De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication
- 1University of California San Diego
Abstract
Malaria kills {approx}600,000 people annually despite recently deployed vaccines. The parasite's blood-stage invasion of erythrocytes, driven by the essential, non-redundant interaction between Plasmodium falciparum RH5 and erythrocyte basigin, remains a leading therapeutic target. Here, we designed de novo mini-proteins (minibinders) to occlude this interface. From {approx}1,500 computationally generated designs, we synthesized 18 candidates ranging from 9.3 to 16.9 kDa which were easily expressed in E. coli as soluble, well-folded proteins. Of these, 7 minibinders blocked blood-stage replication of P. falciparum asexual stage parasites with greater potency than {approx}300nM IC50. Three potent candidate minibinders, mb-5, mb-7, and mb-21, were confirmed to bind RH5 via SPR and BLI and block the merozoite-to-ring transition in stage-synchronized parasite progression assays, with CryoEM of minibinder-RH5 complex confirming the predicted binding at RH5-basigin interface. Although pharmacological challenges remain, our data demonstrate that potent inhibitors of parasite growth, rivaling existing antimalarials, can be designed rather than discovered via extensive screening. Our data highlight the potential for artificial intelligence to change the development landscape of new antimalarial therapies.
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BibTeX
@misc{hsiao2026novo,
title = {{De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication}},
author = {J. Hsiao and M. Shamshoum and A. Chalamalasetty and J. R. Barrett and R. Almada-Monter and E. Tabornal and E. Wilks and Y.-Z. Lee and N. Beutler and A. R. Lukens and P. Kapoor and D. Chen and J. Ndihokubwayo and B. G. Williams and K. McHugh and D. Pulido and I. M. de Sousa and E. Komives and E. V. Noormahomed and I. A. Wilson and D. F. Wirth and T. Rogers and N. Bethel and S. J. Draper and A. Ward and E. A. Winzeler and A. Jinich},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.28.755224},
url = {https://www.biorxiv.org/content/10.64898/2026.09.28.755224v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - De novo minibinders targeting RH5 achieve nanomolar inhibition of blood-stage malaria replication
AU - J. Hsiao
AU - M. Shamshoum
AU - A. Chalamalasetty
AU - J. R. Barrett
AU - R. Almada-Monter
AU - E. Tabornal
AU - E. Wilks
AU - Y.-Z. Lee
AU - N. Beutler
AU - A. R. Lukens
AU - P. Kapoor
AU - D. Chen
AU - J. Ndihokubwayo
AU - B. G. Williams
AU - K. McHugh
AU - D. Pulido
AU - I. M. de Sousa
AU - E. Komives
AU - E. V. Noormahomed
AU - I. A. Wilson
AU - D. F. Wirth
AU - T. Rogers
AU - N. Bethel
AU - S. J. Draper
AU - A. Ward
AU - E. A. Winzeler
AU - A. Jinich
PY - 2026
DA - 2026/09/28
PB - bioRxiv
DO - 10.64898/2026.09.28.755224
UR - https://www.biorxiv.org/content/10.64898/2026.09.28.755224v1
AB - Malaria kills {approx}600,000 people annually despite recently deployed vaccines. The parasite's blood-stage invasion of erythrocytes, driven by the essential, non-redundant interaction between Plasmodium falciparum RH5 and erythrocyte basigin, remains a leading therapeutic target. Here, we designed de novo mini-proteins (minibinders) to occlude this interface. From {approx}1,500 computationally generated designs, we synthesized 18 candidates ranging from 9.3 to 16.9 kDa which were easily expressed in E. coli as soluble, well-folded proteins. Of these, 7 minibinders blocked blood-stage replication of P. falciparum asexual stage parasites with greater potency than {approx}300nM IC50. Three potent candidate minibinders, mb-5, mb-7, and mb-21, were confirmed to bind RH5 via SPR and BLI and block the merozoite-to-ring transition in stage-synchronized parasite progression assays, with CryoEM of minibinder-RH5 complex confirming the predicted binding at RH5-basigin interface. Although pharmacological challenges remain, our data demonstrate that potent inhibitors of parasite growth, rivaling existing antimalarials, can be designed rather than discovered via extensive screening. Our data highlight the potential for artificial intelligence to change the development landscape of new antimalarial therapies.
ER -