Applications of vinyl sulfones in the field of bioconjugations. The vinyl sulfone is a chemical function with "click-chemistry" characteristic that enable it for a rapid and efficient covalent linkage of the compounds derivatized with this function and those bearing complementay chemical functions (thiol and amines)

1. glycohcembio.ugr.es
R S
O
O

2. Javier
Lopez-­‐Jaramillo
/ljara@ugr.es
Francisco
Santoyo-­‐Gonzalez
fsantoyo@ugr.es
Permanet
Members
Fernando
Hernandez-­‐Mateo
Amateo@ugr.es

3. Applied
Science
Basic
Science
Scien6fic
Ac6vity

4. SO2
R S
O
O
Research
Lines
N
N
N
Synthesis
of
Neoglycoconjugates
Cyclodextrins
&
Glycocyclodextrins
Carbon
Nanopar6cles
1,3-­‐Dipolar
Cycloaddi6ons
Cyclic
Sulfates
Gene
Transfec6on
Polymeric
matrix
from
polysaccharides

5. “Chemistry
tailored
to
generate
substances
quickly
and
reliably
by
joining
units
together”
H.
C.
Kolb,
M.
G.
Finn,
K.
B.
Sharpless,
Angew.
Chem.
Inter.
Ed.
2001,
40,
2004-­‐2021
The
click-­‐chemistry
philosophy
Modular
and
wide
in
scope
High
chemical
yield
Inoffensive
byproducts
Readily
available
star>ng
materials
and
reagents
Stereospecific
Simple
reac>on
condi>ons
No
solvent
or
benign
solvents
H2O
Easy
product
isola>on.
Not
chromatography
Physiologically
stable
High
atom
economy
Click-­‐Chemistry

6. Click-­‐chemistry
Reac>ons
Cycloaddi>ons
of
unsaturated
species
1,3-­‐Dipolar
cycloaddi>on
reac>ons
Nucleophilic
subs>tu>on
chemistry
Ring-­‐opening
reac>ons
of
strained
heterocyclic
electrophiles
Epoxides
Aziridines
Aziridinium
ions
Episulfonium
ions
Non-­‐aldol
type
carbonyl
chemistry
Ureas
Thioureas
Aroma>c
heterocycles
Oxime
Ethers
Hydrazones
Amides
Addi>ons
to
C-­‐C
mul>ple
bonds
Epoxida>on
Dihydroxyla>on
Aziridina>on
Sulfenyl
halide
addi>on
Michael
addi>ons
of
Nu-­‐H
reactants
Click-­‐Chemistry

7. Azide-­‐Alkyne
Cycloaddi6on
(ACC)
N
N
NR
R'
R
R' N3
1-­‐3
Dipolar
cycloaddi6ons
N
N
N
Cu
(I)
catalyzed
click-­‐chemistry
(CuAAC)
Metal-­‐free
click-­‐chemistry
“Click-­‐Chemistry”

8. (Ph3P)3·∙CuBr
(EtO)3P·∙CuI
N
N
NR
R'Cu(I)
R R' N3
Cu(I)
complex
as
organic
soluble
catalyst
for
CuAAC
MW
MW
Org.
Le4,
2000,
2,
2499
Org.
Le4,
2003,
5,
1951
cited
by
240
N
N
N
1-­‐3
Dipolar
cycloaddi6ons
Featured
contribu6ons
in
CuAAC
CarboUGR

9.
Glyco
clusters
(Homo
and
Hetero)
Glyco
dendrons
Glyco
dendrimers
Glyco
polymers
Glyco
Cyclodextrins
Glyco-­‐silicas
Bio>n-­‐silica
Func>onal
Glycoconjugates
N
N
N
1-­‐3
Dipolar
cycloaddi6ons
Featured
applica6ons
of
CuAAC
CarboUGR

10.
N
N
NR
R'
Catalysis
Methodology
Applications
Chem.
Soc.
Rev.,
2009,
38,
3449
Tetrahedron.,
2005,
61,
9338
Org.
Biomol.
Chem.,
2007,
2291
Adv.
Synth.
Catal.,
2006,
348,
2410
Adv.
Synth.
Catal.,
2010,
352,
3306
Adv.
Synth.
Catal.,
2012,
en
prensa
Org.
Le4,
2000,
2,
2499
Org.
Le4,
2003,
5,
1951
J.
Org.
Chem.,
2008,
73,
7768
J.
Org.
Chem.,
2008,
73,
7772
Eur.
J.
Org.
Chem.,
2009,
2441
Eur.
J.
Org.
Chem.,
2009,
2454
N
N
N
1-­‐3
Dipolar
cycloaddi6ons
Featured
Publica6ons
CarboUGR

11. Non-­‐AAC
Click-­‐reac6ons
“Thiol-­‐Ene
”
R SH
R'
R
S
R'
R SH
R'
“Thiol-­‐Yne
”
Catalyst
Catalyst
“Michael-­‐Type
Addi6on
”
(1,4-­‐
addi6on
or
conjugate
addi6on)
R
S
R'
S
R
R''
H
R'R
acceptor
donor
R
R''
R'
H
R,
R’
and
R’’
electron-­‐withdrawing
groups
Base
B:
Click-­‐Chemistry
R S
O
O
Vinyl
Sulfones

12. Conjugated
addi6ons
ü Efficiency
ü High
yields
ü No
sub-­‐products
Reac6vity
R S
O
O
Vinyl
Sulfones
R S
O
O
R
S
O O
NuR
S
O O
Nu-+
Nu- = C-, N-, y S-Nucleófilos
Cycloaddi6on
reac6ons
ü [2+2]
Cycloaddi6ons
ü [3+2]
Cycloaddi6ons
ü [4+2]
Cycloaddi6ons
(Diels-­‐Alder
Mixed
reac6ons
ü Epoxida6on
ü Reduc6ons
ü Methatesis
with
olefines
ü Asimetrid
dyhydroxila6ons

13. Bioconjuga6on
Labelling
Cross-­‐linkers
Catalysis
Immobiliza6on
R S
O
O
R S
O
O
Vinyl
Sulfones
Func6onal
Labelling

14. Non-­‐Magnetic
and
Magnetic
heterogeneous
catalyst
Cu(I)
Scavengers
N
N
N
R1
R2
R1
R2 N3
+
Lm Cu+
Lm1) Cu(I), 2)
CuAAC
Lm
Lm Cu+
Cu+
N
N
N
R1
R2
R1
R2 N3
+
Lm Cu+
Lm1) Cu(I), 2)
CuAAC
Lm
Lm Cu+
Cu+
Cu(I)
Sorbents
based
in
soported
chelating
agents
ü Robust
ü Efficient
ü Low
doses
ü Negligible
Cu
Leaching
ü Simple
operational
protocols:
Filtration
or
magnetic
decantation
ü Recyclability
ü High
efficiency
ü Isolation
of
metal-­‐free
clicked
compounds
LmLm Cu+
Heterogeneous
Catalyst
Scavengers
Catalysis
R S
O
O
Vinyl
Sulfones
N N
N
n n n = 0, 1
N N
NH
N
H
N
S
O2
LmLm
R
SiO2
O2
S
H
S
O2
LmNH2 Lm
R= NH2R= SO2CH=CH2
Lm

15. Removal
of
heavy
metals
α,β-­‐
CD
Homogeneous
Matrix
Heterogeneous
matrix
Starch
Dextrin
Cationic
matrix
Anionic
matrix
Cr3+,
Pb2+
y
Cd2+
Removal
of
organic
dyes
Removal
of
organic
contaminants:
Fenols
S
O
O
N+
HO
H2N
OH
O
H2N
OH
O
HO
O
N
H
OH
O
OH
O
HS
CO3
-­‐2
,
0,5
M,
pH
12
Polymeric
adsorbents
based
on
polysaccharides
and
cyclodextrins
for
water
purification
ES
2334756
A1
R S
O
O
Vinyl
Sulfones
Cross-­‐linkers

16. R S
O
O
Vinyl
Sulfones
Bioconjuga6on
R S
O
O
From
Chemical
reac6vity
to
Bioconjuga6on
R
S
O O
NuR
S
O O Nu-
Nu- = C-, N-, y S-Nucleófilosü Efficiency
ü High
yields
ü No
sub-­‐products
ü Physiological
conditions
-­‐
Aqueous
media,
-­‐
Mild
alkaline
pH,
-­‐
Room
temperature
ü No
catalyst
and/or
subproducts
Nu-­‐
=
Lys,
Cys,
His

17. Bioconjuga6on
Labelling
Immobiliza6on
R S
O
O
Vinyl
Sulfones
Func6onal
Labelling
R S
O
O
Glycosyda6on
Drug
delivery
Lipida6on
Sensors
Glycoarrays
Hybrid
materials
Fluorophores
Redox
Bio6n

18. Bioconjuga6on
Labelling
R S
O
O
Vinyl
Sulfones
R S
O
O
Fluorophores
Redox
Bio6n

19. Biotine
Fluorophore
Fluorescent
Labelling
S
X
O
O
X
R
S
O2
O
S
O2
2
ON N
N
O
+
Rhodamina B VS
N
SO O
R
O2
S
O
O2
S
2
N
Dansilo VS
S
NHHN
HH
O
O
3
R
S
O2
O
S
O2
2
N
Biotina VS
S
X
O
O
S
O
O
S
O
O
O2
S
O
2
VS
S
NHHN
HH
O
O
3
R
S
O2
O
S
O2
2
N
Biotina VS
Biotinylation
Redox
labelling
S
O
O
S
O
O
Fe
S
X
O
O
Fe
S
O
O
X
=
NH2,
SH
ConA,
BSA,
lisozime,
Avidine
Fe
O S
O
O
Fe
S
O
O
R S
O
O
Vinyl
Sulfones
Labelling
Labelling
Reagents
S
O
O

20. Bio6n
Fluorophore
HRP
AVST
BTSAP BTSAP
Ruta a
Ruta b
Ruta b
Bifunctional
labelling
Triazolyl
containing
vinyl
sulfones
as
double-­‐labeling
agents
and
their
preparation
and
use
in
the
marking
of
biomolecules
WO
2009/106665
A1
(PCT
Int.
Appl.)
R S
O
O
Vinyl
Sulfones
Route
A
Route
B

21. Bioconjuga6on
R S
O
O
Vinyl
Sulfones
Func6onal
Labelling
R S
O
O
Glycosyla6on
Drug
delivery
Lipida6on

22. S
O
O
S
S
O
O
S
Neoglicoproteins
Glycosyla6on
BSA,
ConA,
Lisozyme
Lumazine,
Apoferritin
VS
azúcares
O S
S
O
O
O SH
HO n
AcO n
α-D-Man, β-D-Glc, β-D-GlcNAc, α-L-Fuc
β-Lac, β-Mal, β-Mel
R S
O
O
Vinyl
Sulfones
Func6onal
labelling:
Glycosyla6on
Carbohydrate-­‐func6onalized
Magne6c
Nanopar6cles
Maghemite
(γ-­‐Fe2O3)
nanopar6cles
Apoferritin
pH-­‐Driven
templated
assembly
(Encapsula6on)
Glycosyla6on

23. ü High
efficiency
ü Low
toxicity
ü Maintenance
of
properties
in
the
presence
of
serum
ü Different
cell
lines
Improved
transfection
efficiency
(NH2)16
(NH2)16-n
R1S
O
O
NH
n
(NH2)16-n
S
O
O
NH
n
R1
O
NH
Y
(NH2)16-n
S
O
O
NH
n
O
R1 S
O
O
NH
(NH2)15.5-n
S
O
O
NH
0.5n
R1 S
O
O
PAMAN G2
Rho
PAMAM-­‐G2-­‐based
amphiphilic
non-­‐viral
gene
vectors
R1 S
O
O
R1 = (CH2)2Me, (CH2)10Me, (CH2)16Me,
O
S
O2
R1
O
N
H
S
O
O
Y
Y = O, S
z
(CH2)7CH=CH(CH2)7Me
O
N
ON+ N
S
O2
O
S
O2
R
+
2
Rhodamine
B
VS
Rho
R S
O
O
Vinyl
Sulfones
Func6onal
labelling:
Lipida6on

24. Preparation
of
lipids
with
vinylsulfonyl
funcionality
for
therapeutic
use
in
the
transport
of
therapeuticals
ES
2337226
A1
Targeted
delivery
of
ISCOM
Inmunolabelling
essay
for
ISCOM
targe6ng
delivery
to
Trypanosoma
cruzi
R S
O
O
Vinyl
Sulfones
Func6onal
labelling:
Drug
delivery
CO3
-­‐2
buffer
0,125
M
pH
8,
12
h,
4º
C
ISCOM-­‐Ficocianine
ISCOM-­‐
drug

25. Mono
VS-­‐Cyclodextrin
R S
O
O
Vinyl
Sulfones
Func6onal
labelling:
Drug
delivery
O
OH
HO
N
O
O
OH
HO
OH
O
O
OH
OH
OH
O
O
OH
OH
OH
OO
OH
OH
HO
O
O OH
OH
HO
O
OTs
O
OH
HO
HO
O
S
O2
HOS
O
O
S
O
O
O2
S
Conjuga6on
Nanobody
O
N
H
N
H
N OH
O
O2N
Nitrofurazone
Encapsula6on
S
O
O
O2
S
Therapeu6c
func6on
Carrier
func6on
Targe6ng
func6on
Tripanosoma
Cruzi
ü Payload
flexibility
ü Target
flexibility
by
adequate
selection
of
the
recognizable
biomolecule
ü Multifunctional
decoupling:
therapeutic,
carrier
and
targeting
Modular
design
Poly-­‐His

26. Mono
VS-­‐Cyclodextrin
R S
O
O
Vinyl
Sulfones
Func6onal
labelling:
Drug
delivery
S
O
O
Conjuga6on
Protein
A
Encapsula6on
Therapeu6c
func6on
Carrier
func6on
Targe6ng
func6on
Tripanosoma
Cruzi
S
O
O
O2
S
O
N
O
OHO
N
Camptotecine
(Cpt)
S
O
O
O2
S
5-­‐chloro-­‐1H-­‐benzo
[d]imidazole-­‐2-­‐thiol
N
H
N
SH
Cl
S
O
O
O2
S
Modular
design
ü Payload
flexibility
ü Target
flexibility
by
adequate
selection
of
the
IgG
ü Multifunctional
decoupling:
therapeutic,
carrier
and
targeting
IgG

27. Bioconjuga6on
Immobiliza6on
R S
O
O
Vinyl
Sulfones
R S
O
O
Sensors
Glycoarrays
Hybrid
materials

28. PROTEOMIC
“Ad-­‐hoc”
chromatographic
supports
INDUSTRIAL
PROCESS
Immobilization
of
Enzymes
FUNCTIONAL
HYBRID
MATERIALS
VS
func6onalized
silica
X
SiO2
O
O
O
O
O
X
Y = NMe, S
Y3Si
S
Y= NMe, Y = S
SiO2
O
O
O
Si Y3
S
O
O
X
=
Lys,
Cys,
His
ü Symplicity
ü Efficiency
ü Green
chemistry
Silica-­‐vinylsulfone
compound,
synthesis
and
uses
as
immobilization
surface
WO
2009/040460
A1
(PCT
Int.
Appl.)
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Hybrid
materials
A”Ready
to
Use”
pre-­‐ac6vated
material
S
O
O
SiO2

29. VS
func6onalized
silica
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Hybrid
materials
SiO2
O
O
O
Si S
O
O
O
N
H
SH
O
H
N
O
OH
O
HO
NH2
Glutathione
(GSH)
h-­‐Type
Thioredoxins
u Silica-­‐glutathione
for
the
isolation
of
proteins
fused
with
GST
u Pull-­‐down
experiments
for
proteins
interacting
whit
thioredoxine
“Ad-­‐hoc”
chromatographic
supports
S
O
O
SiO2

30. VS
func6onalized
silica
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Hybrid
materials
S
O
O
S
O
O
S
O
O
S
O
O Apoferri6n
Lactoferrin
Immobiliza6on
An6microbial
ac6vity
against
E.
Coli
High
Fe-­‐Uptake
capacity
Immobilization
of
Enzymes
S
O
O
SiO2
Functional
Hybrid
Materials
X
X
=
Lys,
Cys,
His
S
O
O
SiO2 S
O
O
SiO2
X
Invertase,
Lactase,
Lysozyme

31. Super-­‐paramagne6c
Hybrid
Nanopar6cles
Polymer
Compounds
Having
Immobilizing
Properties
WO
2012/085303
A1
(PCT
Int.
Appl.)
O
O
N
HSS
O
O
O
N
Me
O OH
H2N
Invertase,
Avidin
HRP
Vinyl
sulfone
SP-­‐NPs
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Hybrid
materials

32. S
O
O S
S
O
O
S
HN
LECTIN
COMMERCIALLY
LABELLED
LECTIN
LECTIN
COMERCIALLY
LABELLED
LECTIN
S
O
O
S
HN
S
O
O
S
Glyco-­‐array
Immobiliza6on
DETECTION
ISOLATION
VS
Sugars
NH2 NH2
NH2 NH2
NH2 NH2
NH2NH2
placas de
micropozillos
placas de vidrio
placas de vidrio
injertadas con polisina
S
O
O S
S
O
O
S
HN
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Hybrid
materials
Glass
slides
Microwell
plates
Polylysine
coated
glass
slides

33. O
NH
S
HN
H
N
H H
3
O
R
R S
O
O
Vinyl
Sulfones
Immobiliza6on:
Sensors
R
Bio6nylated
pyrrols
NH2 O
NH
NH
O
Luminol
R
N
S
O2
N
S
O2
H
N
Bio6nyla6on
Electropolymeriza6on
Electrochemiluminiscence
bio6nylated
polymers
Electropolymeriza6on
S
O2
2
N
SO O
R
O2
S
O
O2
S
2
N
Dansilo VS
S
NHHN
HH
O
O
3
R
S
O2
O
S
O2
2
N
Biotina VS
S
O
O
Bio6nyla6on
Bio6nylated
Cholesterol
Oxidase
HO Cholesterol
Electrochemiluminiscent
disposable
cholesterol
biosensor
Avidin
Gold

34. Acta
Cryst.,
2005,
F61,
435
J.
Org.
Chem.,
2010,
75,
4039
J.
Mat.
Chem.,
2010,
20,
7189
Org.
Biomol.
Chem,
2013,
11,
2586
Org.
Biomol.
Chem.,
2011,
9,
851
Org.
Biomol.
Chem.,
2010,
8,
7189
Org.
Biomol.
Chem.,
2010,
8,
667
Bioconjugate
Chem.,
2012,
846
Scien6fic
Produc6on
R S
O
O
Vinyl
Sulfones
ChemMedChem.,
2014,
9,
383
2013,
13,
61
Int.
J.
Nanomed.,
2012,
7,
5491
Anal,
Chim.
Acta.,
2012,
754,
91
J.
Am.
Chem.
Soc.,
2011,
133,
4889
Adv.
Synth.
Catal.,
2010,
3306
J.
Plant
Physiol.,
2010,
423

35. R S
O
O
Knowledge
Transfer
Triazolyl
containing
vinyl
sulfones
as
double-­‐labeling
agents
and
their
preparation
and
use
in
the
marking
of
biomolecules
WO
2009/106665
A1
(PCT
Int.
Appl.)
Single-­‐labeling
agents
based
on
vinyl
sulfones
and
their
preparation
and
use
in
marking
biomolecules
such
as
protein
WO
2009/106664
A1
(PCT
Int.
Appl.)
Compound
for
labeling
biomolecules
based
on
vinyl
sulfone,
their
preparation
and
use
in
marking
biomolecules
such
as
proteins
WO/2009144344
A2
(PCT
Int.
Appl.)
Silica-­‐vinylsulfone
compound,
synthesis
and
uses
as
immobilization
surface
WO
2009/040460
A1
(PCT
Int.
Appl.)
Polymeric
adsorbents
based
on
polysaccharides
and
cyclodextrins
for
water
purification
ES
2334756
A1
Drug
delivery
and
transfection
agents
based
on
alkylsulfonate
functionalized
PAMAM
dendrimers
ES
2351909
A1
R S
O
O
Vinyl
Sulfones

36. R S
O
O
Simple
and
efficient
chemistry
R S
O
O
Vinyl
Sulfones
for
smart
applica6ons

37. Collabora6ons
CTS-­‐183
Bioquímica
y
Parasitología
Molecular
A.
Osuna
Carillo
Parasitología
J.
F.
Corpas
Bioquímica,
Biología
Celular
y
Molecular
de
Plantas
FQM-­‐118
Solid
Phase
Spectrometry
Group
Luis
F.
Capitan
Valley
Química
Analí6ca
FQM-­‐368
BionNanoMetal
José
M.
Domínguez
Vera
Química
Inorgánica
CVI-­‐212
Regulación
Bioquímica
y
Gené6ca
Rafael
Salto
González
Bioquímica
y
Biología
Molecular
II
BIO-­‐286
Bioquimica
y
Señalizacion
Celular
J.
B.
Barroso-­‐Albarran
Bioquimica
y
Biologia
Celular
Former
Ph
fellows
• Alicia
Megia-­‐Fernández
• Julia
Morales-­‐SanFrutos
• Francisco
Pérez
Balderas
CarboUGR
Non-­‐Permanent
Members
• Mariano
Ortega-­‐Muñoz
R S
O
O

38. Research
Lines