Click Chemistry

Presented by:
Eman Jamil El-agroudy

Submitted to:
Prof. Dr/ Omaima Abo-Elwafa

Click chemistry
Click chemistry is a chemical philosophy introduced by K. Barry Sharpless , in 2001 and describes chemistry tailored to generate substances quickly and reliably by joining small units together. This is inspired by the fact that nature also generates substances by joining small modular units. Click chemistry is not a specific reaction; it is a concept that mimics nature.

Advantages of click chemistry labeling:

1-Labeling by simple, efficient reactions 2-High labeling yield (nearly quantitative) 3-Allows introduction of sensitive labels (mild conditions) 4-Highly modular system (combinatorial approach) 5-Orthogonal to conventional methods 6-Bioorthogonal reaction (no azides or alkynes in natural systems) 7-Labeling of all bases possible (G, A, C, T, U) 8-Broad range of marker- and dye-azides available 9-Also adaptable to linking beads, microarrays, nanoparticles, peptides etc. 10-generate only inoffensive byproducts 11-be stereospecific 12-be physiologically stable 13-exhibit a large thermodynamic driving force > 84 kJ/mol to favor a reaction with a single reaction product. A distinct exothermic reaction makes a reactant "spring-loaded". 14-have high atom economy.

Requirments:
1-have simple reaction conditions 2-use readily available starting materials and reagents 3-use no solvent or use a solvent that is benign or easily removed (preferably water) 4-provide simple product isolation by non-chromatographic methods (crystallisation or distillation)

Applications of click chemistry:

1-preparative organic synthesis of 1,4-substituted triazoles 2-modification of natural products and pharmaceuticals 3-drug discovery 4-macrocyclizations using Cu(I) catalyzed triazole couplings 5-Protein and peptides conjugation to polymers and surfaces using oxime chemistry 6-Click chemistry in supramolecular materials, calixarenes, rotaxanes, and catenanes 7-carbohydrate clusters and carbohydrate conjugation by Cu(1) catalyzed triazole ligation reactions 8-Dendrimer synthesis and functionalization by click chemistry for biomedical application 9-Reversible Diels- Alder cycloaddition for the design of multifunctional network polymers 10-Click chemistry in preparation of biohybrid materials 11-Functional nanomaterials using the Cu- catalyzed Huisgen Cycloaddition Reaction 12-Copper catalyzed click chemistry for surface engineering 13-Click chemistry in protein engineering, Design, Detection and Profiling 14-material science 15-The role of click chemistry in polymer synthesis 16-Flourogenic Cu(I)- catalyzed Azide Alkyne Cycloaddition Reactions and their applications in Bioconjugation. for example, azidocoumarin 17-Synthesis and functionalization of biomolecules via click chemistry

Also Click Chemistry represents an easy to use technology and has enhanced research in fields such as:
DNA and RNA labeling incorporation of alkyne-phosphoramidites in oligos and labeling with marker azides (single- or multi-labeling) PCR assays, PCR primers and labeling of large fragments use of alkyne-triphosphates in the nucleotide mixture and labeling of the resulting PCR-fragments with marker azides, or use of oligonucleotides as multi-labeled primers for pre- or post-synthetic modification FISH probes and FISH experiments use of alkyne-modified oligonucleotides and labeling with marker azides (pre- or post-hybridization) PEGylation introduction of PEG-tags via click chemistry with outstanding yields and modularity Microarrays use of phosphoramidites, triphosphates or oligonucleotides to set up microarrays

Some chemical reactions that fall within the field of click chemistry:

Explaination of some important applications of click chemistry: A-Cu catalyzed click reactions Alkyne-containing Reagents for Cu(I)-catalyzed Click Reactions
The Copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) or Cu(I)-catalyzed 1,3dipolar Huisgen cycloaddition of alkynes and azides ,is the most prominent example of click reactions, it uses Copper (Cu) as a catalyst at room temperature. This reaction proceeds with great efficiency and selectivity in aqueous media and yields a triazole moiety.

The use of this method for DNA modification has been somewhat delayed by the fact that copper ions damage DNA, typically yielding strand breaks. As these problems have now been overcome by the use of copper (I) stabilizing ligands (e.g. tris(benzyltriazolylmethyl)amine, TBTA, Carell et al. and Seela et al. discovered that the CuAAC reaction can be used to functionalize alkyne-modified DNA nucleobases with extremely high efficiency.

B-peptides and derivatives:

the click reaction has been used to synthesize diverse derivatives of bioactive compounds via substitution of the originally appearing functionality with the resulting triazole unit, in order to improve the activity of the compounds. Gopi et al. Investigated the structure- activity correlation of peptide conjugates that act as receptor site antagonists of HIV-1 gp 120. The group synthesized derivatives of the original bioactive peptide on solid phases through the reaction of an immobilized azidemodified proline residue with alkynes containing different side chain.

The introduction of a triazole functioality offers the possibility of producing novel -turn mimics. The click reaction of an alkyne- containing dipeptide and the azide containing dipeptide generates different tetrapeptides in a convergent synthesis

The click reaction can be performed chemoselectively. This approach was used to selectively generate two triazole linkages in a one- pot method. The first reaction is traditionally performed in the presence of a copper(I)- species with a dialkynyl-derivative, which contains one free and one TMS- protected triple bond. The second click reaction is carried out in the presence of Cu(I) and Ag(I), which catalyze the deprotection of the second alkynyl functionality. The starting material 12 for those click reactions is easily prepared through the substiution of -chlorinated peptides 11 with sodium azide.

There are examples of intramolecular approaches that result in cyclic peptides. The intramolecular click reactions were applied to the synthesis of cyclic tetrapeptides these tetrapeptides has the function of tyrosine inhibitors. This derivative can be easily obtained via cyclization through peptide coupling, or through triazole formation in the final step.

C-Peptoids:
As triazole- containing mimics of peptides have been extensively investigated, there are other groups who have synthesized peptoids modified through the click reaction of azides with alkynes. The synthesis of functionalized -peptoid macrocycles occurs through linear synthesis of polyalkyne-containing linear -peptoids and subsequent macrocyclization. This cyclic peptoids 20 were then modified by the addition of azides to produce polytriazolecontaining cycles 21 with different residues on triazole side chain.

D-Peptidic Dendrimers:
They are biologically relevant structures, as channels for drug delivery. The functionalization of these dendrimers enables the modulation of the surface to generate special properties, which are essential for their specific interactions. Click chemistry has been used by several groups to design novel dendrimeric structures. An example is the synthesis of certain polyphenylene dendrimers as follows:

E-Oligonucleotides
the click reaction is one of the most powerful methods of labeling oligonucleotides. It can be performed not only on phosphoramidate- modified oligonucleotides, but also on triple bonds and azides mostly attached through 5 or 3- terminal conjugation of the oligonucleotides. Because the 5- position of pyrimidine and the 7-position of purines lies in the major groove of the B-DNA duplex and have steric freedom for additional functionalities, most of the DNA modifications take place in these positions. Some modified nucleosides and oligonucleotides were synthesized rthrough Cu 1catalyzed azide- alkyne cycloaddition. An example is the cycloaddition of compound 31 and AZT 32 to form the triazole product33.

Schematic representation of the cycloaddition reaction performed on duplex DNA.

F- Copper Free Click Reaction

Many copper free and hence non toxic, alternatives to CuAAC exist which are bioorthogonal, which means noninteracting with biology.encompasses all of the charactaristics of such a chemical reaction. A bioorthogonal ligation is thus a chemical reaction in which two functional groups selectively react with one another to form a covalent linkage in the presence of all of the functionality in biological system.

Examples of Cu free click reactions: 1-Staudinger Ligation

The strain-promoted or Cu(I)-free [2+3] cycloaddition strategy relies on the use of strained cyclooctynes. Their use decreases the activation energy for the cycloaddition click reaction, enabling it to be carried out without the need for catalysis at low temperatures with an efficiency greater than that of the Cu(I)-catalyzed ligation. Our conjugation chemistry is based on the reaction of a dibenzylcyclooctyne (DBCO) linker with an azide linker to form a stable triazole. This click reaction is very fast at room temperature, does not require a cytotoxic Cu(I) catalyst and creates stable triazoles. This unique covalent bond is created when DBCO, incorporated into one type of biomolecule, reacts with an azide linker, incorporated into a second biomolecule.

The strain-promoted Click reaction and the so called Staudinger ligation (phosphine-azide) are competing technologies for chemoselective ligation. Both reactions are chemoselective and do not require copper, so both do not damage biomolecules. However, the rate of Staudinger ligation is about 100fold lower than the rate of the DBCO cycloaddition, which makes the Staudinger ligation hardly useful for studying dynamic biological systems. Only in cases where the speed of ligation is irrelevant, both reactions can be used with about equal efficiency.

2-Condensation of Ketones and Aldehydes with Heteroatom-bound amines:

Ketones and Aldehydes can form reversible imine adducts with many amines found in biological systems, this process is thermodynamically unfavourable in water. The use of hydrazides and aminooxy reagents, often called -effect amines, because the heteroatombound amine is much more nucleophilic than simple amines, shifts the equilibrium to the hydrazone and oxime products.

G-Reversible Diels-Alder Cycloaddition for the Design of Multifunctional Network Polymers

Click chemistries such as the Diels- Alder reaction have been used to prepare many novel and difficult to achieve multifunctional polymer networks. Such a chemistry is popular because it offers high yield and minimal side reactions under mild reaction conditions. Diels-Alder reaction is a cycloaddition of a diene and a dienophile to form a substituted cyclohexane. The reaction is sometimes thermally reversible. This reversibility makes the Diels-Alder reaction particularly desirable for the development of multifunctional polymer networks.

Most of literature has focused on the reaction of furan (diene) and maleimide (dienophile).the reaction of maleimide with cyclopentandione, fulvene, pyrone and anthracene, of benzene with cyclopentandione and of quinine with cyclopentandione have been investigated.

The following are some well known thermoreversible Diels-Alder reactions:

Potential limitations of click chemistry:

In spite of the success of the concept of click chemistry within few years, there are few limitations associated with the concept. As a matter of fact, the CuAAC reaction is still by far the most widely used click reaction. However, copper is known to be toxic demonstrated side effects associated with excessive copper intake include hepatitis, Alzeheimers disease and neurological disorders. For click reactions to be used in contact with living systems, the copper catalyst must be completely removed or alternatives, such as Staudinger ligation Azides, among the prime reactants for Huisgens 1,3- dipolar cycloaddition reaction, are also often associated with potential toxic side effects, and certain azides may bear a very real explosive potential Finally, a more practical limitation is that the supply of clickable starting materials often cannot keep up with the demands of the rapidly emerging application space in materials science and biotechnology.

References:

1-http://books.google.com.eg/books?id=9eoyUOu4L9IC&printsec=frontcover&source= gbs_ViewAPI&redir_esc=y#v=onepage&q&f=false 2-http://www.jenabioscience.com/cms/en/1/browse/1379/?gclid=CJKV3N6pa0CFcRH3godnkP1lg 3-http://www.sigmaaldrich.com/chemistry/chemical-synthesis/technologyspotlights/click.html 4-http://www.jenabioscience.com/images/741d0cd7d0/ Copper_Catalyzed_Click_Reactions.pdf 5-http://en.wikipedia.org/wiki/Dendrimer 6-http://www.jenabioscience.com/images/741d0cd7d0/ Copper_Free_Click_Reactions.pdf 7-http://pubs.acs.org/doi/abs/10.1021/ic8017634 8- click chemistry for biotechnology and material science, Joerg Lahann