Use of baloxavir sodium in treatment of crimean-congo hemorrhagic fever virus infection

Abstract

The present application relates to use of baloxavir sodium represented by formula (I) and / or a solvate thereof and / or a hydrate thereof in the preparation of a drug for the treatment of bunyavirus infection, particularly Crimean-Congo hemorrhagic fever virus (CCHFV) infection.

Description

Use of sodium baloxavir for treating Crimean-Congo hemorrhagic fever virus infection

[0001] Cross-reference to Related Applications

[0002] This application is based on and claims priority to CN application No. 202410479652.1, filed on April 19, 2024, the disclosure of which is incorporated herein in its entirety. TECHNICAL FIELD

[0003] The present application relates to the use of sodium baloxavir and / or solvate and / or hydrate thereof represented by the following formula I in the preparation of a medicament for treating Bunyavirus, in particular Crimean-Congo hemorrhagic fever virus (CCHFV) infection, and to the use of a composition, such as an injection, comprising the compound and / or solvate and / or hydrate thereof in the preparation of a medicament for treating Bunyavirus, in particular Crimean-Congo hemorrhagic fever virus (CCHFV) infection. BACKGROUND

[0004] Crimean-Congo hemorrhagic fever (CCHF) is an acute infectious disease widely prevalent in Africa, Asia and Europe, etc., with fever, gastrointestinal bleeding, hematemesis, shock as the main characteristics, and the mortality rate can reach 50%. The pathogen of CCHF is Crimean-Congo hemorrhagic fever virus (CCHFV). Crimean-Congo hemorrhagic fever virus belongs to Bunyavirales, Nairoviridae, Orthonairovirus tick-borne virus. The clinical symptoms of CCHF patients are extensive, and the typical course of disease experiences four stages: incubation period, pre-bleeding period, bleeding period and recovery period. The length of the incubation period is related to factors such as contact route and viral load; after the incubation period, the pre-bleeding stage shows fever disease, and patients have severe headache, nausea, diarrhea, muscle soreness, photophobia and other symptoms. When patients enter the bleeding stage, the body will appear petechiae, large areas of ecchymosis and massive bleeding in turn, and 9% to 50% of cases die in this stage. In addition to these manifestations, studies have found that CCHF is accompanied by severe liver, nervous system, respiratory system and heart damage and other lesions at the same time of onset. There is no marketed drug and vaccine against Crimean-Congo hemorrhagic fever. SUMMARY

[0005] The application aims to find a drug with antiviral activity against Bunyaviruses, especially Crimean-Congo hemorrhagic fever virus, which can be used for the treatment of diseases caused by its infection, such as fever, gastrointestinal bleeding, hematemesis, shock, etc. Baloxavir acid has in vitro activity against Crimean-Congo hemorrhagic fever virus, but its solubility is low and cannot be administered by injection. Through creative research, the application finds that the compound baloxavir sodium shown in formula I has good solubility and has in vivo and in vitro activity in inhibiting Crimean-Congo hemorrhagic fever virus, and can be used for preparing a pharmaceutical injection, and has a good therapeutic effect in treating diseases caused by Crimean-Congo hemorrhagic fever virus.

[0006] The application provides a compound having the structure of formula I and / or a solvate thereof and / or a hydrate thereof:

[0007] The compound shown in formula I can inhibit viral replication on cells and reduce viral nucleic acid load in cell culture.

[0008] The inventors of the application found some new action characteristics of the compound of formula I after creative invention and research:

[0009] First, the compound shown in formula I can reduce the viral nucleic acid load level of Crimean-Congo hemorrhagic fever infected cells at a micromolar concentration;

[0010] Second, the compound shown in formula I has a significant protective effect on mice infected with Crimean-Congo hemorrhagic fever virus (CCHFV).

[0011] The application relates to the use of the compound shown in formula I and / or a solvate thereof and / or a hydrate thereof in the preparation of a drug for treating a disease or infection caused by a Bunyavirus in a subject,

[0012] The application also relates to the use of the compound shown in formula I and / or a solvate thereof and / or a hydrate thereof in the preparation of a drug as a Bunyavirus inhibitor.

[0013] The application also relates to the use of the compound shown in formula I and / or a solvate thereof and / or a hydrate thereof in the preparation of a drug for inhibiting the replication or proliferation of a Bunyavirus in cells (such as mammalian cells) in vivo or in vitro.

[0014] The application also relates to the use of the composition in the preparation of a drug for treating a disease or infection caused by a Bunyavirus in a subject.

[0015] The application also relates to the use of the composition in the preparation of a drug as a Bunyavirus inhibitor.

[0016] The present application also relates to the use of the composition in the preparation of a medicament for inhibiting the replication or propagation of a Bunyavirus in a cell (e.g., a mammalian cell) in vivo or in vitro.

[0017] The present application also relates to a method for treating and / or preventing a disease or infection in a subject in need thereof, comprising administering to the subject in need thereof a therapeutically and / or prophylactically effective amount of the composition comprising the compound of Formula I and / or solvate thereof and / or hydrate thereof or the compound of Formula I and / or solvate thereof and / or hydrate thereof, wherein the disease comprises a disease or infection caused by a Bunyavirus.

[0018] The present application also relates to a method for inhibiting the replication or propagation of a Bunyavirus in a subject in need thereof, comprising administering to the subject in need thereof a therapeutically and / or prophylactically effective amount of the composition comprising the compound of Formula I and / or solvate thereof and / or hydrate thereof or the compound of Formula I and / or solvate thereof and / or hydrate thereof.

[0019] In certain embodiments, the composition comprises the compound of Formula I and / or solvate thereof and / or hydrate thereof. In certain embodiments, the compound of Formula I and / or solvate thereof and / or hydrate thereof in the composition is an active ingredient. In certain embodiments, the compound of Formula I and / or solvate thereof and / or hydrate thereof in the composition is present in an effective amount. In certain embodiments, the composition further comprises a pharmaceutically acceptable carrier or excipient. In certain embodiments, the composition is an injection.

[0020] In certain embodiments, the injection comprises:

[0021] (1) the compound of Formula I and / or solvate thereof and / or hydrate thereof in an amount of 0.1-20%;

[0022] (2) a solvent selected from the group consisting of water for injection, physiological saline, sterile water, and the like, in an amount of 60-99%;

[0023] (3) one or more excipients selected from the group consisting of:

[0024] a pH adjusting agent such as hydrochloric acid, acetic acid, and the like, in an amount of 0.01-5%;

[0025] an isotonic agent such as mannitol, sodium chloride, and the like, in an amount of 0.1-10%;

[0026] a preservative such as p-hydroxybenzoic acid ester, and the like, in an amount of 0.01-1%;

[0027] a stabilizer such as ethanol, propylene glycol, and the like, in an amount of 0.1-10%.

[0028] In certain embodiments, the pH of the injection is between 4 and 9.

[0029] The present application also relates to the use of the injection for the preparation of a medicament for the treatment of a disease or infection caused by a Bunyavirus in a subject.

[0030] The present application also relates to the use of the injection for the preparation of a medicament for the inhibition of the replication or the propagation of a Bunyavirus in a cell, e.g. a mammalian cell, in vivo or in vitro.

[0031] The present application also relates to the use of the injection for the preparation of a medicament as a Bunyavirus inhibitor.

[0032] In certain embodiments, the compound of formula I and / or solvates and / or hydrates thereof are administered to a subject by injection as an active ingredient.

[0033] In certain embodiments, the compound of formula I and / or solvates and / or hydrates thereof or the composition or the medicament is preferably administered to a subject by injection, e.g. intravenously, intramuscularly or by other injection.

[0034] In certain embodiments, the compound of formula I and / or solvates and / or hydrates thereof or the composition or the medicament is administered to a subject by injection in a dose of 1 mg / kg to 1000 mg / kg body weight (active ingredient).

[0035] In certain embodiments, the Bunyavirus is a Nairovirus.

[0036] In certain embodiments, the Bunyavirus is a Orthonairovirus.

[0037] In certain embodiments, the Bunyavirus is Crimean-Congo hemorrhagic fever virus (CCHFV).

[0038] In certain embodiments, the disease caused by the Bunyavirus is a simple infection, fever, headache, muscle pain, vomiting, gastrointestinal bleeding, nosebleed, hematemesis or shock caused by a Bunyavirus.

[0039] In certain embodiments, the disease caused by the Bunyavirus is hemorrhagic fever caused by a Nairovirus.

[0040] In certain embodiments, the disease caused by the Bunyavirus is hemorrhagic fever caused by a Orthonairovirus.

[0041] In certain embodiments, the disease caused by the Bunyavirus is hemorrhagic fever caused by Crimean-Congo hemorrhagic fever virus.

[0042] In certain embodiments, the disease caused by the Bunyavirus is Crimean-Congo hemorrhagic fever.

[0043] In certain embodiments, the mammal includes a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, cat, dog, or pig.

[0044] In certain embodiments, the subject includes a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, cat, dog, or pig.

[0045] In certain embodiments, the mammal includes a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, cat, dog, or pig.

[0046] It is further noted that the specific dosage and method of administration of the compound of Formula I and / or solvate and / or hydrate thereof will be decided by the attending physician within the scope of sound medical judgment. The dose employed will depend on various factors, including the age, body weight, sex, natural health status, nutritional status, drug activity, time of administration, metabolic rate, severity of the disease condition being treated, and the judgment of the medical practitioner. Preferably, the dosage employed is between 0.001 and 1000 mg / kg body weight / day, such as 1 mg / kg to 1000 mg / kg body weight / day.

[0047] The compositions described herein can be prepared in various forms for different routes of administration, such as injections, including powder injections and injection solutions. BRIEF DESCRIPTION OF DRAWINGS

[0048] Figure 1 is the results of in vitro anti-Crimean-Congo hemorrhagic fever virus experiment of baloxavir sodium of the present application;

[0049] Figure 2 is the results of pharmacodynamics experiment of baloxavir sodium of the present application in mice. DETAILED DESCRIPTION

[0050] The following examples are illustrative preferred embodiments of the application and do not pose any limitation on the application.

[0051] Example 1: Preparation of Sodium Baloxavir

[0052] Baloxavir acid 4.9 g (0.01 mol) was added to 20 mL of water, 1 N NaOH 20 mL (0.02 mol) was added, and the reaction was carried out at 80 °C for 12 h. The reaction was stopped, and white solid 4.5 g was obtained by suction filtration, with a yield of 88.8%.

[0053] Example 2: Study on the solubility of sodium baloxavir

[0054] According to the Chinese Pharmacopoeia (2020 edition), the equilibrium solubility in pure water was determined by the room temperature shake flask method. The purity of sodium baloxavir used in the study was 99.5%. The powdered sample was weighed and placed in a certain volume of solvent, and shaken thoroughly for 30 seconds every 5 minutes at room temperature. The dissolution behavior of the sample was observed for 30 minutes. If no solute particles were observed, it was considered that the solute had completely dissolved. At the same time, a control example was set up to compare the dissolution performance of sodium baloxavir, marbovir and baloxavir.

[0055] As shown in Table 1, sodium baloxavir has high water solubility.

[0056] Table 1 Compound solubility test

[0057] Example 3: In vitro screening experiment of sodium baloxavir against Crimean-Congo hemorrhagic fever virus

[0058] 3.1 Drug treatment of virus-infected cells

[0059] Human umbilical vein endothelial cells (HUVEC) were cultured in Eagle's medium (Gibco, Rockville, MD, USA) at 37 °C with a minimum of 5% CO2, with the addition of 10% fetal bovine serum (Gibco). The CCHFV strain IbAr10200-eGFP used in the study was preserved in the National Virus Resource Center (Wuhan, China). All CCHFV infection experiments were carried out in a BLS-3 laboratory.

[0060] 3.2 Cell toxicity determination

[0061] Well-conditioned HUVEC cells were seeded at about 1 × 10 4Each well was plated with 10 μL of CCK-8 solution (Beyotime, China) and the culture plate was placed in an incubator for 1-4 hours. The absorbance at 450 nm was measured with an enzyme-labeled instrument to detect the median cytotoxic concentration (CC) of the compound. 50 ).

[0062] 3.3 Viral load detection

[0063] To evaluate the activity of antiviral drugs, HUVEC cells (approximately 6 × 10 4 / well) were pre-seeded on a 48-well plate, serially diluted drugs were added, and the cells were infected after incubation at 37°C for 1 hour. After infecting the cells at 37°C with a multiplicity of infection (MOI) of 0.01 for 1 hour, the cell supernatant was discarded and the cells were cultured with fresh drug-containing culture medium. 72 hours after infection (pi), 100 μL of supernatant was collected for RNA extraction, and the cells were subjected to IFA or western detection respectively. Then, the RNA copy number of the supernatant was quantified by real-time quantitative polymerase chain reaction (qRT-PCR), and the median effective concentration (EC) was further calculated. 50 The primers for amplifying CCHFV S segment were: (qF) 5′-TCAAGTGGAGGAAGGACATAGG-3′ and (qR) 5′-TCCACATGTTCACGGCTCACTGGG-3′.

[0064] Total RNA was extracted using TRIzol reagent (Takara) according to the manufacturer's instructions, and qRT-PCR was performed using 1 μg of total RNA as described above.

[0065] 3.4 Experimental Results

[0066] To preliminarily evaluate the in vitro anti-CCHFV activity of baloxavir sodium, a screening assay was performed using two drug concentrations (20 μM and 5 μM) and the positive control T-705. HUVEC cells were infected in the presence of either drug or DMSO, and fluorescence images were taken every 48 hours to quantify the infection rate. At concentrations of 20 μM and 5 μM, baloxavir sodium inhibited CCHFV infection by nearly 100% and 90%, respectively. In contrast, at the same concentrations, T-705 only inhibited viral infection by approximately 80% and 30%.

[0067] Furthermore, the EC values ​​of baloxavir sodium and T-705 were quantitatively determined by qRT-PCR. 50 As shown in Figure 1, the EC of baloxavir 50 The value was lower, 0.6 μM, while the EC 50 The 50% cytotoxic concentration (CC) of baloxavir sodium and T-705 was 4.2 μM. 50 ) were 83.6 μM and >100 μM, respectively, and the selectivity index (SI = CC 50 / EC 50 ) were 139.3 and >23.8, respectively. These results indicate that, under this infection condition, baloxavir sodium has better anti-CCHFV activity in vitro than T-705.

[0068] Example 4: Pharmacokinetic Study of Baloxavir Sodium in Rats

[0069] The experiment used SD rats (male) and divided them into an oral administration group or an intraperitoneal injection administration group. Baloxavir sodium and Tween 80 were prepared into an injection solution and then administered accordingly. Blood samples were collected at different time points after administration (5min, 15min, 30min, 1h, 2h, 4h, 8h, 12h, 12.083h, 12.25h, 12.5h, 13h, 14h, 24h). Within 2h, the whole blood was centrifuged at 2500G for 10min at 4℃, separated and placed in EP tubes, and stored in a -20℃ refrigerator. For sample analysis, 10 μl of plasma was added to 10 μl of blank plasma and vortexed to mix. 20 μl of acetonitrile and 100 μl of acetonitrile precipitant (containing 5 ng / ml propranolol acetonitrile solution) were added. The sample was vortexed for 1 min and centrifuged at 14000 rpm for 10 min at 4°C. 100 μl of the supernatant after centrifugation was added to 400 μl of diluent (50% acetonitrile aqueous solution), and the sample was analyzed using LC-MS / MS8050 after thorough vortexing.

[0070] The pharmacokinetic characteristics of baloxavir sodium were observed in SD rats after administration. As shown in Table 2, compared with oral administration of baloxavir sodium (AUC 0-t =0.544±0.269μg·h / mL; Cmax = 0.062 ± 0.022 pg / mL) compared to 75 mg / kg i.p. dose, the plasma exposure of baloxavir sodium was AUC 0-t = 23.49 ± 6.08 pg-h / mL and maximum plasma concentration C max = 1.82 ± 0.5 pg / mL. The above results indicated that i.p. administration of baloxavir sodium had good pharmacokinetic properties in rats.

[0071] Table 2 Pharmacokinetic test of compound baloxavir sodium

[0072] Example 5: Experiment of baloxavir sodium injection protecting mice from death infected with Crimean-Congo hemorrhagic fever virus CCHFV

[0073] All animal experiments used type I IFN receptor knockout (IFNAR - / - ) mice on C57BL / 6J background. 7-12 weeks old female mice were infected with CCHFV-YL16070 strain (3000 TCID 50 ) by intraperitoneal injection (i.p.). The mice were given baloxavir sodium by i.p. administration, and the control group was given similar solvent injection. The body weight and clinical symptoms of the mice were monitored every day. When the mice lost more than 20% of their body weight, did not respond to touch stimulation, and had difficulty climbing, they were euthanized with isoflurane anesthetic. Statistical analysis was performed using two-way analysis of variance (ANOVA) test. The livers of the mice were collected for detection of viral load and pathological analysis. The survival percentage P value was determined by log-rank test. Statistical analysis was performed using unpaired t-test.

[0074] We evaluated the in vivo efficacy of baloxavir sodium against CCHFV. Baloxavir sodium Tween 80 injection was administered at a dose of 75 mg / kg twice daily by i.p. for 3 days. The treatment group showed a protective effect on mice infected with CCHFV. The study showed that the total survival rate of the 75 mg / kg twice daily administration regimen was 83.3% (Figure 2A) (n = 6 per group), while the survival rate of the blank control group was 16.7%. After treatment, mice given a dose of 75 mg / kg twice daily had a liver viral load that was reduced by about 2 log copies / mg tissue on day 3 (*P < 0.05) and day 6 (*P < 0.01) (Figures 2C, D). The weight loss phenomenon of mice treated with baloxavir sodium was also alleviated compared to the control group (Figure 2B).

[0075] It should be noted that the above examples are only used to illustrate the technical solutions of the present application but not to limit the same; although the present application has been described in detail with reference to the preferred embodiments, it is understood by those skilled in the art that the specific implementation of the present application can be modified or some technical features can be replaced by equivalent ones; without departing from the spirit of the technical solutions of the present application, all of them should be covered in the technical solution range claimed by the present application.

Claims

1. Use of a compound of formula I and / or its solvate and / or its hydrate in the preparation of a medicament for treating a disease or infection caused by a bunyavirus in a subject, Preferably, the medicament is administered to the subject by injection. Preferably, the medicament is administered to the subject by intravenous, intramuscular or other injection. Preferably, the medicament is administered to the subject by injection at a dose of 1 mg / kg to 1000 mg / kg body weight (based on the active ingredient).

2. Use of a composition in the manufacture of a medicament for treating a disease or infection caused by a Bunyavirus in a subject, wherein the composition comprises a compound of Formula I and / or solvate thereof and / or hydrate thereof, Preferably, the medicament is administered to the subject by injection. Preferably, the medicament is administered to the subject by intravenous, intramuscular or other injection. Preferably, the medicament is administered to the subject by injection at a dose of 1 mg / kg to 1000 mg / kg body weight (based on the active ingredient). Preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the composition is an injection. Preferably, the injection comprises: (1) a compound of Formula I and / or solvate and / or hydrate thereof, in an amount of 0.1-20%; (2) a solvent selected from the group consisting of water for injection, physiological saline, sterile water and the like, in an amount of 60-99%; (3) one or more excipients selected from the group consisting of: a pH adjusting agent such as hydrochloric acid, acetic acid and the like, in an amount of 0.01-5%; an isotonic agent such as mannitol, sodium chloride and the like, in an amount of 0.1-10%; a preservative such as p-hydroxybenzoic acid ester and the like, in an amount of 0.01-1%; a stabilizer such as ethanol, propylene glycol and the like, in an amount of 0.1-10%; Preferably, the pH of the injection is 4-9.

3. An injection comprising a compound of Formula I and / or solvate and / or hydrate thereof, Preferably, the injection comprises: (1) a compound of Formula I and / or solvate and / or hydrate thereof, in an amount of 0.1-20%; (2) a solvent selected from the group consisting of water for injection, physiological saline, sterile water and the like, in an amount of 60-99%; (3) one or more excipients selected from the group consisting of: a pH adjusting agent such as hydrochloric acid, acetic acid and the like, in an amount of 0.01-5%; an isotonic agent such as mannitol, sodium chloride and the like, in an amount of 0.1-10%; a preservative such as p-hydroxybenzoic acid ester and the like, in an amount of 0.01-1%; a stabilizer such as ethanol, propylene glycol and the like, in an amount of 0.1-10%; Preferably, the pH of the injection is 4-9.

4. Use of a composition for the manufacture of a medicament as a Bunyavirus inhibitor, wherein the composition comprises a compound of Formula I and / or solvate thereof and / or hydrate thereof, Preferably, the medicament is administered to the subject by injection. Preferably, the medicament is administered to the subject by intravenous, intramuscular or other injection. Preferably, the medicament is administered to the subject by injection at a dose of 1 mg / kg to 1000 mg / kg body weight (based on the active ingredient). Preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the composition is an injection. Preferably, the injection comprises: (1) a compound of Formula I and / or solvate and / or hydrate thereof, in an amount of 0.1-20%; (2) a solvent selected from the group consisting of water for injection, physiological saline, sterile water and the like, in an amount of 60-99%; (3) one or more excipients selected from the group consisting of: a pH adjusting agent such as hydrochloric acid, acetic acid and the like, in an amount of 0.01-5%; an isotonic agent such as mannitol, sodium chloride and the like, in an amount of 0.1-10%; a preservative such as p-hydroxybenzoic acid ester and the like, in an amount of 0.01-1%; an isotonic agent, such as mannitol, sodium chloride, etc., in an amount of 0.1-10%; a preservative, such as p-hydroxybenzoate, etc., in an amount of 0.01-1%; a stabilizer, such as ethanol, propylene glycol, etc., in an amount of 0.1-10%; Preferably, the pH of the injection is 4-9.

5. Use of a composition for the manufacture of a medicament for inhibiting replication or propagation of a Bunyavirus in a cell (e.g. a mammalian cell) in or ex vivo, wherein the composition comprises a compound of Formula I and / or solvates thereof and / or hydrates thereof, Preferably, the drug is administered to the subject by injection. Preferably, the drug is administered to the subject by intravenous, intramuscular or other injection. Preferably, the drug is administered to the subject by injection at a dosage of 1 mg / kg to 1000 mg / kg body weight (active ingredient). Preferably, the composition further comprises a pharmaceutically acceptable carrier or excipient. Preferably, the composition is an injection. Preferably, the injection comprises: (1) a compound of Formula I and / or solvate and / or hydrate thereof in an amount of 0.1-20%; (2) a solvent selected from water for injection, physiological saline, sterile water, etc. in an amount of 60-99%; (3) one or more excipients selected from: a pH adjusting agent, such as hydrochloric acid, acetic acid, etc., in an amount of 0.01-5%; an isotonic agent, such as mannitol, sodium chloride, etc., in an amount of 0.1-10%; a preservative, such as p-hydroxybenzoate, etc., in an amount of 0.01-1%; a stabilizer, such as ethanol, propylene glycol, etc., in an amount of 0.1-10%; Preferably, the pH of the injection is 4-9.

6. Use of a compound of formula I ###0002### and / or solvates thereof and / or hydrates thereof for the manufacture of a medicament as inhibitor of a Bunyavirus, Preferably, the drug is administered to the subject by injection. Preferably, the drug is administered to the subject by intravenous, intramuscular or other injection. Preferably, the drug is administered to the subject by injection at a dosage of 1 mg / kg to 1000 mg / kg body weight (active ingredient).

7. Use of a compound of Formula I and / or solvate thereof and / or hydrate thereof in the manufacture of a medicament for inhibiting replication or propagation of a Bunyavirus in a cell (e.g., a mammalian cell) in vivo or in vitro, Preferably, the drug is administered to the subject by injection. Preferably, the drug is administered to the subject by intravenous, intramuscular or other injection. Preferably, the drug is administered to the subject by injection at a dosage of 1 mg / kg to 1000 mg / kg body weight (active ingredient).

8. The use according to any one of claims 1-7, wherein the Bunyavirus is a Nairovirus, Preferably, the Bunyavirus is a Orthonairovirus, Preferably, the Bunyavirus is Crimean-Congo hemorrhagic fever virus (CCHFV).

9. The use according to any one of claims 1-7, wherein the disease caused by the Bunyavirus is a simple infection, fever, headache, muscle pain, vomiting, gastrointestinal bleeding, nosebleed, hematemesis or shock caused by the Bunyavirus; Preferably, the disease caused by the Bunyavirus is hemorrhagic fever caused by the Nairovirus; Preferably, the disease caused by the Bunyavirus is hemorrhagic fever caused by the Orthonairovirus; Preferably, the disease caused by the Bunyavirus is hemorrhagic fever caused by the Crimean-Congo hemorrhagic fever virus.

10. The use according to claims 1-3, wherein the disease caused by the Bunyavirus is Crimean-Congo hemorrhagic fever.

11. The use of any one of claims 1-7, wherein the mammal or the subject comprises a bovine, equine, ovine, porcine, canine, feline, rodent, primate, such as a human, cat, dog, or pig.

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