Chimeric alkaline phosphatase-like proteins
Inventors
Raaben, Willem • Jonk, Luigi Johannes Cornelius • Van Den Berg, Erik Jan • Van Elsas, Andrea • MILLÁN, José Luis
Assignees
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Publication Number
US-10822597-B2
Publication Date
2020-11-03
Expiration Date
Abstract
The invention relates to improved alkaline phosphatases, pharmaceutical compositions comprising improved alkaline phosphatases and the use of improved alkaline phosphatases for preventing, treating or curing diseases.
Core Innovation
The invention concerns chimeric improved alkaline phosphatase-like protein (LVL-RecAP) variants with defined sequence-identity relationships to reference ALPI/ALPP/PLAP/ALPI catalytic-region segments. The proteins are defined by sequence identity requirements to reference sequences SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7 for specified consecutive amino-acid segment lengths, together with a full-length sequence identity requirement to SEQ ID NO: 1. Specific amino acids are fixed at positions 279, 328, and 478, with leucine at position 279, valine at position 328, and leucine at position 478.
The described proteins are characterized by improved zinc-independence versus prior catALPI/crownALPP, and improved stability, including resistance under zinc chelation with EDTA. The partial content further reports altered organ pharmacokinetics with lower blood levels and higher distribution favoring skin, kidney, spleen, liver, lung, brain, fat, bone, and colon. Biodistribution and related measurements are used to support these differences relative to catALPI/crownALPP.
The partial content states therapeutic utility in treating zinc-deficiency-associated inflammatory and kidney diseases, and therapeutic claims for hypophosphatasia (HPP). It reports improvements including survival, body weight, skeletal and dentoalveolar phenotypes, and craniofacial defects, together with plasma PPi reduction and reduced renal inflammation markers in cell and animal models. The document includes gene therapy-related embodiments using polynucleotide, vectors, and host cells, and also includes pharmaceutical compositions and administration routes, supported by described preclinical and human first-in-man safety/tolerability study design elements.
Claims Coverage
The independent claim covers a phosphatase-activity protein defined by three sequence-identity constraints to SEQ ID NOs: 5, 6, and 7 over specified consecutive amino-acid lengths, a high full-length identity to SEQ ID NO: 1, and fixed amino-acid requirements at positions 279, 328, and 478. Dependent claims refine these sequence constraints and further define additional implementation elements such as vectors and pharmaceutical compositions.
Phosphatase-activity chimeric sequence-defined protein
A protein having phosphatase activity, wherein at least 200 consecutive amino acids have at least 90% sequence identity with SEQ ID NO: 5, at least 50 consecutive amino acids have at least 90% sequence identity with SEQ ID NO: 6, and at least 40 consecutive amino acids have at least 90% sequence identity with SEQ ID NO: 7, with the full length protein having at least 95% sequence identity with the full length amino acid sequence of SEQ ID NO: 1, and with the proviso that the amino acid at position 279 is leucine (L), position 328 is valine (V), and position 478 is leucine (L).
Coverage centers on the sequence-defined phosphatase-activity protein specified by identity relationships to SEQ ID NOs: 5, 6, and 7 over consecutive segments, high full-length identity to SEQ ID NO: 1, and fixed residues at positions 279, 328, and 478. Dependent claims tighten identity thresholds and/or specify additional segment length ranges while retaining the same fixed residues, and further include embodiments involving vectors, polynucleotides, and pharmaceutical compositions with pharmaceutically acceptable carriers, diluents, or excipients.
Stated Advantages
Improved zinc-independence versus prior catALPI/crownALPP.
Improved stability, including resistance under zinc chelation with EDTA.
Altered organ pharmacokinetics favoring skin, kidney, spleen, liver, lung, brain, fat, bone, and colon, together with lower blood levels.
Therapeutic efficacy in hypophosphatasia (HPP), including unexpected efficacy without a bone-targeting moiety.
Plasma PPi reduction.
Reduced renal inflammation markers in cell and animal models.
Reported improvements in survival, body weight, skeletal and dentoalveolar phenotypes, and craniofacial defects.
Documented Applications
Treating zinc-deficiency-associated inflammatory and kidney diseases.
Therapeutic use for hypophosphatasia (HPP).
Gene therapy-related embodiments using a polynucleotide encoding the protein, vectors, and host cells.
Use of pharmaceutical compositions comprising the protein with pharmaceutically acceptable carriers, diluents, or excipients, including described human first-in-man safety/tolerability study design elements.
Assessment in cell and animal models including a mouse HPP model and a rat LPS-induced AKI model, with measurements including renal inflammation markers.
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