US-12622955-B2 - Consensus sequence of the antigen telomerase and the use thereof in preventive and therapeutic vaccination

Abstract

A consensus sequence of the antigen telomerase (ConTRt) can be generated and used in preventive and therapeutic vaccination. The consensus sequence of telomerase was generated by the fusion of two sequences, one belonging to human telomerase (hTERT) and the other to dog telomerase (dTERT), with the aim of developing an effective vaccine for the treatment of tumors expressing both human and dog telomerase, hence in both the human and veterinary sectors.

Inventors

• Luigi Aurisicchio
• Antonella Conforti

Assignees

• EVVIVAX S.R.L.

Dates

Publication Date

20260512

Application Date

20210726

Priority Date

20200729

Claims (11)

1. 1 . A nucleotide sequence encoding an amino acid consensus sequence of the antigen telomerase comprising sequence SEQ ID NO:1, wherein the nucleotide sequence comprises the nucleotide sequence SEQ ID NO: 4, or a sequence having a sequence identity of at least 80% with respect to SEQ ID NO:4, and a nucleotide sequence encoding the profilin-like protein of Toxoplasma Gondii , wherein the nucleotide sequence encoding the profilin-like protein of Toxoplasma Gondii comprises SEQ ID NO: 31.
2. 2 . An expression vector comprising the nucleotide sequence as defined in claim 1 .
3. 3 . The expression vector according to claim 2 , wherein said vector is selected from the group consisting of a plasmid, an RNA, a replicating RNA, amplicons obtained by PCR, and a viral vector.
4. 4 . A DNA, or RNA based vaccine comprising the nucleotide sequence of claim 1 .
5. 5 . A method of vaccinating a subject against telomerase-expressing tumors or treating a subject having a telomerase-expressing tumors, comprising administering the vaccine of claim 4 to the subject.
6. 6 . The method according to claim 5 , wherein the vaccine is administered to the subject by electroporation.
7. 7 . A pharmaceutical composition comprising a nucleotide sequence as defined in claim 1 , in combination with one or more excipients and/or adjuvants.
8. 8 . The nucleotide sequence according to claim 1 , wherein the amino acid sequence further comprises one or more leader sequences.
9. 9 . The nucleotide sequence according to claim 8 , wherein the one or more leader sequence is a secretion leader sequence of a protein selected from the group consisting of tissue plasminogen activator (TPA), IgK, growth hormone, serum albumin, and alkaline phosphatase.
10. 10 . The nucleotide sequence according to claim 1 , wherein the amino acid sequence further comprises one or more immunomodulating amino acid sequences.
11. 11 . The nucleotide sequence according to claim 10 , wherein the one or more immunomodulating amino acid sequences are selected from the group consisting of the fragment crystallisable (Fc) region, profilin-like protein of Toxoplasma Gondii (PFTG) or a functional fragment derived therefrom, the B subunit of the heat-labile toxin of Escherichia Coli (LTB) and the tetanus toxin (TT).

Description

PRIORITY AND CROSS REFERENCE TO RELATED APPLICATIONS This application is the U.S. National Phase Application under 35 U.S.C. § 371 of International Application No. PCT/IT2021/050227, filed Jul. 26, 2021, and published as WO2022/024156A2 on Feb. 3, 2022, which claims the benefit of Foreign Application No. IT102020000018379, filed Jul. 29, 2020. Any and all applications for which a foreign or a domestic priority is claimed is/are identified in the Application Data Sheet filed herewith and is/are hereby incorporated by reference in their entireties under 37 C.F.R. § 1.57. REFERENCE TO SEQUENCE LISTING The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled SequenceListing BARZ070.001APC.txt, created Jan. 26, 2023, which is approximately 84 kilobytes in size, which is replaced by a Replacement Electronic Sequence Listing submitted as herewith a file entitled BARZ070.001ACPCorrectedSequenceListing.txt, which is 76,442 bytes in size and was created on Jul. 17, 2024. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety. FIELD The present invention relates to a consensus sequence of the antigen telomerase and the use thereof in preventive and therapeutic vaccination. In particular, the present invention relates to the generation of a consensus sequence of the antigen telomerase (ConTRt) and the use thereof in preventive and therapeutic vaccination, wherein the telomerase consensus sequence was generated by the fusion of two sequences, one belonging to human telomerase (hTERT) and the other to dog telomerase (dTERT), with the aim of developing an effective vaccine for the treatment of tumours expressing both human and dog telomerase, hence in both the human and veterinary sectors. BACKGROUND It is well known that vaccination is a powerful tool for obtaining the activation of the immune system against pathogens of every type. Vaccination represents the first-line preventive intervention for eliminating the risk of contracting dangerous infectious diseases which can spread throughout a large part of the population, at times causing true epidemics. At present, thanks to vaccines it is possible to defeat highly dangerous infectious diseases for which a therapy either does not exist (poliomyelitis and hepatitis B) or is not always effective (diphtheria, tetanus) or diseases that can be a cause of serious complications (measles, German measles and whooping cough). Immunotherapy, i.e. the therapeutic treatment of disease by means of an activation or suppression of the immune system, has demonstrated to be effective also against various types of tumours, despite having to overcome several limitations, such as immunological tolerance and low immunogenicity. The objective of tumour immunotherapy is therefore to restore the ability of the immune system to recognise tumour cells and eliminate them effectively, by overcoming the mechanisms whereby tumours suppress the immune response. To this end, the immune system recognises tumour-associated molecules (tumour-associated antigens) [Liu 2017], which, despite also being expressed by normal cells, appear on tumour cells in an abnormal manner in terms of quantity, site or time (for example, the carcinoembryonic antigen CEA) or molecules expressed exclusively by tumour cells (tumour-specific antigens), such as the products of normally silent genes [Seremet 2011]. Notwithstanding the discovery of tumour antigens as a target of immunotherapeutic strategies, the greatest obstacle in the development of cancer immunotherapy has been the absence of an antigen common to the majority of patients affected by the most common types of cancer, and thus different research groups have concentrated on the search for a universal tumour-associated antigen capable of inducing a T-cell response of a cytotoxic type (CTL) against a wide range of tumour types. The tumour-associated antigen telomerase represents an ideal candidate for anti-tumour immunotherapy. Telomerase is an enzyme responsible for the lengthening of DNA telomeres, a phenomenon typical of actively proliferating cells, such as embryonal ones, and which is by contrast absent under physiological conditions in somatic cells. Under certain pathological conditions, in which proliferative activity increases, such as in neoplasms, telomerase activity also increases enormously, and various studies have by now demonstrated the correlation existing between telomerase activity and cancer [Akincilar 2016]. In addition to being expressed in about 90% of tumour types, telomerase is degraded by cellular proteasomes into peptides then presented in the context of MHC class I on the surface of the tumour cell as the target of the activity of antigen-specific cytotoxic T-cells. Considering the considerably lower levels of telomerase activity in somatic cells under physiological conditions, the number of spec