METHODS AND PHARMACEUTICAL COMPOSITIONS FOR THE TREATMENT OF ISCHEMIC CONDITIONS

Abstract

The present invention relates to methods and pharmaceutical compositions for the treatment of ischemic conditions. In particular, the present invention relates to a method of treating an ischemic condition in a subject in need thereof comprising administering the subject with a polypeptide comprising an amino acid sequence having at least 70% of identity with the amino acid sequence ranging from the amino acid residue at position 186 to the amino acid residue at position 406 in SEQ ID NO: 1.

Claims

1. A method of treating an ischemic condition in a subject in need thereof comprising administering to the subject a polypeptide comprising an amino acid sequence having at least 70% of identity with an amino acid sequence ranging from an amino acid residue at position 186 to an amino acid residue at position 406 in SEQ ID NO:1, provided that said polypeptide is not SEQ ID NO:1

2. The method of claim 1 wherein the ischemic condition is selected from the group consisting of renal ischemia, retinal ischemia, brain ischemia and myocardial ischemia.

3. The method of claim 1 wherein the ischemic condition is selected from the group consisting of coronary artery bypass graft surgery, global cerebral ischemia due to cardiac arrest, focal cerebral infarction, cerebral hemorrhage, hemorrhage infarction, hypertensive hemorrhage, hemorrhage due to rupture of intracranial vascular abnormalities, subarachnoid hemorrhage due to rupture of intracranial arterial aneurysms, hypertensive encephalopathy, carotid stenosis or occlusion leading to cerebral ischemia, cardiogenic thromboembolism, stroke, spinal stroke and spinal cord injury, vasculitis, macular degeneration, myocardial infarction, cardiac ischemia and superaventicular tachyarrhytmia.

4. The method of claim 1 wherein the polypeptide is administered sequentially or concomitantly with a standard method selected from the group consisting of reperfusion of the ischemic organ by angioplasty, percutaneous coronary intervention, thrombolysis and coronary surgery.

5. The method of claim 1 wherein the polypeptide is administered sequentially or concomitantly with a thrombolyic agent selected from the group consisting of reteplase, alteplase, urokinase, prourokinase, anisoylated purified streptokinase activator complex, and streptokinase.

6. The method of claim 1 wherein the polypeptide has an amino acid sequence having 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80; 81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 98; or 99, or 100% of identity with the amino acid sequence from the amino acid residue at position 186 to the amino acid residue at position 406 in SEQ ID NO:1

7. The method of claim 1 wherein the polypeptide comprises an amino acid sequence having at least 70% identity with an amino acid sequence ranging from the amino acid residue at position 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; 32; 33; 34; 35; 36; 37; 38; 39; 40; 41; 42; 43; 44; 45; 46; 47; 48; 49; 50; 51; 52; 53; 54; 55; 56; 57; 58; 59; 60; 61; 62; 63; 64; 65; 66; 67; 68; 69; 70; 71; 72; 73; 74; 75; 76; 77; 78; 79; 80; 81; 82; 83; 84; 85; 86; 87; 88; 89; 90; 91; 92; 93; 94; 95; 96; 97; 98; 99; 100; 101; 102; 103; 104; 105; 106; 107; 108; 109; 110; 111; 112; 113; 114; 115; 116; 117; 118; 119; 120; 121; 122; 123; 124; 125; 126; 127; 128; 129; 130; 131; 132; 133; 134; 135; 136; 137; 138; 139; 140; 141; 142; 143; 144; 145; 146; 147; 148; 149; 150; 151; 152; 153; 154; 155; 156; 157; 158; 159; 160; 161; 162; 163; 164; 165; 166; 167; 168; 169; 170; 171; 172; 173; 174; 175; 176; 177; 178; 179; 180; 181; 182; 183; 184; or 185 to the amino acid residue at position 406 in SEQ ID NO:1.

8. The method of claim 1 wherein the polypeptide is fused a Fc domain of an immunoglobulin.

9. A method of treating acute myocardial infarction in a patient in need thereof comprising the steps of i) restoring blood supply in the cardiac ischemic tissue, and ii) preserving vascular endothelial cell barrier integrity of said cardiac ischemic tissue by administering to said patient a therapeutically effective amount of an ANGPTL4 polypeptide to reduce infarct size, hemorrhage, and no-reflow, where steps i) and ii) are performed sequentially or concomitantly and wherein the ANGPTL4 polypeptide comprises an amino acid sequence having at least 70% identity with an amino acid sequence ranging from the amino acid residue at position 186 to the amino acid residue at position 406 in SEQ ID NO:1, provided that said polypeptide is not SEQ ID NO:1.

10. A method of treating an ischemic condition in a subject in need thereof comprising administering to the subject a nucleic acid molecule encoding a polypeptide as defined in claim 1.

Description

FIGURES

[0023] FIG. 1 shows the amino acid sequence of ANGPTL4, the fibrinogen-like domain; (FLD) corresponds to the sequence in bold and underlined in the whole sequence of ANGPTL4.

[0024] FIG. 2 shows the comparison between the coiled-coil domain (CCD) polypeptide and the fibrinogen-like domain (FLD) polypeptide in the reduction of infarct size

[0025] FIG. 3 shows that the ANGPTL4 FLD polypeptide administration protocol (bolus+perfusion) leads to a significant reduction of the infarct size in mice that received the polypeptide compared to controls A. ANGPTL4 FLD polypeptide (bolus+various duration of infusion after reperfusion) in mice. B. ANGPTL4 polypeptide (bolus+2 h infusion after reperfusion with 2 doses) in mice.

[0026] FIG. 4 shows that administration of ANGPTL4 FLD polypeptide (bolus+perfusion) leads to a significant reduction of the infarct size in pigs.

EXAMPLE 1: COMPARISON BETWEEN THE CCD POLYPEPTIDE AND THE FLD POLYPEPTIDE OF ANGPTL4

[0027] Mice were anesthetized by an intraperitoneal injection of sodium pentobarbital. Coronary artery occlusion (CAO) was induced during 30-min. One min before reperfusion, the animals received a bolus of the human recombinant CC (coiled-coil) or FLD (fibrinogen-like domain) polypeptide of ANGPTL4 (or vehicle for the control animals). Just after reperfusion (i.e. release of the occlusion) the animals were infused with an amount of the same recombinant polypeptide (or vehicle for the control animals) for 30 min. The infarct area was then identified by Evans blue staining at 24 h after ischemia when the animals were euthanized. The results are depicted in FIG. 2 and clearly show that the coiled-coil domain (CC) does not confer protection in contrast to the fibrinogen-like domain (FLD) domain. In conclusion, the therapeutic effects observed with ANGPTL4 in ischemic conditions are not due to the coiled-coil domain as suggested in the prior at (WO2011089152) but are brought by the fibrinogen-like domain.

EXAMPLE 2: MOUSE MODEL OF MYOCARDIAL INFARCTION

[0028] Mice were anesthetized by an intraperitoneal injection of sodium pentobarbital. Coronary artery occlusion (CAO) was induced during 30-min. One min before reperfusion, the animals received a bolus of the human recombinant FLD (fibrinogen-like domain) polypeptide of ANGPTL4 (or vehicle for the control animals). Just after reperfusion (i.e. release of the occlusion) the animals were infused with an amount of the same recombinant FLD polypeptide (or vehicle for the control animals) for 30 min, 1 hour and 2 hours. The infarct area was then identified by Evans blue staining at 24 h after ischemia when the animals were euthanized. The results are depicted in FIG. 3 and clearly show that this protocol of administration lead to a significant reduction of the infarct size in the animals which received the polypeptide.

EXAMPLE 3: PIG MODEL OF MYOCARDIAL INFARCTION

[0029] Coronary artery occlusion (CAO) was induced during 30-min. One min before reperfusion, the animals received a bolus of human recombinant FLD (fibrinogen-like domain) polypeptide of ANGPTL4 (or vehicle for the control animals). Just after reperfusion (i.e. release of the occlusion) the animals were infused with an amount of the same FLD polypeptide (or vehicle for the control animals) for 2 hours. The infarct area was then identified by Evans blue staining at 24 h after ischemia when the animals were euthanized. The results are depicted in FIG. 4 and clearly show that this protocol of administration lead to a significant reduction of the infarct size in the animals which received the polypeptide.

REFERENCES

[0030] Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.