Hybrid peptide, set of hybrid peptides, composition, uses of the hybrid peptide, method for inducing an immune response and kits
Inventors
Chudzinski-Tavassi, Ana Marisa • Vaz Galväo da Silva, Bárbara Athayde • Mesquita Pasqualoto, Kerly Fernanda • IWAI, LEO KEI • Porto, Rafael Marques
Assignees
Imunotick Pesquisa E Desenvolvimento Tecnologico SA • Fundacao Butantan
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Publication Number
US-11034735-B2
Publication Date
2021-06-15
Expiration Date
Abstract
The present invention relates to novel hybrid peptides developed from the combination of membrane surface protein peptide fragments present in anaplasmosis microorganisms, and in particular, bacteria of the species Anaplasma marginale. The present invention further relates to hybrid peptide sets, compositions and kits comprising such novel hybrid peptides, their uses and methods of inducing immune response. Each hybrid peptide, according to the present invention, comprises two or more peptide fragments of amino acid sequences as defined in the present invention linked together by means of a spacer element. The combined peptide fragments are protein peptide fragments of MSP1, MSP1a, MSP1b, MSP2, MSP2-HRV, MSP3, OMP7, OMP8, VirB9 and VirB10.
Core Innovation
The invention relates to a vaccine focused on Anaplasma marginale using synthetic hybrid peptides. Hybrid peptides are formed by linking defined peptide fragments selected from MSP and OMP peptide fragments and Type IV secretion system factor fragments VirB9 and VirB10, using spacer elements. The spacer elements connect the fragments into a single peptide.
The hybrid peptide constructs include at least one hybrid peptide comprising two peptide fragments consisting of amino acid sequences defined by SEQ ID NOs and joined by a spacer element. Hybrid peptide sets are provided that include at least two hybrid peptides, where the fragment-pairing differs between the hybrid peptides. Spacer elements can be constrained to particular compositions, including preferred spacer elements containing glycine and proline residues.
Compositions and kits are disclosed that include the hybrid peptide sets and optionally one or more adjuvants, with formulation forms such as solution, mixture, powder, aerosol, or lyophilized compositions. The invention further discloses methods for inducing an immune response by administering therapeutically effective amounts of the hybrid peptides to a human or animal, and documented immune-response induction and evaluation include immunogenicity testing using assays such as Dot-ELISA, Western blotting, and indirect immunofluorescence.
Claims Coverage
The claims cover a composition that includes a set of hybrid peptides with specific fragment-linking architectures. The claim set includes one independent claim and multiple dependent refinements defining spacer constraints, additional hybrid peptide compositions, selectable adjuvants, and a dosing or immune-response administration context.
Hybrid peptide composition with defined fragment pairing
A composition comprising at least two hybrid peptides, where at least one hybrid peptide comprises two peptide fragments consisting of amino acid sequences SEQ ID NO: 14 and SEQ ID NO: 15 linked together by a spacer element, and a second hybrid peptide comprises two peptide fragments consisting of amino acid sequences SEQ ID NO: 13 and SEQ ID NO: 19 linked together by a spacer element.
Spacer element with constrained glycine residues
The composition wherein each spacer element consists of 2 to 15 glycine residues.
Additional hybrid peptide with specified fragment identities
The composition including at least one additional hybrid peptide made from two peptide fragments with amino acid sequences SEQ ID NO: 5 and SEQ ID NO: 6 joined by a spacer element.
Number of additional hybrid peptides
The peptide composition wherein the composition includes additional hybrid peptides in an amount ranging from 2 to 8.
Selectable adjuvants
The peptide composition where one or more adjuvants are selected from Freund's complete adjuvant, Freund's incomplete adjuvant, aluminum hydroxide, silica, and saponin.
Therapeutically effective dosing range per hybrid peptide for immune response induction
A method of inducing an immune response by administering a therapeutically effective amount of 50 to 150 μg of each hybrid peptide per human or animal.
The claims require a composition comprising at least two hybrid peptides with defined SEQ ID NO fragment pairings linked by spacer elements. The dependent claim coverage further specifies spacer glycine residue constraints, additional hybrid peptides with specified SEQ ID NO fragment identities, a defined range for the number of additional hybrids, and optional named adjuvants. The disclosed method context specifies therapeutically effective amounts per hybrid peptide for immune response induction in a human or animal.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Immunization, treatment, and protection using the hybrid peptide compositions.
In vitro use.
Diagnosis.
Inducing an immune response by administering therapeutically effective amounts of the hybrid peptides to a human or animal.
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