UNIT OF GLYCOCONJUGATE CHEMISTRY
Group Leader: Gemma Arsequell
The aim of the Unit is to study biochemical or medicinal chemistry issues by using chemical methodologies. Most frequently used tools are peptide and carbohydrate chemistry, halogenation reactions, aqueous organometallic catalysis and proteomic techniques. Traditional fields of interest are enzyme catalysis, pain and immunity related mechanisms, transthyretin amyloidosis inhibitors and more recently, Alzheimer’s disease (AD) interfering compounds.
Members of the Unit are usually involved in multidisciplinary projects involving computer scientists, biochemical, biological, pharmacological, conformational (NMR), crystallographic and nuclear chemistry groups at national and international level.
Pain and immunity related compounds. Natural glycoconjugates and glycoconjugation
Studies on antinociceptive compounds. One main aim of the Unit has been the search for new drug candidates for either severe or moderate pain and related mechanisms. Thanks to a project funded by Fundació La Marató 2006, we aimed first to intervene in the metabolic pathway of enkephalin degradation and started the study of a pain-related molecule called opiorphin, a dual inhibitor of the ectopeptidases neprilysin (NEP) and aminopeptidase N (APN) that metabolizes a number of neuropeptides among them the enkephalins. Opiorphin produces analgesia in humans by inhibiting the enkephalin degradation. On the other hand, we looked at one of the natural metabolites of morphine, the glycoconjugate known as M6G, which was under clinical development for the treatment of pain. During this project we synthesized and tested new analogues of both opiorphin, M6G and other opioid peptides to gain new insights for the discovery of new candidates for analgesic drugs. Research on this field is the result of a long-standing interdisciplinary collaboration with two important research groups in Spain. One of them with expertise in pain research, is lead by Prof. Raquel E. Rodriguez, Director of Instituto de Neurociencias de Castilla y León (INCyL) in Salamanca and the other one with expertise in Structural Biology (NMR studies) lead by Prof. Jesús Jiménez-Barbero at CIC BioGUNE in Derio (Vizcaya)..
Glycopeptides and Glycoimmunology
Both CD4+ and CD8+ T cells can recognize glycopeptides carrying mono- and disaccharides in a MHC-restricted manner provided the glycan group is attached to the peptide at suitable positions. In such glycopeptides, the primed T cells recognize the glycan structure. The question of T cell recognition of glycopeptides may be important in the immune defense against microorganisms, because many microbial antigens are in fact glycosylated. T cell recognition of glycans may also play an important role in the immune defence against tumors.
Drug discovery in Alzheimer disease: Small molecule chaperones of the interaction of transthyretin and Abeta peptides as drug candidates for Alzheimer’s disease.
Protein-protein interaction modulation by small molecules to identify AD interfering compounds.
These studies are an extension of the drug discovery effort initiated in year 2000 to find drug candidates for a group of rare diseases associated to transthyretin (TTR) which is a thyroid hormone transporter protein. These systemic amyloid diseases are always triggered by single point hereditary or spontaneous mutations on the protein. The pharmacological intervention we have been pursuing relies in small molecule compounds that resemble thyroid hormones and attach to the binding pocked of TTR and thus stabilize its tetrameric structure by preventing its dissociation and further misfolding and aggregation of its monomers into amyloid fibrils and deposits. In the course of this research we found that a particular set of TTR stabilizing compounds enhance the TTR-Aβ interaction when studied in vitro. This prompted us to study the activity of one of such compounds, namely Iododiflunisal (IDIF), when administered in an AD animal model. By using an AβPPswe/PS1A246E model and orally administering the drug we found that the drug is able to stabilize TTR in the plasma of these animals. Also, it was evidenced that the drug penetrated into the brain. The drug decreased Aβ levels and Aβ deposition in the brain. Cognitive functions associated with AD-like neuropathologies were also ameliorated in these animals.
Figure: A) Small molecule TTR/Aβ chaperone Iododiflunisal (IDIF) developed in our group; B) TTR structure where the surface pocket, the retinol binding protein (RBP)-binding pocket, and the T4 central channel binding pocket are indicated; C) TTR-Aβ-iododiflunisal model obtained through molecular docking
This work was conducted in association with our long standing partners at the IBMC of Porto (Portugal). These in vivo studies prompted us to settle a drug discovery program focused in the discovery of small-molecule compounds enhancers of the transthyretin-Abeta interaction that may lead to potential AD modulating drugs. The project received a grant from Fundació La Marató de TV3 (Neurodegenerative diseases, 2013): “Setting a rational screening program for transthyretin-Abeta binding stabilizing compounds that may lead to potential Alzheimer´s disease modulating drugs”. The consortia is integrated by five multidisciplinary research teams with tracked expertise in different disciplines and with a translational vision. The project gathers experts in computational chemistry, medicinal chemistry/radiochemistry, structural and cell biology, molecular neurobiology of AD and molecular imaging.
By structural and computational studies we have found that Abeta(12−28) is the main recognition element of the Abeta peptide in the interaction with TTR. The NMR results, assisted by molecular modeling protocols, have provided the first structural model for the TTR-Abeta interaction, as well as for the ternary complex formed in the presence of IDIF. These results have been published in J. Med. Chem. 2017, 60, 5749-5758).
Figure: NMR studies of the interaction of TTR and Abeta peptides
http://www.ccma.cat/tv3/marato/es/projectes-financats/2013/212/
Drug discovery in Rare Diseases: Transthyretin related amyloid rare diseases.
Chemical methodologies: The Barluenga’s reagent as a tool in biochemistry. Effects of halogenation on biological properties of active compounds.
- Binding of common organic UV-filters to the thyroid hormone transport protein transthyretin using in vitro and in silico studies: Potential implications in health.
Ellen Y. Cotrina, Angela Oliveira, Jordi Llop, Jordi Quintana, Xevi Biarnés, Isabel Cardoso, M. Silvia Díaz-Cruz, Gemma Arsequell
Environmental Research, Volume 217, 15 January 2023, 114836
- Longitudinal evaluation of neuroinflammation and oxidative stress in a mouse model of Alzheimer disease using positron emission tomography
Rejc, L., Gómez-Vallejo, V., Joya, A., Arsequell, G., Egimendia, A., Castellnou, P., Ríos-Anglada, X., Cossío, U., Baz, Z., Iglesias, L., Capetillo-Zarate, E., Ramos Cabrer, P., Martin, A., Llop, J.
Alzheimer's Research and Therapy, 14 (1), art. no. 80, 2022
- Static discrete disorder in the crystal structure of iododiflunisal: on the importance of hydrogen bond, halogen bond and π-stacking interactions
Barbas, R., Font-Bardia, M., Ballesteros, A., Arsequell, G., Prohens, R., Frontera, A.
CrystEngComm, 2022
- Targeting transthyretin in Alzheimer's disease: Drug discovery of small-molecule chaperones as disease-modifying drug candidates for Alzheimer's disease
Cotrina, E.Y., Santos, L.M., Rivas, J., Blasi, D., Leite, J.P., Liz, M.A., Busquets, M.A., Planas, A., Prohens, R., Gimeno, A., Jiménez-Barbero, J., Gales, L., Llop, J., Quintana, J., Cardoso, I., Arsequell, G.
European Journal of Medicinal Chemistry, 226, art. no. 113847, 2021
- Neuroprotection in early stages of Alzheimer’s disease is promoted by transthyretin angiogenic properties
Gião, T., Saavedra, J., Vieira, J.R., Pinto, M.T., Arsequell, G., Cardoso, I.
Alzheimer's Research and Therapy, 13 (1), art. no. 143, 2021
- Does Antigen Glycosylation Impact the HIV-Specific T Cell Immunity?
Olvera, A., Cedeño, S., Llano, A., Mothe, B., Sanchez, J., Arsequell, G., Brander, C.
Frontiers in Immunology, 11, art. no. 573928, .2021
- Preparative scale production of recombinant human transthyretin for biophysical studies of protein-ligand and protein-protein interactions
Cotrina, E.Y., Vilà, M., Nieto, J., Arsequell, G., Planas, A.
International Journal of Molecular Sciences, 21 (24), art. no. 9640, pp. 1-12. 2020
- Optimization of kinetic stabilizers of tetrameric transthyretin: A prospective ligand efficiency-guided approach
Cotrina, E.Y., Blasi, D., Vilà, M., Planas, A., Abad-Zapatero, C., Centeno, N.B., Quintana, J., Arsequell, G.
Bioorganic and Medicinal Chemistry, 28 (23), art. no. 115794, 2020
- Repurposing benzbromarone for familial amyloid polyneuropathy: A new transthyretin tetramer stabilizer
Cotrina, E.Y., Oliveira, Â., Leite, J.P., Llop, J., Gales, L., Quintana, J., Cardoso, I., Arsequell, G.
International Journal of Molecular Sciences, 21 (19), art. no. 7166, pp. 1-16. 2020
- An Assay for Screening Potential Drug Candidates for Alzheimer's Disease That Act as Chaperones of the Transthyretin and Amyloid-β Peptides Interaction
Cotrina, E.Y., Gimeno, A., Llop, J., Jiménez-Barbero, J., Quintana, J., Prohens, R., Cardoso, I., Arsequell, G.
Chemistry - A European Journal, 26 (72), pp. 17462-17469. 2020
- Radiochemical examination of transthyretin (TTR) brain penetration assisted by iododiflunisal, a TTR tetramer stabilizer and a new candidate drug for AD
Rios, X., Gómez-Vallejo, V., Martín, A., Cossío, U., Morcillo, M.Á., Alemi, M., Cardoso, I., Quintana, J., Jiménez-Barbero, J., Cotrina, E.Y., Valencia, G., Arsequell, G., Llop, J.
Scientific reports, 9 (1), p. 13672. 2019
- Atorvastatin in PLGA-PEG Nanoparticles Derivatized with the HIV-TAT Peptide Protects Neuronal Cultures in an Oxygen-Glucose Deprivation (OGD) Model
Angel Cespedes, Maria Jose Perez, Maria Jose Gomara, Francisco Wandosell, Isabel Haro
Model. Am J Nanotechnol Nanomed. 2018; 1(2): 055-063.
- Benzyl-2-Acetamido-2-Deoxy-α-d-Galactopyranoside Increases Human Immunodeficiency Virus Replication and Viral Outgrowth Efficacy In Vitro.
Olvera A, Martinez JP, Casadellà M, Llano A, Rosás M, Mothe B, Ruiz-Riol M, Arsequell G, Valencia G, Noguera-Julian M, Paredes R, Meyerhans A, Brander C.
Front Immunol. 2018, 26; 8:2010.
- Insights on the Interaction between Transthyretin and Aβ in Solution. A Saturation Transfer Difference (STD) NMR Analysis of the Role of Iododiflunisal.
Gimeno A.; Santos, L. M.; Alemi, M.; Rivas, J.; Blasi, D.; Cotrina, .Y.; Llop, J.; Valencia, G.; Cardoso, I.; Quintana, J.; Arsequell, G.; Jiménez-Barbero, J.
- Med. Chem. 2017, 60, 5749-5758.
- Role of the sugar moiety on the opioid receptor binding and conformation of a series of enkephalin neoglycopeptides.
Rosa M, Gonzalez-Nunez V, Barreto-Valer K, Marcelo F, Sánchez-Sánchez J, Calle LP, Arévalo JC, Rodríguez RE, Jiménez-Barbero J, Arsequell G, Valencia G.
Bioorg Med Chem. 2017, 25, 2260-2265.
- Tuning transthyretin amyloidosis inhibition properties of iododiflunisal by combinatorial engineering of the nonsalicylic ring substitutions.
Vilaró M, Nieto J, La Parra JR, Almeida MR, Ballesteros A, Planas A, Arsequell G, Valencia G.
ACS Comb. Sci. 2015, 17, 32-38.
- Optimized Proteomic Mass Spectrometry Characterization of Recombinant Human μ-Opioid Receptor Functionally Expressed in Pichia pastoris Cell Lines.
Rosa M, Bech-Serra JJ, Canals F, Zajac JM, Talmont F, Arsequell G, Valencia G
- Proteome Res. 2015, 14, 3162-3173.
- Influence of polar side chains modifications on the dual enkephalinase inhibitory activity and conformation of human opiorphin, a pain perception related peptide.
Rosa M, Marcelo F, Calle LP, Rougeot C, Jiménez-Barbero J, Arsequell G, Valencia G.
Bioorg. Med. Chem. Lett. 2015, 25, 5190-5193.
- Transthyretin Stabilization by Iododiflunisal Promotes Amyloid- Peptide Clearance, Decreases its Deposition, and Ameliorates Cognitive Deficits in an Alzheimer's Disease Mouse Model.
Ribeiro CA, Oliveira SM, Guido LF, Magalhães A, Valencia G, Arsequell G, Saraiva MJ, Cardoso I.
J. Alzheimer´s Dis. 2014, 39, 357-370.
- Modulation of the Fibrillogenesis Inhibition Properties of Two Transthyretin Ligands by Halogenation
Cotrina EY, Pinto M, Bosch L, Vilà M, Blasi D, Quintana J, Centeno NB, Arsequell G, Planas A, Valencia G.
J. Med. Chem. 2013, 56, 9110-9121.
- Methods to Evaluate the Inhibition of TTR Fibrillogenesis Induced by Small Ligands.
Arsequell G, Planas A.
Curr. Med. Chem. 2012, 19, 2343-2355.
- Structure-activity relationship study of opiorphin, a human dual ectopeptidase inhibitor with antinociceptive properties.
Rosa M, Arsequell G, Rougeot C, Calle LP, Marcelo F, Pinto M, Centeno NB, Jiménez-Barbero J, Valencia G.
J. Med. Chem. 2012, 55, 1181-1188.
