Three-branched polyethylene glycol-G-CSF conjugate
Inventors
Jo, Yeong-Woo • Yoo, Won-Young • Jeon, Hyun-Kyu • Choi, Yun-Kyu • Jang, Hye-In • Kim, Byong-Moon • Lee, Sung-hee • Kang, Soo-Hyung • Yoo, Moo-Hi
Assignees
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Publication Number
US-8329191-B2
Publication Date
2012-12-11
Expiration Date
Abstract
The present invention relates to three-branched PEG-G-CSF conjugate of general formula (1) in which the bonding ratio of three-branched polyethylene glycol (PEG) and G-CSF is 1:1 (mol/mol), wherein PEG has an average molecular weight of from 200 to 45,000 daltons; a pharmaceutical composition comprising the same, and a preparing method thereof.
Core Innovation
The disclosure describes a three-branched PEG-G-CSF conjugate having a defined general formula (1), in which the bonding ratio of three-branched polyethylene glycol (PEG) and G-CSF is 1:1 (mol/mol) and PEG has an average molecular weight of about 23,000 daltons. The conjugate includes linker group Z defined as (CH2)S or (CH2)S NHCO(CH2)S, where S is an integer of 1 to 6, and Y is an amide bond formed by combining the NH2 functional group in G-CSF and a functional group of a PEG derivative.
The disclosure further defines that n is an integer of 1 to 1,000 and m is an integer of 10 to 1,000 for the conjugate. It uses a PEG derivative structure featuring a three-branched skeleton with three OH regions on a glycerol skeleton, and end-group activation while other regions are monomethoxylated. The PEG derivative functional group includes N-hydroxysuccinimide as a protein/peptide-reactive functional group, enabling formation of the Y amide bond with the NH2 functional group in G-CSF.
The invention addresses neutropenia and related conditions by providing a three-branched PEG-G-CSF conjugate and pharmaceutical compositions comprising the conjugate together with pharmaceutically acceptable components. Experimental support in the disclosure describes mono three-branched PEG-G-CSF conjugates with PEG molecular weights of 23,000 and 43,000 daltons, confirmation of position-isomer ratios, markedly increased pharmacokinetic exposure compared with native G-CSF, and stability behavior indicating reduced PEG shedding and reduced aggregation compared with linear or two-branched comparators.
In vivo results described in the disclosure include increased neutrophil proliferation and rapid recovery of neutrophil counts in a cyclophosphamide-induced neutropenia mouse model.
Claims Coverage
The document contains two independent claims. Independent claim coverage centers on a three-branched PEG-G-CSF conjugate with a defined 1:1 PEG:G-CSF bonding ratio and an amide linkage (Y) formed between G-CSF NH2 and a PEG derivative functional group (Y), with specified linker group Z and integer ranges for n and m.
Three-branched PEG-G-CSF conjugate with 1:1 bonding ratio and defined linker and amide bond
A three-branched PEG-G-CSF conjugate of general formula (1) in which the bonding ratio of three-branched PEG and G-CSF is 1:1 (mol/mol) and PEG has an average molecular weight of about 23,000 daltons, wherein n is an integer of 1 to 1,000; m is an integer of 10 to 1,000; Z is (CH2)S or (CH2)S NHCO(CH2)S as a linker with S being an integer of 1 to 6; and Y is an amide bond formed by combining the NH2 functional group in G-CSF and a functional group of the PEG derivative.
Pharmaceutical composition comprising the defined three-branched PEG-G-CSF conjugate
A pharmaceutical composition comprising a three-branched PEG-G-CSF conjugate of general formula (1) in which the bonding ratio of three-branched PEG and G-CSF is 1:1 (mol:mol) and PEG has an average molecular weight of 23,000 daltons, wherein n is an integer of 1 to 1,000; m is an integer of 10 to 1,000; Z is (CH2)S or (CH2)S as a linker with S being an integer of 1 to 6; Y is an amide bond formed by combining the NH2 functional group in G-CSF and a functional group of the PEG derivative, and at least one pharmaceutically acceptable diluent, antiseptic, solubilizer, emulsifier, adjuvant, or combination thereof.
Across the independent claims, the central inventive subject matter is the defined three-branched PEG-G-CSF conjugate (specified PEG:G-CSF bonding ratio, PEG molecular weight, linker Z, and amide bond Y) and, in the second independent claim, the inclusion of at least one pharmaceutically acceptable diluent or related formulation component.
Stated Advantages
Documented Applications
Treatment of neutropenia using a pharmaceutical composition comprising the defined three-branched PEG-G-CSF conjugate, where the dependent claim specifies neutropenia caused by cyclophosphamide treatment.
In vivo use in a cyclophosphamide-induced neutropenia mouse model is described, including neutrophil proliferation activity and rapid recovery of neutrophils.
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