Polymer factor ix moiety conjugates

Abstract

Conjugates of a Factor IX moiety and one or more water-soluble polymers are provided. Typically, the water-soluble polymer is poly(ethylene glycol) or a derivative thereof. Also provided (among other things) are compositions comprising the conjugates, methods of making the conjugates, and methods of administering to a patient compositions comprising the conjugates.

What is claimed is: 
     
         1 . A conjugate comprising a Factor IX moiety covalently attached, either directly or through a spacer moiety comprised of one or more atoms, to a water-soluble polymer, wherein the molecular weight of the water-soluble polymer is greater than 5,000 Daltons and less than about 150,000 Daltons. 
     
     
         2 . A conjugate comprising a Factor IX moiety covalently attached at an amino acid residue, either directly or through a spacer moiety comprised of one or more atoms, to a water-soluble polymer, wherein the amino acid residue is not attached, either directly or though the spacer moiety, via a —CH 2 —C(O)—O—, —N(H)—C(O)CH 2 —O—, —C(O)—N(H)—, —N(H)—C(O)—CH 2 —O—, —C(O)—CH 2 —O—, —C(O)—CH 2 —CH 2 —C(O)—O—, diazo, or triazine linkage. 
     
     
         3 . A conjugate comprising a Factor IX moiety covalently attached, either directly or through a spacer moiety comprising of one or more atoms, to a non-linear water-soluble polymer. 
     
     
         4 . The conjugate of  claim 1 ,  2  or  3 , wherein the water-soluble polymer in each conjugate is selected from the group consisting of a poly(alkylene oxide), poly(vinyl pyrrolidone), poly(vinyl alcohol), polyoxazoline, and poly(acryloylmorpholine). 
     
     
         5 . The conjugate of  claim 4 , wherein each water-soluble polymer is a poly(alkylene oxide). 
     
     
         6 . The conjugate of  claim 5 , wherein each poly(alkylene oxide) is a poly(ethylene glycol). 
     
     
         7 . The conjugate of  claim 6 , wherein the poly(ethylene glycol) is terminally capped with an end-capping moiety selected from the group consisting hydroxy, alkoxy, substituted alkoxy, alkenoxy, substituted alkenoxy, alkynoxy, substituted alkynoxy, aryloxy and substituted aryloxy. 
     
     
         8 . The conjugate of  claim 6 , wherein the poly(ethylene glycol) is terminally capped with methoxy. 
     
     
         9 . The conjugate of  claim 2  or  3 , wherein the water-soluble polymer has a total weight-average molecular weight in the range of from greater than 5,000 Daltons to about 150,000 Daltons. 
     
     
         10 . The conjugate of  claim 6 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 6,000 Daltons to about 100,000 Daltons. 
     
     
         11 . The conjugate of  claim 10 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 10,000 Daltons to about 85,000 Daltons. 
     
     
         12 . The conjugate of  claim 11 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 20,000 Daltons to about 85,000 Daltons. 
     
     
         13 . The conjugate of  claim 1  or  2 , wherein each water-soluble polymer is linear. 
     
     
         14 . The conjugate of  claim 1  or  2 , wherein each water-soluble polymer is branched. 
     
     
         15 . The conjugate of  claim 1 ,  2  or  3 , wherein the Factor IX moiety is selected from the group consisting of Factor IX, Factor IXa, and biologically active fragments, deletion variants, substitution variants or addition variants of any of the foregoing. 
     
     
         16 . The conjugate of  claim 15 , wherein the Factor IX moiety is Factor IX. 
     
     
         17 . The conjugate of  claim 16 , wherein the Factor IX moiety is Factor IXa. 
     
     
         18 . The conjugate of  claim 6 , wherein the Factor IX moiety is recombinantly derived. 
     
     
         19 . The conjugate of  claim 6 , wherein the Factor IX moiety is blood-derived. 
     
     
         20 . The conjugate of  claim 6 , wherein not more than three water-soluble polymers are attached to the Factor IX moiety. 
     
     
         21 . A composition comprising a plurality of conjugates, wherein at least about 80% of all conjugates in the composition are each comprised of a Factor IX moiety covalently attached to one, two, three or four water-soluble polymers, and further wherein for each water-soluble polymer in the conjugate, the Factor IX moiety is attached to the water-soluble polymer either directly or through a spacer moiety comprised of one or more atoms. 
     
     
         22 . A composition comprising a plurality of conjugates, wherein at least about 85% of all conjugates in the composition each has one to three water-soluble polymers covalently attached to a Factor IX moiety, wherein each water-soluble polymer has a weight-average molecular weight in the range of greater than 5,000 Daltons to about 150,000 Daltons. 
     
     
         23 . A composition comprising a plurality of monoPEGylated Factor IX moiety conjugates, wherein at least about 85% of all conjugates in the composition are monoPEGylated Factor IX moiety conjugates. 
     
     
         24 . The composition of  claim 21 ,  22  or  23 , wherein the water-soluble polymer in each conjugate is selected from the group consisting of a poly(alkylene oxide), poly(vinyl pyrrolidone), poly(vinyl alcohol), polyoxazoline, and poly(acryloylmorpholine). 
     
     
         25 . The composition of  claim 24 , wherein each water-soluble polymer is a poly(alkylene oxide). 
     
     
         26 . The composition of  claim 25 , wherein each poly(alkylene oxide) is a poly(ethylene glycol). 
     
     
         27 . The composition of  claim 26 , wherein the poly(ethylene glycol) is terminally capped with an end-capping moiety selected from the group consisting hydroxy, alkoxy, substituted alkoxy, alkenoxy, substituted alkenoxy, alkynoxy, substituted alkynoxy, aryloxy and substituted aryloxy. 
     
     
         28 . The composition of  claim 26 , wherein the poly(ethylene glycol) is terminally capped with methoxy. 
     
     
         29 . The composition of  claim 21  or  23 , wherein the water-soluble polymer has a total weight-average molecular weight in the range of from greater than 5,000 Daltons to about 150,000 Daltons. 
     
     
         30 . The composition of  claim 26 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 6,000 Daltons to about 100,000 Daltons. 
     
     
         31 . The composition of  claim 30 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 10,000 Daltons to about 85,000 Daltons. 
     
     
         32 . The composition of  claim 31 , wherein the poly(ethylene glycol) has a total weight-average molecular weight in the range of from about 20,000 Daltons to about 85,000 Daltons. 
     
     
         33 . The composition of  claim 6 , wherein each water-soluble polymer is linear. 
     
     
         34 . The composition of  claim 6 , wherein each water-soluble polymer is branched. 
     
     
         35 . The composition of  claim 21 ,  22  or  23 , wherein the Factor IX moiety is selected from the group consisting of Factor IX, Factor IXa, and biologically active fragments, deletion variants, substitution variants or addition variants of any of the foregoing. 
     
     
         36 . The composition of  claim 35 , wherein the Factor IX moiety is Factor IX. 
     
     
         37 . The composition of  claim 35 , wherein the Factor IX moiety is Factor IXa. 
     
     
         38 . The composition of  claim 26 , wherein the Factor IX moiety is recombinantly derived. 
     
     
         39 . The composition of  claim 26 , wherein the Factor IX moiety is blood-derived. 
     
     
         40 . The composition of  claims 21  or  22 , wherein not more than three water-soluble polymers are attached to the Factor IX moiety. 
     
     
         41 . The composition of  claim 26 , wherein the composition is substantially free of albumin. 
     
     
         42 . The composition of  claim 26 , wherein the composition is substantially free of proteins that do not have Factor IX activity. 
     
     
         43 . The composition of  claim 26 , wherein the composition is substantially free of noncovalently attached water-soluble polymers. 
     
     
         44 . The composition of  claim 26 , in lyophilized form. 
     
     
         45 . The composition of  claim 26 , in the form of a liquid. 
     
     
         46 . The composition of  claim 26 , further comprising a pharmaceutically acceptable excipient. 
     
     
         47 . The composition of  claim 23 , wherein each monoPEGylated Factor IX moiety conjugate comprises Factor IX moiety covalently attached, either directly or through a spacer moiety comprised of one or more atoms, to water-soluble polymer. 
     
     
         48 . The composition of  claim 21 ,  22  or  26 , wherein each conjugate comprises an amide linkage linking the Factor IX moiety to either the water-soluble polymer or spacer moiety. 
     
     
         49 . The composition of  claim 21 ,  22  or  26  wherein each conjugate comprises a secondary amine linkage linking the Factor IX moiety to either the water-soluble polymer or spacer moiety. 
     
     
         50 . The composition of  claim 21 ,  22  or  26 , wherein each conjugate comprises a carbamate linkage linking the Factor IX moiety to either the water-soluble polymer or spacer moiety. 
     
     
         51 . The composition of  claim 21 ,  22  or  26 , wherein each conjugate comprises a thioether linkage linking the Factor IX moiety to either the water-soluble polymer or spacer moiety. 
     
     
         52 . The composition of  claim 21 ,  22  or  26 , wherein each conjugate comprises a disulfide linkage linking the Factor IX moiety to either the water-soluble polymer or spacer moiety. 
     
     
         53 . A method for making a conjugate comprising contacting, under conjugation conditions, a Factor IX moiety with a polymeric reagent. 
     
     
         54 . The method of  claim 53 , wherein additional alcohol is not added. 
     
     
         55 . The method of  claim 53 , wherein an oxidizing step is not performed. 
     
     
         56 . A method for making a conjugate comprising adding a polymeric reagent composition to a Factor IX composition under conditions sufficient to result in a conjugate comprising a Factor IX moiety covalently attached, either directly or through a spacer moiety comprised of one or more atoms, to a water-soluble polymer. 
     
     
         57 . The method of  claim 56 , wherein additional alcohol is not added. 
     
     
         58 . The method of  claim 56 , wherein an oxidizing step is not performed. 
     
     
         59 . A method for delivering a conjugate, the method comprising administering to a patient a composition of  claim 46 . 
     
     
         60 . The method of  claim 59 , carried out by subcutaneous injection. 
     
     
         61 . The method of  claim 59 , wherein the patient is suffering from hemophilia B. 
     
     
         62 . The method of  claim 59 , wherein the patient is administered the composition within two days prior to undergoing surgery.