[go: up one dir, main page]

WO2017163171A1 - Protéine chimère, composition vaccinale contre les leishmanioses et utilisations - Google Patents

Protéine chimère, composition vaccinale contre les leishmanioses et utilisations Download PDF

Info

Publication number
WO2017163171A1
WO2017163171A1 PCT/IB2017/051610 IB2017051610W WO2017163171A1 WO 2017163171 A1 WO2017163171 A1 WO 2017163171A1 IB 2017051610 W IB2017051610 W IB 2017051610W WO 2017163171 A1 WO2017163171 A1 WO 2017163171A1
Authority
WO
WIPO (PCT)
Prior art keywords
saponin
chimera
leishmaniasis
protein
sla
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/IB2017/051610
Other languages
English (en)
Portuguese (pt)
Inventor
Eduardo ANTONIO FERRAZ COELHO
Carlos ALBERTO PEREIRA TAVARES
Vivian TAMIETTI MARTINS
Tiago Antônio De Oliveira MENDES
Mariana COSTA DUARTE
Daniel Menezes SOUZA
Bruno MENDES ROATT
Daniela PAGLIARA LAGE
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Universidade Federal de Minas Gerais
Original Assignee
Universidade Federal de Minas Gerais
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Universidade Federal de Minas Gerais filed Critical Universidade Federal de Minas Gerais
Publication of WO2017163171A1 publication Critical patent/WO2017163171A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/002—Protozoa antigens
    • A61K39/008—Leishmania antigens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/44—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from protozoa
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to a vaccine composition based on a recombinant polypeptide chimera composed of human and mouse CD4 + and CD8 + T-lymphocyte specific epitopes derived from four proteins (LiHypl, LiHyp6, LiHyV and HRF) from Leishmania, which was able to induce protection against visceral and cutaneous leishmaniasis and its use.
  • a recombinant polypeptide chimera composed of human and mouse CD4 + and CD8 + T-lymphocyte specific epitopes derived from four proteins (LiHypl, LiHyp6, LiHyV and HRF) from Leishmania, which was able to induce protection against visceral and cutaneous leishmaniasis and its use.
  • Leishmaniasis are endemic infectious diseases in various tropical and subtropical regions on different continents. It is estimated that 380 million people are at risk of contracting the disease and there is an approximate incidence of 0.5 to 1, 0 and 1, 5 to 2.0 million new cases of visceral leishmaniasis per year ( VL) and cutaneous leishmaniasis (LT), respectively, worldwide. Brazil is the leading VL incidence site in the Americas, accounting for 95% of cases (Alvar J, et al. Leishmaniasis worldwide and global estimates of its incidence. PLoS One. 7: e35671, 2012).
  • an effective vaccine should contain immunogenic antigens conserved on different Leishmania species, and have an affordable cost to the population and their governments.
  • immunogenic antigens conserved on different Leishmania species, and have an affordable cost to the population and their governments.
  • most studies performed use unique antigens, which are expressed in only one of the morphological forms of the parasite (Fernandes AP, et al. Making an anti-amastigote vaccine for visceral leishmaniasis: rational, update and perspectives. Curr Opin Microbiol. 15 : 1-10, 2012).
  • LiHypl XP_001468941 .1
  • LiHyp6 XP_001568689.1
  • HRF CAJ05086.1
  • LiHyV protein (XP_001462854.1) offered recombinant and saponin-associated protection against infection with this same parasite species (Martins VT, et al. Leishmania-specific hypothetical protein expressed in both promastigote and amastigote stages of Leishmania infantum employed for the serodiagnosis of, and as a vaccine candidate against, visceral leishmaniasis (Parasit. Vectors, 8; 363, 2015); thus demonstrating the feasibility of using such antigens as candidate vaccines against LV.
  • Patent document BR132013001 271 entitled Chimeric Protein, Vaccine Composition and Visceral Leishmaniasis Immunodiagnostic Test Kit, refers to a chimeric protein for use in canine and human LV immunodiagnostic vaccine and test composition developed through selection, identification , production and testing of new antigens by analysis proteomics, bioinformatics, peptide synthesis, immunoassay, synthetic gene construction and protein production and purification.
  • Such technology differs from the present invention in that it comprises other immunogenic components which are different from those presented herein.
  • the patent document P 10,609,847, entitled ⁇ Composition Comprising the N-terminal region of Leishmania histone H2B, use thereof for inducing an immune response _ is an immunogenic composition comprising an antigenic peptide consisting of a fragment of histone protein H2B Leishmania is an adjuvant that stimulates the immune response.
  • This technology differs from the present invention in that it comprises only one antigenic compound.
  • the antigen in question differs from any of those described in the present invention.
  • Patent document WO20131 10824 entitled Multi-component ⁇ Chimera for use as a vaccine against infection by Leishmania spp.
  • mammals refers to a chimera, HISA70, composed of different proteins (H2A, H2B, H3, H4, A2 and HSP70) from Leishmania infantum in the composition of a vaccine against Leishmania spp.
  • HISA70 composed of different proteins (H2A, H2B, H3, H4, A2 and HSP70) from Leishmania infantum in the composition of a vaccine against Leishmania spp.
  • Such technology differs from the present invention in that it comprises proteins other than those listed in the present invention.
  • the present invention relates to a chimeric polypeptide vaccine composed of human and mouse CD4 + and CD8 + T-lymphocyte specific epitopes derived from the four proteins (LiHypl, LiHyp6, LiHyV and HRF) that have been described as capable. to induce protection, when administered alone, against L. infantum infection, to induce protection against infection by L. infantum and L. amazonensis, species causing visceral and cutaneous leishmaniasis, respectively, worldwide.
  • the polypeptide chimera has shown promising results in inducing protection against challenge infection with both parasite species and, thus, is a new candidate for protection against distinct Leishmania species.
  • the protection afforded by the chimeric vaccine was superior to that induced by proteins administered alone in the experimental animals with respect to the immunogenicity induced before and after experimental infection, as well as reducing the parasitic load in different organs of the evaluated animals.
  • Figure 1 represents the antigenicity of recombinant chimera.
  • the recombinant chimera (10 ⁇ g, indicated by the black arrow in the figure) was electrophoresed on a 12% SDS-PAGE denaturing gel, stained with bright Coomassie blue G-250 and transferred to a nitrocellulose membrane, which was placed. incubation with the different serum pools. For this, a molecular weight standard was used as marker (A).
  • Reactions with the serum pools of animals immunized with recombinant proteins LiHypl (B), LiHyp6 (C), LiHyV (D), HRF (E) or sera from animals immunized with the chimera itself (F) are shown.
  • Serum pools of mice experimentally infected with L. infantum (G) or L. amazonensis (H) were also used, as well as sera pool from uninfected and unimmunized animals (I), used as negative reaction control.
  • the present invention relates to a polypeptide chimera-containing vaccine composition composed of human and mouse CD4 + and CD8 + T lymphocyte specific epitopes from four Leishmania immunogenic proteins and their use for the protection and / or treatment against leishmaniasis cutaneous and visceral.
  • the leishmaniasis vaccine composition comprises the chimeric protein defined by SEQ ID NO: 1, in addition to the association of pharmaceutically acceptable adjuvants.
  • adjuvants may be selected, without limitation, from those cited in the following literature: Remington's Pharmaceutical Sciences, Mack Publishing, European or Brazilian Pharmacopoeia or new excipients to be preferably saponins.
  • adjuvants should be Th1 immune response inducers, such as saponins, or compounds that are capable of stimulating a cellular immune response prime by the production of IFN- ⁇ and IL-12.
  • the present invention further relates to the use of the chimeric protein in tegumentary and visceral leishmaniasis vaccine compositions.
  • the proposed vaccine composition may be administered by the intradermal, intramuscular, oral, nasal, intravenous, subcutaneous and / or as a device that can be implanted and / or injected.
  • the proteins LiHypl, LiHyp6, LiHyV and HRF were subjected to bioinformatics analysis for selection of immunogenic regions rich in murine and human T lymphocyte epitopes.
  • the Net CTL Pan program was used, so that the best epitopes that bound to the human MHC class I alleles A2, A3 and B7, which together represent over 90% of the human population of any ethnicity, were selected.
  • the same program was used so that epitopes binding to the ⁇ -2-Kd, ⁇ -2-Ld and ⁇ -2-Dd alleles of BALB / c mice were also selected.
  • the epitopes specific to the class I major histocompatibility complex (MHC) molecules that were selected for chimera construction are shown in Tables 1 and 2.
  • the Net MHC II 2.2 program was used to assess the binding affinity of 15 amino acid peptides to the 26 human alleles. Only epitopes with the ability to interact with binding affinity of less than 500 nM and in at least 30% of alleles were selected. The program was also used to select epitopes that bound to the 1-Ad and 1-Ed alleles of BALB / c mice. The epitopes specific to MHC class II molecules that have been selected for chimera construction are shown in Tables 3 and 4. Table 1. In silico prediction of specific epitopes to MHC class I T-lymphocyte molecules from humans selected for recombinant chimera construction.
  • the immunogenicity of recombinant chimera was evaluated in immunized BALB / c mice by assaying IFN- ⁇ , IL-12, GM-CSF, IL-4 and IL-10 cytokines produced by animal splenocytes, 30 days after administration of the last dose of the immunogen.
  • IFN- ⁇ , IL-12, GM-CSF, IL-4 and IL-10 cytokines produced by animal splenocytes 30 days after administration of the last dose of the immunogen.
  • SLA serum-associated cytokines
  • the chimera plus saponin immunized group produced higher levels of IFN- ⁇ , IL-12 and GM-CSF than the levels of these cytokines obtained in the groups immunized with rLiHypl, rLiHyp6, rLiHyV or rHRF plus saponin proteins. No increase was observed in IL-4 and IL-10 production in any of the groups evaluated.
  • the chimera plus saponin immunized group produced higher levels of IFN- ⁇ , IL-12 and GM-CSF than the levels of these cytokines obtained in the groups immunized with rLiHypl, rLiHyp6, rLiHyV or rHRF proteins. saponin.
  • animals in the control groups (saline and saponin) produced significantly higher levels of IL-4 and IL-10 than the other groups evaluated (Table 9).
  • Table 8 Levels of production of IFN- ⁇ , IL-12 and GM-CSF cytokines (pg / ml). Cytokines were quantified in animal splenocyte culture supernatants. The mean ⁇ standard deviation of the groups is shown.
  • Table 9- IL-4 and IL-10 cytokine production levels (in pg / ml). Cytokines were quantified in animal splenocyte culture supernatants. The mean ⁇ standard deviation of the groups is shown.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Veterinary Medicine (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

La présente invention concerne une composition vaccinale à base d'une chimère polypeptidique recombinante constituée par desépitopes spécifiques de lymphocytes T CD4+ et CD8+ d'être humain et de souris dérivés de quatre protéines de (LiHypl, LiHyp6, LiHyV e HRF) de Leishmania, capable d'induire une protection contre la leishmaniose viscérale et tégumentaire, ainsi que son utilisation.
PCT/IB2017/051610 2016-03-21 2017-03-20 Protéine chimère, composition vaccinale contre les leishmanioses et utilisations Ceased WO2017163171A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
BR102016006121-0A BR102016006121A2 (pt) 2016-03-21 2016-03-21 Chemical protein, vaccine composition against leishmaniosis and uses
BRBR1020160061210 2016-03-21

Publications (1)

Publication Number Publication Date
WO2017163171A1 true WO2017163171A1 (fr) 2017-09-28

Family

ID=59914225

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IB2017/051610 Ceased WO2017163171A1 (fr) 2016-03-21 2017-03-20 Protéine chimère, composition vaccinale contre les leishmanioses et utilisations

Country Status (2)

Country Link
BR (1) BR102016006121A2 (fr)
WO (1) WO2017163171A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019159196A1 (fr) * 2018-02-15 2019-08-22 National Centre For Cell Science Nouvelle protéine chimérique kinase-c en tant qu'immunomodulateur

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025118050A1 (fr) * 2023-12-04 2025-06-12 Laboratório Bio-Vet Ltda Antigène vaccinal chimérique à action triple contre l'ehrlichiose, l'anaplasmose et la babésiose du chien, vecteur d'expression, cellule recombinante, composition vaccinale, kit de diagnostic, méthode de traitement et utilisation d'une protéine chimérique

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
MARTINS, V. T. ET AL.: "A Leishmania-specific hypothetical protein expressed in both promastigote and amastigote stages of Leishmania infantum employed for the serodiagnosis of, and as a vaccine candidate against, visceral leishmaniasis", PARASIT VECTORS., vol. 8, no. 363, July 2015 (2015-07-01) *
MARTINS, V. T. ET AL.: "Antigenicity and Protective Efficacy of a Leishmania Amastigote-specific Protein, Member of the Super- oxygenase Family, against Visceral Leishmaniasis", PLOS NEGL TROP DIS., vol. 7, no. 3, 2013, pages e2148 *
MARTINS, V. T. ET AL.: "Antigenicity, Immunogenicity and Protective Efficacy of Three Proteins Expressed in the Promastigote and Amastigote Stages of Leishmania infantum against Visceral Leishmaniasis", PLOS ONE, vol. 10, no. 9, 14 September 2016 (2016-09-14) *
MARTINS, V., T. ET AL.: "A recombinant chimeric protein composed by human and mice-specific CD 4 and CD 8 T cell epitopes protects against visceral leishmaniasis", PARASITE IMMUNOLOGY, vol. 12359, 16 September 2016 (2016-09-16), pages 1 - 14 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019159196A1 (fr) * 2018-02-15 2019-08-22 National Centre For Cell Science Nouvelle protéine chimérique kinase-c en tant qu'immunomodulateur

Also Published As

Publication number Publication date
BR102016006121A2 (pt) 2017-09-26

Similar Documents

Publication Publication Date Title
ES2670859T3 (es) Secuencias peptídicas y composiciones de Anopheles gambiae
Solioz et al. The protective capacities of histone H1 against experimental murine cutaneous leishmaniasis
Shokri et al. Comparing Montanide ISA 720 and 50-V2 adjuvants formulated with LmSTI1 protein of Leishmania major indicated the potential cytokine patterns for induction of protective immune responses in BALB/c mice
ES2760004T3 (es) Vacuna contra Leishmania utilizando un inmunnógeno salival de mosca de la arena
BRPI0719078A2 (pt) Vacina contra leishmania canina
WO2017163171A1 (fr) Protéine chimère, composition vaccinale contre les leishmanioses et utilisations
Ribeiro et al. Evaluation of a Leishmania hypothetical protein administered as DNA vaccine or recombinant protein against Leishmania infantum infection and its immunogenicity in humans
BRPI1015136B1 (pt) Vacina contra esquistossomose
BR112015015539A2 (pt) Vacinas monovalentes e polivalentes contra leishmania
BR102020021166A2 (pt) Composições farmacêuticas e/ou imunogênicas contra leishmaniose visceral e uso
BR102017025621A2 (pt) Proteína quimérica recombinante, vacina contra leishmanioses e uso
BR102020008111A2 (pt) Composição vacinal contra leishmaniose visceral e uso
BR102018070807A2 (pt) Proteína quimérica recombinante, vacina contra leishmaniose tegumentar e visceral e usos
BR102018016046A2 (pt) Composições vacinais contra as leishmanioses tegumentar e visceral e uso
WO2018145182A1 (fr) Peptide hybride, ensemble de peptides hybrides, composition, utilisations du peptide hybride, méthode d'induction d'une réponse immunitaire et trousses
Al-Idreesi et al. Immune responses to Eimeria tenella sporozoite protein as vaccine to broiler against coccidiosis
WO2023077206A1 (fr) Épitopes et protéines multiépitopes génétiquement modifiés, compositions immunogènes et utilisation de celles-ci
BR102014022027B1 (pt) Imunobiológico para controle do vetor da leishmaniose, processos de obtenção e usos
BR102021010469A2 (pt) Proteína quimérica recombinante, seu uso, e, composição
BR102022022571A2 (pt) Epítopos e proteínas multiepítopos geneticamente modificados, composições imunogênicas e uso das mesmas
BR112016012097A2 (pt) composição vacinal para a prevenção e/ou o tratamento de leishmanioses, peptídeos imunogênicos e processo de obtenção
AU2013200258B2 (en) Peptide sequences and compositions
WO2012061910A2 (fr) Domaines et épitopes immunoprotecteurs principaux de la nucléoside hydrolase de leishmania (l) donovani, à utiliser dans la vaccination, l'immunothérapie et le diagnostic
BRPI1015788B1 (pt) principais domínios e epítopos imunoprotetores da nucleosídeo hidrolase de leishmania (l.) donovani para uso em vacinação
JP2002533474A (ja) エル．インファンタム（Ｌ．ｉｎｆａｎｔｕｍ）の４つのタンパク質の抗原決定基をコードするキメラ遺伝子

Legal Events

Date Code Title Description
NENP Non-entry into the national phase

Ref country code: DE

121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17769543

Country of ref document: EP

Kind code of ref document: A1

122 Ep: pct application non-entry in european phase

Ref document number: 17769543

Country of ref document: EP

Kind code of ref document: A1