Variant of bpifb4 protein

Abstract

The present invention relates to a variant of BPIFB4 (Bactericidal/Permeability Increasing protein family B, member 4) protein and to its use for the treatment of pathologies involving impairment of nitric oxide signalling.

1 . A BPIFB4 protein variant or fragment thereof, having an amino acid sequence with at least 85% homology to the amino acid sequence of SEQ ID NO: 1, said sequence comprising a Valine at the position corresponding to position 229 of SEQ ID NO: 1, a Threonine at the position corresponding to position 281 SEQ ID NO: 1, a Phenylalanine at a position corresponding to position 488 of SEQ ID NO: 1 and a Threonine at a position corresponding to position 494 of SEQ ID NO: 1. 
     
     
         2 . A BPIFB4 protein variant as claimed in  claim 1  having the amino acid sequence of SEQ.ID NO: 1. 
     
     
         3 . A protein having a sequence comprising the amino acid sequence of a BPIFB4 protein variant or fragment thereof according to  claim 1  linked to a sequence useful for identifying the BPIFB4 protein variant for targeting the BPIFB4 protein variant or fragment thereof to a specific organ or tissue. 
     
     
         4 . A protein as claimed in  claim 3  which is a chimeric protein. 
     
     
         5 . (canceled) 
     
     
         6 . A polynucleotide encoding a BPIFB4 protein variant or fragment thereof as claimed in  claim 1 . 
     
     
         7 . A polynucleotide encoding the protein of  claim 3 . 
     
     
         8 . A polynucleotide comprising the nucleotide sequence of SEQ ID NO: 2 or a fragment thereof which comprises the nucleotides encoding Valine 229, Threonine 281, Phenylalanine 488 and Threonine 494 of SEQ ID NO: 1. 
     
     
         9 . A vector containing a polynucleotide as claimed in  claim 6 , operatively linked to expression control sequences. 
     
     
         10 . A vector as claimed in  claim 9 , which is a viral vector. 
     
     
         11 . (canceled) 
     
     
         12 . A method for the treatment of an endothelial dysfunction due to release of NO from endothelial cells below physiological levels or to a decrease in the activity of eNOS or to another clinical condition wherein it is beneficial to obtain an increase in the activation of eNOS, the method comprising administering to a subject in need of said treatment a therapeutically effective amount of a member selected from the group consisting of:
 a BPIFB4 protein variant or fragment thereof, having an amino acid sequence with at least 85% homology to the amino acid sequence of SEQ ID NO: 1 and comprising a Valine at the position corresponding to position 229 of SEQ ID NO: 1, a Threonine at the position corresponding to position 281 SEQ ID NO: 1, a Phenylalanine at a position corresponding to position 488 of SEQ ID NO: 1 and a Threonine at a position corresponding to position 494 of SEQ ID NO: 1, said protein variant or fragment optionally linked to a sequence useful for targeting the BPIFB4 protein variant or fragment thereof to a specific organ or tissue;   a polynucleotide encoding said BPIFB4 protein variant or fragment thereof; and   a vector containing a polynucleotide as claimed in  claim 6 , operatively linked to expression control sequences.   
     
     
         13 . The method of  claim 12  wherein treatment of said endothelial dysfunction or other clinical condition results in the prevention, reduction of the risk, amelioration or treatment of a pathology selected from arterial hypertension, atherosclerosis, diabetes mellitus, dyslipidemia, renal failure, metabolic syndrome, stroke, myocardial Infarction, erectile dysfunction, neurodegenerative diseases, multiple sclerosis, cognitive disorders, retinal degeneration, uveoretinitis, vascular retinopathy, cataract, glaucoma, coronary spastic angina, thrombosis, pulmonary hypertension, pre-eclampsia, vasculitis, cancer, inflammatory disorders, venus insufficiency, genetic diseases with reduced eNOS activity and NO production, and MTHFR gene variations. 
     
     
         14 . The method of  claim 12  wherein the clinical condition is post-exercise fatigue in muscular dystrophy or a vascular occlusion wherein in the case of evascular occlusion the variant or fragment thereof is used as a co-adjuvant in the implantation of one or more stents. 
     
     
         15 . A host cell transfected with a vector as claimed in  claim 9 . 
     
     
         16 . A method of recombinantly producing a BPIFB4 protein variant or fragment thereof having an amino acid sequence with at least 85% homology to the amino acid sequence of SEQ ID NO: 1 and comprising a Valine at the position corresponding to position 229 of SEQ ID NO: 1, a Threonine at the position corresponding to position 281 SEQ ID NO: 1, a Phenylalanine at a position corresponding to position 488 of SEQ ID NO: 1 and a Threonine at a position corresponding to position 494 of SEQ ID NO: 1, said protein variant or fragment optionally linked to a sequence useful for targeting the BPIFB4 protein variant or fragment thereof to a specific organ or tissue,
 the method comprising culturing a host cell transfected with a vector containing a polynucleotide encoding said protein variant or fragment under conditions allowing expression of the protein variant or fragment and recovering said protein variant or fragment. 
 
     
     
         17 . A pharmaceutical composition comprising a BPIFB4 protein variant or fragment thereof, having an amino acid sequence with at least 85% homology to the amino acid sequence of SEQ ID NO: 1 and comprising a Valine at the position corresponding to position 229 of SEQ ID NO: 1, a Threonine at the position corresponding to position 281 SEQ ID NO: 1, a Phenylalanine at a position corresponding to position 488 of SEQ ID NO: 1 and a Threonine at a position corresponding to position 494 of SEQ ID NO: 1, said protein variant or fragment optionally linked to a sequence useful for targeting the BPIFB4 protein variant or fragment thereof to a specific organ or tissue, said variant or fragment admixed with a pharmaceutically acceptable carrier and/or excipient.