Biological materials and uses thereof

Abstract

The invention provides compounds comprising a therapeutic agent coupled to a carrier molecule, with a minimum coupling ratio of 5:1; wherein the carrier molecule is (i) an antibody fragment or derivative thereof or (ii) an antibody mimetic or derivative thereof; and wherein the therapeutic agents are coupled onto a lysine amino acid residue; and further wherein the therapeutic agent is not a photosensitising agent. There is also provided uses, methods relating to such compounds, as well as processes for their manufacture.

1 . A compound comprising a therapeutic agent coupled to a carrier molecule, with a minimum coupling ratio of 5:1; wherein the carrier molecule is (i) an antibody fragment or derivative thereof or (ii) an antibody mimetic or derivative thereof; and wherein the therapeutic agents are coupled onto a lysine amino acid residue; and further wherein the therapeutic agent is not a photosensitising agent. 
     
     
         2 . The compound of  claim 1  wherein the functional and physical properties of the therapeutic agent and the carrier molecule are qualitatively substantially unaltered in the coupled form in comparison to the properties when in an uncoupled form. 
     
     
         3 . A compound according to  claim 1  wherein the compound has
 (a) an IC50 of 100 nM or lower; and/or 
 (b) an IC50 of at least 10-fold lower than the therapeutic agent when unconjugated; and or 
 (c) a serum half-life of at least 2 hours, optionally the serum half-life of at least 2 hours is measured in mice or in humans; and/or 
 (d) a serum half-life of at least 50% of that of the free antibody when unconjugated; and/or 
 (e) a solubility of at least 1 mg/ml in phosphate-buffered saline at room temperature; and or 
 (f) a solubility of at least 1 mg/ml in phosphate-buffered saline at room temperature in the presence of an excipient a concentration and type approved by the FDA, wherein the excipient is up to 0.5% polysorbate, 1% glycerol, 0.5% glycine, 0.1% histidine, 0.5% chlorobutanol, 5% propylene glycol, 2% benzyl alcohol, 0.05% octanoic acid and/or 0.1% N-acetyl tryptophan; and/or 
 (g) an aggregation level of up to 5% in phosphate-buffered saline at room temperature; and/or 
 
     
     
         4 .- 10 . (canceled) 
     
     
         11 . A compound according to  claim 1 , wherein the therapeutic agents, when coupled to the carrier molecule, are (h) separated by a distance of at least two amino acids (3.5 to 7.5 angstroms), and/or (i) separated by a distance of two amino acids (3.5 to 7.5 angstroms), three amino acids (9 to 12 angstroms), four amino acids (10 to 15 angstroms), five amino acids (15 to 20 angstroms) or six amino acids (20 to 25 angstroms). 
     
     
         12 . (canceled) 
     
     
         13 . A compound according to  claim 1 , wherein the therapeutic agents are directly coupled to the carrier molecule at the amino acid, optionally wherein the direct coupling to the amino acid is via an N-hydroxy-succinamide ester. 
     
     
         14 . (canceled) 
     
     
         15 . A compound according to  claim 1  wherein the therapeutic agents are indirectly coupled to the carrier molecule at the amino acid, optionally wherein the indirect coupling to the amino acid is via a thiol or maleimide. 
     
     
         16 . (canceled) 
     
     
         17 . The compound of  claim 1 , wherein the carrier molecule binds selectively to a target, optionally wherein the target is a target cell or an extracellular target molecule. 
     
     
         18 . (canceled) 
     
     
         19 . The compound according to  claim 17  wherein the carrier molecule, on binding a target cell, is internalised into the cell and following binding of the target, is decoupled from the therapeutic agent. 
     
     
         20 .- 21 . (canceled) 
     
     
         22 . The compound of  claim 1  wherein the carrier molecule is an antibody fragment that does not include the CH2 and CH3 antibody regions of a whole antibody. 
     
     
         23 . The compound of  claim 1  wherein the carrier molecule is an antibody fragment selected from scFv, Fv, Fab, F(ab′)2, Fab-SH, dsFv, be-scFv, sdAb, di-scFvs (also known as bi-scFvs), Fcabs, domain antibodies, nanobodies, VHH domains, bispecific formats such as bispecific T-cell engagers, diabodies, and tandabs. 
     
     
         24 . The compound of  claim 1  wherein the carrier molecule is an antibody mimetic selected from DARPins, affibodies, affitins, anticalins, avimers, kunitz domain peptides, adnectins, centyrins, Fynomers, IgNARs and monobodies. 
     
     
         25 . The compound of  claim 1  wherein the carrier molecule is humanised or human. 
     
     
         26 . The compound of  claim 1  wherein the carrier molecule binds specifically to HER2, EGFR, HER3, MUC1, EpCAM, CEA, Fibronectin-EDB, CD19, CD20, CD22, LeY, CD30, CD33, CD79b, GPNMB, PSMA, CD56, CD37, Folate receptor, CA6, CD27L, MUC16, CD66e, CD74, Trop-2 or guanylate cyclase. 
     
     
         27 . The compound of  claim 1  wherein the therapeutic agent is a cytotoxic agent or a cytostatic agent. 
     
     
         28 . The compound of  claim 1  wherein the therapeutic agent is selected from cell cycle progression inhibitors, angiogenesis inhibitors, MAPK signaling pathway inhibitors, PI3K/m-TOR/AKT pathway inhibitors, kinase inhibitors, HDAC inhibitors, protein chaperone inhibitors, PARP inhibitors, Wnt/Hedgehog/Notch signalling pathway inhibitors, RNA polymerase inhibitors. DNA-binding drugs, DNA damaging drugs, DNA alkylating drugs, microtubule stabilizing agents, microtubule destabilizing agents, platinum compounds, kinase inhibitors, pyridocarbazole and its derivatives, and topoisomerase I inhibitors, cemadotin, P5, P5-C5, doxorubicin, ellipticine, MMAE, paclitaxel, auristatins, maytansines, dolostatins, camptothecin, SN-38 and pyrrolobenzodiazepine dimers (PBDs), PNU-159862, indolino-benzodiazepine dimers (IGNs) and MMAF. 
     
     
         29 . (canceled) 
     
     
         30 . The compound according to  claim 1  wherein the carrier molecule is an scFv and the therapeutic agent is selected from the group consisting of cemadotin, doxorubicin, ellipticine, MMAE, P5-C5, maytansine, pyrrolobenzodiazepine dimer (PBD), and MMAF. 
     
     
         31 - 37 . (canceled) 
     
     
         38 . The compound of  claim 30  wherein the scFv binds specifically to HER2, optionally wherein the scFv has the amino acid sequence of SEQ ID NO. 2, SEQ ID NO. 4 or SEQ ID NO. 5. 
     
     
         39 . (canceled) 
     
     
         40 . A pharmaceutical composition comprising the compound of  claim 1  and a pharmaceutically-acceptable carrier, excipient or diluent. 
     
     
         41 . A compound as defined in  claim 1  for use in the diagnosis, treatment and/or prevention of disease. 
     
     
         42 .- 44 . (canceled) 
     
     
         45 . A process of making a compound as defined in  claim 1  comprising the steps of:
 providing a therapeutic agent; 
 providing a carrier molecule; 
 conjugating the therapeutic agent and the carrier molecule in the presence of at least one polar aprotic solvent and an aqueous buffer. 
 
     
     
         46 .- 54 . (canceled)