DOCSLIB.ORG

Base Strength-Reactivity Effects in Polyethylenimine Esterolysis Reactions

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Base Strength-Reactivity Effects in Polyethylenimine Esterolysis Reactions BASE STRENGTH-REACTIVITY EFFECTS IN POLYETHYLENIMINE ESTEROLYSIS REACTIONS BY CURTIS S. LEGE A DISSERTATION PRESENTED TO THE GRADUATE COUNCIL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY UNIVERSITY OF FLORIDA 1979 TO MY WIFE AND DAUGHTER JOE ANN AND SPRING ACKNOWLEDGMENTS The author wishes to express his appreciation to his research director, James A. Deyrup, for his thoughtful guidance and criticism as well as his encouragement of individual creativity in problem selection and pursuit. The author is further indebted to Professor John A. Zoltewicz for the many helpful discussions and use of equipment. The author wishes to thank the faculty and staff of the University of Florida and his fellow graduate students for an enjoyable and fruitful experience. Special thanks are extended to Tom Baugh and Eric Langenmeyer for their friendship and advice. The author is especially indebted to his wife who has provided not only encouragement throughout but also has made considerable contribution to the preparation of this dissertation. TABLE OF CONTENTS Page ACKNOWLEDGMENTS _ _ m LIST OF TABLES vii LIST OF FIGURES viii ABSTRACT x CHAPTER I POLYMER ESTEROLYSIS REACTIONS: DISCUSSION AND PROPOSAL 1 Introduction \ Discussion 2 Mechanism of Polymer Esterolysis Reactions 2 Polymer Catalyzed Hydrolysis 8 Summary n Proposal 12 Introduction 12 Nucleophiles 12 Polymer 13 Ester 15 Results 16 CHAPTER II AMINOLYSIS OF P-NITROPHENYL ESTERS BY DODECYL POLYETHYLENIMINE 2 2 Results and Discussion 22 Characterization 22 Basicity of PEI-D-NH -HC1 2 25 Kinetic Analysis....; 26 Eronsted Relationship 34 Summary 43 IV CHAPTER III HYDROLYSIS OF P-NITROPHENYL ESTERS CATALYZED BY POLYMER BOUND HETEROCYCLES . 44 Introduction 44 Polyethylenimine Systems 44 Preparation 44 Primary Amine Analysis 45 Further Kinetic Analysis 49 Summary 51 Non Polyethylenimine Homopolymer Systems. 52 Conclusion 58 CHAPTER IV EXPERIMENTAL 60 Introduction 60 Syntheses 61 Dodecyl Polyethylenimine PEI-D 61 Dodecyl Polyethylenimine Hydro- chloride PEI-D-NH2-HC1 61 Dodecyl- 4 -me thy lenepyridine-poly- ethylenimine Hydrochloride PEI-D-Pyr-HCl 62 Dodecyl-N- (2-pyridyl) -3-propylamine- polyethylenimine Hydrochloride PEI-D-APyr-HCl. 62 Dodecyl-4 (5) -methylenimidazole-poly- ethylenimine Hydrochloride PEI-D-Im-HCl 63 Dodecyl-4 (5) -methylenimidazole- isopropyl-polyethylenimine Hydrochloride PEI-D-Im-Ip-HCl . 64 Dodecyl-4 (5) -methylenimidazole-iso- propyl-isopropyl-polyethylinimine ^Hydrochloride PEI-D-Im-Ip2-HCl. 65 4(d) -Chloromethylimidazole Hydro- chloride 65 4(5) -Hydroxymethylimidazole Hydrochloride 65 3-Bromopropanal Dimethyl Acetal 66 N- (2-pyridyl) -3-aminopropanal Dimethyl Acetal 6 6 N- (2-pyridyl) -3-aminopropanal Hydrochloride 66 p-Nitrophenyl Acetate PNPA 67 p-Nitrophenol Caproate PNPC 67 Composition of PEI Derivatives by Elemental Analysis 6 7 Composition of PEI Derivatives by NMR Spectral Analysis 7 3 Determination of Primary Content of PEI Derivatives Using Trinitro- benzene Sulfonate (TNBS) 73 Primary Amine Detection by Ninhydrin 7 4 Relative Concentrations of Imidazole Containing Polymer Solutions 75 Potentiometric Titration of PEI-D- NH -HC1 2 76 Preparation and Standardization of C0 Free 0.1 N KOH Titrant 2 7 8 Preparation of Ester Solutions Used for Kinetic Studies 78 Preparation of Polymer Solutions for Kinetic Studies 78 Buffer Solutions 79 Kinetic Method 79 Background Rate 8 Pyridine Ionization in PEI-D-Pyr-HCl 81 APPENDIX. 33 REFERENCES % 9 BIOGRAPHICAL SKETCH 94 VI . LIST OF TABLES Table Page II-l PRIMARY AMINE AND ISOPROPYL CONTENT OF PEI-D-NH -HC1 AND 2 PEI-D-IpHCl 2 3 III-l % PEI UNITS BOUND TO HETEROCYCLES 45 IV-1 COMPOSITION OF PEI DERIVATIVES FROM ELEMENTAL ANALYSIS DATA , 70 IV- 2 FRACTION OF PEI UNITS ALKYLATED FROM NMR SPECTRAL DATA 72 IV- 3 PRIMARY AMINE CONTENT OF PEI DERIVATIVES USING TNBS 74 IV-4 POTENTIOMETRIC TITRATION DATA FOR PEI-D-NH - HC1 2 77 IV- 5 ESTEROLYSIS RATES FOR PEI-D-NH -HC1 83 IV-6 ESTEROLYSIS RATES FOR PEI-D-Ip-HCl 84 IV- 7 ESTEROLYSIS RATES FOR PEI-D-Im-HCl 84 IV- 8 ESTEROLYSIS RATES FOR PEI-D-Im-Ip-HCl 85 2 IV- 9 ESTEROLYSIS RATES FOR PEI-D-Im-Ip -HCl 85 IV-10 ESTEROLYSIS RATES FOR PEI-D-APyr-HCl 86 IV- 11 ESTEROLYSIS RATES FOR PEI-D-Pyr-HCl 87 IV- 12 ESTEROLYSIS RATES • FOR ETHYLENEDIAMINE HC1 . 87 Vll LIST OF FIGURES Figure p age I-la Solvolysis of PNPA catalyzed by Poly- 4 (5) -vinylimadazole and imidazole 10 I-lb Bifunctional mechanisms proposed for poly-4 (5) -vinylimidazole catalyzed hydrolysis of PNPA 10 1-2 Esterolysis of PNPA by PEI-D-Pyr-KCl 18 1-3 Esterolysis of PNPA by PEI-D-APyr-HCl . 19 1-4 Esterolysis of PNPA by PEI-D-Im-HCl 20 1-5 Esterolysis of PNPA by PEI-D-NH--HC1 compared to esterolysis of PNPA by PEI bound heterocyclic systems 21 II-l Comparison of pK a aDp dependence on a for PEI-D-NH -HC1 with low molecular weight polyamines 27 II-2 Comparison of esterolysis rates for PEI-D-NH -KC1 and PEI-D-Ip-HCl 29 2 II-3 Inhibition of ethylenediamine, PEI-D-NH2- HC1 (pH=7.00) and PEI-D-NH^-HCl (pH=8.82) esterolysis of PNPA by p-nitrophenol anion (PNP) 31 II-4 Plot of log(k /a) against pH for PEI-D- NH 2 -HC1 esterolysis of PNPC and PNPA 33 II-5 Bronsted plot for the esterolysis of PNPA by simple primary and cyclic secondary amines 36 II-6 Bronsted plot for the esterolysis of PNPA by imidazoles and anilines 37 II-7 Plot of pK vs pH for PEI-D-NH„-HC1 38 a , app 2. II-8 Bronsted plot for the esterolysis of PNPA and PNPC by PEI-D-NH -HCl 4 II-9 plot of e vs pK or [a] for the esterolysis of PNPC and PNPA by PEI-D-NH -HCl 41 III-l Effects of repetitive isopropylation of PEI-D- Im-HCl on PNPA esterolysis reactivity 48 III-2 Comparison of PNPA esterolysis rates for PEI-D-HC1 and PEI-D-Ip-HCl 50 III-3 Plot of kobs against a for the esterolysis of 2, 4-dinitrophenyl acetate by poly-4-vinylpyri- dine and pyridine 54 III-4 Plot of k„ against a for the esterolysis of PNPA by PEI-D-NH -HC1 55 2 III-5 Bronsted plot for the esterolysis of PNPA by poly-4 (5) -vinylimidazole and imidazoles 56 III-6 Bronsted plot for the esterolysis of 2,4-dini- trophenylacetate by poly- (4) -vinylpyridine. 5 7 IV-1 Typical spectrum of a PEI-D derivative (PEI-D- APyr-HCl) 71 Abstract of Dissertation Presented to the Graduate Council of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy BASE STRENGTH-REACTIVITY EFFECTS IN POLYETHYLENIMINE ESTEROLYSIS REACTIONS BY CURTIS S. LEGE December 1979 Chairman: James A. Deyrup Major Department: Chemistry The esterolysis of p-nitrophenyl acetate (PNP) and p-nitrophenyl caproate (PNPC) by polyethylenimine (PEI) derivatives was studied as a function of pH . The reactivity of partly dodecylated PEI containing primary amines (PEI-D-NH -HC1) , imidazole (PEI-D-Im-HCl) 2 , pyridine (PEI-D-Pyr-HCl) , and 2-aminopyridine (PEI-D-APyr-HCl) was found to be pH dependent without exception. The PEI-D-NH ~HC1 system was found to react 2 with PNPA predomi- nantly by nucleophilic attack of polymeric primary amine. Potentiometric titration showed the pK or PEI-D-NH„-HC1 a , app 2 also to be dependent. A Bronsted-like plot of log k for polymeric free primary amine aaainst pK for the ester- a , app olysis of PNPA with PEI-D-NH -HC1 had a slope (3 value) of 0.81. A Bronsted plot of a series of low molecular weight amines with PNPA taken from the literature had a slope of 0.83. The Bronsted-like plot of log k with pK , an a , a p p approach unknown prviously in polymer systems, quantita- tively accounted for the pH dependence of the reaction of PEI-D-NH -HC1 with PNPA. 2 The reactivity of PEI-D-NH -HC1 toward PNPA is dependent on the pK of the nucleophile as in the case of low molecular weight amine nucleophiles . The slope of the plot of log k versus pK for the a ^ esterolysis of PNPC by PEI-D-NH -HC1 had a slope 2 of 1.06. This slope was dissected into a pH dependent polymeric enhancement component (E) and a pH dependent nucleophilic component. The pK a , app of the heterocycles in the heterocycle containing polymers was unobtainable in the pH region of interest. However, the pHdependencies were assigned to nucleophilic effects by analogy. This approach proved useful in reviewing literature reports of other polymeric esterolytic systems. Data for PNPA esterolysis by poly-4 ( 5) vinylimidazole and for 2,4-dinitrophenyl acetate esterolysis were plotted as described above with slopes of 0.8 and 1.3, respectively. CHAPTER I POLYMER ESTEROLYSIS REACTIONS: DISCUSSION AND PROPOSAL Introduction Esterolysis by polymer systems has received considerable attention since the early days of polymer research. This area of research was spurred as the mechanism of enzyme reactions became better understood. It was hoped that synthetic polymer systems might offer a method of modeling or duplicating the action of the naturally occurring polymers (enzymes) . The aspects of enzyme action of greatest interest in this field are high selectivity and high efficiency. To this end synthetic polymer systems with particular binding properties and high reactivity have been sought. One of the aspects of polymer esterolysis reactions which has received a great deal of attention is coopera- tivity, i.e., two or more functional groups in the same polymer molecule participating in ester cleavage.
Load more

Recommended publications
  • Modification of Branched Polyethyleneimine Using Mesquite
    polymers Article Modification of Branched Polyethyleneimine Using Mesquite Gum for Its Improved Hemocompatibility Ana M. Pinilla-Torres 1 , Paola Y. Carrión-García 2, Celia N. Sánchez-Domínguez 2 , Hugo Gallardo-Blanco 3,* and Margarita Sánchez-Domínguez 1,* 1 Grupo de Química Coloidal e Interfacial Aplicada a Nanomateriales y Formulaciones, Centro de Investigación en Materiales Avanzados, S.C. (CIMAV, S.C.), Unidad Monterrey, Apodaca 66628, Mexico; [email protected] 2 Departamento de Bioquímica y Medicina Molecular, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey 64460, Mexico; [email protected] (P.Y.C.-G.); [email protected] (C.N.S.-D.) 3 Departamento de Genética, Facultad de Medicina, Universidad Autónoma de Nuevo León, Monterrey 64460, Mexico * Correspondence: [email protected] (H.G.-B.); [email protected] (M.S.-D.) Abstract: In the present study, the modification of branched polyethyleneimine (b-PEI) was car- ried out using mesquite gum (MG) to improve its hemocompatibility to be used in biomedi- cal applications. In the copolymer synthesis process (carboxymethylated mesquite gum grafted polyethyleneimine copolymer (CBX-MG-PEI), an MG carboxymethylation reaction was initially carried out (carboxymethylated mesquite gum (CBX-MG). Subsequently, the functionalization be- Citation: Pinilla-Torres, A.M.; tween CBX-MG and b-PEI was carried out using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide Carrión-García, P.Y.; (EDC) and N-hydroxysuccinimide (NHS) as crosslinking agents. The synthesis products were char- Sánchez-Domínguez, C.N.; acterized using Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy Gallardo-Blanco, H.; (XPS), and thermogravimetric analysis (TGA). Thermogravimetric analysis showed that CBX-MG Sánchez-Domínguez, M.
    [Show full text]
  • Polyethylenimine-Derived Gene Carriers and Their Complexes with Plasmid DNA
    Polyethylenimine-derived Gene Carriers and their Complexes with plasmid DNA Design, Synthesis and Characterization Dissertation to obtain the Degree of Doctor of Natural Sciences (Dr. rer. nat.) from the Faculty of Chemistry and Pharmacy University of Regensburg Presented by Uta Lungwitz From Greifswald August 2006 To my family This work was carried out from October 2001 until August 2006 at the Department of Pharmaceutical Technology of the University of Regensburg. The thesis was prepared under the supervision of Prof. Dr. Achim Göpferich Submission of the Ph.D. application: July, 12th 2006 Date of examination: August, 10th 2006 Examination board: Chairman: Prof. Dr. S. Elz 1. Expert: Prof. Dr. A. Göpferich 2. Expert: Prof. Dr. A. Buschauer 3. Examiner: Prof. Dr. J. Heilmann Table of Contents Chapter 1 Introduction: Polyethylenimine-based non-viral gene 7 delivery systems Chapter 2 Goal of the thesis 55 Chapter 3 Synthesis of per-N-methylated polyethylenimine and its 61 application for non-viral gene delivery Chapter 4 Synthesis and characterization of poly (2-ethyl-2- 85 oxazoline) and linear polyethylenimine Chapter 5 Low molecular weight polyethylenimine-plasmid DNA 113 polyplexes: Particle properties and transfection efficacy Chapter 6 Degradable low molecular weight linear 147 polyethylenimines for non-viral gene transfer Chapter 7 Methoxy poly (ethylene glycol) – low molecular weight 181 linear polyethylenimine – derived copolymers enable polyplex shielding Chapter 8 Summary and conclusions 211 Appendix Abbreviations 219 Curriculum Vitae List of publications Acknoledgments 1. POLYETHYLENEIMINE-BASED NON- VIRAL GENE DELIVERY SYSTEMS U. Lungwitz, M. Breunig, T. Blunk, A. Göpferich Department of Pharmaceutical Technology, University of Regensburg, D- 93053 Regensburg, Germany European Journal of Pharmaceutics and Biopharmaceutics, 60 (2) 2005 p.
    [Show full text]
  • Analysis of Polyethylenimine by Electrospray Ionization Mass
    ANALYSIS OF POLYETHYLENIMINE BY ELECTROSPRAY IONIZATION MASS SPECTROMETRY AND SIZE EXCLUSION CHROMATOGRAPHY By Jianghong Gu Submitted to the Faculty of the College of Arts and Sciences of American University in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy In Chemi /Dr. Monika Konakli'onaklieva Dean 0 1 the College of Arts and Sciences Dr. Milena Shahu Date 2005 American University Washington, D.C. 20016 AMERICAN UNIVERSITY LIBRARY Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. UMI Number: 3187219 Copyright 2005 by Gu, Jianghong All rights reserved. INFORMATION TO USERS The quality of this reproduction is dependent upon the quality of the copy submitted. Broken or indistinct print, colored or poor quality illustrations and photographs, print bleed-through, substandard margins, and improper alignment can adversely affect reproduction. In the unlikely event that the author did not send a complete manuscript and there are missing pages, these will be noted. Also, if unauthorized copyright material had to be removed, a note will indicate the deletion. ® UMI UMI Microform 3187219 Copyright 2006 by ProQuest Information and Learning Company. All rights reserved. This microform edition is protected against unauthorized copying under Title 17, United States Code. ProQuest Information and Learning Company 300 North Zeeb Road P.O. Box 1346 Ann Arbor, Ml 48106-1346 Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. © COPYRIGHT by Jianghong Gu 2005 ALL RIGHTS RESERVED Reproduced with permission of the copyright owner. Further reproduction prohibited without permission. ANALYSIS OF POLYETHYLENIMINE BY ELECTROSPRAY IONIZATION MASS SPECTROMETRY AND SIZE EXCLUSION CHROMATOGRAPHY By Jianghong Gu ABSTRACT The efficient delivery of therapeutic genes into target cells or tissues is a critical goal in gene therapy.
    [Show full text]
  • Selective Polymerization of 2-Isopropenyl-2-Oxazoline and Cross-Linking Reaction of the Polymers
    Polymer Journal, Vol. 3, No. 3, 307-314 (1972) Selective Polymerization of 2-Isopropenyl-2-oxazoline and Cross-linking Reaction of the Polymers Tsutomu KAGIYA and Takehisa MATSUDA Department of Hydrocarbon Chemistry, Faculty of Engineering, Kyoto University, Yoshida, Kyoto, Japan. (Received July 17, 1971) ABSTRACT: The selective polymerization of 2-isopropenyl-2-oxazoline possessing two polymerizable sites in molecule was studied. Due to a marked difference in .polymeriz­ ability between the isopropenyl and oxazoline groups, this monomer provided soluble linear polymers by radical and cationic polymerization. The former was poly(2-isopro­ penyl-2-oxazoline) with the oxazoline group as a side chain, and the latter N-meth­ acryloyl polyethylenimine. The cross-linking reactions of reactive pendant groups in both polymers, which resulted from the radical and cationic polymerizations, gave cross-linked, insoluble products. KEY WORDS Difunctional Monomer / 2-Isopropenyl-2-oxazoline / Selective Polymerization/ Poly(2-isopropenyl-2-oxazoline) / N-Meth­ acryloyl Polyethylenimine / Cross-linking Reaction / Gelation / 2-Isopropenyl-2-oxazoline is a difunctional for 15 hours according to the literature,8 and monomer possessing an isopropenyl group and purified by careful distillation under reduced oxazoline heterocycle as the polymerizable sites pressure [bp 50.5°C (17.5 mm)]. The identifica­ in molecule. It is reported by T. Kagiya, et al.,1 tion was carried out by the infrared and NMR and several other investigators2- 5 that 2-oxa­ spectra of the product. zolines can be polymerized by cationic catalysts Adipic acid as a cross-linker and a,a'-azobis­ such as stannic chloride or boron trifluoride isobutyronitrile (AIBN), which were commercial­ etherate to give N-acyl or N-aryloyl polyethyl­ ly available products (G.
    [Show full text]
  • Simple Modifications of Branched PEI Lead to Highly Efficient Sirna
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE 1448 Bioconjugate Chem. 2008, 19, 1448–1455 provided by Universität München: Elektronischen Publikationen Simple Modifications of Branched PEI Lead to Highly Efficient siRNA Carriers with Low Toxicity Arkadi Zintchenko,* Alexander Philipp, Ali Dehshahri, and Ernst Wagner Center for Drug Research, Department of Pharmacy, Pharmaceutical Biology-Biotechnology, and Center for NanoScience (CeNS), Ludwig-Maximilians-University, Butenandstr. 5-13, D-81377 Munich, Germany. Received February 19, 2008; Revised Manuscript Received May 7, 2008 Polymer carriers like PEI which proved their efficiency in DNA delivery were found to be far less effective for the applications with siRNA. In the current study, we generated a number of nontoxic derivates of branched PEI through modification of amines by ethyl acrylate, acetylation of primary amines, or introduction of negatively charged propionic acid or succinic acid groups to the polymer structure. The resulting products showed high efficiency in siRNA-mediated knockdown of target gene. In particular, succinylation of branched PEI resulted in up to 10-fold lower polymer toxicity in comparison to unmodified PEI. Formulations of siRNA with succinylated PEI were able to induce remarkable knockdown (80% relative to untreated cells) of target luciferase gene at the lowest tested siRNA concentration of 50 nM in Neuro2ALuc cells. The polyplex stability assay revealed that the efficiency of formulations which are stable in physiological saline is independent of the affinity of siRNA to the polymer chain. The improved properties of modified PEI as siRNA carrier are largely a consequence of the lower polymer toxicity.
    [Show full text]
  • Optimization Study of Polyethyleneimine Surface Coating for Microtip DNA Purification
    Optimization Study of Polyethyleneimine Surface Coating for Microtip DNA Purification Andrew Cairns A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science University of Washington 2013 Committee: Jae-Hyun Chung Amy Shen Kyong-Hoon Lee Program Authorized to Offer Degree: Mechanical Engineering ©Copyright 2013 Andrew Cairns University of Washington Abstract Optimization Study of Polyethyleneimine Surface Coating for Microtip DNA Purification Andrew Cairns Chair of the Supervisory Committee: Associate Professor Jae-Hyun Chung Mechanical Engineering DNA purification is a process to separate and extract DNA from various samples. As such, it is an initial step in molecular diagnostics, forensics, pharmacogenomics, and many other genetic analysis based disciplines. Current commercial solutions employ microfilters, centrifugation, or magnetic beads to extract DNA. Recently, silicon microtips have shown equivalent yield to the aforementioned methods, while using fewer reagents and shorter processing time. The working principle of the microtip system relies on dielectrophoretic force and capillary action to concentrate and bind DNA to the microtip surface. There remain two challenges associated with the microtip method: (1) increase efficacy of the process in the presence of a high concentration of inhibitors, such as in blood and (2) improve stability of the microtip surface chemistry during storage. In this thesis, we investigate a polymer film of Polyethyleneimine (PEI) as a potential surface layer to address these challenges. To evaluate the performance of PEI, a coating is applied to the microtips, and their purification performance is tested in a variety of conditions. The operation parameters including curing temperature, capture time, elution time, and elution temperature are optimized.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2010/0197888 A1 Adib Et Al
    US 2010O197888A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0197888 A1 Adib et al. (43) Pub. Date: Aug. 5, 2010 (54) METHOD FOR MANUFACTURING LINEAR Related U.S. Application Data POLYETHYLENIMINE (PED FOR TRANSFECTION PURPOSE AND LINEAR PE (60) Provisional application No. 60/952,993, filed on Jul. OBTANED WITH SUCH METHOD 31, 2007. Publication Classification (75) Inventors: Abdennaji Adib, Illkirch (FR): (51) Int. Cl. Fabrice Stock, Lingolsheim (FR): C08G 73/06 (2006.01) Patrick Erbacher, Illkirch (52) U.S. Cl. ........................................................ 528/423 Graffenstaden (FR) (57) ABSTRACT Correspondence Address: The invention concerns a method of synthesising and prepar BANNER & WITCOFF, LTD. ing linear polyethylenimine (PEI) for use as a transfection 1100 13th STREET, N.W., SUITE 1200 vector, and the product obtained with such a method. It com prises drying a monomer 2-ethyl-2-oxazoline and polymeris WASHINGTON, DC 20005-4051 (US) ing said monomer for obtaining poly (2-ethyl-2-oxazoline) (PEOX) by: using acetonitrile as solvent, adding a dried ini (73) Assignee: Polyplus Transfection, Ilkirch tiator of the reaction of polymerisation, and mixing them Graffenstaden (FR) altogether, purifying said obtained PEOX by evaporation, while performing at least three times successive washing/ (21) Appl. No.: 12/671,312 precipitation steps with methanol and diethyl ether and cor responding filtrations, in order to obtain (i), by performing (22) PCT Fled: Jul. 31, 2008 1H-NMR tests, correct identification of said PEOX polymer, confirmation of absence of monomer to a level <1.0% and (86) PCT NO.: PCT/B2O08/OO2339 confirmation of absence of solvent to a level <5.0% and (ii), by performing Gel Permeation Chromatography, a mean of S371 (c)(1), molecular weight (Mw)>23,000 Da and polydispersity (Mw/ (2), (4) Date: Apr.
    [Show full text]
  • SPECIFIC and EFFICIENT in VIVO DELIVERY of DNA and Sirna by POLYETHYLENIMINE and ITS DERIVATIVES
    SPECIFIC AND EFFICIENT IN VIVO DELIVERY OF DNA AND siRNA BY POLYETHYLENIMINE AND ITS DERIVATIVES by MASSACHUSETTS INSTITUTE OF TECHNOLOGY JENNIFER A. FORTUNE SEP 2 22010 B.A. Chemistry Wheaton College, 2003 LIBRARIES Submitted to the Department of Chemistry in Partial Fulfillment of the Requirements for the Degree of ARCHVES Doctor of Philosophy in Biological Chemistry at the MASSACHUSETTS INSTITUTE OF TECHNOLOGY September 2010 @ 2010 Massachusetts Institute of Technology All rights reserved Signature of Author:. Department of Chemistry August 3, 2010 Certified by: Alexander M. Klibanov Novartis ChairProfessor of Chemistry and Bioengineering Thesis Supervisor Accepted by:. Robert W. Field Haslam and Dewey Professor of Chemistry Chairman,Departmental Committee on Graduate Students This Doctoral Thesis has been examined by a committee of the Department of Chemistry as follows: JoAnne Stubbe Novartis Professorof Chemistry and Professor of Biology Thesis Chair Alexander M. Klibanov Novartis Chair Professor of Chemistry and Bioengineering Thesis Supervisor John M.Essigmann William R. and Betsy P. Leitch Professor of Chemistry andiologicalEngineering SPECIFIC AND EFFICIENT IN VIVO DELIVERY OF DNA AND siRNA BY POLYETHYLENIMINE AND ITS DERIVATIVES by JENNIFER A. FORTUNE Submitted to the Department of Chemistry on August 3, 2010 in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in Biological Chemistry ABSTRACT Linear polyethylenimine (PEI) is the "gold standard" of polycationic gene delivery vectors. However, little focus has been placed on enhancing or understanding the specificity of PEI- mediated gene delivery. Herein we evaluated the effect of chemical modification on the specificity of PEI-mediated nucleic acid delivery. We found that low molecular weight PEI (2 kDa) does not mediate efficient gene expression while high molecular weight (> 87 kDa) leads to toxicity.
    [Show full text]
  • Temperature and Ph Responsive Polyethylenimine Systems As Potential Nonviral Gene Vectors
    Wright State University CORE Scholar Browse all Theses and Dissertations Theses and Dissertations 2010 Temperature and pH Responsive Polyethylenimine Systems as Potential Nonviral Gene Vectors S. Chad Skidmore Wright State University Follow this and additional works at: https://corescholar.libraries.wright.edu/etd_all Part of the Chemistry Commons Repository Citation Skidmore, S. Chad, "Temperature and pH Responsive Polyethylenimine Systems as Potential Nonviral Gene Vectors" (2010). Browse all Theses and Dissertations. 1011. https://corescholar.libraries.wright.edu/etd_all/1011 This Thesis is brought to you for free and open access by the Theses and Dissertations at CORE Scholar. It has been accepted for inclusion in Browse all Theses and Dissertations by an authorized administrator of CORE Scholar. For more information, please contact [email protected]. TEMPERATURE AND pH RESPONSIVE POLYETHYLENIMINE SYSTEMS AS POTENTIAL NONVIRAL GENE VECTORS A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science By S. CHAD SKIDMORE B.A., Case Western Reserve University, 2008 2010 Wright State University School of Graduate Studies July 26, 2010 I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPPERVISION BY S. Chad Skidmore ENTITLED Temperature and pH Responsive Polyethylenimine Systems as Potential Nonviral Gene Vectors BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science. _______________________________ Eric Fossum, Ph.D. Thesis Advisor _______________________________ Kenneth Turnbull, Ph.D. Department Chair Committee on Final Examination _______________________________ Eric Fossum, Ph.D. _______________________________ Kenneth Turnbull, Ph.D. _______________________________ Ioana Pavel, Ph.D. _______________________________ Andrew T. Hsu, Ph.D. Dean, School of Graduate Studies Abstract Skidmore, S.
    [Show full text]
  • Nanoparticles of Quaternary Ammonium Polyethylenimine Derivatives for Application in Dental Materials
    polymers Review Nanoparticles of Quaternary Ammonium Polyethylenimine Derivatives for Application in Dental Materials Marta Chrószcz * and Izabela Barszczewska-Rybarek Department of Physical Chemistry and Technology of Polymers, Silesian University of Technology, 44-100 Gliwice, Poland; [email protected] * Correspondence: [email protected]; Tel.: +48-(32)-2371793 Received: 9 September 2020; Accepted: 28 October 2020; Published: 30 October 2020 Abstract: Various quaternary ammonium polyethylenimine (QA-PEI) derivatives have been synthesized in order to obtain nanoparticles. Due to their antibacterial activity and non-toxicity towards mammalian cells, the QA-PEI nanoparticles have been tested extensively regarding potential applications as biocidal additives in various dental composite materials. Their impact has been examined mostly for dimethacrylate-based restorative materials; however, dental cements, root canal pastes, and orthodontic adhesives have also been tested. Results of those studies showed that the addition of small quantities of QA-PEI nanoparticles, from 0.5 to 2 wt.%, led to efficient and long-lasting antibacterial effects. However, it was also discovered that the intensity of the biocidal activity strongly depended on several chemical factors, including the degree of crosslinking, length of alkyl telomeric chains, degree of N-alkylation, degree of N-methylation, counterion type, and pH. Importantly, the presence of QA-PEI nanoparticles in the studied dental composites did not negatively impact the degree of conversion in the composite matrix, nor its mechanical properties. In this review, we summarized these features and functions in order to present QA-PEI nanoparticles as modern and promising additives for dental materials that can impart unique antibacterial characteristics without deteriorating the products’ structures or mechanical properties.
    [Show full text]
  • Starch Xanthate-Polyethylenimine Reaction Mechanisms
    3693 JOCR:-AL OF POLY:\IER SCIE:-CE, Polymer Chemistry Edition YOL. 13. 1-HI-1-156 iIQ75l Starch Xanthate-Polyethylenimine Reaction Mechanisms :'If. E. CARB, B. T. HOFRElTER, and C. H.. H.USSELL, Northern Re(liollal Research Laboratory, Auricultural Research Service, C.S. Department of Ayriculture. Peoria, Illinois 6'16'04- Synopsis Thl' C'hemiC'al nature of starch xanthate (SX)-polyethylenimine (PEl) reaction produC'ts ha.~ been studied berause of their effertiveness as wet-end additives for im­ prllving strength properties of paper. :\lodel rompounds, in ronjunction with ultra­ vitllct, infrared, and chemical analyses, served to elucidate SX-PEl reaction mecha­ nisms. Aqueous solutions of ~X (degree:s of :sub:;titution O.l-D.;,) were titrated with PEl at pI! :.-i (2,i-:lQoC) to form SX-PEl !locrulent precipitate:s that were determined til hl' polyelectrolyte complexes. However. when :solutions of SX-PEl were kept at pH Ill-I:!. produc·ts were formed that induded dithiocarbamic acid salts in major quantities, PEl thiourea.-, and minor quantities of O-:starch PEl thionrarbamate. Acid precipita­ tion of thesl' ~X-PEI polymeriC' reaction products from their alkaline solutions, which ('Ilntained residual xanthate and PEl. also yielded polyelectrolyte complexe:;. :\lodel ,.'·-tems suggest th:lt PEl thiurum disulfide and starch xanthide, pos:;ible products of air lOxidal iOII, ('ould he present in minor amount:; and would react rapidly with PEl to yield thi'l\lreas and thioncarbamates, respectively. Apparently, mixtures of xanthate and alnin!' gave (1) dithioearbamic aeid salts from both xanthate groups and CS, (decom­ positilln from xanthate), (2) thioureas from both dithioearbamic aeid salts and thiuram eli,ulfide, and (;l) thionearbamates, principally from xanthate a:s opposed to xanthide.
    [Show full text]
  • Full Deacylation of Polyethylenimine Dramatically Boosts Its Gene Delivery Efficiency and Specificity to Mouse Lung
    Full deacylation of polyethylenimine dramatically boosts its gene delivery efficiency and specificity to mouse lung Mini Thomas*, James J. Lu†‡, Qing Ge†, Chengcheng Zhang§, Jianzhu Chen†, and Alexander M. Klibanov*¶ *Department of Chemistry and Division of Biological Engineering, †Center for Cancer Research and Department of Biology, ‡Department of Chemical Engineering, and §Whitehead Institute of Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA 02139 Contributed by Alexander M. Klibanov, March 12, 2005 High-molecular-mass polyethylenimines (PEIs) are widely used ethylenimines (PEIs) (14, 15), are inferior to their viral coun- vectors for nucleic acid delivery. We found that removal of the terparts in the efficiency and specificity of nucleic acid delivery. residual N-acyl moieties from commercial linear 25-kDa PEI en- In the present study, we show that commercial preparations of hanced its plasmid DNA delivery efficiency 21 times in vitro, as well linear PEI (molecular mass ϭ 25 kDa) contain residual N-acyl as 10,000 times in mice with a concomitant 1,500-fold enhance- groups, which severely handicap nucleic acid transfection. Re- ment in lung specificity. Several additional linear PEIs were syn- moval of the residual N-acyl groups dramatically enhances DNA thesized by acid-catalyzed hydrolysis of poly(2-ethyl-2-oxazoline), transfection efficiency of this PEI. Consistent with these find- yielding the pure polycations. PEI87 and PEI217 exhibited the ings, three additional linear PEIs, synthesized ab initio in the highest efficiency in vitro: 115-fold and 6-fold above those of the nonacylated form, exhibited remarkable DNA and short inter- commercial and deacylated PEI25s, respectively; moreover, PEI87 fering RNA (siRNA) delivery efficiency and specificity.
    [Show full text]