Isis Innovation Limited

Oxford Biomedica (uk) Limited

Institute of Virology of the Slovak Academy of Sciences

Bayer Healthcare LLC

Oxford University Innovation Ltd

Peter John Ratcliffe

Christopher William Pugh

Christopher Joseph Schofield

Adrian Llewllyn Harris

Patrick Henry Maxwell

Matthew Edward Cockman

Wei Ge

Kirsty Sarah Hewitson

John David Firth

Ian James Stratford

Luke McNeill

Michael Arnold McDonough

Armin Thalhammer

Christoph Loenarz

Jonathan Mark Elkins

Norma Masson

Mathew Louis Coleman

Jonathan Mark Gleadle

Alexander Wolf

A novel class of hydroxylases is described having the amino acid sequence of SEQ ID NO: 2, 4, 6 and 8, and variants and fragments thereof having HIF hydroxylation activity. The polypeptides of the invention have in particular prolyl hydroxylase activity. An assay method monitors the interaction of the HIF hydroxylase with a substrate. Modulators of HIF hydroxylase are provided for use in the treatment of a condition associated with increased or decreased HIF levels or activity or for the treatment of a condition where it is desirable to modulate HIF levels or activity.

Antibody against human HIF hydroxylase

The present invention relates to assays for monitoring activity of Mina53 and NO66 activities, in particular, to assays for identifying modulators of Mina53 and NO66 activities. The invention also relates to assays to monitor the histidinyl hydroxylase activity of Mina53 and NO66 on their substrates, the human ribosomal protein Rpl27a and Rpl8 respectively. The invention also enables the introduction of S-3-hydroxyhistidinyl residues into peptides and proteins.

Assay for histidinyl hydroxylase activity

The present invention relates to assays for monitoring activity of OGFOD1 activity, in particular, to assays for identifying modulators of OGFOD1 activity. The invention also relates to assays to monitor the prolyl hydroxylase activity of OGFOD1 on its substrate, the human ribosomal protein RPS23. The invention also enables the introduction of 3-hydroxyprolyl residues into peptides and proteins.

Method for assaying OGFOD1 activity

Assay for identifying a modulator of HIF hydroxylase

Identified herein is the location of the MN protein binding site, and MN proteins/polypeptides that compete for attachment to vertebrate cells with immobilized MN protein. Such MN proteins/polypeptides prevent cell-cell adhesion and the formation of intercellular contacts. The MN protein binding site is a therapeutic target that can be blocked by organic or inorganic molecules, preferably organic molecules, more preferably proteins/polypeptides that specifically bind to that site. Therapeutic methods for inhibiting the growth of preneoplastic/neoplastic vertebrate cells that abnormally express MN protein are disclosed. Vectors are provided that encode the variable domains of MN-specific antibodies and a flexible linker polypeptide separating those domains. Further vectors are disclosed that encode a cytotoxic protein/polypeptide operatively linked to the MN gene promoter or a MN gene promoter fragment comprising the HIF-1 consensus binding sequence, and which vectors preferably further encode a cytokine. The MN gene promoter is characterized, and the binding site for a repressor of MN transcription is disclosed. Further, the hypoxia inducibility of the MN gene and the uses of such inducibility are disclosed.

MN gene and protein

A method of identifying, screening, characterising or designing a chemical entity, which mimics or binds to FIH, is described. The method comprises comparing a structural model of FIH with a structural model for said chemical entity, wherein said structural model of FIH is derived from structural factors or structural coordinates determined by subjecting to X-ray diffraction measurements a crystal comprising FIH. Such chemical entities may be used in the treatment of a condition associated with increased or decreased HIF levels or activity.

Method of identifying a chemical entity which is a hydroxylase modulator

The invention relates to the finding that the VHL tumour suppressor protein regulates hypoxia inducible factor &#x3b1; subunits, by targeting HIF &#x3b1; for destruction in normoxic, but not hypoxic cells. The invention provides assays for modulators of this interaction, and peptides based upon HIF &#x3b1; subunit sequences which may modulate this interaction.

Interaction between the VHL tumor suppressor and hypoxia inducible factor, and assay methods relating thereto

The invention provides a compound of one of the formulae (A), (B), (C), (D), (E), (F) as herein defined, or a salt thereof, for use in the treatment of a condition associated with increased or decreased HIF levels or activity, or a condition in which an increase or decrease in HIF levels or activity may be beneficial. 

HIF hydroxylase inhibitors

A method of identifying an agent which modulates 2-oxoglutarate dependent oxygenase activity, the method comprising contacting a 2-oxoglutarate dependent oxygenase and a test agent in the presence of a substrate comprising one or more ankyrin repeat, or fragment thereof, in conditions under which the substrate is hydroxylated in the absence of the test agent; and determining hydroxylation of the substrate.

Assays for identifying modulators of the hydroxylation of ankyrin repeat proteins by 2-oxoglutarate dependent oxygenase and methods of using the same

A method of identifying an agent which modulates hydroxylation of hypoxia inducible factor (HIF), comprises contacting a HIF asparagine hydroxylase and a test substance in the presence of a substrate of the hydroxylase under conditions in which asparagine in the substrate is hydroxylated in the absence of the test substance; and determining hydroxylation of the substrate. Preferably the substrate is a HIF polypeptide comprising HIF-1&#x3b1;, a fragment thereof comprising Asn 803 of HIF-1&#x3b1; or a peptide analogue of HIF-1&#x3b1; or fragment thereof comprising an asparagine equivalent to Asn 803 of HIF-1&#x3b1; and wherein hydroxylation of Asn 803 or of a said equivalent asparagine is determined.

Assays for modulators of asparaginyl hydroxylase

The invention relates to the finding that the VHL tumour suppressor protein regulates hypoxia inducible factor &agr; subunits, by targeting HIF &agr; for destruction in normoxic, but not hypoxic cells. The invention provides assays for modulators of this interaction, and peptides based upon HIF &agr; subunit sequence which may modulate this interaction.

Nucleic acid constructs comprising hypoxia response elements in operable linkage with a coding sequence of a gene of interest, and methods for expressing a nucleic acid sequence using the constructs, are disclosed. In particular, such nucleic acid constructs comprise genes encoding prodrug activation systems or cytokines.

Growing community of inventors

Oxford, United Kingdom

Peter John Ratcliffe