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BR-102024017417-A2 - Association of recombinant proteins RASP-2 and RTC24 as a vaccine strategy against Chagas disease.

BR102024017417A2BR 102024017417 A2BR102024017417 A2BR 102024017417A2BR-102024017417-A2

Abstract

Chagas disease represents a global public health problem, with socioeconomic losses resulting from long-term treatment due to persistent infection by the protozoan Trypanosoma cruzi. In this context, the search for efficient prophylactic measures to control this disease becomes necessary. The use of recombinant proteins in vaccine formulations has been widely studied, presenting itself as an efficient and safe alternative for the control of various infectious diseases. Thus, the present invention refers to the combined use of the recombinant proteins rTC-24 and rASP-2 of Trypanosoma cruzi in vaccine formulations, using them as a control strategy for Chagas disease. The invention comprises the use of the aforementioned proteins, in combination, expressed in Escherichia coli, for use in vaccine formulations. The invention described herein meets the criteria described for the technical sectors: A61K 39/00; A61K 39/002 and A61K 93/005.

Inventors

  • Sibele Borsuk
  • FERNANDA SEVERO SABEDRA SOUZA
  • BÁRBARA DA ROCHA FONSECA
  • GUILHERME SENNA DOS SANTOS
  • LUIZA DOMINGUES MORON
  • MIRNA SAMARA DIÉ ALVES
  • FERNANDA KANAAN DE AZAMBUJA
  • GUSTAVO DOS SANTOS HARTLEBEN
  • Fabiana Kommling Seixas
  • TIAGO COLLARES

Assignees

  • UNIVERSIDADE FEDERAL DE PELOTAS

Dates

Publication Date
20260310
Application Date
20240823

Claims (4)

  1. 1. Vaccine formulation against Trypanosoma cruzi characterized by the use of recombinant proteins: amastigote surface protein no. 2, rASP-2, (SEQ ID NO:1) and 24 kDa flagellar calcium-binding protein, rTC24, (SEQ ID NO:2) expressed in Escherichia coli.
  2. 2. Vaccine formulation against Trypanosoma cruzi, according to claim 1, characterized by the use of recombinant proteins rASP-2 and rTC24 in combination and in isolation, together with commercial adjuvants.
  3. 3. Vaccine formulation against Trypanosoma cruzi, according to claim 1, characterized by the use of recombinant proteins rASP-2 and rTC24 in combination and in isolation, for use in in vivo tests in different guinea pig models.
  4. 4. Vaccine formulation against Trypanosoma cruzi, according to claim 2, characterized by the use of recombinant proteins rASP-2 and rTC24 in combination and in isolation, for use as vaccine formulations for human use against Chagas disease.

Description

Field of invention [001] The present invention consists of the use of recombinant Trypanosoma cruzi antigens in vaccine formulations as a prophylactic strategy for Chagas disease. The proteins in question consist of the terminal portion (from amino acid 261) of amastigote surface protein no. 2 (ASP-2) and the 24 kDa calcium-binding flagellar protein (TC24), in their complete form. [002] These proteins will be recombinantly produced and expressed from Escherichia coli bacteria using the pAE vector. The nucleotide sequences for these proteins are available in the NCBI database www.ncbi.nlm.nih.gov (AAC47720.1; KJ668043.1; AAB08762.1). From this, the sequences SEQ ID NO:1 and SEQ ID NO:2 were obtained for the rASP-2 and rTC24 proteins, respectively. [003] Fundamentals of the invention [004] Chagas disease (CD), also known as American trypanosomiasis, is a parasitic infection caused by the flagellated protozoan Trypanosoma cruzi. The main form of transmission occurs through Triatomine insects, which act as biological vectors of this disease. Infection occurs through contact with the contaminated feces of the insect, reaching the bloodstream, allowing the parasites to multiply and establish the infection. Furthermore, other forms of transmission include congenital transmission, blood transfusion, organ transplants, or ingestion of contaminated food or beverages (J. R. Coura, The main scenarios of Chagas disease transmission. The vectors, blood and oral transmissions - A comprehensive review, Mem Inst Oswaldo Cruz, vol. 110, no. 3, p. 277-282, 2015, doi: 10.1590/0074-0276140362; A. Rassi, A. Rassi, and J. A. Marin-Neto, Chagas disease, The Lancet, vol. 375, no. 9723, Elsevier B.V., p. 1388-1402, 2010. doi: 10.1016/S0140-6736(10)60061-X.). [005] T. cruzi has a complex life cycle, exhibiting three distinct forms and consisting of an invertebrate host (insect) and a vertebrate host, with humans being the most relevant species. The trypomastigote stage represents the infective phase, present in triatomine bugs and also the circulating phase in the definitive host organism. Upon infecting new cells, the parasite internalizes its flagellum, assumes the amastigote form, and acquires reproductive capacity, allowing its multiplication by binary fission. Differentiation to the trypomastigote form occurs again when released into the bloodstream to infect new cells, also characterizing the diagnostic phase. Insect infection occurs through the ingestion of blood contaminated with circulating trypomastigote parasites, which transform into epimastigotes in the vector's digestive tract, where they also exhibit reproductive capacity, multiplying and again assuming the infective metacyclic trypomastigote form (CDC, Centers for Disease Control and Prevention, June 2021. Available at: https://www.cdc.gov/dpdx/trypanosomiasisamerican/index.html. Accessed on: June 5, 2024). [006] According to epidemiological data released by the World Health Organization in 2018, the incidence is estimated at 6 to 8 million cases, with the highest concentration in Latin America (World Health Organization, 2018). The main factor linked to the concentration of cases in this region is the low investment in research and adequate prevention measures, with a higher incidence in rural areas, where access to diagnosis and treatment is difficult. For this reason, it is characterized by the WHO as a neglected tropical disease (B. de P. Fonseca, P. C. Albuquerque, and F. Zicker, Neglected tropical diseases in Brazil: lack of correlation between disease burden, research funding and output, Tropical Medicine and International Health, vol. 25, no. 11, pp. 1373–1384, Nov. 2020, doi: 10.1111/tmi.13478; F. R. Martins-Melo, M. Carneiro, A. N. Ramos, J. Heukelbach, A. L. P. Ribeiro, and G. L. Werneck, The burden of Neglected Tropical Diseases in Brazil, 1990–2016: A subnational analysis from the Global Burden of Disease Study 2016, PLoS Negl Trop Dis, vol. 12, no. 6, Jun. 2018, doi: 10.1371/journal.pntd.0006559.). [007] After infection is established, Chagas disease develops in different phases. The acute phase represents the initial stage of the disease, which may or may not be symptomatic, lasting for weeks or months. With the immune system acting, parasite levels in the body remain stable, without evident clinical signs, but still diagnosable. This period is known as the indeterminate phase, and these conditions can persist until the end of life. The chronic phase is represented, in some cases, by the development of abnormalities in organs of the gastrointestinal tract (megacolon or megaesophagus) or damage to the cardiovascular system (arrhythmias, thromboembolism) and heart organs (cardiomegaly). However, it can remain asymptomatic or without pathological organ changes until the end of the host's life (J. Guarner, Chagas disease as an example of a reemerging parasite, Seminars in Diagnostic Pathology, vol. 36, no. 3. W.B. Saunders, pp. 164-169, May 1, 2019. doi: 10