Latent Cysteine Residues on Polymers prepared via Free and Controlled Radical Polymerizations

Douglas Amato

Part I
Latent cysteine residues formed via RAFT

Goals of Click Chemistry

Quantitative yields Tolerant to functional groups Insensitive to solvents Example:
Copper mediated azide alkyne cycloaddition
R N N R
3

+ R' N 3

Cu(I)

N R'

Development of New Click Rxns

Variety of Click reactions currently exist
Diels-Alder (DA) Thiol-ene/yne (TE/TY) Oxime

Limitations
Require an external stimulus (DA & TE/TY) Require additional reagents (TE/TY) Goal - Develop Click chemistry that is biocompatible, nonreversible, and does not require an external stimulus to add new functionality to existing materials
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Thiazolidine linkages
Reaction is simple

No external stimulus or additional reagents

Not thermally reversible

Example of thiazolidine linkages

Grinstaff lab at Boston University Utilized as suturing material in eye surgery Challenging sythesis Limited through-put
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Reversible AdditionFragmentation Chain Transfer (RAFT) Polymerization

Predetermined Mn Low PDI Able to prepare block copolymers

RAFT
Wide range of monomers Terminated with transfer agent Can reduce transfer agent to thiol functionality

Part of thiazolidine linkage Need amine functionality

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Thiazolidine Prepolymer
Prepare a styrene-block-acrylonitrile copolymer

Reduce to form a cysteine like functionality

Only need/want a few AN repeat units

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Polymer Synthesis

Simple and efficient synthesis

Able to prepare large quantities of material

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Step-Wise Reduction Method

Reduce and prepare disulfide to prevent coupling

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Coupling Experiments

Clean shift in GPC indicates simple and effective coupling

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Limitations and Solutions

LAH : effective but incompatible with other functional groups
Use tetrabutylammonium borohydride (TBABH4)

New transfer agents

Prepare easy to synthesize RAFT transfer agents

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New Transfer Agents

Simple and efficient synthesis

Able to polymerize styrenes and acrylates

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TA:1 Polymer Synthesis

Modification of previously established method
New AN extension conditions

Conditions : A) TA-1, RAFT; B) AN, RAFT; C) MTS, propyl amine; D) i TBABH4, THF, EtOH ii - DTT, isophthaldehyde

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Polystyrene

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PS-PAN

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PS-PAN-S-S-CH3

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Coupling

TA-1 and TA-3 showed best coupling, but not near ideal. TA-1 used for optimization due ease of synthesis
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Potential Issues
Solvent Reaction time Dithiothreitol Basic solution

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Solvent / Reaction Time

More polar = higher coupling

Time for thiazolidine ring formation insignificant
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Dithiothreitol

More DTT = higher coupling efficiency

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Potassium Carbonate

More K2CO3 = Increase in coupling

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Conclusions / Future Work

Trithiocarbonates unable to achieve 100 % coupling Coupling conditions were attempted to be optimized
Dependent upon : [K2CO3], [DTT], solvent

New goal: synthesis of a single molecule that contains cysteine residue to attach to polymer

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Part II
Latent small molecule cysteine residues

Goals
1. Design a molecule that contains a latent (i.e. protected) cysteine residue

2. Incorporate molecule into a polymer

3. Prove exposure of cysteine residues

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Initial synthetic scheme

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 Synthesis of mesylated compound successful. However required column to purify

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Mesylate to thioacetate

Showed multiple spots via TLC Alternative route investigated

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New Approach
Start with cysteine and add protecting groups

Thia

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1H

NMR
d

b a c Thia b c a d

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Next step

Forma-thia
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1H

NMR

thia

Forma thia

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Alkylation of carboxyl group

Ester proved unstable in acid

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Solution
Use acid stable linkage such as amides

Various carbodiimides did not prove to be effective

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Alternative to carbodiimide coupling

Mixed anhydride

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Proof of amide formation

formathia

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Incorporation into Polymeric System

Need to synthesize polymerizable amines.

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Routes
Synthesis of 4-vinylbenzyl amine:

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Synthesis of 4-vinylbenzyl amine

a

a a

a a

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Ligate polymerizable amine

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NMR

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Polymerization of protected molecule

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NMR

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Issues with deprotection

IR could not detect difference from protected vs. unprotected Need higher amount of cysteine within the backbone

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Alternative synthesis
First attach propargyl amine

Then click azide to alkyne

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NMR

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Evidence of reversibility

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Conclusion
Latent cysteine residues derived from post-polymerization modification are possible, yet are not fully optimized. A facile, high yielding set of reactions have been found to produce a protected cysteine

The deprotection of the cysteine has been shown

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Future Work
Attach alkyne protected cysteine to polymers with pendant azides Thiolene/yne chemistry with latent thiol residue in RAFT Creation of new materials with pendent cysteines.

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Acknowledgements
Advisor: Dr. Philip Costanzo Thesis Committee:
Prof. Chad Immoos

Prof. Derek Gragson

Funding provided by an award from the Research Corporation for Science Advancement
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Costanzo Research Group

Dahlia Amato Anton Chavez Julia Dean Guilhem Dehoe Thaddeus Formal Alex London Tristan Kleine

Craig Machado
Miles Markmann Leanna Monteleone Chris Pattillo Dimitri Pappademos Alicia Ross Kim Varney Anthony Varni
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Polymers & Coatings Program

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Chemistry & Biochemistry Department

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Thank You

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