Pharmaceutical combinations

Abstract

The present invention relates to a combination of suitable dose of suitable COX-2 inhibitor and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 10 mg to 2000 mg. Present invention also provides use of the said combination for the treatment of Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

Description

[0001] PHARMACEUTICAL COMBINATIONS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to a combination of suitable dose of suitable COX-2 inhibitor and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 10 mg to 2000 mg. Present invention also provides use of the said combination for the treatment of Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

[0004] BACKGROUND OF THE INVENTION

[0005] Duchenne muscular dystrophy (DMD) is a genetic muscle-wasting disease and the most common inherited pediatric myopathy, affecting 1 in 3,500-5,000 live male births. It is characterized by progressive muscle weakness and loss of ambulation around age 10 years and is ultimately fatal owing to cardiorespiratory failure around age 30 years. In addition to muscle wasting, commonly observed clinical features of the disease include scoliosis, joint contractures and calf pseudohypertrophy. DMD is caused by mutations that disrupt production of the dystrophin protein and therefore sensitize muscle to contraction-induced damage. Primarily affects the skeletal muscles, which are used for movement, and heart (cardiac) muscle.

[0006] Symptoms: i) Delay in developmental stages like sitting and standing. ii) Difficulty in climbing or raising from a sitting position. iii) Abnormal enlargement of calves muscle. iv) Intellectual disability v) High level of blood creatinine kinase (up to 20 times higher)

[0007] Duchenne muscular dystrophy (DMD) is a rare muscle disorder, but it is one of the most frequent genetic conditions affecting approximately 1 in 3,500 male births worldwide (https: / / rarediseases.org, 2023). It is an X-linked recessive neuromuscular disease caused by loss-of function mutations or deletions in gene encoding dystrophin (Dong X et al. Front. Physiol., 2021). Dystrophin is a plasma membrane protein within the dystrophin-glycoprotein complex (DGC), which forms a bridge between the extracellular matrix and the intracellular cytoskeleton in healthy sarcolemma (Muntoni et al. Lancet Neurol., 2003). In DMD, lack of dystrophin disrupts this bridge and breaks down the membrane integrity of muscle fiber, leading to muscle wasting and degeneration. This childhood disorder is characterized by progressive muscle wasting and loss of ambulation before the teenaged years, followed by respiratory and cardiac complications that ultimately lead to death in the third decade of the patient’s life.

[0008] Till date no curative treatment exists for DMD. The standard care of therapy includes corticosteroids like prednisone and deflazacort. There is still an urgent need to develop appropriate therapies with high efficiency, ease of administration, and wide applicability.

[0009] One possible approach for the treatment of DMD that could benefit all patients while also having systemic effects on affected muscles, is upregulation of the autosomal homolog of dystrophin named utrophin. Reported evidence shows that utrophin can serve as a substitute for dystrophin at the sarcolemma because of their high sequence identity and functional redundancy (Blake et al. Physiol. Rev., 2002).

[0010] Celecoxib is a nonsteroidal anti-inflammatory drug (NSAID) and a specific cyclooxygenase (COX)-2 inhibitor commonly used as an anti-inflammatory treatment in many conditions, such as osteoarthritis and rheumatoid arthritis. Celecoxib showed a marked increase in Utrophin protein level in myotubes and increase the Utrophin level in dystrophic muscle in a mice model (P'eladeau et al, FASEB J., 2018).

[0011] Another approach is to improve the skeletal muscle function by activating the muscle AMPK (activated protein kinase), which reduces skeletal muscle fragility and enhances the myogenesis of slow-twitch, oxidative muscle (Ljubicic et al., 2011). Metformin is a widely used anti-diabetic agent recommended as a first-line oral therapy for type 2 diabetes (T2D) is an indirect agonist of AMPK. In a murine model of Duchenne muscular dystrophy, metformin showed improved sarcolemma integrity and increased muscle strength (Dong X et al. Front. Physiol (2021)).

[0012] Hu et al. disclosed that the combination of Celecoxib and Metformin can be used to improve the antitumor effect by inhibiting the growth of Hepatocellular Carcinoma (Hu et al. J. Cancer (2020)). Animal doses disclosed in this study are 100 mg / kg of Celecoxib and 300 mg / kg of Metformin.

[0013] The inventors of the present invention surprisingly found that combining Celecoxib and Metformin shows significant effects on treating Duchenne Muscular Dystrophy disease.

[0014] SUMMARY OF THE INVENTION

[0015] The present invention relates to the combination of suitable dose of suitable COX- 2 inhibitor and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 10 mg to 2000 mg.. Present invention also provides use of the said combination for the treatment of Duchenne Muscular Dystrophy and Becker muscular dystrophy disease.

[0016] EMBODIMENTS OF THE PRESENT INVENTION

[0017] In an embodiment, the present invention relates to a combination comprising suitable COX-2 inhibitors and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is selected in the range of 50 mg to 2000 mg.

[0018] In an embodiment, the present invention relates to a combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Celecoxib is selected in the range of 10 mg to 500 mg and the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 50 mg to 2000 mg. In another embodiment, the present invention relates to use of a combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts for Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

[0019] In another embodiment, the present invention relates to method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease using a combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts.

[0020] In another embodiment, present invention relates to a pharmaceutical composition comprising a combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts and suitable pharmaceutically acceptable excipients for use in treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease, wherein the dose of Celecoxib is selected in the range of 10 mg to 500 mg and the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 10 mg to 2000 mg.

[0021] DESCRIPTION OF THE INVENTION

[0022] Herein, the term "treating" includes abrogating, substantially inhibiting, slowing or reversing the progression of a disease, substantially ameliorating clinical symptoms of a disease or substantially preventing the appearance of clinical symptoms of a disease. The term ‘pharmaceutically acceptable’ relates to its use for both humans and animals. The term "excipient" or "pharmaceutically acceptable excipient" refers to pharmacologically inactive substances that are added to pharmaceutical preparation in addition to the active pharmaceutical ingredient.

[0023] In an embodiment, the present invention provides a combination of COX-2 inhibitors and Metformin or its suitable pharmaceutically acceptable salts wherein the dose of Metformin, or its suitable pharmaceutically acceptable salts is selected in the range of 50 mg to 2000 mg.

[0024] COX-2 inhibitor used is selected from Celecoxib (Celebrex®), Etoricoxib (Arcoxia®) and parecoxib (Dynastat®). Preferred COX-2 inhibitors is Celecoxib. As described in the above section, the present invention relates to a combination of Celecoxib and Metformin.

[0025] Combination of suitable dose of Celecoxib and suitable dose of Metformin

[0026] In an embodiment, the present invention provides a combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Celecoxib is selected in the range of 10 mg to 500 mg and the dose of Metformin or its suitable pharmaceutically acceptable salts is selected in the range of 50 mg to 2000 mg.

[0027] Suitable pharmaceutically acceptable salt of the Metformin is hydrochloride.

[0028] In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 500 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 400 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 300 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 100 mg. In certain embodiments, combination of the present invention wherein, Celecoxib is in the range of 20 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 30 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 40 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 50 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 60 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 70 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 80 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 90 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 100 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 150 mg to 200 mg.

[0029] In an another embodiment, combination of the present invention wherein Celecoxib used in an amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg,

[0030] 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg,

[0031] 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

[0032] In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 250 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 250 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 700 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 1000 mg to 2000 mg.

[0033] In an another embodiment, combination of the present invention wherein Metformin or it suitable pharmaceutically acceptable salts thereof is in an amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg,

[0034] 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg,

[0035] 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg,

[0036] 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg,

[0037] 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg,

[0038] 1470 mg, 1480 mg, 1490 mg, 1500 mg.

[0039] In a further embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts wherein preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

[0040] Reduction in creatine kinase

[0041] The combination of the present invention is used to reduce the amount of creatine kinase of the subject. For example, in certain embodiments, there is a reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 5% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 10% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 15% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 20% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 25% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 30% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 35% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 40% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 45% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 50% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 55% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 60% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 65% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 70% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 75% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 80% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 85% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 90% w / w reduction in the amount of creatine kinase of the subject. The reduction in the amount of creatine kinase of the subject is with respect to disease control used.

[0042] Reduction in plasma lactate dehydrogenase (LDH)

[0043] The combination of the present invention is used to reduce the amount of plasma LDH of the subject. For example, in certain embodiments, there is a reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 5 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 10 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 15 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 20 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 25 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 30 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 35 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 45 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 50 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 55 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 60 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 65 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 75 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 80 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 85 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 90 % w / w reduction in the amount of plasma LDH of the subject. The reduction in the amount of plasma LDH of the subject is with respect to disease control used.

[0044] In a further embodiment, effective amount of combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject by oral, parenteral, intravenous or intramuscular route of administration.

[0045] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject, wherein subject is animal or human.

[0046] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered as a single composition. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered consecutively. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered simultaneously. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention can be in the form of physical mixture.

[0047] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered once a daily, twice a daily or thrice a daily.

[0048] Use of combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts for the treatment of Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease

[0049] In a certain embodiment, the present invention provides use of combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts for the treatment of Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

[0050] Pharmaceutically acceptable salt of the metformin is hydrochloride.

[0051] In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 500 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 400 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 300 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 100 mg. In certain embodiments, combination of the present invention wherein, Celecoxib is in the range of 20 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 30 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 40 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 50 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 60 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 70 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 80 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 90 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 100 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 150 mg to 200 mg.

[0052] In an another embodiment, combination of the present invention wherein Celecoxib used in an amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg,

[0053] 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg,

[0054] 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

[0055] In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 250 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 250 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 700 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 1000 mg to 2000 mg.

[0056] In an another embodiment, combination of the present invention wherein Metformin or it suitable pharmaceutically acceptable salts thereof is in an amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg,

[0057] 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg,

[0058] 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg,

[0059] 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg,

[0060] 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg,

[0061] 1470 mg, 1480 mg, 1490 mg, 1500 mg.

[0062] In a further embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts wherein preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

[0063] Reduction in creatine kinase

[0064] The combination of the present invention is used to reduce the amount of creatine kinase of the subject. For example, in certain embodiments, there is a reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 5% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 10% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 15% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 20% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 25% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 30% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 35% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 40% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 45% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 50% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 55% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 60% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 65% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 70% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 75% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 80% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 85% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 90% w / w reduction in the amount of creatine kinase of the subject. The reduction in the amount of creatine kinase of the subject is with respect to disease control used.

[0065] Reduction in plasma lactate dehydrogenase (LDH)

[0066] The combination of the present invention is used to reduce the amount of plasma LDH of the subject. For example, in certain embodiments, there is a reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 5 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 10 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 15 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 20 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 25 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 30 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 35 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 45 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 50 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 55 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 60 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 65 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 75 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 80 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 85 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 90 % w / w reduction in the amount of plasma LDH of the subject. The reduction in the amount of plasma LDH of the subject is with respect to disease control used.

[0067] In a further embodiment, effective amount of combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject by oral, parenteral, intravenous or intramuscular route of administration.

[0068] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject, wherein subject is animal or human.

[0069] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered as a single composition. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered consecutively. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered simultaneously. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention can be in the form of physical mixture.

[0070] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered once a daily, twice a daily or thrice a daily.

[0071] Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease using a combination comprising Celecoxib and Metformin

[0072] In a certain embodiment, the present invention provides method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease using a combination comprising Celecoxib and Metformin.

[0073] Pharmaceutically acceptable salt of the metformin is hydrochloride.

[0074] In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 500 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 400 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 300 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 10 mg to 100 mg. In certain embodiments, combination of the present invention wherein, Celecoxib is in the range of 20 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 30 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 40 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 50 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 60 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 70 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 80 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 90 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 100 mg to 200 mg. In certain embodiments, combination of the present invention wherein Celecoxib is in the range of 150 mg to 200 mg.

[0075] In an another embodiment, combination of the present invention wherein Celecoxib used in an amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg,

[0076] 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg,

[0077] 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

[0078] In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 250 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 250 mg to 2000 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 700 mg to 1500 mg. In certain embodiments, combination of the present invention wherein metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 1000 mg to 2000 mg.

[0079] In an another embodiment, combination of the present invention wherein Metformin or it suitable pharmaceutically acceptable salts thereof is in an amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg,

[0080] 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg,

[0081] 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg,

[0082] 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg,

[0083] 1470 mg, 1480 mg, 1490 mg, 1500 mg.

[0084] In a further embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts wherein preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

[0085] Reduction in creatine kinase

[0086] The combination of the present invention is used to reduce the amount of creatine kinase of the subject. For example, in certain embodiments, there is a reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 5% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 10% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 15% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 20% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 25% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 30% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 35% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 40% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 45% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 50% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 55% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 60% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 65% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 70% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 75% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 80% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 85% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 90% w / w reduction in the amount of creatine kinase of the subject. The reduction in the amount of creatine kinase of the subject is with respect to disease control used.

[0087] Reduction in plasma lactate dehydrogenase (LDH)

[0088] The combination of the present invention is used to reduce the amount of plasma LDH of the subject. For example, in certain embodiments, there is a reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 5 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 10 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 15 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 20 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 25 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 30 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 35 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 45 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 50 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 55 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 60 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 65 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 75 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 80 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 85 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 90 % w / w reduction in the amount of plasma LDH of the subject. The reduction in the amount of plasma LDH of the subject is with respect to disease control used.

[0089] In a further embodiment, effective amount of combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject by oral, parenteral, intravenous or intramuscular route of administration.

[0090] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered to a subject, wherein subject is animal or human. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered as a single composition. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered consecutively. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered simultaneously. In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention can be in the form of physical mixture.

[0091] In a certain embodiment, combination of Celecoxib and metformin or its suitable pharmaceutically acceptable salts of the present invention is administered once a daily, twice a daily or thrice a daily.

[0092] Pharmacological data:

[0093] Efficacy of Celecoxib in combination with Metformin in Duchenne muscular dystrophy (DMD) in mdx mice.

[0094] Duchenne muscular dystrophy (DMD) is a rare muscle disorder, but it is one of the most frequent genetic conditions affecting approximately 1 in 3,500 male births worldwide (https: / / rarediseases.org / rare-diseases / duchenne-muscular-dystrophy / ). It is an X-linked recessive neuromuscular disease caused by loss-of function mutations or deletions in gene encoding dystrophin (Dong, et al. Front. Physiol., 2021). Dystrophin is a plasma membrane protein within the dystrophin- glycoprotein complex (DGC), which forms a bridge between the extracellular matrix and the intracellular cytoskeleton in healthy sarcolemma (Muntoni, et al., Lancet Neurol., 2003). In DMD, lack of dystrophin disrupts this bridge and breaks down the membrane integrity of muscle fiber, leading to muscle wasting and degeneration. This childhood disorder is characterized by progressive muscle wasting and loss of ambulation before the teenaged years, followed by respiratory and cardiac complications that ultimately lead to death in the third decade of the patient’s life. Assessments of disease progression and response to therapies in Duchenne muscular dystrophy (DMD) patients remain challenging. Current standardized outcome measures used in DMD clinical trials include a walk test i.e., distance covered by patient in a specific time as well as quantitative muscle strength tests. Molecular biomarkers measured in blood could be proven valuable to assess disease progression and response to therapies in DMD patients. These included creatine kinases (CK), a muscle specific protein that reflects sarcolemma damage. CK was reported a long ago to be elevated in serum of DMD patients relative to controls and it is currently used to screen for DMD in newborns (Hathout et al., Clin Proteom. 2016). Alongside CK the lactate dehydrogenase (LDH) is also significantly increased in DMD patients (Zhu, et al., Dis Markers., 2015).

[0095] Till date no curative treatment exists for DMD. The standard care of therapy includes corticosteroids like prednisone and deflazacort. Some gene therapies has been approved by FDA like Exondys 51, Vyondys 53 and Viltepso (https: / / rarediseases.org / rare-diseases / duchenne-muscular-dystrophy / ). There is still an urgent need to develop appropriate therapies with high efficiency, ease of administration, and wide applicability.

[0096] One possible approach for the treatment of DMD that could benefit all patients while also having systemic effects on affected muscles, is upregulation of the autosomal homolog of dystrophin named utrophin. Reported evidence show that utrophin can serve as a substitute for dystrophin at the sarcolemma because of their high sequence identity and functional redundancy (Blake, et al., Physiol. Rev., 2002).

[0097] Celecoxib, a COX-2 inhibitor, has shown promising results beyond its traditional use as an anti-inflammatory drug. Research indicates that it can significantly increase Utrophin protein levels in myotubes and dystrophic muscles in mice models. This is particularly relevant for conditions like Duchenne Muscular Dystrophy (DMD), where Utrophin upregulation could potentially compensate for the lack of dystrophin (P'eladeau, et al., FASEB J., 2018).

[0098] Another approach is to improve the skeletal muscle function by activating the muscle AMPK, which reduces skeletal muscle fragility and enhances the myogenesis of slow-twitch, oxidative muscle (Ljubicic, et al., Hum. Mol. Genet., Metformin is a widely used anti-diabetic agent recommended as a first-line oral therapy for type 2 diabetes (T2D) is an indirect agonist of AMPK. The findings from murine models suggest that metformin helps improve sarcolemma (the muscle cell membrane) integrity, which is critical for maintaining muscle function, and enhances muscle strength (Dong, et al. Front. Physiol., 2021).

[0099] Combining both approach i.e. increasing the Utrophin level and improving the sarcolemma integrity by using a fixed dose combination of Celecoxib and Metformin may have some therapeutic advantage on DMD over existing therapies. To study the combination effect of Celecoxib and Metformin we used the dystrophin deficient mdx mouse, which is a well-known mouse model for DMD with a characteristic of muscle weakness (Bulfield, et al., Proc. Natl. Acad. Sci. U.S.A., 1984).

[0100] Methods

[0101] In vivo experiments were performed using mdx mice with spontaneous mutation in the dystrophin gene which is a widely used mice model for studying Duchenne muscular dystrophy. Mdx mice at the age of 10-1 Iweek age were used in this study. One group of age matched C57 (wild-type) mice were taken and serve as the normal control. On the day of study initiation (day-0) body weight of all the animals were recorded, the animals were subjected to a treadmill exercise and forelimb grip strength was measure using a grip-strength meter. Based on the forelimb gripstrength and body weight, the animals were randomized in to following treatment groups.

[0102] The test compounds were formulated in using PEG400, Tween80 and 0.5% sodium salt of carboxy methyl cellulose in water in a ratio of 5:5:90 as vehicle. Formulations were administered once daily orally at 10 ml / kg dose for 4 weeks. During the study period body weight was measured twice weekly and forelimb gripstrength was measured once weekly after treadmill exercise till the end of study. The forelimb grip-strength was represented as the ratio of forelimb grip-strength to body weight. At the end of the treatment period i.e. on completion of week 4, animals were subjected to an exhaustion treadmill test and the time to exhaust and the distance travelled were recorded. After the treadmill test forelimb grip strength was measured and blood collection was performed under isoflurane anesthesia for muscle damage biomarkers like plasma creatine kinase (CK) and lactate dehydrogenase (LDH) and animals were sacrificed by CO2 asphyxiation. Skeletal muscles like gastrocnemius muscle and tibialis anterior muscle were collected and weighed. One part of each collected skeletal muscle were stored for muscle utrophin level and gene expression while another part was stored for tissue histology.

[0103] Results:

[0104] At the end of study period, the disease control animals showed a significantly reduced forelimb grip-strength and significantly high plasma creatin kinase (CK) as well as plasma lactate dehydrogenase (LDH) which are the biomarkers for muscle damage. The combination of significantly high plasma CK and LDH levels, along with reduced forelimb grip strength, strongly indicates the presence of established Duchenne muscular dystrophy (DMD) in the disease control animals.

[0105] Table 1: Treatment groups and dose levels

[0106] Effect on forelimb grip-strength:

[0107] The forelimb grip-strength is represented as the grip-strength to body weight ratio. Treatment with metformin alone showed a non-significant 12.7% normalization and celecoxib has alone showed -1.2% normalization in forelimb grip-strength as compared to disease control. When both metformin and celecoxib were administered as a combined dose form, the forelimb grip-strength was normalized to 53.9% which is significant as compared to disease control (Table 2).

[0108] Table 2: Effect on forelimb grip-strength to body weight ratio

[0109] * Significantly different from disease control at p<0.05 using unpaired t-test.

[0110] Effect on distance travelled during exhaustion treadmill test:

[0111] One of the major parameters in considering endurance is the “distance travelled” during exhaustion treadmill test. Treatment with metformin alone showed a 25.9% reduction in the “distance travelled’ whereas celecoxib alone showed a 15.8% increase in the “distance travelled’ during exhaustion treadmill test. Combination of metformin and celecoxib treatment showed a significant 77.1% increase in the “distance travelled’ as compared to disease control during the exhaustion treadmill test. (Table 3)

[0112] Table 3: Effect on distance travelled during exhaustion treadmill test

[0113] ** Significantly different from disease control at p<0.01 using unpaired t-test Effect on ‘time to exhaust’ during exhaustion treadmill test:

[0114] “Time to exhaust” is the time when the animal stop moving during the exercise which is an indicator in accessing endurance during treadmill exercise. During exhaustion treadmill test, metformin alone treated mice showed 21.3% reduction in “time to exhaust” whereas celecoxib alone treated mice showed 15.2% increase in the “time to exhaust” as compared to disease control group. When metformin and celecoxib were administered in combination, a significant 70.1% increase in “time to exhaust” (Table 4) as compared to disease control.

[0115] Table 4: Effect on “time to exhaust” during exhaustion treadmill test

[0116] * Significantly different from disease control at p<0.05 using one way ANOVA followed by Dunnett multiple comparison method

[0117] Effect on plasma biomarker creatine kinase (CK):

[0118] The disease control animals showed a 916-fold high level of plasma CK level as compared to normal control animals. Treatment with metformin alone showed 13.4% rise and celecoxib alone showed 31.4% reduction in the plasma CK level as compared to disease control. Metformin and celecoxib in combined dosage form resulted in a significant 58.2% reduction in plasma CK level (Table 5).

[0119] Table 5: Effect on plasma biomarker creatine kinase

[0120] ** Significantly different from disease control at p<0.01 using one-way ANOVA followed by Dunnetts multiple comparison method

[0121] Effect on plasma biomarker lactate dehydrogenase (LDH):

[0122] The disease control animal showed a 65-fold high level of plasma LDH as compared to normal control group. Metformin and celecoxib alone treated group showed 9.1% and 8.7% reduction in the plasma LDH level respectively as compared to disease control group. When metformin and celecoxib were administered as a combine dosage form, it resulted in a significant 53.6 % reduction in LDH level as compared to disease control group (Table 6).

[0123] Table 6: Effect on plasma biomarker lactate dehydrogenase.

[0124] ** Significantly different from disease control at p<0.01 using one way ANOVA followed by Dunnett multiple comparison method

[0125] Conclusion:

[0126] In conclusion, combination of metformin and celecoxib improved the DMD condition in mdx mice by improving muscle grip strength and endurance, as well as reduced muscle damage biomarkers like plasma CK and LDH. While metformin or celecoxib alone did not show any significant improvement in the DMD condition, the combined dosage form showed a synergistic improvement in DMD as compared to either of the treatment alone in the mdx mice model of Duchenne muscular dystrophy.

[0127] A pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts and pharmaceutically acceptable excipients for use in treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease

[0128] In another embodiment, present invention provides pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts and pharmaceutically acceptable excipients for use in treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease. Pharmaceutically acceptable salt of the metformin is hydrochloride.

[0129] Pharmaceutically acceptable excipients can be selected from diluents, binders, emulsifiers, disintegrating agents, lubricants, glidants, buffering agents, tonicity agents, preservatives, chelating agents, solubilizing agents, plasticizer, opacifier, film forming agents and the like. Examples of excipient classes frequently used are listed below.

[0130] Non-limiting examples of diluents include but not limited to starch and its processed and co-processed derivatives, saccharides, di saccharides, sucrose, lactose, polysaccharides, cellulose, cellulose ethers, cellulose acetate, hydroxypropyl cellulose, sugar alcohols, xylitol, sorbitol, maltitol, lactitol, microcrystalline cellulose, magnesium or calcium or sodium carbonate, lactose, lactose monohydrate, di-calcium phosphate, compressible sugars, di-basic calcium phosphate dihydrate, mannitol lactose anhydrous, magnesium oxide, maltodextrin, maltose, pullulan, sodium alginate, sodium bicarbonate, calcium silicate, calcium sulphate, cell and tribasic calcium phosphate or suitable combinations thereof.

[0131] Non-limiting examples of emulsifiers include but not limited to sodium stearoyl lactylate, mono- and di-glycerols, Polysorbate 80 (tween 80), ammonium phosphatide, locust bean gum, and xanthan gum or suitable combinations thereof.

[0132] Non-limiting examples of binders include but not limited to chitosan, hydrogenated castor oil, sodium alginate, carbomers, cellulose acetate phthalate, povidone, sugar, hydroxypropylmethyl-cellulose, hydroxypropylcellulose, carboxymethyl cellulose, starch, alginic acid, pre-gelatinized starch, acacia, tragakanth, ethylcellulose, acrylic and methacrylic acid co-polymers or suitable combinations thereof.

[0133] Non-limiting examples of disintegrants include but not limited to maize starch, sodium starch glycolate, croscarmellose sodium, crospovidone, microcrystalline cellulose, modified com starch, sodium carboxymethyl starch, povidone, pregelatinized starch, agar, carboxymethyl cellulose calcium or sodium, colloidal silicon dioxide, chitosan, docusate sodium, hydroxyl propyl cellulose, magnesium aluminium silicate, maltose, methyl cellulose, polacrilin potassium, and alginic acid or suitable combinations thereof.

[0134] Non -limiting examples of lubricants include but not limited to magnesium stearate, stearic acid, silica, fats, zinc or sucrose or sodium or calcium stearate, castor oil, hydrogenated castor oil, Polyethylene glycol and its derivatives, sodium stearyl fumarate, talc, or fatty acids including lauric acid, oleic acid, glyceryl behenate, glyceryl monostearate and Cl -CIO fatty acid or suitable combinations thereof.

[0135] Non-limiting examples of glidants include but not limited to colloidal silicon dioxide, talc, fumed silica, starch, starch derivatives, and bentonite or suitable combinations thereof.

[0136] Non-limiting examples of plasticizer include but not limited to polyols like polyethylene glycols, propylene glycol, glycerol (glycerin), organic esters like phthalate esters (diethyl, dibutyl), dibutyl sebacete, citrate esters (triethyl, acetyl triethyl, acetyl tributyl), triacetin, Oils / glycerides like castor oil; acetylated monoglycerides, fractionated coconut oil or suitable combinations thereof.

[0137] Non-limiting examples of opacifier include but not limited to titanium dioxide, talc, sunset yellow, tartrazine, erythrosine, iron oxide yellow, red and black, carmine, anthocyanins, allura Red AC, allura Red AC aluminum lake, indigotine, indigotine aluminum lake or suitable combinations thereof.

[0138] Non-limiting examples of film forming agents include but not limited to hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, hydroxypropyl cellulose, povidone, polydextrose, lactose, maltodextrin, acrylic polymer or suitable combinations thereof.

[0139] One or more solvents or solubilizing agents used in the formulation are selected from water, acetone, chloroform, dichloromethane, ethyl alcohol, ethyl acetate, methyl alcohol, isopropyl alcohol, N, N-dimethyl formamide and combinations thereof and other such materials known to those of ordinary skill in the art. Non-limiting examples of buffering agents include but not limited to sodium phosphate, acetic acid, citric acid, histamine, glycine, tartaric acid, tromethamine and other suitable pharmaceutically acceptable buffers and combinations thereof known to those of ordinary skill in the art.

[0140] Non-limiting examples of tonicity agents include but not limited to mannitol, lactose, dextrose, sodium chloride, sorbitol and suitable combinations thereof known to those of ordinary skill in the art.

[0141] Non-limiting examples of preservatives include but not limited to phenol and benzyl alcohol derivatives such as benzyl alcohol, chlorobutanol, m-cresol, methylparaben, phenol, phenoxyethanol, propylparaben, thimerosal and suitable combinations thereof known to those of ordinary skill in the art.

[0142] Non-limiting examples of chelating agents include but not limited to disodium EDTA, Calcium disodium EDTA, tetrasodium EDTA and suitable combinations thereof known to those of ordinary skill in the art.

[0143] In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 10 mg to 500 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 10 mg to 400 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 10 mg to 300 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 10 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 10 mg to 100 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 20 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 30 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 40 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 50 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 60 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 70 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 80 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 90 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 100 mg to 200 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib is in the range of 150 mg to 200 mg.

[0144] In an another embodiment, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Celecoxib used in an amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

[0145] In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 2000 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1500 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 1000 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 500 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 50 mg to 250 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 250 mg to 2000 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 700 mg to 1500 mg. In certain embodiments, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or its suitable pharmaceutically acceptable salts thereof is in the range of 1000 mg to 2000 mg.

[0146] In an another embodiment, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein Metformin or it suitable pharmaceutically acceptable salts thereof is in an amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg,

[0147] 1220 mg, 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg,

[0148] 1300 mg, 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg,

[0149] 1380 mg, 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg,

[0150] 1460 mg, 1470 mg, 1480 mg, 1490 mg, 1500 mg.

[0151] In a further embodiment, a pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts wherein preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

[0152] Reduction in creatine kinase

[0153] The pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is used to reduce the amount of creatine kinase of the subject. For example, in certain embodiments, there is a reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 5% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 10% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 15% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 20% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 25% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 30% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 35% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 40% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 45% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 50% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 55% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 60% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 65% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 70% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 75% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 80% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least an 85% w / w reduction in the amount of creatine kinase of the subject. In certain embodiments, there is at least a 90% w / w reduction in the amount of creatine kinase of the subject.

[0154] Reduction in plasma lactate dehydrogenase (LDH)

[0155] The pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is used to reduce the amount of plasma LDH of the subject. For example, in certain embodiments, there is a reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 5 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 10 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 15 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 20 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 25 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 30 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 35 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 45 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 50 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 55 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 60 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 65 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 40 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 75 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 80 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least an 85 % w / w reduction in the amount of plasma LDH of the subject. In certain embodiments, there is at least a 90 % w / w reduction in the amount of plasma LDH of the subject.

[0156] In a further embodiment, effective amount of pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered to a subject by oral, parenteral, intravenous or intramuscular route of administration.

[0157] In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered to a subject, wherein subject is animal or human.

[0158] In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered as a single composition. In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered consecutively. In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered simultaneously. In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts can be in the form of physical mixture.

[0159] In a certain embodiment, pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts is administered once a daily, twice a daily or thrice a daily.

[0160] The invention is exemplified by the non-limiting examples, which are illustrative representing the preferred modes of carrying out the invention. The invention’s scope is not limited to these specific embodiments only but should be read in conjunction with what is disclosed anywhere else in the specification together with that information and knowledge which are within the general understanding of a person skilled in the art.

[0161] The compositions are prepared and formulated according to conventional methods, such as those disclosed in standard reference texts and are well within the scope of a skilled person.

[0162] References:

[0163] 1. Dong X, Hui T, Chen J, Yu Z, Ren D, Zou S, Wang S, Fei E, Jiao H and Lai X (2021) Metformin Increases Sarcolemma Integrity and Ameliorates Neuromuscular Deficits in a Murine Model of Duchenne Muscular Dystrophy. Front. Physiol. 12:642908.

[0164] 2. Muntoni, F., Torelli, S., and Ferlini, A. (2003). Dystrophin and mutations: one gene, several proteins, multiple phenotypes. Lancet Neurol. 2, 731— 740.

[0165] 3. Hathout, Y., Seol, H., Han, M.H.J. et al. Clinical utility of serum biomarkers in Duchenne muscular dystrophy. Clin Proteom 13, 9 (2016). Zhu Y, Zhang H, Sun Y, Li Y, Deng L, Wen X, Wang H, Zhang C (2015) Serum enzyme profiles differentiate five types of muscular dystrophy. Dis Markers 2015:543282 Blake, D. J.,Weir,A.,Newey, S.E., andDavies, K. E. (2002)Function and genetics of dystrophin and dystrophin-related proteins in muscle. Physiol. Rev. 82, 291-329. P'eladeau, C., Adam,N. J., Jasmin, B. J. Celecoxib treatment improves muscle function in mdx mice and increases utrophin A expression. FASEB J. 32, 000-000 (2018). Ljubicic, V., Miura, P., Burt, M., Boudreault, L., Khogali, S., Lunde, J. A., Renaud, J. M., and Jasmin, B. J. (2011) Chronic AMPK activation evokes the slow, oxidative myogenic program and triggers beneficial adaptations in mdx mouse skeletal muscle. Hum. Mol. Genet. 20, 3478-3493. Bulfield, G. S. W ., Wight, P. A., and Moore, K. J. (1984). X chromosome- linked muscular dystrophy (mdx) in the mouse. Proc. Natl. Acad. Sci. U.S.A. 81, 1189-1192. doi: 10.1073 / pnas.81.4.1189.

Claims

We claim:

1. A combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts, wherein the dose of Celecoxib is selected in the range of 10 mg to 500 mg and the dose of Metformin or its suitable pharmaceutically acceptable salt is selected in the range of 50 mg to 2000 mg.

2. A combination as claimed in claim 1 wherein suitable pharmaceutically acceptable salt of the Metformin is hydrochloride.

3. A combination as claimed in claim 1 wherein the dose of Celecoxib is in the range of 10 mg to 500 mg, 10 mg to 400 mg, 10 mg to 300 mg, 10 mg to 200 mg, 10 mg to 100 mg, 20 mg to 200 mg, 30 mg to 200 mg, 40 mg to 200 mg, 50 mg to 200 mg, 60 mg to 200 mg, 70 mg to 200 mg, 80 mg to 200 mg, 90 mg to 200 mg, 100 mg to 200 mg, 150 mg to 200 mg.

4. A combination as claimed in claim 1 wherein the dose of Celecoxib is in amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

5. A combination as claimed in claim 1 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in the range of 50 mg to 2000 mg, 50 mg to 1500 mg, 50 mg to 1000 mg, 50 mg to 500 mg, 50 mg to 250 mg, 250 mg to 2000 mg, 700 mg to 1500 mg, 1000 mg to 2000 mg.

6. A combination as claimed in claim 1 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in amount of 700 mg, 710 mg,720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg,890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg, 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg, 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg, 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg, 1470 mg, 1480 mg, 1490 mg, 1500 mg.

7. A combination as claimed in claim 1 wherein the preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

8. A combination as claimed in claim 1 wherein creatine kinase of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

9. A combination as claimed in claim 1 wherein plasma LDH of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

10. Use of combination comprising Celecoxib and Metformin or its suitable pharmaceutically acceptable salts for the treatment of Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

11. Use of combination as claimed in claim 10 wherein suitable pharmaceutically acceptable salt of the Metformin is hydrochloride.

12. Use of combination as claimed in claim 10 wherein the dose of Celecoxib is in the range of 10 mg to 500 mg, 10 mg to 400 mg, 10 mg to 300 mg, 10 mg to 200 mg, 10 mg to 100 mg, 20 mg to 200 mg, 30 mg to 200 mg, 40 mg to 200 mg, 50 mg to 200 mg, 60 mg to 200 mg, 70 mg to 200 mg, 80 mg to 200 mg, 90 mg to 200 mg, 100 mg to 200 mg, 150 mg to 200 mg.

13. Use of combination as claimed in claim 10 wherein the dose of Celecoxib is in amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg,27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg,152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

14. Use of combination as claimed in claim 10 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in the range of 50 mg to 2000 mg, 50 mg to 1500 mg, 50 mg to 1000 mg, 50 mg to 500 mg, 50 mg to 250 mg, 250 mg to 2000 mg, 700 mg to 1500 mg, 1000 mg to 2000 mg.

15. Use of combination as claimed in claim 10 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg, 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg, 1310 mg, 1320 mg, 1330 mg, 1340 mg,1350 mg, 1360 mg, 1370 mg, 1380 mg, 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg, 1470 mg, 1480 mg, 1490 mg, 1500 mg.

16. Use of combination as claimed in claim 10 wherein the preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

17. Use of combination as claimed in claim 10 wherein creatine kinase of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

18. Use of combination as claimed in claim 10 wherein plasma LDH of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

19. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease using a combination comprising Celecoxib and Metformin.

20. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein suitable pharmaceutically acceptable salt of the Metformin is hydrochloride.

21. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein the dose of Celecoxib is in the range of 10 mg to 500 mg, 10 mg to 400 mg, 10 mg to 300 mg, 10 mg to 200 mg, 10 mg to 100 mg, 20 mg to 200 mg, 30 mg to 200 mg, 40 mg to 200 mg, 50 mg to 200 mg, 60 mg to 200 mg, 70 mg to 200 mg, 80 mg to 200 mg, 90 mg to 200 mg, 100 mg to 200 mg, 150 mg to 200 mg.

22. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein the dose of Celecoxib is in amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

23. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in the range of 50 mg to 2000 mg, 50 mg to 1500 mg, 50 mg to 1000 mg, 50 mg to 500 mg, 50 mg to 250 mg, 250 mg to 2000 mg, 700 mg to 1500 mg, 1000 mg to 2000 mg.

24. Method of treating Duchenne Muscular Dystrophy and Becker MuscularDystrophy disease as claimed in claim 19 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg,800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg,970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg,1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg, 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg, 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg, 1390 mg, 1400 mg, 1410 mg, 1420mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg, 1470 mg, 1480 mg, 1490 mg, 1500 mg.

25. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein the preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

26. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein creatine kinase of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

27. Method of treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease as claimed in claim 19 wherein plasma LDH of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

28. A pharmaceutical composition comprising combination of Celecoxib and Metformin or its suitable pharmaceutically acceptable salts and pharmaceutically acceptable excipients for use in treating Duchenne Muscular Dystrophy and Becker Muscular Dystrophy disease.

29. A pharmaceutical composition as claimed in claim 28 wherein Metformin or its suitable pharmaceutically acceptable salt is selected from hydrochloride.

30. A pharmaceutical composition as claimed in claim 28 wherein pharmaceutically acceptable excipients are selected from diluents, binders, emulsifiers, disintegrating agents, lubricants, glidants, buffering agents,tonicity agents, preservatives, chelating agents, solubilizing agents, plasticizer, opacifier and film forming agents.

31. A pharmaceutical composition as claimed in claim 30 wherein diluents are selected from starch and its processed and co-processed derivatives, saccharides, di saccharides, sucrose, lactose, polysaccharides, cellulose, cellulose ethers, cellulose acetate, hydroxypropyl cellulose, sugar alcohols, xylitol, sorbitol, maltitol, lactitol, microcrystalline cellulose, magnesium or calcium or sodium carbonate, lactose, lactose monohydrate, di-calcium phosphate, compressible sugars, di-basic calcium phosphate dihydrate, mannitol lactose anhydrous, magnesium oxide, maltodextrin, maltose, pullulan, sodium alginate, sodium bicarbonate, calcium silicate, calcium sulphate, cell and tribasic calcium phosphate or suitable combinations thereof; binders are selected from chitosan, hydrogenated castor oil, sodium alginate, carbomers, cellulose acetate phthalate, povidone, sugar, hydroxypropylmethyl-cellulose, hydroxypropylcellulose, carboxymethyl cellulose, starch, alginic acid, pre-gelatinized starch, acacia, tragakanth, ethylcellulose, acrylic and methacrylic acid co-polymers or suitable combinations thereof; emulsifiers are selected from sodium stearoyl lactylate, mono- and diglycerols, Polysorbate 80 (tween 80), ammonium phosphatide, locust bean gum, and xanthan gum or suitable combinations thereof; disintegrants are selected from maize starch, sodium starch glycolate, croscarmellose sodium, crospovidone, microcrystalline cellulose, modified com starch, sodium carboxymethyl starch, povidone, pregelatinized starch, agar, carboxymethyl cellulose calcium or sodium, colloidal silicon dioxide, chitosan, docusate sodium, hydroxyl propyl cellulose, magnesium aluminium silicate, maltose, methyl cellulose, polacrilin potassium, and alginic acid or suitable combinations thereof;lubricants are selected from magnesium stearate, stearic acid, silica, fats, zinc or sucrose or sodium or calcium stearate, castor oil, hydrogenated castor oil, Polyethylene glycol and its derivatives, sodium stearyl fumarate, talc, or fatty acids including lauric acid, oleic acid, glyceryl behenate, glyceryl monostearate and Cl -CIO fatty acid or suitable combinations thereof; glidants are selected from colloidal silicon dioxide, talc, fumed silica, starch, starch derivatives, and bentonite or suitable combinations thereof; plasticizer are selected from polyols like polyethylene glycols, propylene glycol, glycerol (glycerin), organic esters like phthalate esters (diethyl, dibutyl), dibutyl sebacete, citrate esters (triethyl, acetyl triethyl, acetyl tributyl), triacetin, Oils / glycerides like castor oil; acetylated monoglycerides, fractionated coconut oil or suitable combinations thereof; opacifier are selected from titanium dioxide, talc, sunset yellow, tartrazine, erythrosine, iron oxide yellow, red and black, carmine, anthocyanins, allura Red AC, allura Red AC aluminum lake, indigotine, indigotine aluminum lake or suitable combinations thereof; film forming agents are selected from hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, hydroxypropyl cellulose, povidone, polydextrose, lactose, maltodextrin, acrylic polymer or suitable combinations thereof; solubilizing agents are selected from water, acetone, chloroform, dichloromethane, ethyl alcohol, ethyl acetate, methyl alcohol, isopropyl alcohol, N, N-dimethyl formamide and combinations thereof; buffering agents are selected from sodium phosphate, acetic acid, citric acid, histamine, glycine, tartaric acid, tromethamine and other suitable pharmaceutically acceptable buffers and combinations thereof; tonicity agents are selected from mannitol, lactose, dextrose, sodium chloride, sorbitol and suitable combinations thereof;preservatives are selected from phenol and benzyl alcohol derivatives such as benzyl alcohol, chlorobutanol, m-cresol, methylparaben, phenol, phenoxyethanol, propylparaben, thimerosal and suitable combinations thereof; chelating agents are selected from disodium EDTA, Calcium disodium EDTA, tetrasodium EDTA and suitable combinations thereof.

32. A pharmaceutical composition as claimed in claim 28 wherein the dose of Celecoxib is in the range of 10 mg to 500 mg, 10 mg to 400 mg, 10 mg to 300 mg, 10 mg to 200 mg, 10 mg to 100 mg, 20 mg to 200 mg, 30 mg to 200 mg, 40 mg to 200 mg, 50 mg to 200 mg, 60 mg to 200 mg, 70 mg to 200 mg, 80 mg to 200 mg, 90 mg to 200 mg, 100 mg to 200 mg, 150 mg to 200 mg.

33. A pharmaceutical composition as claimed in claim 28 wherein the dose of Celecoxib is in amount of 10 mg, 12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, 25 mg, 27.5 mg, 30 mg, 32.5 mg, 35 mg, 37.5 mg, 40 mg, 42.5 mg, 45 mg, 47.5 mg, 50 mg, 52.5 mg, 55 mg, 57.5 mg, 60 mg, 62.5 mg, 65 mg, 67.5 mg, 70 mg, 72.5 mg, 75 mg, 77.5 mg, 80 mg, 82.5 mg, 85 mg, 87.5 mg, 90 mg, 92.5 mg, 95 mg, 97.5 mg, 100 mg, 102.5 mg, 105 mg, 107.5 mg, 110 mg, 112.5 mg, 115 mg, 117.5 mg, 120 mg, 122.5 mg, 125 mg, 127.5 mg, 130 mg, 132.5 mg, 135 mg, 137.5 mg, 140 mg, 142.5 mg, 145 mg, 147.5 mg, 150 mg, 152.5 mg, 155 mg, 157.5 mg, 160 mg, 162.5 mg, 165 mg, 167.5 mg, 170 mg, 172.5 mg, 175 mg, 177.5 mg, 180 mg, 182.5 mg, 185 mg, 187.5 mg, 190 mg, 192.5 mg, 195 mg, 197.5 mg, 200 mg.

34. A pharmaceutical composition as claimed in claim 28 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in the range of 50 mg to 2000 mg, 50 mg to 1500 mg, 50 mg to 1000 mg, 50 mg to 500 mg, 50 mg to 250 mg, 250 mg to 2000 mg, 700 mg to 1500 mg, 1000 mg to 2000 mg.

35. A pharmaceutical composition as claimed in claim 28 wherein the dose of Metformin or its suitable pharmaceutically acceptable salts is in amount of 700 mg, 710 mg, 720 mg, 730 mg, 740 mg, 750 mg, 760 mg, 770 mg, 780 mg, 790 mg, 800 mg, 810 mg, 820 mg, 830 mg, 840 mg, 850 mg, 860 mg, 870 mg, 880 mg, 890 mg, 900 mg, 910 mg, 920 mg, 930 mg, 940 mg, 950 mg, 960 mg, 970 mg, 980 mg, 990 mg, 1000 mg, 1010 mg, 1020 mg, 1030 mg, 1040 mg, 1050 mg, 1060 mg, 1070 mg, 1080 mg, 1090 mg, 1100 mg, 1110 mg, 1120 mg, 1130 mg, 1140 mg, 1150 mg, 1160 mg, 1170 mg, 1180 mg, 1190 mg, 1200 mg, 1210 mg, 1220 mg, 1230 mg, 1240 mg, 1250 mg, 1260 mg, 1270 mg, 1280 mg, 1290 mg, 1300 mg, 1310 mg, 1320 mg, 1330 mg, 1340 mg, 1350 mg, 1360 mg, 1370 mg, 1380 mg, 1390 mg, 1400 mg, 1410 mg, 1420 mg, 1430 mg, 1440 mg, 1450 mg, 1460 mg, 1470 mg, 1480 mg, 1490 mg, 1500 mg.

36. A pharmaceutical composition as claimed in claim 28 wherein the preferred dose of Celecoxib is in the range of 75 mg to 150 mg and Metformin or its suitable pharmaceutically acceptable salts is in the range of 800 mg to 1500 mg.

37. A pharmaceutical composition as claimed in claim 28 wherein creatine kinase of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

38. A pharmaceutical composition as claimed in claim 28 wherein plasma LDH of the subject is reduced in the amount of 5 %w / w, 10 %w / w, 15% w / w, 20% w / w, 25% w / w, 30% w / w, 35% w / w, 40% w / w, 45% w / w, 50% w / w, 55% w / w, 60% w / w, 65% w / w, 70% w / w, 75% w / w, 80% w / w, 85% w / w and 90% w / w.

39. A pharmaceutical composition as claimed in claim 28 wherein pharmaceutical composition is administered to a Subject by oral, parenteral, intravenous or intramuscular route of administration.

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