Conjugates of biologically active molecules to functionalized polymers

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a TNF inhibitor or a derivative thereof.

A method of making a TNF inhibitor conjugate, said method comprising reacting a TNF inhibitor or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with a TNF inhibitor or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with a TNF inhibitor or said derivative or a group reactive with a TNF inhibitor or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with a TNF inhibitor or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a TNF inhibitor or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with a TNF inhibitor or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with a TNF inhibitor or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a TNF inhibitor, a derivative of a TNF inhibitor, a biologic other than a TNF inhibitor, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a TNF inhibitor or a derivative of a TNF inhibitor.

A method of making a TNF inhibitor conjugate, said method comprising reacting a TNF inhibitor or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with a TNF inhibitor or said derivative or a group reactive with a TNF inhibitor or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with a TNF inhibitor or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with a TNF inhibitor or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a TNF inhibitor, a derivative of a TNF inhibitor, a biologic other than a TNF inhibitor, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a TNF inhibitor or a derivative of a TNF inhibitor.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a TNF inhibitor or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a TNF inhibitor or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with a TNF inhibitor or a derivative thereof.

A method of making an insulin conjugate, said method comprising reacting insulin or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with insulin or said derivative thereof:

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with insulin or derivative thereof or is a group reactive with insulin or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with a insulin or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with insulin or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is insulin or a derivative thereof.

A method of making an insulin conjugate, said method comprising reacting insulin or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with insulin or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with insulin or said derivative or a group reactive with insulin or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with insulin or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with insulin or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with insulin or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with insulin or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently insulin, a derivative of insulin, a biologic other than insulin, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is insulin or a derivative of insulin.

A method of making an insulin conjugate, said method comprising reacting insulin or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with insulin or said derivative or a group reactive with insulin or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with insulin or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with insulin or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently insulin, a derivative of insulin, a biologic other than insulin, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is insulin or a derivative of insulin.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is insulin or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with insulin or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with insulin or a derivative thereof.

A method of making an Omalizumab conjugate, said method comprising reacting Omalizumab or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with Omalizumab or said derivative thereof:

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with Omalizumab or derivative thereof or is a group reactive with Omalizumab or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with Omalizumab or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with Omalizumab or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is Omalizumab or a derivative thereof.

A method of making an Omalizumab conjugate, said method comprising reacting Omalizumab or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with Omalizumab or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with Omalizumab or said derivative or a group reactive with Omalizumab or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with Omalizumab or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with Omalizumab or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with Omalizumab or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with Omalizumab or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently Omalizumab, a derivative of Omalizumab, a biologic other than Omalizumab, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is Omalizumab or a derivative of Omalizumab.

A method of making an Omalizumab conjugate, said method comprising reacting Omalizumab or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with Omalizumab or said derivative or a group reactive with Omalizumab or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with Omalizumab or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with Omalizumab or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently Omalizumab, a derivative of Omalizumab, a biologic other than Omalizumab, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is Omalizumab or a derivative of Omalizumab.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is Omalizumab or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with Omalizumab or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with Omalizumab or a derivative thereof.

A method of making an a clotting factor conjugate, said method comprising reacting a clotting factor or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with a clotting factor or said derivative thereof:

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a clotting factor or derivative thereof or is a group reactive with a clotting factor or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with a a clotting factor or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a clotting factor or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a clotting factor or a derivative thereof.

A method of making a clotting factor conjugate, said method comprising reacting a clotting factor or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with a clotting factor or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with a clotting factor or said derivative or a group reactive with a clotting factor or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with a clotting factor or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a clotting factor or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with a clotting factor or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with a clotting factor or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a clotting factor, a derivative of a clotting factor, a biologic other than a clotting factor, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a clotting factor or a derivative of a clotting factor.

A method of making a clotting factor conjugate, said method comprising reacting a clotting factor or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with a clotting factor or said derivative or a group reactive with a clotting factor or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with a clotting factor or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with a clotting factor or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a clotting factor, a derivative of a clotting factor, a biologic other than a clotting factor, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a clotting factor or a derivative of a clotting factor.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a clotting factor or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a clotting factor or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with a clotting factor or a derivative thereof.

A method of making a polypeptide that boosts red or white cell production conjugate, said method comprising reacting a polypeptide that boosts red or white cell production or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with a polypeptide that boosts red or white cell production or said derivative thereof:

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or derivative thereof or is a group reactive with a polypeptide that boosts red or white cell production or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with a a polypeptide that boosts red or white cell production or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a polypeptide that boosts red or white cell production or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a polypeptide that boosts red or white cell production or a derivative thereof.

A method of making a polypeptide that boosts red or white cell production conjugate, said method comprising reacting a polypeptide that boosts red or white cell production or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with a polypeptide that boosts red or white cell production or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or said derivative or a group reactive with a polypeptide that boosts red or white cell production or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a polypeptide that boosts red or white cell production or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with a polypeptide that boosts red or white cell production or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a polypeptide that boosts red or white cell production, a derivative of a polypeptide that boosts red or white cell production, a biologic other than a polypeptide that boosts red or white cell production, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a polypeptide that boosts red or white cell production or a derivative of a polypeptide that boosts red or white cell production.

A method of making a polypeptide that boosts red or white cell production conjugate, said method comprising reacting a polypeptide that boosts red or white cell production or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or said derivative or a group reactive with a polypeptide that boosts red or white cell production or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with a polypeptide that boosts red or white cell production or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a polypeptide that boosts red or white cell production, a derivative of a polypeptide that boosts red or white cell production, a biologic other than a polypeptide that boosts red or white cell production, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a polypeptide that boosts red or white cell production or a derivative of a polypeptide that boosts red or white cell production.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a polypeptide that boosts red or white cell production or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a polypeptide that boosts red or white cell production or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with a polypeptide that boosts red or white cell production or a derivative thereof.

A method of making an antibody conjugate, said method comprising reacting an antibody or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with an antibody or said derivative thereof:

• or a salt thereof, wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with an antibody or derivative thereof or is a group reactive with an antibody or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with an antibody or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with an antibody or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is an antibody or a derivative thereof.

A method of making an antibody conjugate, said method comprising reacting an antibody or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with an antibody or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with an antibody or said derivative or a group reactive with an antibody or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with an antibody or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with an antibody or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with an antibody or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with an antibody or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently an antibody, a derivative of an antibody, a biologic other than an antibody, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is an antibody or a derivative of an antibody.

A method of making an antibody conjugate, said method comprising reacting an antibody or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with an antibody or said derivative or a group reactive with an antibody or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with an antibody or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with an antibody or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently an antibody, a derivative of an antibody, a biologic other than an antibody, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is an antibody or a derivative of an antibody.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is an antibody or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with an antibody or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with an antibody or a derivative thereof.

A method of making a biologically active molecule conjugate, said method comprising reacting a biologically active molecule or a derivative thereof with a preparation comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (1) under conditions suitable for group M or R to react with a biologically active molecule or said derivative thereof:

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a biologically active molecule or derivative thereof or is a group reactive with a biologically active molecule or derivative thereof;
• R is absent or selected from the group consisting of:
• hydrogen, a protecting group, a hydrophobic separation handle, or an activating group;
• R1 is absent or a hydrophobic separation handle;
• wherein when M is a protected group that when deprotected is reactive with a biologically active molecule or derivative thereof, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a biologically active molecule or derivative thereof, R is absent, hydrogen, or an activating group;
• and
• wherein only one of R, R1, and G can be a hydrophobic separation handle.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• R1 is absent or a hydrophobic separation handle, wherein only one of R1 and G can be a hydrophobic separation handle;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a biologically active molecule or a derivative thereof.

A method of making a biologically active molecule conjugate, said method comprising reacting a biologically active molecule or a derivative thereof with a preparation comprising a compound of formula (2) under conditions suitable for group M or R to react with a biologically active molecule or said derivative thereof:

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M and M1 are independently selected from a protected group that when deprotected is reactive with a biologically active molecule or said derivative or a group reactive with a biologically active molecule or said derivative, wherein M and M1 are different;
• and
• R and R1 are independently absent, hydrogen, a protecting group, or an activating group;
• wherein when M is a protected group that when deprotected is reactive with a biologically active molecule or said derivative, then R is a protecting group or a hydrophobic separation handle;
• wherein when M is a group reactive with a biologically active molecule or said derivative, R is absent, hydrogen, or an activating group;
• wherein when M1 is a protected group that when deprotected is reactive with a biologically active molecule or said derivative, then R1 is a protecting group or a hydrophobic separation handle;
• and
• wherein when M1 is a group reactive with a biologically active molecule or said derivative, R1 is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a structure of formula (10):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein each linking group is bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K and K1 are independently selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G and G1 are independently absent or are selected from the group consisting of:
• alkoxy and a hydrophobic separation handle;
• each pair of Z1 and Z2 and Z3 and Z4 are independently selected from O and NH, wherein only one of each pair of Z1 and Z2 and Z3 and Z4 can be NH;
• L and L1 are independently selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L3 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a biologically active molecule, a derivative of a biologically active molecule, a biologic other than a biologically active molecule, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a biologically active molecule or a derivative of a biologically active molecule.

A method of making a biologically active molecule conjugate, said method comprising reacting a biologically active molecule or a derivative thereof with a preparation comprising a compound of formula (3):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is selected from a protected group that when deprotected is reactive with a biologically active molecule or said derivative or a group reactive with a biologically active molecule or said derivative;
• M2 is selected from O, S or NH;
• and
• R is absent, a protecting group, a hydrophobic separation handle, or an activating group;
• R2 is hydrogen or a protecting group;
• wherein when M is a protected group that when deprotected is reactive with a biologically active molecule or said derivative, then R is a protecting group or a hydrophobic separation handle;
• and
• wherein when M is a group reactive with a biologically active molecule or said derivative, R is absent, hydrogen, or an activating group.

A conjugate, or a pharmaceutically acceptable salt thereof, comprising a compound of formula (11):

• or a salt form thereof,
• wherein:
• polymer is a linear, water-soluble, non-peptidic, and non-nucleotidic polymer backbone, wherein M2 and the phosphonate-derived functional group are bonded at a different terminus of said polymer;
• E and E1 are independently O or S;
• K is selected from the group consisting of:
• alkylene, alkyleneoxyalkylene, and oligomeric alkyleneoxyalkylene;
• G is selected from the group consisting of:
• hydrogen, alkoxy, and a hydrophobic separation handle;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• L4 is a covalent linking moiety between L on the polymer backbone and B1;
• and
• B and B1 are independently a biologically active molecule, a derivative of a biologically active molecule, a biologic other than a biologically active molecule, a drug, a detectable group, a separation moiety, wherein at least one of B and B1 is a biologically active molecule or a derivative of a biologically active molecule.

A method of preparing a compound comprising a water-soluble, non-peptidic, and non-nucleotidic polymer backbone having at least one terminus covalently bonded to a structure of formula (9):

• or a salt thereof,
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• L2 is a covalent linking moiety between L on the polymer backbone and B;
• and
• B is a biologically active molecule or a derivative thereof;
• comprising the steps of:
• (a) providing a composition comprising a compound of formula (8):
• wherein:
• A is the point of covalent bonding to the terminus of the polymer backbone;
• E is O or S;
• Y represents an optionally substituted residue selected from alkyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl;
• Z1 and Z2 are independently selected from O and NH, wherein only one of Z1 and Z2 can be NH;
• L is selected from the group consisting of:
• a divalent radical of a nucleoside, alkylene, alkyleneoxyalkylene, oligomeric alkyleneoxyalkylene, and unsubstituted and substituted arylene;
• M is a protected group that when deprotected is reactive with a biologically active molecule or a derivative thereof;
• R is a hydrophobic separation handle;
• R1 is absent or a hydrophobic separation handle;
• and
• (b) removing the hydrophobic separation handle(s);
• (c) optionally reacting the compound obtained in step (b) with an activating agent;
• and
• (d) reacting the compound obtained in step (b), or, optionally in step (c), with a biologically active molecule or a derivative thereof.