Treatment for Muscular Dystrophy

Abstract

A new dosing regimen is described for treating muscular dystrophy in patients suffering from Duchenne muscular dystrophy (DMD) using antisense oligonucleotide conjugates that cause exon skipping in the human dystrophin gene.Also described is a method for treating patients using antisense oligomeric CPP conjugates and magnesium supplements.Also provided is a method for reducing toxicity associated with treatment involving antisense oligonucleotide CPP conjugates.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 182,327, filed April 30, 2021, and U.S. Provisional Patent Application No. 63 / 249,721, filed September 29, 2021. The entire teachings of the above applications are incorporated by reference in their entireties.

[0002] References to sequence listings submitted electronically via EFS-WEB The contents of the sequence listing submitted electronically as an ASCII text file (Name: 4140_0510002_Seqlisting_ST25, Size: 223,634 bytes, and Creation Date: April 12, 2022) are incorporated by reference in their entirety into this specification.

[0003] Field of Disclosure The present disclosure relates to a new dosing regimen for treating Duchenne muscular dystrophy (DMD) in patients with DMD using antisense oligonucleotide conjugates that induce exon skipping in the human dystrophin gene. It also provides a method for reducing toxicity associated with treatment involving antisense oligonucleotide CPP conjugates. [Background technology]

[0004] Background of the disclosure Dystrophin is a key structural protein that protects muscles from repetitive exercise-induced damage affecting skeletal, diaphragm, and cardiac muscles. Duchenne muscular dystrophy is a rare, severe, and life-threatening X-linked recessive neuromuscular degenerative disease caused by mutations in the dystrophin gene. These mutations disrupt the reading frame of the dystrophin messenger ribonucleic acid (mRNA) and prevent the translation of functional dystrophin protein. Mutations in any exon that change the reading frame of an exon or introduce a stop codon, or are characterized by the removal of an entire out-of-frame exon or the duplication of one or more exons, can prevent the production of functional dystrophin and result in DMD. The absence of dystrophin protein is the direct cause of the disease, and patients undergo a predictable disease course that begins in early childhood with an increasingly progressive deterioration of skeletal muscle function, leading to premature death, usually before the age of 30.

[0005] Duchenne muscular dystrophy (DMD) is caused by defective expression of protein dystrophin.The gene that codes for the protein contains 79 exons spread over more than 2 million nucleotides of DNA.The mutation of any exon, which changes the reading frame of the exon, introduces a stop codon, or is characterized by the removal of the entire out-of-frame exon or the duplication of one or more exons, can prevent the production of functional dystrophin and lead to DMD.

[0006] Mutations, typically deletions of one or more exons, have been shown to result in a less severe form of muscular dystrophy, Becker muscular dystrophy (BMD), if they result in the correct reading frame along the entire dystrophin transcript, and thus prevent premature termination of translation of the mRNA into protein. If the joining of upstream and downstream exons in the processing of the mutated dystrophin pre-mRNA maintains the correct reading frame of the gene, the result is an mRNA that codes for a protein with a short internal deletion that retains some activity, resulting in the BMD phenotype.

[0007] For many years, it has been known that the deletion of exon(s) that does not change the reading frame of dystrophin protein causes BMD phenotype, while the deletion of exon that causes frame shift causes DMD (Monaco, Bertelson et al. 1988). In general, dystrophin mutations, including point mutations and exon deletions that change the reading frame and thus interrupt proper protein translation, result in DMD. It should also be noted that some BMD and DMD patients have exon deletions that span multiple exons.

[0008] Antisense oligonucleotides, such as splice switch oligonucleotides (SSOs), have been successfully used in the treatment of DMD to induce alternative splicing of pre-mRNA by steric blocking of spliceosomes.SSOs are specifically designed to target specific regions of pre-mRNA, typically exons, to induce skipping of DMD gene mutations, thereby restoring these out-of-frame mutations in frame and allowing the production of internally truncated but functional dystrophin protein.Such antisense oligomers are known to target completely within exons (so-called exon internal sequences) or at splice donor or splice acceptor junctions crossing from exons to parts of introns.

[0009] For example, eteplirsen is a phosphorodiamidate morpholino oligomer (PMO) designed to skip exon 51 of the human dystrophin gene in patients with DMD, where exon 51 skipping is suitable to restore the reading frame and generate a functional truncated form of the dystrophin protein. In 2016, the US Food and Drug Administration (FDA) approved Exondys 51® (eteplirsen) for the treatment of DMD in patients with confirmed mutations of the DMD gene that are suitable for exon 51 skipping. As another example, golodyrsen (Vyondys 53®), also an antisense oligonucleotide of the PMO subclass, was recently approved for the treatment of DMD in patients with confirmed mutations of the DMD gene that are suitable for exon 53 skipping.

[0010] The discovery and development of antisense oligomers (e.g., PPMOs) conjugated to cell membrane-permeable peptides for DMD has also been an area of ​​research (see, e.g., U.S. Patent No. 10,888,578, U.S. Application No. 16 / 469,104, U.S. Application No. 16 / 001,310). Cell membrane-permeable peptides (CPPs), e.g., arginine-rich peptide transport moieties, have been shown to be effective in enhancing the penetration of antisense oligomers into cells and cause exon skipping in different muscle groups in animal models. However, to date, the relationship between human systemic exposure to dose levels of antisense oligomer conjugates (e.g., PPMOs) has not been established.

[0011] Magnesium deficiency (or hypomagnesemia) is an electrolyte disorder in which there are low levels of magnesium in the body. Causes of hypomagnesemia can include exposure to certain drugs. Hypomagnesemia can result in multiple symptoms. Symptoms include tremors, loss of coordination, muscle spasms, loss of appetite, personality changes, and nystagmus. Complications can include seizures or cardiac arrest, such as from torsades de pointes. Those with low magnesium often also have low potassium.

[0012] The diagnosis of hypomagnesemia is typically based on finding low blood magnesium levels. Normal magnesium levels can vary from about 0.6 to about 1.1 mmol / L (about 1.5 to about 2.7 mg / dL), with levels below about 0.6 mmol / L (about 1.5 mg / dL) usually defining hypomagnesemia. The effect of antisense oligomer conjugates such as PPMO on magnesium levels in patients, including DMD patients receiving PPMO therapy, is not known. Thus, despite the success achieved in preclinical models with antisense oligomers conjugated to cell membrane-permeable peptides, there remains a need for safe and effective administration paradigms for treating DMD and BMD with such conjugates in human patients. [Prior art documents] [Patent documents]

[0013] [Patent Document 1] U.S. Pat. No. 10,888,578 Summary of the Invention

[0014] In some aspects, the disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) having a mutation amenable to exon skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate of Formula (I): [ka] or a pharma- ceutically acceptable salt thereof to a patient; During the ceremony, n is 1 to 40; each Nu is a nucleobase that, taken together, forms a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA; T' is [ka] is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen and R 1 is C1-C6 alkyl, The conjugate is administered at a dose effective to provide a mean area under the curve (AUC) of the conjugate or a pharma- ceutically acceptable salt thereof of about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0015] In some embodiments, R 100 is a CPP, the CPP being attached to the 3' end of the oligomer by an amide bond at the CPP carboxy terminus. 100 is a substituent "Z," which is defined as the combination of a CPP and a linker that bridges the CPP at its carboxy terminus to the 3' end of the oligonucleotide.

[0016] In some aspects, the disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) with a mutation amenable to exon 51 skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate of Formula (I): [ka] or a pharma- ceutically acceptable salt thereof to a patient; n is 1 to 40; each Nu is a nucleobase that, taken together, forms a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA; T' is [ka] is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen and R 1is C1-C6 alkyl, The conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 300 mg to about 1500 mg of the .6HCl salt of the conjugate to achieve a mean AUC of about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0017] In some embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 130 The conjugate or pharmacologic acceptable salt thereof is administered at a dose equivalent to about 100 mg, about 500 mg to about 1000 mg, about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1300 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the .6HCl salt of the conjugate. In some embodiments, the conjugate or pharmacologic acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL. In some embodiments, the conjugate or pharmacologic acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 120 to about 240 ug·h / mL.

[0018] In certain embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 200 to about 500 ug·h / mL.

[0019] In certain embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the .6HCl salt of the conjugate.

[0020] In some embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the .6HCl salt of the conjugate.

[0021] In some embodiments, the cell membrane permeable peptide is selected from RXRRXRRXRRXR- (SEQ ID NO: 18), RFFRFFRFFR- (SEQ ID NO: 19), RXRRXRRXRRXRXB- (SEQ ID NO: 20), RFFRFFRFFRG- (SEQ ID NO: 21), RRRRRRG- (SEQ ID NO: 22), RRRRRR- (SEQ ID NO: 23), RRRRRG- (SEQ ID NO: 24), or RRRRR- (SEQ ID NO: 25), wherein R is arginine, X is 6-aminohexanoic acid, B is β-alanine, F is phenylalanine, and G is glycine.

[0022] In another embodiment, the cell membrane penetrating peptide is selected from RRRRRRG- (SEQ ID NO: 22), RRRRRR- (SEQ ID NO: 23), RRRRRG- (SEQ ID NO: 24), or RRRRR- (SEQ ID NO: 25), where R is arginine and G is glycine. In one embodiment, the cell membrane penetrating peptide is RRRRRRG- (SEQ ID NO: 22), where R is arginine and G is glycine.

[0023] In some embodiments, the antisense oligomer of the antisense oligomer conjugate has n+2 base pairs, where n in formula (I) is 1 to 40, optionally 13 to 38, optionally 13 to 28, optionally 13 to 23, or optionally 13 to 18. In other words, the oligomer is 15 to 40, 15 to 35, 15 to 30, 15 to 25, or 15 to 20 nucleotides in length.

[0024] In some embodiments, the antisense oligomer conjugate causes exon skipping in the human dystrophin gene. In some embodiments, the exon is selected from exons 44, 45, 50, 51, 52, or 53. In certain embodiments, the exon is selected from exons 45, 51, or 53.

[0025] In certain aspects, the disclosure provides a method of treating a patient with DMD having a mutation amenable to exon skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate of formula (III): [ka] or a pharma- ceutically acceptable salt thereof to a patient; wherein each Nu is a nucleobase that, together, forms a targeting sequence that is complementary to an exon annealing site in the dystrophin pre-mRNA.

[0026] In some embodiments, the disclosure provides a method of treating a patient with DMD having a mutation amenable to exon 51 skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate having formula (IV): [ka] or a pharma- ceutically acceptable salt thereof to a patient.

[0027] In other embodiments, the antisense oligomer conjugate has the formula (IVA): [ka]

[0028] In some embodiments, the antisense oligomer conjugate has formula (IVB): [ka] or a pharma- ceutically acceptable salt thereof.

[0029] In some embodiments, the antisense oligomer conjugate of formula (IVB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0030] In yet another embodiment, the disclosure provides a method of treating a patient with DMD having a mutation amenable to exon 45 skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate having formula (VI): [ka] or a pharma- ceutically acceptable salt thereof to a patient.

[0031] In some embodiments, the antisense oligomer conjugate has formula (VIA): [ka]

[0032] In some embodiments, the antisense oligomer conjugate has formula (VIB): [ka] or a pharma- ceutically acceptable salt thereof.

[0033] In some aspects, the antisense oligomer conjugate of formula (VIB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6 HCl salt.

[0034] In some aspects, the disclosure provides a method of treating a patient with DMD having a mutation amenable to exon 53 skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate having formula (VIII): [ka] or a pharma- ceutically acceptable salt thereof to a patient.

[0035] In other embodiments, the antisense oligomer conjugate has formula (VIIIA): [ka]

[0036] In other embodiments, the antisense oligomer conjugate has formula (VIIIB): [ka] or a pharma- ceutically acceptable salt thereof.

[0037] In some embodiments, the antisense oligomer conjugate of formula (VIIIB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0038] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg to about 700 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 10 kg to about 25 kg.

[0039] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg of the .6HCl salt of the conjugate.

[0040] In other aspects, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 900 mg to about 1200 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 10 kg to about 25 kg.

[0041] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 10 kg and about 25 kg at a dose equivalent to about 1100 mg of the .6HCl salt of the conjugate.

[0042] In another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing at least about 18 kg but less than about 50 kg at a dose equivalent to about 750 mg of the .6HCl salt of the conjugate.

[0043] In another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing at least about 18 kg but less than about 50 kg at a dose equivalent to about 1250 mg of the .6HCl salt of the conjugate.

[0044] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 600 mg to about 800 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 25 kg to about 50 kg.

[0045] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 25 kg and about 50 kg at a dose equivalent to about 650 mg of the .6HCl salt of the conjugate.

[0046] In other aspects, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1000 mg to about 1300 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 25 kg to about 50 kg.

[0047] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 25 kg and about 50 kg at a dose equivalent to about 1200 mg of the .6HCl salt of the conjugate.

[0048] In other aspects, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 600 mg to about 900 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0049] In another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 700 mg to about 900 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0050] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 50 kg and about 100 kg at a dose equivalent to about 750 mg of the .6HCl salt of the conjugate.

[0051] In yet another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1200 mg to about 1600 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0052] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1400 mg of the .6HCl salt of the conjugate.

[0053] In yet another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 850 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0054] In yet another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1350 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0055] In other aspects, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 600 mg to about 900 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 65 kg to about 150 kg.

[0056] In another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 700 mg to about 900 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 65 kg to about 150 kg.

[0057] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 65 kg and about 150 kg at a dose equivalent to about 750 mg of the .6HCl salt of the conjugate.

[0058] In yet another embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1200 mg to about 1600 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 65 kg to about 150 kg.

[0059] In one embodiment, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing between about 65 kg and about 150 kg at a dose equivalent to about 1400 mg of the .6HCl salt of the conjugate.

[0060] The present disclosure provides a method of treating a patient having Duchenne muscular dystrophy (DMD) with a mutation amenable to exon 51 skipping in a patient in need thereof, the method comprising administering to a patient an antisense oligomer conjugate of formula (I): [ka] or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that, taken together, forms a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA; T' is [ka] is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen and R 1 is C1-C6 alkyl, Schedule below: i) about 300 mg to about 700 mg or about 900 mg to about 1,200 mg once every 4 weeks for a patient weighing about 10 kg to about 25 kg; ii) about 600 mg to about 800 mg or about 1000 mg to about 1300 mg once every 4 weeks for a patient weighing about 25 kg to about 50 kg, or iii) about 700 mg to about 900 mg or about 1200 mg to about 1500 mg once every 4 weeks for a patient weighing about 50 kg to about 100 kg; to a patient at a dose equivalent to that of the .6HCl salt of the conjugate to achieve a mean AUC that is about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL, respectively.

[0061] In some embodiments, the conjugate is administered on the following schedule: iv) about 300 mg to about 700 mg or about 900 mg to about 1,200 mg once every 4 weeks for a patient weighing about 10 kg to about 25 kg; v) about 500 mg to about 800 mg or about 1000 mg to about 1300 mg once every 4 weeks for a patient weighing about 25 kg to about 50 kg, or vi) about 700 mg to about 900 mg or about 1200 mg to about 1500 mg once every 4 weeks for a patient weighing about 50 kg to about 100 kg; to a patient at a dose equivalent to that of the .6HCl salt of the conjugate to achieve a mean AUC that is about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL, respectively.

[0062] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg or about 1100 mg of the .6HCl salt of the conjugate.

[0063] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 750 mg or about 1250 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing between about 18 kg and about 50 kg.

[0064] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 650 mg or about 1200 mg of the .6HCl salt of the conjugate.

[0065] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 750 mg or about 1400 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0066] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 850 mg or about 1350 mg of the .6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0067] In one aspect, the disclosure provides a method of treating a patient in need of treatment, the method comprising administering to the patient an antisense oligomer conjugate and further comprising administering to the patient a magnesium supplement.

[0068] In one particular aspect, the disclosure also provides a method of treating a patient having Duchenne muscular dystrophy (DMD) with a mutation amenable to exon skipping in a patient in need thereof, the method comprising administering to the patient an antisense oligomer conjugate of Formula (I) as described herein, and further comprising administering to the patient a magnesium supplement.

[0069] The disclosure also provides a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, the method comprising administering to the patient: i) An antisense oligomeric conjugate of formula (I): [ka] or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that, taken together, forms a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA; T' is [ka] is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen and R 1is C1-C6 alkyl; ii) magnesium supplements,

[0070] In some embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 130 The .6HCl salt of the conjugate is administered at a dose equivalent to about 0 mg, about 500 mg to about 1000 mg, about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg.

[0071] In some embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the .6HCl salt of the conjugate.

[0072] In some embodiments, the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the .6HCl salt of the conjugate.

[0073] In certain embodiments, the cell membrane permeable peptide is selected from RXRRXRRXRRXR (SEQ ID NO: 18), RFFRFFRFFR (SEQ ID NO: 19), RXRRXRRXRRXRXB (SEQ ID NO: 20), RFFRFFRFFRG (SEQ ID NO: 21), RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), where R is arginine, X is 6-aminohexanoic acid, B is β-alanine, F is phenylalanine, and G is glycine. In certain embodiments, the cell membrane permeable peptide is selected from RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), where R is arginine, and G is glycine. In one particular embodiment, the cell membrane penetrating peptide is RRRRRRG (SEQ ID NO: 22), where R is arginine and G is glycine.

[0074] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof causes exon skipping in the human dystrophin gene. In some embodiments, the exon is selected from exons 44, 45, 50, 51, 52, or 53. In certain embodiments, the exon is selected from exons 45, 51, or 53.

[0075] In some embodiments, the magnesium supplement is administered in an amount effective to provide normal serum magnesium levels. In certain embodiments, the magnesium supplement is administered in a dose equivalent to about 100 to about 2400 mg of magnesium oxide per day. In other embodiments, the magnesium supplement is administered in a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0076] In some embodiments, the disclosed methods further comprise measuring the patient's serum magnesium level at two or more weeks after the administration, hi some additional embodiments, the disclosed methods comprise administering a second dose of magnesium supplement with the dose based on the measured serum magnesium level.

[0077] In some embodiments, the magnesium supplement is selected from magnesium oxide, magnesium citrate, magnesium carbonate, magnesium hydrogen phosphate, magnesium glycerophosphate, magnesium trisilicate, magnesium hydroxide, magnesium carbonate hydroxide, magnesium acetate, magnesium citrate, magnesium lactate, magnesium gluconate, magnesium chloride, magnesium aspartate, magnesium caprylate, magnesium ascorbate, magnesium taurate, magnesium malate, and magnesium diglycinate, magnesium pidulate, or magnesium sulfate. In one embodiment, the magnesium supplement is derived from magnesium oxide. [Brief description of the drawings]

[0078] [Figure 1] 1 shows the difference in initial body weight of animals in two 12 week old non-human primate (NHP) studies. [Diagram 2] 1 shows plasma exposure versus body weight in NHPs following administration of PPMO-1. [Diagram 3] Figure 1 shows area under the curve (AUC) and peak plasma concentration (Cmax) versus body weight in NHP following administration of PPMO-1. Higher body weight is associated with higher AUC and Cmax. [Figure 4] FIG. 1 is a graph showing peak plasma concentrations (Cmax) of PPMO-1 at 20 and 30 mg / kg in human DMD patients. [Diagram 5] The correlation between peak plasma concentrations (Cmax) and body weight in DMD patients receiving either 20 mg / kg PPMO-1 (bottom) or 30 mg / kg PPMO-1 (top). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0079] The present disclosure relates to novel methods of administration for treating muscular dystrophies, such as DMD and BMD, by administering antisense oligomer conjugates, or pharma- ceutically acceptable salts thereof, specifically designed to induce exon skipping in the human dystrophin gene.

[0080] The present disclosure also provides therapeutically effective dosing regimens that minimize undesirable side effects associated with administration of antisense oligomer conjugates. Specifically, the present disclosure provides methods for reducing the frequency and severity of hypomagnesemia in patients being treated with administration of antisense oligomer conjugates.

[0081] definition "About" means a quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length that varies by up to 30%, 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the reference quantity, level, value, number, frequency, percentage, dimension, size, amount, weight, or length.

[0082] As used herein, the term "alkyl" refers to a saturated straight chain or branched hydrocarbon, unless otherwise specified. In certain embodiments, an alkyl group is a primary, secondary, or tertiary hydrocarbon. In certain embodiments, an alkyl group is a group having 1 to 10 carbon atoms, i.e., C1-C 10and alkyl. In certain embodiments, the alkyl group comprises 1 to 6 carbon atoms, i.e., C1-C6 alkyl. In certain embodiments, the alkyl group is selected from the group consisting of methyl, CF3, CCl3, CFCl2, CF2Cl, ethyl, CH2CF3, CF2CF3, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, and 2,3-dimethylbutyl. The term includes both substituted and unsubstituted alkyl groups, including halogenated alkyl groups. In certain embodiments, the alkyl group is a fluorinated alkyl group. Non-limiting examples of moieties with which an alkyl group may be substituted are selected from the group consisting of halogen (fluoro, chloro, bromo, or iodo), hydroxyl, amino, alkylamino, arylamino, alkoxy, aryloxy, nitro, cyano, sulfonic acid, sulfate, phosphonic acid, phosphate, or phosphonate, either unprotected or optionally protected, as known to those of skill in the art, e.g., as taught in Greene, et al., Protective Groups in Organic Synthesis, John Wiley and Sons, Second Edition, 1991, which is incorporated herein by reference.

[0083] "Suitable for exon skipping", as used herein with respect to a subject or patient, is intended to include subjects and patients with one or more mutations in the dystrophin gene, in which the absence of skipping of certain exons in the dystrophin pre-mRNA results in an out-of-frame reading frame, thereby preventing translation of the pre-mRNA and rendering the subject or patient unable to produce functional or semi-functional dystrophin. It is well within the understanding of those skilled in the art to determine whether a patient has a mutation in the dystrophin gene that is suitable for exon skipping (see, for example, Aartsma-Rus et al. (2009) Hum Mutat. 30:293-299, Gurvich et al., Hum Mutat. 2009; 30(4)633-640, and Fletcher et al. (2010) Molecular Therapy 18(6)1218-1223).

[0084] The terms "oligomer" and "oligonucleotide" are used interchangeably and refer to a sequence of subunits linked by intersubunit bonds. In certain instances, the term "oligomer" is used in connection with "antisense oligomers." For "antisense oligomers," each subunit consists of (i) a ribose sugar or derivative thereof, and (ii) a nucleic acid base bound thereto, whereby the order of base-pairing moieties forms a base sequence that is complementary to a target sequence in a nucleic acid (typically RNA) by Watson-Crick base pairing to form a nucleic acid:oligomer heteroduplex within the target sequence, provided that either the subunits, the intersubunit linkages, or both, are not naturally occurring. In certain embodiments, the antisense oligomer is a PMO.

[0085] The terms "complementary" and "complementarity" refer to two or more oligomers (i.e., each containing a nucleobase sequence) related to each other by Watson-Crick base pairing rules. For example, the nucleobase sequence "TGA(5'→3')" is complementary to the nucleobase sequence "ACT(3'→5')". Complementarity can be "partial", where less than all of the nucleobases of a given nucleobase sequence are matched with another nucleobase sequence according to the base pairing rules. For example, in some embodiments, the complementarity between a given nucleobase sequence and another nucleobase sequence can be about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. Alternatively, continuing the example, there can be "complete" or "perfect" (100%) complementarity between a given nucleobase sequence and another nucleobase sequence. The degree of complementarity between nucleobase sequences has a significant effect on the efficiency and strength of hybridization between the sequences.

[0086] The terms "effective amount" and "therapeutically effective amount" are used interchangeably herein and refer to an amount of a therapeutic compound, such as an antisense oligomer conjugate, administered to a mammalian subject, either as a single dose or as part of a series of doses, that is effective to produce a desired therapeutic effect. For antisense oligomer conjugates, this effect is typically produced by inhibiting translation or natural splice processing of a selected target sequence, or by producing a clinically significant amount of dystrophin (statistical significance).

[0087] "Enhance" or "enhancing", or "increase" or "increasing", or "stimulate" or "stimulating" generally refers to the ability of one or more antisense oligomer conjugates or pharmaceutical compositions to produce or cause a greater physiological response (i.e., a downstream effect) in a cell or subject compared to the response caused by either no antisense oligomer conjugate, or a control compound. A greater physiological response can include increased expression of a functional form of dystrophin protein, or increased dystrophin-related biological activity in muscle tissue, among other responses apparent from understanding in the art and described herein. Increases in muscle function may also be measured, including increases or improvements in muscle function by up to about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. The percentage of muscle fibers expressing functional dystrophin can also be measured, including increasing dystrophin expression by about 1%, 2%, 5%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of muscle fibers. For example, it has been shown that when 25-30% of fibers express dystrophin, an improvement in muscle function of about 40% can occur (see, e.g., DelloRusso et al., Proc Natl Acad Sci USA 99:12979-12984, 2002). An "increased" or "enhanced" amount is typically a "statistically significant" amount and can include an increase that is 1.1, 1.2, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 30, 40, 50-fold or more (e.g., 500, 1000-fold, including all integers and decimals therebetween and greater than 1, e.g., 1.5, 1.6, 1.7, 1.8, etc.) over the amount produced by no antisense oligomer conjugate (absence of agent) or a control compound.

[0088] As used herein, terms such as "function" and "functionality" refer to biological, enzymatic, or therapeutic functions.

[0089] A "functional" dystrophin protein generally refers to a dystrophin protein that has sufficient biological activity to reduce the progressive degradation of muscle tissue otherwise characteristic of muscular dystrophies, as compared to altered or "defective" forms of dystrophin protein typically present in certain subjects with DMD or BMD. In certain embodiments, a functional dystrophin protein may have about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% (including all integers therebetween) of the in vivo biological activity of wild-type dystrophin, as measured according to routine techniques in the art. Truncated forms of dystrophin are included, such as those forms produced after administration of certain exon-skipping antisense oligomer conjugates of the present disclosure.

[0090] The term "mismatch" refers to one or more nucleobases (whether contiguous or separated) in the oligomer nucleobase sequence that do not match the target pre-mRNA according to base pairing rules. While perfect complementarity is often desired, some embodiments may include one or more, but preferably 6, 5, 4, 3, 2, or 1 mismatches to the target pre-mRNA. Variations at any position within the oligomer are included. In certain embodiments, the antisense oligomer conjugates of the present disclosure include variations in nucleobase sequence near the internal terminal variations, when present, typically within about 6, 5, 4, 3, 2, or 1 subunit of the 5' and / or 3' end.

[0091] The terms "morpholino," "morpholino oligomer," and "PMO" have the following general structure: [ka] and as depicted in Figure 2 of Summerton, J., et al., Antisense & Nucleic Acid Drug Development, 7:187-195 (1997). The morpholino described herein includes all stereoisomers and tautomers of the above general structure. The synthesis, structure and binding properties of morpholino oligomers are described in detail in U.S. Patent Nos. 5,698,685, 5,217,866, 5,142,047, 5,034,506, 5,166,315, 5,521,063, 5,506,337, 8,076,476 and 8,299,206, all of which are incorporated herein by reference.

[0092] In some embodiments, morpholino oligos (PMOs) are conjugated with a "tail" moiety at the 5' or 3' end of the oligomer to enhance its stability and / or solubility. Exemplary tails include: [ka] Includes.

[0093] Of the exemplary tail moieties above, "TEG" or "EG3" refers to the following tail moieties: [ka]

[0094] Of the exemplary tail moieties above, "GT" refers to the following tail moieties: [ka]

[0095] Useful morpholino oligomers are described in more detail below.

[0096] The terms "antisense oligomer CPP conjugate," "antisense oligomer conjugate," and "PPMO" refer to a conjugate in which a PMO is conjugated to a cell membrane penetrating peptide. CPPs are described more fully below.

[0097] As seen in the table below, non-limiting examples of CPPs for use herein include -(RXR)4-R a , -R-(FFR)3-R a , -BX-(RXR)4-R a , -BXR-(FFR)3-R a , -GLY-R-(FFR)3-R a , -GLY-R5-R a and -R5-R a , -GLY-R6-R a and -R6-R a R a is selected from H, acyl, acetyl, benzoyl, and stearoyl, R is arginine, X is 6-aminohexanoic acid, B is β-alanine, F is phenylalanine, and GLY (or G) is glycine. CPP "R6" is intended to indicate a peptide of six arginine residues linked together via amide bonds (a single substituent, e.g., R 6 In some embodiments, R a is H or acyl. In some embodiments, R a is acetyl. Exemplary CPPs are provided in Table 1 below.

[0098] In one embodiment, the CPP is selected from one of the following sequences: [Table 19]

[0099] CPPs, their synthesis, and methods of conjugating to oligomers are further described in U.S. Patent Nos. 9,161,948 and 10,888,578, U.S. Application Publication No. 2012 / 0289457, and International Patent Application Publication Nos. 2004 / 097017, 2009 / 005793, and 2012 / 150960, the disclosures of which are incorporated herein by reference in their entireties. A more comprehensive list of CPPs is provided below.

[0100] The terms "nucleobase" (Nu), "base-pairing moiety," or "base" are used interchangeably to refer to the purine or pyrimidine bases found in naturally occurring or "natural" DNA or RNA (e.g., uracil, thymine, adenine, cytosine, and guanine), as well as analogs of these naturally occurring purines and pyrimidines. These analogs may confer improved properties, such as binding affinity, to the oligomer. Exemplary analogs include hypoxanthine (the base component of inosine), 2,6-diaminopurine, 5-methylcytosine, C5-propynyl-modified pyrimidines, 10-(9-(aminoethoxy)phenoxazinyl) (G-clamp), and the like.

[0101] Further examples of base pairing moieties include, but are not limited to, uracil, thymine, adenine, cytosine, guanine, and hypoxanthine (inosine) (with their respective amino groups protected by acyl protecting groups), pyrimidine analogs such as 2-fluorouracil, 2-fluorocytosine, 5-bromouracil, 5-iodouracil, 2,6-diaminopurine, azacytosine, pseudoisocytosine and pseudouracil, and other modified nucleobases such as 8-substituted purines, xanthines, or hypoxanthines (the latter two are natural decomposition products). Also contemplated are modified nucleobases disclosed in Chiu and Rana, RNA, 2003, 9, 1034-1048; Limbach et al. Nucleic Acids Research, 1994, 22, 2183-2196; and Revankar and Rao, Comprehensive Natural Products Chemistry, vol. 7, 313, the contents of which are incorporated herein by reference.

[0102] Further examples of base pairing moieties include, but are not limited to, extended size nucleobases with one or more additional benzene rings. Nucleobase substitutions described in the Glen Research catalog (www.glenresearch.com), Krueger AT et al., Acc. Chem. Res., 2007, 40, 141-150, Kool, ET, Acc. Chem. Res., 2002, 35, 936-943, Benner SA, et al., Nat. Rev. Genet., 2005, 6, 553-543, Romesberg, FE, et al., Curr. Opin. Chem. Biol., 2003, 7, 723-733, and Hirao, I., Curr. Opin. Chem. Biol., 2006, 10, 622-627, the contents of which are incorporated herein by reference, are contemplated as useful in the antisense oligomer conjugates described herein. Examples of extended size nucleobases include those shown below, as well as tautomers thereof. [ka]

[0103] The term "exposure" refers to dose (PPMO input into the body) and various measures of acute or integrated PPMO concentration in plasma and other biological fluids (e.g., Cmax, Cmin, Css, AUC). The term "response" refers to a direct measure of the pharmacological effect of the drug. Responses include a wide range of endpoints or biomarkers, ranging from potential or acceptable surrogates (e.g., effects on blood pressure, magnesium levels, or cardiac output) to short-term or long-term clinical effects related to efficacy and safety.

[0104] As used herein, the phrases "parenteral administration" and "administered parenterally" refer to modes of administration other than enteral and topical administration, usually by injection, and include, but are not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0105] For clarity, the structures of the present disclosure are continuous from 5' to 3' and include various diagrammatic breaks labeled "Break A," "Break B," and "Break C" for the convenience of depicting the entire structure in a compact form. As will be understood by those skilled in the art, for example, each of the "Break A" designations indicates the diagrammatic continuation of the structure at these points. Those skilled in the art will understand that the same is true for each of the instances of "Break B" and "Break C" in the above structures. However, none of the diagrammatic breaks are intended to indicate an actual discontinuity in the above structures, and those skilled in the art would not understand them to be so meant.

[0106] As used herein, a set of brackets used within a structural formula indicates that the structural feature between the brackets is repeated. In some embodiments, the brackets used may be "[" and "]", and in certain embodiments, the brackets used to indicate a repeating structural feature may be "(" and ")". In some embodiments, the number of repeating repetitions of the structural feature between the brackets is the number shown outside the brackets, e.g., 2, 3, 4, 5, 6, 7, etc. In various embodiments, the number of repeating repetitions of the structural feature between the brackets is indicated by a variable shown outside the brackets, such as "n".

[0107] As used herein, a straight or squiggly bond drawn to a chiral carbon or phosphorus atom in a structural formula indicates that the stereochemistry of the chiral carbon or phosphorus is undefined and is intended to include all forms of the chiral center. Examples of such diagrams are shown below. [ka]

[0108] Management of hypomagnesemia The present disclosure provides a method for reducing the severity and frequency of hypomagnesemia in DMD patients. The incidence of hypomagnesemia in DMD patients is generally equal to or less than that in healthy individuals. Unexpectedly, the present inventors have observed that patients treated with antisense oligomeric CPP conjugates (e.g., PPMO), higher exposure (AUC value) correlates with the severity and frequency of hypomagnesemia (magnesium deficiency). Thus, the methods described herein can result in a reduction in toxicity associated with abnormal magnesium levels associated with the administration of antisense oligomeric CPP conjugates (e.g., PPMO) in human patients.

[0109] In some aspects, the disclosure provides a method of treating hypomagnesemia associated with administering an antisense oligomeric CPP conjugate (e.g., PPMO) to a patient to effect exon skipping by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate. In some aspects, the patient is a DMD patient. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (I) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (IVB) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIB) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIIIB) or a pharmaceutically acceptable salt thereof.

[0110] In some aspects, the disclosure provides a method of preventing hypomagnesemia associated with administering an antisense oligomeric CPP conjugate (e.g., PPMO) to a patient to effect exon skipping by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate. In some aspects, the patient is a DMD patient. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (I) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (IVB) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIB) or a pharmaceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIIIB) or a pharmaceutically acceptable salt thereof.

[0111] In some embodiments, the present disclosure provides a method for maintaining normal serum magnesium levels in a patient to whom an antisense oligomeric conjugate (e.g., PPMO) is administered by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate. In some embodiments, the patient is a DMD patient. In some embodiments, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (IVB) or a pharmaceutically acceptable salt thereof. In some embodiments, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIB) or a pharmaceutically acceptable salt thereof. In some embodiments, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIIIB) or a pharmaceutically acceptable salt thereof.

[0112] Hypomagnesemia in patients (e.g., DMD patients) can be diagnosed by physical examination, symptoms, medical history, and laboratory tests to test for magnesium body content. Generally speaking, means for detecting magnesium body content include detecting blood levels of magnesium in the patient's plasma or serum (the abnormalities of which generally indicate a disorder of magnesium metabolism and are usually the starting point for a set of further specific tests), detecting magnesium levels in urine (which gives a measure of magnesium excretion via urine, usually related to protein intake, with a fairly constant urinary Mg / urea ratio), detecting magnesium levels in spinal fluid, detecting erythrocyte magnesium (which indicates the amount of Mg contained in the bone marrow when erythropoiesis is occurring and possible, thus an indirect marrow survey for magnesium), detecting lymphocyte magnesium, 25These include nuclear magnetic resonance with Mg, which demonstrates the intracellular distribution of magnesium and any modifications in its different chemical and physical structures, and detection of magnesium content in the patient's bone, muscle, or any other tissue or organ of interest.

[0113] Hypomagnesemia is defined according to the CTCAE (Common Terminology Criteria for Adverse Events) or reduced magnesium levels: [Table 20]

[0114] Severe hypomagnesemia usually results in a concentration below 0.9 mg / dL (<0.4 mmol / L).

[0115] One aspect of the present disclosure includes treating a patient diagnosed with hypomagnesemia by administering a magnesium supplement to the patient. Throughout this specification, in the context of magnesium-containing products, magnesium supplements, magnesium formulations, and / or magnesium therapy, the term "magnesium" refers to Mg, either in the form of a free ion in a salt or in the form of a complex. +2 Refers to ions.

[0116] In certain embodiments, magnesium supplements are administered prophylactically without first testing for hypomagnesemia. One embodiment of the present disclosure includes treating a patient with Duchenne muscular dystrophy (DMD) with an antisense oligomer CPP conjugate, the method comprising administering to the patient (i) an antisense oligonucleotide CPP conjugate comprising a morpholino phosphorodiamidate antisense oligomer that is (a) 100% complementary to a portion of an exon of human dystrophin pre-mRNA, and (b) 15-50 nucleotides in length (the antisense oligonucleotide is administered intravenously), and a magnesium supplement. In another embodiment, the oligomer is 15-40, 15-35, 15-30, 15-25, or 15-20 nucleotides in length. In some embodiments, the oligo is 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, or 15 nucleotides in length. In one aspect, the oligo is 30 nucleotides in length. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (I) or a pharma- ceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (IVB) or a pharma- ceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIB) or a pharma- ceutically acceptable salt thereof. In some aspects, the antisense oligomeric CPP conjugate is an antisense oligomeric CPP conjugate of formula (VIIIB) or a pharma- ceutically acceptable salt thereof.

[0117] In some embodiments, the magnesium supplement is administered orally to the patient. Examples of magnesium-containing products for oral administration include magnesium oxide, magnesium citrate, magnesium carbonate, magnesium hydrogen phosphate, magnesium glycerophosphate, magnesium trisilicate, magnesium hydroxide, magnesium carbonate hydroxide, magnesium acetate, magnesium citrate, magnesium lactate, magnesium gluconate, magnesium chloride, magnesium aspartate, magnesium caprylate, magnesium ascorbate, magnesium taurate, magnesium malate, and magnesium diglycinate, magnesium pidulate, or magnesium sulfate. In some embodiments, the magnesium supplement is magnesium oxide.

[0118] In some embodiments, the magnesium supplement is administered to the patient intravenously. Intravenously administered magnesium includes, for example, magnesium sulfate.

[0119] As with many metal-containing complexes, different magnesium compounds have different mechanisms of uptake in vivo. Furthermore, after administration, including ingestion, transition metal complexes may undergo any number of reactions in vivo that affect bioavailability and resulting therapeutic effects. As an example, magnesium citrate is soluble in gastric juice and therefore readily available for passive absorption in the upper gastrointestinal tract. Magnesium acetate, magnesium ascorbate, and magnesium lactate are soluble in gastric juice and share the upper gastrointestinal passive absorption capacity of magnesium citrate. Ascorbic acid radicals serve as a source of vitamin C by conversion to ascorbic acid after exposure to hydrochloric acid in gastric juice, while magnesium ions are converted to soluble magnesium chloride. The sufficient water solubility of magnesium acetate, magnesium ascorbate, magnesium citrate, and magnesium lactate provides a diffusion gradient of magnesium in the upper small intestine over which some passive absorption of magnesium occurs. Magnesium oxide is converted to magnesium chloride in the stomach, providing the advantage of a high ionic magnesium content, as 60% by weight of the magnesium oxide molecule is elemental magnesium. Magnesium diglycinate represents a form of magnesium that is partially absorbed as an intact dipeptide in the proximal small intestine via the dipeptide transport pathway, thus providing a third absorption mechanism for magnesium.

[0120] Various dosage amounts of magnesium are contemplated by the present disclosure. Unless expressly stated otherwise, dosage amounts referred to herein refer to amounts equivalent to magnesium oxide. Doses may range from about 5 mg to about 1000 mg or more. In some embodiments, doses may range from about 5 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, or about 50 mg. In some embodiments, the dose may be in the range of about 15 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, or about 50 mg. In some embodiments, the dose may be in the range of about 30 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, or about 50 mg. In some embodiments, the dose may be in the range of about 60 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, about 250 mg, about 200 mg, about 150 mg, about 100 mg, or about 50 mg. In some embodiments, the dose may be in the range of about 100 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, about 250 mg, or about 200 mg. In some embodiments, the dose can be in the range of about 200 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, about 350 mg, about 300 mg, to about 250 mg.In some embodiments, the dose may be in the range of about 300 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, about 450 mg, about 400 mg, or about 350 mg. In some embodiments, the dose may be in the range of about 400 mg to about 900 mg, about 850 mg, about 800 mg, about 750 mg, about 700 mg, about 650 mg, about 600 mg, about 550 mg, about 500 mg, or about 450 mg.

[0121] The specific dosage of magnesium supplement is about 5mg, about 10mg, about 15mg, about 20mg, about 25mg, about 30mg, about 35mg, about 40mg, about 45mg, 50mg, 55mg, 60mg, 65mg, 70mg, 75mg, 80mg, 85mg, 90mg, 95mg, about 100mg, about 105mg, about 110mg, about 115mg, about 120mg, about 125mg, about 130mg, about 135mg, about 140mg, about 145mg, about 150mg, about 155mg, about 160mg, about 165mg, about 170mg, about 175mg, about 180mg, about 185mg, about 190mg, about 200mg, about 205mg, about 210mg, about 215mg, about 220mg, about 225mg, about 230mg, about 235mg, 240mg, about 245mg, about 250mg, about 255mg, about 260mg, about 265mg, about 270mg, about 275mg, about 280mg, about 285mg, about 290mg, about 295mg, about 300mg, about 305mg, about 310mg, about 315mg, about 320mg, about 325mg, about 330mg, about 335mg, about 340mg, about 345mg, about 350mg, about 355mg, about 360mg, about 365mg, about 370mg, about 375mg, about 380mg, about 385mg, about 390mg, about 395mg, about 400mg, about 405mg, about 410mg, about 415mg, about 420mg, about 425mg, about 430mg, about 435mg, about 440mg, about 445mg, about 450mg, about 455mg, about 460mg, about 465mg, about 470mg, about 475mg, about 480mg, about 485mg, about 490mg, about 495mg, about 500mg, about 505mg, about 510mg, about 515mg, about 520mg, about 525mg, about 530mg, about 535mg, about 540mg, about 545mg, about 550mg, about 555mg, about 560mg, about 565mg, about 570mg, about 575mg, about 580mg, about 585mg, about 590mg, about 595mg, about 600mg, about 605mg, about 610mg, about 615mg, about 620mg, about 625mg, about 630mg, about 635mg, about 640mg, about 645mg, about 650mg, about 655mg, 660mg, about 665mg, about 670mg, about 675mg, about 680mg, about 685mg, about 690mg, about 695mg, about 700mg, about 705mg, about 710mg, about 715mg, about 720mg, about 725mg,About 730mg, about 735mg, about 740mg, about 745mg, about 750mg, about 755mg, about 760mg, about 765mg, 770mg, about 780mg, about 785mg, about 790mg, about 795mg, About 800mg, about 805mg, about 810mg, about 815mg, about 820mg, about 825mg, about 830mg, about 835mg, about 840mg, about 845mg, 850mg, about 855mg, about 860mg, about 865 mg, 880 mg, about 885 mg, about 890 mg, about 895 mg, about 900, about 905 mg, about 910 mg, about 915 mg, about 920 mg, about 925 mg, about 930 mg, about 935 mg, about 940 mg, about 945 mg, 950 mg, about 955 mg, 960 mg, about 965 mg, about 970 mg, about 975 mg, about 980 mg, about 985 mg, about 990 mg, about 995 mg, or about 1000 mg.

[0122] In some aspects, the disclosure provides methods for treating or preventing hypomagnesemia associated with administering to a patient an antisense oligomeric CPP conjugate of Formula (IVB) or a pharma- ceutically acceptable salt thereof to achieve exon skipping by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate, wherein the magnesium supplement dose is between about 400 mg and 900 mg.

[0123] In some aspects, the disclosure provides methods for treating or preventing hypomagnesemia associated with administering to a patient an antisense oligomeric CPP conjugate of Formula (VIB) or a pharma- ceutically acceptable salt thereof to achieve exon skipping by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate, wherein the magnesium supplement dose is between about 400 mg and 900 mg.

[0124] In some aspects, the disclosure provides methods for treating or preventing hypomagnesemia associated with administering to a patient an antisense oligomeric CPP conjugate of Formula (VIIIB) or a pharma- ceutically acceptable salt thereof to achieve exon skipping by administering a magnesium supplement to the patient being treated with the antisense oligomeric CPP conjugate, wherein the magnesium supplement dose is between about 400 mg and 900 mg.

[0125] In one embodiment, the dose of magnesium supplement is equivalent to about 400 mg of magnesium oxide. In another embodiment, the dose of magnesium supplement is equivalent to about 800 mg of magnesium oxide.

[0126] Higher doses of magnesium are also contemplated. For example, the dose may be in the range of about 1000 mg to about 2400 mg, about 1000 mg to about 2000 mg, about 1000 mg to about 1500 mg, about 1050 mg to about 2400 mg, about 1050 mg to about 2000 mg, about 1050 mg to about 1500 mg, 1100 mg to about 2400 mg, about 1100 mg to about 2000 mg, about 1100 mg to about 1500 mg, 1200 mg to about 2400 mg, about 120 mg to about 2000 mg, about 12 mg to about 1500 mg, 1500 mg to about 2400 mg, or about 1500 mg to about 2000 mg. Specific doses of magnesium can include about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, about 1550 mg, about 1600 mg, about 1650 mg, about 1700 mg, about 1750 mg, about 1800 mg, about 1850 mg, about 1900 mg, about 1950 mg, about 2000 mg, about 2050 mg, about 2100 mg, about 2150 mg, about 2200 mg, about 2250 mg, about 2300 mg, about 2350 mg, or about 2400 mg.

[0127] Higher dosages may be used as a short-term regimen (e.g., less than about one month) and may be tapered to dosages at the lower end of the range. In many such cases, it will be advantageous to not go below about 40-100 mg of magnesium per day when tapering the dose. Alternatively, the appropriate dose of magnesium may be based on the condition of any subject, as assessed by a number of important variables. Alternatively, the dosage may depend on the mode of administration. Alternatively, the magnesium dosage may vary with the amount of antisense oligomer CPP conjugate. The appropriate dosage may depend on a number of factors and may be readily determined by one of skill in the art.

[0128] In some embodiments, the magnesium supplement is administered once daily. In other embodiments, the magnesium supplement is administered twice daily. In still other embodiments, the magnesium supplement is administered three times daily. More frequent daily administration of magnesium supplements, as determined by the treating physician, is also contemplated by the present disclosure.

[0129] The magnesium supplement and the antisense oligomeric CPP conjugate can be administered to an individual in any order. In one embodiment, the magnesium supplement and the antisense oligomeric CPP conjugate are administered simultaneously. For example, the magnesium supplement and the antisense oligomeric CPP conjugate can be administered within the same time period of each other. In some embodiments, the magnesium supplement and the antisense oligomeric CPP conjugate can also be administered within about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or about 24 hours of each other.

[0130] In other embodiments, the antisense oligomeric CPP conjugate may be administered once every 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, while the magnesium supplement is administered once, twice, three, four or more times per day. In other embodiments, the antisense oligomeric CPP conjugate may be administered once every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, while the magnesium supplement is administered once, twice, three, four or more times per day.

[0131] Magnesium supplementation can also be started before the start of treatment regimen with antisense oligomeric CPP conjugate.For example, a patient can start magnesium supplementation 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 days or more before the first treatment with antisense oligomeric CPP conjugate.Alternatively, magnesium supplementation can also be started after the start of treatment regimen with antisense oligomeric CPP conjugate. For example, a patient can begin magnesium supplementation 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or more days after initial treatment with an antisense oligomeric CPP conjugate.

[0132] In another embodiment, any of these magnesium supplement embodiments may be combined with one of the PPMO administration embodiments described below.

[0133] The serum magnesium level of the patient should be measured periodically during treatment with the antisense oligomer CPP conjugate. In some embodiments, the serum magnesium level of the patient is measured before administering the antisense oligomer CPP conjugate. In some embodiments, the serum magnesium level of the patient is measured 2 weeks or more after administration of the conjugate. Depending on the serum magnesium level of the patient at a given measurement, the dose of magnesium supplement can be adjusted upward or downward to maintain normal serum magnesium level. Thus, it is also contemplated by the present disclosure to administer a second dose of magnesium supplement, with the dose based on the measured serum magnesium level.

[0134] Administration of antisense oligomeric CPP conjugates (PPMOs) Previous clinical settings involving splice switching oligomers (SSOs) (e.g., PMOs such as eteplirsen, gologirsen, etc.) for the treatment of DMD have used a weight-based (mg / kg) approach, where it was unexpectedly discovered that both dystrophin expression and safety (hypomagnesemia) were related to PPMO exposure (dose).

[0135] Using a weight-based (mg / kg) PPMO dosing approach, it was discovered that higher body weight (BW) was associated with higher drug exposure, resulting in a risk of underdosing in lighter patients and overdosing in heavier patients. Thus, a weight-based dosing approach was found to result in suboptimal dystrophin expression in lighter patients and potential hypomagnesemia in heavier patients.

[0136] The present disclosure provides novel dosing regimens developed to optimize dystrophin expression and minimize concerns about hypomagnesemia in human patients.

[0137] In some embodiments, the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, is administered at a dose effective to provide a target mean AUC that maximizes dystrophin expression and minimizes hypomagnesemia in the patient.

[0138] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL, about 100 to about 190 ug·h / mL, about 100 to about 180 ug·h / mL, about 100 to about 170 ug·h / mL, about 100 to about 160 ug·h / mL, about 100 to about 150 ug·h / mL, about 100 to about 140 ug·h / mL, about 100 to about 130 ug·h / mL, about 100 to about 120 ug·h / mL, or about 100 to about 110 ug·h / mL.

[0139] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 200 ug·h / mL, about 120 to about 200 ug·h / mL, about 130 to about 200 ug·h / mL, about 140 to about 200 ug·h / mL, about 150 to about 200 ug·h / mL, about 160 to about 200 ug·h / mL, about 170 to about 200 ug·h / mL, about 180 to about 200 ug·h / mL, or about 190 to about 200 ug·h / mL.

[0140] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 190 ug·h / mL, about 120 to about 190 ug·h / mL, about 130 to about 190 ug·h / mL, about 140 to about 190 ug·h / mL, about 150 to about 190 ug·h / mL, about 160 to about 190 ug·h / mL, about 170 to about 190 ug·h / mL, or about 180 to about 190 ug·h / mL.

[0141] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 180 ug·h / mL, about 120 to about 180 ug·h / mL, about 130 to about 180 ug·h / mL, about 140 to about 180 ug·h / mL, about 150 to about 180 ug·h / mL, about 160 to about 180 ug·h / mL, or about 170 to about 180 ug·h / mL.

[0142] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 170 ug·h / mL, about 120 to about 170 ug·h / mL, about 130 to about 170 ug·h / mL, about 140 to about 170 ug·h / mL, about 150 to about 170 ug·h / mL, or about 160 to about 170 ug·h / mL.

[0143] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 160 ug·h / mL, about 120 to about 160 ug·h / mL, about 130 to about 160 ug·h / mL, about 140 to about 160 ug·h / mL, or about 150 to about 160 ug·h / mL.

[0144] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 110 to about 150 ug·h / mL, about 120 to about 150 ug·h / mL, about 130 to about 150 ug·h / mL, or about 140 to about 150 ug·h / mL.

[0145] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 120 to about 240 ug·h / mL, about 120 to about 230 ug·h / mL, about 120 to about 220 ug·h / mL, about 120 to about 210 ug·h / mL, about 120 to about 200 ug·h / mL, about 120 to about 190 ug·h / mL, about 120 to about 180 ug·h / mL, about 120 to about 170 ug·h / mL, about 120 to about 160 ug·h / mL, about 120 to about 150 ug·h / mL, about 120 to about 140 ug·h / mL, or about 120 to about 130 ug·h / mL.

[0146] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 200 ug·h / mL, about 130 to about 190 ug·h / mL, about 130 to about 180 ug·h / mL, about 130 to about 170 ug·h / mL, about 130 to about 160 ug·h / mL, about 130 to about 150 ug·h / mL, or about 130 to about 140 ug·h / mL.

[0147] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 140 to about 200 ug·h / mL, about 140 to about 190 ug·h / mL, about 140 to about 180 ug·h / mL, about 140 to about 170 ug·h / mL, about 140 to about 160 ug·h / mL, or about 140 to about 150 ug·h / mL.

[0148] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 150 to about 200 ug·h / mL, about 150 to about 190 ug·h / mL, about 150 to about 180 ug·h / mL, about 150 to about 170 ug·h / mL, or about 150 to about 160 ug·h / mL.

[0149] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 160 to about 200 ug·h / mL, about 160 to about 190 ug·h / mL, about 160 to about 180 ug·h / mL, or about 160 to about 170 ug·h / mL.

[0150] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 170 to about 200 ug·h / mL, about 170 to about 190 ug·h / mL, or about 170 to about 180 ug·h / mL.

[0151] In some embodiments, the antisense oligomeric CPP conjugate, or a pharma- ceutically acceptable salt thereof, is administered at a dose effective to provide a mean AUC that is about 130 to about 240 ug·h / mL, about 140 to about 240 ug·h / mL, about 150 to about 240 ug·h / mL, about 160 to about 240 ug·h / mL, about 170 to about 240 ug·h / mL, about 180 to about 240 ug·h / mL, about 190 to about 240 ug·h / mL, about 200 to about 240 ug·h / mL, about 210 to about 240 ug·h / mL, about 220 to about 240 ug·h / mL, or about 230 to about 240 ug·h / mL.

[0152] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 230 ug·h / mL, about 140 to about 230 ug·h / mL, about 150 to about 230 ug·h / mL, about 160 to about 230 ug·h / mL, about 170 to about 230 ug·h / mL, about 180 to about 230 ug·h / mL, about 190 to about 230 ug·h / mL, about 200 to about 230 ug·h / mL, about 210 to about 230 ug·h / mL, or about 220 to about 230 ug·h / mL.

[0153] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 220 ug·h / mL, about 140 to about 220 ug·h / mL, about 150 to about 220 ug·h / mL, about 160 to about 220 ug·h / mL, about 170 to about 220 ug·h / mL, about 180 to about 220 ug·h / mL, or about 190 to about 220 ug·h / mL, about 200 to about 220 ug·h / mL, or about 210 to about 220 ug·h / mL.

[0154] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 210 ug·h / mL, about 140 to about 210 ug·h / mL, about 150 to about 210 ug·h / mL, about 160 to about 210 ug·h / mL, about 170 to about 210 ug·h / mL, about 180 to about 210 ug·h / mL, about 190 to about 210 ug·h / mL, or about 200 to about 210 ug·h / mL.

[0155] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 200 ug·h / mL, about 140 to about 200 ug·h / mL, about 150 to about 200 ug·h / mL, about 160 to about 200 ug·h / mL, about 170 to about 200 ug·h / mL, about 180 to about 200 ug·h / mL, or about 190 to about 200 ug·h / mL.

[0156] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 190 ug·h / mL, about 140 to about 190 ug·h / mL, about 150 to about 190 ug·h / mL, or about 160 to about 190 ug·h / mL, about 170 to about 190 ug·h / mL, or about 180 to about 190 ug·h / mL.

[0157] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 180 ug·h / mL, about 140 to about 180 ug·h / mL, about 150 to about 180 ug·h / mL, about 160 to about 180 ug·h / mL, or about 170 to about 180 ug·h / mL.

[0158] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 170 ug·h / mL, about 140 to about 170 ug·h / mL, about 150 to about 170 ug·h / mL, or about 160 to about 170 ug·h / mL.

[0159] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 130 to about 160 ug·h / mL, about 140 to about 160 ug·h / mL, or about 150 to about 160 ug·h / mL.

[0160] In some embodiments, the antisense oligomeric CPP conjugate, or a pharma- ceutically acceptable salt thereof, is administered at a dose effective to provide a mean AUC that is about 130 to about 150 ug·h / mL, or about 140 to about 150 ug·h / mL.

[0161] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 140 to about 240 ug·h / mL, about 140 to about 230 ug·h / mL, about 140 to about 220 ug·h / mL, about 140 to about 210 ug·h / mL, about 140 to about 200 ug·h / mL, about 140 to about 190 ug·h / mL, about 140 to about 180 ug·h / mL, about 140 to about 170 ug·h / mL, about 140 to about 160 ug·h / mL, or about 140 to about 150 ug·h / mL.

[0162] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 150 to about 240 ug·h / mL, about 150 to about 230 ug·h / mL, about 150 to about 220 ug·h / mL, about 150 to about 210 ug·h / mL, about 150 to about 200 ug·h / mL, about 150 to about 190 ug·h / mL, about 150 to about 180 ug·h / mL, about 150 to about 170 ug·h / mL, or about 150 to about 160 ug·h / mL.

[0163] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 160 to about 240 ug·h / mL, about 160 to about 230 ug·h / mL, about 160 to about 220 ug·h / mL, about 160 to about 210 ug·h / mL, about 160 to about 200 ug·h / mL, about 160 to about 200 ug·h / mL, about 160 to about 180 ug·h / mL, or about 160 to about 170 ug·h / mL.

[0164] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 170 to about 240 ug·h / mL, about 170 to about 230 ug·h / mL, about 170 to about 220 ug·h / mL, about 170 to about 210 ug·h / mL, about 170 to about 200 ug·h / mL, about 170 to about 190 ug·h / mL, or about 170 to about 180 ug·h / mL.

[0165] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 180 to about 240 ug·h / mL, about 180 to about 230 ug·h / mL, about 180 to about 220 ug·h / mL, or about 180 to about 210 ug·h / mL, about 180 to about 200 ug·h / mL, or about 180 to about 190 ug·h / mL.

[0166] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 190 to about 240 ug·h / mL, about 190 to about 230 ug·h / mL, about 190 to about 220 ug·h / mL, about 190 to about 210 ug·h / mL, or about 190 to about 200 ug·h / mL.

[0167] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 200 to about 240 ug·h / mL, about 200 to about 230 ug·h / mL, about 200 to about 220 ug·h / mL, or about 200 to about 210 ug·h / mL.

[0168] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC that is about 210 to about 240 ug·h / mL, about 210 to about 230 ug·h / mL, or about 210 to about 220 ug·h / mL.

[0169] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 200 to about 500 ug·h / mL, about 200 to about 490 ug·h / mL, about 200 to about 480 ug·h / mL, about 200 to about 470 ug·h / mL, about 200 to about 460 ug·h / mL, about 200 to about 450 ug·h / mL, about 200 to about 440 ug·h / mL, about 200 to about 430 ug·h / mL, about 200 to about 420 ug·h / mL, about 200 to about 410 ug·h / mL, about 200 to about 400 ug·h / mL, about 200 to about 390 ug·h / mL, about 200 to about 380 ug·h / mL, about 200 to about 370 ug·h / mL, about 200 to about 380 ug·h / mL, about 200 to about 390 ug·h / mL, about 200 to about 480 ug·h / mL, about 200 to about 490 ug·h / mL, about 200 to about 500 ug·h / mL, about 200 to about 510 ug·h / mL, about 200 to about 520 ug·h / mL, about 200 to about 530 ug·h / mL, about 200 to about 540 ug·h / mL, about 200 to about 550 ug·h / mL, about 200 to about 560 ug·h / mL, about 200 to about 570 ug·h / mL, about 200 to about 60ug·h / mL, approximately 200 to approximately 350ug·h / mL, approximately 200 to approximately 340ug·h / mL, approximately 200 to approximately 330ug·h / mL, approximately 200 to approximately 320ug· h / mL, about 200 to about 310 ug h / mL, about 200 to about 300 ug h / mL, about 200 to about 290 ug h / mL, about 200 to about 280 ug h / mL, The dose is administered at an effective dose to provide a mean AUC that is about 200 to about 270 ug·h / mL, about 200 to about 260 ug·h / mL, about 200 to about 250 ug·h / mL, about 200 to about 240 ug·h / mL, about 200 to about 230 ug·h / mL, about 200 to about 220 ug·h / mL, or about 200 to about 210 ug·h / mL.

[0170] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 210 to about 500 ug·h / mL, about 210 to about 490 ug·h / mL, about 210 to about 480 ug·h / mL, about 210 to about 470 ug·h / mL, about 210 to about 460 ug·h / mL, about 210 to about 450 ug·h / mL, about 210 to about 440 ug·h / mL, about 210 to about 430 ug·h / mL, about 210 to about 420 ug·h / mL, about 210 to about 410 ug·h / mL, about 210 to about 400 ug·h / mL, about 210 to about 390 ug·h / mL, about 210 to about 380 ug·h / mL, or about 210 to about 370 ug·h / mL. , about 210 to about 360 ug·h / mL, about 210 to about 350 ug·h / mL, about 210 to about 340 ug·h / mL, about 210 to about 330 ug·h / mL, about 210 to about 320 ug·h / mL, about 210 to about 310 ug·h / mL, about 210 to about 300 ug·h / mL, about 210 to about 290 ug·h / mL, about 210 to about 280 ug·h / mL, about 210 to about 270 ug·h / mL, about 210 to about 260 ug·h / mL, about 210 to about 250 ug·h / mL, about 210 to about 240 ug·h / mL, about 210 to about 230 ug·h / mL, or about 210 to about 220 ug·h / mL.

[0171] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 220 to about 500 ug·h / mL, about 220 to about 490 ug·h / mL, about 220 to about 480 ug·h / mL, about 220 to about 470 ug·h / mL, about 220 to about 460 ug·h / mL, about 220 to about 450 ug·h / mL, about 220 to about 440 ug·h / mL, about 220 to about 430 ug·h / mL, about 220 to about 420 ug·h / mL, about 220 to about 410 ug·h / mL, about 220 to about 400 ug·h / mL, about 220 to about 390 ug·h / mL, about 220 to about 380 ug·h / mL, about 220 to about 370ug·h / mL, about 220 to about 360ug·h / mL, about 220 to about 350ug·h / mL, about 220 to about 340ug·h / mL, about 220 to about 330ug·h / mL, about 220 to about 320ug·h / mL, about 220 to about 310ug·h / mL, about 220 to about 300ug·h / mL, about 220 to about 290ug·h / mL, about 220 to about 280ug·h / mL, about 220 to about 270ug·h / mL, about 220 to about 260ug·h / mL, about 220 to about 250ug·h / mL, about 220 to about 240ug·h / mL, or about 220 to about 230ug·h / mL.

[0172] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 230 to about 500 ug·h / mL, about 230 to about 490 ug·h / mL, about 230 to about 480 ug·h / mL, about 230 to about 470 ug·h / mL, about 230 to about 460 ug·h / mL, about 230 to about 450 ug·h / mL, about 230 to about 440 ug·h / mL, about 230 to about 430 ug·h / mL, about 230 to about 420 ug·h / mL, about 230 to about 410 ug·h / mL, about 230 to about 400 ug·h / mL, about 230 to about 390 ug·h / mL, about 230 to about 380 ug·h / mL 230 to about 370 ug·h / mL, about 230 to about 360 ug·h / mL, about 230 to about 350 ug·h / mL, about 230 to about 340 ug·h / mL, about 230 to about 330 ug·h / mL, about 230 to about 320 ug·h / mL, about 230 to about 310 ug·h / mL, about 230 to about 300 ug·h / mL, about 230 to about 290 ug·h / mL, about 230 to about 280 ug·h / mL, about 230 to about 270 ug·h / mL, about 230 to about 260 ug·h / mL, about 230 to about 250 ug·h / mL, or about 230 to about 240 ug·h / mL.

[0173] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 240 to about 500 ug·h / mL, about 240 to about 490 ug·h / mL, about 240 to about 480 ug·h / mL, about 240 to about 470 ug·h / mL, about 240 to about 460 ug·h / mL, about 240 to about 450 ug·h / mL, about 240 to about 440 ug·h / mL, about 240 to about 430 ug·h / mL, about 240 to about 420 ug·h / mL, about 240 to about 410 ug·h / mL, about 240 to about 400 ug·h / mL, about 240 to about 390 ug·h / mL, about 240 to about 380ug·h / mL, about 240 to about 370ug·h / mL, about 240 to about 360ug·h / mL, about 240 to about 350ug·h / mL, about 240 to about 340ug·h / mL, about 240 to about 330ug·h / mL, about 240 to about 320ug·h / mL, about 240 to about 310ug·h / mL, about 240 to about 300ug·h / mL, about 240 to about 290ug·h / mL, about 240 to about 280ug·h / mL, about 240 to about 270ug·h / mL, about 240 to about 260ug·h / mL, or about 240 to about 250ug·h / mL.

[0174] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 300 to about 500 ug·h / mL, about 300 to about 490 ug·h / mL, about 300 to about 480 ug·h / mL, about 300 to about 470 ug·h / mL, about 300 to about 460 ug·h / mL, about 300 to about 450 ug·h / mL, about 300 to about 440 ug·h / mL, about 300 to about 430 ug·h / mL, about 300 to about 420 ug·h / mL, or about 300 to about 440 ug·h / mL. 300 to about 350 ug h / mL, about 300 to about 340 ug h / mL, about 300 to about 330 ug h / mL, about 300 to about 320 ug h / mL, or about 300 to about 310 ug h / mL.

[0175] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has a cytotoxicity of about 325 to about 500 ug·h / mL, about 325 to about 490 ug·h / mL, about 325 to about 480 ug·h / mL, about 325 to about 470 ug·h / mL, about 325 to about 460 ug·h / mL, about 325 to about 450 ug·h / mL, about 325 to about 440 ug·h / mL, about 325 to about 430 ug·h / mL, about 325 to about The present invention is administered at a dose effective to provide a mean AUC of about 420 ug·h / mL, about 325 to about 410 ug·h / mL, about 325 to about 400 ug·h / mL, about 325 to about 390 ug·h / mL, about 325 to about 380 ug·h / mL, about 325 to about 370 ug·h / mL, about 325 to about 360 ug·h / mL, about 325 to about 350 ug·h / mL, about 325 to about 340 ug·h / mL, or about 325 to about 330 ug·h / mL.

[0176] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 330 to about 500 ug·h / mL, about 330 to about 490 ug·h / mL, about 330 to about 480 ug·h / mL, about 330 to about 470 ug·h / mL, about 330 to about 460 ug·h / mL, about 330 to about 450 ug·h / mL, about 330 to about 440 ug·h / mL, about 330 to about 430 ug·h / mL L, about 330 to about 420 ug·h / mL, about 330 to about 410 ug·h / mL, about 330 to about 400 ug·h / mL, about 330 to about 390 ug·h / mL, about 330 to about 380 ug·h / mL, about 330 to about 370 ug·h / mL, about 330 to about 360 ug·h / mL, about 330 to about 350 ug·h / mL, or about 330 to about 340 ug·h / mL.

[0177] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has a cytotoxicity of about 335 to about 500 ug·h / mL, about 335 to about 490 ug·h / mL, about 335 to about 480 ug·h / mL, about 335 to about 470 ug·h / mL, about 335 to about 460 ug·h / mL, about 335 to about 450 ug·h / mL, about 335 to about 440 ug·h / mL, about 335 to about 430 ug·h / mL L, about 335 to about 420 ug·h / mL, about 335 to about 410 ug·h / mL, about 335 to about 400 ug·h / mL, about 335 to about 390 ug·h / mL, about 335 to about 380 ug·h / mL, about 335 to about 370 ug·h / mL, about 335 to about 360 ug·h / mL, about 335 to about 350 ug·h / mL, or about 335 to about 340 ug·h / mL.

[0178] In some embodiments, the antisense oligomer CPP conjugate or a pharma- ceutically acceptable salt thereof has a cytotoxicity of about 340 to about 500 ug·h / mL, about 340 to about 490 ug·h / mL, about 340 to about 480 ug·h / mL, about 340 to about 470 ug·h / mL, about 340 to about 460 ug·h / mL, about 340 to about 450 ug·h / mL, about 340 to about 440 ug·h / mL, about 340 to about The therapeutic dose is administered at a dose effective to provide a mean AUC of about 430 ug·h / mL, about 340 to about 420 ug·h / mL, about 340 to about 410 ug·h / mL, about 340 to about 400 ug·h / mL, about 340 to about 390 ug·h / mL, about 340 to about 380 ug·h / mL, about 340 to about 370 ug·h / mL, about 340 to about 360 ug·h / mL, or about 340 to about 350 ug·h / mL.

[0179] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has a cytotoxicity of about 100 ug·h / mL, about 110 ug·h / mL, about 120 ug·h / mL, about 130 ug·h / mL, about 140 ug·h / mL, about 150 ug·h / mL, about 160 ug·h / mL, about 170 ug·h / mL, about 171 ug·h / mL, about 175 ug·h / mL, about 178 ug·h / mL, about 180 ug·h / mL, about 190 ug·h / mL, about 200 ug·h / mL, about 210 ug·h / mL, about 220 ug·h / mL, about 230 ug·h / mL, about 240 ug·h / mL, about 250 ug·h / mL, about 260 ug·h / mL, about 270 ug·h / mL, about 280 ug·h / mL, or about 290 ug·h / mL. h / mL, approximately 290ug·h / mL, approximately 296ug·h / mL, approximately 298ug·h / mL, approximately 300ug·h / mL, approximately 310ug·h / mL, approximately 320ug·h / mL , approximately 330ug·h / mL, approximately 340ug·h / mL, approximately 350ug·h / mL, approximately 360ug·h / mL, approximately 370ug·h / mL, approximately 380ug·h / mL, approximately 390 The subject is administered at a dose effective to provide a mean AUC of about 400 ug·h / mL, about 410 ug·h / mL, about 420 ug·h / mL, about 430 ug·h / mL, about 450 ug·h / mL, about 460 ug·h / mL, about 470 ug·h / mL, about 480 ug·h / mL, about 490 ug·h / mL, or about 500 ug·h / mL.

[0180] In some embodiments, the antisense oligomeric CPP conjugate or a pharma- ceutical acceptable salt thereof is administered to a patient at a target mean Cmax of the conjugate or a pharma- ceutical acceptable salt thereof that maximizes dystrophin expression and minimizes hypomagnesemia in the patient. max The therapeutic agent is administered in an amount effective to provide

[0181] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytosolic concentration of about 70 to about 150 ug / ml, about 70 to about 140 ug / ml, about 70 to about 130 ug / ml, about 70 to about 120 ug / ml, about 70 to about 110 ug / ml, about 70 to about 100 ug / ml, about 70 to about 90 ug / ml, or about 70 to about 80 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0182] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytosolic concentration of about 75 to about 150 ug / ml, about 75 to about 140 ug / ml, about 75 to about 130 ug / ml, about 75 to about 120 ug / ml, about 75 to about 110 ug / ml, about 75 to about 100 ug / ml, about 75 to about 90 ug / ml, or about 75 to about 80 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0183] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytotoxicity of about 80 to about 150 ug / ml, about 80 to about 140 ug / ml, about 80 to about 130 ug / ml, about 80 to about 120 ug / ml, about 80 to about 110 ug / ml, about 80 to about 100 ug / ml, or about 80 to about 90 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0184] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytotoxicity of about 85 to about 150 ug / ml, about 85 to about 140 ug / ml, about 85 to about 130 ug / ml, about 85 to about 120 ug / ml, about 85 to about 110 ug / ml, about 85 to about 100 ug / ml, or about 85 to about 90 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0185] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytotoxicity of about 90 to about 150 ug / ml, about 90 to about 140 ug / ml, about 90 to about 130 ug / ml, about 90 to about 120 ug / ml, about 90 to about 110 ug / ml, or about 90 to about 100 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0186] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytotoxicity of about 95 to about 150 ug / ml, about 95 to about 140 ug / ml, about 95 to about 130 ug / ml, about 95 to about 120 ug / ml, about 95 to about 110 ug / ml, or about 95 to about 100 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0187] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytotoxicity of about 100 to about 150 ug / ml, about 100 to about 140 ug / ml, about 100 to about 130 ug / ml, about 100 to about 120 ug / ml, or about 100 to about 110 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0188] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytosolic concentration of about 130 to about 250 ug / ml, about 140 to about 250 ug / ml, about 150 to about 250 ug / ml, about 160 to about 250 ug / ml, about 170 to about 250 ug / ml, about 180 to about 250 ug / ml, about 190 to about 250 ug / ml, about 200 to about 250 ug / ml, about 210 to about 250 ug / ml, about 220 to about 250 ug / ml, about 230 to about 250 ug / ml, or about 240 to about 250 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0189] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average cytosolic concentration of about 130 to about 200 ug / ml, about 140 to about 200 ug / ml, about 150 to about 200 ug / ml, about 160 to about 200 ug / ml, about 170 to about 200 ug / ml, about 180 to about 200 ug / ml, or about 190 to about 200 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0190] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average IgG of about 130 to about 190 ug / ml, about 140 to about 190 ug / ml, about 150 to about 190 ug / ml, about 160 to about 190 ug / ml, about 170 to about 190 ug / ml, or about 180 to about 190 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0191] In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof has an average IgG of about 130 to about 180 ug / ml, about 140 to about 180 ug / ml, about 150 to about 180 ug / ml, about 160 to about 180 ug / ml, or about 170 to about 180 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax In some embodiments, the antisense oligomer CPP conjugate or pharma- ceutically acceptable salt thereof is administered in a dose effective to provide an average IgG antibody concentration of about 130 to about 170 ug / ml, about 140 to about 170 ug / ml, about 150 to about 170 ug / ml, or about 160 to about 170 ug / ml of the conjugate or pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0192] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is at a concentration of about 80ug / ml, about 85ug / ml, about 90ug / ml, about 95ug / ml, about 100ug / ml, about 105ug / ml, about 110ug / ml, about 115ug / ml, about 120ug / ml, about 125ug / ml, about 130ug / ml, about 135ug / ml, about 140ug / ml, about 145ug / ml, about 150ug / ml, about 155ug / ml, about 160ug / ml 1, about 165 ug / ml, about 170 ug / ml, about 175 ug / ml, about 180 ug / ml, about 185 ug / ml, about 190 ug / ml, about 195 ug / ml, about 200 ug / ml, about 205 ug / ml, about 210 ug / ml, about 215 ug / ml, about 220 ug / ml, about 225 ug / ml, about 230 ug / ml, about 235 ug / ml, about 240 ug / ml, about 245 ug / ml, or about 250 ug / ml of the conjugate or a pharma- ceutically acceptable salt thereof. Cmax The therapeutic agent is administered in an amount effective to provide

[0193] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose of about 300 mg to about 900 mg, about 300 mg to about 850 mg, about 300 mg to about 800 mg, about 300 mg to about 850 mg, about 300 mg to about 800 mg, about 300 mg to about 750 mg, about 300 mg to about 700 mg, about 300 mg to about 650 mg, about 300 mg to about 600 mg, about 300 mg to about 550 mg, about 300 mg to about 500 mg, about 300 mg to about 450 mg, about 300 mg to about 400 mg, or about 300 mg.

[0194] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg to about 900 mg, about 300 mg to about 850 mg, about 300 mg to about 800 mg, about 300 mg to about 850 mg, about 300 mg to about 800 mg, about 300 mg to about 750 mg, about 300 mg to about 700 mg, about 300 mg to about 650 mg, about 300 mg to about 600 mg, about 300 mg to about 550 mg, about 300 mg to about 500 mg, about 300 mg to about 450 mg, about 300 mg to about 400 mg, or about 300 mg to about 350 mg of the .6HCl salt of the conjugate.

[0195] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 350 mg to about 900 mg, about 400 mg to about 900 mg, about 500 mg to about 900 mg, about 550 mg to about 900 mg, about 600 mg to about 800 mg, about 650 mg to about 900 mg, about 700 mg to about 900 mg, about 750 mg to about 900 mg, about 800 mg to about 900 mg, or about 850 mg to about 900 mg of the .6HCl salt of the conjugate.

[0196] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 350 mg to about 850 mg, about 400 mg to about 850 mg, about 500 mg to about 850 mg, about 550 mg to about 850 mg, about 600 mg to about 800 mg, about 650 mg to about 850 mg, about 700 mg to about 850 mg, about 750 mg to about 850 mg, or about 800 mg to about 850 mg of the .6HCl salt of the conjugate.

[0197] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 350 mg to about 800 mg, about 400 mg to about 800 mg, about 500 mg to about 800 mg, about 550 mg to about 800 mg, about 600 mg to about 800 mg, about 650 mg to about 800 mg, about 700 mg to about 800 mg, or about 750 mg to about 800 mg of the .6HCl salt of the conjugate.

[0198] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 350 mg to about 750 mg, about 400 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg to about 750 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, or about 700 mg to about 750 mg of the .6HCl salt of the conjugate.

[0199] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 350 mg to about 700 mg, about 400 mg to about 700 mg, about 500 mg to about 700 mg, about 550 mg to about 700 mg, about 600 mg to about 700 mg, or about 650 mg to about 700 mg of the .6HCl salt of the conjugate.

[0200] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 400 mg to about 900 mg, about 400 mg to about 850 mg, about 400 mg to about 800 mg, about 400 mg to about 750 mg, about 400 mg to about 700 mg, about 400 mg to about 650 mg, about 400 mg to about 600 mg, about 400 mg to about 550 mg, about 400 mg to about 500 mg, or about 400 mg to about 450 mg of the .6HCl salt of the conjugate.

[0201] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1500 mg, about 900 mg to about 1500 mg, about 850 mg to about 1500 mg, or about 800 mg to about 1500 mg of the .6HCl salt of the conjugate.

[0202] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1450 mg, about 900 mg to about 1450 mg, about 850 mg to about 1450 mg, or about 800 mg to about 1450 mg of the .6HCl salt of the conjugate.

[0203] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1400 mg, about 900 mg to about 1400 mg, about 850 mg to about 1400 mg, or about 800 mg to about 1400 mg of the .6HCl salt of the conjugate.

[0204] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1350 mg, about 900 mg to about 1350 mg, about 850 mg to about 1350 mg, or about 800 mg to about 1350 mg of the .6HCl salt of the conjugate.

[0205] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1300 mg, about 900 mg to about 1300 mg, about 850 mg to about 1300 mg, or about 800 mg to about 1300 mg of the .6HCl salt of the conjugate.

[0206] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg to about 1250 mg, about 900 mg to about 1250 mg, about 850 mg to about 1250 mg, or about 800 mg to about 1250 mg of the .6HCl salt of the conjugate.

[0207] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 950 mg to about 1200 mg, about 900 mg to about 1200 mg, about 850 mg to about 1200 mg, about 800 mg to about 1200 mg, about 900 mg to about 1100 mg, about 900 mg to about 1000 mg, about 800 mg to about 1100 mg, or about 800 mg to about 1000 mg of the .6HCl salt of the conjugate.

[0208] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1000 mg to about 1500 mg, about 1000 mg to about 1450 mg, about 1000 mg to about 1400 mg, about 1000 mg to about 1350 mg, about 1000 mg to about 1300 mg, about 1000 mg to about 1250 mg, about 1000 mg to about 1200 mg, about 1000 mg to about 1150 mg, or about 1000 mg to about 1100 mg of the .6HCl salt of the conjugate.

[0209] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the .6HCl salt of the conjugate.

[0210] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the .6HCl salt of the conjugate.

[0211] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered according to the patient's weight. The following weight ranges are contemplated by the present disclosure: about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, about 18 kg to about 50 kg, about 25 kg to about 50 kg, about 30 kg to about 50 kg, about 35 kg to about 50 kg, about 40 kg to about 50 kg, about 50 kg to about 100 kg, about 55 kg to about 100 kg, about 60 kg to about 100 kg, about 65 kg g to about 100 kg, about 70 kg to about 100 kg, about 75 kg to about 100 kg, about 80 kg to about 100 kg, about 85 kg to about 100 kg, about 90 kg to about 100 kg, about 50 kg to about 90 kg, about 55 kg to about 90 kg, about 60 kg to about 90 kg, about 65 kg to about 90 kg, about 70 kg to about 90 kg, about 75 kg to about 90 kg, about 80 kg to about 90 kg, or about 85 kg to about 90 kg.

[0212] The following weight ranges are also contemplated by this disclosure: 5 kg to less than 25 kg, 10 kg to less than 25 kg, 15 kg to less than 25 kg, 20 kg to less than 25 kg, 18 kg to less than 50 kg, 25 kg to less than 50 kg, 30 kg to less than 50 kg, 35 kg to less than 50 kg, 40 kg to less than 50 kg, 50 kg, 50 kg to less than 100 kg, 55 kg to less than 100 kg, 60 kg to less than 100 kg, 65 kg g or more but less than 100kg, 70kg or more but less than 100kg, 75kg or more but less than 100kg, 80kg or more but less than 100kg, 85kg or more but less than 100kg, 90kg or more but less than 100kg, 50kg or more but less than 90kg, 55kg or more but less than 90kg, 60kg or more but less than 90kg, 65kg or more but less than 90kg, 70kg or more but less than 90kg, 75kg or more but less than 90kg, 80kg or more but less than 90kg, or 85kg or more but less than 90kg.

[0213] The following weight ranges are also contemplated by the present disclosure: about 5 kg to about 20 kg, about 10 kg to about 20 kg, about 15 kg to about 20 kg, about 25 kg to about 40 kg, about 30 kg to about 40 kg, about 35 kg to about 40 kg, about 18 kg to about 50 kg, about 50 kg to about 90 kg, about 55 kg to about 90 kg, about 60 kg to about 90 kg, about 65 kg to about 90 kg, about 70 kg to about 90 kg, about 75 kg to about 90 kg, about 80 kg to about 90 kg, or about 85 kg to about 90 kg.

[0214] The following weight ranges are also contemplated by this disclosure: 5 kg to less than 20 kg, 10 kg to less than 20 kg, 15 kg to less than 20 kg, 25 kg to less than 40 kg, 30 kg to less than 40 kg, 35 kg to less than 40 kg, 50 kg to less than 90 kg, 55 kg to less than 90 kg, 60 kg to less than 90 kg, 65 kg to less than 90 kg, 70 kg to less than 90 kg, 75 kg to less than 90 kg, 80 kg to less than 90 kg, or 85 kg to less than 90 kg.

[0215] The following weight ranges are also contemplated by the present disclosure: about 5 kg to about 15 kg, about 10 kg to about 15 kg, about 25 kg to about 30 kg, about 30 kg to about 35 kg, about 30 kg to about 45 kg, about 50 kg to about 85 kg, about 55 kg to about 85 kg, about 60 kg to about 85 kg, about 65 kg to about 85 kg, about 70 kg to about 85 kg, about 75 kg to about 85 kg, about 50 kg to about 75 kg, about 55 kg to about 75 kg, about 60 kg to about 75 kg, about 65 kg to about 75 kg, about 70 kg to about 75 kg, or about 80 kg to about 85 kg.

[0216] The following weight ranges are also contemplated by this disclosure: 5 kg to less than 15 kg, 10 kg to less than 15 kg, 25 kg to less than 30 kg, 30 kg to less than 35 kg, 30 kg to less than 45 kg, 50 kg to less than 85 kg, 55 kg to less than 85 kg, 60 kg to less than 85 kg, 65 kg to less than 85 kg, 70 kg to less than 85 kg, 75 kg to less than 85 kg, 50 kg to less than 75 kg, 55 kg to less than 75 kg, 60 kg to less than 75 kg, 65 kg to less than 75 kg, 70 kg to less than 75 kg, or 80 kg to less than 85 kg.

[0217] The following weight ranges are also contemplated by the present disclosure: about 10 kg to about 50 kg, about 10 kg to about 45 kg, about 10 kg to about 40 kg, about 10 kg to about 35 kg, about 15 kg to about 50 kg, about 15 kg to about 45 kg, about 15 kg to about 40 kg, about 15 kg to about 35 kg, about 20 kg to about 50 kg, about 20 kg to about 45 kg, about 20 kg to about 40 kg, about 20 kg to about 35 kg, about 25 kg to about 50 kg, about 25 kg to about 45 kg, about 25 kg to about 40 kg, or about 25 kg to about 35 kg.

[0218] The following weight ranges are also contemplated by this disclosure: 10 kg to less than 50 kg, 10 kg to less than 45 kg, 10 kg to less than 40 kg, 10 kg to less than 35 kg, 15 kg to less than 50 kg, 15 kg to less than 45 kg, 15 kg to less than 40 kg, 15 kg to less than 35 kg, 20 kg to less than 50 kg, 20 kg to less than 45 kg, 20 kg to less than 40 kg, 20 kg to less than 35 kg, 25 kg to less than 50 kg, 25 kg to less than 45 kg, 25 kg to less than 40 kg, or 25 kg to less than 35 kg.

[0219] The following weight ranges are also contemplated by the present disclosure: about 20 kg to about 110 kg, about 30 kg to about 110 kg, about 40 kg to about 110 kg, about 50 kg to about 110 kg, about 60 kg to about 110 kg, about 20 kg to about 100 kg, about 30 kg to about 100 kg, about 40 kg to about 100 kg, about 50 kg to about 100 kg, about 60 kg to about 100 kg, about 20 kg to about 90 kg, about 30 kg to about 90 kg, about 40 kg to about 90 kg, about 50 kg to about 90 kg, or about 60 kg to about 90 kg.

[0220] The following weight ranges are also contemplated by this disclosure: 20 kg to less than 110 kg, 30 kg to less than 110 kg, 40 kg to less than 110 kg, 50 kg to less than 110 kg, 60 kg to less than 110 kg, 20 kg to less than 100 kg, 30 kg to less than 100 kg, 40 kg to less than 100 kg, 50 kg to less than 100 kg, 60 kg to less than 100 kg, 20 kg to less than 90 kg, 30 kg to less than 90 kg, 40 kg to less than 90 kg, 50 kg to less than 90 kg, or 60 kg to less than 90 kg.

[0221] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered according to weight ranges at a dose effective to achieve a mean AUC of about 100 to about 200 ug·h / mL in patients within a particular weight range. In other embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered according to weight ranges at a dose effective to achieve a mean AUC of about 120 to about 240 ug·h / mL in patients within a particular weight range. In other embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered according to weight ranges at a dose effective to achieve a mean AUC of about 200 to about 500 ug·h / mL in patients within a particular weight range.

[0222] In some embodiments, the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, is administered according to the following schedule: vii) about 300 mg to about 700 mg or about 900 mg to about 1200 mg once every 4 weeks for a patient weighing about 10 kg to about 25 kg; viii) about 600 mg to about 800 mg or about 1000 mg to about 1300 mg once every 4 weeks for a patient weighing about 25 kg to about 50 kg, or ix) about 700 mg to about 900 mg or about 1200 mg to about 1500 mg once every 4 weeks for a patient weighing about 50 kg to about 100 kg; to achieve a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0223] In some embodiments, the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, is administered according to the following schedule: x) about 500 mg to about 700 mg or about 1000 mg to about 1200 mg once every 4 weeks for a patient weighing about 15 kg to about 40 kg, or xi) about 600 mg to about 800 mg or about 1100 mg to about 1400 mg once every 4 weeks for a patient weighing about 40 kg to about 100 kg; to achieve a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0224] In some embodiments, the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, is administered according to the following schedule: xii) about 750 mg to about 1,250 mg once every 4 weeks for patients weighing about 18 kg or more and less than about 50 kg, or xiii) About 850 mg to about 1,350 mg once every 4 weeks for patients weighing about 50 kg or more; according to the above, administered at a dose equivalent to that of the 6HCl salt of the conjugate to achieve a mean AUC of about 120 to about 240 ug·h / mL or about 200 to about 500 ug·h / mL.

[0225] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a concentration of about 300 mg to about 750 mg, about 350 mg to about 750 mg, about 400 mg to about 750 mg, about 450 mg to about 750 mg, about 400 mg to about 750 mg, about 450 mg to about 750 mg, about 500 mg to about 750 mg, about 550 mg to about 750 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, 300 mg to about 700 mg, about 350 mg to about 700 mg, about 400 mg to about 700 mg, about 450 mg to about 700 mg, about 400 mg to about 700 mg, about 450 mg to about 700 mg, about 500 mg to about 750 mg, The .6HCl salt of the conjugate is administered at a dose equivalent to about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, 5 kg to about 20 kg, about 10 kg to about 20 kg, or about 15 kg to about 20 kg in a dose equivalent to about 5 kg to about 700 mg, about 550 mg to about 700 mg, about 600 mg to about 700 mg, about 650 mg to about 700 mg, about 300 mg to about 650 mg, about 350 mg to about 650 mg, about 400 mg to about 650 mg, about 450 mg to about 650 mg, about 500 mg to about 650 mg, about 550 mg to about 650 mg, or about 600 mg to about 650 mg of the .6HCl salt of the conjugate to a patient weighing about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, 5 kg to about 20 kg, about 10 kg to about 20 kg, or about 15 kg to about 20 kg.

[0226] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 600 mg to about 800 mg, about 650 mg to about 800 mg, about 700 mg to about 800 mg, about 750 mg to about 800 mg, about 600 mg to about 750 mg, about 650 mg to about 750 mg, about 700 mg to about 750 mg, 600 mg to about 700 mg, or about 650 mg to about 700 mg of the .6HCl salt of the conjugate, The present invention is directed to a patient weighing about 25kg to about 40kg, about 30kg to about 50kg, about 35kg to about 50kg, about 40kg to about 50kg, about 25kg to about 40kg, about 30kg to about 40kg, about 35kg to about 40kg, about 25kg to about 30kg, about 30kg to about 45kg, about 35kg to about 45kg, about 40kg to about 45kg, about 30kg to about 35kg, about 35kg to about 45kg, about 25kg to about 45kg, about 25kg to about 35kg, or about 35kg to about 45kg.

[0227] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered to a patient weighing greater than about 18 kg and less than about 50 kg at a dose equivalent to about 750 mg to about 1250 mg of the .6HCl salt of the conjugate. In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered to a patient weighing greater than about 50 kg at a dose equivalent to about 850 mg to about 1350 mg of the .6HCl salt of the conjugate.

[0228] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 700 mg to about 900 mg, about 750 mg to about 900 mg, about 800 mg to about 900 mg, or about 850 mg to about 900 mg, about 700 mg to about 850 mg, about 750 mg to about 850 mg, or about 800 mg to about 850 mg, about 700 mg to about 750 mg, about 750 mg to about 800 mg, about 750 mg to about 900 mg, or about 800 mg to about 900 mg of the .6HCl salt of the conjugate, It is administered to a patient weighing about 50kg to about 100kg, about 55kg to about 100kg, about 60kg to about 100kg, about 65kg to about 100kg, about 70kg to about 100kg, about 75kg to about 100kg, about 80kg to about 100kg, about 85kg to about 100kg, or about 90kg to about 100kg, about 50kg to about 90kg, about 55kg to about 90kg, about 60kg to about 90kg, about 65kg to about 90kg, about 70kg to about 90kg, about 75kg to about 90kg, about 80kg to about 90kg, or about 85kg to about 90kg.

[0229] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 900 mg to about 1200 mg, about 850 mg to about 1200 mg, about 800 mg to about 1200 mg, about 900 mg to about 1100 mg, about 900 mg to about 1000 mg, about 800 mg to about 1100 mg, or about 800 mg to about 1000 mg of the .6HCl salt of the conjugate to a patient weighing about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, 5 kg to about 20 kg, about 10 kg to about 20 kg, or about 15 kg to about 20 kg.

[0230] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is about 1000 mg to about 1300 mg, about 1000 mg to about 1250 mg, about 1000 mg to about 1200 mg, about 1000 mg to about 1150 mg, about 1050 mg to about 1100 mg, about 1050 mg to about 1300 mg, about 1050 mg to about 1250 mg, about 1050 mg to about 1200 mg, about 1050 mg to about 1150 mg, about 1050 mg to about 1100 mg, about 1050 mg to about 1250, about 1100 mg to about 1250, about 1150 mg to about 1250, about 1200 mg about 25 kg to about 50 kg, about 30 kg to about 50 kg, about 35 kg to about 50 kg, about 40 kg to about 50 kg, about 25 kg to about 40 kg, about 30 kg to about 40 kg, about 35 kg to about 40 kg, about 25 kg to about 30 kg, about 30 kg to about 45 kg, about 35 kg to about 45 kg, about 40 kg to about 45 kg, about 30 kg to about 35 kg, about 35 kg to about 45 kg, about 25 kg to about 45 kg, about 25 kg to about 35 kg, or about 35 kg to about 45 kg.

[0231] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is about 1200 mg to about 1500 mg, 1000 mg to about 1500 mg, about 1000 mg to about 1450 mg, about 1000 mg to about 1400 mg, about 1000 mg to about 1350 mg, about 1000 mg to about 1300 mg, about 1000 mg to about 1250 mg, about 1200 mg to about 1450 mg, about 1200 mg to about 1400 mg, about 1200 mg to about 1350 mg, about 1200 mg to about 1300 mg, or about 1200 mg to about 1250 mg of the conjugate. It is administered in a dose equivalent to that of the .6HCl salt to a patient weighing about 50kg to about 100kg, about 55kg to about 100kg, about 60kg to about 100kg, about 65kg to about 100kg, about 70kg to about 100kg, about 75kg to about 100kg, about 80kg to about 100kg, about 85kg to about 100kg, or about 90kg to about 100kg, about 50kg to about 90kg, about 55kg to about 90kg, about 60kg to about 90kg, about 65kg to about 90kg, about 70kg to about 90kg, about 75kg to about 90kg, about 80kg to about 90kg, or about 85kg to about 90kg.

[0232] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 550 mg, 600 mg, about 650 mg, or about 700 mg of the .6HCl salt of the conjugate to a patient weighing about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, 5 kg to about 20 kg, about 10 kg to about 20 kg, or about 15 kg to about 20 kg.

[0233] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1050 mg, 1100 mg, 1150 mg, or about 1200 mg of the .6HCl salt of the conjugate to a patient weighing about 5 kg to about 25 kg, about 10 kg to about 25 kg, about 15 kg to about 25 kg, about 20 kg to about 25 kg, 5 kg to about 20 kg, about 10 kg to about 20 kg, or about 15 kg to about 20 kg.

[0234] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 600 mg, about 650 mg, about 700 mg, or about 700 mg of the .6HCl salt of the conjugate to a patient weighing about 25 kg to about 50 kg, about 30 kg to about 50 kg, about 35 kg to about 50 kg, about 40 kg to about 50 kg, about 25 kg to about 40 kg, about 30 kg to about 40 kg, about 35 kg to about 40 kg, about 25 kg to about 30 kg, about 30 kg to about 45 kg, about 35 kg to about 45 kg, about 40 kg to about 45 kg, about 30 kg to about 35 kg, about 35 kg to about 45 kg, about 25 kg to about 45 kg, about 25 kg to about 35 kg, or about 35 kg to about 45 kg.

[0235] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, or about 1300 mg of the .6HCl salt of the conjugate to a patient weighing about 25 kg to about 50 kg, about 30 kg to about 50 kg, about 35 kg to about 50 kg, about 40 kg to about 50 kg, about 25 kg to about 40 kg, about 30 kg to about 40 kg, about 35 kg to about 40 kg, about 25 kg to about 30 kg, about 30 kg to about 45 kg, about 35 kg to about 45 kg, about 40 kg to about 45 kg, about 30 kg to about 35 kg, about 35 kg to about 45 kg, about 25 kg to about 45 kg, about 25 kg to about 35 kg, or about 35 kg to about 45 kg.

[0236] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 700 mg, about 750 mg, about 800 mg, or about 850 mg of the .6HCl salt of the conjugate, about 50 kg to about 100 kg, about 55 kg to about 100 kg, about 60 kg to about 100 kg, about 65 kg to about 100 kg, about 70 kg to about 100 kg, It is administered to a patient weighing about 75kg to about 100kg, about 80kg to about 100kg, about 85kg to about 100kg, or about 90kg to about 100kg, about 50kg to about 90kg, about 55kg to about 90kg, about 60kg to about 90kg, about 65kg to about 90kg, about 70kg to about 90kg, about 75kg to about 90kg, about 80kg to about 90kg, or about 85kg to about 90kg.

[0237] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, or about 1550 mg of the .6HCl salt of the conjugate, about 50 kg to about 100 kg, about 55 kg to about 100 kg, about 60 kg to about 100 kg, about 65 kg to about 100 kg, about 70 kg to about 100 kg, about 75 kg to about 100 kg, about 80 kg to about 100 kg, about 85 kg to about 100 kg, about 90 kg to about 100 kg, about 95 kg to about 100 kg, about 100 kg to about 100 kg, about 110 kg to about 115 kg, about 120 kg to about 125 kg, about 130 kg to about 1350 mg, about 1400 mg, about 1450 mg, about 1500 mg, or about 1550 mg of the .6HCl salt of the conjugate. The present invention is directed to a patient weighing about 100 kg to about 100 kg, about 75 kg to about 100 kg, about 80 kg to about 100 kg, about 85 kg to about 100 kg, or about 90 kg to about 100 kg, about 50 kg to about 90 kg, about 55 kg to about 90 kg, about 60 kg to about 90 kg, about 65 kg to about 90 kg, about 70 kg to about 90 kg, about 75 kg to about 90 kg, about 80 kg to about 90 kg, or about 85 kg to about 90 kg.

[0238] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, or about 1100 mg of the .6HCl salt of the conjugate to a patient weighing about 80 kg to about 100 kg, about 85 kg to about 100 kg, about 90 kg to about 100 kg, about 95 kg to about 100 kg, about 80 kg to about 90 kg, about 75 kg to about 90 kg, about 75 kg to about 95 kg, about 75 kg to about 80 kg, or about 80 kg to about 90 kg.

[0239] In some embodiments, the antisense oligomer conjugate or pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, or about 1400 mg of the .6HCl salt of the conjugate to a patient weighing about 80 kg to about 100 kg, about 85 kg to about 100 kg, about 90 kg to about 100 kg, about 95 kg to about 100 kg, about 80 kg to about 90 kg, about 75 kg to about 90 kg, about 75 kg to about 95 kg, about 75 kg to about 80 kg, or about 80 kg to about 90 kg.

[0240] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 700 mg, about 750 mg, about 800 mg, about 850 mg, about 900 mg, about 950 mg, or about 1000 mg of the .6HCl salt of the conjugate to a patient weighing greater than about 45 kg, about 50 kg, about 55 kg, about 60 kg, or about 65 kg.

[0241] In some embodiments, the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, or about 1450 mg of the .6HCl salt of the conjugate to a patient weighing greater than about 45 kg, about 50 kg, about 55 kg, about 60 kg, or about 65 kg.

[0242] If desired, the antisense oligomer conjugate or its pharma- ceutically acceptable salt may be administered as 2, 3, 4, 5, 6 or more partial doses, which are administered separately at appropriate intervals throughout the day, optionally in unit dosage form. In certain circumstances, the dosage is administered once a day. In certain embodiments, the dosage is administered once or more every 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, or once or more every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, or once or more every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 months, as needed to maintain the desired expression of functional dystrophin protein. In certain embodiments, the dosage is administered once or more every two weeks. In some embodiments, the dosage is administered once every two weeks. In various embodiments, the dosage is administered once or more every month. In certain embodiments, the dosage is a once monthly administration.

[0243] Cell-penetrating peptides (CPPs) As described, cell membrane penetrating peptides (CPPs), e.g., arginine-rich peptide transport moieties, are effective in enhancing the penetration of antisense oligomers into cells and have been shown to cause exon skipping in different muscle groups in animal models.

[0244] Exemplary CPPs are provided in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10] [Table 1-11]

[0245] In some embodiments, the antisense oligonucleotide comprises a substituent "Z" defined as a combination of a CPP and a linker. The linker bridges the CPP at its carboxy terminus to the 3' and / or 5' terminus of the oligonucleotide. In various embodiments, the antisense oligonucleotide may contain only one CPP linked to the 3' terminus of the oligomer. In other embodiments, the antisense oligonucleotide may contain only one CPP linked to the 5' terminus of the oligomer.

[0246] The linker in Z can include, for example, 1, 2, 3, 4, or 5 amino acids. For example, the linker can include 1, 2, 3, 4, or 5 of glycine moieties, and the CPP is attached to the glycine moiety by an amide bond at the CPP carboxy terminus. In one embodiment, the linker includes one glycine moiety.

[0247] In certain embodiments, Z is -C(O)(CH2)5NH-CPP, -C(O)(CH2)2NH-CPP, -C(O)(CH2)2NHC(O)(CH2)5NH-CPP, -C(O)CHNH-CPP, and [ka] wherein the CPP is attached to the linker moiety via an amide bond at the CPP carboxy terminus.

[0248] In various embodiments, the CPP is an arginine-rich peptide, as defined above and shown in Table 1. In certain embodiments, the arginine-rich CPP is -R a (i.e., six arginine residues), where R a is selected from H, acyl, acetyl, benzoyl, and stearoyl. In certain embodiments, R a is acetyl.

[0249] In certain embodiments, Z is —C(O)CHNH—R—R covalently attached to the antisense oligomer of the disclosure at the 5′ and / or 3′ end of the oligomer. a where R a is H, acyl, acetyl, benzoyl, or stearoyl to cap the amino terminus of R6. a is acetyl. In these non-limiting examples, the CPP is -R-R a and the linker is -C(O)CH2NH- (i.e., GLY). Z = -C(O)CH2NH-R6-R a This particular example is also exemplified by the following structure: [ka] In the formula, R a is selected from H, acyl, acetyl, benzoyl, and stearoyl. a is H or acyl. In some embodiments, R a is acetyl.

[0250] In various embodiments, the CPP is represented by the formula -R-R a and [ka] In the formula, R a is selected from H, acyl, acetyl, benzoyl, and stearoyl. In certain embodiments, the CPP is SEQ ID NO: 128. In some embodiments, R a is H or acyl. In some embodiments, R a is acetyl.

[0251] In some embodiments, the CPP is also exemplified by the formula -(RXR)-R a It is. [ka]

[0252] In various embodiments, the CPP is also exemplified by the formula: -R-(FFR)-R a It is. [ka]

[0253] In various embodiments, Z is -C(O)(CH2)5NH-CPP, -C(O)(CH2)2NH-CPP, -C(O)(CH2)2NHC(O)(CH2)5NH-CPP, -C(O)CHNH-CPP, and [ka] wherein the CPP is attached to a linker moiety by an amide bond at the CPP carboxy terminus, and the CPP is [ka] (-R-(FFR)3-R a ), [ka] (-(RXR)4-Ra ), [ka] and (-R6-R a In some embodiments, R a is H or acyl. In some embodiments, R a is acetyl.

[0254] In some embodiments, "-G-R6" or "-G-R6-Ac" is conjugated to the 3' end of an antisense oligomer of the disclosure and has the formula: [ka]

[0255] Antisense oligomeric CPP conjugates In various aspects, the present disclosure provides an antisense oligomer according to formula (I): [ka] or a pharma- ceutically acceptable salt thereof, wherein: n is 1 to 40; each Nu is a nucleobase that, together, forms a targeting sequence that is complementary to an exon annealing site in the dystrophin pre-mRNA; T' is [ka] is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen, R 1 is C1-C6 alkyl.

[0256] In various embodiments, the antisense oligomer conjugates of the present disclosure have formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein: each Nu is a nucleobase that, taken together, forms a targeting sequence, T is [ka] is selected from R 1 is C1-C6 alkyl, The targeting sequence is complementary to an annealing site in the dystrophin pre-RNA.

[0257] In some embodiments, the antisense oligonucleotide conjugate in the composition comprises a sequence complementary to 15-35 nucleobases of a target region of exon 44, exon 45, exon 50, exon 51, exon 52, or exon 53 of the dystrophin pre-mRNA. Oligonucleotide sequences designed to target and skip these dystrophin exons have been described in the art. See, for example, the following published PCT applications and issued U.S. patents: WO2018 / 129384, WO2019 / 060775, WO2020 / 219820, WO2018 / 007475, WO2018 / 091544, WO2020 / 089325, WO2004 / 048570, WO2020 / 028832, WO2017 / 0628 62, U.S. Patent No. 10,683,322, U.S. Patent No. 8,969,551, U.S. Patent No. 10,781,448, U.S. Patent No. 9,988,629, U.S. Patent No. 9,840,706, U.S. Patent No. 10,851,373, WO2020 / 004675, and WO2020 / 0158792 (the disclosures of which sequences are incorporated herein).

[0258] A number of exemplary targeting sequences are described below. These sequences are provided as morpholino targeting sequences and can be incorporated into the antisense oligonucleotide conjugates of Formula I.

[0259] In some embodiments, the targeting sequence is complementary to an exon 51 annealing site in a dystrophin pre-mRNA. In some embodiments, the site is designated as H51A(+66+95). In some embodiments, the targeting sequence is complementary to an exon 45 annealing site in a dystrophin pre-mRNA. In some embodiments, the site is designated as H45A(-03+19). In some embodiments, the targeting sequence is complementary to an exon 53 annealing site in a dystrophin pre-mRNA. In some embodiments, the site is designated as H53A(+36+60).

[0260] In various embodiments, T is [ka] It is.

[0261] In various embodiments, R 1 is CF3, CCl3, CFCl2, CF2Cl, ethyl, CH2CF3, CF2CF3, propyl, isopropyl, butyl, isobutyl, sec-butyl, t-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, 3-methylpentyl, 2,2-dimethylbutyl, or 2,3-dimethylbutyl.

[0262] In some embodiments, the antisense oligomer conjugate of formula (I) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0263] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I).

[0264] In some embodiments, the targeting sequence is 5'-CTCCAACATCAAGGAAGATGGCATTTCTAG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO:31).

[0265] In various embodiments, T is [ka] and the targeting sequence is 5'-CTCCAACATCAAGGAAGATGGCATTTCTAG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO:31).

[0266] In various embodiments, T is [ka] and the targeting sequence is 5'-CTCCAACATCAAGGAAGATGGCATTTCTAG-3' (SEQ ID NO: 32).

[0267] In some embodiments, the targeting sequence is 5'-CAATGCCATCCTGGAGTTCCTG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO:28).

[0268] In various embodiments, T is [ka] and the targeting sequence is 5'-CAATGCCATCCTGGAGTTCCTG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO:28).

[0269] In various embodiments, T is [ka] and the targeting sequence is 5'-CAATGCCATCCTGGAGTTCCTG-3' (SEQ ID NO: 27).

[0270] In some embodiments, the targeting sequence is 5'-GTTGCCTCCGGTTCTGAAGGTGTTC-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO: 30).

[0271] In various embodiments, T is [ka] and the targeting sequence is 5'-GTTGCCTCCGGTTCTGAAGGTGTTC-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO: 30).

[0272] In various embodiments, T is [ka] and the targeting sequence is 5'-GTTGCCTCCGGTTCTGAAGGTGTTC-3' (SEQ ID NO: 29).

[0273] For example, in some embodiments, including some embodiments of formula (I), the antisense oligomer conjugate of the present disclosure has formula (III): [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each Nu is a nucleobase that together form a targeting sequence that is complementary to an exon 51 annealing site in the dystrophin pre-mRNA designated as H51A(+66+95).

[0274] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I).

[0275] In various embodiments, each Nu from 1 to 30 and 5' to 3' is: [Table 4] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is independently [ka] It is.

[0276] In some embodiments, the antisense oligomer conjugate of formula (III) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0277] For example, in some embodiments, including some embodiments of formula (III), the antisense oligomer conjugates of the present disclosure are according to formula (IIIA): [ka] wherein each Nu is a nucleobase that, taken together, forms a targeting sequence that is complementary to an exon 51 annealing site in the dystrophin pre-mRNA designated as H51A(+66+95).

[0278] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I). In various embodiments, each Nu from 1 to 30 and 5' to 3' is: [Table 5] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is [ka] It is.

[0279] In some embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (III) and formula (IIIA), the targeting sequence is 5'-CTCCAACATCAAGGAAGATGGCATTTCTAG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO: 31). In various embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (III) and formula (IIIA), the targeting sequence is 5'-CTCCAACATCAAGGAAGATGGCATTTCTAG-3' (SEQ ID NO: 32).

[0280] In some embodiments, including for example, embodiments of the antisense oligomer conjugates of formula (I), the antisense oligomer conjugates of the present disclosure have the formula (IV): [ka] or a pharma- ceutically acceptable salt thereof.

[0281] In some embodiments, the antisense oligomer conjugate of formula (IV) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0282] In some embodiments, including for example, embodiments of the antisense oligomer conjugates of formula (IV), the antisense oligomer conjugates of the present disclosure are according to formula (IVA). [ka]

[0283] In some embodiments of the present disclosure, including some embodiments of the antisense oligomer conjugate of formula (I) and embodiments of the antisense oligomer conjugate of formula (IV), the antisense oligomer conjugate has the formula (IVB): [ka] or a pharma- ceutically acceptable salt thereof.

[0284] In some embodiments, the antisense oligomer conjugate of formula (IVB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0285] In some embodiments, including for example, the embodiment of the antisense oligomer conjugate of formula (IVB), the antisense oligomer conjugate of the present disclosure is according to formula (IVC). [ka]

[0286] For example, in some embodiments, including some embodiments of formula (I), the antisense oligomer conjugate of the present disclosure has formula (V): [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each Nu is a nucleobase that together form a targeting sequence that is complementary to an exon 45 annealing site in the dystrophin pre-mRNA designated as H45A(-03+19).

[0287] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I). In various embodiments, each Nu from 1 to 22 and 5' to 3' is: [Table 6] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is independently [ka] It is.

[0288] In some embodiments, the antisense oligomer conjugate of formula (V) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0289] For example, in some embodiments, including some embodiments of formula (V), the antisense oligomer conjugates of the present disclosure are according to formula (VA): [ka] wherein each Nu is a nucleobase that, together, forms a targeting sequence that is complementary to an exon 45 annealing site in the dystrophin pre-mRNA designated as H45A(-03+19).

[0290] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I). In various embodiments, each Nu from 1 to 30 and 5' to 3' is: [Table 7] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is [ka] It is.

[0291] In some embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (V) and formula (VA), the targeting sequence is 5'-CAATGCCATCCTGGAGTTCCTG-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO: 28). In various embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (V) and formula (VA), the targeting sequence is 5'-CAATGCCATCCTGGAGTTCCTG-3' (SEQ ID NO: 27).

[0292] In some embodiments, including for example, embodiments of the antisense oligomer conjugates of formula (I), the antisense oligomer conjugates of the present disclosure have the formula (VI): [ka] or a pharma- ceutically acceptable salt thereof.

[0293] In some embodiments, the antisense oligomer conjugate of formula (VI) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0294] In some embodiments, including for example, embodiments of the antisense oligomer conjugates of formula (VI), the antisense oligomer conjugates of the present disclosure are according to formula (VIA). [ka]

[0295] In some embodiments of the present disclosure, including some embodiments of the antisense oligomer conjugate of formula (I) and embodiments of the antisense oligomer conjugate of formula (VI), the antisense oligomer conjugate has the formula (VIB): [ka] or a pharma- ceutically acceptable salt thereof.

[0296] In some embodiments, the antisense oligomer conjugate of formula (VIB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt.

[0297] In some embodiments, including for example, the embodiment of the antisense oligomer conjugate of formula (VIB), the antisense oligomer conjugates of the present disclosure are according to formula (VIC). [ka]

[0298] For example, in some embodiments, including some embodiments of Formula (I), the antisense oligomer conjugate of the present disclosure has the formula (VII): [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each Nu is a nucleobase that together form a targeting sequence that is complementary to an exon 53 annealing site in the dystrophin pre-mRNA designated as H53A(+36+60).

[0299] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I). In various embodiments, each Nu from 1 to 25 and 5' to 3' is: [Table 8] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is independently [ka] In some embodiments, the antisense oligomer conjugate of formula (VII) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt. For example, in some embodiments, including some embodiments of formula (VII), the antisense oligomer conjugate of the present disclosure is according to formula (VIIA): [ka] wherein each Nu is a nucleobase that, taken together, forms a targeting sequence that is complementary to an exon 53 annealing site in the dystrophin pre-mRNA designated as H53A(+36+60).

[0300] In some embodiments, each Nu is independently selected from cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I). In various embodiments, each Nu from 1 to 25 and 5' to 3' is: [Table 9] Here, A is [ka] and C is [ka] and G is [ka] and X is [ka] In certain embodiments, each X is [ka] It is.

[0301] In some embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (VII) and formula (VIIA), the targeting sequence is 5'-GTTGCCTCCGGTTCTGAAGGTGTTC-3', where each thymine (T) is optionally a uracil (U) (SEQ ID NO: 30). In various embodiments, including, for example, embodiments of the antisense oligomer conjugates of formula (VII) and formula (VIIA), the targeting sequence is 5'-GTTGCCTCCGGTTCTGAAGGTGTTC-3' (SEQ ID NO: 29).

[0302] In some embodiments, including for example, embodiments of the antisense oligomer conjugates of formula (I), the antisense oligomer conjugates of the present disclosure have the formula (VIII): [ka] or a pharma- ceutically acceptable salt thereof.

[0303] In some embodiments, the antisense oligomer conjugate of formula (VIII) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt. In some embodiments, including, for example, embodiments of the antisense oligomer conjugate of formula (VIII), the antisense oligomer conjugate of the present disclosure is according to formula (VIIIA). [ka]

[0304] In some embodiments of the present disclosure, including some embodiments of the antisense oligomer conjugate of Formula (I) and embodiments of the antisense oligomer conjugate of Formula (VIII), the antisense oligomer conjugate has the formula (VIIIB): [ka] or a pharma- ceutically acceptable salt thereof.

[0305] In some embodiments, the antisense oligomer conjugate of formula (VIIIB) is its HCl (hydrochloric acid) salt. In certain embodiments, the HCl salt is a .6HCl salt. In some embodiments, including for example, embodiments of the antisense oligomer conjugate of formula (VIIIB), the antisense oligomer conjugate of the present disclosure is according to formula (VIIIC). [ka]

[0306] In one aspect, the disclosure provides an antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, capable of binding to a selected target and inducing exon skipping in a human dystrophin gene, the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, comprising a sequence of bases that is complementary to an exon 51 target region of a dystrophin pre-mRNA designated as an annealing site, the sequence of bases and the annealing site being selected from one of the following: [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] It is.

[0307] In one embodiment, the base sequence and annealing sites are selected from one of the following: [Table 11-1] [Table 11-2] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] and U is [ka] where Gm is a methylated guanine, Am is a methylated adenine, and mC is [ka] It is.

[0308] In another aspect, the present disclosure provides an antisense oligomer of formula (IX): [ka] or a pharma- ceutically acceptable salt thereof, wherein each Nu from 1 to n and from 5' to 3' corresponds to a nucleobase in the following sequence: [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] It is.

[0309] In one embodiment, the base sequence and annealing sites are selected from one of the following: [Table 13-1] [Table 13-2] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] and U is [ka] where Gm is a methylated guanine, Am is a methylated adenine, and mC is [ka] It is.

[0310] In another aspect, the present disclosure provides an antisense oligomer of formula (X): [ka] or a pharma- ceutically acceptable salt thereof, wherein each Nu from 1 to (n+1) and 5' to 3' corresponds to a nucleobase in the following sequence: [Table 14-1] [Table 14-2] [Table 14-3] [Table 14-4] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] It is.

[0311] In one embodiment, the base sequence and annealing sites are selected from one of the following: [Table 15-1] [Table 15-2] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] and U is [ka] where Gm is a methylated guanine, Am is a methylated adenine, and mC is [ka] It is.

[0312] In another aspect, the disclosure provides an antisense oligomer of formula (XA): [ka] In the formula, each Nu from 1 to (n+1) and 5' to 3' corresponds to a nucleic acid base in the following sequence: [Table 16-1] [Table 16-2] [Table 16-3] [Table 16-4] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] It is.

[0313] In one embodiment, the base sequence and annealing sites are selected from one of the following: [Table 17-1] [Table 17-2] Here, A is [ka] and C is [ka] and G is [ka] and T is [ka] and U is [ka] where Gm is a methylated guanine, Am is a methylated adenine, and mC is [ka] It is.

[0314] Nucleobase Modifications and Substitutions In certain embodiments, the antisense oligomer conjugates of the present disclosure are composed of RNA nucleobases and DNA nucleobases (often simply referred to in the art as "bases"). RNA bases are commonly known as adenine (A), uracil (U), cytosine (C), and guanine (G). DNA bases are commonly known as adenine (A), thymine (T), cytosine (C), and guanine (G). In various embodiments, the antisense oligomer conjugates of the present disclosure are composed of cytosine (C), guanine (G), thymine (T), adenine (A), 5-methylcytosine (5mC), uracil (U), and hypoxanthine (I).

[0315] In certain embodiments, one or more RNA or DNA bases in an oligomer may be modified or replaced with a base other than an RNA or DNA base. Oligomers containing modified or substituted bases include oligomers in which one or more of the purine or pyrimidine bases most commonly found in nucleic acids are replaced with a less common or unnatural base.

[0316] Purine bases contain a pyrimidine ring fused to an imidazole ring, as described by the general formula below: [ka]

[0317] Adenine and guanine are the two most commonly found purine nucleobases in nucleic acids. Other naturally occurring purines include N 6 -Methyladenine, N 2 Examples of methylguanine include, but are not limited to, -methylguanine, hypoxanthine, and 7-methylguanine.

[0318] The pyrimidine base contains a six-membered pyrimidine ring, as described by the general formula: [ka]

[0319] Cytosine, uracil, and thymine are the most commonly found pyrimidine bases in nucleic acids. Other naturally occurring pyrimidines include, but are not limited to, 5-methylcytosine, 5-hydroxymethylcytosine, pseudouracil, and 4-thiouracil. In one embodiment, the oligomer described herein contains a thymine base instead of uracil.

[0320] Other suitable bases include 2,6-diaminopurine, orotic acid, agmatidine, lysidine, 2-thiopyrimidines (e.g., 2-thiouracil, 2-thiothymine), G-clamps and derivatives thereof, 5-substituted pyrimidines (e.g., 5-halouracil, 5-propynyluracil, 5-propynylcytosine, 5-aminomethyluracil, 5-hydroxymethyluracil, 5-aminomethylcytosine, 5-hydroxymethylcytosine, Super T), 7-deazaguanine, 7-deazaadenine, 7-aza-2,6-diaminopurine, 8-aza-7-deazaguanine, 8-aza-7-deazaadenine, 8-aza-7-deaza-2,6-diaminopurine, Super G, Super A, and N4-ethylcytosine, or derivatives thereof; 2 -Cyclopentylguanine (cPent-G), N 2 -cyclopentyl-2-aminopurine (cPent-AP), and N 2-propyl-2-aminopurine (Pr-AP), pseudouracil, or derivatives thereof; and degenerate or universal bases such as 2,6-difluorotoluene, or non-existent bases such as base moieties (e.g., 1-deoxyribose, 1,2-dideoxyribose, 1-deoxy-2-O-methylribose; or pyrrolidine derivatives in which the ring oxygen is replaced with nitrogen (azaribose)). Examples of Super A, Super G, and Super T derivatives can be found in U.S. Pat. No. 6,683,173 (Epoch Biosciences), which is incorporated herein by reference in its entirety. cPent-G, cPent-AP, and Pr-AP have been shown to reduce immunostimulatory effects when incorporated into siRNA (Peacock H. et al. J. Am. Chem. Soc. 2011, 133, 9200). Pseudouracil is a naturally occurring isomerized version of uracil, with the C-glycoside rather than the normal N-glycoside in uridine. Pseudouridine-containing synthetic mRNAs may have an improved safety profile compared to uridine-containing mPvNAs (WO2009 / 127230, incorporated herein by reference in its entirety).

[0321] Certain nucleobases are particularly useful for increasing the binding affinity of the antisense oligomer conjugates of the present disclosure. These include 5-substituted pyrimidines, 6-azapyrimidines, and N-2, N-6, and O-6 substituted purines, including 2-aminopropyladenine, 5-propynyluracil, and 5-propynylcytosine. 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2°C, and are currently the more preferred base substitutions, particularly when combined with 2'-O-methoxyethyl sugar modifications. Additional exemplary modified nucleobases include those in which at least one hydrogen atom of the nucleobase is replaced with fluorine.

[0322] Pharmaceutically acceptable salts of the antisense oligomer conjugates Certain embodiments of the antisense oligomer conjugates described herein may contain basic functional groups, such as amino or alkylamino, and thus may form pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term "pharmaceutically acceptable salts" in this respect refers to relatively non-toxic inorganic and organic acid addition salts of the antisense oligomer conjugates of the present disclosure. These salts may be prepared in situ during the administration vehicle or dosage form manufacturing process, or may be prepared by separately reacting the purified antisense oligomer conjugates of the present disclosure in their free base form with a suitable organic or inorganic acid, and then isolating the salts thus formed during subsequent purification. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulfonate (see, e.g., Berge et al. (1977) "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0323] Pharmaceutically acceptable salts of the subject antisense oligomer conjugates include conventional non-toxic salts or quaternary ammonium salts of the antisense oligomer conjugates, for example, from non-toxic organic or inorganic acids. For example, such conventional non-toxic salts include those derived from inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, and the like; as well as salts prepared from organic acids, such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid (salicyclic acid), sulfanilic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, isothionic acid, and the like.

[0324] In certain embodiments, the antisense oligomer conjugates of the present disclosure may contain one or more acidic functional groups, and thus may form pharma-ceutically acceptable salts with pharma-ceutically acceptable bases.In these cases, the term "pharma-ceutically acceptable salts" refers to the relatively non-toxic inorganic and organic base addition salts of the antisense oligomer conjugates of the present disclosure.These salts may also be prepared in situ during the administration vehicle or dosage form manufacturing process, or may be prepared by separately reacting the purified antisense oligomer conjugate in its free acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharma-ceutically acceptable metal cation, with ammonia, or with a pharma-ceutically acceptable organic primary, secondary, or tertiary amine.Representative alkali or alkaline earth salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, and the like.Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like. (See, e.g., Berge et al., supra).

[0325] Formulations and Modes of Administration In certain embodiments, the present disclosure provides a formulation or pharmaceutical composition suitable for therapeutic delivery of the antisense oligomer conjugates described herein. Pharmaceutical formulations comprising antisense oligomers (e.g., PPMOs) conjugated to cell membrane-permeable peptides for DMD are described, for example, in U.S. Pat. No. 10,888,578, the disclosure of which is incorporated herein by reference. In certain embodiments, the present disclosure provides a pharma- ceutically acceptable composition comprising a therapeutically effective amount of one or more of the antisense oligomer conjugates described herein, formulated with one or more pharma- ceutically acceptable carriers (additives) and / or diluents. Although it is possible to administer the antisense oligomer conjugates of the present disclosure alone, it is preferred to administer the antisense oligomer conjugates as a pharmaceutical formulation (composition). In one embodiment, the antisense oligomer conjugate of the formulation is according to formula (I).

[0326] In another aspect, the present disclosure provides a pharmaceutical composition comprising an antisense oligomer of the present disclosure, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier, in some embodiments, the pharma- ceutically acceptable carrier is saline, including phosphate buffer.

[0327] The phrase "pharmacologically acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the subject being treated therewith.

[0328] As used herein, the phrase "pharmaceutically acceptable carrier" refers to non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material, or any type of formulation auxiliary.Some examples of materials that can serve as pharmaceutically acceptable carriers include, according to the formular's judgment, sugars such as lactose, glucose, sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, Olive oil, corn oil, and soybean oil; glycols, such as propylene glycol esters, for example, ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free distilled water; isotonic saline; Ringer's solution; ethyl alcohol; phosphate buffers; non-toxic compatible lubricants, such as sodium lauryl sulfate and magnesium stearate; colorants; release agents; coating agents; sweetening agents; flavoring agents; fragrances; preservatives; and antioxidants.

[0329] Methods for delivery of nucleic acid molecules that may be applicable to the antisense oligomer conjugates of the present disclosure are described, for example, in Akhtar et al., 1992, Trends Cell Bio., 2:139; Delivery Strategies for Antisense Oligonucleotide Therapeutics, ed. Akhtar, 1995, CRC Press, and Sullivan et al., PCT WO94 / 02595. These and other protocols can be utilized for the delivery of virtually any nucleic acid molecule, including the antisense oligomer conjugates of the present disclosure.

[0330] The pharmaceutical compositions of the present disclosure may be specially formulated for administration in solid or liquid form, including those adapted for: (1) oral administration, e.g., as an immersion (aqueous or non-aqueous solution or suspension), tablet (buccal, sublingual, or systemic absorption), bolus, powder, granule, paste for application to the tongue; (2) parenteral administration, e.g., by subcutaneous, intramuscular, intravenous, or epidural injection, e.g., as a sterile solution or suspension or sustained release formulation; (3) topical application, e.g., as a cream, ointment, or controlled release patch or spray for application to the skin; (4) vaginal or rectal administration, e.g., as a pessary, cream, or foam; (5) sublingual administration; (6) ocular administration; (7) transdermal administration; or (8) nasal administration.

[0331] Some examples of materials that can serve as pharma- ceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) glycols, such as protease, glycerol, sorbitol ... Examples of suitable non-toxic and non-toxic materials include, but are not limited to, propylene glycol, (11) polyols, such as glycerin, sorbitol, mannitol, and polyethylene glycol, (12) esters, such as ethyl oleate and ethyl laurate, (13) agar, (14) buffers, such as magnesium hydroxide and aluminum hydroxide, (15) alginic acid, (16) pyrogen-free distilled water, (17) isotonic saline, (18) Ringer's solution, (19) ethyl alcohol, (20) pH buffers, (21) polyesters, polycarbonates, and / or polyanhydrides, and (22) other non-toxic and compatible materials used in pharmaceutical formulations.

[0332] Additional non-limiting examples of drugs suitable for formulation with the antisense oligomer conjugates of the present disclosure include PEG-conjugated nucleic acids; phospholipid-conjugated nucleic acids; nucleic acids containing lipophilic moieties; phosphorothioates; P-glycoprotein inhibitors (such as Pluronic® P85) that can enhance drug entry into various tissues; biodegradable polymers, such as poly(D,L-lactide-coglycolide) microspheres for sustained delivery after implantation (Emerich, DF et al., 1999, Cell Transplant, 8, 47-58) Alkermes, Inc. Cambridge, Mass.; and loaded nanoparticles, such as those made of polybutyl cyanoacrylate, which can deliver drugs across the blood-brain barrier and alter neural uptake mechanisms (Prog Neuropsychopharmacol Biol Psychiatry, 23, 941-949, 1999).

[0333] The present disclosure also features the use of compositions comprising surface-modified liposomes containing poly(ethylene glycol) ("PEG") lipids (PEG-modified, branched, and unbranched, or combinations thereof, or long-circulating or stealth liposomes). The oligomeric conjugates of the present disclosure can also contain covalently attached PEG molecules of various molecular weights. These formulations provide a method for increasing drug accumulation in target tissues. This class of drug carriers resists opsonization and elimination by the mononuclear cell phagocytic system (MPS or RES), thereby allowing for extended blood circulation time and enhanced tissue exposure of encapsulated drugs (Lasic et al. Chem. Rev. 1995, 95, 2601-2627; Ishiwata et al., Chem. Pharm. Bull. 1995, 43, 1005-1011). Such liposomes have been shown to selectively accumulate in tumors, likely due to extravasation and entrapment in vascularized target tissues (Lasic et al., Science 1995, 267, 1275-1276; Oku et al., 1995, Biochim. Biophys. Acta, 1238, 86-90). Long-circulating liposomes enhance the pharmacokinetics and pharmacodynamics of DNA and RNA compared to conventional cationic liposomes, which are known to accumulate in tissues, particularly in MPS (Liu et al., J. Biol. Chem. 1995, 42, 24864-24870; Choi et al., International PCT Publication No. WO 96 / 10391; Ansell et al., International PCT Publication No. WO 96 / 10390; Holland et al., International PCT Publication No. WO 96 / 10392). Long-circulating liposomes also likely protect drugs from nuclease degradation to a greater extent than cationic liposomes, based on their ability to avoid accumulation in metabolically aggressive MPS tissues such as the liver and spleen.

[0334] In further embodiments, the present disclosure includes antisense oligomer conjugate compositions prepared for delivery as described in U.S. Patent Nos. 6,692,911, 7,163,695, and 7,070,807. In this regard, in one embodiment, the present disclosure provides an antisense oligomer conjugate of the present disclosure in a composition comprising a copolymer of lysine and histidine (HK) (described in U.S. Patent Nos. 7,163,695, 7,070,807, and 6,692,911), either alone or in combination with PEG (e.g., branched or unbranched PEG, or a mixture of both), in combination with PEG and a targeting moiety, or in combination with a crosslinker. In certain embodiments, the present disclosure provides an antisense oligomer conjugate in a pharmaceutical composition comprising gluconate-modified polyhistidine or gluconyl-polyhistidine / transferrin-polylysine. One of skill in the art will also recognize that amino acids with properties similar to His and Lys can be substituted within the compositions.

[0335] Wetting agents, emulsifying agents, and lubricants (such as sodium lauryl sulfate and magnesium stearate), coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservatives, and antioxidants can also be present in the composition.

[0336] Examples of pharma- ceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine ​​hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite, and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediaminetetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0337] Methods of preparing these formulations or pharmaceutical compositions include the step of bringing into association an antisense oligomer conjugate of the present disclosure with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association an antisense oligomer conjugate of the present disclosure with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

[0338] Formulations of the disclosure suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (with a flavored base, usually sucrose and acacia or tragacanth), powder, granules, or as a solution or suspension in an aqueous or non-aqueous solution, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as a pastille (with an inert base, e.g., gelatin and glycerin, or sucrose and acacia) and / or as a mouthwash, each containing a predetermined amount of an antisense oligomer conjugate of the disclosure as an active ingredient. The antisense oligomer conjugates of the disclosure may also be administered as a bolus, electuary, or paste.

[0339] Pharmaceutical compositions suitable for parenteral administration may contain one or more oligomer conjugates of the present disclosure in combination with one or more pharma- ceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders that can be reconstituted immediately before use into sterile injectable solutions or dispersions, which may contain sugars, alcohols, antioxidants, buffers, bacteriostatic agents, solutes that render the formulation isotonic with the blood of the intended recipient, or suspending or thickening agents. Examples of suitable aqueous and non-aqueous carriers that may be used in the pharmaceutical compositions of the present disclosure include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, etc.), and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Proper fluidity may be maintained, for example, by the use of coating materials such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants. In one embodiment, the antisense oligomer conjugate of the pharmaceutical composition is according to formula (I).

[0340] These pharmaceutical compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents, and dispersing agents. Prevention of the action of microorganisms on the subject oligomeric conjugates may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like, in the compositions. Furthermore, prolonged absorption of the injectable dosage form may be brought about by the inclusion of agents which delay absorption, such as aluminum monostearate and gelatin.

[0341] In some cases, it is desirable to delay the absorption of drugs from subcutaneous or intramuscular injections in order to prolong the effect of drugs.This can be achieved by using liquid suspensions of crystalline or amorphous materials with poor water solubility, among other methods known in the art.The rate of absorption of the drug then depends on its dissolution rate, which in turn can depend on crystal size and crystalline form.Alternatively, delayed absorption of parenterally administered drug forms can be achieved by dissolving or suspending the drug in an oil vehicle.

[0342] Injectable depot forms can be made by forming microencapsule matrices of the subject oligomer conjugates in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of oligomer to polymer and the nature of the particular polymer used, the rate of oligomer release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissues.

[0343] When the antisense oligomer conjugates of the present disclosure are administered as pharmaceuticals to humans and animals, they may be administered per se or as a pharmaceutical composition containing, for example, 0.1-99% (more preferably 10-30%) of the antisense oligomer conjugate in combination with a pharma- ceutically acceptable carrier.

[0344] In certain embodiments, the antisense oligomer conjugate is administered in a liquid pharmaceutical formulation, and the concentration of the conjugate is about 50 mg / ml.

[0345] Regardless of the route of administration selected, the antisense oligomer conjugates of the present disclosure, which may be used in a suitable hydrated form, and / or the pharmaceutical compositions of the present disclosure, may be formulated into pharma- ceutically acceptable dosage forms by conventional methods known to those skilled in the art. Actual dosage levels of the active ingredients in the pharmaceutical compositions of the present disclosure may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being unacceptably toxic to the patient.

[0346] How to use Treatment with antisense oligomer CPP conjugates can be administered to patients in need thereof using the dosing regimens described in this disclosure.

[0347] In one aspect, the disclosure provides a method for treating DMD in a subject in need thereof, the subject having a mutation in the dystrophin gene that is amenable to exon skipping, the method comprising administering to the subject an antisense oligomeric CPP conjugate according to a dosing regimen described herein. In some aspects, the exon is exon 44, exon 45, exon 50, exon 51, exon 52, or exon 53 of the human dystrophin gene.

[0348] In another aspect, the disclosure provides a method of restoring the mRNA reading frame to induce dystrophin production in a subject having a mutation in the dystrophin gene amenable to exon skipping (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53 skipping), the method comprising administering to the subject an antisense oligomeric CPP conjugate according to a dosing regimen described herein.

[0349] In another aspect, the present disclosure provides a method of excluding exons (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53) from a dystrophin pre-mRNA during mRNA processing in a subject with a mutation in the dystrophin gene that is amenable to exon skipping, the method comprising administering to the subject an antisense oligomeric CPP conjugate according to a dosing regimen described herein. In another aspect, the present disclosure provides a method of combining exons (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53) of a dystrophin pre-mRNA in a subject with a mutation in the dystrophin gene that is amenable to exon skipping (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53 skipping), the method comprising administering to the subject an antisense oligomeric CPP conjugate according to a dosing regimen described herein.

[0350] In another aspect, the disclosure provides a method of targeting exposure levels of an antisense oligomeric CPP conjugate in a DMD patient having a mutation amenable to exon skipping (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53 skipping), the method comprising administering to a subject an antisense oligomeric CPP conjugate according to a dosing regimen described herein.

[0351] In another aspect, the disclosure provides a method of reducing the severity and / or frequency of hypomagnesemia in a patient being treated with an antisense oligomeric CPP conjugate, the method comprising administering to the subject an antisense oligomeric CPP conjugate and a magnesium supplement according to the dosing regimen described herein.

[0352] In another aspect, the disclosure provides a method for alleviating one or more symptoms of DMD in a human patient, the method comprising administering to the patient an antisense oligomeric CPP conjugate according to the dosing regimen described herein, the patient undergoing steroid therapy. In some aspects, the conjugate is administered to induce exon skipping (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53 of the human dystrophin gene). In some aspects, administration of the conjugate according to the dosing regimen described herein and steroid therapy are initiated simultaneously. In other aspects, the patient undergoes steroid therapy prior to administration of the antisense oligomeric CPP conjugate according to the dosing regimen described herein. In this aspect, the steroid is preferably administered at least one day, more preferably at least one week, more preferably at least two weeks, more preferably at least three weeks prior to administration of the antisense oligomeric CPP conjugate according to the dosing regimen described herein. In one particular embodiment, the prior steroid treatment is for a period of at least three weeks prior to administration of the antisense oligomeric CPP conjugate according to the dosing regimen described herein.

[0353] In some embodiments, the steroid is a glucocorticosteroid. The glucocorticosteroid may be selected from, for example, prednisone, dexamethasone, prednisolone, or deflazacort. The dose range of the steroid (e.g., glucocorticosteroid) used in the therapeutic applications described herein is designed based on dose escalation studies in clinical trials with strict protocol requirements. The usual dose is about 0.5-1.0 mg / kg / day, preferably about 0.75 mg / kg / day for prednisone and prednisolone, and about 0.4-1.4 mg / kg / day, preferably about 0.9 mg / kg / day for deflazacort.

[0354] The term "restoration" with respect to dystrophin synthesis or production generally refers to the production of dystrophin protein, including truncated forms of dystrophin, in a muscular dystrophy patient following treatment with an antisense oligomer conjugate described herein. In some embodiments, the treatment results in an increase of 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% (including all integers therebetween) of new dystrophin production in the patient. In some embodiments, the treatment increases the number of dystrophin-positive fibers in the subject to at least about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, or about 95% to 100% of normal. In other embodiments, the treatment increases the number of dystrophin-positive fibers in the subject to about 20% to about 60% or about 30% to about 50% of normal. The percentage of dystrophin-positive fibers in a patient after treatment can be determined by muscle biopsy using known techniques. For example, a muscle biopsy can be taken from a suitable muscle, such as the patient's biceps.

[0355] Analysis of the percentage of positive dystrophin fibers can be performed before and / or after treatment, or at time points throughout the course of treatment. In some embodiments, the post-treatment biopsy is taken from the muscle contralateral to the pre-treatment biopsy. Pre-treatment and post-treatment dystrophin expression analysis can be performed using any suitable assay for dystrophin. In some embodiments, immunohistochemical detection is performed on tissue sections from muscle biopsies using an antibody that is a marker for dystrophin, such as a monoclonal or polyclonal antibody. For example, the MANDYS106 antibody, which is a sensitive marker for dystrophin, can be used. Any suitable secondary antibody can be used.

[0356] In some embodiments, the percentage of dystrophin-positive fibers is calculated by dividing the number of positive fibers by the total number of fibers counted. A normal muscle sample has 100% dystrophin-positive fibers. Thus, the percentage of dystrophin-positive fibers can be expressed as a percentage of normal. When counting dystrophin-positive fibers in post-treatment muscle, a baseline can be set using a pre-treatment muscle section from the patient to control for the presence of trace levels of dystrophin in pre-treatment muscle as well as revertant fibers. This can be used as a threshold for counting dystrophin-positive fibers in post-treatment muscle sections of that patient. In other embodiments, antibody-stained tissue sections can also be used for dystrophin quantification using Bioquant image analysis software (Bioquant Image Analysis Corporation, Nashville, TN). Total dystrophin fluorescent signal intensity can be reported as a percentage of normal. Additionally, Western blot analysis using monoclonal or polyclonal anti-dystrophin antibodies can be used to determine the percentage of dystrophin-positive fibers. For example, the anti-dystrophin antibody NCL-Dys1 from Leica Biosystems can be used. The percentage of dystrophin-positive fibers can also be analyzed by determining the expression of components of the sarcoglycan complex (β, γ) and / or neuronal NOS.

[0357] In some embodiments, treatment with the antisense oligomer conjugates of the present disclosure slows or reduces the progressive respiratory muscle dysfunction and / or failure in DMD patients that would be expected without treatment. In some embodiments, treatment with the antisense oligomer conjugates of the present disclosure can reduce or eliminate the need for ventilatory support that would be expected without treatment. In some embodiments, measurements of respiratory function to track disease progression and evaluate potential therapeutic interventions include maximum inspiratory pressure (MIP), maximum expiratory pressure (MEP), and forced vital capacity (FVC). MIP and MEP measure the pressure levels a person can generate during inhalation and exhalation, respectively, and are sensitive measures of respiratory muscle strength. MIP is a measure of diaphragm muscle weakness.

[0358] In some embodiments, MEP may decline before changes in other pulmonary function tests, including MIP and FVC. In certain embodiments, MEP may be an early indicator of respiratory dysfunction. In certain embodiments, FVC may be used to measure the total volume of air expelled during forced expiration after maximal inspiration. In DMD patients, FVC increases in parallel with physical growth until the early teens. However, as growth slows or becomes poorer with disease progression and muscle weakness progresses, vital capacity enters a decline phase, declining at an average rate of about 8-8.5 percent per year after age 10-12. In certain embodiments, percent predicted MIP (MIP adjusted for weight), percent predicted MEP (MEP adjusted for age), and percent predicted FVC (FVC adjusted for age and height) are supportive analyses.

[0359] As used herein, the terms "subject" and "patient" include any animal that exhibits or is at risk of exhibiting a condition that can be treated with the antisense oligomer conjugates of the present disclosure, such as a subject (or patient) that has or is at risk of having DMD or BMD, or any of the symptoms associated with these conditions (e.g., muscle fiber loss). Suitable subjects (or patients) include laboratory animals (such as mice, rats, rabbits, or guinea pigs), farm animals, and household animals or pets (such as cats or dogs). Non-human primates, and preferably human patients (or subjects) are included. Also included are methods of producing dystrophin in subjects (or patients) that have a mutation in the dystrophin gene that is amenable to exon skipping (e.g., exon 44, exon 45, exon 50, exon 51, exon 52, exon 53 skipping).

[0360] As used herein, the phrases "systemic administration," "administered systemically," "peripheral administration," and "administered peripherally" refer to administration of a compound, drug, or other material other than directly into the central nervous system, thereby entering the patient's system and thus being subject to metabolic and other similar processes, such as, for example, subcutaneous administration.

[0361] The phrase "targeting sequence" refers to a sequence of nucleobases of an oligomer that is complementary to a sequence of nucleotides in a target pre-mRNA. In some embodiments of the present disclosure, the sequence of nucleotides in the target pre-mRNA is an exon 51 annealing site in a dystrophin pre-mRNA designated as H51A (+66 + 95). In some embodiments of the present disclosure, the sequence of nucleotides in the target pre-mRNA is an exon 45 annealing site in a dystrophin pre-mRNA designated as H45A (-03 + 19). In some embodiments, the sequence of nucleotides in the target pre-mRNA is an exon 53 annealing site in a dystrophin pre-mRNA designated as H53A (+36 + 60).

[0362] "Treatment" of a subject (e.g., a mammal, such as a human) is any type of intervention used in an attempt to change the natural course of the subject. Treatment includes, but is not limited to, administration of an antisense oligomer conjugate or a pharmaceutical composition thereof, and can be performed either prophylactically or after the onset of a pathological event or contact with a pathogen. Treatment includes any desired effect on the symptoms or pathology of a disease or condition associated with a dystrophin protein, such as in certain forms of muscular dystrophy, and may include, for example, a minimal change or improvement in one or more measurable markers of the disease or condition being treated. Also included are "prophylactic" treatments, which may be aimed at slowing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset. "Treatment" or "prevention" does not necessarily indicate a complete eradication, cure, or prevention of the disease or condition or its associated symptoms.

[0363] In some embodiments, treatment with an antisense oligomeric CPP conjugate of the present disclosure increases new dystrophin production, slows disease progression, slows or reduces lameness, reduces muscle inflammation, reduces muscle damage, improves muscle function, reduces loss of lung function, and / or enhances muscle regeneration that would be expected without treatment. In some embodiments, treatment maintains, slows, or slows disease progression. In some embodiments, treatment maintains ambulation or reduces lameness. In some embodiments, treatment maintains lung function or reduces loss of lung function. In some embodiments, treatment maintains or increases the patient's stable walking distance, for example, as measured by the 6-minute walk test (6MWT). In some embodiments, treatment maintains or reduces the time to walk / run 10 meters (i.e., 10-meter walk / run test). In some embodiments, treatment maintains or reduces the time to stand up from a supine position (i.e., time to stand test). In some embodiments, the treatment maintains or reduces the time to climb four standard stairs (i.e., the four-step climb test). In some embodiments, the treatment maintains or reduces inflammation in the patient's muscles, for example, as measured by MRI (e.g., MRI of leg muscles). In some embodiments, the MRI measures T2 and / or fat fraction to identify muscle degeneration. MRI can identify changes in muscle structure and composition caused by inflammation, edema, muscle damage, and fat infiltration.

[0364] In some embodiments, treatment with the antisense oligomer conjugates of the present disclosure increases new dystrophin production and delays or reduces the loss of ambulation that would be expected without treatment. For example, treatment may stabilize, maintain, improve, or increase the walking ability of a subject (e.g., stabilization of ambulation). In some embodiments, treatment maintains or increases the patient's stable walking distance, as measured, for example, by the 6-minute walk test (6MWT) described by McDonald, et al. (Muscle Nerve, 2010; 42: 966-74, incorporated herein by reference). The change in 6-minute walk distance (6MWD) may be expressed as an absolute value, a percentage change, or a change in % predicted value. In some embodiments, treatment maintains or improves stable walking distance in the 6MWT from a 20% deficit in the subject compared to healthy counterparts. The performance of a DMD patient in the 6MWT compared to the typical performance of a healthy counterpart may be determined by calculating the % predicted value. For example, the % predicted 6MWD for men may be calculated using the following equation: 196.72+(39.81×age)−(1.36×age) 2 ) + (132.28 × height in meters). For women, the % predicted 6MWD is calculated using the following equation: 188.61 + (51.50 × age) - (1.86 × age 2 ) + (86.10 × height in meters) (Henricson et al. PLoS Curr., 2012, version 2, incorporated herein by reference). In some embodiments, treatment with an antisense oligomer increases stable walking distance in a patient from baseline to greater than 3, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, or 50 meters (including all integers therebetween).

[0365] Loss of muscle function in DMD patients may occur against the background of normal childhood growth and development. Indeed, children with DMD may show an increase in walking distance during the 6MWT over the course of about a year, despite progressive muscle impairment. In some embodiments, 6MWD from DMD patients is compared to existing normative data from typically developing control subjects, as well as age- and sex-matched subjects. In some embodiments, normal growth and development may be accounted for using an equation based on age and height fitted to normative data. Using such an equation, 6MWD in subjects with DMD may be converted to percent predicted (% predicted) values. In certain embodiments, analysis of % predicted 6MWD data represents a way to account for normal growth and development, and may show that increases in function at younger ages (e.g., 7 years or younger) represent a stabilization of performance rather than an improvement in performance in DMD patients (Henricson et al. PLoS Curr., 2012, version 2, incorporated herein by reference).

[0366] In order to distinguish between different antisense molecules, a nomenclature system for antisense molecules has been proposed and published (see Mann et al., (2002) J Gen Med 4, 644-654). This nomenclature system has become particularly suitable when testing several slightly different antisense molecules all directed against the same target region, as shown below. H#A / D(x:y)

[0367] The first letter indicates the species (e.g., H: human, M: mouse, C: dog). "#" indicates the target dystrophin exon number. "A / D" indicates the acceptor or donor splice site at the beginning and end of the exon, respectively. (xy) represents the annealing coordinates, and "-" or "+" indicates the intron or exon sequence, respectively. For example, A(-6+18) indicates the last 6 bases of the intron preceding the target exon and the first 18 bases of the target exon. The closest splice site will be the acceptor, so these coordinates are preceded by "A". The description of the annealing coordinates at the donor splice site can be D(+2-18), where the last 2 exon bases and the first 18 intron bases correspond to the annealing site of the antisense molecule. The entire exon annealing coordinate is represented by A(+65+85), ie, the site from 65th to 85th nucleotides from the start of the exon.

[0368] Restoring the dystrophin reading frame using exon skipping Potential therapeutic approach for the treatment of DMD caused by out-of-frame mutation of dystrophin gene is suggested by milder form of dystrophinopathy known as BMD caused by in-frame mutation.The ability to convert out-of-frame mutation into in-frame mutation hypothetically maintains mRNA reading frame and produces internally truncated but functional dystrophin protein.The antisense oligomer conjugate of the present disclosure is designed to achieve this.

[0369] Clinical outcomes to analyze the effect of antisense oligomer conjugates complementary to a target region of human dystrophin pre-mRNA and inducing exon skipping include percentage of dystrophin positive fibers (PDPF), 6 minute walk test (6MWT), loss of ambulation (LOA), North Star Ambulation Assessment (NSAA), pulmonary function tests (PFTs), ability to stand up (from supine position) without external support, de novo dystrophin production, and other functional measures.

[0370] In some embodiments, the present disclosure provides a method for producing dystrophin in a subject having a mutation in the dystrophin gene that is amenable to exon skipping (e.g., exons 44, 45, 50, 51, 52, 53), the method comprising administering to the subject an antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof, as described herein. In certain embodiments, the present disclosure provides a method for restoring the mRNA reading frame to induce dystrophin protein production in a subject having Duchenne muscular dystrophy (DMD) having a mutation in the dystrophin gene that is amenable to exon skipping (e.g., exons 44, 45, 50, 51, 52, 53). Protein production can be measured by reverse transcription polymerase chain reaction (RT-PCR), Western blot analysis, or immunohistochemistry (IHC).

[0371] In some embodiments, the disclosure provides a method for treating DMD in a subject in need thereof, the subject having a mutation in the dystrophin gene that is amenable to exon skipping (e.g., exons 44, 45, 50, 51, 52, 53), the method comprising administering to the subject an antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof, as described herein. In various embodiments, treatment of the subject is measured by delaying disease progression. In some embodiments, treatment of the subject is measured by maintenance of ambulation in the subject, or reduction of ambulation in the subject. In some embodiments, ambulation is measured using the 6 Minute Walk Test (6MWT). In certain embodiments, ambulation is measured using the North Start Ambulation Assessment (NSAA).

[0372] In various embodiments, the present disclosure provides a method for maintaining lung function or reducing loss of lung function in a subject with DMD, the subject having a mutation in the DMD gene that is amenable to exon skipping (e.g., exons 44, 45, 50, 51, 52, 53), the method comprising administering to the subject an antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof as described herein. In some embodiments, the lung function is measured as maximum expiratory pressure (MEP). In certain embodiments, the lung function is measured as maximum inspiratory pressure (MIP). In some embodiments, the lung function is measured as forced vital capacity (FVC).

[0373] In certain aspects, the methods of the disclosure comprise administering to a subject having DMD a pharmaceutical formulation comprising an antisense oligomer conjugate described herein, or a pharma- ceutically acceptable salt thereof, wherein the concentration of the conjugate in the formulation is about 50 mg / ml.

[0374] In certain embodiments, the antisense oligomer conjugate described herein is described for use in therapy.In certain embodiments, the antisense oligomer conjugate described herein is described for use in the treatment of Duchenne muscular dystrophy.In certain embodiments, the antisense oligomer conjugate described herein is described for use in the manufacture of a medicament for use in therapy.In certain embodiments, the antisense oligomer conjugate described herein is described for use in the manufacture of a medicament for the treatment of Duchenne muscular dystrophy. EXAMPLES

[0375] Although the above disclosure has been described in some detail by way of illustration and examples for purposes of clarity of understanding, it will be readily apparent to one skilled in the art, in light of the teachings of the present disclosure, that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims. The following examples are provided by way of illustration only, and not by way of limitation. Those skilled in the art will readily recognize a variety of non-critical parameters that can be changed or modified to produce substantially similar results.

[0376] Example 1. Toxicity and Toxicokinetic Studies of PPMO-1 in Non-Human Primates Study 1. Toxicity and toxicokinetics study of 12 weeks of intravenous infusion (once every 4 weeks) in cynomolgus monkeys with a 4 week recovery period. Non-human primates (NHPs) received three 1-h IV infusions of vehicle control or PPMO-1 at dose levels of 30 or 60 mg / kg on days 1, 29, and 57. PPMO-1 is an antisense oligo CPP conjugate having the following structure: [ka]

[0377] Blood samples were collected pre-dose and 1, 2, 4, 8, 12, 16, and 24 hours post-infusion for plasma PK analysis after the first and third doses. Mean body weight was 5.99 ± 0.84 Kg (n = 7) on day 1, range 5.1 to 7.5 Kg.

[0378] Study 2. Non-GLP (Laboratory Laboratory Practice) pharmacokinetic (PK), pharmacodynamic, and renal safety evaluation of PPMO-1 following four repeated intravenous doses once every four weeks (Q4W) in cynomolgus monkeys. NHPs received four 1-h IV infusions of vehicle control or PPMO-1 at dose levels of 30 or 60 mg / kg on days 1, 29, 57, and 85. Blood samples were collected pre-dose and at 1, 2, 4, 8, 12, 16, and 24 hours post-infusion for plasma PK analysis after the first and fourth doses. Mean body weight was 2.25 ± 0.1 Kg (n = 8) on day 1, range 2.1-2.4 Kg.

[0379] As shown in FIG. 1, animals in Study 1 had higher initial body weights than animals in Study 2.

[0380] As shown in FIG. 2, animals in Study 1 had a higher plasma AUC (AUC last (h*μg / mL) and C max (μg / kg) exposure. First and last dose PK were used for analysis.

[0381] There is a relationship of dose-normalized PK to body weight as shown in Figure 3. Higher body weight is associated with higher PK, both AUC and Cmax.

[0382] Example 2. Development of a new dosing paradigm to achieve target drug exposure in DMD patients A correlation between body weight and plasma exposure to PPMO-1 was observed in ongoing clinical trials. As shown in Figure 4, the C of PPMO-1 at 20 mg / kg in Study 5051-102 max was 2.6-fold higher than the 20 mg / kg dose in Study 5051-201. max The difference was related to the significant difference in body weight, 55.9±17.1 kg vs. 33.0±16.5 kg (mean±SD) in 5051-102. Indeed, at 20 mg / kg and 30 mg / kg, C maxA good correlation was observed between dose and body weight (Figure 5). This suggests that a weight-based fixed mg / kg approach for a broad DMD patient population with a body weight range of about 10 to about 100 kg may result in apparently variable drug exposure and heterogeneous outcomes of treatment effects on efficacy and safety. For example, for patients with lower body weight, drug exposure may be below the optimal target exposure range, resulting in suboptimal efficacy or even no efficacy, while for patients with higher body weight, drug exposure may be above the optimal target exposure range, resulting in potential safety issues.

[0383] Therefore, a weight range approach has been developed to place DMD patients in a certain weight range, and different doses are selected within each weight range to achieve targeted drug exposure. Target exposures are defined based on clinical data on exposure-response relationships for efficacy and safety. A population pharmacokinetic model was developed based on existing clinical data for SRP-5051, and simulations were performed to inform weight range and dose selection to achieve targeted exposure across the DMD population. A similar method is described in Model-Based Approach for Optimization of Atazanir Dose Recommendations for HIV-Infected Pediatric Patients (Hong, Y. et al, Antimicrobial Agents and Chemotherapy, Dec. 2011, p. 5746-5752).

[0384] Two target exposures of area under the plasma concentration versus time curve (AUC) were selected: a lower target exposure with a median of 150 μg / mL*h and a range of 100-200 μg / mL*h, and a higher target exposure with a median of 300 μg / mL*h and a range of 200-500 μg / mL*h. The body weight across the DMD population can be divided into different ranges (1, 2, 3, or more) and a uniform fixed mg dose can be selected within each body weight range. For the higher target exposure, as shown in Table 1B, If the number of weight ranges is 1, then a flat dose of 1200mg is expected to achieve the target exposure across body weights of 15-100kg. If the number of weight ranges is 2, then 15-40 kg is selected as the first range with a flat dose of 1100 mg, and 40-100 kg is selected as the second range with a flat dose of 1300 mg. If the number of weight ranges is 3, then 15-25 kg is selected as the first range with a flat dose of 1100 mg, 25-50 kg is selected as the second range with a flat dose of 1200 mg, and 15-25 kg is selected as the third range with a flat dose of 1400 mg. The more weight ranges selected, the lower the variability in drug exposure is expected. For patients weighing less than 15 kg, a different dose may be selected based on the pharmacokinetic properties. Overall, this demonstrates that a weight range approach (one or more range numbers, and a flat dose within each range, e.g., 900-1800 mg) can adequately achieve higher target exposures.

[0385] Similarly, Table 2A shows the weight range approach for the lower target exposures.

[0386] In summary, this demonstrates that the weight range approach (one or more range numbers and flat doses within each range) can adequately achieve target lower and higher exposures. [Table 2-1] [Table 2-2]

[0387] Example 3. Clinical Administration Rationale Clinical and non-clinical findings suggest that higher drug exposure is associated with higher body weight, which was consistently observed among boys with DMD, between heavier adult healthy volunteers and boys with DMD, and in non-clinical studies, raising challenges in achieving consistent and less variable drug exposure in a diverse and broad population of children with DMD.

[0388] To avoid potential overdosing for heavier weight patients and underdosing for lighter weight patients, a population PK model-informed weight-stratified dosing regimen has been implemented, with the targeted DMD population in the study placed into two weight strata (18-50 kg, and >50 kg) with a flat dose within each stratum to achieve the target exposure.

[0389] The two target exposures for the study are selected by fixing the drug plasma exposures in patients who had previously undergone muscle biopsy at 20 and 30 mg / kg, respectively, as follows: Lower target exposure with AUC of 1.180 μg h / mL (range: 120-240 μg h / mL) to bracket the exposure at 20 mg / kg for biopsied patients (geometric mean AUC 128 μg h / mL, N=2) and the exposure at 30 mg / kg for biopsied patients. Based on the relationship between dystrophin expression and AUC, an AUC of 180 μg h / mL appears to be associated with an approximately 5% increase in dystrophin protein levels from baseline. 2. Higher target exposure at AUC of 300 μg h / mL (range: 200-500 μg h / mL), which mimics the exposure at 30 mg / kg for biopsied patients (AUC geometric mean 310 μg h / mL, range 184-504 μg h / mL, N=4). Based on the relationship between dystrophin expression and AUC, an AUC of 300 μg h / mL appears to be associated with a 5% or greater increase in dystrophin protein levels from baseline.

[0390] Thus, these two target exposures differ by approximately 1.7-fold with slight overlap in AUC (overlapping AUCs for 200–240 μg h / mL), which is a reasonable difference in exposure separation to assess the optimal balance between benefit and risk. The proposed target exposures are supported by existing nonclinical safety data. The higher target AUC (300 μg h / mL) is approximately 1.6-, 2.7-, and 1.3-fold lower than the NOAEL of 60 mg / kg in the SR-19-051 (NHP non-GLP), SR-20-027 (NHP GLP), and SR-20-028 (juvenile rat GLP) studies, respectively. For the lower target AUC, a higher safety margin exists, approximately 2.7-, 4.6-, and 2.2-fold lower, respectively.

[0391] As shown in Table 3, to achieve a high target exposure of 300 μg·h / mL, lower doses (total mg) are proposed for lower weight strata: 1250 mg for ≥18 kg to <50 kg, and 1350 mg for ≥50 kg. The expected 95% prediction interval for AUC is well within the target range (200–500 μg·h / mL), specifically, the upper boundary is up to 420 μg·h / mL (well below 500 μg·h / mL), which may provide additional protection for safety while still providing adequate dystrophin expression. A similar scenario has been proposed to target a lower target exposure.

[0392] In summary, a weight-stratified dosing regimen targeting the two exposures maximizes the opportunity to achieve an optimal benefit-risk ratio. [Table 3]

[0393] In addition to the embodiments described above, the following embodiments are also contemplated by the present disclosure.

[0394] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (IV), as described above, wherein the conjugate is administered at a dose effective to provide a mean area under the curve (AUC) of the conjugate or a pharmacologic acceptable salt thereof that is about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0395] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (IV), as described above, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 300 mg to about 1500 mg of the 6HCl salt of the conjugate to achieve a mean AUC of about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0396] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL. In another embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 120 to about 240 ug·h / mL. In another embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 200 to about 500 ug·h / mL.

[0397] In certain embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate. In certain other embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate.

[0398] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 300 mg to about 700 mg of the 6HCl salt of the conjugate.

[0399] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg of the 6HCl salt of the conjugate.

[0400] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 900 mg to about 1200 mg of the 6HCl salt of the conjugate.

[0401] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 1100 mg of the 6HCl salt of the conjugate.

[0402] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 750 mg to about 1250 mg of the 6HCl salt of the conjugate.

[0403] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 600 mg to about 800 mg of the 6HCl salt of the conjugate.

[0404] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 650 mg of the 6HCl salt of the conjugate.

[0405] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1000 mg to about 1300 mg of the 6HCl salt of the conjugate.

[0406] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1200 mg of the 6HCl salt of the conjugate.

[0407] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg in a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0408] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 1250 mg of the 6HCl salt of the conjugate.

[0409] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 700 mg to about 900 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0410] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0411] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1200 mg to about 1500 mg of the 6HCl salt of the conjugate.

[0412] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1400 mg of the 6HCl salt of the conjugate.

[0413] In one embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 850 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0414] In another embodiment, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1350 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0415] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (VI), as described above, wherein the conjugate is administered at a dose effective to provide a mean area under the curve (AUC) of the conjugate or a pharmaceutical acceptable salt thereof that is about 100 to about 200 ug·h / mL, 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0416] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (VI), as described above, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 300 mg to about 1500 mg of the 6HCl salt of the conjugate to achieve a mean AUC of about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0417] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL. In another embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 120 to about 240 ug·h / mL. In another embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 200 to about 500 ug·h / mL.

[0418] In certain embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate. In certain other embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate.

[0419] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 300 mg to about 700 mg of the 6HCl salt of the conjugate.

[0420] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg of the 6HCl salt of the conjugate.

[0421] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 900 mg to about 1200 mg of the 6HCl salt of the conjugate.

[0422] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 1100 mg of the 6HCl salt of the conjugate.

[0423] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0424] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 1250 mg of the 6HCl salt of the conjugate.

[0425] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 25 kg to about 50 kg at a dose equivalent to about 600 mg to about 800 mg of the 6HCl salt of the conjugate.

[0426] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 650 mg of the 6HCl salt of the conjugate.

[0427] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1000 mg to about 1300 mg of the 6HCl salt of the conjugate.

[0428] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1200 mg of the 6HCl salt of the conjugate.

[0429] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 700 mg to about 900 mg of the 6HCl salt of the conjugate.

[0430] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg or greater at a dose equivalent to about 750 mg to about 950 mg of the 6HCl salt of the conjugate.

[0431] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1250 mg to about 1500 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0432] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 50 kg to about 100 kg at a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0433] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1200 mg to about 1500 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

[0434] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1400 mg of the 6HCl salt of the conjugate.

[0435] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 850 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0436] In one embodiment, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 1350 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg or greater.

[0437] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (VIII), as described above, wherein the conjugate is administered at a dose effective to provide a mean area under the curve (AUC) of the conjugate or a pharmaceutical acceptable salt thereof that is about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0438] In one embodiment, the disclosure relates to a method of treating a Duchenne muscular dystrophy (DMD) patient having a mutation amenable to exon skipping, the method comprising administering to the patient an antisense oligomer conjugate of Formula (VIII), as described above, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 300 mg to about 1500 mg of the 6HCl salt of the conjugate to achieve a mean AUC of about 100 to about 200 ug·h / mL, about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0439] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL. In another embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 120 to about 240 ug·h / mL. In another embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 200 to about 500 ug·h / mL.

[0440] In certain embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate. In certain other embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate.

[0441] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 300 mg to about 700 mg of the 6HCl salt of the conjugate.

[0442] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg of the 6HCl salt of the conjugate.

[0443] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 900 mg to about 1200 mg of the 6HCl salt of the conjugate.

[0444] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 1100 mg of the 6HCl salt of the conjugate.

[0445] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0446] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 18 kg to about 50 kg at a dose equivalent to about 1250 mg of the 6HCl salt of the conjugate.

[0447] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 25 kg to about 50 kg at a dose equivalent to about 600 mg to about 800 mg of the 6HCl salt of the conjugate.

[0448] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 650 mg of the 6HCl salt of the conjugate.

[0449] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1000 mg to about 1300 mg of the 6HCl salt of the conjugate.

[0450] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg at a dose equivalent to about 1200 mg of the 6HCl salt of the conjugate.

[0451] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 700 mg to about 900 mg of the 6HCl salt of the conjugate.

[0452] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 50 kg to about 100 kg at a dose equivalent to about 750 mg of the 6HCl salt of the conjugate.

[0453] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1200 mg to about 1500 mg of the 6HCl salt of the conjugate.

[0454] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing about 50 kg to about 100 kg at a dose equivalent to about 1400 mg of the 6HCl salt of the conjugate.

[0455] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing more than about 50 kg at a dose equivalent to about 850 mg of the 6HCl salt of the conjugate.

[0456] In one embodiment, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing greater than about 50 kg at a dose equivalent to about 1350 mg of the 6HCl salt of the conjugate.

[0457] One embodiment of the present disclosure relates to a method of targeting exposure levels of an antisense oligomer conjugate of Formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, in a patient having Duchenne muscular dystrophy (DMD) having a mutation amenable to exon 51 skipping, the method comprising administering to the patient the conjugate or a pharma- ceutically acceptable salt thereof, at a dose equivalent to about 300 mg to about 900 mg of the .6HCl salt of the conjugate, once every four weeks, to achieve an exposure level corresponding to an AUC of about 100 to about 200 ug·h / mL.

[0458] One embodiment of the present disclosure relates to a method of targeting exposure levels of an antisense oligomer conjugate of Formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, in a patient having Duchenne muscular dystrophy (DMD) having a mutation amenable to exon 51 skipping, the method comprising administering to the patient the conjugate or a pharma- ceutically acceptable salt thereof, at a dose equivalent to about 700 mg to about 900 mg of the .6HCl salt of the conjugate, once every four weeks, to achieve an exposure level corresponding to an AUC of about 120 to about 240 ug·h / mL.

[0459] One embodiment of the present disclosure relates to a method of targeting exposure levels of an antisense oligomer conjugate of Formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, in a patient having Duchenne muscular dystrophy (DMD) having a mutation amenable to exon 51 skipping, the method comprising administering to the patient the conjugate or a pharma- ceutically acceptable salt thereof, at a dose equivalent to about 800 mg to about 1400 mg of the .6HCl salt of the conjugate, once every four weeks, to achieve an exposure level corresponding to an AUC of about 200 to about 500 ug·h / mL.

[0460] One embodiment of the present disclosure relates to a method of targeting exposure levels of an antisense oligomer conjugate of Formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, in a patient having Duchenne muscular dystrophy (DMD) having a mutation amenable to exon 51 skipping, the method comprising administering to the patient the conjugate or a pharma- ceutically acceptable salt thereof, at a dose equivalent to about 950 mg to about 1500 mg of the .6HCl salt of the conjugate, once every four weeks, to achieve an exposure level corresponding to an AUC of about 200 to about 500 ug·h / mL.

[0461] One embodiment of the present disclosure relates to a method of targeting exposure levels of an antisense oligomeric conjugate of formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, in a patient having Duchenne muscular dystrophy (DMD) with a mutation amenable to exon 51 skipping, the method comprising administering to the patient a dose of 100 mg / mL of an antisense oligomeric conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof according to the following schedule: i) about 300 mg to about 700 mg or about 900 mg to about 1,200 mg once every 4 weeks for a patient weighing about 10 kg to about 25 kg; ii) about 600 mg to about 800 mg or about 1000 mg to about 1300 mg once every 4 weeks for a patient weighing about 25 kg to about 50 kg, or iii) about 700 mg to about 900 mg or about 1200 mg to about 1500 mg once every 4 weeks for a patient weighing about 50 kg to about 100 kg; to achieve an exposure level corresponding to an AUC that is about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0462] One embodiment of the present disclosure relates to a method of treating DMD in a patient, the method comprising administering to the patient an antisense oligomer conjugate of formula (IV) as described above, or a pharma- ceutically acceptable salt thereof, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered according to the following schedule: i) about 500 mg to about 700 mg or about 1000 mg to about 1200 mg once every 4 weeks for a patient weighing about 15 kg to about 40 kg, or ii) about 600 mg to about 800 mg or about 1100 mg to about 1400 mg once every 4 weeks for a patient weighing about 40 kg to about 100 kg; to achieve a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0463] One embodiment of the present disclosure relates to a method of treating DMD in a patient, the method comprising administering to the patient an antisense oligomer conjugate of formula (VI) as described above, or a pharma- ceutically acceptable salt thereof, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered according to the following schedule: i) about 500 mg to about 700 mg or about 1000 mg to about 1200 mg once every 4 weeks for a patient weighing about 15 kg to about 40 kg, or ii) about 600 mg to about 800 mg or about 1100 mg to about 1400 mg once every 4 weeks for a patient weighing about 40 kg to about 100 kg; to achieve a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0464] One embodiment of the present disclosure relates to a method of treating DMD in a patient, the method comprising administering to the patient an antisense oligomer conjugate of formula (VIII) as described above, or a pharma- ceutically acceptable salt thereof, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered according to the following schedule: i) about 500 mg to about 700 mg or about 1000 mg to about 1200 mg once every 4 weeks for a patient weighing about 15 kg to about 40 kg, or ii) about 600 mg to about 800 mg or about 1100 mg to about 1400 mg once every 4 weeks for a patient weighing about 40 kg to about 100 kg; to achieve a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0465] One embodiment of the present disclosure relates to a method of treating DMD in a patient, the method comprising administering to the patient an antisense oligomer conjugate of formula (VIII) as described above, or a pharma- ceutically acceptable salt thereof, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered according to the following schedule: iii) about 700 mg to about 800 mg or about 1200 mg to about 1250 mg once every 4 weeks for a patient weighing about 18 kg to about 50 kg, or iv) about 800 mg to about 900 mg or about 1300 mg to about 1400 mg once every 4 weeks for a patient weighing about 40 kg to about 100 kg; to achieve a mean AUC of about 120 to about 240 ug·h / mL, or about 200 to about 500 ug·h / mL.

[0466] One embodiment of the disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, the method comprising administering to the patient: i) an antisense oligomeric conjugate of formula (IV); ii) a magnesium supplement; and The conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 20 mg / kg, about 30 mg / kg, or about 40 mg / kg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium levels.

[0467] One embodiment of the disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, the method comprising administering to the patient: iii) an antisense oligomeric conjugate of formula (IV), iv) magnesium supplements,

[0468] In some embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered in an amount of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 1300 mg, about 500 mg to about 1000 mg, The magnesium supplement is administered at a dose equivalent to about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0469] In some embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0470] In some embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0471] In some embodiments, the conjugate of formula (IV) or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 20 mg / kg, about 30 mg / kg, or about 40 mg / kg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0472] One embodiment of the present disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, the method comprising administering to the patient: v) an antisense oligomeric conjugate of formula (VI); vi) magnesium supplements,

[0473] In some embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered in an amount of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 1300 mg, about 500 mg to about 1000 mg, The magnesium supplement is administered at a dose equivalent to about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0474] In some embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0475] In some embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0476] In some embodiments, the conjugate of formula (VI) or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 20 mg / kg, about 30 mg / kg, or about 40 mg / kg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0477] One embodiment of the disclosure relates to a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, the method comprising administering to the patient: vii) an antisense oligomeric conjugate of formula (VIII): viii) magnesium supplements,

[0478] In some embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 1300 mg, or about 500 mg to about 1000 mg. , about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0479] In some embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0480] In some embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered at a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize the patient's serum magnesium level. In one embodiment, the conjugate is administered once every four weeks. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

[0481] In some embodiments, the conjugate of formula (VIII) or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 20 mg / kg, about 30 mg / kg, or about 40 mg / kg of the 6HCl salt of the conjugate, and the magnesium supplement is administered at a dose sufficient to normalize serum magnesium levels in the patient. In one embodiment, the magnesium supplement is administered at a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

Claims

1. 1. A composition for treating a patient having Duchenne muscular dystrophy (DMD) with a mutation amenable to exon skipping in a patient in need thereof, comprising an antisense oligomeric conjugate of formula (I): 【Chemistry 140】 or a pharma- ceutically acceptable salt thereof, During the ceremony, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 141】 is a moiety selected from R １００ is a cell membrane penetrating peptide, R ２００ is hydrogen, R １ is C １ -C ６ is alkyl, 20. A composition comprising: a) a first concentration of said conjugate, said second concentration of said conjugate being administered at a dose effective to provide a mean AUC of said conjugate or said pharmacologic salt thereof, which is about 100 to about 200 ug.h / mL, or is about 200 to about 500 ug.h / mL.

2. 1. A composition for treating a patient having Duchenne muscular dystrophy (DMD) with a mutation suitable for exon 51 skipping in a patient in need thereof, comprising an antisense oligomeric conjugate of formula (I): 【Chemistry 142】 or a pharma- ceutically acceptable salt thereof, During the ceremony, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 143】 is a moiety selected from R １００ is a cell membrane penetrating peptide, R ２００ is hydrogen, R １ is C １ -C ６ is alkyl, the conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks at a dose equivalent to about 300 mg to about 1500 mg of the 6HCl salt of the conjugate; A composition characterized by achieving a mean AUC of about 100 to about 200 ug·h / mL, or about 200 to about 500 ug·h / mL.

3. The conjugate or a pharma- ceutically acceptable salt thereof is administered in an amount of about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 1300 mg, about 500 mg to about 1000 mg 3. The composition of claim 1 or 2, wherein the 600 mg to about 900 mg of the 6HCl salt of the conjugate is administered in a dose equivalent to about 1000 mg, about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the 6HCl salt of the conjugate.

4. The composition of any one of claims 1 to 3, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose effective to provide a mean AUC of about 100 to about 200 ug·h / mL.

5. The composition of any one of claims 1 to 3, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered at a dose effective to provide a mean AUC of about 200 to about 500 ug·h / mL.

6. 5. The composition of claim 1, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate.

7. 6. The composition of any one of claims 1 to 3 or 5, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate.

8. The composition of any one of claims 1 to 7, wherein the cell membrane permeable peptide is selected from RXRRXRRXRRXR (SEQ ID NO: 18), RFFRFFRFFR (SEQ ID NO: 19), RXRRXRRXRRXRXB (SEQ ID NO: 20), RFFRFFRFFRG (SEQ ID NO: 21), RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), R is arginine, X is 6-aminohexanoic acid, B is β-alanine, F is phenylalanine, and G is glycine.

9. The composition according to any one of claims 1 to 8, wherein the cell membrane penetrating peptide is selected from RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), wherein R is arginine and G is glycine.

10. The composition according to any one of claims 1 to 9, wherein the cell membrane penetrating peptide is RRRRRRG (SEQ ID NO: 22), in which R is arginine and G is glycine.

11. The composition of any one of claims 1 to 10, wherein the antisense oligomer conjugate induces exon skipping in the human dystrophin gene.

12. The composition of any one of claims 1 to 11, wherein the exon is selected from exons 44, 45, 50, 51, 52, or 53.

13. The composition of any one of claims 1 to 12, wherein the exon is selected from exon 45, 51, or 53.

14. The antisense oligomer conjugate has formula (III): 【Chemistry 144】 or a pharma- ceutically acceptable salt thereof; 14. The composition of any one of claims 1 to 13, wherein each Nu is a nucleobase that together form a targeting sequence that is complementary to an exon annealing site in a dystrophin pre-mRNA.

15. The antisense oligomer conjugate has formula (IV): 【Chemistry 145】 or a pharma- ceutically acceptable salt thereof.

16. The composition of any one of claims 1 to 15, wherein the antisense oligomer conjugate has the formula (IVA). 【Chemistry 146】

17. The antisense oligomer conjugate has the formula (VI): 【Chemistry 147】 or a pharma- ceutically acceptable salt thereof.

18. The composition of any one of claims 1 to 11 or 17, wherein the antisense oligomer conjugate has the formula (VIA). 【Chemistry 148】

19. The antisense oligomer conjugate has the formula (VIII): 【Chemistry 149】 or a pharma- ceutically acceptable salt thereof.

20. The composition of any one of claims 1 to 14 or 19, wherein the antisense oligomer conjugate has the formula (VIIIA): 【Chemistry 150】

21. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg to about 700 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 10 kg to about 25 kg.

22. 22. The composition of any one of claims 1 to 21, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 600 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 10 kg to about 25 kg.

23. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 900 mg to about 1200 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 10 kg to about 25 kg.

24. 24. The composition of any one of claims 1 to 20 or 23, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1100 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 10 kg to about 25 kg.

25. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 600 mg to about 800 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 25 kg to about 50 kg.

26. 26. The composition of any one of claims 1 to 20 or 25, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 650 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 25 kg to about 50 kg.

27. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1000 mg to about 1300 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 25 kg to about 50 kg.

28. 28. The composition of any one of claims 1 to 20 or 27, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1200 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 25 kg to about 50 kg.

29. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 700 mg to about 900 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing about 50 kg to about 100 kg.

30. 30. The composition of any one of claims 1 to 20 or 29, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 750 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 50 kg to about 100 kg.

31. 21. The composition of any one of claims 1 to 20, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1200 mg to about 1500 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 50 kg to about 100 kg.

32. 32. The composition of any one of claims 1 to 20 or 31, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 1400 mg of the 6HCl salt of the conjugate once every four weeks to a patient weighing from about 50 kg to about 100 kg.

33. 1. A composition for treating a patient having Duchenne muscular dystrophy (DMD) with a mutation suitable for exon 51 skipping in a patient in need thereof, comprising an antisense oligomeric conjugate of formula (I): 【Chemistry 151】 or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 152】 is a moiety selected from R １００ is a cell membrane penetrating peptide, R ２００ is hydrogen, R １ is C １ -C ６ alkyl), and the antisense oligomer conjugate, or a pharma- ceutically acceptable salt thereof, comprises Schedule below: i) about 300 mg to about 700 mg or about 900 mg to about 1200 mg once every 4 weeks for a patient weighing about 10 kg to about 25 kg; ii) about 600 mg to about 800 mg or about 1000 mg to about 1300 mg once every 4 weeks for a patient weighing about 25 kg to about 50 kg; or iii) about 700 mg to about 900 mg or about 1200 mg to about 1500 mg once every 4 weeks for a patient weighing about 50 kg to about 100 kg; when administered to said patient at a dose equivalent to the dose of the 6HCl salt of said conjugate according to the method of claim 1, to achieve a mean AUC that is about 100 to about 200 ug h / mL, or about 200 to about 500 ug h / mL.

34. 34. The composition of claim 33, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 10 kg to about 25 kg at a dose equivalent to about 600 mg or about 1100 mg of the 6HCl salt of the conjugate.

35. 34. The composition of claim 33, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 25 kg to about 50 kg in a dose equivalent to about 650 mg or about 1200 mg of the 6HCl salt of the conjugate.

36. 34. The composition of claim 33, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered once every four weeks to a patient weighing from about 50 kg to about 100 kg in a dose equivalent to about 750 mg or about 1400 mg of the 6HCl salt of the conjugate.

37. The composition according to any one of claims 1 to 36, characterized in that the composition is administered to the patient in combination with a magnesium supplement.

38. 1. A combination for use in a method of treating a patient with Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate comprising: iii) An antisense oligomeric conjugate of formula (I): 【Chemistry 153】 or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 154】 is a moiety selected from R １００ is a cell membrane penetrating peptide, R ２００ is hydrogen, R １ is C １ -C ６ alkyl), and iv) a magnesium supplement, wherein said method comprises administering said combination to said patient.

39. The conjugate or a pharma- ceutically acceptable salt thereof is about 300 mg to about 1200 mg, about 300 mg to about 1000 mg, about 300 mg to about 750 mg, about 300 mg to about 500 mg, about 400 mg to about 1400 mg, about 400 mg to about 1100 mg, about 400 mg to about 900 mg, about 400 mg to about 600 mg, about 500 mg to about 1500 mg, about 500 mg to about 1300 mg, about 500 mg to about 1000 mg 39. The combination of claim 38, wherein the 6HCI salt of the conjugate is administered in a dose equivalent to about 500 mg to about 950 mg, about 500 mg to about 850 mg, about 500 mg to about 750 mg, about 500 mg to about 650 mg, about 600 mg to about 1400 mg, about 600 mg to about 1200 mg, about 600 mg to about 1200 mg, about 600 mg to about 1000, or about 600 mg to about 900 mg of the 6HCI salt of the conjugate.

40. 40. The combination of claim 38 or 39, wherein the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 300 mg, about 350 mg, about 400 mg, about 450 mg, about 500 mg, about 550 mg, about 600 mg, about 650 mg, about 700 mg, about 750 mg, about 800 mg, about 850 mg, or about 900 mg of the 6HCl salt of the conjugate.

41. 41. The combination according to any one of claims 38 to 40, characterized in that the conjugate or a pharma- ceutically acceptable salt thereof is administered in a dose equivalent to about 950 mg, about 1000 mg, about 1050 mg, about 1100 mg, about 1150 mg, about 1200 mg, about 1250 mg, about 1300 mg, about 1350 mg, about 1400 mg, about 1450 mg, or about 1500 mg of the 6HCl salt of the conjugate.

42. The combination according to any one of claims 38 to 41, wherein the cell membrane permeable peptide is selected from RXRRXRRXRRXR (SEQ ID NO: 18), RFFRFFRFFR (SEQ ID NO: 19), RXRRXRRXRRXRXB (SEQ ID NO: 20), RFFRFFRFFRG (SEQ ID NO: 21), RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), R is arginine, X is 6-aminohexanoic acid, B is β-alanine, F is phenylalanine, and G is glycine.

43. The combination according to any one of claims 38 to 42, wherein the cell membrane penetrating peptide is selected from RRRRRRG (SEQ ID NO: 22), RRRRRR (SEQ ID NO: 23), RRRRRG (SEQ ID NO: 24), or RRRRR (SEQ ID NO: 25), wherein R is arginine and G is glycine.

44. The combination according to any one of claims 38 to 43, wherein the cell membrane penetrating peptide is RRRRRRG (SEQ ID NO: 22), in which R is arginine and G is glycine.

45. The combination according to any one of claims 38 to 44, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof causes exon skipping in the human dystrophin gene.

46. The combination according to any one of claims 38 to 45, wherein the exon is selected from exons 44, 45, 50, 51, 52, or 53.

47. The combination according to any one of claims 38 to 46, wherein the exon is selected from exons 45, 51, or 53.

48. A combination according to any one of claims 38 to 47, characterized in that the magnesium supplement is administered in an amount effective to provide normal serum magnesium levels.

49. A combination according to any one of claims 38 to 48, characterized in that the magnesium supplement is administered in a dose equivalent to about 100 to about 2400 mg of magnesium oxide per day.

50. 50. The combination according to any one of claims 38 to 49, characterized in that the magnesium supplement is administered in a dose equivalent to about 400 to about 2400 mg of magnesium oxide per day.

51. A combination described in any one of claims 38 to 50, wherein the method further comprises measuring the patient's serum magnesium level at a time point of at least two weeks after the administration.

52. A combination described in any one of claims 38 to 51, wherein the method further comprises administering a second dose of a magnesium supplement, with the dose being based on the measured serum magnesium level.

53. 53. The combination of any one of claims 38 to 52, wherein the magnesium supplement is selected from magnesium oxide, magnesium citrate, magnesium carbonate, magnesium hydrogen phosphate, magnesium glycerophosphate, magnesium trisilicate, magnesium hydroxide, magnesium carbonate hydroxide, magnesium acetate, magnesium citrate, magnesium lactate, magnesium gluconate, magnesium chloride, magnesium aspartate, magnesium caprylate, magnesium ascorbate, magnesium taurate, magnesium malate, and magnesium diglycinate, magnesium pidulate, or magnesium sulfate.

54. A combination according to any one of claims 38 to 53, wherein the magnesium supplement is derived from magnesium oxide.

55. A combination for treating a patient with an antisense oligomer conjugate, comprising an antisense oligomer conjugate and a magnesium supplement, said combination being administered to a patient in need of said treatment.

56. 56. The combination of any one of claims 38 to 55, wherein the antisense oligomer conjugate or a pharma- ceutically acceptable salt thereof is administered in a pharmaceutical formulation, and the concentration of the conjugate in the formulation is about 50 mg / ml.

57. A composition for use in a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, comprising: Antisense oligomer conjugates of formula (I): 【Chemistry 153】 or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 154】 is a moiety selected from R 100 is a cell membrane penetrating peptide, R 200 is hydrogen, and R 1 is C 1 -C 6 alkyl; The method comprises administering to the patient the composition in combination with a magnesium supplement.

58. A composition for use in a method of treating a patient having Duchenne muscular dystrophy (DMD) with an antisense oligomer conjugate, comprising: and a magnesium supplement, the method comprising administering to the patient an antisense oligomer conjugate of formula (I): 【Chemistry 153】 or a pharma- ceutically acceptable salt thereof (In the formula, n is 1 to 40; each Nu is a nucleobase that together form a targeting sequence complementary to an exon annealing site in the dystrophin pre-mRNA, T' is 【Chemistry 154】 is a moiety selected from R 100 is a cell membrane permeable peptide, R 200 is hydrogen, and R 1 is C 1 -C 6 alkyl), comprising administering to said patient a compound comprising the steps of:

59. A composition for treating a patient with an antisense oligomer conjugate, comprising an antisense oligomer conjugate, the composition being administered to a patient in need of the treatment in combination with a magnesium supplement.

60. A composition for treating a patient with an antisense oligomer conjugate, comprising a magnesium supplement, the composition being administered in combination with the antisense oligomer conjugate to a patient in need of the treatment.

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