Nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal for increasing the expression of an encoded pathogenic antigen

Patent metadata

Field	Value
Publication number	US-9669089-B2
Application number	US-201314378538-A
Country	US
Kind code	B2
Filing date	Feb 15, 2013
Priority date	Feb 15, 2012
Publication date	Jun 6, 2017
Grant date	Jun 6, 2017

Abstract

The present invention relates to a nucleic acid sequence, comprising or coding for a coding region, encoding at least one peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, at least one histone stem-loop and a poly(A) sequence or a polyadenylation signal. Furthermore the present invention provides the use of the nucleic acid for increasing the expression of said encoded peptide or protein. It also discloses its use for the preparation of a pharmaceutical composition, especially a vaccine, e.g. for use in the treatment of infectious diseases. The present invention further describes a method for increasing the expression of a peptide or protein comprising a pathogenic antigen or a fragment, variant or derivative thereof, using the nucleic acid comprising or coding for a histone stem-loop and a poly(A) sequence or a polyadenylation signal.

First claim

The invention claimed is:

1. A nucleic acid molecule comprising:
a) a polypeptide coding region, encoding an infectious disease antigen, wherein the infectious disease antigen is not a histone polypeptide;
b) at least one histone stem-loop; and
c) a poly(A) sequence or a polyadenylation signal sequence.

2. The nucleic acid molecule according to claim 1 , wherein the infectious disease antigen is a bacterial, viral, protozoan, or fungal antigen.

3. The nucleic acid molecule of claim 1 , wherein the infectious disease antigen is an antigen from an organism selected from the group consisting of Acinetobacter baumannii, Anaplasma genus, Anaplasma phagocytophilum, Ancylostoma braziliense, Ancylostoma duodenale, Arcanobacterium haemolyticum, Ascaris lumbricoides, Aspergillus genus, Astroviridae, Babesia genus, Bacillus anthracis, Bacillus cereus, Bartonella henselae , BK virus, Blastocystis hominis, Blastomyces dermatitidis, Bordetella pertussis, Borrelia burgdorferi, Borrelia genus, Borrelia spp, Brucella genus, Brugia malayi , Bunyaviridae family, Burkholderia cepacia, Burkholderia mallei, Burkholderia pseudomallei , Caliciviridae family, Campylobacter genus, Candida albicans, Candida spp, Chlamydia trachomatis, Chlamydophila pneumoniae, Chlamydophila psittaci, Clonorchis sinensis, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium perfringens, Clostridium spp, Clostridium tetani, Coccidioides spp, coronaviruses, Corynebacterium diphtheriae, Coxiella burnetii , Crimean-Congo hemorrhagic fever virus, Cryptococcus neoformans, Cryptosporidium genus, Cytomegalovirus (CMV), Dengue viruses, Dientamoeba fragilis , Ebolavirus (EBOV), Echinococcus genus, Ehrlichia chaffeensis, Ehrlichia ewingii, Ehrlichia genus, Entamoeba histolytica, Enterococcus genus, Enterovirus genus, Enteroviruses, Epidermophyton spp, Epstein-Ban Virus (EBV), Escherichia coli 0157:H7, Escherichia coli 0111, Escherichia coli 0104:H4, Fasciola hepatica and Fasciola gigantica , Filarioidea superfamily, Flaviviruses, Francisella tularensis, Fusobacterium genus, Geotrichum candidum, Giardia intestinalis, Gnathostoma spp, GSS prion, Guanarito virus, Haemophilus ducreyi, Haemophilus influenzae, Helicobacter pylori , Henipavirus (Hendra virus Nipah virus), Hepatitis A Virus, Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Hepatitis D Virus, Hepatitis E Virus, Herpes simplex virus 1 (HSV-1), Herpes simplex virus 2 (HSV-2), Histoplasma capsulatum , Human immunodeficiency virus (HIV), Hortaea werneckii , Human bocavirus (HBoV), Human herpesvirus 6 (HHV-6), Human herpesvirus 7 (HHV-7), Human metapneumovirus (hMPV), Human papillomavirus (HPV), Human parainfluenza viruses (HPIV), Japanese encephalitis virus, JC virus, Junin virus, Kingella kingae, Klebsiella granulomatis , Lassa virus, Legionella pneumophila, Leishmania genus, Leptospira genus, Listeria monocytogenes , Lymphocytic choriomeningitis virus (LCMV), Machupo virus, Malassezia spp, Marburg virus, Measles virus, Metagonimus yokagawai , Microsporidia phylum, Molluscum contagiosum virus (MCV), Mumps virus, Mycobacterium leprae and Mycobacterium lepromatosis, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumoniae, Naegleria fowleri, Necator americanus, Neisseria gonorrhoeae, Neisseria meningitidis, Nocardia asteroides, Nocardia spp, Onchocerca volvulus, Orientia tsutsugamushi , Orthomyxoviridae family (Influenza), Paracoccidioides brasiliensis, Paragonimus spp, Paragonimus westermani , Parvovirus B19, Pasteurella genus, Plasmodium genus, Pneumocystis jirovecii , Poliovirus, Rabies virus, Respiratory syncytial virus (RSV), Rhinovirus, rhinoviruses, Rickettsia akari, Rickettsia genus, Rickettsia prowazekii, Rickettsia rickettsii, Rickettsia typhi , Rift Valley fever virus, Rotavirus, Rubella virus, Sabia virus, Salmonella genus, Sarcoptes scabiei , SARS coronavirus, Schistosoma genus, Shigella genus, Sin Nombre virus, Hantavirus, Sporothrix schenckii, Staphylococcus genus, Staphylococcus genus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Strongyloides stercoralis, Taenia genus, Taenia solium , Tick-borne encephalitis virus (TBEV), Toxocara canis, Toxocara cati, Toxoplasma gondii, Treponema pallidum, Trichinella spiralis, Trichomonas vaginalis, Trichophyton spp, Trichuris trichiura, Trypanosoma brucei, Trypanosoma cruzi, Ureaplasma urealyticum , Varicella zoster virus (VZV), Varicella zoster virus (VZV), Variola major or Variola minor, Venezuelan equine encephalitis virus, Vibrio cholerae , West Nile virus, Western equine encephalitis virus, Wuchereria bancrofti , Yellow fever virus, Yersinia enterocolitica, Yersinia pestis , and Yersinia pseudotuberculosis.

4. The nucleic acid molecule of claim 1 , wherein the infectious disease antigen is a prion.

5. The nucleic acid molecule of claim 3 , wherein the infectious disease antigen is an antigen from an organism selected from the group consisting of Influenza virus, respiratory syncytial virus (RSV), Human immunodeficiency virus (HIV), Staphylococcus aureus , Dengue virus, Rabies virus, Rota virus and Yellow Fever Virus.

6. The nucleic acid molecule of claim 5 , wherein the infectious disease antigen is selected from the group consisting of the HIV p24 antigen, HIV envelope protein Gp120, HIV envelope protein Gp41, HIV envelope protein Gp160, HIV polyprotein GAG, HIV negative factor protein Nef, HIV trans-activator of transcription Tat, Dengue virus capsid protein C, Dengue virus premembrane protein prM, Dengue virus membrane protein M, Dengue virus envelope protein E, Dengue virus protein NS1, Dengue virus protein NS2A, Dengue virus protein NS2B, Dengue virus protein NS3, Dengue virus protein NS4A, Dengue virus protein 2K, Dengue virus protein NS4B, Dengue virus protein, Influenza virus Hemagglutinin (HA), Influenza virus Neuraminidase (NA), Influenza virus Nucleoprotein (NP), Influenza virus M1 protein, Influenza virus M2 protein, Influenza virus NS1 protein, Influenza virus NS2 protein, Influenza virus PA protein, Influenza virus PB 1 protein, Influenza virus PB 1-F2 protein, Influenza virus PB2 protein, Rabies virus nucleoprotein N, Rabies virus large structural protein L, Rabies virus phophoprotein P, Rabies virus matrix protein M, Rabies virus glycoprotein G, RSV fusionprotein F, RSV nucleoprotein N, RSV matrix protein M, RSV matrix protein M2-1, RSV matrix protein M2-2, RSV phophoprotein P, RSV small hydrophobic protein SH, RSV major surface glycoprotein G, RSV polymerase L, RSV non-structural protein 1 NS1, RSV non-structural protein 2 NS2, Staphylococcus secretory antigen SssA, Yellow fever virus genome polyprotein, Yellow fever virus protein E, Yellow fever virus protein M, Yellow fever virus capsid protein C, Yellow fever virus protease NS3, Yellow fever virus protein NS1, Yellow fever virus protein NS2A, Yellow fever virus protein AS2B, Yellow fever virus protein NS4A, Yellow fever virus protein NS4B, and Yellow fever virus protein NS5.

7. The nucleic acid molecule of claim 1 , wherein the molecule does not comprise sequence encoding a reporter protein, a marker or selection protein.

8. The nucleic acid molecule of claim 1 , wherein the nucleic acid is an RNA.

9. The nucleic acid molecule of claim 1 , wherein the poly(A) sequence comprises a sequence of about 25 to about 400 adenosine nucleotides.

10. The nucleic acid molecule of claim 1 , wherein the polyadenylation signal comprises the consensus sequence NN(U/T)ANA, preferably AA(U/T)AAA or A(U/T)(U/T)AAA.

11. The nucleic acid molecule of claim 1 , wherein at least one guanosine, uridine, adenosine, thymidine, or cytidine

Assignees

• Curevac Gmbh
• Curevac Ag

Inventors

• Thess Andreas
• Schlake Thomas
• Probst Jochen

Classifications

• A61P33/00

  Antiparasitic agents · CPC title

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• A61P31/04

  Antibacterial agents · CPC title

• A61P31/10

  Antimycotics · CPC title

• A61P31/12

  Antivirals · CPC title