Polypeptides, antibody variable domains & antagonists

Abstract

The present disclosure relates to immunoglobulin single variable domains (dAbs) e.g., dAbs which are protease resistant, and also to formulations, and compositions comprising such dAbs for ocular delivery and to their uses to treat ocular diseases and conditions.

1 . A composition comprising an immunoglobulin single variable domain that binds to a target molecule, for ocular delivery. 
     
     
         2 . The composition according to  claim 1 , wherein said immunoglobulin single variable domain is resistant to a protease, wherein the protease is selected from the group consisting of: ocular proteases, caspases, calpains, matrix metalloproteases, disintegrins, metalloproteinases, ADAMs, ADAM with motifs, proteosomes, tissue plasminogen activator, secretases, cathepsin B, cathepsin D, cystatin C, serine protease PRSS1, and ubiquitin proteosome pathway. 
     
     
         3 . A composition according to  claim 1 , which comprises an immunoglobulin single variable domain which binds to a target molecule selected from the group consisting of: VEGF, TNFalpha, TNFalphaR, IL-1, IL-1r, TNFalphaR1, TGFbeta, IL-6, IL-8, IL-17, IL-21, IL-23, CD20, Nogo-a, myelin associated glycoprotein and beta amyloid. 
     
     
         4 . The composition according to  claim 3 , wherein said immunoglobulin single variable domain which binds to VEGF comprises an amino acid sequence that is at least 97% identical to an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of DOM15-26-593 shown in SEQ ID NO 1 and (b) the amino acid sequence of DOM15-26-593-Fc shown in SEQ ID NO 2. 
     
     
         5 . The composition according to  claim 4 , wherein the immunoglobulin single variable domain comprises valine at position 6, wherein numbering is according to Kabat. 
     
     
         6 . The composition according to  claim 4 , wherein the immunoglobulin single variable domain comprises leucine at position 99, wherein numbering is according to Kabat. 
     
     
         7 . The composition according to  claim 4 , wherein the immunoglobulin single variable domain comprises lysine at position 30, wherein numbering is according to Kabat. 
     
     
         8 . The composition according to  claim 4 , wherein the immunoglobulin single variable domain comprises an amino acid sequence that is identical to an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of DOM15-26-593 shown in SEQ ID NO 1 and (b) the amino acid sequence of DOM15-26-593 Fc shown in SEQ ID NO 2. 
     
     
         9 . The composition according to  claim 3 , wherein said immunoglobulin single variable domain which binds to IL-1 comprises an amino acid sequence that is at least 97% identical to an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of DOM 4-130-54 shown in SEQ ID NO 5 and (b) the amino acid sequence of DOM 0400 PEG shown in SEQ ID NO 4. 
     
     
         10 . The composition according to  claim 9 , wherein said immunoglobulin single variable domain which binds to IL-1 comprises an amino acid sequence that is identical to an amino acid sequence selected from the group consisting of: (a) the amino acid sequence of DOM 4-130-54 shown in SEQ ID NO 5 and (b) the amino acid sequence of DOM 0400 PEG shown in SEQ ID NO 4. 
     
     
         11 . The composition according to  claim 3 , wherein said immunoglobulin single variable domain which binds to TNFalphaR1 and comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of Dom 1h-131-206 shown in SEQ ID NO 6. 
     
     
         12 . The composition according to  claim 3 , wherein said immunoglobulin single variable domain which binds to TNFalphaR1 and comprises an amino acid sequence that is identical the amino acid sequence of Dom 1 h-131-206 shown in SEQ ID NO 6. 
     
     
         13 . A composition according to  claim 1  which further comprises a domain of an antibody constant region, wherein the antibody constant region is an antibody Fc region. 
     
     
         14 . The composition according to  claim 9 , wherein said antibody Fc region has the amino acid. Fc sequence shown in SEQ ID NO 3. 
     
     
         15 . The composition according to  claim 9 , wherein the immunoglobulin single variable domain is present as a fusion with an Fc and has an amino acid sequence identical to the amino acid sequence of the DOM15-26-593-Fc fusion shown in SEQ ID NO 2. 
     
     
         16 . A composition comprising a naked immunoglobulin single variable domain which binds to a target molecule for delivery to one or more of the ocular regions selected from the group consisting of: the vitreous humour, the aqueous humour, the retina and the choroid. 
     
     
         17 . The composition according to  claim 16  wherein the target molecule is selected from the group consisting of: VEGF, a VEGF antagonist, TNFalpha receptor, TNFalpha receptor, IL-1, TNFalphaR1, IL-6, IL-8, IL-17, IL-21, IL-23, Nogo-a, myelin associated glycoprotein and beta amyloid. 
     
     
         18 . A composition according, to  claim 16 , wherein said immunoglobulin single variable domain is resistant to a protease, wherein the protease is selected from the group consisting of: ocular proteases, caspases, calpains, matrix metalloproteases, disintegrins, metalloproteinases, ADAMs, ADAM with thrombospondin motifs, proteosomes, tissue plasminogen activator, secretases, cathepsin B, cathepsin D, cystatin C, serine protease PRSS1, and ubiquitin proteosome pathway. 
     
     
         19 . A composition according to  claim 16 , wherein said immunoglobulin single variable domain is selected from the group consisting of: (a) an immunoglobulin single variable domain which binds to VEGF and which comprises an amino acid sequence that is at least 97% identical to an amino acid selected from the group consisting of: (i) the amino acid sequence of DOM15-26-593 or shown in SEQ ID NO 1 and (ii) the amino acid sequence of DOM15-26-593-Fc shown in SEQ ID NO 2; (b) an immunoglobulin single variable domain which binds to IL-1 and comprises an amino acid sequence that is at least 97% identical to an amino acid sequence selected from the group consisting of: (i) the amino acid sequence of DOM 4-130-54 shown in SEQ ID NO 5 and (ii) the amino acid sequence of DOM 0400 PEG shown in SEQ ID NO 4; and (c) are immunoglobulin single variable domain which binds to TNFalphaR1 and comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of Dom 1h-131-206 shown in SEQ ID NO 6. 
     
     
         20 . A composition comprising a formatted immunoglobulin single variable domain which binds to a target molecule for delivery to one or more of the ocular regions selected, from the group consisting of: the retina, the choroid and the lachrymal fluid. 
     
     
         21 . The composition according to  claim 20 , wherein the target molecule is selected from the group consisting of: VEGF, a VEGF antagonist, TNFalpha, TNFalpha receptor, IL-1, TNFalphaR1, IL-17, IL-21, IL-23, Nogo-a, myelin associated glycoprotein and beta amyloid. 
     
     
         22 . The composition according to  claim 20 , wherein said immunoglobulin single variable domain is resistant to a protease, wherein the protease is selected from the group consisting of: ocular proteases, caspases, calpains, matrix metalloproteases, disintegrins, metalloproteinases, ADAMs, ADAM with thrombospondin motifs, proteosomes, tissue plasminogen activator, secretases, cathepsin B, cathepsin D, cystatin C, serine protease PRSS1, and ubiquitin proteosome pathway. 
     
     
         23 . The composition according to  claim 16 , wherein said immunoglobulin single variable domain is selected from the group consisting of: (a) an immunoglobulin single variable domain which binds to VEGF and which comprises an amino acid sequence that is at least 97% identical to an amino acid sequence selected from the group consisting of: (i) the amino acid sequence of DOM 5-26-593 shown in SEQ ID NO 1 and (ii) the amino acid sequence of DOM 15-26-593 Fc shown in SEQ ID NO 2; (b) an immunoglobulin single, variable domain which binds to IL-1 and comprises an amino acid sequence that is at least 97% identical to an amino acid sequence selected from the group consisting of: (i) the amino acid sequence of DOM 4-130-54 shown in SEQ ID NO 5 and (ii) the amino acid sequence of DOM 0400 PEG shown in SEQ ID NO 4, and (c) an immunoglobulin single variable domain which binds to TNFalphaR1 comprises an amino acid sequence that is at least 97% identical to the amino acid sequence of Dom 1h-131-206 shown in SEQ ID NO 6. 
     
     
         24 . A composition according to  claim 20 , wherein the immunoglobulin single variable domain has a molecular weight of around 50 KDa. 
     
     
         25 . A composition according to  claim 20 , wherein the immunoglobulin single variable domain is formatted by pegylation or fusion to an antibody Fc. 
     
     
         26 . A composition according to  claim 1 , which further comprises one or more enhancers selected from the group consisting of: an ocular penetration enhancer and a viscosity enhancer. 
     
     
         27 . A composition according to  claim 1 , which further comprises a component selected from the group consisting of: a pharmaceutically acceptable carrier, a physiologically acceptable carrier, a diluent, and an excipient. 
     
     
         28 . A method of delivering a composition according to  claim 1 , directly to the eye, which comprises administering said composition to the eye by a method for topical delivery to selected from the group consisting of: eye drops, intra-ocular injection, peri-ocular administration, and a slow release formulation. 
     
     
         29 . A method for treating an eye condition which comprises administering a composition according to  claim 1  directly to the eye by a method selected from the group consisting of: intra-ocular injection, intra-vitreal injection, eye drops, peri-ocular administration, and a slow release formulation. 
     
     
         30 . A method according to  claim 28  or  29 , wherein the composition is administered to a one or more regions of the eye selected from the group consisting of: a surface of the eye, tear ducts, lachrymal glands, an infra-ocular region, anterior chamber, posterior chamber and the vitreous humour. 
     
     
         31 . A method of delivering a composition according to  claim 26 , to at least one region of the eye selected from the group consisting of: the vitreous humour, the aqueous humour, the retina, and the choroid; which comprises administering said composition to the eye by topical delivery. 
     
     
         32 . A method of delivering a composition according to  claim 20  to at least one region of the eye selected from the group consisting of: the retina, the choroid, and the lachrymal fluid; which comprises administering said composition to the eye by topical delivery. 
     
     
         33 . A process for producing a pharmaceutical composition comprising: (a) mixing a composition according to  claim 1  with (b) a pharmaceutically acceptable carrier, diluent or excipient. 
     
     
         34 . A process according to  claim 33 , wherein said pharmaceutical composition is for treating an eye condition or disease. 
     
     
         35 . A process according to  claim 34 , wherein said eye condition or disease is selected from the group consisting of: age-related macular degeneration, uveitis, glaucoma, dry eye, diabetic retinopathy, and diabetic macular oedema.