cells follows the myogenesis process and is evident especially after 9 days culture in differentiation medium with no FBS. In order to investigate the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on aging and apoptosis, processes that occur during degeneration, C2C12 myoblasts are cultured in differentiation medium for 9 days, are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells are incubated with specific antibody against caspase-3 (#9662, Cell signaling) which is a hallmark of apoptosis. The nuclei are stained with DAPI (4,6-diamidino-2-phenylindole dihydrochloride; Molecular Probes Inc.) at 0.1 pg / ml in 0.9% NaCl. Cells are examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images are acquired with an integrated camera Axiocam 208 color (Carl Zeiss, Germany) and the percentage of positive caspase-3 cells to total number of cells is used as an apoptosis index.

[0089] In another set of experiments, C2C12 myoblasts are cultured in differentiation medium for 9 days, are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and stained with GL13, commercially available as SenTraGor™, to detect the accumulation of lipofuscin within cells which is a hallmark of ageing. The cells are also counterstained with a peroxidase-conjugated polymer backbone - 3,3 '-Diaminobenzidine (DAB) and are examined with a computerized Axioscan Z1 microscope (Carl Zeiss, Germany) using appropriate filter sets. Images are acquired and the percentage of positive lipofuscin cells to total number of cells is used as an senescence index.

[0090] After 9 days in differentiation medium with no FBS, the percent of apoptotic cells is increased to 59%±3 in control group. Interestingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment results in significant inhibition of apoptosis as shown by the low percent of apoptotic cells that reach 13%±2.5 at day 9. p<0.05 vs Control. Figure 15. Furthermore, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol also results in great reduction in the percent of senescent cells after 9 days in differentiation medium (14%±3 in 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol 4nM group as compared to 33%±4 in control group, p<0.05). Figure 16. Thus, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol, beyond any expectations, not only induces the process of myogenesis and the formation of myotubes but also greatly delays the degeneration of the mature myotubes via inhibition of senescence and apoptosis.

[0091] Example 4. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol prevents and repairs damage in cells exposed to simulated microgravity at advanced stages of myogenesis

[0092] Simulated microgravity (s-pg) has been employed on Earth to investigate the effects of real microgravity in space, circumventing the challenges of conducting experiments in space and providing an opportunity to understand the influence of microgravity on living organisms. Simulated microgravity conditions are produced in presented examples using a Random Positioning Machine (RPM). The RPM consists of two rotating frames that are independent of each other; by operating them and changing their direction continuously and randomly, the gravity vector of the earth becomes invalid over time. This device produces an environment similar to that of outer space (10 ’ g) and has been used as a valuable tool in various research fields because it eliminates the influence of gravity and enables controlled arrangements. An RPM experiment consists of a culture chamber mounted on a rotating platform that randomly changes orientation. Constant reorientation creates a continuously changing gravity vector, resulting in a s-pg environment. C2C12 myoblasts were cultured in cell culture flasks for 96h under simulated microgravity at 37 °C. C2C12 myoblasts cultured in the same cell culture flask under normal ground conditions served as a control.

[0093] Important studies demonstrated that exposure to microgravity during spaceflight directly affects skeletal muscle cell differentiation and metabolism. Studies using a RPM to simulate microgravity have observed that exposure to s-pg in C2C12 and L6 cells blocks myogenesis by decreasing myosin expression, myotube thickness, and myotube fusion index. These results suggest that simulated microgravity causes similar phenotypic changes in skeletal muscle as observed during spaceflight.

[0094] In order to investigate the effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol on microgravity induced damage in advanced stages of myogenesis, we culture C2C12 myoblasts in differentiation medium (1% FBS) under normal ground conditions for 6 days, we expose the cells in simulated microgravity for 96 hours (from day 6 to day 10) and we return the cells to normal gravity after day 10 (recovery period). 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol is added in culture at a concentration of lOOnM either at day 6 (early) or at day 10 (late) to simulate different treatment options either during spaceflight or upon return to earth. The average number of myotubes per optical field, indicating the process of myogenesis, is evaluated at different time points.

[0095] At the developmental stage of day 6 of myogenesis, the average number of mature myotubes that has already been formed is 16.5±2.2 per optical field. Exposure of cells to simulated microgravity (s-pg) for 96 hours at this stage not only stops further maturation and formation of myotubes but also leads to degeneration resulting in significantly reduced average number of myotubes of 6.4±3.5 at day 10. Figure 17. At the same time, exposure to s-pg causes cell detachment which is evident as empty patches in the cellular layer at day 10. Furthermore, even after return to normal gravity conditions, cells only partially recover their ability for myotube formation resulting in an average number of myotubes of 29±4 at day 16 in s-pg group compared to 92±1.7 in Control group. Surprisingly, treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol lOOnM during s-pg exposure, retains the ability of cells to form myotubes and increases the average number of myotubes to 27±2.4 in s-pg+CX(early) group at day 10, p<0.05 vs s-pg group. Figure 17. Beyond any expectations, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment stops s-pg induced cell detachment and empty patches are not observed in the cellular layer. During recovery period, myogenesis is further potentiated in s-pg+CX(early) group and the detrimental effect of s-pg almost disappears resulting in an average number of myotubes of 75±11 at day 16, p<0.05 vs s-pg . Figure 17. When 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol was added after exposure to s-pg, at day 10, recovery of myogenesis has been potentiated resulting in increased number of myotubes but still the effect of s-pg is evident. In fact the average number of myotubes is 53±4 at day 16 in s-pg+CX(late) group as compared to 23±4 in s-pg, p<0.05. Figure 18.

[0096] Example 5. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol prevents and repairs damage in cells exposed to simulated microgravity at early stages of myogenesis

[0097] Simulated microgravity conditions are produced in presented examples using a Random Positioning Machine (RPM) as described above producing an environment similar to that of outer space (10 ’ g). In order to investigate the effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol on microgravity induced damage in the initial stage of myogenesis, we culture C2C12 myoblasts in differentiation medium (1% FBS) under normal ground conditions for 2 days, we expose the cells in simulated microgravity for 96 hours (from day 2 to day 6) and we return the cells to normal gravity after day 6 (recovery period). 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol is added in culture at a concentration of lOOnM either at day 2 (early) or at day 6 (late) to simulate different treatment options either during spaceflight or upon return to earth. The average number of myotubes per optical field, indicating the process of myogenesis, is evaluated at different time points. At the developmental stage of day 2 of myogenesis, very few myotubes have been formed. Exposure of cells to simulated microgravity (s-pg) for 96 hours at this stage stops maturation and formation of myotubes resulting in an average number of myotubes of 4.1±1.4 per optical field at day 6. It is of interest that at this stage, exposure to s-pg does not cause cell death and no empty patches in the cellular layer are evident at day 6. Furthermore, after return to normal gravity conditions, cells totally recover their ability for myotube formation resulting in an average number of myotubes of 59±3 at day 10. Figure 19. However, exposure to s-pg results in acceleration of the degeneration process after day 12 resulting in an average number of myotubes of41±12 per optical field at day 16 in s-pg group vs 92±1.7 in Control group, p<0.05. Surprisingly, treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol lOOnM during exposure to simulated microgravity, not only retains the ability of cells to form myotubes and increases the average number of myotubes to 28±3 in s-pg+CX(early) group at day 6, p<0.05 vs s-pg group, but also results in significant morphological changes. Figure 19. Beyond any expectations, the combination of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and s-pg results in larger and more robust myotubes at day 6. During recovery period, myogenesis is further potentiated in s-pg+CX(early) group and 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol abrogates the acceleration of myotube degeneration caused by s-pg resulting in an average number of myotubes of 80±2 per optical field at day 16 not significantly different from control group. Figure 19. When 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol was added after exposure to s-pg, at day 6, recovery of myogenesis has been potentiated resulting in increased number of myotubes but still the effect of s-pg is evident. Figure 20. Example 6. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol inhibits the detrimental effect of simulated microgravity to myoblast proliferation Simulated microgravity conditions are produced in presented examples using a Random Positioning Machine (RPM) as described above producing an environment similar to that of outer space (10 ’ g). In order to investigate the effects of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on microgravity induced inhibition of myoblast proliferation, we culture C2C12 myoblasts in growth medium (10%FBS) under normal ground conditions for 24 hours and then we expose the cells in simulated microgravity for 96 hours while we keep them in growth medium (s-pg group). 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol is added in culture at a concentration of lOOnM at initiation of exposure to simulated microgravity (s-pg+CX group). A control group is also examined with C2C12 myoblasts cultured in growth medium under normal ground conditions for 96 hours. After 96 hours, cells are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells are incubated with specific antibody against Ki67 (ab 16667, Abeam ) which is a well known marker of DNA replication and nuclei are also stained with DAPI as described above. Cells are examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images are acquired with an integrated camera Axiocam 208 color (Carl Zeiss , Germany) and the percentage of positive Ki67 nuclei to total nuclei is evaluated.

[0098] In C2C12 myoblasts cultured in growth medium under normal gravity for 96 hours, the percent of positive Ki67 nuclei reaches 59%±4 indicating a high proliferation state as expected. However, after simulated microgravity for 96hours in growth medium, the percent of positive Ki67 nuclei is dramatically reduced to 2%±2 in s-pg group, p<0.05. Surprisingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treatment results in an increased percent of positive Ki67 nuclei at 69%±5 after 96 hours in simulated microgravity. Figure 21. These data indicate that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol abrogates the detrimental effect of microgravity in myoblast proliferation which is directly linked with the regenerative capacity of the injured muscle.

[0099] Example 7. Administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol inhibits muscle atrophy, increases muscle strength and resistance to fatigue after hindlimb immobilization Muscle atrophy was induced in C56 / B16 mice via leg casting. After induction of anaesthesia with ketamine and midazolam, the left hindlimb was maintained in a knee joint extension and ankle plantar extension position and taped with a non-elastic bandage tape to prevent edema and dermatitis. The left hindlimb was fixed by a 12-15 mm -wide cast in all animals. Casting was removed after 7 days. This model of immobilization has been shown to result in significant atrophy of gastrocnemius muscle, simulates well clinical conditions such as limb immobilization and atrophy after fractures or orthopedic surgeries in humans and is considered the appropriate experimental model for the study of conditions of immobilization and muscle disuse such as microgravity, prolonged bed confinement and ageing. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol was administered in two groups at two different doses of 70 pg / Kg and 400 pg / Kg intraperitoneally once a day starting a few hours after casting and extended till the end of the experiment. Animals in control group were treated with normal saline. Animals were studied at 2 different time-points: 24 hours and 8 days after removal of casting. Measurements included weight of the left (immobilized) and right gastrocnemius muscle. The percentage of gastrocnemius muscle weight loss was calculated as the difference between the weight of the right and left gastrocnemius to the weight of the right gastrocnemius. Unilateral muscle strength of the left hindlimb was measured in grams using a modified grip strength meter (BioSeb). We also measured endurance capacity using a Treadmill (Panlab) to perform a fatigue test. This treadmill includes a moving belt with regulated speed and an electric metal grid located at the rear of the moving belt as an external motivator. The intensity of the electric shock is adjusted at 1.2mA to produce no more than a mild tingling sensation when touched. The protocol includes regulation of the belt speed at lOcm / sec for Imin, 20cm / sec for 2min, 25cm / sec for 5min, 30cm / sec for 15min, 35cm / sec for 15min, 40cm / sec for 15min and 45cm / sec for 15min. When the mice fatigue they spend more time toward the back of the treadmill. The criterion for exhaustion in this protocol is defined as receiving more than five shocks in less than 60 seconds. The total distance (in meters) travelled till reaching exhaustion and the duration of the test is recorded.

[0100] As expected, leg casting for 7 days resulted in significant muscle atrophy. At 24 hours after removal of casting, the percentage of gastrocnemius muscle weight loss for the left hindlimb compared to the right was 26%±6.1 in control group. Interestingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 70 pg / Kg and 400 pg / Kg, resulted only in 12.5%±5.6 and 12.9%±0.6 gastrocnemius muscle weight loss respectively (p<0.05 vs control for both). Figure 22. Furthermore, the grip strength of the left hindlimb was found to be reduced at 42.8±8.7 grams in control group, 24 hours after removal of casting. Interestingly, administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 70 pg / Kg and 400 pg / Kg, resulted in 68±11 grams and 69±10 grams grip strength respectively (p<0.05 vs control for both). Figure 23. The total distance measured by the treadmill fatigue test was also found to be reduced at 163±113 m in control group at 24 hours. Administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 70 pg / Kg and 400 pg / Kg, resulted in significantly increased total distance at 279±149m and 243±148m respectively (p<0.05 vs control for both). Figure 24.

[0101] At day 8 after removal of casting, the grip strength of the left hindlimb was found to be partially increased in control group at 66±12 grams compared to 24 hours. However, the grip strength was further increased in animals after administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 70 pg / Kg and 400 pg / Kg, resulting in 80±10 grams and 81±8 grams grip strength respectively (p<0.05 vs control for both). Figure 25. The total distance measured by the treadmill fatigue test was found to be at 288±37m in control group at day 8. It is of note that administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 70 pg / Kg and 400 pg / Kg, resulted in a great increase in total distance at 446±65m and 728±76m respectively (p<0.05 vs control for both). Figure 26.

[0102] These results indicate that the compound 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol counteracts muscle atrophy and weakness caused by immobilization and significantly increases endurance capacity.

[0103] Example 8. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol increases proliferation and inhibits apoptosis and cellular senescence induced by high glucose in human microvascular endothelial cells

[0104] The effect of the new compound 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol in the high glucose induced injury was tested in a model of cultured human microvascular endothelial cells. High glucose administration is a useful experimental model for simulating the deleterious effects of diabetes mellitus in cell culture models. Microvascular endothelial cells determine microvascular function and therefore the blood supply to various organs. Dysfunction and rarefaction of small vessels occurs in both diabetes mellitus and in muscle tissue after immobilization and disuse. Microvascular dysfunction is recognized as a central mechanism in the pathogenesis of sepsis as well as in ICU-induced muscle weakness. Inflammatory activation of the endothelium in severe disease results in the secretion of numerous mediators that can affect muscle function. Thus, the damage of skeletal muscle through endothelial-muscle cell interaction is a central theory for explaining ICU-induced muscle weakness. [Front. Physiol. 2023, 14:1170429],

[0105] The human microvascular endothelial cells were obtained from Innoprot (P10880 Innoprot, Spain) and cultured at 37 °C under 5% CO2 in Petri dishes or flasks coated with fibronectin in culture medium using EMEM (P60104 Innoprot, Spain) supplemented with 5% fetal bovine serum (FBS) and 1% penicillin-streptomycin (Growth medium) till 90-95% confluence. Medium was changed every 3 days. Cells were subjected to high glucose stress (32mM) for 48 hours while in another group 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol was administered at a dose of lOOnM with high glucose. High glucose stress is known to reduce proliferation and increase apoptosis and senescence in endothelial cells.

[0106] In order to investigate the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol on proliferation during this process, endothelial cells are are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells are incubated with specific antibody against Ki67 (ab 16667, Abeam) which is a well known marker of DNA replication. The cell nuclei are also stained with DAPI (4,6-diamidino-2-phenylindole dihydrochloride; Molecular Probes Inc.) at 0.1 pg / ml in 0.9% NaCl. Cells are examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images are acquired with an integrated camera Axiocam 208 color (Carl Zeiss , Germany) and the percentage of positive Ki67 nuclei to total nuclei is evaluated.

[0107] In endothelial cells cultured with normal glucose concentration (control group), the percent of positive Ki67 nuclei reaches 44%±4.8 indicating the high proliferation capacity as expected. After 48 hours in high glucose concentration, the percent of positive Ki67 nuclei is dramatically reduced to 7.6%±2.0 in HG group, p<0.05 compared to control. Interestingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treatment results in an increased percent of positive Ki67 nuclei at 16.3%±2.4 in HG+CX group, p<0.05 compared to HG group. Figure 27. In order to investigate the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on cellular senescence and apoptosis, processes that occur after exposure to high glucose, endothelial cells are washed in phosphate-buffered saline (PBS), fixed in PBS-4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Fixed cells are incubated with specific antibody against caspase-3 (#9662, Cell signaling) which is a hallmark of apoptosis or with specific antibody against p21 factor (Anti-p21 antibody, abeam, ab227443) which is implicated in mechanisms of cellular senescence. Secondary fluorescent antibodies against the primary antibodies were used in order to detect the signal of apoptosis or senescence. The nuclei are stained with DAPI (4,6-diamidino-2-phenylindole dihydrochloride; Molecular Probes Inc.) at 0.1 pg / ml in 0.9% NaCl. Cells are examined with an epifluorescence Axiovert 100 light microscope (Carl Zeiss, Germany), using appropriate filter sets. Images are acquired with an integrated camera Axiocam 208 color (Carl Zeiss, Germany) and the percentage of positive caspase-3 or p21 cells to total number of cells is used as an apoptosis or senescence index, respectively.

[0108] After 48 hours in high glucose, the percent of apoptotic cells is increased to 12.5%±4.4 in HG group compared to 3.4%±1.3 in control group. Interestingly, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment results in significant inhibition of apoptosis as shown by the low percent of apoptotic cells that reach 4.3%±1.1 in group HG+CX, p<0.05 vs HG. Figure 28. Furthermore, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol also results in great reduction in the percent of senescent cells after 48 hours in high glucose (10%±3.2 in group HG+CX as compared to 35.6%±7.9 in HG group, p<0.05). Figure 29.

[0109] Thus, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol, beyond any expectations, restores the impaired proliferation capacity of endothelial cells and significantly inhibits senescence and apoptosis induced by high glucose. The integrity of the endothelium is related to improvement of microvascular dysfunction, improved tissue perfusion and enhanced functional capacity of skeletal muscles and other organs in diabetes.

[0110] Example 9. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol regulates the expression of myocardial contractile proteins and potentiates cardiac function The effects of novel 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol compound in cardiac function are determined in healthy animals in vivo. In Wistar rats, three different doses of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol 5 pg / Kg, 20 pg / Kg and 100 pg / Kg are injected intraperitoneally once a day for a total of 2 weeks. No mortality or toxic effects are observed after administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol in vivo for 2 weeks. The study investigated the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on heart rate, global contractile force of the left ventricle of the heart (ejection fraction), regional contractile force of the left ventricular wall of the heart (Systolic velocity of LV posterior wall) and relaxation of left ventricular wall of the heart (Diastolic velocity of LV posterior wall) using echocardiography. Furthermore, the levels of expression of important calcium handling proteins of the heart are studied in protein extracts of left ventricular tissue by western blotting. In particular, the protein content of Sarcoplasmic Reticulum calcium ATPase (SERCA2a) and phospholamban is determined. Reduction of SERCA2a and / or increase of phospholamban lead to severe impairment of calcium handling and contractile dysfunction serving as a hallmark of heart failure.

[0111] No changes are observed in heart rate after 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol treatment with different doses. Figure 30. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at the dose of 100 pg / Kg significantly increased global contractile force of the heart and regional contractile force of the left ventricular wall. Figure 31 and 32. Furthermore, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at a dose of 100 pg / Kg significantly increased relaxation of left ventricular wall of the heart. Figure 33. Quantitative assessment of calcium handling proteins in heart tissue of rats reveals that 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at a dose of 100 pg / Kg for 2 weeks increases Sarcoplasmic Reticulum calcium ATPase (SERCA2a) to phospholamban protein expression compared to Control group. Figure 34. The above data indicate important unique properties of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol compound of the invention to enhance cardiac contractile function (positive inotropic effect). These properties could be used for the treatment of acute or chronic heart failure. Example 10. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol improves cardiac function after ischemia-reperfusion

[0112] Wistar male rats are used for this study. The isolated rat heart model is used to simulate myocardial ischemia-reperfusion ex vivo. Rat hearts are perfused with oxygenated Krebs buffer containing electrolytes and glucose under normal temperature. A balloon inside the left ventricle of the heart allows measurement of left ventricular developed pressure (LVDP) which is an index of contractile force of the heart and left ventricular end-diastolic pressure (LVEDP) which indicates the diastolic function of the heart. This model simulates the clinical conditions of myocardial infarction caused by coronary vessel occlusion (ischemia) and reopening of the vessel with percutaneous coronary intervention (reperfusion) in patients. In the isolated rat heart model, ischemia is caused by stopping the perfusion with the oxygenated Krebs buffer for 30 min and reperfusion follows by reinstatement of the perfusion with the buffer for 60 min. Ischemia results in myocardial damage leading to reduced contractile force of the heart (LVDP) and increased LVEDP during reperfusion that represents diastolic dysfunction. It should be noted that such an increase in LVEDP may lead to pulmonary edema the clinical setting, while reduction in contractile force results in heart failure in patients. Furthermore, the reduction in contractile force of the heart is expressed as percentage of the initial LVDP (recovery of LVDP) in this model.

[0113] In particular, isolated control hearts are subjected to 20 min of normal perfusion, 30 min of zero-flow ischemia and 60 min of reperfusion with addition of vehicle in the perfusate 10 min after reperfusion (Control group, n= 11). Another group of hearts are subjected to 20 min of normal perfusion, 30 min of zero-flow ischemia and 60 min of reperfusion with addition of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol 4nM in the perfusate 10 min after reperfusion (CX group, n=10).

[0114] After normal perfusion and before ischemia, myocardial function is similar in both groups. Thus, LVDP and LVEDP is 129±14 mmHg and 8.0±0.5mmHg in control group versus 125±18mmHg and 8.1±0.4 mmHg in CX group, respectively. Interestingly, treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at reperfusion improves diastolic dysfunction, reduced LVEDP and improved force of contraction. After 60min of reperfusion, LVEDP is 63±9 mmHg in Control group and 53±11 mmHg in CX group, p<0.05. Figure 35. Most importantly, recovery of myocardial function is increased with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol treatment indicating that this new compound improves cardiac dysfunction after myocardial infarction. In particular, after 60min of reperfusion, recovery of LVDP is 36±11% in Control group and 47±10% in CX group, p<0.05. Figure 36. The above data indicate important unique properties of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol compound of the invention to enhance myocardial recovery of function under clinical conditions of ischemia-reperfusion, such as acute myocardial infarction, coronary angioplasty, donor heart preservation during transplantation, cardiac surgery etc.

[0115] Example 11. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol does not increase or even reduces viability of cancer cell lines

[0116] To test the effect of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on cancer cells, we use 2 cancer cell lines; the human estrogen receptor positive breast adenocarcinoma adherent cell line, MCF-7, and the colon cancer cells. Cells were obtained from either the American Type Culture Collection (ATCC) or American Type European Collection of Authenticated Cell Cultures (ECACC).

[0117] The MCF-7 cells are maintained in Dulbecco's Modified Eagles' Medium (DMEM) supplemented with 10% fetal bovine serum, 100 U mb1penicillin-streptomycin. The cells are cultured in humidified incubator supplied with 5% CO2 and maintained at 37 °C.

[0118] Colon cancer cells are maintained in Roswell Park Memorial Institute (RPMI) Medium supplemented with 10% fetal bovine serum, 100 U mb1penicillin-streptomycin. The cells are cultured in humidified incubator supplied with 5% CO2 and maintained at 37 °C.

[0119] The impact of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol on cancer cell viability was evaluated by performing 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Vybrant® MTT Cell Proliferation Assay kit, V-13154). The MTT assay is a colorimetric method that involves measuring the reduction of yellow 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) by mitochondrial succinate dehydrogenase. The principle is based on the number of cells present and the assumption that those dead cells or their products do not reduce tetrazolium. The MTT enters into the mitochondria in the cells and is reduced to purple-colored formazan crystals, which are insoluble. The cells are treated with MTT (500 pg / mL) for 4 h. Dimethyl sulfoxide (DMSO) (10%) is added to dissolve the formazan formed crystals. Finally, cell viability at an optical density (OD) of 570 nm is measured using a Multiscan™ FC Microplate Photometer (Thermo Scientific, USA). 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol is added at concentrations of 4nM and lOOnM.

[0120] The administration of 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol lOOnM in MCF-7 cancer cell culture resulted in a statistically significant reduction in cell survival on both the 5th and 9th day. Figure 37. In the case of colorectal cancer cells, 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol administration at both 4nM and lOOnM doses led to a statistically significant reduction in cell survival on the 5th day, but there was no difference on the 7th and 9th days. Figure 38.

[0121] Brief Description of the Figures

[0122] The present invention will now be described in detail with reference to accompanying drawings:

[0123] • Figure 1 shows the new compound with the Chemical Name 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol, Molecular Formula: CisiffeB NCh, Molecular Weight: 473.21

[0124] • Figure 2 shows the chemical structure of formula (II) describing related to 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol compounds

[0125] • Figure 3. Average number of myotubes measured at different time-points after culture of C2C12 myoblasts in differentiation medium (no FBS). The graph shows the progression of the myogenesis process and the effect of 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol (CX) at different concentrations compared to a well-known maturation factor such as triiodothyronine (T3). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control and T3

[0126] • Figure 4. The graph shows the evaluation of myoblast fusion process after 7 days in differentiation medium (no FBS). The fusion index represents the average number of nuclei per myotube as assessed by optical microscopy photos. Ten optical fields were randomly chosen. Data were expressed as mean ± SD. 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol (CX) at both concentrations of 4nM and lOOnM significantly increased the fusion index compared to control group.

[0127] • Figure 5. SERCA1 content, a molecular index of muscle maturation, is significantly increased after treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol (CX). *p<0.05 vs Control

[0128] • Figure 6. Percent of Ki67 positive nuclei representing DNA replication of C2C12 cells cultured in growth medium (myoblasts) and after 12 hours in differentiation medium (Control and 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol 4nM). Differentiation medium results in decreased Ki67 as early as 12 hours while treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol (CX) prolongs the DNA replication process. * p<0.05 vs Myoblasts, **p<0.05 vs Control.

[0129] • Figure 7. Percent of Ki67 positive nuclei representing DNA replication of C2C12 cells cultured in differentiation medium for 3 days (Control and 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol 4nM). Treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (CX) prolongs the DNA replication process. **p<0.05 vs Control.

[0130] • Figure 8. Quantification of protein content of Cyclin DI, a well known marker of proliferation, in C2C12 cells cultured in differentiation medium for 12 hours in the presence (CX) or absence (Control) of 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol. Treatment with 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol (CX) results in increased Cyclin DI expression. * p<0.05 vs Control.

[0131] • Figure 9. Quantification of protein content of Cyclin DI, a well known marker of proliferation, in C2C12 cells cultured in differentiation medium for 3 days in the presence (CX) or absence (Control) of 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol. Treatment with 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol results in increased Cyclin DI expression. * p<0.05 vs Control.

[0132] • Figure 10. The graphs show the activation of p38 MAPK, Akt and p44 MAPK after 12 hours culture of C2C12 cells in differentiation medium in Control and 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (CX) treated groups.

[0133] 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol inhibits activation of p38 MAPK and induces activation of Akt. *p<0.05 vs Control

[0134] • Figure 11. The graphs show the activation of p38 MAPK, Akt and p44 MAPK after 3 days culture of C2C12 cells in differentiation medium in Control and 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (CX) treated groups. In addition, the protein content of Akt after 3 days culture of C2C12 cells in differentiation medium in Control and 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2-isopropylphenol (CX) treated groups is shown. 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol increases Akt expression after 3 days. *p<0.05 vs Control

[0135] • Figure 12. The graphs show the activation of p38 MAPK, Akt and p44 MAPK after 7 days culture of C2C12 cells in differentiation medium in Control and 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (CX) treated groups. In addition the protein content of Akt after 7 days culture of C2C12 cells in differentiation medium in Control and 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2-isopropylphenol (CX) treated groups is shown. 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol (CX) increases Akt expression after 7 days. *p<0.05 vs Control

[0136] • Figure 13. Average number of myotubes measured at different time-points after culture of C2C12 myoblasts in differentiation medium (no FBS). The graphs show the progression of the myogenesis process in the presence (CX) or absence (Control) of 4- (4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and the effect of a well known inhibitor of Akt (GSK-690693). 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2-isopropylphenol (CX) is used at a low (4nM, A) and high concentration (lOOnM, B). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control, GSK and CX+GSK

[0137] • Figure 14. Average number of myotubes measured at different time-points after culture of C2C12 myoblasts in differentiation medium (no FBS). The graphs show the progression of the myogenesis process in the presence (CX) or absence (Control) of 4- (4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol and the effect of a well known inhibitor of thyroid hormone receptor al (Debutyl-dronedarone, DBD). 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (CX) is used at a low (4nM, A) and high concentration (lOOnM, B). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control, DBD and CX+DBD, **p<0.05 vs Control

[0138] • Figure 15 shows: the quantification of apoptosis expressed as the percent of cells positive for caspase-3 to total cells in the presence (CX) or absence (Control) of 4-(4- ((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol after culture for 9 days in differentiation medium. 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)- 2-isopropylphenol significantly reduces apoptosis. *p<0.05 vs Control

[0139] • Figure 16 shows: the quantification of cellular senescence expressed as the percent of cells positive for lipofuscin staining to total C2C12 cells cultured in differentiation medium for 9 days with (CX 4nM) and without (Control) 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol. 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol (CX) significantly reduces cellular senescence. *p<0.05 vs Control

[0140] • Figure 17 shows average number of myotubes (myogenesis process) measured at different time-points after culture of C2C12 myoblasts in differentiation medium (1% FBS) in Control group (normal gravity conditions) and after exposure to simulated microgravity from day 6 to day 10 with 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol [s-pg+CX(early) group] and without [s-pg] . 4-(4- ((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol 1 OOnM was added from day 6 to day 20 (grey bar). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control and s-pg+CX(early), **p<0.05 vs Control • Figure 18 shows average number of myotubes (myogenesis process) measured at different time-points after culture of C2C12 myoblasts in differentiation medium (1% FBS) in Control group (normal gravity conditions) and after exposure to simulated microgravity from day 6 to day 10 with 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol [s-pg+CX(late) group] and without [s-pg] . 4-(4- ((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol lOOnM was added from day 10 to day 20 (grey bar). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control and s-pg+CX(late), **p<0.05 vs Control

[0141] • Figure 19 shows average number of myotubes (myogenesis process) measured at different time-points after culture of C2C12 myoblasts in differentiation medium (1% FBS) in Control group (normal gravity conditions) and after exposure to simulated microgravity from day 2 to day 6 with 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol [ s-pg+CX(carl ) group] and without [s-pg] . 4-(4- ((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol lOOnM was added from day 2 to day 20 (grey bar). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control and s-pg+CX(early), **p<0.05 vs Control

[0142] • Figure 20 shows average number of myotubes (myogenesis process) measured at different time-points after culture of C2C12 myoblasts in differentiation medium (1% FBS) in Control group (normal gravity conditions) and after exposure to simulated microgravity from day 2 to day 6 with 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol [s-pg+CX(late) group] and without [s-pg]. 4-(4- ((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol lOOnM was added from day 6 to day 20 (grey bar). Data are expressed as Mean with Standard Deviation. * p<0.05 vs Control and s-pg+CX(late), **p<0.05 vs Control

[0143] • Figure 21. Percent of Ki67 positive nuclei representing DNA proliferation in C2C12 myoblasts cultured in growth medium for 96 hours in normal ground conditions (Control) and in C2C12 myoblasts cultured in growth medium for 96 hours in microgravity conditions with (CX) and without 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2-isopropylphenol. S-pg dramatically reduces proliferation rate of myoblasts while 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol abrogates this effect. * p<0.05 vs Control, ** p<0.05 vs s-pg

[0144] • Figure 22 illustrates percentage of gastrocnemius muscle weight loss of the left hindlimb compared to the right, 24 hours after removal of casting in control group and in groups receiving 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol at doses of 70 pg / Kg and 400 pg / Kg [CX(70) and CX(400)]. *p<0.05 vs Control

[0145] • Figure 23 illustrates the grip strength of the left hindlimb 24 hours after removal of casting in control group and in groups receiving 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol at doses of 70 pg / Kg and 400 pg / Kg [CX(70) and CX(400)]. *p<0.05 vs Control

[0146] • Figure 24 illustrates the total distance measured by the treadmill fatigue test, 24 hours after removal of casting in control group and in groups receiving 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at doses of 70 pg / Kg and 400 pg / Kg [CX(70) and CX(400)]. *p<0.05 vs Control

[0147] • Figure 25 illustrates the grip strength of the left hindlimb, 8 days after removal of casting in control group and in groups receiving 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol at doses of 70 pg / Kg and 400 pg / Kg [CX(70) and CX(400)]. *p<0.05 vs Control

[0148] • Figure 26 illustrates the total distance measured by the treadmill fatigue test, 8 days after removal of casting in control group and in groups receiving 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at doses of 70 pg / Kg and 400 pg / Kg [CX(70) and CX(400)]. *p<0.05 vs Control

[0149] • Figure 27 illustrates the percent of Ki67 positive nuclei representing DNA replication of microvascular endothelial cells cultured in normal glucose (Control) and in high glucose conditions in the absence (HG) or presence of 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol lOOnM (HG+CX). Treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (CX) promotes proliferation of endothelial cells. *p<0.05 vs Control, **p<0.05 vs HG

[0150] • Figure 28 illustrates quantification of apoptosis expressed as the percent of endothelial cells positive for caspase-3 to total cells when cultured in normal glucose (Control) and in high glucose conditions in the absence (HG) or presence of 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol lOOnM (HG+CX). Treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (CX) significantly reduces apoptosis of endothelial cells induced by high glucose. *p<0.05 vs Control, **p<0.05 vs HG

[0151] • Figure 29 illustrates cellular senescence expressed as the percent of endothelial cells positive for p21 to total cells when cultured in normal glucose (Control) and in high glucose conditions in the absence (HG) or presence of 4-(4-((4-Aminobutan-2yl)oxy)- 2,6-dibromophenoxy)-2-isopropylphenol lOOnM (HG+CX). Treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (CX) significantly reduces senescence of endothelial cells induced by high glucose. *p<0.05 vs Control, **p<0.05 vs HG

[0152] • Figure 30 illustrates heart rate, in healthy rats after 2 weeks of treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol compound (CX) of the invention at doses of 5 pg / Kg, 20 pg / Kg and 100 pg / Kg.

[0153] • Figure 31 illustrates global contractile force of the heart (ejection fraction), in healthy rats after 2 weeks of treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol compound (CX) of the invention at doses of 5 pg / Kg, 20 pg / Kg and 100 pg / Kg. 4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2 -isopropylphenol at 100 pg / Kg increases global contractile force of the heart compared to vehicle treated Control group. *p<0.05 vs Control

[0154] • Figure 32 illustrates regional contractile force of the myocardial wall (Systolic velocity of LV posterior wall), in healthy rats after 2 weeks of treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol compound (CX) of the invention at doses of 5 pg / Kg, 20 pg / Kg and 100 pg / Kg. 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at 100 pg / Kg increases regional contractile force of the myocardial wall compared to vehicle treated Control group. *p<0.05 vs Control

[0155] • Figure 33 illustrates relaxation of left ventricular wall of the heart (Diastolic velocity of LV posterior wall) in healthy rats after 2 weeks of treatment with 4-(4-((4- Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol compound (CX) of the invention at doses of 5 pg / Kg, 20 pg / Kg and 100 pg / Kg. 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at 100 pg / Kg increases relaxation of left ventricular wall of the heart compared to vehicle treated Control group. *p<0.05 vs Control

[0156] • Figure 34 illustrates quantitative assessment of Sarcoplasmic Reticulum calcium ATPase (SERCA2a) to phospholamban protein content in heart tissue of healthy rats after 2 weeks of treatment with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)- 2-isopropylphenol compound (CX) of the invention at doses 5 pg / Kg, 20 pg / Kg and 100 pg / Kg.4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol at 100 pg / Kg increases the ratio of SERCA2 -phospholamban compared to vehicle treated Control group. *p<0.05 vs Control

[0157] • Figure 35 shows the left ventricular end-diastolic pressure (LVEDP) indicating diastolic dysfunction after 30 min of ischemia and 60 min of reperfusion in vehicle treated (Control) and 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2- isopropylphenol (CX) treated hearts. *p<0.05 vs Control

[0158] • Figure 36 shows the recovery of left ventricular developed pressure (LVDP) indicating contractile force after 30 min of ischemia and 60 min of reperfusion in vehicle treated (Control) and 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol (CX) treated hearts. *p<0.05 vs Control

[0159] • Figure 37 illustrates cell viability of MCF-7 cells cultured without (Control) or with 4- (4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at concentration of 4nM and lOOnM (CX 4nM and CX lOOnM). *p<0.05 vs Control and CX 4nM, **p<0.05 vs Control

[0160] • Figure 38 illustrates cell viability of colon cancer cells cultured without (Control) or with 4-(4-((4-Aminobutan-2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol at concentration of 4nM and lOOnM (CX 4nM and CX lOOnM). **p<0.05 vs Control

Claims

CLAIMS1. A new compound with the chemical name ' '4-(4-((4-Aminobutan-2yl)oxy)-2,6- dibromophenoxy)-2-isopropylphenol” of the formula shown hereor a pharmacologically acceptable isomer, racemic mixture, tautomer, isotope, hydrate or salt thereof.

2. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemic mixture, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of muscle atrophy, muscle weakness and reduced resistance to fatigue from any cause.

3. Pharmaceutical composition according to claims 1 and 2 for use in the treatment of skeletal muscle injury caused by exposure to microgravity and leading to muscle atrophy, muscle weakness and reduced resistance to fatigue.

4. Pharmaceutical composition according to claim 3 for use in the treatment of skeletal muscle injury caused by exposure to microgravity conditions, in case of preventive administration.

5. Pharmaceutical composition according to claim 3 for use in the treatment of skeletal muscle injury caused by exposure to microgravity conditions, in case of administration during return to normal gravity.

6. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claims 1 and 2or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in ageing-related sarcopenia, musculoskeletal trauma, diabetic myopathy and muscle atrophy in Intensive Care Units for the treatment of reduced regenerative capacity of skeletal muscles, in cases when satellite muscle cells fail to proliferate, mature and create muscle tissue 7. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol according to claims 1 and 2 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of muscle mass loss, muscular weakness and reduced resistance to fatigue due to long-term immobilization and / or bed confinement, e.g. in patients with fractures, orthopedic surgeries, severe trauma and patients in intensive care units.

8. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol according to claims 1 and 2 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of cachexia, muscle mass loss and atrophy due to inflammation and muscle disuse as occurs in sepsis, diabetes, cancer and heart failure.

9. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol according to claims 1 and 2 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of sarcopenia, general weakness and reduced strength due to ageing and ageing -accelerating diseases.

10. Pharmaceutical composition according to claim 9, for use in the treatment of sarcopenia and general reduced physical state due to cellular senescence and apoptosis mechanisms.

11. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2 -isopropylphenol according to claims 1 and 2 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according tothe present invention for use in transplantation of progenitor skeletal muscle cells in patients with muscular dystrophies and neurodegenerative diseases for the treatment of reduced regenerative capacity of skeletal muscles.

12. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, hydrate, isotope or salt thereof in any therapeutically effective amount or dosage according to this invention for use in diabetes mellitus for the treatment of diabetic microangiopathy of skeletal muscles, heart, retina and other organs caused by endothelial dysfunction.

13. Pharmaceutical composition according to claim 12 for use in diabetes mellitus and sepsis for the treatment of myopathy caused by endothelial and micro vascular dysfunction.

14. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to this invention for use in the treatment of cardiac dysfunction in clinical ischemiareperfusion conditions, e.g. acute myocardial infarction, coronary vessel angioplasty, donor heart preservation for transplantation, cardiac surgery, in cases where systolic and diastolic functions of the left ventricle of the heart are reduced.

15. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of a patient suffering from cardiac or skeletal muscle dysfunction, in particular after myocardial infarction, sepsis, diabetes, or prolonged exposure to microgravity.

16. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in cardiac diseases, e.g. acute myocardial infarctionand chronic heart failure or skeletal muscle diseases, e.g. musculoskeletal trauma, prolonged immobility and bed confinement, ageing-related sarcopenia where enhancement of the activity of one or more sarcoendoplasmic Reticulum Calcium ATPase pumps (SERCA) pumps is required.

17. Pharmaceutical composition according to claim 16, for use in heart failure for the treatment of reduced left ventricular heart function caused by impaired SERCA2 activity and deregulation of calcium homeostasis.

18. Pharmaceutical composition according to claim 16 for use in musculoskeletal trauma, prolonged immobility, bed confinement, ageing-related sarcopenia and muscular dystrophies for the treatment of skeletal muscle atrophy and weakness, where the SERCA 1 activity is reduced and calcium homeostasis is deregulated.

19. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in microgravity myopathy, musculoskeletal trauma, sepsis myopathy, prolonged immobility, bed confinement and ageing-related sarcopenia for the treatment of deregulation of the intracellular kinase system, in particular p38 Mitogen Activated Protein Kinase and Akt kinase.

20. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate or salt thereof in any therapeutically effective amount or dosage according to the present invention for use in microgravity myopathy, musculoskeletal trauma, sepsis myopathy, myopathy due to prolonged immobility and bed confinement and ageing-related sarcopenia to achieve regulation of the thyroid hormone action at the level of intracellular thyroid hormone receptors in order to promote myogenesis and muscle repair.

21. Pharmaceutical composition comprising the compound 4-(4-((4-Aminobutan- 2yl)oxy)-2,6-dibromophenoxy)-2-isopropylphenol according to claim 1 or a pharmacologically acceptable isomer, racemate, tautomer, isotope, hydrate orsalt thereof in any therapeutically effective amount or dosage according to the present invention for use in the treatment of breast or colon cancer.

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