glycohcembio.ugr.es
R SO
O
Javier Lopez-‐Jaramillo /[email protected]
Francisco Santoyo-‐Gonzalez [email protected]
Permanet Members
Fernando Hernandez-‐Mateo [email protected]
Applied Science
Basic Science
Scien6fic Ac6vity
SO2
R SO
O
Research Lines
NN
N
Synthesis of Neoglycoconjugates
Cyclodextrins & Glycocyclodextrins
Carbon Nanopar6cles
1,3-‐Dipolar Cycloaddi6ons
Cyclic Sulfates
Gene Transfec6on
Polymeric matrix from polysaccharides
“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
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
Azide-‐Alkyne Cycloaddi6on (ACC)
NN
NR
R'
R
R' N3
1-‐3 Dipolar cycloaddi6ons NN
N
Cu (I) catalyzed click-‐chemistry
(CuAAC)
Metal-‐free click-‐chemistry
“Click-‐Chemistry”
(Ph3P)3·∙CuBr (EtO)3P·∙CuI
NN
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
NN
N 1-‐3 Dipolar cycloaddi6ons
Featured contribu6ons in CuAAC CarboUGR
Glyco clusters (Homo and Hetero)
Glyco dendrons
Glyco dendrimers
Glyco polymers
Glyco Cyclodextrins
Glyco-‐silicas Bio>n-‐silica
Func>onal Glycoconjugates
NN
N 1-‐3 Dipolar cycloaddi6ons Featured applica6ons of CuAAC CarboUGR
NN
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
NN
N 1-‐3 Dipolar cycloaddi6ons
Featured Publica6ons
CarboUGR
Non-‐AAC Click-‐reac6ons
“Thiol-‐Ene ”
R SH
R'
RS
R'
R SH
R'
“Thiol-‐Yne ”
Catalyst Catalyst
“Michael-‐Type Addi6on ” (1,4-‐ addi6on or conjugate addi6on)
RS
R'
SR
R''
H
R'R
acceptor
donor
R
R''
R'
H
R, R’ and R’’ electron-‐withdrawing groups Base B:
Click-‐Chemistry
R SO
O
Vinyl Sulfones
Conjugated addi6ons
ü Efficiency ü High yields ü No sub-‐products
Reac6vity
R SO
OVinyl Sulfones
R SO
O
RS
O O
NuRS
O ONu-+
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
Bioconjuga6on Labelling
Cross-‐linkers Catalysis
Immobiliza6on
R SO
O
R SO
OVinyl Sulfones
Func6onal Labelling
Non-‐Magnetic and Magnetic heterogeneous catalyst
Cu(I) Scavengers
NN
NR1
R2
R1
R2 N3
+
Lm Cu+
Lm1) Cu(I), 2)
CuAAC
Lm
Lm Cu+
Cu+ NN
NR1
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 SO
OVinyl Sulfones
N N
Nn n n = 0, 1N N
NH
NH
NSO2
LmLm
R
SiO2O2S
H
SO2
LmNH2 LmR= NH2R= SO2CH=CH2
Lm
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 OO
N+HO
H2NOH
OH2N
OH
O
HO
ONH
OH
O
OH
OHS
CO3-‐2 , 0,5 M, pH 12
Polymeric adsorbents based on polysaccharides and cyclodextrins for water purification ES 2334756 A1
R SO
OVinyl Sulfones
Cross-‐linkers
R SO
OVinyl Sulfones Bioconjuga6on
R SO
O
From Chemical reac6vity to Bioconjuga6on
RS
O O
NuRS
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
Bioconjuga6on
Labelling Immobiliza6on
R SO
OVinyl Sulfones
Func6onal Labelling
R SO
O
Glycosyda6on Drug delivery
Lipida6on
Sensors
Glycoarrays
Hybrid materials
Fluorophores
Redox
Bio6n
Bioconjuga6on
Labelling
R SO
OVinyl Sulfones
R SO
O
Fluorophores
Redox
Bio6n
Biotine Fluorophore
Fluorescent Labelling
SX
O
O
X
R
SO2
OSO2
2
ON N
N
O
+
Rhodamina B VS
N
SO O
RO2S
O
O2S
2N
Dansilo VS
S
NHHNHH
O
O3
R
SO2
OSO2
2N
Biotina VS
SX
O
O
SO
OSO
O
R
SO2
OSO2
2
ON N
N
O
+
Rhodamina B VS
N
SO O
RO2S
O
O2S
2N
Dansilo VS
S
NHHNHH
O
O3
R
SO2
OSO2
2N
Biotina VS
Biotinylation Redox labelling SO
O
SO
O
FeSX
O
O
Fe
S
O
O
X = NH2, SH ConA, BSA, lisozime, Avidine
Fe
O SO
O
Fe
S
O
O
R SO
OVinyl Sulfones Labelling
Labelling Reagents
SO
O
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 SO
OVinyl Sulfones
Route A
Route B
Bioconjuga6on
R SO
OVinyl Sulfones
Func6onal Labelling
R SO
O
Glycosyla6on Drug delivery
Lipida6on
SO
OSS
OOS
Neoglicoproteins
Glycosyla6on
BSA, ConA, Lisozyme Lumazine, Apoferritin VS azúcares
O S SO
O
O SH
HO nAcO n
α-D-Man, β-D-Glc, β-D-GlcNAc, α-L-Fucβ-Lac, β-Mal, β-Mel
R SO
OVinyl Sulfones
Func6onal labelling: Glycosyla6on
Carbohydrate-‐func6onalized Magne6c Nanopar6cles
Maghemite (γ-‐Fe2O3) nanopar6cles
Apoferritin
pH-‐Driven templated assembly (Encapsula6on) Glycosyla6on
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
R1SO
O
NHn
(NH2)16-n
SO
O
NH
n
R1
O
NHY
(NH2)16-n
SO
O
NH
n
O
R1 SO
O
NH
(NH2)15.5-n
SO
O
NH0.5n
R1 SO
O
PAMAN G2
Rho
PAMAM-‐G2-‐based amphiphilic non-‐viral gene vectors
R1 SO
O
R1 = (CH2)2Me, (CH2)10Me, (CH2)16Me,
O SO2
R1
O
NH SO
O
Y
Y = O, S
z(CH2)7CH=CH(CH2)7Me
O
N
ON+ N
SO2
O SO2
R
+
2
Rhodamine B VS
Rho
R SO
OVinyl Sulfones Func6onal labelling: Lipida6on
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 SO
OVinyl Sulfones Func6onal labelling: Drug delivery
CO3-‐2 buffer 0,125 M
pH 8, 12 h, 4º C
ISCOM-‐Ficocianine ISCOM-‐ drug
Mono VS-‐Cyclodextrin
R SO
OVinyl Sulfones Func6onal labelling: Drug delivery
O
OHHO
N
O
OOH
HOOHO
OOH
OH
OH
O
O
OHOH
OH
OO
OH
OH
HOO
O OH
OHHO
O
OTs
OOH
HO
HO
O
SO2
HOSO
O
SO
O
O2S
Conjuga6on
Nanobody
ONHN
HN OH
OO2N
Nitrofurazone
Encapsula6on
SO
O
O2S
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
Mono VS-‐Cyclodextrin R S
O
OVinyl Sulfones Func6onal labelling: Drug delivery
SO
OConjuga6on
Protein A
Encapsula6on
Therapeu6c func6on
Carrier func6on
Targe6ng func6on
Tripanosoma Cruzi
SO
O
O2S
ON
O
OHON
Camptotecine (Cpt)
SO
O
O2S
5-‐chloro-‐1H-‐benzo [d]imidazole-‐2-‐thiol
N
HN
SHCl
SO
O
O2S
Modular design
ü Payload flexibility ü Target flexibility by adequate selection of the IgG ü Multifunctional decoupling: therapeutic, carrier and targeting
IgG
Bioconjuga6on
Immobiliza6on
R SO
OVinyl Sulfones
R SO
O
Sensors
Glycoarrays
Hybrid materials
PROTEOMIC “Ad-‐hoc” chromatographic supports
INDUSTRIAL PROCESS Immobilization of Enzymes
FUNCTIONAL HYBRID MATERIALS
VS func6onalized silica
X
SiO2
OOO
O
O
X
Y = NMe, S
Y3Si S
Y= NMe, Y = S
SiO2
OO
OSi Y3
SO
OX = 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 SO
OVinyl Sulfones Immobiliza6on: Hybrid materials
A”Ready to Use” pre-‐ac6vated material
SO
OSiO2
VS func6onalized silica
R SO
OVinyl Sulfones Immobiliza6on: Hybrid materials
SiO2OOO
Si SO
OO
NH
SH
O
HN
O
OH
O
HONH2
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
SO
OSiO2
VS func6onalized silica
R SO
OVinyl Sulfones Immobiliza6on: Hybrid materials
SO
O SO
OS
OO
SO
O Apoferri6n Lactoferrin Immobiliza6on
An6microbial ac6vity against E. Coli
High Fe-‐Uptake capacity
Immobilization of Enzymes
SO
OSiO2
Functional Hybrid Materials
X
X = Lys, Cys, His SO
OSiO2 S
O
OSiO2 X
Invertase, Lactase, Lysozyme
Super-‐paramagne6c Hybrid Nanopar6cles
Polymer Compounds Having Immobilizing Properties WO 2012/085303 A1 (PCT Int. Appl.)
O
O
NHSS
O
O
O N
Me
O OH
H2N
Invertase, Avidin HRP
Vinyl sulfone SP-‐NPs
R SO
OVinyl Sulfones Immobiliza6on: Hybrid materials
SO
O S
S
OOS
HN
LECTIN COMMERCIALLY LABELLED LECTIN LECTIN COMERCIALLY
LABELLED LECTIN
S
OOS
HN
S
OOS
Glyco-‐array
Immobiliza6on
DETECTION
ISOLATION
VS Sugars
NH2 NH2
NH2 NH2
NH2 NH2NH2NH2placas de
micropozillos
placas de vidrio
placas de vidrioinjertadas con polisina
SO
O S
S
OOS
HN
R SO
OVinyl Sulfones Immobiliza6on: Hybrid materials
Glass slides
Microwell plates
Polylysine coated glass slides
O
NH
S
HNHN
H H
3O
R
R SO
OVinyl Sulfones Immobiliza6on: Sensors
R
Bio6nylated pyrrols
NH2 O
NHNH
O
Luminol
R
NSO2
NSO2
HN
Bio6nyla6on Electropolymeriza6on
Electrochemiluminiscence bio6nylated polymers
Electropolymeriza6on
R
SO2
OSO2
2
ON N
N
O
+
Rhodamina B VS
N
SO O
RO2S
O
O2S
2N
Dansilo VS
S
NHHNHH
O
O3
R
SO2
OSO2
2N
Biotina VS
SO
O
Bio6nyla6on
Bio6nylated Cholesterol Oxidase
HO Cholesterol
Electrochemiluminiscent disposable cholesterol biosensor
Avidin
Gold
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
OVinyl 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
R SO
OKnowledge 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 SO
OVinyl Sulfones
R SO
O
Simple and efficient chemistry
R SO
OVinyl Sulfones
for smart applica6ons
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 SO
O
Research Lines