Cardiac-specific targeting-peptide (ctp), compositions, and uses thereof

Abstract

Disclosed is a synthetic, non-naturally occurring 12-amino acid peptide, Cardiac Targeting Peptide, belonging to the larger class of cell penetrating peptides, for delivery of both diagnostic and potentially therapeutic agents to the heart. Also disclosed are subsequent generations of this Cardiac Targeting Peptide with improved, higher efficiency of cardiac uptake for CTP-mediated delivery of antiarrhythmics subjects for treating Atrial fibrillation or Ventricular fibrillation, CTP-mediated delivery of neuregulin-1β for treating systolic heart failure (SHF), and Szeto-Schiller peptides for treatment of diastolic congestive heart failure, among other applications.

What is claimed is: 
     
         1 . A peptide comprising Cardiac-specific Targeting-Peptide (CTP) of the sequence of SEQ ID NO: 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 28, 29, or 34 linked to a moiety having antiarrhythmic properties. 
     
     
         2 . The peptide of  claim 1 , wherein the moiety is a class III antiarrhythmic compound or a class IV antiarrhythmic compound. 
     
     
         3 . The peptide of  claim 1 , wherein the moiety is amiodarone. 
     
     
         4 . The peptide of  claim 1 , wherein the CTP has sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 5. 
     
     
         5 . The peptide according to any one of  claims 1 - 4 , wherein the moiety having antiarrhythmic properties is linked to the CTP using Schiff base chemistry. 
     
     
         6 . The peptide of any one of  claims 1 - 5 , wherein the moiety having antiarrhythmic properties is linked upstream of the N-terminus of the CTP peptide. 
     
     
         7 . The peptide of any one of  claims 1 - 6 , wherein the peptide is optionally further connected to a label at the N- and/or the C-termini. 
     
     
         8 . The peptide of any one of  claims 1 - 6 , wherein the peptide is optionally further connected to a label at the C-termini. 
     
     
         9 . The peptide of  claim 7  or  8 , wherein the label is a green fluorescent moiety or a red fluorescent moiety. 
     
     
         10 . The peptide of  claim 9 , wherein the green florescent moiety is 6-carboxyfluorescein. 
     
     
         11 . The peptide of  claim 9 , wherein the red fluorescent moiety is Cy5.5. 
     
     
         12 . A formulation comprising a peptide according to any one of  claims 1 - 6  and a pharmaceutically acceptable carrier. 
     
     
         13 . The formulation of  claim 12  is a sustained-delivery formulation. 
     
     
         14 . The formulation of  claim 12  or  13 , wherein the formulation uses a controlled release system. 
     
     
         15 . The formulation of  claim 12  or  13 , wherein the formulation uses a slow release system. 
     
     
         16 . The formulation of  claim 12  or  13  delivers the peptide according to any one of  claims 1 - 6  over a period of at least 6 hours. 
     
     
         17 . The formulation of any one of  claims 12 - 16 , wherein the delivery is over a period of at least 12 hours. 
     
     
         18 . The formulation of any one of  claims 12 - 16 , wherein the delivery is over a period of at least 24 hours. 
     
     
         19 . The formulation of any one of  claims 12 - 16 , wherein the formulation is administered at least once daily. 
     
     
         20 . The formulation of any one of  claims 12 - 16 , wherein the formulation is administered at least once a week, once every two weeks, once a month, once every 30 days, once every 45 days, or once every 60 days. 
     
     
         21 . A method of introducing a moiety having antiarrhythmic properties into a cardiac muscle cell comprising administering to the cardiac muscle cell the peptide of any one of  claims 1 - 6  or a formulation of any one of  claims 12 - 20  in an amount effective to introduce the moiety having antiarrhythmic properties into the muscle cell. 
     
     
         22 . The method of  claim 21 , wherein the peptide further comprises a microsphere or nanoparticle. 
     
     
         23 . A method of treating a human subject suffering from atrial and/or ventricular arrhythmias, comprising administering to the human subject the peptide of any one of  claims 1 - 6  or a formulation comprising a peptide according to any one of  claims 1 - 5  according to any one of  claims 12 - 20 . 
     
     
         24 . The method of  claim 23 , wherein the ventricular arrhythmias is selected from premature ventricular contractions, ventricular tachycardia, and ventricular fibrillation. 
     
     
         25 . The method of  claim 23 , wherein the atrial arrhythmias is atrial fibrillation. 
     
     
         26 . A six amino acid Cardiac-specific Targeting-Peptide (CTP 6aa ) comprising the sequence of Xaa 1  Xaa 2  Y Xaa 3  Xaa 4  T (SEQ ID NO: 4). 
     
     
         27 . The CTP 6aa  of  claim 26  comprising the sequence of S Q Xaa 1  S R Xaa 2  (SEQ ID NO: 6). 
     
     
         28 . The CTP 6aa  of  claim 26 , wherein Xaa 1  is serine (S). 
     
     
         29 . The CTP 6aa  of  claim 26 , wherein Xaa 2  is glutamine (Q). 
     
     
         30 . The CTP 6aa  of  claim 26 , wherein Xaa 3  is serine (S). 
     
     
         31 . The CTP 6aa  of  claim 26 , wherein Xaa 4  is arginine (R). 
     
     
         32 . The CTP 6aa  of  claim 26 , wherein Xaa 1  and Xaa 2  are serine (S) and glutamine (Q), respectively. 
     
     
         33 . The CTP 6aa  of  claim 26 , wherein Xaa 1  and Xaa 3  are both serine (S). 
     
     
         34 . The CTP 6aa  of  claim 26 , wherein Xaa 1  and Xaa 4  are serine (S) and arginine (R), respectively. 
     
     
         35 . The CTP 6aa  of  claim 26 , wherein Xaa 2  and Xaa 3  are glutamine (Q) and serine (S), respectively. 
     
     
         36 . The CTP 6aa  of  claim 26 , wherein the CTP 6aa  comprises the sequence SQYSRT (SEQ ID NO: 5). 
     
     
         37 . The CTP 6aa  of  claim 27 , wherein Xaa 1  is alanine (A) and the CTP 6aa  comprises the sequence of SQASRXaa 2  (SEQ ID NO: 7), or optionally, Xaa 1  is tryptophan (W) and the CTP 6aa  comprises the sequence of SQWSRXaa 2  (SEQ ID NO: 8), or Xaa 1  is tyrosine (Y) and the CTP 6aa  comprises the sequence of SQYSRXaa 2  (SEQ ID NO: 9). 
     
     
         38 . The CTP 6aa  of  claim 27 , wherein Xaa 2  is threonine (T), and Xaa 1  is alanine (A), tryptophan (W), or tyrosine (Y) comprising the sequence of SQASRT (SEQ ID NO: 10), SQWSRT (SEQ ID NO: 11), or SQYSRT (SEQ ID NO: 5), respectively. 
     
     
         39 . The CTP 6aa  of  claim 27 , wherein Xaa 2  is alanine (A). 
     
     
         40 . The CTP 6aa  of  claim 27 , wherein Xaa 1  is tyrosine (Y) and Xaa 2  is alanine (A). 
     
     
         41 . The CTP 6aa  of  claim 27 , wherein the CTP 6aa  comprises the sequence SQYSRT (SEQ ID NO: 5). 
     
     
         42 . The CTP 6aa  of any one of  claims 26 - 41 , wherein the CTP 6aa  is conjugated or fused to a nanoparticle or a moiety having antiarrhythmic properties. 
     
     
         43 . A method of treating cardiac tissue or a cardiac condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising the conjugated or fused CTP 6aa  of  claim 42 . 
     
     
         44 . The CTP 6aa  of any one of  claims 26 - 41 , wherein the CTP 6aa  is conjugated or fused to a tafazzin peptide. 
     
     
         45 . The CTP 6aa  of  claim 44 , wherein the CTP 6aa  is conjugated or fused to the tafazzin peptide through a polypeptide linker. 
     
     
         46 . A method of treating a subject having a disorder associated with a tafazzin deficiency or a remodeled cardiolipin deficiency, comprising administering to the subject a therapeutically effective amount of a composition comprising the tafazzin-conjugated CTP 6aa  of  claim 44  or  45 . 
     
     
         47 . A method of treating a subject having, or at risk of developing Barth syndrome, comprising administering to the subject a therapeutically effective amount of a composition comprising the tafazzin-conjugated CTP 6aa  of  claim 44  or  45 . 
     
     
         48 . The CTP 6aa  of any one of  claims 26 - 41 , wherein the CTP 6aa  is conjugated to a detectable agent. 
     
     
         49 . The CTP 6aa  of  claim 48 , wherein the detectable agent is a radionuclide or radioactive moiety, biotin, luciferase, an enzyme, rhodamine, a fluorophore, nanoparticle, microbubbles, liposomes or a luminescent moiety. 
     
     
         50 . The CTP 6aa  of any one of  claims 26 - 41  formulated as a delivery vehicle/agent. 
     
     
         51 . The CTP 6aa  of  claim 50 , wherein the CTP 6aa  is conjugated to a drug or therapeutic, a nanoparticle, a peptide, a protein, or a detectable agent. 
     
     
         52 . The CTP 6aa  of  claim 51 , wherein the CTP 6aa  is conjugated to the drug or therapeutic, the nanoparticle, the peptide, the protein, or the detectable agent via an ester linkage, disulfide or protease sensitive linkers. 
     
     
         53 . The CTP 6aa  of  claim 51  or  52 , wherein the nanoparticle comprises a drug or therapeutic. 
     
     
         54 . The CTP 6aa  of any one of  claims 51 - 53 , wherein the drug or therapeutic is selected from an antibody or fragment thereof, SB239063, superoxide dismutase, HGF, FGF-1, FGF-2, an NF-κB inhibitor, NSD peptide, heme oxygenase, an antioxidant, iNOS, S100A1, catalase, glutathione peroxidase, a TGFβ inhibitor, VEGF, sonic hedgehog protein, HGF, an IAP, prostaglandin, Pyrvinium Pamoate, Diprotin A, Szeto-Schiller peptide, cyclosporine, and amiodarone. 
     
     
         55 . A method of treating cardiac tissue or a cardiac condition in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising the CTP 6aa  of  claim 54 . 
     
     
         56 . A method of introducing a cargo into a cardiac muscle cell comprising administering, to the cardiac muscle cell, an amount of a complex comprising the CTP 6aa  of any one of  claims 26 - 41  linked to a cargo effective to introduce the cargo into the muscle cell. 
     
     
         57 . The method of  claim 56 , wherein the cargo comprises a radioisotope, fluorescent marker, gadolinium marker, luciferase marker, microsphere or nanoparticle. 
     
     
         58 . A method of treating a human subject suffering from a myocardial infarction, comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the human subject, a therapeutically effective amount of a complex comprising the CTP 6aa  of any one of  claims 26 - 41  linked to a cargo, where the cargo inhibits cell death, inhibits arrhythmias, improves contractility, lengthens subject survival, or a combination thereof. 
     
     
         59 . The method of  claim 58 , wherein the cargo is selected from an NF-κB inhibitor, NSD peptide, heme oxygenase, an antioxidant, iNOS, S100A1, superoxide dismutase, catalase, glutathione peroxidase, a TGFβ inhibitor, VEGF, FGF-1, FGF-2, sonic hedgehog protein, HGF, a class III antiarrhythmic compound, a class IV antiarrhythmic compound, and an IAP. 
     
     
         60 . A method of treating a human subject suffering from a myocardial infarction, comprising introducing a cargo into a cardiac tissue of the human subject comprising administering, to the human subject, a therapeutically effective amount of a complex comprising the CTP 6aa  of any one of  claims 26 - 41  linked to a cargo, wherein the cargo inhibits cell death, inhibits arrhythmias, improves contractility, lengthens subject survival, or a combination thereof. 
     
     
         61 . The method of  claim 60 , wherein the cargo is selected from an NF-κB inhibitor, NBD peptide, heme oxygenase, an antioxidant, iNOS, S100A1, superoxide dismutase, catalase, glutathione peroxidase, a TGFβ inhibitor, VEGF, FGF-1, FGF-2, sonic hedgehog protein, HGF, a class III antiarrhythmic compound, a class IV antiarrhythmic compound, and an IAP. 
     
     
         62 . A method of treating a subject suffering from a metabolic defect that damages the heart comprising introducing a cargo into a cardiac muscle cell of the human subject comprising administering, to the subject, a therapeutically effective amount of a complex comprising the CTP 6aa  of any one of  claims 26 - 41  linked to a cargo, wherein the cargo corrects the metabolic defect. 
     
     
         63 . The method of  claim 62 , wherein the metabolic defect is Gaucher's disease and the cargo is glucocerebrosidase. 
     
     
         64 . A method of treating a subject suffering from a metabolic defect that damages the heart comprising introducing a cargo into a cardiac tissue of the human subject comprising administering, to the subject, a therapeutically effective amount of a complex comprising the CTP 6aa  of any one of  claims 26 - 41  linked to a cargo, where the cargo corrects the metabolic defect. 
     
     
         65 . The method of  claim 64 , where the metabolic defect is Gaucher's disease and the cargo is glucocerebrosidase. 
     
     
         66 . A peptide comprising Cardiac-specific Targeting-Peptide (CTP) of the sequence of SEQ ID NO: 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 28, 29, or 34 linked to an SS peptide. 
     
     
         67 . The peptide according to  claim 66 , wherein the SS peptide is linked upstream of the N-terminus of the CTP peptide. 
     
     
         68 . The peptide according to  claim 67 , comprising an ester linkage between the CTP and the SS peptide. 
     
     
         69 . The peptide according to  claim 68 , wherein the ester linkage is an ester linkage cleavable by an intracellular esterase. 
     
     
         70 . A peptide comprising Cardiac-specific Targeting-Peptide (CTP) of the sequence of SEQ ID NO: 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 28, 29, or 34 linked to heme-oxygenase 1 protein. 
     
     
         71 . The peptide of  claim 70 , wherein the heme-oxygenase 1 protein is linked upstream of the N-terminus of the CTP peptide with an ester linkage. 
     
     
         72 . A peptide comprising Cardiac-specific Targeting-Peptide (CTP) of the sequence of SEQ ID NO: 1, 2, 4, 5, 6, 7, 8, 9, 10, 11, 28, 29, or 34 linked to a small molecule selected from resveratrol, N-acetyl-cysteine, N-tert-butyl-α-phenylnitrone, and 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl. 
     
     
         73 . A method of treating a human subject suffering from a diastolic heart failure (DHF), comprising introducing a cargo into a cardiac muscle cell of the human subject by administering to the human subject the peptide of any one of  claims 66 - 69 , wherein the SS peptide scavenges H 2 O 2 , and ONOO—, and/or LDL oxidation. 
     
     
         74 . The method of  claim 73 , wherein the cargo is selected from the group consisting of SS-02 and SS-031. 
     
     
         75 . A method of treating aging and/or oxidative stress related disease and/or disorder in a subject in thereof, comprising introducing the peptide of any one of  claims 66 - 69  into an organ or a tissue of the subject. 
     
     
         76 . The method of  claim 75 , wherein the disease and/or disorder is selected from ischemia-reperfusion injury, neurodegenerative disease, diabetes, inflammatory diseases such as atherosclerosis, arthritis, and hepatitis. 
     
     
         77 . The peptide of any one of  claims 66 - 72  for use in a method of treating a subject by inhibiting mitochondrial permeability transition (MPT), preventing mitochondrial swelling, and/or reducing cytochrome c release in response to Ca 2+  overload. 
     
     
         78 . The peptide of any one of  claims 66 - 72  for use in a method of treating a subject by minimizing mitochondrial permeability transition (MPT)-induced ROS accumulation, thereby reducing oxidative damage on mitochondria. 
     
     
         79 . A method of treating a subject suffering from a metabolic defect that damages the heart, comprising introducing a peptide with ROS scavenging properties into a cardiac muscle cell of the subject, comprising administering, to the subject, the CTP linked to an SS peptide of any one of  claims 66 - 69 , wherein the SS peptide corrects the metabolic defect. 
     
     
         80 . The method of  claim 79 , wherein the metabolic defect is Gaucher's disease.