Urate oxidase-albumin conjugate, preparation method thereof, and use thereof
The present specification discloses a urate oxidase-albumin conjugate, a preparation method thereof, a urate oxidase variant contained in the urate oxidase-albumin conjugate, and a preparation method thereof. The urate oxidase-albumin conjugate is characterized in that three or more albumins are conjugated to the urate oxidase variant through a linker, thereby improving half-life and reducing immunogenicity. In addition, the urate oxidase-albumin conjugate can be used to prevent or treat various diseases, disorders and/or indications caused by uric acid.
1. A method for treating a uric acid-related disease, comprising:
administering a therapeutically effective amount of a urate oxidase-albumin conjugate represented by [formula 1] to a subject,
Uox-[J 1 -A-J 2 -HSA] n , [formula 1]
wherein Uox is a urate oxidase variant, J 1 is a urate oxidase-linker junction, A is an anchor, J 2 is an albumin-linker junction, and HSA is Human Serum Albumin;
wherein n is 3 or 4;
the urate oxidase variant is a tetramer which is formed by four urate oxidase variant subunits that are oligomerized,
each of the urate oxidase variant subunit is represented by
(SEQ ID NO: 29)
SAVKAARYGKDNVRVYKVHKDEKTGVQTVYEMTVCV
LLEGEIETSYTKADNSVIVATDSIKNTIYITAKQN
PVTPPELFGSILGTHFIEKYNHIHAAHVNIVCHRW
TRMDIDGKPHPHSFIRDSEEKRNVQVDVVEGKGID
IKSSLSGLTVLKSTNSQFWGFLRDEYTTLKETXDR
ILSTDVDATWQWKNFSGLQEVRSHYPKFDATWATA
REVTLKTFAEDNSASVQATMYKMAEQILARQQLIE
TVEYSLPNKHYFEIDLSWHKGLQNTGKNAEVFAPQ
SDPNGLIKCTVGRSSLKSKL,
wherein the X of the SEQ ID NO: 29 is 4-(1,2,3,4-tetrazine-3-yl)-phenylalanine (frTet);
wherein the urate oxidase-linker junction is formed by Inverse Electron Demand Diels-Alder (IEDDA) reaction between a tetrazine residue of the nonnatural amino acid of the urate oxidase variant and trans-cyclooctene moiety linked to the anchor;
wherein the urate oxidase-linker junction is represented by
in which the (1) is linked to the residue of the nonnatural amino acid residue and the (2) is linked to the anchor;
wherein the albumin-linker junction is formed by a reaction between a thiol moiety of the albumin and a thiol reactive moiety of the anchor; and
wherein the albumin-linker junction is represented by
in which the (1) is linked to the albumin and the (2) is linked to the anchor.
2. The method of claim 1 , wherein the anchor is selected from the following:
wherein J 1 is a urate oxidase-linker junction, and J2 a is albumin-linker junction.
3. The method of claim 1 , wherein the uric acid-related disease is selected from hyperuricemia, acute gouty arthritis, intermittent gout, chronic nodular gout, Chronic Kidney Disease, and Tumor Lysis Syndrome (TLS).