Materials and methods for treating disorders associated with sulfatase enzymes

Abstract

The subject invention concerns materials and methods for treating or preventing disease and conditions associated with various sulfatase enzymes that are defective or that are not properly expressed in a person or animal. In one embodiment, the disease is Sanfilippo A (MPS-IIIA) disease. The subject invention also concerns materials and methods for treating or preventing multiple sulfatase deficiency (MSD) in a person or animal. Compounds of the invention include a fusion protein comprising i) a mammalian sulfatase, or an enzymatically active fragment or variant thereof, and ii) a plant lectin or a binding subunit thereof. In a specific embodiment, the mammalian sulfatase is a human sulfatase, or an enzymatically active fragment or variant thereof. Polynucleotides encoding the fusion proteins are also contemplated for the subject invention. The subject invention also concerns materials and methods for producing proteins of the invention.

1 . A fusion protein comprising i) a mammalian sulfatase, or an enzymatically active fragment or variant thereof, and ii) a plant lectin or a binding subunit thereof, or a fusion protein comprising i) a mammalian sulfatase modifying factor 1 (SUMF1), or an enzymatically active fragment or variant thereof, and ii) a plant lectin or a binding subunit thereof; or a polynucleotide encoding said fusion protein; or
 a human SUMF1 protein expressed in plant cells comprising transforming a plant cell with an expression vector comprising a nucleotide sequence for translational expression of the SUMF1 enzymatically active fragment or variant thereof, produced in a plant or plant cell.   
     
     
         2 . The fusion protein according to  claim 1 , wherein the mammalian sulfatase is N-acetylgalactosamine-6-sulfatase, N-acetylglucosamine-6-sulfatase, N-sulphoglucosamine sulphohydrolase, sulfamidase, extracellular sulfatase Sulf-1 (hSulf1), extracellular sulfatase Sulf-2 (hSulf2), iduronate 2-sulfatase, arylsulfatase A (ASA), arylsulfatase B (ASB), steryl-sulfatase, arylsulfatase D (ASD), arylsulfatase E (ASE), arylsulfatase F (ASF), arylsulfatase G (ASG), arylsulfatase H (ASH), arylsulfatase I (ASI), arylsulfatase J (ASJ), or arylsulfatase K (ASK). 
     
     
         3 . The fusion protein according to  claim 1 , wherein the mammalian sulfatase comprises the amino acid sequence of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, or 34, or an enzymatically active fragment or variant thereof. 
     
     
         4 . The fusion protein according to  claim 1 , wherein the plant lectin is a lectin from Table 2 or Table 3 of the specification. 
     
     
         5 . The fusion protein according to  claim 1 , wherein the plant lectin is the non-toxic subunit of ricin (RTB) or nigrin (NBB). 
     
     
         6 . The fusion protein according to  claim 1 , wherein the fusion protein comprises an endoplasmic reticulum (ER) retention sequence. 
     
     
         7 . The fusion protein according to  claim 6 , wherein the ER retention sequence comprises KDEL. 
     
     
         8 . The fusion protein according to  claim 1 , wherein the mammalian sulfatase is linked to the plant lectin by a linker sequence of amino acids. 
     
     
         9 - 10 . (canceled) 
     
     
         11 . A method for treating or preventing a disease or condition associated with a sulfatase enzyme or a sulfatase modifying factor 1 (SUMF1) protein in a person or animal, comprising administering to the person or animal a therapeutically effective amount of a SUMF1 protein or a fusion protein of  claim 1 . 
     
     
         12 . The method according to  claim 11 , wherein the disease or condition is mucoposysaccharidosis IVA (MPS-IVA), Morquio A syndrome, mucoposysaccharidosis IIID (MPS-IIID), Sanfilippo D syndrome, mucopolysaccharidosis IIIA (MPS-IIIA), Sanfilippo A syndrome, mucopolysaccharidosis II (MPS-II), Hunter syndrome, metachromatic leukodystrophy (MLD), mucopolysaccharidosis VI (MPS-VI), Maroteaux-Lamy syndrome, X-linked ichthyosis (XLI), or chondrodysplasia punctata 1 (CDPX1). 
     
     
         13 . The method according to  claim 12 , wherein the fusion protein is administered by intravenous infusion or injection, or by inhalation via nasal cavity or lung, or orally, ocularly, vaginally, anally, rectally, or transmembraneously or transdermally, subcutaneously, intradermally, intravenously, intramuscularly, intraperitoneally, or intrasternally, such as by injection. 
     
     
         14 . (canceled) 
     
     
         15 . The fusion protein according to  claim 1 , wherein the SUMF1 comprises the amino acid sequence of SEQ ID NO:36, or an enzymatically active fragment or variant thereof. 
     
     
         16 - 25 . (canceled) 
     
     
         26 . A method for producing a sulfatase fusion protein and/or a mammalian SUMF1 protein and/or a SUMF1 fusion protein of  claim 1 , and/or a mammalian sulfatase, or an enzymatically active fragment or variant of any of the proteins, comprising expressing in a plant or plant cell a polynucleotide encoding a mammalian sulfatase fusion protein and/or a polynucleotide encoding a sulfatase modifying factor 1 (SUMF1) protein or a SUMF1 fusion protein, and/or a polynucleotide encoding a mammalian sulfatase, or an enzymatically active fragment or variant of any of the proteins. 
     
     
         27 - 28 . (canceled) 
     
     
         29 . The method according to  claim 26 , wherein the SUMF1 protein or the SUMF1 fusion protein comprises an ER retention signal. 
     
     
         30 . The method according to  claim 29 , wherein the ER retention signal comprises KDEL sequence. 
     
     
         31 - 38 . (canceled) 
     
     
         39 . The method according to  claim 26 , wherein the plant or plant cell is transiently or stably transformed with one or both of the polynucleotides. 
     
     
         40 . (canceled) 
     
     
         41 . The human SUMF1 protein of  claim 1 , wherein the SUMF1 protein shows enzymatic and biological activity with capacity to activate human sulfatases expressed in plant cells. 
     
     
         42 . (canceled) 
     
     
         43 . The human SUMF1 protein of  claim 41 , wherein the SUMF1 protein activates a sulfatase in plant cells by catalyzing the conversion of a relevant cysteine to a FGly residue required for activating enzymatic activity of the sulfatase. 
     
     
         44 - 66 . (canceled) 
     
     
         67 . The fusion protein according to  claim 1 , wherein the fusion protein comprises the amino acid sequence of any of SEQ ID NOs:39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, or 69, or an enzymatically active fragment or variant thereof; or wherein the fusion protein comprises the amino acid sequence of any of SEQ ID NOs:71, 73. 75, 77, 79, or 81, or an enzymatically active fragment or variant thereof. 
     
     
         68 - 93 . (canceled)