SUMMARY OF RESEARCH

Target identification for ParasiticTropical Diseases Drug Discovery.

Our lab is interested in exploiting unique aspects of parasitic protozoa metabolism as drug targets for the discovery of new treatments. Kinetoplastid and apicomplexan parasites comprise a group of protozoans responsible for diseases with a serious impact in human health and the socioeconomic growth of developing countries. We actively participate in international consortia together with pharma industry in order to implement a multidisciplinary approach towards drug discovery. Our main focus is nucleotide metabolism and we have identified several unique druggable enzymes that perform essential functions in kinetoplastids. Many nucleoside analogs are already approved for the treatment of patients with cancer or viral diseases. Of particular interest is pyrimidine metabolism where several enzymes are essential for parasite viability. Thus, inhibition of thymidylate (dTMP) biosynthesis severely diminishes the viability of parasitic protozoa. We have characterized several enzymes specifically involved in the formation of dTTP (dUTPase, thymidine kinase, thymidylate kinase) and have shown that depletion results in decreased deoxythymidine triphosphate (dTTP) levels and the so-called thymineless death. In this process the ratio of deoxyuridine triphosphate (dUTP) versus dTTP in the cellular nucleotide pool has a crucial role. A high dUTP/dTTP ratio leads to uracil misincorporation into DNA, the activation of DNA repair pathways, DNA fragmentation and eventually cell death. We are interested in how all these processes take place in parasitic protozoa. In addition we continue to explore the properties, biological function and druggability of additional enzymes involved in nucleotide metabolism.

Carbohydrate-binding agents for the treatment of trypanosomal diseases.

Another area of interest is the exploitation of carbohydrate-binding agents as antitrypanosomals. This is a new approach based on the use of compounds that bind to parasite surface glycans and that lead to rapid killing of trypanosomes. As an example, pradimicin and its derivatives are non-peptidic carbohydrate-binding agents that adhere to the carbohydrate moiety of the parasite surface glycoproteins inducing parasite lysis. Notably, pradimicin S has good pharmaceutical properties and enables cure of an acute form of sleeping sickness in mice. By inducing resistance in vitro we have established that the composition of the sugars attached to the variant surface glycoproteins are critical to the mode of action of pradimicins and play an important role in infectivity. We are currently exploring the potential of this class of compounds in the treatment of other parasitic diseases.

Phenotype-driven novel methodologies for screening using high throughput formats in antiprotozoan drug discovery.

We also have ongoing collaborations with medicinal chemists and the MEDINA foundation in the area of drug discovery through phenotypic screening. MEDINA has one of the biggest collections in the world of microbial natural products. The collection holds 190,000 strains of filamentous fungi, actinomycetes and bacteria, and has contributed to some of the most important discoveries of recent decades in natural product drug discovery. Together we have established an HTS platform for phenotypic screening of the collection. As a result of screening against Plasmodium and trypanosomes, many novel compounds with antiparasitic activities that include known and novel microbial natural products have been identified.

Role of the human NTP pyrophosphatase DCTPP1 in nucleotide homoeostasis

In order to keep the dNTP pool in balance, the synthesis and degradation of DNA precursors must be precisely regulated. Catabolic activities that convert deoxynucleoside triphosphates into their monophosphate form are involved in this process. Human cells possess an all-α-NTP (nucleoside triphosphate) pyrophosphatase named DCTPP1 which is structurally related to trypanosomal dUTPases. We have performed studies in order to evaluate the role of DCTPP1 in the size and composition of dNTP pools and genetic stability. We are extensively characterizing its properties and have established that DCTPP1 has a central role in the balance of dCTP and the metabolism of deoxycytidine analogues, thus contributing to the preservation of genome integrity and to the mode of action of nucleoside analogues used in cancer chemotherapy.

FUNDING AGENCIES LAST 5 YEARS

- NUCLEOTIDO HIDROLASAS EN LA INFECCION POR TRYPANOSOMA. ELUCIDACION DE SU PAPEL EN LAS INTERACCIONES HUESPED-PARASITO. PROYECTO, PN2022 - PROY I+D GENERACION CONOC. - PID, Ref: PID2022-142971OB-I00, (2023 - 2026).

- Repurposing human kinase inhibitor chemotypes for Neglected Tropical Diseases. PROYECTO, Allergy and Infectious Diseases Research - NIH (EEUU), Ref: 2R01AI114685 (SUBAWARD NUMBER, (2021 - 2026).

- Implantación de una plataforma de cribado de alto rendimiento para el descubrimiento de nuevos compuestos anti-SARS-CoV-2.. PROYECTO, INTRAMURAL - PROYECTOS INTRAMURALES(09), Ref: CSIC-COV19-171, (2020 - 2021).

- PAPEL DE LAS NUCLEOTIDOHIDROLASAS EN LA PRESERVACION DE LA INTEGRIDAD GENOMICA EN TRYPANOSOMA BRUCEI. PROYECTO, PN2019 - Proyectos I+D+i «Retos Investigación», Ref: PID2019-109623RB-I00, (2020 - 2023).

- Implicaciones de las nucleótido hidrolasas DCTPP1 y dUTPasa en estabilidad genómica y función mitocondrial: evaluacióndel potencial terapéutico.. PROYECTO, J.A.- Retos de la sociedad andaluza, Ref: P18-RT-4870, (2020 - 2023).

- Red de investigación colaborativa en enfermedades tropicales RICET. PROYECTO, PN2016 - REDES TEMATICAS INV. COOPERATIVA, Ref: RD16/0027/0014, (2017 - 2021).

- MANTENIMIENTO DE LA HOMEOSTASIS DE NUCLEOTIDOS Y DE LA INTEGRIDAD GENOMICA EN TRYPANOSOMA BRUCEI. PROYECTO, PN2016 - PROY I+D+I - PRG. RETOS DE LA SOCIEDAD, Ref: SAF2016-79957-R, (2016 - 2020).

PUBLICATIONS LAST 5 YEARS

-Frederick Boye Annang; Guiomar Pérez-Moreno; Cristina Bosch-Navarrete; Victor González-Menéndez; Jesús Martín; Thomas A. Mackenzie; Maria C. Ramos; Luis M. Ruiz-Pérez; Olga Genilloud; Dolores González-Pacanowska; Francisca Vicente; Fernando Reyes, Antiparasitic Meroterpenoids Isolated from Memnoniella dichroa CF-080171, Pharmaceutics, 2023, Vol. 15: 2-492, ARTÍCULO, Id:919250

-Vidal, A.E.; Yagüe-Capilla, M.; Martínez-Arribas, B.; García-Caballero, D.; Ruiz-Pérez, L.M.; González-Pacanowska, D., Inosine triphosphate pyrophosphatase from Trypanosoma brucei cleanses cytosolic pools from deaminated nucleotides, Scientific Reports, 2022, Vol. 12: 1-6408, ARTÍCULO, Id:893829

-Annang, F.; Pérez-Moreno, G.; Díaz, C.; González-Menéndez, V.; de Pedro Montejo, N.; del Palacio, J.P.; Sánchez, P.; Tanghe, S.; Rodriguez, A.; Pérez-Victoria, I.; Cantizani, J.; Ruiz-Pérez, L.M.; Genilloud, O.; Reyes, F.; Vicente, F.; González-Pacanowska, D., Preclinical evaluation of strasseriolides A–D, potent antiplasmodial macrolides isolated from Strasseria geniculata CF-247,251, Malaria Journal, 2021, Vol. 20: 1-457, ARTÍCULO, Id:867755

-Klug, D.M.; Mavrogiannaki, E.M.; Forbes, K.C.; Silva, L.; Diaz-Gonzalez, R.; Pérez-Moreno, G.; Ceballos-Pérez, G.; Garcia-Hernández, R.; Bosch-Navarrete, C.; Cordón-Obras, C.; Gómez-Liñán, C.; Saura, A.; Momper, J.D.; Martinez-Martinez, M.S.; Manzano, P.; Syed, A.; El-Sakkary, N.; Caffrey, C.R.; Gamarro, F.; Ruiz-Perez, L.M.; Gonzalez Pacanowska, D.; Ferrins, L.; Navarro, M.; Pollastri, M.P., Lead Optimization of 3,5-Disubstituted-7-Azaindoles for the Treatment of Human African Trypanosomiasis, Journal of Medicinal Chemistry, 2021, Vol. 64: 9404-9430, ARTÍCULO, Id:852597

-Yagüe-Capilla, M.; Castillo-Acosta, V.M.; Bosch-Navarrete, C.; Ruiz-Pérez, L.M.; González-Pacanowska, D., A Mitochondrial Orthologue of the dNTP Triphosphohydrolase SAMHD1 Is Essential and Controls Pyrimidine Homeostasis in Trypanosoma brucei, ACS Infectious Diseases, 2021, Vol. 7: 318-332, ARTÍCULO, Id:843857

-Dana M. Klug; Rosario Diaz-Gonzalez; Travis J. DeLano; Eftychia M. Mavrogiannaki; Melissa J. Buske; Raeann M. Dalton; John K. Fisher; Katherine M. Schneider; Vivian Hilborne; Melanie G. Fritsche; , Quillon J. Simpson; Westley F. Tear; William G. Devine; Guiomar Pérez-Moreno; Gloria Ceballos-Pérez; Raquel García-Hernández; Cristina Bosch-Navarrete; Luis Miguel Ruiz-Pérez; Francisco Gamarro; Dolores González-Pacanowska; Maria Santos Martinez-Martinez; Pilar Manzano-Chinchon; Miguel Navarro; Michael P. Pollastri; Lori Ferrins., Structure-property studies of an imidazoquinoline chemotype with antitrypanosomal activity, RSC Medicinal Chemistry, 2020, Vol. 11: 950-959, ARTÍCULO, Id:840926

-Annang, F.; Perez-Moreno, G.; Gonzalez-Menendez, V.; Lacret, R.; Perez-Victoria, I.; Martín, J.; Cantizani, J.; de Pedro, N.; Choquesillo-Lazarte, D.; Ruiz-Perez, L.M.; Gonzalez-Pacanowska, D.; Genilloud, O.; Vicente, F.; Reyes, F., Strasseriolides A−D, A Family of Antiplasmodial Macrolides Isolated from the Fungus Strasseria geniculata CF-247251, Organic Letters, 2020, Vol. 22: 6709-6713, ARTÍCULO, Id:815952

-Martínez-Arribas, B.; Requena, C.E.; Pérez-Moreno, G.; Ruíz-Pérez, L.M.; Vidal, A.E.; González-Pacanowska, D., DCTPP1 prevents a mutator phenotype through the modulation of dCTP, dTTP and dUTP pools, Cellular and Molecular Life Sciences, 2020, Vol. 77: 1645-1660, ARTÍCULO, Id:804605

-Klug, D.M.; Tschiegg, L.; Diaz, R.; Rojas-Barros, D.; Perez-Moreno, G.; Ceballos, G.; García-Hernández, R.; Martinez-Martinez, M.S.; Manzano, P.; Ruiz, L.M.; Caffrey, C.R.; Gamarro, F.; Pacanowska, D.G.; Ferrins, L.; Navarro, M.; Pollastri, M.P., Hit-to-Lead Optimization of Benzoxazepinoindazoles As Human African Trypanosomiasis Therapeutics, Journal of Medicinal Chemistry, 2020, Vol. 63: 2527-2546, ARTÍCULO, Id:804253

-Tear WF; Bag S; Diaz-Gonzalez R; Ceballos-Pérez G; Rojas-Barros DI; Cordon-Obras C; Pérez-Moreno G; García-Hernández R; Martinez-Martinez MS; Ruiz-Perez LM; Gamarro F; Gonzalez Pacanowska D; Caffrey CR; Ferrins L; Manzano P; Navarro M; Pollastri MP, Selectivity and Physicochemical Optimization of Repurposed Pyrazolo[1,5-b]pyridazines for the Treatment of Human African Trypanosomiasis, Journal of Medicinal Chemistry, 2020, Vol. 63: 756-783, ARTÍCULO, Id:779338

-Pérez-Moreno G; Sánchez-Carrasco P; Ruiz-Pérez LM; Johansson NG; Müller S; Baragaña B; Hampton SE; Gilbert IH; Kaiser M; Sarkar S; Pandurangan T; Kumar V; González-Pacanowska D, Validation of Plasmodium falciparum dUTPase as the target of 5¿-tritylated deoxyuridine analogues with anti-malarial activity, Malaria Journal, 2019, Vol. 18: 392 , ARTÍCULO, Id:773950

-Ana Moro-Bulnes; Víctor M. Castillo-Acosta; Maria Valente; Juana Carrero-Lérida; Guiomar Pérez-Moreno; Luis Miguel Ruiz-Pérez; Dolores González-Pacanowska, Contribution of Cytidine Deaminase to Thymidylate Biosynthesis in Trypanosoma brucei: Intracellular Localization and Properties of the Enzyme, mSphere, 2019, Vol. 4: 4-e00374-19, ARTÍCULO, Id:768900

-Klug DM; Diaz-Gonzalez R; Pérez-Moreno G; Ceballos-Pérez G; García-Hernández R; Gomez-Pérez V; Ruiz-Pérez LM; Rojas-Barros DI; Gamarro F; González-Pacanowska D; Martínez-Martínez MS; Manzano P; Ferrins L; Caffrey CR; Navarro M; Pollastri MP, Evaluation of a class of isatinoids identified from a high-throughput screen of human kinase inhibitors as anti-Sleeping Sickness agents, PLoS Neglected Tropical Diseases, 2019, Vol. 13: e00071292-e0007129, ARTÍCULO, Id:742826

-Yagüe-Capilla M; García-Caballero D; Aguilar-Pereyra F; Castillo-Acosta VM; Ruiz-Pérez LM; Vidal AE; González-Pacanowska D, Base excision repair plays an important role in the protection against nitric oxide-and in vitro-induced DNA damage in Trypanosoma brucei, Free Radical Biology and Medicine, 2019, Vol. 131: 59-71, ARTÍCULO, Id:730212

-Annang F; Pérez-Victoria I; Pérez-Moreno G; Domingo E; González I; Tormo JR; Martín J; Ruiz-Pérez LM; Genilloud O; González-Pacanowska D; Vicente F, Reyes F, MDN-0185, an Antiplasmodial Polycyclic Xanthone Isolated from Micromonospora sp. CA-256353, Journal of Natural Products, 2018, Vol. 81: 1687-1691, ARTÍCULO, Id:722901

-Schiafino-Ortega S; Baglioni E; Pérez-Moreno G; Marco PR; Marco C; González-Pacanowska D; Ruiz-Pérez LM; Carrasco-Jiménez MP; López-Cara LC, 1,2-Diphenoxiethane salts as potent antiplasmodial agents, Bioorganic and Medicinal Chemistry Letters, 2018, Vol. 28: 2485-2489, ARTÍCULO, Id:722898

-Frederick Annang; Ignacio Pérez-Victoria; Theresa Appiah; Guiomar Pérez-Moreno; Elizabeth Domingo; Jesús Martín; Thomas Mackenzie; Luis Ruiz-Pérez; Dolores González-Pacanowska; Olga Genilloud; Francisca Vicente; Christian Agyare; Fernando Reyes, Antiprotozoan sesterterpenes and triterpenes isolated from two Ghanaian mushrooms, Fitoterapia, 2018, Vol. 127: 341-348, ARTÍCULO, Id:717883

DOCTORAL THESES LAST 5 YEARS