PURPOSE: We purposed to design a cationic polymer that binds to pDNA to form polyplexes and that subsequently degrades within a few days at physiological pH and temperature, releasing the DNA in the cytosol of a cell. METHODS: We synthesized a new monomer carbonic acid 2-dimethylamino-ethyl ester 1-methyl-2-(2-methacryloylamino)-ethyl ester (abbreviated HPMA-DMAE) and the corresponding polymer. Hydrolysis of the carbonate ester of both the monomer and the polymer was investigated at 37 degrees C. The DNA condensing properties of the pHPMA-DMAE was studied using dynamic light scattering (DLS) and zeta potential measurements. Degradation of the polyplexes at 37 degrees C and pH 7.4 was monitored with DLS and gel electrophoresis. In vitro transfections were performed in COS-7 cell line. RESULTS: pHPMA-DMAE is able to condense DNA into small particles (110 nm) with a positive zeta potential. The half-life of the polymer and monomer at 37 degrees C and pH 7.4 was around 10 h whereas at pH 5, the half-life was 380 h. In line with this, due to hydrolysis of the side groups, pHPMA-DMAE-based polyplexes dramatically increased in size at 37 degrees C and pH 7.4 whereas at pH 5.0, only a very small increase was observed. Interestingly, intact DNA was released from the polyplexes after 48 h at pH 7.4 whereas all DNA remained bound to the polymer at pH 5.0. Polyplexes were able to transfect cells with minimal cytotoxicity if the endosomal membrane-disrupting peptide INF-7 was added to the polyplex formulation. CONCLUSIONS: Degradation of the cationic side-chains of a polymer is a new tool for time-controlled release of DNA from polyplexes, preferably within the cytosol and/or nucleus.
PURPOSE: We purposed to design a cationic polymer that binds to pDNA to form polyplexes and that subsequently degrades within a few days at physiological pH and temperature, releasing the DNA in the cytosol of a cell. METHODS: We synthesized a new monomer carbonic acid 2-dimethylamino-ethyl ester 1-methyl-2-(2-methacryloylamino)-ethyl ester (abbreviated HPMA-DMAE) and the corresponding polymer. Hydrolysis of the carbonate ester of both the monomer and the polymer was investigated at 37 degrees C. The DNA condensing properties of the pHPMA-DMAE was studied using dynamic light scattering (DLS) and zeta potential measurements. Degradation of the polyplexes at 37 degrees C and pH 7.4 was monitored with DLS and gel electrophoresis. In vitro transfections were performed in COS-7 cell line. RESULTS:pHPMA-DMAE is able to condense DNA into small particles (110 nm) with a positive zeta potential. The half-life of the polymer and monomer at 37 degrees C and pH 7.4 was around 10 h whereas at pH 5, the half-life was 380 h. In line with this, due to hydrolysis of the side groups, pHPMA-DMAE-based polyplexes dramatically increased in size at 37 degrees C and pH 7.4 whereas at pH 5.0, only a very small increase was observed. Interestingly, intact DNA was released from the polyplexes after 48 h at pH 7.4 whereas all DNA remained bound to the polymer at pH 5.0. Polyplexes were able to transfect cells with minimal cytotoxicity if the endosomal membrane-disrupting peptide INF-7 was added to the polyplex formulation. CONCLUSIONS: Degradation of the cationic side-chains of a polymer is a new tool for time-controlled release of DNA from polyplexes, preferably within the cytosol and/or nucleus.
Authors: Themis R Kyriakides; Charles Y Cheung; Niren Murthy; Paul Bornstein; Patrick S Stayton; Allan S Hoffman Journal: J Control Release Date: 2002-01-17 Impact factor: 9.776
Authors: J Luten; J H van Steenis; R van Someren; J Kemmink; N M E Schuurmans-Nieuwenbroek; G A Koning; D J A Crommelin; C F van Nostrum; W E Hennink Journal: J Control Release Date: 2003-05-20 Impact factor: 9.776
Authors: Thorsten Bieber; Wolfgang Meissner; Sawa Kostin; Axel Niemann; Hans-Peter Elsasser Journal: J Control Release Date: 2002-08-21 Impact factor: 9.776
Authors: Arjen M Funhoff; Cornelus F van Nostrum; Gerben A Koning; Nancy M E Schuurmans-Nieuwenbroek; Daan J A Crommelin; Wim E Hennink Journal: Biomacromolecules Date: 2004 Jan-Feb Impact factor: 6.988
Authors: Thomas A Werfel; Corban Swain; Christopher E Nelson; Kameron V Kilchrist; Brian C Evans; Martina Miteva; Craig L Duvall Journal: J Biomed Mater Res A Date: 2016-01-11 Impact factor: 4.396
Authors: Bo Lou; Kate Connor; Kieron Sweeney; Ian S Miller; Alice O'Farrell; Eduardo Ruiz-Hernandez; David M Murray; Garry P Duffy; Alan Wolfe; Enrico Mastrobattista; Annette T Byrne; Wim E Hennink Journal: Drug Deliv Transl Res Date: 2019-06 Impact factor: 4.617
Authors: Xulin Jiang; Aschwin van der Horst; Mies J van Steenbergen; Niels Akeroyd; Cornelus F van Nostrum; Peter J Schoenmakers; Wim E Hennink Journal: Pharm Res Date: 2006-02-21 Impact factor: 4.200