ARENAVIRES FOR USE IN THE TREATMENT OF TUMORS IN HUMANS

Description

FIELD OF APPLICATION AND STATE OF THE ART

[0001] The invention relates to arenaviruses for use in the treatment of tumors as defined in the claims.

[0002] Arenaviruses belong to the family of human-pathogenic, pleomorphic RNA viruses. Diseases caused by these viruses are classified as zoonoses due to their natural reservoir in animals, predominantly rodents. Zoonoses are diseases that can be transmitted from animals to humans and vice versa.

[0003] At least eight arenaviruses are known to cause disease in humans. Typical causes include aseptic meningitis and hemorrhagic fever. Known viruses that can cause disease in humans include lymphocytic choriomeningitis virus (LCMV), Guanarito virus (GTOV), Junin virus (JUNV), Lassa virus (LASV), Lujo virus (LUJV), Machupo virus (MACV), Sabia virus (SABV), and Whitewater Arroyo virus (WWAV).

[0004] Arenaviruses are generally divided into two groups: Old World arenaviruses and New World arenaviruses. These groups differ geographically and genetically. Old World arenaviruses, such as lymphocytic choriomeningitis virus, have been found in countries of the Eastern Hemisphere, such as Europe, Asia, and Africa. New World arenaviruses, on the other hand, have been found in countries of the Western Hemisphere, such as Argentina, Bolivia, Venezuela, Brazil, and the United States.

[0005] The name of the virus family is derived from the Latin arenosus (sandy) or arena (sand), describing the sandy ribosomal structure inside the virions. Arenavirus virions have a round to irregular shape and, depending on the species and preparation of the specimen, range in diameter from 50 nm to 300 nm, usually between 110 nm and 130 nm. Embedded in the viral envelope are 8 nm to 10 nm long, club-shaped glycoprotein spikes. The individual spikes consist of a tetramer of the viral envelope protein.

[0006] The virions also contain two circular capsids with helical symmetry. The length of the capsids varies between 450 nm and 1300 nm. Each capsid contains a molecule of viral RNA (ribonucleic acid) polymerase (L protein).

[0007] Each capsid contains a molecule of single-stranded RNA with mixed (i.e., ambisense, + / -) polarity. The two single-stranded RNA molecules constitute the viral genome. They are designated L (large) and S (small) and are approximately 7.5 kb (kilobases) and 3.5 kb (kilobases), respectively. Although the capsids are closed in a ring, the RNA strands are linear and therefore not circular. A 19- to 30-base-long sequence at the 3' end of the RNA is present on both strands and is also conserved within the virus family.

[0008] Morphologically quite unusual is the presence of a varying number of cellular ribosomes within the virions, which give the virus particles their "sandy" appearance. Likewise, purified virus preparations contain a number of different cellular RNAs (including ribosomal RNA), as well as replicative forms of viral RNA, such as various viral mRNAs (messenger ribonucleic acids bound to the ribosomes) and complete complementary strands of the viral genome. These non-genomic RNAs are all found in varying amounts outside the aforementioned capsids.

[0009] The use of arenaviruses as vaccine vectors is well known. A prominent example is the Candid #1 vaccine virus used against Argentine hemorrhagic fever. This is a vaccine variant of the Junin virus.

[0010] WO 2009 / 083210 A1 discloses the use of replication-defective, genetically modified, arenavirus particles (virions) for the treatment of neoplastic diseases such as melanoma, prostate cancer, breast cancer, and lung cancer, among others. The publication "Development of replication-defective lymphocytic choriomeningitis virus vectors for the induction of potent CD8+ T cell immunity" (Nature Medicine, Vol. 16, No. 3, March 2010, pp. 339-345; doi: 10.1038 / nm.2104) mentions cancer immunotherapy as a potential application for such virus particles.

[0011] Furthermore, WO 2006 / 008074 A1 discloses the use of packaging cells producing retroviral virions pseudotyped with arenavirus glycoprotein for gene therapy of solid tumors.

[0012] Kelly, E. & Russell, S. (2007) Molecular Therapy, Vol. 15, No. 4, 651–659 describes the history of oncolytic viruses: from origin to genetic engineering. Molomut, N. & Padnos, M. (1965) Nature, Vol. 208, No. 5014, 948–950 reveals the inhibition of transplantable and spontaneous mouse tumors by the MP virus. Reiserova, L. et al. (1999) Virology, Vol. 257, No. 1, 73–83 reveals the identification of the MaTu-MX pathogen as a new strain of lymphocytic choriomeningitis virus (LCMV) and serological evidence for horizontal spread of LCMV in the human population. Koma, T. et al. (2013) J. Mol. Biol., Vol. 425, No. 24, pp. 4893–4903 describes the innate immune response to arenavirus infections, focusing on the highly pathogenic hemorrhagic New World arenaviruses. Groseth, A. et al. (2011) PLOS Neglected tropical diseases, Vol. 5, No.5, e1137 reveals that infection with Tacaribe virus, but not with Junin virus, induces the release of cytokines from primary human monocytes and macrophages. Webb, HE et al. (1975) Clinical Oncology 1975, Vol. 1, No. 2, 157–169 discloses the treatment of 18 cases of malignant disease with an arenavirus. Schadler, Keri L. et al. (2014) Cancer research, Vol. 74, No. 8, 2171–2181 discloses immunosurveillance through antiangiogenesis, particularly tumor growth arrest by T cell-produced thrombospondin-1.

[0013] The methods described in the state of the art for treating tumors are based on the use of virus particles that are very complex to produce using genetic engineering. In addition, gene therapy treatment methods often fail to achieve sufficient, therapeutically effective transduction of tumor tissue with genetically engineered virions or packaging cells that produce virions. TASK AND SOLUTION

[0014] The present invention is therefore based on the object of providing a simpler and, in particular, more efficient therapy solution for tumors, in particular carcinomas and sarcomas, compared to the prior art.

[0015] This object is achieved according to the invention by an arenavirus for use according to independent claim 1. Preferred embodiments are defined in the dependent claims.

[0016] According to a first aspect, the invention relates to an arenavirus for use in the treatment of a tumor, preferably a malignant, i.e., malignant, tumor in humans, whereby tumor regression is effected, wherein the arenavirus is free of foreign genomic RNA, wherein the arenavirus is lymphocytic choriomeningitis virus, WE strain.

[0017] The arenavirus is characterized by its lack of foreign genomic RNA, meaning it contains no foreign genomic RNA. In other words, the genome (genetic material) of the arenavirus is free of foreign RNA, or preferably contains no foreign RNA.

[0018] For the purposes of the present invention, the term "genomic foreign RNA" refers to an RNA (ribonucleic acid) or RNA sequence that does not occur or is not contained in the genome of a wild-type arenavirus or in the genome of a mutant of a wild-type arenavirus (mutated arenavirus), in particular in the genome of a natural mutant of a wild-type arenavirus (naturally mutated arenavirus). Examples of foreign RNA include artificial or synthetic RNA molecules, RNA from organisms, and RNA from other viruses.

[0019] For the purposes of the present invention, the term "wild-type arenavirus" is understood (in accordance with the understanding of the person skilled in the art) to mean an arenavirus whose genome represents the normal genetic form occurring in nature.

[0020] The term "mutant of a wild-type arenavirus" or "mutated arenavirus" is understood in the context of the present invention (in accordance with the understanding of the person skilled in the art) to mean an arenavirus whose genome exhibits a spontaneous, i.e. naturally caused, or mutagen-induced change compared to the wild-type genome.

[0021] Accordingly, the term "natural mutant of a wild-type arenavirus" or "naturally mutated arenavirus" within the meaning of the present invention (in accordance with the understanding of the person skilled in the art) is understood to mean an arenavirus whose genome exhibits a spontaneous, i.e., naturally induced, change compared to the wild-type genome. A naturally mutated arenavirus can preferably be produced by passage, particularly serial passage, which will be discussed in more detail below.

[0022] The invention is based on the surprising finding that arenaviruses without foreign genomic RNA are capable of inducing tumor regression. Tumor regression is based on the activation or stimulation of innate and adaptive immune cells caused by the arenaviruses. The activated immune cells secrete increased amounts of antitumor cytokines such as interferon-α and interferon-γ, thereby fighting or repelling the tumor. Another surprising finding is the realization that, in the event of tumor manifestation, arenaviruses cause a significantly increased secretion of antitumor cytokines. Arenaviruses without foreign genomic RNA are therefore suitable for use in tumor treatment. This has been successfully verified by the applicant in animal experiments. Among other things, mice, in which human tumor growth is possible, were used for this purpose.

[0023] The arenavirus is free of non-genomic foreign RNA. For the purposes of the present invention, the term "non-genomic foreign RNA" refers to an RNA or RNA sequence outside the arenavirus genome that does not occur or is not contained in a wild-type arenavirus or a mutant of a wild-type arenavirus (mutated arenavirus), in particular a natural mutant of a wild-type arenavirus (naturally mutated arenavirus). In other words, in this embodiment, the arenavirus contains no foreign RNA at all, i.e., neither genomic foreign RNA nor non-genomic foreign RNA.

[0024] For the purposes of the present invention, the term "passage" is understood to mean, in accordance with the expert's understanding, the multiple, regular introduction of the arenavirus into host animals and / or host cells. For the purposes of the present invention, the term "serial passage" is understood to mean, in accordance with the expert's understanding, the multiple, regular introduction of the arenavirus into different host animals, preferably of the same species, and / or different cells, preferably of the same type. Due to the multiple changes in environment (host animal and / or host cell), the arenavirus is exposed to increased adaptation pressure or mutation pressure, thereby increasing the probability of mutations in the arenavirus genome that are advantageous from a tumor regression perspective.

[0025] In one embodiment, the tumor is selected from the group comprising or consisting of carcinoma, melanoma, blastoma, lymphoma, and sarcoma.

[0026] For the purposes of the present invention, the term "carcinoma" is understood to mean (in accordance with the understanding of the person skilled in the art) a malignant neoplasm of epithelial origin.

[0027] For the purposes of the present invention, the term "sarcoma" is understood (in accordance with the understanding of the person skilled in the art) to mean a malignant neoplasm of mesodermal origin.

[0028] For the purposes of the present invention, the term "melanoma" is to be understood (in accordance with the understanding of the person skilled in the art) as a malignant neoplasm of melanocytic origin.

[0029] For the purposes of the present invention, the term "lymphoma" is understood (in accordance with the understanding of the person skilled in the art) to mean a malignant neoplasm of lymphocytic origin.

[0030] For the purposes of the present invention, the term "blastoma" is understood (in accordance with the understanding of the person skilled in the art) to mean a malignant neoplasm of embryonic origin.

[0031] In a preferred embodiment, the carcinoma is selected from the group comprising or consisting of anal carcinoma, bronchial carcinoma, lung carcinoma, endometrial carcinoma, gallbladder carcinoma, hepatocellular carcinoma, testicular carcinoma, colorectal carcinoma, laryngeal carcinoma, esophageal cancer, gastric carcinoma, breast carcinoma, kidney carcinoma, ovarian carcinoma, pancreatic tumor, pharyngeal carcinoma, prostate carcinoma, thyroid carcinoma and cervical carcinoma.

[0032] In a preferred embodiment, the sarcoma is selected from the group comprising or consisting of angiosarcoma, chondrosarcoma, Ewing's sarcoma, fibrosarcoma, Kaposi's sarcoma, liposarcoma, leiomyosarcoma, malignant fibrous histiocytoma, neurogenic sarcoma, osteosarcoma and rhabdomyosarcoma.

[0033] The arenavirus is the lymphocytic choriomeningitis virus (LCMV), WE strain.

[0034] The arenavirus can be delivered in the form of virions, i.e. in the form of arenavirus particles located outside a cell.

[0035] Alternatively, the arenavirus is intended, preferably prepared, for local, particularly intramuscular, intraperitoneal, or subcutaneous, administration. In other words, the arenavirus is used for local, particularly intramuscular, intraperitoneal, or subcutaneous, administration.

[0036] The arenavirus is preferably intended, preferably prepared, for local administration in a dose of 1 PFU (plaque forming unit) / kg body weight to 10 12< PFU / kg body weight, in particular 10 2< PFU / kg body weight to 10 6< PFU / kg body weight, preferably 10 3< PFU / kg body weight to 10 5< PFU / kg body weight. In other words, the arenavirus is preferably used for local administration in a dose of 1 PFU (plaque forming unit) / kg body weight to 10 12< PFU / kg body weight, in particular 10 2< PFU / kg body weight to 10 6< PFU / kg body weight, preferably 10 3< PFU / kg body weight to 10 5< PFU / kg body weight.

[0037] Alternatively, the arenavirus is intended, preferably prepared, for systemic, particularly intravenous, administration. In other words, in an alternative embodiment, the arenavirus is used for systemic, particularly intravenous, administration.

[0038] Preferably, the arenavirus is intended, preferably prepared, for systemic administration in a dose of 1 PFU / kg body weight to 10 12< PFU / kg body weight, in particular 10 2< PFU / kg body weight to 10 6< PFU / kg body weight, preferably 10 3< PFU / kg body weight to 10 5< PFU / kg body weight. In other words, the arenavirus is preferably used for systemic administration in a dose of 1 PFU / kg body weight to 10 12< PFU / kg body weight, in particular 10 2< PFU / kg body weight to 10 6< PFU / kg body weight, preferably 10 3< PFU / kg body weight to 10 5< PFU / kg body weight.

[0039] According to a second aspect, the invention relates to a medicament for use in the treatment of a tumor in humans, in particular a malignant tumor.

[0040] The drug is characterized by the fact that it contains an arenavirus as described above.

[0041] The medicament preferably further comprises a pharmaceutically acceptable carrier. The carrier can in particular be selected from the group comprising or consisting of water, saline solution, buffer solution, and cell culture medium.

[0042] The medicament also contains an active ingredient. The active ingredient can be, in particular, a cytostatic agent, an antibody, and / or a cytokine.

[0043] Regarding further features and advantages of the drug, in particular the arenavirus and the tumor, reference is made entirely to the previous description to avoid repetition.

[0044] Further features and advantages of the invention will become apparent from the following description of preferred embodiments in the form of exemplary embodiments, the associated figures and the claims. Example part 1. Methods and materials 1. 1 mice

[0045] Unless otherwise stated, the mice used were on a C57BL / 6 background. Map3k14 aly / aly < Mice lack NF-kB signals and are thus severely immunosuppressed. Irf3 × Ir7 - / - < Mice cannot produce interferon. NOD.SCID mice lack an adaptive immune system. Thus, human tumor growth is possible in these mice. LoxP-tagged mice spontaneously develop liver tumors. 1.2 Cell lines and reagents

[0046] MOPC cells are murine oropharyngeal carcinoma cells. Mc38 are murine colon carcinoma cells. Raw cells are immortalized macrophages. A431 are human lung carcinoma cells; Sw40 are human colon carcinoma cells, and HeLa are human cervical carcinoma cells. Primary macrophages were cultured from bone marrow progenitor cells using M-CSF. Cells were maintained in Dulbecco's modified Eagle's medium with 10% fetal bovine serum (Sigma-Aldrich), 2 mmol / I L-glutamine, and 100 U / ml penicillin. All cells were cultured in 5% CO2. 1.3 Viruses

[0047] The LCMV strain WE was obtained from the laboratory of Prof. Zinkernagel (Experimental Immunology, Zurich, Switzerland) and propagated in L929 cells. Candid#1 was obtained from Professor Paula Cannon (University of Southern California). 1.4 Tumor growth and treatments

[0048] Approximately 5 × 10 5 tumor cells (in 100 microL) were injected subcutaneously into the right flank of 6- to 12-week-old mice. The longest tumor diameter was measured. Mice were treated with peritumoral injections of 2 × 10 4 PFU of LCMV-WE or Candid#1 (in 100-200 microL). 1.5 Morphometric analysis of tumor vessels

[0049] Morphometric analyses were performed on consecutive frozen sections stained for the endothelial cell marker CD31. Vascular density (MVD) was calculated using the average of three tumor sections. MVD was calculated as the number of vessels per tumor area. 1.6 Detection of hypoxia

[0050] Hypoxic tumor areas were detected by the formation of pimonidazole adducts after injection of pimonidazole into tumor-transplanted animals for 30 min. Tumor sections were stained using the Hypoxyprobe-1 Plus kit according to the manufacturer's instructions (Pharmacia Natur International, Inc.). 1.7 IFN-α ELISA

[0051] Serum IFN-α levels were determined by ELISA according to the manufacturer's instructions (Research Diagnostics RDI, Flanders, NJ). 1.8 Statistical analysis

[0052] Means were compared using an unpaired Student's two-tailed t-test. Data are presented as mean ± SEM. The level of statistical significance was set at p < 0.05. 2. Investigations

[0053] 2.1Immortalized macrophages (tumor cells) and primary bone marrow-derived macrophages (primary) were infected with LCMV (strain WE). Replication was measured after 24 hours (n = 3).

[0054] It was demonstrated that LCMV (strain WE) replicates in both immortalized and healthy cells. The results are presented in Figure 1 shown graphically.

[0055] The Figure 1 has the following legend: Ordinate: LCMV (log 10 PFU / ml) Abscissa: Tumor cells / healthy macrophages (primary)

[0056] 2.2 WT C57BL / 6 mice were treated with 5 × 10 5 MOPC cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 PFU LCMV (strain WE) (day 0). Tumor growth was monitored.

[0057] It was demonstrated that treatment with LCMV resulted in almost complete tumor regression. The results are presented in Figure 2 shown graphically.

[0058] The Figure 2 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0059] 2.3 WT C57BL / 6 mice were treated with 5 × 10 5 < MC38 cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 < PFU LCMV (strain WE) (day 0). Tumor growth was monitored.

[0060] It was demonstrated that treatment with LCMV resulted in significant tumor regression. The results are presented in Figure 3 shown graphically.

[0061] The Figure 3 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0062] 2.4Approximately nine-month-old LoxP-TAg mice with spontaneously developing liver carcinomas were infected intravenously with 2 × 10 6 < PFU LCMV or left untreated. Tumor nodules (diameter >=3 mm) were macroscopically quantified on day 6 (n = 3) and day 20 (n = 4-5).

[0063] It was demonstrated that treatment with LCMV significantly reduced the number of carcinomatous liver nodules. The results are presented in Figure 4 shown graphically.

[0064] The Figure 4 has the following legend: Ordinate: Liver nodules (number) Abscissa: Time

[0065] 2.5 WT C57BL / 6 mice (n = 4 per group) were injected subcutaneously with 5 × 10 5 MOPC cells (day -3) or LCMV (strain WE) 2 × 10 4 PFU (day 0), or both 5 × 10 5 MOPC cells (day -3) and 2 × 10 4 PFU LCMV (day 0). Serum samples were collected on day 3, and an IFN-α ELISA was performed.

[0066] It was demonstrated that LCMV caused a drastically increased secretion of interferon-γ in experimental animals that were simultaneously administered carcinoma cells. The results obtained are presented in Figure 5 shown graphically.

[0067] The Figure 5 has the following legend: Ordinate: IFN-α (pg / ml)

[0068] 2.6 Map3k14 aly / aly < mice and WT mice were treated with 5 × 10 5 < MOPC cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 < PFU LCMV (strain WE) (day 0). Tumor growth was monitored.

[0069] It was demonstrated that treatment with LCMV resulted in tumor regression. The results are presented in Figure 6 shown graphically.

[0070] The Figure 6 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0071] 2.7 Irf3 × Ir7 - / - < mice and WT mice were treated with 5 × 10 5 < MOPC cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 < PFU LCMV (strain WE) (day 0). Tumor growth was monitored.

[0072] It was demonstrated that treatment with LCMV resulted in tumor regression. The results are presented in Figure 7 shown graphically.

[0073] The Figure 7 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0074] 2.8 WT mice were treated with 5 × 10 5 MOPC cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 PFU LCMV (strain WE) (day 0). On day 9 after tumor administration, the tumors were analyzed histologically using CD31 staining. Vascular density (MVD) and vessel-to-vessel distance were quantified.

[0075] It was demonstrated that treatment with LCMV resulted in a reduction in tumor vessel density. The results are presented in Figure 8A shown graphically.

[0076] The Figure 8A has the following legends: Left side: Right side: Ordinate: MVD [mm 3< ] Ordinate: vessel-vessel distance (µm) Abscissa: Tumor / Tumor LCMV Abscissa: Tumor / Tumor LCMV

[0077] 2.9 WT mice were treated with 5 × 10 5 MOPC cells (day 3). One group of mice was additionally treated peritumorally with 2 × 10 4 PFU LCMV (strain WE) (day 0). On day 9, the animals were injected with pimonidazole, and the tumors were subsequently analyzed histologically for hypoxic areas.

[0078] It was demonstrated that treatment with LCMV caused a lack of oxygen in the carcinoma tissue. The results are presented in Figure 8B shown graphically.

[0079] The Figure 8B has the following legend: Ordinate: Hypoxic areas / Tumor (%) Abscissa: Tumor / Tumor LCMV

[0080] 2.10 WT C57BL / 6 mice were injected subcutaneously with 5 × 10 5 MOPC cells in the right flank (day 3). On day 0, one group of animals was treated with 2 × 10 4 PFU LCMV (strain WE) in the right flank (ipsilateral), left flank (contralateral), or intravenously. Tumor growth was monitored.

[0081] It was demonstrated that treatment with LCMV resulted in tumor regression even with systemic administration. The results are presented in Figure 9 shown graphically.

[0082] The Figure 9 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0083] 2.11NOD.SCID mice were injected subcutaneously with 5 × 10 5 < A431 cells (day -3) and then either left untreated or treated with 2 × 10 4 < PFU LCMV (strain WE). Tumor size (longest diameter) was measured on the indicated day. Mice were sacrificed when the tumor size reached 12 mm.

[0084] It was demonstrated that treatment with LCMV increased the survival rate in the test animals. The results are presented in Figure 10 shown graphically.

[0085] The Figure 10 has the following legend: Ordinate: Survival (%) Abscissa: Time (days)

[0086] 2.12 NOD.SCID mice were treated with 5 × 10 5 < Sw40 cells (day 0). One group of mice was additionally treated peritumorally with 2 × 10 4 < PFU LCMV (strain WE) or 2 × 10 4 < PFU Candid#1 (day 0). Tumor growth was monitored.

[0087] It was demonstrated that treatment with LCMV and Candid#1 resulted in tumor regression. The results are presented in Figure 11 shown graphically.

[0088] The Figure 11 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0089] 2.13 NOD.SCID mice were treated with 5 × 10 5 Hela cells (day 0). One group of mice was additionally treated peritumorally with 2 × 10 4 PFU LCMV (strain WE) (day 3). Tumor growth was monitored.

[0090] It was demonstrated that treatment with LCMV resulted in tumor regression. The results are presented in Figure 12 shown graphically.

[0091] The Figure 12 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0092] 2.14NOD.SCID mice were treated with 5 × 10 5 < HepG2 cells (day -10) and then additionally treated peritumorally with or without 2 × 10 4 < PFU of Candid #1 (day 0). Tumor growth was monitored.

[0093] It was demonstrated that treatment with Candid#1 resulted in tumor regression in this tumor type. The results are presented in Figure 13 shown graphically.

[0094] The Figure 13 has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0095] 2.15 Primary human cells (hepatocytes, colon epithelial cells, melanocytes) and tumor cells from the same tissue of origin were infected with LCMV (MOl 1). Virus levels were measured in the supernatant after 1, 2, and 3 days.

[0096] In this experiment, it was shown that arenaviruses replicate more frequently in tumor cells than in healthy tissue.

[0097] The Figure 14has the following legend: Ordinate: Infectious virus in cell culture supernatant (logarithmized plaque forming units) Abscissa: Time (days)

[0098] 2.16 Tumor diameter (A) and survival (B) of C57BL / 6 mice bearing one metastasis in the shoulder and one metastasis in the flank (MOPC cells) that were left untreated or treated with 2×10 6< PFU LCMV intravenously.

[0099] In this experiment, it was shown that intravenous therapy of LCMV is very effective against two local metastases and thus prolongs survival.

[0100] The Figure 15 A has the following legend: Ordinate: Tumor diameter of both metastases (cm) Abscissa: Time (days)

[0101] The Figure 15 B has the following legend: Ordinate: Survival in percent Abscissa: Time (days)

[0102] 2.17Tumor diameter of C57BL / 6 mice bearing melanoma (B16F10 cells) left untreated or treated with 2×10 4< PFU LCMV intratumorally.

[0103] This experiment showed that local therapy with LCMV is very effective in treating melanoma.

[0104] The Figure 16 has the following legend: Ordinate: Tumor diameter of the melanoma (cm) Abscissa: Time (days)

[0105] 2.18 Number of melanomas from MT / ret mice (developing endogenous melanomas) left untreated or treated with 2×10 6< PFU LCMV intravenously.

[0106] This experiment showed that systemic therapy with LCMV is very effective in treating melanoma.

[0107] The Figure 17 has the following legend: Ordinate: Number of melanomas

[0108] 2.19Tumor diameter (A) and survival (B) of NOD.SCID mice bearing human fibrosarcoma (Sw872 cells) left untreated or treated with 2×10 6< PFU Candid#1 intratumorally.

[0109] In this experiment it was shown that Candid#1 is also very effective in fibrosarcoma and thus prolongs survival.

[0110] The Figure 18 A has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0111] The Figure 18 B has the following legend: Ordinate: Survival in percent Abscissa: Time (days)

[0112] 2.20 Tumor diameter (A) and survival (B) of NOD.SCID mice bearing human pharyngeal carcinoma (FaDu cells) that were left untreated or treated with 2×10 6< PFU LCMV intratumorally.

[0113] This experiment showed that LCMV is also very effective in human pharyngeal carcinoma and thus prolongs survival.

[0114] The Figure 19 A has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0115] The Figure 19 B has the following legend: Ordinate: Survival in percent Abscissa: Time (days)

[0116] 2.21 Expression of receptors on tumor-specific T cells (PD-1, IL2 receptor, IL7 receptor) that influence T cell function. Tumor-specific T cells were obtained from the blood of mice with B16F10 tumors that were additionally treated intratumorally with or without LCMV.

[0117] In this experiment it was shown that LCMV has a positive effect on tumor-specific T cells.

[0118] The Figure 20has the following legend: Ordinate: Strength of expression of the different receptors (Mean fluorescence intensity)

[0119] 2.22 Tumor diameter (A) and survival (B) of C57BL / 6 mice bearing murine subcutaneous lymphoma (EL4 cells) treated with or without tumor-specific T cells (OT1 cells) and additionally treated with or without LCMV intratumorally (2×10 6< PFU).

[0120] This experiment showed that LCMV works synergistically with T cell therapy.

[0121] The Figure 21 A has the following legend: Ordinate: Tumor diameter (cm) Abscissa: Time (days)

[0122] The Figure 21 B has the following legend: Ordinate: Survival in percent Abscissa: Time (days)

[0123] 2.23 Survival of C57BU6 mice and PD-1 deficient mice ( Pdcd1- / -< mice) bearing murine pharyngeal carcinoma (MOPC cells) and treated intratumorally with LCMV (2×10 4< PFU).

[0124] In this experiment it was shown that LCMV acts synergistically with PD-1 blockade.

[0125] The Figure 22 has the following legend: Ordinate: Survival in percent Abscissa: Time (days)

[0126] The nucleic acid sequences SEQ ID No. 1 to SEQ ID No. 6 mentioned in the general description correspond to the nucleic acid sequences disclosed in the following sequence listing. Sequence protocol (SEQUENCE LISTING)

[0127] <110> Lang Karl Sebastian <120> Arenaviruses for use in the treatment of carcinomas or sarcomas and processes for the production of arenaviruses with improved immunotherapeutic effects against carcinomas or sarcomas <130> P54975WO <160> 6 <170> BiSSAP 1.3 <210> 1 <211> 3413 <212> RNA <213> Junin virus <220> <223> Junin virus strain Candid-1 segment S - AY746353 - viruses; ssrna negative-strand viruses; arenaviridae; arenavirus; new world arenaviruses <400> 1 <210> 2 <211> 7114 <212> RNA <213> Junin virus <220> <223> Junin virus strain Candid #1 segment L - AY819707 - viruses; ssrna negative-strand viruses; arenaviridae; arenavirus; new world arenaviruses <400> 2 <210> 3 <211> 3410 <212> RNA <213> Junin mammarenavirus <220> <223> Junin virus strain Candid #1 segment S, completesequence.-ACCESSION FJ969442-VERSION FJ969442.1-GI:264665554 - Viruses; ssRNA viruses; ssRNA negative-strand viruses; Arenaviridae; Mammarenavirus. <400> 3 <210> 4 <211> 7114 <212> RNA <213> Junin mammarenavirus <220> <223> Junin virus strain Candid #1 segment L, complete sequence.-ACCESSION AY819707-VERSION AY819707.2-GI:227957900-Viruses; ssRNA viruses; ssRNA negative-strand viruses; Arenaviridae; Mammarenavirus. <400> 4 <210> 5 <211> 3377 <212> RNA <213> Lymphocytic choriomeningitis mammarenavirus Viruses (LCMV) <220> <223> Lymphocytic choriomeningitis virus clone 13 segment S, complete sequence.ACCESSION DQ361065-VERSION DQ361065.2-GI:116563461.-ssRNA viruses; ssRNA negative-strand viruses; Arenaviridae; Mammarenavirus. <400> 5 <210> 6 <211> 7229 <212> RNA <213> Lymphocytic choriomeningitis mammarenavirus Viruses (LCMV) <220> <223> Lymphocytic choriomeningitis virus clone 13 segment L, complete sequence.-ACCESSION DQ361066-VERSION DQ361066.1-GI:86440167.-ssRNA viruses; ssRNA negative-strand viruses; Arenaviridae; Mammarenavirus <400> 6

Claims

1. An arenavirus for use in the treatment of a tumor in a human, wherein tumor regression is effected, wherein the arenavirus is free of foreign genomic RNA, wherein the arenavirus is lymphocytic choriomeningitis virus, WE strain.

2. The arenavirus for the use of claim 1, wherein the treatment comprises activating innate and adaptive immune cells, thereby causing tumor regression.

3. The arenavirus for the use of claim 1 or 2, wherein the tumor is a carcinoma, melanoma, blastoma, lymphoma or sarcoma.

4. A medicament for use in the treatment of a tumor, in a human, wherein the medicament comprises an arenavirus of any one of the preceding claims and preferably a pharmaceutically acceptable carrier.