A compound for stabilizing virus-based therapeutic agents.

Abstract

This invention relates to a viral preparation for administration to a subject.

Description

[Technical Field]

[0001] Field of Invention The present invention relates to pharmaceutical formulations for virus-based therapeutic agents and virus-based cancer vaccines. More specifically, the present invention relates to pharmaceutical formulations for oncolytic viruses as disclosed herein.

[0002] Background of the Invention Oncolytic viruses, such as vesicular stomatitis virus (VSV) (VSV-GP) containing the glycoprotein (GP) of lymphocytic choriomeningitis virus, represent an emerging class of biopharmaceuticals that selectively replicate in cancer cells and selectively kill them. The oncolytic viruses described herein can spread within tumors and efficiently induce lysis of tumor cells, resulting in cell death. Furthermore, additional genes may be cloned into the viral genome, and the expression of such proteins can stimulate the immune system and / or direct the immune system towards tumor cells. Moreover, oncolytic viruses expressing cancer-specific antigens may be co-administered together with the antigens, thereby enhancing and prolonging their immunostimulatory effects. Therefore, oncolytic viruses are beneficial for the treatment and / or prevention of cancer.

[0003] Developing formulations to stabilize oncolytic viruses during storage is challenging but rewarding. Isolated and concentrated live viruses are typically unstable during long-term storage and are sensitive to high temperatures, mechanical stress, and freeze-thaw cycles. To be effective as a therapeutic agent, live viruses need to be formulated so that their activity is preserved and agglutination is prevented, thus preventing the formation of visible and invisible particles (SvPs). Viruses, particularly VSV-GP, tend to self-associate and agglutinate due to their structural complexity. Colloidal stability of the virus particles must be ensured, and agglutination must be reduced to below the limits specified in the official standards through formulation development efforts. For example, the number of freeze-thaw events for filling and labeling vials, as well as the freezing and storage conditions, such as the rate of freezing and thawing and the storage temperature, should be considered so that the virus does not reach the glass transition temperature (T) of the maximum freeze-concentrated solution. g Infectivity and colloidal stability are significantly affected, especially if they are not stored as a frozen liquid at temperatures below ').

[0004] While several formulations for virus-based therapeutics for specific viruses are described in the literature, it is generally understood that each virus species, genus, or family needs to be formulated with its own specific prescription.

[0005] Therefore, there is a need for stable liquid, frozen liquid, and dried formulations (such as lyophilized formulations) for virus-based therapeutic agents suitable for parenteral administration, including intravenous or intratumoral injection into human patients, particularly formulations for oncolytic viruses, such as VSV, especially VSV-GP. There is also a need for formulations that exhibit enhanced colloidal stability and show no or only minimal loss of the biological activity of the therapeutic virus during long-term storage at various temperatures.

[0006] Summary of the Invention The present invention addresses the above-mentioned need by providing stable formulations for virus-based therapeutic agents, particularly stable liquid and frozen liquid formulations.

[0007] In a first embodiment, the present invention relates to a pharmaceutical composition comprising an enveloped virus and a protein substance and / or a poly(ethylene oxide) / poly(propylene oxide) block copolymer.

[0008] In one embodiment relating to the first aspect, the poly(ethylene oxide) and poly(propylene oxide) block copolymer is a poloxamer. In one embodiment relating to the first aspect, the poly(ethylene oxide) and poly(propylene oxide) block copolymer is a poloxamer 188.

[0009] In one embodiment relating to the first aspect, the protein substance is albumin or gelatin. In one embodiment relating to the first aspect, the protein substance is human serum albumin or recombinant human albumin.

[0010] In one embodiment relating to the first aspect, the poly(ethylene oxide) and poly(propylene oxide) block copolymer is poloxamer 188, and the protein substance is human serum albumin or recombinant human albumin. In the related embodiments, the pharmaceutical composition is 0.01-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.01-40 g / L, 0.01-30 g / L, 0.01-20 g / L L, 0.01~10g / L, 0.01~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0.1~10g / L, 0.1~5g / L, 0.2 ~40g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0.3~30g / L, 0.3~20g / L, 0.3~10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, 1~10g / L, 1~ Contains poloxamer 188 at concentrations of 5 g / L, 2-40 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L.In further related embodiments, the pharmaceutical composition is expressed in concentrations of 0.05-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.05-40 g / L, 0.05-30 g / L, and 0.05-20 g / L. L, 0.05~10g / L, 0.05~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0.1~10g / L, 0.1~5g / L, 0.2~4 0g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0.3~30g / L, 0.3~20g / L, 0.3~ 10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~40g / L, 0.5 ~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, 1~10g / L, 1~5g / L, 2~4 Contains human serum albumin or recombinant human albumin at concentrations of 0 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L.

[0011] In one embodiment related to the first aspect and in any of the group of embodiments thereof, the pharmaceutical composition further comprises at least one of an amino acid, a buffer, or a sugar. In the related embodiment, the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, and is preferably arginine. In the related embodiment, the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, and is preferably Tris. In the related embodiment, the buffer has a concentration of 1-100 mM, 1-90 mM, 1-80 mM, 1-70 mM, 1-60 mM, 1-50 mM, 1-40 mM, 1-30 mM, 1-20 mM, or 1-10 mM. In related embodiments, the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, and is preferably trehalose. In related embodiments, the sugar is expressed in concentrations of 10-1000 mM, 10-900 mM, 10-800 mM, 10-700 mM, 10-600 mM, 10-500 mM, 10-400 mM, 10-300 mM, 10-200 mM, 20-1000 mM, 20-900 mM, 20-800 mM, 20-700 mM, 20-600 mM, 20-500 mM, 20-400 mM, 20-300 mM, 20-200 mM, 30-1000 mM, 30-900 mM, 30-800 mM, 30-700 mM, and 3 It has concentrations of 0-600 mM, 30-500 mM, 30-400 mM, 30-300 mM, 30-200 mM, 40-1000 mM, 40-900 mM, 40-800 mM, 40-700 mM, 40-600 mM, 40-500 mM, 40-400 mM, 40-300 mM, 40-200 mM, 50-1000 mM, 50-900 mM, 50-800 mM, 50-700 mM, 50-600 mM, 50-500 mM, 50-400 mM, 50-300 mM, or 50-200 mM.

[0012] In one embodiment related to the first aspect and in any of the group of embodiments thereof, the composition further comprises one or more sugar alcohols. In the related embodiment, one or more sugar alcohols are selected from the group consisting of mannitol, sorbitol, xylitol, maltitol, maltitol syrup, lactitol, inositol, glycerol, erythritol, isomalt, and hydrolyzed hydrogenated starch. In a further related embodiment, one or more sugar alcohols are mannitol and / or sorbitol, preferably a combination of mannitol and sorbitol.

[0013] In one embodiment relating to the first aspect, the composition is substantially chloride-free, preferably substantially sodium chloride-free.

[0014] In one embodiment relating to the first aspect, the pH of the composition is 5 to 9, or 6 to 9, or 6.5 to 8.5, or 6.5 to 8.0, preferably 7.0 to 8.0. In the related embodiment, the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

[0015] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - A buffer solution, at least one of an amino acid or a sugar, and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0016] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffer solutions, and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0017] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and sugars, and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0018] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids, buffer solutions, and sugars, - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0019] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer and sugar, and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0020] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - A buffer, at least one of amino acids or sugars (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate and Tris, preferably Tris), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0021] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffer (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0022] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids, buffers, and sugars (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0023] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer and sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0024] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - A buffer solution, at least one of amino acids or sugars (wherein sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0025] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffer solutions (wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0026] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and sugars (wherein sugars are selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0027] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffers and sugars (wherein sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0028] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer solution and sugar (wherein sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0029] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - A buffer solution, at least one of an amino acid or a sugar (wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0030] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and sugars (wherein amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0031] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffer solutions and sugars (wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0032] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - A buffer, an amino acid, or at least one of the following (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0033] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and buffers (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0034] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids and sugars (wherein amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein sugars are selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0035] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids, buffer solutions, and sugars (wherein the buffer solution is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0036] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer and sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine and glutamine, preferably arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0037] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0038] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids (where the amino acid is arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0039] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Sugars (where sugar is trehalose or sucrose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0040] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Amino acids (in this case, the amino acid is arginine), - Sugars (where sugar is trehalose or sucrose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0041] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Sugars (where sugar is trehalose or sucrose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0042] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Amino acids (where the amino acid is arginine), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0043] In one embodiment relating to the first aspect, the pharmaceutical composition is - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Amino acids (in this case, the amino acid is arginine), - Sugars (where sugar is trehalose or sucrose), and - Protein materials and / or poly(ethylene oxide) / poly(propylene oxide) block copolymers (The protein substance here is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.) Includes.

[0044] In one embodiment and any of the embodiments relating to the first embodiment, which includes a buffer, the buffer has a concentration of 1-100 mM, 1-90 mM, 1-80 mM, 1-70 mM, 1-60 mM, 1-50 mM, 1-40 mM, 1-30 mM, 1-20 mM, or 1-10 mM.

[0045] In one embodiment and any of the embodiments relating to the first embodiment, which contains sugar, the sugar is present in concentrations of 10-1000 mM, 10-900 mM, 10-800 mM, 10-700 mM, 10-600 mM, 10-500 mM, 10-400 mM, 10-300 mM, 10-200 mM, 20-1000 mM, 20-900 mM, 20-800 mM, 20-700 mM, 20-600 mM, 20-500 mM, 20-400 mM, 20-300 mM, 20-200 mM, 30-1000 mM, 30-900 mM, and 30 It has concentrations of ~800mM, 30~700mM, 30~600mM, 30~500mM, 30~400mM, 30~300mM, 30~200mM, 40~1000mM, 40~900mM, 40~800mM, 40~700mM, 40~600mM, 40~500mM, 40~400mM, 40~300mM, 40~200mM, 50~1000mM, 50~900mM, 50~800mM, 50~700mM, 50~600mM, 50~500mM, 50~400mM, 50~300mM, or 50~200mM.

[0046] In one embodiment related to the first aspect and any of the group of embodiments thereof, the pharmaceutical composition is 0.01-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.01-40 g / L, 0 .01~30g / L, 0.01~20g / L, 0.01~10g / L, 0.01~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0.1~10 g / L, 0.1~5g / L, 0.2~40g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0.3~30g / L, 0.3~20g / L, 0.3~10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, 1~1 Contains poloxamer 188 at concentrations of 0 g / L, 1-5 g / L, 2-40 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L.

[0047] In one embodiment related to the first aspect and any of the group of embodiments thereof, the pharmaceutical composition is 0.05-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.05-40 g / L, 0.05 ~30g / L, 0.05~20g / L, 0.05~10g / L, 0.05~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0.1~10g / L, 0 .1~5g / L, 0.2~40g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0.3~30g / L, 0.3 ~20g / L, 0.3~10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~4 0g / L, 0.5~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, 1~10g / L, 1~5g Contains human serum albumin or recombinant human albumin at concentrations of 1 / L, 2-40 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L.

[0048] In one embodiment relating to the first aspect, the pharmaceutical composition comprises about 1 to 100 mM tris, about 10 to 500 mM arginine, about 10 to 500 mM trehalose, about 0.1 to 5 mg / ml poloxamer 188, about 0.5 to 10 mg / ml recombinant human albumin, and a pH of about 6 to 8.

[0049] In one embodiment relating to the first aspect, the pharmaceutical composition comprises an enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 100 mM Trehalose, about 0.5 mg / ml Poloxamer 188, about 2 mg / ml Recombinant Human Albumin, and a pH of about 7.5.

[0050] In one embodiment related to the first aspect and any of its groups of embodiments, the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

[0051] In one embodiment related to the first aspect and any of its groups of embodiments, the enveloped virus is a rhabdovirus, preferably vesiculovirus or vesicular stomatitis virus (VSV). In related embodiments, the enveloped virus is a recombinant vesicular stomatitis virus (VSV), where the gene encoding the glycoprotein G of the vesicular stomatitis virus here is replaced by the gene encoding the glycoprotein GP of lymphocytic choriomeningitis virus (LCMV), and / or the glycoprotein G is replaced by the glycoprotein GP of LCMV. In related embodiments, the pharmaceutical composition contains at least 1×10 5 of TCID 50 / mL, at least 1×10 6 of TCID 50 / mL, at least 1×10 7 of TCID 50 / mL, at least 1×10 8 of TCID 50 / mL, at least 1×10 9 of TCID 50 / mL, or at least 1×10 9 of TCID 50 / mL concentration of an enveloped virus, preferably vesiculovirus or vesicular stomatitis virus (VSV). In further related embodiments, the pharmaceutical composition contains 1×10 5 of TCID 50 / mL to 1×10 12 of TCID 50 / mL, 1×10 6 of TCID 50 / mL to 1×10 12 of TCID 50 / mL, 1×10 7 of TCID 50 / mL to 1×10 12 of TCID 50 / mL, 1×108 TCID 50 / mL ~ 1 × 10 12 TCID 50 / mL, 1 × 10 5 TCID 50 / mL ~ 1 × 10 11 TCID 50 / mL, 1 × 10 5 TCID 50 / mL ~ 1 × 10 10 TCID 50 / mL, or 1 × 10 5 TCID 50 / mL ~ 1 × 10 9 TCID 50 It contains an enveloped virus, preferably becyclovirus or vesicular stomatitis virus (VSV), in a concentration range of / mL.

[0052] In one embodiment related to the first aspect and in any of the group of embodiments thereof, the pharmaceutical composition is a liquid or frozen liquid pharmaceutical composition. In the related embodiments, the composition is stored frozen at temperatures of about -80°C, -70°C, -60°C, -50°C, -40°C, -35°C, -30°C, -25°C, -20°C, -15°C, -10°C, or -5°C. In the related embodiments, the present invention relates to a product produced by freezing a liquid pharmaceutical composition. Further embodiments relate to a pharmaceutical composition comprising water and the product.

[0053] In one embodiment relating to the first aspect and in any of the group of embodiments thereof, the present invention relates to a dry pharmaceutical composition produced by a method comprising the step of removing water from a pharmaceutical composition described in the first aspect and in any of the group of embodiments thereof. In the related embodiment, a pharmaceutical composition containing ice is obtained by freezing the dry pharmaceutical composition, and then the water is removed. In the related embodiment, the method further comprises the step of placing the liquid pharmaceutical composition in a vacuum under controlled temperature and pressure to remove water. In a further related embodiment, the method is freeze-drying. In a further related embodiment, the dry pharmaceutical composition contains less than about (0.5% to 5%) w / w of water. Further embodiments relate to pharmaceutical compositions containing water and dry pharmaceutical compositions. [Brief explanation of the drawing]

[0054] [Figure 1] Geometric mean infectivity of VSV-GP-Cargo 1 after storage of liquid formulations at 5°C. Error bars indicate standard deviation (n=2 for control, n=3 for others, measured with 3 replicates each). Frozen crude samples serve as controls. [Figure 2] Geometric mean infectivity of VSV-GP-Cargo 1 after storage of liquid formulations at 25°C. Error bars indicate standard deviation (n=2 for control, n=3 for others, measured with 3 replicates each). Frozen crude samples serve as controls. [Figure 3] Geometric mean values ​​of SvPs (invisible particles) 10 μm or larger in VSV-GP-Cargo 1 after storing the liquid formulation at 5°C. Error bars indicate the standard deviation (n=2 for the control, n=3 for the others, measured with three replicates each). [Figure 4] Geometric mean of SvPs ≥ 10 μm in VSV-GP-Cargo 1 after storing liquid formulations at 25°C. Error bars indicate standard deviation (n=2 for control, n=3 for others, measured with 3 replicates each). [Figure 5]Geometric mean infectivity of VSV-GP-Cargo 1 after 0, 1, or 3 freeze / thaw cycles and storage of liquid formulations at 25°C. Error bars indicate standard deviation (n=2 for controls, n=3 for others, measured with 3 replicates each). Frozen crude samples serve as controls. [Figure 6] Geometric mean of SvPs ≥ 10 μm in VSV-GP-Cargo 1 after 0, 1, or 3 freeze / thaw cycles of the liquid formulation. Error bars indicate the standard deviation (n=2 for the control, n=3 for the others, measured with 3 replicates each). [Figure 7] Geometric mean infectivity of VSV-GP-Cargo 1 after one, three, or five freeze / thaw cycles at -20°C and +25°C for frozen liquid formulations. Error bars indicate standard deviation (n=2 for controls, n=3 for others, measured with three replicates each). Frozen crude samples serve as controls. [Figure 8] Geometric mean infectivity of VSV-GP-Cargo 1 after one, three, or five freeze / thaw cycles at -80°C and +25°C for frozen liquid formulations. Error bars indicate standard deviation (n=2 for controls, n=3 for others, measured with three replicates each). Frozen crude samples serve as controls. [Figure 9] Geometric mean of SvPs ≥ 10 μm in VSV-GP-Cargo 1 after one, three, or five freeze / thaw cycles at -20°C and +25°C for frozen liquid formulations. Error bars indicate standard deviation (n=2 for control, n=3 for others, measured with three replicates each). [Figure 10] Geometric mean of SvPs ≥ 10 μm in VSV-GP-Cargo 1 after one, three, or five freeze / thaw cycles at -80°C and +25°C for frozen liquid formulations. Error bars indicate standard deviation (n=2 for control, n=3 for others, measured with three replicates each). [Figure 11]Geometric mean of infectivity of VSV-GP-Cargo 1 after storage of freeze-dried preparations at 25°C. Error bars indicate standard deviation (n=2 for controls, n=3 for others, measured with 3 replicates each). Freeze-dried crude samples serve as controls. The first time point shows the measurement before freeze-drying. The second value (week 0) is the value immediately after freeze-drying and reconstitution. [Figure 12] Geometric mean values ​​of SvPs ≥ 10 μm in VSV-GP-Cargo 1 after storage of freeze-dried formulations at 25°C. Error bars indicate the standard deviation (n=2 for the control, n=3 for the others, measured with three replicates each). The first time point shows the measurement before freeze-drying. The second value is the value immediately after freeze-drying and reconstitution. [Figure 13] Geometric mean values ​​of SvPs ≥ 2 μm in VSV-GP after storing the frozen liquid formulation at 25°C for 1 week (T1w) or 2 weeks (T2w), or after one freeze / thaw cycle (T-FT) (measured with n=1 and 3 replicates). Raw data were converted considering a 100-fold dilution. The initial t0 value is the value before freezing. [Figure 14] Geometric mean values ​​of SvPs ≥ 10 μm in VSV-GP after storing the frozen liquid formulation at 25°C for 1 week (T1w) or 2 weeks (T2w), or after one freeze / thaw cycle (T-FT) (each measured with n=1 and three replicates). Raw data were converted considering a 100-fold dilution. The initial t0 value is the value before freezing. [Figure 15] Geometric mean (n=1) of the infectivity of VSV-GP after storing the frozen liquid preparation at 25°C for one week (T1w) or two weeks (T2w), or after one freeze / thaw cycle (T-FT). The initial t0 value is the value before freezing. [Figure 16] An overview of the infectivity titers of VSV-GP DP (dried preparation) samples at various time points, determined by TCID50 (50% tissue culture infectious dose) analysis (n=3), after lyophilization (T-FT) or storage at 25°C for 1 week (T-1w) or 2 weeks (T-2w). The starting material was VSV-GP in elution buffer. [Figure 17]Geometric mean of SvPs of VSV-GP 2 μm or larger after storing the frozen liquid formulation at 25°C for one week (T1w) or two weeks (T2w), or after one freeze / thaw cycle (T-FT) (n=1). Raw data was multiplied by a dilution factor of 20. The initial t0 value is the value before freezing. [Figure 18] Geometric mean of SvPs 10 μm or larger in the VSV-GP dry preparation after storing the frozen liquid preparation at 25°C for one week (T1w) or two weeks (T2w), or after one freeze / thaw cycle (T-FT) (n=1). Raw data was converted considering a 20-fold dilution. The initial t0 value is the value before freezing. [Figure 19] Geometric mean of VSV-GP infectivity after storing the frozen liquid preparation at 25°C for one week (T1w) or two weeks (T2w), or after one freeze / thaw cycle (T-FT). Error bars indicate the standard deviation (each measured with n=4 and three replicates). The initial t0 value is the value before freezing. VSV-GP batches with a titer of 1–2 × 10⁹ TCID50 / ml serve as controls. [Figure 20] Geometric mean of infectivity of VSV-GP after freeze-thaw (T-FT) or after storage at 25°C for two weeks (T-2w 25°C). Error bars indicate the standard deviation (measured with n=4 and 3 replicates).

[0055] Detailed description of the invention In the following detailed description, specific numerical details are provided to give a full understanding of the invention. However, it will be apparent to those skilled in the art that the subject art can be practiced without using some of these specific details. In other cases, well-known structures and techniques are not described in detail in order not to make the invention difficult to understand. Headings are included simply for convenience to aid in interpretation and should not be understood as limiting the invention to any particular aspect or embodiment.

[0056] In one embodiment, the formulation of the present invention is useful for stabilizing the infectivity titer of enveloped viruses. In another embodiment, the formulation helps maintain the infectivity titer and / or colloidal stability of enveloped viruses after one or more freeze-thaw cycles. In a related embodiment, the formulation is useful for maintaining the infectivity titer and / or colloidal stability of enveloped viruses during storage at room temperature, or even above room temperature, at elevated temperatures, preferably 2 to 8°C. In a related embodiment, the formulation is useful for maintaining the titer and / or activity of enveloped viruses during storage of the virus at various temperatures over a period of time.

[0057] In another embodiment, the preparations of the present invention can reduce and / or slow down the formation of visible and / or invisible particles (SvPs) in preparations containing enveloped viruses. In a related embodiment, the preparations containing enveloped viruses have reduced amounts of visible viruses and / or invisible particles.

[0058] In yet another embodiment, the formulation of the present invention helps maintain the infectivity titer of enveloped viruses. In a related embodiment, the infectivity titer is maintained or the decline in the infectivity titer over time is slowed. In a related embodiment, the decline in the infectivity titer due to long-term storage, repeated freeze-thaw cycles, storage at elevated temperatures, or mechanical stress is slowed.

[0059] In another embodiment, the preparations of the present invention help to inhibit, slow down, or prevent the aggregation of enveloped viruses. In another embodiment, the preparations are useful for inhibiting, reducing, or preventing turbidity in preparations containing enveloped viruses.

[0060] In another embodiment, the preparations of the present invention help to suppress, slow down, or prevent the formation of visible and / or invisible particles in preparations containing enveloped viruses. Preferably, the preparations containing enveloped viruses are substantially free of visible and / or invisible particles.

[0061] In another embodiment, a formulation of the present invention containing an enveloped virus preserves the infectivity of the enveloped virus and / or slows the decline in infectivity when stored at a temperature of 25°C.

[0062] In another embodiment, the preparation is useful for maintaining the infectivity titer and / or colloidal stability of enveloped viruses when stored at a temperature of 2 to 8°C.

[0063] In another embodiment, the preparation of the present invention containing an enveloped virus has visible and / or invisible particles that are below the limits set forth in the official standards.

[0064] In one embodiment relating to any of the above embodiments, the formulation containing the enveloped virus is a liquid formulation, a freeze-liquid formulation, or a dry formulation, such as a freeze-dried formulation.

[0065] In another embodiment, a liquid and / or frozen liquid formulation of the present invention containing an enveloped virus preserves the infectivity of the enveloped virus and / or slows the degradation of infectivity during storage at a temperature of 25°C for at least 28 days.

[0066] In another embodiment, preparations of the liquid and / or frozen liquid of the present invention containing an enveloped virus are useful for reducing the formation of visible and / or invisible particles during storage at a temperature of 25°C for up to 28 days.

[0067] In another embodiment, the liquid and / or frozen liquid formulations of the present invention containing enveloped viruses significantly inhibit the formation of particles of 2 μm or larger and / or 10 μm or larger during freeze-thaw stress.

[0068] In another embodiment, the liquid and / or frozen liquid formulations of the present invention containing enveloped viruses significantly inhibit the formation of particles of 2 μm or larger and / or 10 μm or larger after shaking stress.

[0069] In another embodiment, the liquid and / or frozen liquid formulations of the present invention containing an enveloped virus are useful for reducing, slowing, and / or preventing the formation of visible particles when stored at a temperature of 25°C. In a related embodiment, the liquid and / or frozen liquid formulations of the present invention containing an enveloped virus have concentrations of invisible particles of 2 μm or larger when stored at a temperature of 25°C and after one freeze-thaw cycle of less than 5000 particles / mL, less than 4000 particles / mL, less than 3000 particles / mL, less than 2000 particles / mL, or less than 1000 particles / mL. In further related embodiments, in the liquid and / or frozen liquid formulations of the present invention containing enveloped viruses, the concentration of invisible particles 10 μm or larger when stored at a temperature of 25°C and after one freeze-thaw cycle is less than 6,000 particles / container, less than 5,000 particles / container, less than 4,000 particles / container, less than 3,000 particles / container, less than 2,000 particles / container, less than 1,000 particles / container, less than 800 particles / container, less than 600 particles / container, less than 400 particles / container, less than 200 particles / container, or less than 100 particles / container.

[0070] In another embodiment, the preparation of the buffer solution using trisodium phosphate (Na3PO4) and / or phosphoric acid (H3PO4) while omitting sodium chloride reduces the molar osmotic pressure concentration, T g Increase the value.

[0071] In another embodiment, the preparation of a buffer solution using phosphoric acid (H3PO4) instead of hydrochloric acid reduces the molal osmotic pressure concentration in the preparation without using trehalose, T g Increase the value by 13°C to 19°C.

[0072] In another embodiment, the omission of sodium chloride eliminates the formation of chloride salts during freezing.

[0073] In relation to any of the above embodiments, the potency of a particular formulation is determined by the method shown in the examples, TCID 50 This can be tested by determining the particle. In relation to any of the above embodiments, the particle may be evaluated according to the method described in the example.

[0074] While we don't want to get bogged down in theory, it has been found that poloxamers and certain poloxamers 188 reduce the formation of aggregates and / or particles in liquid and / or frozen liquid oncolytic virus formulations, particularly after freeze / thaw cycles.

[0075] Furthermore, it was found that recombinant human albumin (rHA) could reduce the formation of aggregates and / or particles in the frozen liquid formulation, possibly through interaction with the virus surface. In addition, rHA was found to stabilize the infectivity of lyophilized viruses in the frozen liquid formulation during storage.

[0076] Surprisingly, it was also found that stress-induced particles in rHA-containing formulations, in liquid and / or frozen liquid formulations, can be effectively prevented by the addition of poloxamer 188.

[0077] The pharmaceutical preparations and compositions described herein are particularly useful for the preparation of enveloped viruses. According to the present invention, the term "enveloped virus" refers to any of the genus enveloped viruses capable of infecting humans, including but not limited to herpesviruses, poxviruses, orthomyxoviruses, paramyxoviruses, rhabdoviruses, and filoviruses. The term "rhabdovirus" as used herein refers to any enveloped virus of the genus capable of infecting humans, including but not limited to almendravirus, curiovirus, cytorhabdovirus, dichorhavirus, ephemerovirus, hapavirus, ledantevirus, lyssavirus, novirhabdovirus, nucleorhabdovirus, perhabdovirus, sigmavirus, spribivirus, sripuvirus, tibrovirus, tupavirus, varicosavirus, or becyclovirus.

[0078] Preferably, enveloped viruses are replicable. More preferably, oncolytic viruses and replicable oncolytic viruses. In this regard, the term oncolytic is used in its usual sense, which is well known in the art, and refers to a virus that can infect and lyse (destroy) cancer cells but cannot infect and lyse normal cells (to any significant degree). Preferably, oncolytic viruses can replicate within cancer cells. Oncolytic activity can be tested with various assay systems known to those skilled in the art (an exemplary in vitro assay is described in Muik et al., Cancer Res., 74(13), 3567-78, 2014). It has been found that oncolytic viruses can infect and lyse only certain types of cancer cells. Furthermore, the oncolytic effect may vary depending on the type of cancer cell. It has been found that oncolytic viruses are live viruses that can infect cancer cells and replicate within them.

[0079] In any embodiment, the enveloped virus may be a recombinant or non-recombinant enveloped virus, preferably a recombinant enveloped virus, preferably a recombinant enveloped virus belonging to the Rhabdoviridae family, more preferably a recombinant becyclovirus, and even more preferably a recombinant vesicular stomatitis virus.

[0080] The term “recombinant” refers to a virus, more specifically an enveloped virus, that contains an exogenous nucleic acid sequence inserted into its genome that is not naturally present in the parent virus. Thus, a recombinant virus refers to a nucleic acid or virus created by the artificial combination of two or more nucleic acid sequence segments of synthetic or semi-synthetic origin, linked to another nucleic acid in a permutation that is not naturally present or found in nature. The artificial combination is most commonly achieved by the artificial manipulation of isolated nucleic acid segments using well-established genetic engineering techniques. Generally, “recombinant” enveloped viruses as described herein refer to enveloped viruses created by standard genetic engineering techniques; for example, the enveloped viruses of the present invention are therefore genetically engineered or genetically modified enveloped viruses. Thus, the term “recombinant enveloped virus” includes enveloped viruses in which recombinant nucleic acids are stably incorporated into their genomes.

[0081] Characteristic features of members of the Rhabdoviridae family include mostly unsegmented, 10.8–16.1 kb negative-stranded single-stranded RNA, as well as genomes encoding at least five genes that encode structural proteins: nucleoproteins (N), large proteins (L), phosphorylated proteins (P), matrix proteins (M), and glycoproteins (G).

[0082] Becyclovirus species are primarily defined by serological methods combined with phylogenetic analysis of genomes. Biological characteristics such as host range and transmission mechanism are also used to distinguish virus species within the genus. Thus, the genus Becyclovirus forms a distinct monophyletic group, well supported by maximum likelihood phylogenetic trees inferred from complete L sequences.

[0083] Viruses assigned to different species within the genus Becyclovirus may have one or more of the following characteristics: A) a 20% difference in the minimum amino acid sequence in L; B) a 10% difference in the minimum amino acid sequence in N; C) a 15% difference in the minimum amino acid sequence in G; D) distinguishable by serological testing; and E) occupying different environmental niches, as evidenced by differences in the host and / or vector arthropods.

[0084] In a preferred embodiment, the vesicular stomatitis virus encodes at least the vesicular stomatitis virus nucleoprotein (N), macroprotein (L), phosphorylated protein (P), matrix protein (M), and glycoprotein (G) within its genome.

[0085] In a preferred embodiment, the vesicular stomatitis virus nucleoprotein (N) contains in its genome at least the amino acid sequence shown in SEQ ID NO: 1, or a functional variant that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 1, and contains the amino acid sequence shown in SEQ ID NO: 2, or a functional variant that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 2. The product encodes a phosphorylated protein (P), a large protein (L) containing a functional variant that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence shown in SEQ ID NO: 4, or a functional variant that is at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence shown in SEQ ID NO: 4.

[0086] It is understood by those skilled in the art that modifications to the sequences of the nucleoprotein (N), macroprotein (L), phosphorylated protein (P), matrix protein (M), or glycoprotein (G) of vesicular stomatitis virus can be carried out without impairing the basic function of such proteins. Such functional variants used herein retain all or part of their basic function or activity. For example, protein L is a polymerase and has an essential function in viral transcription and replication. Its functional variant must retain at least part of this ability. A clear indication of retention of basic function or activity is the successful generation of the virus (including these functional variants) that is still able to replicate in tumor cells and infect tumor cells. Tests of viral generation and infection and replication in tumor cells can be performed using various assay systems known to those skilled in the art (exemplary in vitro assays are described by Muik et al., Cancer Res., 74(13), 3567-78, 2014).

[0087] In a preferred embodiment, the vesicular stomatitis virus encodes at least the vesicular stomatitis virus nucleoprotein (N), macroprotein (L), phosphorylated protein (P), matrix protein (M), and glycoprotein (G) within its genome, wherein the macroprotein (L) contains an amino acid sequence having a sequence identity of 80% or more with SEQ ID NO: 3.

[0088] In a preferred embodiment, the vesicular stomatitis virus has a genome that encodes at least a vesicular stomatitis virus nucleoprotein (N), macroprotein (L), phosphorylated protein (P), matrix protein (M), and glycoprotein (G), wherein the nucleoprotein (N) contains an amino acid sequence having a sequence identity of 90% or more with SEQ ID NO: 1.

[0089] In a more preferred embodiment, the vesicular stomatitis virus encodes at least a vesicular stomatitis virus nucleoprotein (N), macroprotein (L), phosphorylated protein (P), matrix protein (M), and glycoprotein (G) within its genome, wherein the macroprotein (L) contains an amino acid sequence having a sequence identity of 80% or more with SEQ ID NO: 3, and the nucleoprotein (N) contains an amino acid sequence having a sequence identity of 90% or more with SEQ ID NO: 1.

[0090] Certain wild-type strains of vesicular stomatitis virus are known to be neurotoxic. Furthermore, infected individuals have been reported to rapidly initiate a potent humoral response with high antibody titers primarily directed towards glycoproteins. Neutralizing antibodies targeting glycoprotein G of vesicular stomatitis virus can limit viral spread and thus mediate protection against reinfection. However, viral neutralization limits the repeated application of the virus to cancer patients.

[0091] To eliminate these drawbacks, the wild-type glycoprotein G may be replaced with a glycoprotein derived from another virus. In this regard, glycoprotein replacement means (i) replacing the gene encoding wild-type glycoprotein G with the gene encoding glycoprotein GP of another virus, and / or (ii) replacing wild-type glycoprotein G with glycoprotein GP of another virus.

[0092] In a preferred embodiment, the enveloped virus is a vesicular stomatitis virus, and the glycoprotein G of the vesicular stomatitis virus is replaced with the glycoprotein GP of lymphocytic choriomeningitis virus (LCMV), preferably the glycoprotein of the WE-HPI strain. Such a vesicular stomatitis virus is described, for example, in International Publication No. 2010 / 040526 and named vesicular stomatitis virus-GP.

[0093] Therefore, in the most preferred embodiment, the enveloped virus is recombinant vesicular stomatitis virus, wherein the gene encoding glycoprotein G of vesicular stomatitis virus is replaced by the gene encoding glycoprotein GP of lymphocytic choriomeningitis virus, and / or, the glycoprotein G is replaced by glycoprotein GP of lymphocytic choriomeningitis virus.

[0094] In a preferred embodiment, the gene encoding the glycoprotein GP of lymphocytic choriomeningitis virus encodes a protein having an amino acid sequence as shown in SEQ ID NO: 5, or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with the amino acid sequence of SEQ ID NO: 5, while maintaining the functional characteristics of an enveloped virus containing glycoprotein GP encoding an amino acid sequence as shown in SEQ ID NO: 5.

[0095] It should be understood that recombinant enveloped viruses may also encode other cargo within their genome, such as tumor antigens, other chemokines, cytokines, or other immunomodulatory elements.

[0096] In a preferred embodiment, the RNA genome of the vesicular stomatitis virus includes or comprises sequences such as those shown in SEQ ID NO: 6, 7, or 8. Furthermore, the RNA genome of the vesicular stomatitis virus may also consist of or include such sequences, where the nucleic acids of the RNA genome are exchanged according to the degeneracy of the gene code without altering the respective amino acid sequences. In a further preferred embodiment, the RNA genome of the vesicular stomatitis virus includes or comprises coding sequences that are identical to or at least 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 6, 7, or 8.

[0097] definition In this specification, the terms “identical” or “identical ratio” in the context of two or more nucleic acid sequences or polypeptide sequences refer to two or more sequences or subsequences that, when compared and aligned to the greatest extent possible, have the same nucleotide residues or amino acid residues, or identical in a specific ratio. To determine the degree of identity, the sequences are aligned for optimal comparison (for example, gaps may be introduced within the first amino acid sequence or nucleic acid sequence for optimal alignment with the second amino acid sequence or nucleic acid sequence). The amino acid residues or nucleotides at the corresponding amino acid or nucleotide positions are then compared. If a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, the molecules are identical at that position. The degree of identity between two sequences is a function of the number of identical positions shared by the sequences (i.e., degree of identity % = number of identical positions / total number of positions (e.g., overlapping positions) × 100). In some embodiments, the two sequences being compared are of the same length after introducing gaps within the sequences as appropriate (e.g., extra sequences extending beyond the sequences being compared are excluded).

[0098] The determination of the degree of identity or similarity between two sequences can be achieved using mathematical algorithms. A preferred and non-restrictive example of a mathematical algorithm used for comparing two sequences is the algorithm from Karlin and Altschul, 1990, Proc. Natl. Acad. Sci. USA 87:2264-2268, as modified in Karlin and Altschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873-5877. Such algorithms are incorporated into the NBLAST and XBLAST programs of Altschul et al., 1990, J. Mol. Biol. 215:403-410. By performing a BLAST nucleotide search using the NBLAST program, score=100, and word length=12, homologous nucleotide sequences to the nucleic acid encoding the protein of interest can be obtained. By performing a BLAST protein search using the XBLAST program, score=50, and word length=3, homologous amino acid sequences for the protein of interest can be obtained. To obtain gapped alignments for comparison purposes, gapped BLAST can be used as described in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402. Alternatively, iterative searches can be performed using PSI-Blast to detect distant relationships between molecules (ibid.). When using the BLAST, gapped BLAST, and PSI-Blast programs, the default parameters of each program (e.g., XBLAST and NBLAST) can be used. Another preferred non-restrictive example of a mathematical algorithm used for sequence comparison is the algorithm of Myers and Miller, CABIOS (1989). Such algorithms are incorporated into the ALIGN program (version 2.0), which is part of the GCG sequence alignment software package.When using the ALIGN program to compare amino acid sequences, the PAM120 weight residue table, gap length penalty 12, and gap penalty 4 may be used. Additional algorithms for sequence analysis are known in the art and include ADVANCE and ADAM, as described in Torellis and Robotti, 1994, Comput. Appl. Biosci. 10:3-5; and FASTA, as described in Pearson and Lipman, 1988, Proc. Natl. Acad. Sci. USA 85:2444-8. Within FASTA, ktup is a control option that sets the sensitivity and speed of the search. When ktup=2, similar regions in the two sequences being compared are found by searching for aligned residue pairs; when ktup=1, single aligned amino acids are examined. ktup can be set to 2 or 1 for protein sequences, or to 1-6 for DNA sequences. If ktup is not specified, the default is 2 for proteins and 6 for DNA. Alternatively, the ordering of protein sequences can be performed using the CLUSTAL W algorithm, as described by Higgins et al., 1996, Methods Enzymol. 266:383-402.

[0099] The term "approximately" generally refers to the degree of acceptable error or deviation of a measured quantity, taking into account the nature or accuracy of the measurement. Typical exemplary degrees of error or deviation are within 5%, 3%, or 1% of a given number or range of numbers. For example, the expression "approximately 100" includes 105 and 95, or 103 and 97, or 101 and 99, and all numbers in between (e.g., 95.1, 95.2, etc. for the range of 95-105; or 97.1 or 97.2, etc. for the range of 97-103; or 99.1, 99.2, etc. for the range of 99-101). Unless otherwise specified, quantities shown herein are approximate, meaning that the term "approximately" can be inferred unless explicitly stated.

[0100] As used herein, the general embodiments of “contains” or “includes” encompass the more specific embodiment of “consistes of.” Furthermore, singular and plural forms are not used in a restrictive sense. As used herein, the singular forms “a,” “an,” and “the” refer to both singular and plural forms unless otherwise specified.

[0101] "Pharmaceutical preparation" or "compounding" refers to the process of producing a final pharmaceutical or drug by combining an active drug or active substance with a chemical substance, but also refers to the product of the process. Therefore, a final preparation refers to a pharmaceutical product such as a liquid, frozen liquid, dried preparation, or composition. Thus, in one embodiment, a pharmaceutical preparation is a pharmaceutical composition.

[0102] In this context, "pharmaceutical composition" refers to a dosage form that allows the biological activity of an active ingredient(s) to be clearly effective, and which contains no additional ingredients that are significantly toxic to the subject to whom the composition is to be administered, whether liquid, frozen, or dried. Such a composition is sterile.

[0103] A "dried preparation" or "dried pharmaceutical composition" is prepared by removing the liquid from a preparation containing an enveloped virus, which has been prepared in a liquid solution. Removal of the liquid can be achieved, for example, by evaporation, such as by applying the liquid solution to a solid substrate and evaporating the liquid, and / or by sublimation, such as by freeze-drying. The dried preparations / dried pharmaceutical compositions of the present invention are generally stored as dried preparations having a residual moisture content (RMC) of 0.5% to 10.0% (w / w). The dried preparations / dried pharmaceutical compositions can be reconstituted before administration in an aqueous solution (such as, but not limited to, sterile water, saline solution, aqueous dextrose, or glycerol). In certain embodiments, the dried preparations / dried pharmaceutical compositions of the present invention are stored as dried preparations containing a residual moisture content of 0.5% to 5% (w / w). In more specific embodiments, the dried preparations / dried pharmaceutical compositions of the present invention are stored as dried preparations containing a residual moisture content of 0.5% to 3% (w / w).

[0104] It will be understood that the dry pharmaceutical composition described in the present invention will first be prepared as a liquid pharmaceutical composition. Therefore, when a concentration range and / or pH range for a dry pharmaceutical composition is indicated, it will be further understood that the concentration range and / or pH range refers to the liquid pharmaceutical composition initially prepared before it is dried, and / or the liquid pharmaceutical composition obtained after the restoration of the dry pharmaceutical composition using an aqueous solution such as water.

[0105] In one embodiment, the pharmaceutical composition described in the present invention is a liquid pharmaceutical composition comprising an aqueous solution, preferably water, and any (dry) pharmaceutical composition as described herein.

[0106] Furthermore, in another embodiment, it can be seen that the pharmaceutical composition described in the present invention is a liquid pharmaceutical composition obtained by restoring a dry pharmaceutical composition in an aqueous solution, preferably in water.

[0107] It will be understood that the various formulations described herein may be provided as liquid, frozen liquid, and / or dried formulations. Therefore, unless otherwise specified, the general terms formulation or pharmaceutical formulation or composition or pharmaceutical composition encompass all liquid, frozen liquid, and / or dried compositions / formulations.

[0108] As used herein, the term "water" refers to water for injection.

[0109] "Pharmacologically acceptable" excipients (vehicles, additives) are suitable for parenteral administration to subjects.

[0110] In one embodiment, the pharmaceutical formulation of the present invention is stable.

[0111] "Stability" refers to chemical and physical stability, and can be qualitatively and / or quantitatively evaluated using various analytical techniques described in this technical field, for example, in Moving oncolytic viruses into the clinic: clinical-grade production, purification, and characterization of diverse oncolytic viruses. Mol Ther Methods Clin Dev. 2016 Apr 6;3:16018. doi: 10.1038 / mtm.2016.18. PMID: 27088104; PMCID: PMC4822647. Such methods include evaluating the formation of aggregates and particles (for example, by measuring turbidity, particles invisible to the naked eye, by light-blocking (LO) or microflow imaging (MFI = flow imaging microscopy (FIM), dynamic image analysis (DIA), and / or visual inspection of color and clarity) using high-performance size exclusion chromatography (HP-SEC); by evaluating charge heterogeneity using cation exchange chromatography (CEX) or capillary isoelectric focusing electrophoresis; by mass spectrometry; by capillary gel electrophoresis (CGE) analysis; by peptide mapping (e.g., digestion with trypsin or digestion with lysyl endopeptidase (Lys-C)) analysis; and by evaluating biological activity (infectivity). To measure stability, samples of the preparations of the present invention may be tested in a stability test, where the sample is exposed to stress conditions for a selected period and subsequently subjected to quantitative and qualitative analysis of chemical stability, physical stability, and infectivity using appropriate analytical techniques.

[0112] Therefore, stability can be measured by storing the sample at a selected temperature for a selected period, for example, up to 12 months, at various temperatures such as -80°C, -20°C, 2-8°C, or 25°C, and by using, for example, high-performance size exclusion chromatography, cation exchange chromatography, flow imaging microscopy, light shielding, or capillary isoelectric focusing, or infectivity can be measured by qualitative and quantitative analysis.

[0113] According to the above, a "stable formulation" is a formulation containing an enveloped virus that is physically and chemically stable and / or retains its biological activity during storage.

[0114] "Physical stability," in substance within the context of this invention, refers to an enveloped virus that shows little to no signs of agglutination, sedimentation, and / or reduction in infectivity. Methods for obtaining physical stability include, for example, size exclusion chromatography, light-blocking or microflow imaging, or dynamic image analysis (DIA), and visual inspection. In size exclusion chromatography, broadening or tailing of broad peaks may be considered a significant difference within the context of this invention under the test conditions, depending on the column used, operating pressure, and buffer flow rate. Using microflow imaging, a significant increase in the number of particles, especially particles larger than 2 μm and / or 10 μm, may be considered a significant difference, particularly if the number of particles exceeds the limits of the official standard. A method for obtaining infectivity is the 50% tissue culture infectious dose, i.e., the amount of cytopathogen that would cause cytopathic activity in 50% of seeded cells.

[0115] In the context of this invention, the terms “stress” or “stress conditions” refer to, for example, mechanical stress, thermal stress, photostress, or freeze-thaw and the resulting stress, as shown in particular in the Examples section. A variety of methods and conditions for stimulating mechanical stress, thermal stress, photostress, or stress resulting from freeze-thaw are known to those skilled in the art. Mechanical stress may be, for example, shaking at 300 rpm for up to 48 hours at room temperature, or carefully shaking a virus-containing vial manually. Thermal stress may refer to, for example, storage at a reduced or increased temperature over a period of time; in one example, the sample may be stored at temperatures of 5°C, 25°C, or 30°C, where 25°C and 30°C refer to accelerated stress conditions. Photostress may be, for example, storage of the sample at a light intensity of about 1100 lux over 5 days at various temperatures. The sample can be subjected to stress from freezing and thawing by repeatedly subjecting it to a cycle of freezing at a temperature of, for example, -80°C and thawing at room temperature for 2 hours, with this cycle being repeated 3 to 5 times.

[0116] The term "substantially chloride-free" means that no chloride ion source is added to the pharmaceutical composition, preferably no chloride ion source is added to the pharmaceutical composition from an external source. More preferably, the pharmaceutical composition is chloride ion-free.

[0117] buffer solution As used herein, “buffer solution” refers to a buffered solution that can withstand changes in pH due to the action of its acid-base conjugated components. As used herein, “pH” refers to the acidity or alkalinity of the composition at room temperature. Standard methods for measuring the pH of a composition are known to those skilled in the art. Typically, pH measurement involves calibrating the instrument, placing electrodes in a well-mixed sample, and then directly decoding the pH from a pH meter.

[0118] In various embodiments, the pharmaceutical composition may include a buffer. Exemplary buffers of the present invention include acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris.

[0119] sugar In various embodiments, the pharmaceutical composition may contain a sugar or a combination of several sugars. Exemplary sugars of the present invention include dextrose, fructose, galactose, glucose, raffinose, trehalose, or sucrose.

[0120] sugar alcohol In various embodiments, the pharmaceutical composition may contain a sugar alcohol or a combination of several sugar alcohols. Exemplary sugar alcohols of the present invention include mannitol, sorbitol, xylitol, maltitol, maltitol syrup, lactitol, inositol, glycerol, erythritol, isomalt, or hydrolyzed hydrogenated starch.

[0121] Poloxamer As used herein, "EO-PO block copolymer" means a copolymer consisting of blocks of poly(ethylene oxide) and poly(propylene oxide).

[0122] As used herein, "Pluronic®" is an EO (Registered Trademark) x -PO y -EO z This refers to the EO-PO block copolymer within the structure. This structure is also called a "poloxamer."

[0123] Regarding the general term poloxamer, these copolymers are typically named using a three-digit number following the letter P (for poloxamer): multiplying the first two digits by 100 gives the approximate molecular weight of the polyoxypropylene core, and multiplying the last digit by 10 gives the polyoxyethylene content (for example, P188 = a poloxamer with a polyoxypropylene molecular weight of 7680-9510 g / mol and a polyoxyethylene content of 20%). For poloxamer 188, the poly(propylene oxide) chain contains a number of units ranging from 25 to 30, and each poly(ethylene oxide) block consists of an average of 75-85 ethylene oxide units (L. Bollenbach, J. Buske, K. Mader, P. Garidel, International Journal of Pharmaceutics, Volume 620, 2022).

[0124] The "protein substance" used herein refers to albumin, gelatin, serum albumin, recombinant albumin, bovine serum albumin, porcine serum albumin, human serum albumin, recombinant human albumin, preferably human serum albumin (HSA) or recombinant human albumin (rHA).

[0125] In some embodiments, the pharmaceutical composition comprises human serum albumin (HSA), preferably recombinant human albumin (rHA). HSA is the most abundant protein found in human plasma. As used herein, the term “recombinant” means, in the context of “HA,” that rHA is either a genetically engineered product or produced by a recombinant production method. rHA is not derived from (isolated from or purified from) a natural product (e.g., human plasma), but can be produced, for example, through genetically engineered cells; however, other methods for obtaining rHA may be equally used by those skilled in the art.

[0126] All tables below should be read as indicating that the preparation contains or consists of components within the specified concentration range. Optional components may or may not be part of the preparation. Where the terms “at least one” or “one or more” are used with a concentration range, that range should be understood to apply individually to each component; for example, if there are two amino acids and only one concentration range of 1–300 mM is given, then each amino acid individually has a concentration range of 1–300 mM.

[0127] [Table 1]

[0128] [Table 2]

[0129] The formulations described in the present invention are shown below. Preferably, the formulations are provided as liquid or frozen liquid formulations.

[0130] [Table 3]

[0131] [Table 4]

[0132] [Table 5]

[0133] [Table 6]

[0134] [Table 7]

[0135] [Table 8]

[0136] [Table 9]

[0137] [Table 10]

[0138] [Table 11]

[0139] [Table 12]

[0140] [Table 13]

[0141] [Table 14]

[0142] [Table 15]

[0143] In a preferred embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 106 mM Trehalose, about 5 mg / ml rHA, and a pH of 7.0 to 8.0, preferably about 7.5.

[0144] In a preferred embodiment, the liquid or frozen liquid formulation contains an enveloped virus, about 9.75 mM Tris, about 146 mM Arginine, about 103 mM Trehalose, about 5 mg / ml rHA, and a pH of 7.0 to 8.0, preferably about 7.5.

[0145] In a preferred embodiment, before adding rHA and poloxamer 188, the liquid or frozen liquid preparation contains enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 200 mM Trehalose, and a pH of 7.0 to 8.0, preferably about 7.5. After adding rHA and poloxamer 188 to a final concentration of about 5 mg / mL, respectively, the liquid or frozen liquid preparation contains the following composition: enveloped virus, about 9.75 mM Tris, about 146 mM Arginine, about 195 mM Trehalose, about 5 mg / mL rHA, about 5 mg / mL Poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5.

[0146] In any of the above embodiments of the preparations, preferably liquid or frozen liquid preparations, the concentration of the buffer may be within any concentration range of 1 to 100 mM, for example, 1 to 90 mM, 1 to 80 mM, 1 to 70 mM, 1 to 60 mM, or 1 to 50 mM. Preferably, the concentration of the buffer is 5 mM to 50 mM, 5 mM to 40 mM, 5 mM to 30 mM, or 5 mM to 20 mM. Preferably, the preparation, preferably liquid or frozen liquid preparation, contains Tris buffer, more preferably Tris buffer at a concentration of 5 to 50 mM.

[0147] In any embodiment of the above formulations, preferably liquid or frozen liquid formulations, the concentration of the sugar is within any concentration range of 1 to 500 mM, for example, 1 to 490 mM, 1 to 480 mM, 1 to 470 mM, 1 to 460 mM, 1 to 450 mM, 1 to 440 mM, 1 to 430 mM, 1 to 420 mM, 1 to 410 mM, 1 to 400 mM, 1 to 390 mM, The concentrations may be 1-380 mM, 1-370 mM, 1-360 mM, 1-350 mM, 1-340 mM, 1-330 mM, 1-320 mM, 1-310 mM, 1-300 mM, 1-290 mM, 1-280 mM, 1-270 mM, 1-260 mM, 1-250 mM, 1-240 mM, 1-230 mM, 1-220 mM, 1-210 mM, or 1-200 mM. Preferably, the sugar concentration is 50 mM-300 mM, 50 mM-250 mM, or 50 mM-200 mM. Preferably, the preparation, preferably a liquid or frozen liquid preparation, contains sugars such as dextrose, fructose, galactose, glucose, raffinose, trehalose, or sucrose. More preferably, the sugar is trehalose, at a concentration of 50 to 250 mM.

[0148] In any of the above embodiments of the formulations, preferably liquid or frozen liquid formulations, the concentration of the amino acid is within any concentration range of 1 to 300 mM, for example, 1 to 290 mM, 1 to 280 mM, 1 to 270 mM, 1 to 260 mM, 1 to 250 mM, 1 to 240 mM, 1 to 230 mM, 1 to 220 mM, 1 to 210 mM, 1 to 200 mM, It can range from 1-190mM, 1-180mM, 1-170mM, 1-160mM, 1-150mM, 1-140mM, 1-130mM, 1-120mM, 1-110mM, 1-100mM, 1-90mM, 1-80mM, 1-70mM, 1-60mM, 1-50mM, 1-40mM, 1-30mM, 1-20mM, and 1-10mM. Preferably, the amino acid concentrations are 50mM-400mM, 50mM-290mM, 50mM-280mM, 50mM-270mM, 50mM-260mM, 50mM-250mM, 50mM-240mM, 50mM-230mM, 50mM-220mM, 50mM-210mM, 50mM-200mM, 50mM-190mM, 50mM-180mM, 50mM-170mM, 50mM-160mM, 50mM-150mM, 50mM-140mM, 50mM-130mM, 50mM-120mM, 50mM-110mM, and 50mM-100mM. Preferably, the preparation, preferably a liquid or frozen liquid preparation, contains an amino acid, such as arginine, alanine, phenylalanine, glycine, glutamine, glutamic acid, methionine, or lysine. More preferably, the amino acid is arginine. Most preferably, the concentration of arginine is 50-300 mM, 50-250 mM, or 50-200 mM.

[0149] In any embodiment of the above formulations, preferably liquid or frozen liquid formulations, the concentration of poloxamer is any concentration range from 0.01 g / L to 50 g / L, for example, 0.1 g / L to 50 g / L, 1 g / L to 50 g / L, 1.1 g / L to 50 g / L, 1.2 g / L to 50 g / L, 1.3 g / L to 50 g / L, 1.4 g / L to 50 g / L L, 1.5g / L~50g / L, 1.6g / L~50g / L, 1.7g / L~50g / L, 1.8g / L~50g / L, 1.9g / L~50g / L, 2.0g / L~5 It can be 0g / L, 2.1g / L~50g / L, 2.2g / L~50g / L, 2.3g / L~50g / L, 2.4g / L~50g / L, 2.5g / L~50g / L. More preferred ranges include 1 g / L to 45 g / L, 1 g / L to 40 g / L, 1 g / L to 35 g / L, 1 g / L to 30 g / L, 1 g / L to 25 g / L, 1 g / L to 20 g / L, 1 g / L to 15 g / L, 1.5 g / L to 10 g / L, 2.0 g / L to 10 g / L, or 2.5 g / L to 10 g / L. Preferably, the formulation, preferably a liquid or frozen liquid formulation, contains pharmaceutically acceptable poloxamer. More preferably, the poloxamer is poloxamer 188. Most preferred concentrations of poloxamer 188 are 1 g / L to 20 g / L, 1 g / L to 15 g / L, 1.5 g / L to 10 g / L, 2.0 g / L to 10 g / L, or 2.5 g / L to 10 g / L.

[0150] In any of the above formulations, preferably liquid or frozen liquid formulations, the concentration of the protein substance is any concentration range from 0.1 g / L to 50 g / L, for example, 1 g / L to 50 g / L, 1.1 g / L to 50 g / L, 1.2 g / L to 50 g / L, 1.3 g / L to 50 g / L, 1.4 g / L to 50 g / L, 1.5 g / L L~50g / L, 1.6g / L~50g / L, 1.7g / L~50g / L, 1.8g / L~50g / L, 1.9g / L~50g / L, 2.0g / L~50g / L, 2.1g / L~50g / L, 2.2g / L~50g / L, 2.3g / L~50g / L, 2.4g / L~50g / L, 2.5g / L~50g / L. More preferred ranges include 1 g / L to 45 g / L, 1 g / L to 40 g / L, 1 g / L to 35 g / L, 1 g / L to 30 g / L, 1 g / L to 25 g / L, 1 g / L to 20 g / L, 1 g / L to 15 g / L, 1.5 g / L to 10 g / L, 2.0 g / L to 10 g / L, or 2.5 g / L to 10 g / L. Preferably, the formulation, preferably a liquid or frozen liquid formulation, contains rHA. Most preferred concentrations of rHA are 1 g / L to 20 g / L, 1 g / L to 15 g / L, 1.5 g / L to 10 g / L, 2.0 g / L to 10 g / L, or 2.5 g / L to 10 g / L.

[0151] The pH of the above-described formulations, preferably liquid or frozen liquid formulations, is typically maintained within the range of 5 to 9, or 6 to 9, or 6.5 to 8.5, or 6.5 to 8.0, preferably 7.0 to 8.0.

[0152] In any of the above formulations, preferably in liquid or frozen liquid form, the enveloped virus is preferably becyclovirus, more preferably vesicular stomatitis virus, and most preferably vesicular stomatitis virus having glycoprotein G substituted with glycoprotein GP of lymphocytic choriomeningitis virus (LCMV). In any of the above formulations, preferably in liquid or frozen liquid form, the virus concentration is 1 × 10⁻⁶ 5 TCID 50 / mL ~ 1 × 10 12 TCID 50Any concentration range within / mL, or at least 1 × 10 5 TCID 50 / mL, at least 1 × 10⁻⁶ 6 TCID 50 / mL, at least 1 × 10⁻⁶ 7 TCID 50 / mL, at least 1 × 10⁻⁶ 8 TCID 50 / mL, or at least 1 × 10 9 TCID 50 The concentration can be as low as / mL. Other ranges are 1 × 10 6 TCID 50 / mL ~ 1 × 10 12 TCID 50 / mL, 1 × 10 6 TCID 50 / mL ~ 1 × 10 11 TCID 50 Includes / mL, etc.

[0153] Further formulations described in the present invention are shown below.

[0154] [Table 16]

[0155] [Table 17]

[0156] [Table 18]

[0157] [Table 19]

[0158] [Table 20]

[0159] Table 21

[0160] Table 22

[0161] Table 23

[0162] Table 24

[0163] Table 25

[0164] Table 26

[0165] Table 27

[0166] Table 28

[0167] Table 29

[0168] In its broadest form, the present invention relates to a pharmaceutical composition comprising an enveloped virus and a protein substance and / or a poly(ethylene oxide) / poly(propylene oxide) block copolymer. The phrase "at least one of a buffer, a sugar, or an amino acid" means that the formulation must contain at least one of such components, but may contain two or all three of such components. For example, a formulation containing at least one of a buffer, a sugar, or an amino acid may contain a buffer; sugar; amino acid; buffer and sugar; buffer and amino acid; buffer, sugar and amino acid; or sugar and amino acid. It will also be understood that the formulation may contain, for example, two or more amino acids, two or more sugars, etc. Preferably, the formulation is provided as a liquid or frozen liquid formulation.

[0169] In any of the above formulations, preferably in liquid or frozen liquid form, the poloxamer concentration is 0.01-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.01-40 g / L, 0.01~30g / L, 0.01~20g / L, 0.01~10g / L, 0.01~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0. 1~10g / L, 0.1~5g / L, 0.2~40g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0.3 ~30g / L, 0.3~20g / L, 0.3~10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4 ~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, The concentration can be in any of the following ranges: 1-10 g / L, 1-5 g / L, 2-40 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L. Preferably, the poloxamer is poloxamer 188, and the concentration is in the range of 0.1-10 g / L.

[0170] In any of the above formulations, preferably in the form of a liquid or frozen liquid formulation, the concentration of the protein substance is 0.05-50 g / L, 0.1-50 g / L, 0.2-50 g / L, 0.3-50 g / L, 0.4-50 g / L, 0.5-50 g / L, 1-50 g / L, 2-50 g / L, 3-50 g / L, 4-50 g / L, 5-50 g / L, 0.05- 40g / L, 0.05~30g / L, 0.05~20g / L, 0.05~10g / L, 0.05~5g / L, 0.1~40g / L, 0.1~30g / L, 0.1~20g / L, 0.1~10g / L, 0.1~5g / L, 0.2~40g / L, 0.2~30g / L, 0.2~20g / L, 0.2~10g / L, 0.2~5g / L, 0.3~40g / L, 0. 3~30g / L, 0.3~20g / L, 0.3~10g / L, 0.3~5g / L, 0.4~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0. 4~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5~10g / L, 0.5~5g / L, 1~40g / L, 1~30g / L, 1~20g / L, The concentration can be within any of the following ranges: 1-10 g / L, 1-5 g / L, 2-40 g / L, 2-30 g / L, 2-20 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L. Preferably, the protein substance is recombinant human albumin or human serum albumin, and its concentration is within the range of 0.5-10 g / L.

[0171] In any of the above formulations, preferably a liquid or frozen liquid formulation, the concentration of the buffer can be within any of the following concentration ranges: 1 to 100 mM, 1 to 90 mM, 1 to 80 mM, 1 to 70 mM, 1 to 60 mM, 1 to 50 mM, 1 to 40 mM, 1 to 30 mM, 1 to 20 mM, or 1 to 10 mM. Preferably, the formulation, preferably a liquid or frozen liquid formulation, contains Tris buffer, more preferably Tris buffer at a concentration of 5 to 50 mM.

[0172] In any of the above formulations, preferably liquid or frozen liquid formulations, the sugar concentration is 10-1000 mM, 10-900 mM, 10-800 mM, 10-700 mM, 10-600 mM, 10-500 mM, 10-400 mM, 10-300 mM, 10-200 mM, 20-1000 mM, 20-900 mM, 20-800 mM, 20-700 mM, 20-600 mM, 20-500 mM, 20-400 mM, 20-300 mM, 20-200 mM, 30-1000 mM, 30-900 mM, 30-800 mM The concentration can be any of the following ranges: mM, 30-700 mM, 30-600 mM, 30-500 mM, 30-400 mM, 30-300 mM, 30-200 mM, 40-1000 mM, 40-900 mM, 40-800 mM, 40-700 mM, 40-600 mM, 40-500 mM, 40-400 mM, 40-300 mM, 40-200 mM, 50-1000 mM, 50-900 mM, 50-800 mM, 50-700 mM, 50-600 mM, 50-500 mM, 50-400 mM, 50-300 mM, or 50-200 mM. Preferably, the concentration of the sugar is 50 mM to 300 mM, 50 mM to 250 mM, or 50 mM to 200 mM. Preferably, the preparation, preferably a liquid or frozen liquid preparation, contains a sugar, such as dextrose, fructose, galactose, glucose, raffinose, trehalose, or sucrose. More preferably, the sugar is trehalose and is within a concentration of 50 to 250 mM.

[0173] The pH of the above-described formulations, preferably liquid or frozen liquid formulations, is typically maintained within the range of 5 to 9, or 6 to 9, or 6 to 8, or 6.5 to 8.5, or 6.5 to 8.0, preferably 7.0 to 8.0.

[0174] In an embodiment of any of the above-described formulations, preferably a liquid or frozen liquid formulation, the enveloped virus preferably has a glycoprotein G replaced with the glycoprotein GP of a vesiculovirus, more preferably vesicular stomatitis virus, and most preferably lymphocytic choriomeningitis virus (LCMV). In an embodiment of any of the above-described formulations, preferably a liquid or frozen liquid formulation, the virus concentration is 1×10 5 of TCID 50 / mL to 1×10 12 of TCID 50 / mL within any concentration range, at least 1×10 5 of TCID 50 / mL, at least 1×10 6 of TCID 50 / mL, at least 1×10 7 of TCID 50 / mL, at least 1×10 8 of TCID 50 / mL, or at least 1×10 9 of TCID 50 / mL and can be at a concentration of. Other ranges include 1×10 6 of TCID 50 / mL to 1×10 12 of TCID 50 / mL, 1×10 6 of TCID 50 / mL to 1×10 11 of TCID 50 / mL and the like.

[0175] In a preferred embodiment, the liquid or frozen liquid formulation contains an enveloped virus, about 10 mM Tris, about 150 mM arginine, about 100 mM trehalose, about 0.5 mg / mL poloxamer 188, about 2 mg / ml rHA, pH 7.0 to 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

[0176] In a preferred embodiment, the liquid or frozen liquid formulation comprises an enveloped virus, about 8 - 12 mM of Tris, about 140 - 160 mM of arginine, about 90 - 110 mM of trehalose, about 0.4 - 0.6 mg / mL of poloxamer 188, about 1.5 - 2.5 mg / ml of rHA, pH 7.0 - 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

[0177] In another embodiment, the liquid or frozen liquid formulation comprises an enveloped virus, about 160 mM of arginine, about 0.5 mg / mL of poloxamer 188, pH 7.0 - 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0178] In another embodiment, the liquid or frozen liquid formulation comprises an enveloped virus, about 140 - 160 mM of arginine, about 0.4 - 0.6 mg / mL of poloxamer 188, pH 7.0 - 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0179] In another embodiment, the liquid or frozen liquid formulation comprises an enveloped virus, about 10 mM of Tris, about 150 mM of arginine, about 0.5 mg / mL of poloxamer 188, pH 7.0 - 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0180] In another embodiment, the liquid or frozen liquid formulation comprises an enveloped virus, about 8 mM - 12 mM of Tris, about 140 - 160 mM of arginine, about 0.4 - 0.6 mg / mL of poloxamer 188, pH 7.0 - 8.0, preferably about pH 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0181] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 150 mM arginine, about 0.5 mg / mL poloxamer 188, about 2 mg / mL rHA, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0182] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 140–160 mM arginine, about 0.4–0.6 mg / mL poloxamer 188, about 1.5–2.5 mg / mL rHA, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0183] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 10 mM Tris, about 150 mM arginine, about 0.5 mg / mL poloxamer 188, about 2 mg / mL rHA, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0184] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 8–12 mM Tris, about 140–160 mM arginine, about 0.4–0.6 mg / mL poloxamer 188, about 1.5–2.5 mg / mL rHA, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0185] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 150 mM arginine, about 100 mM trehalose, about 0.5 mg / mL poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0186] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 140–160 mM arginine, about 90–110 mM trehalose, about 0.4–0.6 mg / mL poloxamer 188, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0187] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 100 mM Trehalose, about 0.5 mg / mL Poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0188] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 8–12 mM Tris, about 140–160 mM Arginine, about 90–110 mM Trehalose, about 0.4–0.6 mg / mL Poloxamer 188, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0189] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 150 mM arginine, about 100 mM trehalose, about 2 mg / mL rHA, about 0.5 g / L poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0190] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 140–160 mM arginine, about 90–110 mM trehalose, about 1.5–2.5 mg / mL rHA, about 0.4–0.6 g / L poloxamer 188, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0191] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 100 mM Trehalose, about 2 mg / mL rHA, about 0.5 g / L Poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0192] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 8–12 mM Tris, about 140–160 mM Arginine, about 90–110 mM Trehalose, about 1.5–2.5 mg / mL rHA, about 0.4–0.6 g / L Poloxamer 188, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0193] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 8–12 mM Tris, about 140–160 mM Arginine, about 90–110 mM Trehalose, about 4–6 mg / mL rHA, and a pH of 7.0–8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0194] In another embodiment, the liquid or frozen liquid preparation contains an enveloped virus, about 10 mM Tris, about 150 mM Arginine, about 200 mM Trehalose, about 5 g / L rHA, about 5 g / L Poloxamer 188, and a pH of 7.0 to 8.0, preferably about 7.5. Preferably, the pH of the composition is adjusted using phosphoric acid.

[0195] [Table 30]

[0196] [Table 31]

[0197] In another embodiment, Tris in the buffer in any of the formulations shown in Table 4a may be omitted or replaced with any other buffer as described herein.

[0198] Further aspects In a second aspect, the present invention relates to a pharmaceutical composition comprising an enveloped virus, a buffer, amino acids, (optionally) sugars, and protein substances and / or a block copolymer of poly(ethylene oxide) and poly(propylene oxide).

[0199] In one embodiment relating to the second aspect, the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, or glutamine, and is preferably arginine.

[0200] In one embodiment relating to the second aspect, the composition is substantially chloride-free, preferably substantially sodium chloride-free.

[0201] In one embodiment relating to the second aspect, the pH of the composition is 5 to 9, or 6 to 9, or 6.5 to 8.5, or 6.5 to 8.0, preferably 7.0 to 8.0. In the related embodiment, the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

[0202] In one embodiment relating to the second aspect, the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, and is preferably Tris.

[0203] In one embodiment relating to the second aspect, the sugar is sucrose or trehalose, preferably trehalose.

[0204] In one embodiment relating to the second aspect, the composition further comprises one or more sugar alcohols. In the related embodiment, the one or more sugar alcohols(s) are selected from the group consisting of mannitol, sorbitol, xylitol, maltitol, maltitol syrup, lactitol, inositol, glycerol, erythritol, isomalt, or hydrolyzed hydrogenated starch. In a further related embodiment, the one or more sugar alcohols(s) are mannitol and / or sorbitol, preferably a combination of mannitol and sorbitol.

[0205] In one embodiment relating to the second aspect, the poly(ethylene oxide) and poly(propylene oxide) block copolymer is a poloxamer, preferably a pharmaceutically acceptable poloxamer, more preferably a poloxamer 188.

[0206] In one embodiment relating to the second aspect, the protein substance is albumin, gelatin, preferably human serum albumin, or recombinant human albumin.

[0207] In one embodiment relating to the second aspect, the composition comprises both a protein substance and a poly(ethylene oxide) / poly(propylene oxide) block copolymer. In the related embodiment, the protein substance is recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188.

[0208] In one embodiment or any of the embodiments relating to the second aspect, the pharmaceutical composition comprises an enveloped virus, a buffer solution at a concentration of 1 mM to 100 mM, arginine at a concentration of 10 mM to 500 mM, sugar at a concentration of 10 mM to 1000 mM, poloxamer 188 at a concentration of 0.01 g / L to 50 g / L, and / or recombinant human albumin at a concentration of 0.1 g / L to 50 g / L.

[0209] In one embodiment or any of the embodiments relating to the second aspect, the pharmaceutical composition comprises: an enveloped virus; a buffer solution selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, and Tris, preferably Tris buffer, at a concentration of 1 mM to 100 mM; arginine at a concentration of 10 mM to 500 mM; a sugar selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, or sucrose, preferably trehalose, at a concentration of 10 mM to 1000 mM; and poloxamer 188 at a concentration of 0.01 g / L to 50 g / L and / or recombinant human albumin at a concentration of 0.1 g / L to 50 g / L.

[0210] In a third aspect, the present invention relates to a pharmaceutical composition comprising an enveloped virus, Tris buffer, arginine, (optionally) a sugar, and poloxamer 188 and / or recombinant human albumin.

[0211] In one embodiment relating to the third aspect, the pharmaceutical composition comprises an enveloped virus, Tris buffer, arginine, (optionally) a sugar, and poloxamer 188, and recombinant human albumin.

[0212] In one embodiment relating to the third aspect, the poloxamer 188 is present in a concentration of 0.01 g / L to 50 g / L.

[0213] In one embodiment related to the third aspect, the recombinant human albumin is present in a concentration of 0.1 g / L to 50 g / L.

[0214] In a fourth aspect, the present invention relates to a pharmaceutical composition comprising: an enveloped virus; a buffer solution selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, and Tris, preferably Tris buffer, in a concentration of 1 mM to 100 mM; arginine in a concentration of 10 mM to 500 mM; a sugar selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, or sucrose, preferably trehalose, in a concentration of 10 mM to 1000 mM; and poloxamer 188 in a concentration of 0.01 g / L to 50 g / L and / or recombinant human albumin in a concentration of 0.1 g / L to 50 g / L.

[0215] In one embodiment relating to the fourth aspect, the pharmaceutical composition comprises an enveloped virus, Tris buffer at a concentration of 1 mM to 100 mM, arginine at a concentration of 10 mM to 500 mM, trehalose at a concentration of 10 mM to 1000 mM, and poloxamer 188 at a concentration of 0.01 g / L to 50 g / L, and / or recombinant human albumin at a concentration of 0.1 g / L to 50 g / L.

[0216] In one embodiment and a group of embodiments relating to any of the above embodiments, the enveloped virus is a Rhabdoviridae, preferably a becyclovirus or a vesicular stomatitis virus (VSV). In the relevant embodiments, the enveloped virus is recombinant vesicular stomatitis virus (VSV), wherein the gene encoding glycoprotein G of vesicular stomatitis virus is replaced by the gene encoding glycoprotein GP of lymphocytic choriomeningitis virus, and / or glycoprotein G is replaced by glycoprotein GP of lymphocytic choriomeningitis virus.

[0217] In one embodiment and a group of embodiments relating to any of the above embodiments, the pharmaceutical composition is a liquid or frozen liquid pharmaceutical composition. In the relevant embodiments, the composition is stored frozen at a temperature of approximately -80°C, -70°C, -60°C, -50°C, -40°C, -35°C, -30°C, -25°C, -20°C, -15°C, -10°C, or -5°C.

[0218] In a fifth aspect, the present invention relates to a product produced by freeze-drying a liquid pharmaceutical composition.

[0219] In a sixth aspect, the present invention relates to a dry pharmaceutical composition produced by a method comprising the step of removing water from a pharmaceutical composition, the composition comprising an enveloped virus, Tris buffer in a concentration of 1 mM to 100 mM, trehalose in a concentration of 10 mM to 1000 mM, arginine in a concentration of 10 mM to 500 mM, poloxamer 188 in a concentration of 0.01 g / L to 50 g / L, and / or recombinant human albumin in a concentration of 0.1 g / L to 50 g / L.

[0220] In one embodiment relating to the sixth aspect, a pharmaceutical composition containing ice is obtained by freezing the pharmaceutical composition, and then the water is removed. In a related embodiment, the method further includes the step of removing water by placing the liquid pharmaceutical composition in a vacuum under controlled temperature and pressure. In a further related embodiment, the method is freeze-drying. In another related embodiment, the dried pharmaceutical composition contains less than about (0.5% to 5%) w / w of water.

[0221] In another embodiment, the pharmaceutical composition comprises water and a product of the fifth embodiment, or water and a dry pharmaceutical composition of the sixth embodiment and any of its embodiments.

[0222] Examples method: Preparations and viruses to be tested The virus being tested is a vesicular stomatitis virus in which the wild-type glycoprotein G is replaced by the glycoprotein GP of lymphocytic choriomeningitis virus; such viruses are subsequently named VSV-GP (vesicular stomatitis virus-glycoprotein). In some cases, VSV-GP further codes for a cargo, and such viruses are subsequently named VSV-GP-cargo 1 / 2 / or 3. The viral material VSV-GP or VSV-GP-cargo 1 / 2 / or 3 is approximately 5 × 10⁻⁶. 9 TCID 50 It is used undiluted at a concentration of / ml. A dialysis cassette (Slide-a-Lyzer dialysis cassette, Thermo, MWCO (molecular weight cutoff) 10kDa, 12ml) is used to introduce the virus into the specific formulation. The sample is dialyzed three times for 2 hours each time, with slow agitation at a temperature of 2-8°C. The final step is performed overnight. Sterile filtration is performed using a 0.22μm polyethersulfone (PES) filter. For macromolecules that do not pass through the dialysis membrane, namely recombinant human albumin (rHA), dextran, and poloxamer 188, an appropriate volume of stock solution is added after dialysis. For the placebo sample, a certain amount of rHA and poloxamer 188 is added to 15ml each of the sterile-filtered formulation.

[0223] Infectivity (TCID) 50 ) Method for determining Cells and viruses: BHK (baby hamster kidney)-21 cells (603126 (C13), CLS) are cultured in 5% CO2 at 37°C. The culture medium (GMEM21710082, Thermo) is supplemented with 8.7% fetal bovine serum and 4.3% tryptose phosphate broth. The BHK-21 cells are washed with phosphate-buffered saline and detached from the cell culture flask by incubation with TrypLE® Select enzyme at 37°C for 6-8 minutes. The cells are picked up from the medium, counted using Flex2 (Nova Biomedical), and seeded into 96-well plates.

[0224] TCID50 assay: In a 96-well plate, seed 10 4 BHK-21 cells per well in 100 μl of supplemented Glasgow minimum essential medium (GMEM). After 24 hours of incubation, infect the adherent cells with an 11×0.5 log 10 serial dilution of the virus or only diluent (negative control), and then incubate for 3 days at a temperature of 37 °C and 5% carbon dioxide. Take a bright-field image of the well of the cell culture fluid using a Cytation5 multimode imaging reader (BioTek) with a 4× objective lens. Visually (by the naked eye) evaluate whether the cytopathic effect of the imaged well is positive or negative. The final titer [TCID 50 / ml] is calculated by the Spearman-Karber formula. For each virus sample, infection with serial dilutions is performed on a total of 8 plates on the same day. Based on such 8 replicates, TCID 50 / ml is calculated. As an assay control, a crude vesicular stomatitis virus collection stored at -80 °C is used and performed in separate 96-well plates at each time point.

[0225] Method for determining visible and invisible particles Detection of visible particles by visual inspection (VI) Visual inspection is carried out according to in-house standards in a stepwise procedure. The inspection is carried out by two trained inspectors.

[0226] Step 1: Before inspection, equilibrate the sample in the dark at room temperature.

[0227] Step 2: Place the vial outside the inspection zone on the workbench without rotating or inverting it. Inspect the bottom of the vial and the curved part of the vial for particles.

[0228] Step 3: Inspect the vial for sedimentation at the bottom of the vial, directly in front of the light source and outside the inspection zone. Inspect the vial in an upright position and handle it carefully to avoid causing any sediment to swirl up.

[0229] Step 4: Inspect the vial outside the inspection zone, directly in front of the light source. Keep the vial upright and rotating. If there is any sediment at the bottom of the vial, it will rise and form a sediment of particles invisible to the naked eye that may swirl around.

[0230] Step 5: Examine the vial for the presence of visible particles at 2,000–3,750 lux in accordance with the European Pharmacopoeia (9th edition; monograph 2.9.20). Homogenize the solution by gently rotating it to avoid the formation of bubbles, and examine the liquid for 5 seconds in front of a white background and 5 seconds in front of a black background within the examination area.

[0231] Step 6: As the final step, the vial is inspected outside the inspection zone, directly in front of the light source. The vial is carefully rotated. The sample is inspected for very small particles, which are invisible to the naked eye only if present in large numbers.

[0232] Detection of particles invisible to the naked eye using microfluidic imaging (MFI) Microfluid imaging measurements are performed using the MFI-5200 particle analyzer system equipped with a silane-coated, high-resolution 100 μm flow cell. Briefly, the sample is diluted 5-fold with ADB. A 0.25 ml pre-lane volume is followed by 0.6 ml of the sample. Between measurements, the flow cell is rinsed with water. Background irradiation is optimized by using water. Measurements are performed using MFI's MVSS software version 2-R5.0.0.43, and samples are analyzed using MFI MVAS software version 1.3.0.1007.

[0233] Freeze drying Samples in 2R vials (0.4 ml) with Flurotec® stoppers are freeze-dried using a pilot-scale LCD-2-6D freeze-dryer (Martin Christ Gefriertrocknungsanlagen GmbH, Osterohde, Germany). Vacuum during the freeze-drying process is controlled by a diaphragm vacuum gauge. A conservative freeze-drying process is applied based on the calculated solid content and the estimated glass transition temperature (Tg') of the maximum freeze-concentrated solution. During the freeze-drying process, the product temperature, shelf temperature, condenser temperature, and chamber pressure (diaphragm vacuum gauge and Pirani vacuum gauge) are monitored. The product temperature is controlled by eight Pt 100 The samples are monitored by sensors (one sensor per preparation), and these sensors are located in different vials in the center of the samples on the third shelf. At the end of the process, the stoppers of the vials are closed in a freeze-drying oven at a pressure of 600 millibars under a nitrogen atmosphere. After stoppering the vials, the chamber is aerated to atmospheric pressure using nitrogen, and the samples are removed. After removing the samples from the freeze-drying oven, they are crimped, labeled, and stored under their respective storage conditions for further analysis. The freeze-dried preparations are stored at a temperature of 25°C or 30°C in an ICH110 cabinet (Memmert GmbH & Co. KG, Schwabach, Germany).

[0234] Determination of glass transition temperature Glass transition temperature T g and T g The result is determined using a DSC (Differential Scanning Calorimeter) 214 polymer oven (Erich Netzsch GmbH & Co Holding KG, Selb, Germany). 2–10 mg of the freeze-dried product was weighed in an aluminum pot in a humidity-controlled glove box (approximately 8% relative humidity), and the aluminum pot was subsequently sealed. g The values ​​are analyzed using Netzsch Proteus analysis software. All measurements are performed twice, and the results are calculated as the mean ± standard deviation.

[0235] Preparation of frozen liquid samples The sample is frozen by placing it in a freezer at -70°C or -80°C using a CoolCell LX (BioCision LLC) (Larkspur, California). The temperature is recorded by monitoring it with a temperature logger. For the freeze / thaw cycle, the sample is removed from the freezer and placed in an ICH110 cabinet (Memmert GmbH & Co. KG, Schwabach, Germany) at a temperature of 25°C ± 2°C / 60% ± 5% relative humidity, or 30°C ± 2°C / 65% ± 5% relative humidity, respectively. The temperature cycle is repeated one, three, or five times.

[0236] Example 1: Initial screening formulation Summary: The objective of the initial formulation development activities is to increase infectivity (TCID). 50 The objective was to characterize various virus formulations (Tables 5a-c) by the effects of storage and freeze / thaw (F / T) cycles on the formulations, and by the influence on particle formation in formulations designed to be stored as liquid, freeze-liquid formulations, or dried, i.e., freeze-dried formulations. In this example, VSV-GP-Load 1 was tested. Hereinafter, WP3a formulation refers to a liquid formulation, WP3b formulation refers to a freeze-liquid formulation, and WP3d formulation refers to a freeze-dried formulation. The general formulations were the same as the initial liquid, freeze-liquid, or freeze-dried formulations.

[0237] [Table 32]

[0238] [Table 33]

[0239] [Table 34]

[0240] Infectivity titer and the effect of preserving the preparation at 5°C or 25°C on particles invisible to the naked eye in the liquid preparation (WP3a). Figures 1-4 All liquid formulations (except WP3a_03, which contains polysorbate at 0.02 g / L) showed reasonable stability during storage at 5°C for 28 days (Figure 1), with an average titer decrease of 0.2 log units during storage. Some formulations also showed a titer decrease of 0.2 to 2.1 log units at 25°C (Figure 2). The most stable formulation was WP3a_General, which contains rHA (at 5 g / L). All formulations contained very low levels of microscopic particles (SvPs). After storage at 5°C (Figure 3) or 25°C (Figure 4), the formulation with the lowest amount of microscopic particles larger than 10 μm was WP3a_03, which contains polysorbate.

[0241] The effect of freeze / thaw cycles on infectivity titer and invisible particles in liquid formulations (WP3a). Figures 5-6 When liquid formulations of WP3a were subjected to one or three freeze / thaw cycles at temperatures ranging from -80°C to +25°C (Table 6), three freeze / thaw cycles resulted in a greater decrease in infectivity titer than one freeze / thaw cycle. The most stable formulation was WP3a_General, which contained rHA. The formulation containing polysorbate (WP3a_03) showed the greatest decrease in titer (Figure 5), and therefore, despite the positive effect of polysorbate on particle formation, it was no longer considered a suitable excipient for formulation development. The negative effect of polysorbate on the stability of vesicular stomatitis virus likely stems from its effect on its phospholipid envelope, which is easily damaged by detergents. The freeze / thaw cycle increased the amount of particles invisible to the naked eye in all formulations, except for WP3a_General, which contains rHA for particles larger than 10 μM, and WP3a_03, which contains polysorbate for particles larger than 10 μM (Figure 6).

[0242] [Table 35]

[0243] Effect of the freeze / thaw cycle on infectivity titer and the formation of microscopic particles in the frozen liquid formulation (WP3b). Figures 7-10

[0244] When VSV-GP was circulated from -20°C or -80°C to +25°C one, three, or five times (Table 7), the most stable formulation was again WP3a_General containing recombinant human albumin, followed by the formulation containing poloxamer 188, regardless of whether the formulation was circulated from -20°C to +25°C (Figure 7) or from -80°C to +25°C (Figure 8). Only in these formulations did five freeze / thaw cycles result in no further decrease in infectivity compared to three freeze / thaw cycles. Furthermore, in these formulations, the number of microscopic particles larger than 10 μM did not increase with freeze / thaw cycles, whether the formulation was circulated from -20°C to +25°C (Figure 9) or from -80°C to +25°C (Figure 10). When circulated at -20°C compared to -80°C, the number of particles more than doubled. Given previous observations that cleaning agents such as polysorbate damaged the envelope of VSV-GP, causing an unacceptable decrease in infectivity titer, it was highly surprising that the cleaning agent poloxamer 188 did not have such an effect on the envelope and titer. In contrast, the addition of poloxamer 188 to the formulation was beneficial for both infectivity titer and suppression of the formation of particles invisible to the naked eye.

[0245] [Table 36]

[0246] The effect of storing the preparation at 25°C on infectivity titer and the formation of microscopic particles in the freeze-dried preparation (WP3d). Figures 11-12 The infectivity of all formulations was stable immediately after freeze-drying, but decreased almost linearly when stored at 25°C (Figure 11). All formulations contained very low levels of particles invisible to the naked eye. The formulations with the lowest amount of particles larger than 10 μm invisible to the naked eye (Figure 12) were, as expected, WP3d_General containing rHA and WP3d_03 containing polysorbate. Stable freeze-dried cakes were obtained from all freeze-dried formulations except WP3d_General. This was because, in this formulation, sucrose was the sugar component instead of trehalose, which was present in all the other freeze-dried formulations. The glass transition temperature of WP3d_General (T g Since the value was approximately 25°C, which is close to the storage temperature, sucrose was replaced with trehalose in the following experiment.

[0247] Overall, WP3a_General=WP3b_General=WP3d_General, which contain rHA, performed well under all stress conditions and served as a foundation for further development efforts of freeze-dried formulations. The composition was 20 mM Tris (titrated with NaOH), 150 mM sucrose, 50 mM mannitol, 50 mM sorbitol, 20 mM glutamic acid, and 5 g / L rHA (pH 7.4).

[0248] Example 2: Development of liquid formulations for VSV-GP (Step 1) Figures 13-16 In parallel with the general development activities described above, a frozen liquid formulation for VSV-GP was developed.

[0249] In the initial work package, the following formulations were tested:

[0250] [Table 37]

[0251] VSV-GP DP (dried preparation) was subjected to buffer exchange (by dialysis and addition) to create prototypes for eight different formulations. The effects of freeze / thaw cycles and incubation at 25°C for up to two weeks on infectivity and the formation of microscopic particles were analyzed.

[0252] All samples appeared turbid at all time points and under all conditions. Individual, macroscopic particles were detected randomly and could not be associated with any specific excipient. • Freeze / thaw stress was observed to increase the concentration of particles larger than 2 μm in all formulations after one freeze / thaw cycle (Figure 13), and to increase the concentration of particles larger than 10 μm in formulations that did not contain rHA or gelatin (Figure 14). Storage at 25°C for up to two weeks most significantly induced particles larger than 2 μm in three rHA-added or gelatin-added formulations, as well as in the placebo vial of the rHA-added formulation, measured 23 days after filling. The formation of particles induced by storage also occurred in the absence of the VSV-GP dry preparation, and the number of particles increased with the concentration of rHA, suggesting that the observed particles may be attributable to the presence of rHA rather than the VSV-GP dry preparation / rHA interaction. The formulations containing rHA showed the highest infectivity after storage (Figure 15), but all differences were within the assay variability.

[0253] In the second work package, the effect of rHA on infectivity titer was re-examined for VSV-GP as well.

[0254] For this purpose, formulations containing 10 mM Tris, 106 mM Trehalose, 150 mM Arginine, and 5 mg / ml rHA (pH 7.5) were tested against formulations containing the same excipients but without rHA. The formulations were stressed either by a freeze-thaw process or by storing them at 25°C for 1 or 2 weeks. The infectious virus titer of the VSV-GP samples was determined using TCID. 50 The determination was made using an assay. All samples were analyzed with n=3.

[0255] The infectivity titers of starting materials containing and without rHA were similar (Figure 16). Filtration (T0) and freeze / thaw stress (T-FT) did not reduce the infectivity titer, despite the presence of rHA. The decreasing trend in infectivity titer was determined during storage at 25°C and 60% relative humidity for one week (T-1w) or two weeks (T-2w). The decrease was more pronounced in rHA-free samples, which showed an infectivity titer reduction of more than 2 / 3 log10 at T-2w compared to T-FT.

[0256] Example 3: Development of liquid formulations for VSV-GP (Step 2) Figures 17-19 Based on the results of the first work package, the second work package (step 2 of work package 3) was initiated, and the following formulations were tested.

[0257] [Table 38]

[0258] The effects of a freeze / thaw cycle alone, or a freeze / thaw cycle followed by incubation at 25°C for up to two weeks, are assessed using MFI and TCID. 50 The analysis was performed using analytical tools.

[0259] Freeze / thaw stress increased the concentration of particles ≥2 μm (Figure 17) and ≥10 μm (Figure 18) in all formulations. The addition of rHA significantly inhibited the formation of such particles after one freeze / thaw cycle. Subsequent incubation of thawed rHA-added samples at 25°C for up to two weeks resulted in only a slight increase, if any, in the number of particles ≥2 μm. Formulations containing 2.5 mg / ml recombinant human albumin or 5 mg / ml rHA showed comparable results, with the number of particles ≥2 μm and ≥10 μm slightly lower when using 5 mg / ml. As already observed in tests using VSV-GP, the addition of trehalose slightly reduced the number of particles ≥2 μm after freeze / thaw. Direct comparison of formulations (pH adjusted using either hydrochloric acid or phosphoric acid) showed a reduced number of particles ≥10 μm when phosphoric acid was added instead of hydrochloric acid.

[0260] Incubation at 25°C following freeze / thaw stress reduced infectivity in all preparations. The addition of 5 mg / ml of rHA appeared to be protective (Figure 19).

[0261] Example 8: Development of liquid formulations for VSV-GP-Cargo 2 Figure 20 In parallel with the general development activities described above, a frozen liquid formulation was developed for VSV-GP-cargo 2 (another variant encoding a different cargo). Development studies using VSV-GP-cargo 2 were conducted to further optimize the formulation and to confirm the excipients of the preliminary lead formulations. Development of the frozen liquid formulation was initiated with a formulation containing 5 mg / mL of rHA titrated to pH 7.5 with 9.75 mM Tris, 146 mM L-arginine, 103 mM Trehalose, and orthophosphate.

[0262] The following formulations were tested in different work packages.

[0263] Table 39

[0264] Table 40

[0265] Table 41

[0266] Table 42

[0267] Table 43

[0268] result:

[0269] Table 44 TIFF2026517175000045.tif161170

[0270] Table 45 TIFF2026517175000047.tif54170

[0271] In the first screening test, poloxamer 188 at a concentration of 5 mg / mL was shown to reduce the level of microscopic particles in the sample immediately after preparation, after freeze-thawing, after holding time (1 and 2 weeks at 25°C and 37°C), and after stirring stress (Table 15). Furthermore, in a placebo solution containing rHA but no poloxamer 188, a large number of microscopic particles were observed after 2 weeks at 25°C, while a small number of microscopic particles were observed in the same formulation containing 5 mg / mL of poloxamer 188. Interestingly, stress-induced particles in the rHA-containing formulation could be effectively prevented by the addition of poloxamer 188.

[0272] The presence of rHA is TCID 50 As shown, it stabilized the virus solution when stored at a temperature of 25°C (Figure 20). A similar stabilizing effect of rHA was observed in WP4A, but here, TCID 50 As shown by the analysis (data not shown), it stabilized the virus when stored at a temperature of 25°C for one month.

[0273] The decrease in arginine concentration from 150 mM to 50 mM (in the absence of rHA) negatively affected mechanical stress and retention time (1 month at 25°C) stability.

[0274] The stabilizing effect of poloxamer 188, as previously observed, was investigated at two different poloxamer 188 concentrations: 2.5 mg / mL and 5 mg / mL. Samples containing the higher concentration of poloxamer 188 (5 mg / mL) showed lower concentrations of invisible particles larger than 2 μm after mechanical stress. These samples also showed lower concentrations of particles larger than 2 μm and larger than 10 μm after circulation at temperatures ranging from 2–8°C to -80°C.

[0275] The effect of pH on viral stability in the presence of rHA and poloxamer 188 was further investigated in other work packages. The following pH values ​​were examined: pH 6.5, pH 7.0, pH 7.5, and pH 7.8. Virus solutions at lower pH levels (i.e., pH 6.5 and 7.0) showed higher levels of microscopic particles after storage at 25°C for one month (Table 16). Furthermore, these samples showed a slight reduction in viral particles by NTA (nanoparticle tracking analysis) after mechanical stress and freeze-thaw cycling. TCID 50 The decrease was observed in all samples stored at 25°C for one month.

[0276] This decrease was most pronounced for candidate formulations with the highest pH (i.e., pH 7.8) and lowest pH (i.e., pH 6.5). Further experiments using pH 7.0 and 7.3 confirmed the quality of stable dry preparations with pH 7.3 and pH 7.5.

[0277] Based on the results above prior to the addition of rHA and poloxamer 188, the preferred virus solution consists of 10 mM Tris, 150 mM L-arginine, and 200 mM trehalose (pH 7.5) (adjusted with phosphoric acid). After adding rHA and poloxamer 188 until the final concentration of each reaches 5 mg / mL, the preparation has the following composition: 9.75 mM Tris, 146 mM L-arginine, 195 mM trehalose, 5 mg / mL rHA, and 5 mg / mL poloxamer 188 (pH 7.5) (adjusted with orthophosphate).

[0278] Example 9: A formulation test is conducted by using an "experimental design" approach to test formulations. Decoded value: The formulations were tested in stability tests. All formulations containing L-arginine, poloxamer 188, and recombinant human albumin (rHA) were stable over a wide range of concentrations. Trehalose did not act as a stabilizer, but it did act as an osmotic regulator. Formulations without poloxamer 188 were less stable than those containing poloxamer 188.

[0279] [Table 46] TIFF2026517175000049.tif21170

Claims

1. A pharmaceutical composition comprising an enveloped virus, a protein substance, and / or a poly(ethylene oxide) / poly(propylene oxide) block copolymer.

2. The pharmaceutical composition according to claim 1, wherein the block copolymer of poly(ethylene oxide) and poly(propylene oxide) is a poloxamer.

3. The pharmaceutical composition according to claim 1, wherein the block copolymer of poly(ethylene oxide) and poly(propylene oxide) is poloxamer 188.

4. The pharmaceutical composition according to claim 1, wherein the protein substance is albumin or gelatin.

5. The pharmaceutical composition according to claim 1, wherein the protein substance is human serum albumin or recombinant human albumin.

6. The pharmaceutical composition according to claim 1, wherein the aforementioned block copolymer of poly(ethylene oxide) and poly(propylene oxide) is poloxamer 188, and the protein substance is human serum albumin or recombinant human albumin.

7. The pharmaceutical composition according to claim 6, which contains poloxamer 188 at a concentration of 0.01 to 50 g / L, 0.1 to 50 g / L, 0.2 to 50 g / L, 0.3 to 50 g / L, 0.4 to 50 g / L, 0.5 to 50 g / L, 1 to 50 g / L, 2 to 50 g / L, 3 to 50 g / L, 4 to 50 g / L, 5 to 50 g / L, 0.01 to 40 g / L, 0.01 to 30 g / L, 0.01 to 20 g / L, 0.01 to 10 g / L, 0.01 to 5 g / L, 0.1 to 40 g / L, 0.1 to 30 g / L, 0.1 to 20 g / L, 0.1 to 10 g / L, 0.1 to 5 g / L, 0.2 to 40 g / L, 0.2 to 30 g / L, 0.2 to 20 g / L, 0.2 to 10 g / L, 0.2 to 5 g / L, 0.3 to 40 g / L, 0.3 to 30 g / L, 0.3 to 20 g / L, 0.3 to 10 g / L, 0.3 to 5 g / L, 0.4 to 40 g / L, 0.4 to 30 g / L, 0.4 to 20 g / L, 0.4 to 10 g / L, 0.4 to 5 g / L, 0.5 to 40 g / L, 0.5 to 30 g / L, 0.5 to 20 g / L, 0.5 to 10 g / L, 0.5 to 5 g / L, 1 to 40 g / L, 1 to 30 g / L, 1 to 20 g / L, 1 to 10 g / L, 1 to 5 g / L, 2 to 40 g / L, 2 to 30 g / L, 2 to 20 g / L, 2 to 10 g / L, 3 to 40 g / L, 3 to 30 g / L, 3 to 20 g / L, 3 to 10 g / L, 4 to 40 g / L, 4 to 30 g / L, 4 to 20 g / L, 4 to 10 g / L, 5 to 40 g / L, 5 to 30 g / L, 5 to 20 g / L, or 5 to 10 g / L.

8. 0.05-50g / L, 0.1-50g / L, 0.2-50g / L, 0.3-50g / L, 0.4-50g / L, 0.5-50g / L, 1-50g / L, 2-50g / L , 3-50g / L, 4-50g / L, 5-50g / L, 0.05-40g / L, 0.05-30g / L, 0.05-20g / L, 0.05-10g / L, 0.05-5g / L, 0.1-40g / L, 0.1-30g / L, 0.1-20g / L, 0.1-10g / L, 0.1-5g / L, 0.2-40g / L, 0.2-30g / L, 0.2-20g / L, 0.2-10g / L, 0.2-5g / L, 0.3-40g / L, 0.3-30g / L, 0.3-20g / L, 0.3-10g / L, 0.3-5g / L, 0.4 ~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5-10g / L, 0.5-5g / L, 1-40g / L, 1-30g / L, 1-20g / L, 1-10g / L, 1-5g / L, 2-40g / L, 2-30g / L, 2-2 The pharmaceutical composition according to claim 6, comprising human serum albumin or recombinant human albumin at concentrations of 0 g / L, 2-10 g / L, 3-40 g / L, 3-30 g / L, 3-20 g / L, 3-10 g / L, 4-40 g / L, 4-30 g / L, 4-20 g / L, 4-10 g / L, 5-40 g / L, 5-30 g / L, 5-20 g / L, or 5-10 g / L.

9. A pharmaceutical composition according to any one of claims 1 to 8, further comprising at least one of an amino acid, a buffer solution, or a sugar.

10. The pharmaceutical composition of claim 9, wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, and is preferably arginine.

11. The pharmaceutical composition of claim 9, wherein the buffer solution is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, and is preferably Tris.

12. The pharmaceutical composition of claim 9, wherein the aforementioned sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, and is preferably trehalose.

13. The pharmaceutical composition according to any one of claims 1 to 12, further comprising one or more sugar alcohols.

14. The pharmaceutical composition of claim 13, wherein one or more sugar alcohols are selected from the group consisting of mannitol, sorbitol, xylitol, maltitol, maltitol syrup, lactitol, inositol, glycerol, erythritol, isomalt, and hydrolyzed hydrogenated starch.

15. The pharmaceutical composition of claim 14, wherein one or more sugar alcohols are mannitol and / or sorbitol, preferably a combination of mannitol and sorbitol.

16. The pharmaceutical composition according to claim 9 or 11, wherein the buffer solution has a concentration of 1 to 100 mM, 1 to 90 mM, 1 to 80 mM, 1 to 70 mM, 1 to 60 mM, 1 to 50 mM, 1 to 40 mM, 1 to 30 mM, 1 to 20 mM, or 1 to 10 mM.

17. The aforementioned sugars are 10-1000 mM, 10-900 mM, 10-800 mM, 10-700 mM, 10-600 mM, 10-500 mM, 10-400 mM, 10-300 mM, 10-200 mM, 20-1000 mM, 20-900 mM, 20-800 mM, 20-700 mM, 20-600 mM, 20-500 mM, 20-400 mM, 20-300 mM, 20-200 mM, 30-1000 mM, 30-900 mM, 30-800 mM, 30-700 mM, 30-600 mM, 30-5 A pharmaceutical composition according to claim 9 or 12, having a concentration of 00 mM, 30-400 mM, 30-300 mM, 30-200 mM, 40-1000 mM, 40-900 mM, 40-800 mM, 40-700 mM, 40-600 mM, 40-500 mM, 40-400 mM, 40-300 mM, 40-200 mM, 50-1000 mM, 50-900 mM, 50-800 mM, 50-700 mM, 50-600 mM, 50-500 mM, 50-400 mM, 50-300 mM, or 50-200 mM.

18. The pharmaceutical composition according to any one of claims 1 to 17, wherein the composition is substantially free of chlorides, preferably substantially free of sodium chlorides.

19. A pharmaceutical composition according to any one of claims 1 to 18, wherein the pH of the composition is 5 to 9, or 6 to 9, or 6.5 to 8.5, or 6.5 to 8.0, preferably 7.0 to 8.

0.

20. The pharmaceutical composition according to claim 19, wherein the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

21. - Enveloped viruses, - At least one of a buffer, an amino acid, or a sugar, and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

22. - Enveloped viruses, - Amino acids and buffer solutions, and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

23. - Enveloped viruses, - Amino acids and sugars, and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

24. - Enveloped viruses, - Amino acids, buffer solutions, and sugars, and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

25. - Enveloped viruses, - Buffer solution and sugar, and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

26. - Enveloped viruses, - A buffer, at least one of an amino acid or a sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate and Tris, preferably Tris), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

27. - Enveloped viruses, - Amino acids and buffer (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

28. - Enveloped viruses, - Amino acids, buffers, and sugars (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

29. - Enveloped viruses, - Buffer and sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate and Tris, preferably Tris), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

30. - Enveloped viruses, - A buffer, at least one of an amino acid or a sugar (wherein the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

31. - Enveloped viruses, - Amino acids and buffer solutions (wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

32. - Enveloped viruses, - Amino acids and sugars (wherein sugars are selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

33. - Enveloped viruses, - Amino acids and buffers and sugars (wherein sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

34. - Enveloped viruses, - Buffer solution and sugar (wherein sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

35. - Enveloped viruses, - A buffer solution, at least one of an amino acid or a sugar (wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

36. - Enveloped viruses, - Amino acids and sugars (wherein amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

37. - Enveloped viruses, - Amino acids, buffer solutions, and sugars (wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

38. - Enveloped viruses, - At least one of a buffer, an amino acid, or a sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

39. - Enveloped viruses, - Amino acids and buffers (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

40. - Enveloped viruses, - Amino acids and sugars (wherein amino acids are selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein sugars are selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

41. - Enveloped viruses, - Amino acids, buffer solutions, and sugars (wherein the buffer solution is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate, and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine, and glutamine, preferably arginine; wherein the sugar is selected from the group consisting of dextrose, fructose, galactose, glucose, raffinose, trehalose, and sucrose, preferably trehalose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

42. - Enveloped viruses, - Buffer and sugar (wherein the buffer is selected from the group consisting of acetate, citrate, histidine, succinate, HEPES, tartrate, phosphate, citrate / phosphate, lactate and Tris, preferably Tris; wherein the amino acid is selected from the group consisting of alanine, arginine, phenylalanine, glutamic acid, glycine, methionine, lysine and glutamine, preferably arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

43. - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

44. - Enveloped viruses, - Amino acids (where the amino acid is arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

45. - Enveloped viruses, - Sugars (where sugar is trehalose or sucrose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

46. - Enveloped viruses, - Amino acids (in this case, the amino acid is arginine), - Sugars (where sugar is trehalose or sucrose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

47. - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Sugars (where sugar is trehalose or sucrose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

48. - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Amino acids (where the amino acid is arginine), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

49. - Enveloped viruses, - Buffer solution (the buffer solution here is Tris), - Amino acids (in this case, the amino acid is arginine), - Sugars (where sugar is trehalose or sucrose), and - Protein material and / or poly(ethylene oxide) / poly(propylene oxide) block copolymer (wherein the protein material is selected from human serum albumin or recombinant human albumin, and the poly(ethylene oxide) / poly(propylene oxide) block copolymer is poloxamer 188) The pharmaceutical composition according to claim 1, comprising:

50. The pharmaceutical composition according to any one of claims 21, 22, 24-27, 29-32, 34-36, 38-40, 42-44, and 48-49, wherein the buffer solution has a concentration of 1-100 mM, 1-90 mM, 1-80 mM, 1-70 mM, 1-60 mM, 1-50 mM, 1-40 mM, 1-30 mM, 1-20 mM, or 1-10 mM.

51. The aforementioned sugars are found in concentrations of 10-1000 mM, 10-900 mM, 10-800 mM, 10-700 mM, 10-600 mM, 10-500 mM, 10-400 mM, 10-300 mM, 10-200 mM, 20-1000 mM, 20-900 mM, 20-800 mM, 20-700 mM, 20-600 mM, 20-400 mM, 20-300 mM, 20-200 mM, 30-1000 mM, 30-900 mM, 30-800 mM, 30-700 mM, 30-600 mM, 30-500 mM, 30-400 mM, and 30-300 mM. A pharmaceutical composition according to any one of claims 21, 23-25, 28-31, 33-39, 41-43, 46-47, 49, and 50, having a concentration of M, 30-200 mM, 40-1000 mM, 40-1000 mM, 40-900 mM, 40-900 mM, 40-800 mM, 50-700 mM, 50-600 mM, 50-500 mM, 50-400 mM, 50-300 mM, or 50-200 mM.

52. 0.01-50g / L, 0.1-50g / L, 0.2-50g / L, 0.3-50g / L, 0.4-50g / L, 0.5-50g / L, 1-50g / L, 2-50g / L , 3-50g / L, 4-50g / L, 5-50g / L, 0.01-40g / L, 0.01-30g / L, 0.01-20g / L, 0.01-10g / L, 0.01-5g / L, 0.1-40g / L, 0.1-30g / L, 0.1-20g / L, 0.1-10g / L, 0.1-5g / L, 0.2-40g / L, 0.2-30g / L, 0.2-20g / L, 0.2-10g / L, 0.2-5g / L, 0.3-40g / L, 0.3-30g / L, 0.3-20g / L, 0.3-10g / L, 0.3-5g / L, 0.4 ~40g / L, 0.4~30g / L, 0.4~20g / L, 0.4~10g / L, 0.4~5g / L, 0.5~40g / L, 0.5~30g / L, 0.5~20g / L, 0.5-10g / L, 0.5-5g / L, 1-40g / L, 1-30g / L, 1-20g / L, 1-10g / L, 1-5g / L, 2-40g / L, 2-30g / L, 2- A pharmaceutical composition according to any one of claims 21 to 51, comprising poloxamer 188 at concentrations of 20 g / L, 2 to 10 g / L, 3 to 40 g / L, 3 to 30 g / L, 3 to 20 g / L, 3 to 10 g / L, 4 to 40 g / L, 4 to 30 g / L, 4 to 20 g / L, 4 to 10 g / L, 5 to 40 g / L, 5 to 30 g / L, 5 to 20 g / L, or 5 to 10 g / L.

53. 0.05-50g / L, 0.1-50g / L, 0.2-50g / L, 0.3-50g / L, 0.4-50g / L, 0.5-50g / L, 1-50g / L, 2-50g / L, 3-50g / L, 4-50g / L, 5-50g / L, 0.05-40g / L, 0.05-30g / L, 0.05-20g / L, 0.05-10g / L, 0.05-5g / L, 0.1-40g / L, 0.1-30g / L, 0.1-20g / L, 0.1-10g / L, 0.1-5g / L, 0.2-40g / L, 0.2-30g / L, 0.2-20g / L , 0.2-10g / L, 0.2-5g / L, 0.3-40g / L, 0.3-30g / L, 0.3-20g / L, 0.3-10g / L, 0.3-5g / L, 0.4-40g / L , 0.4-30g / L, 0.4-20g / L, 0.4-10g / L, 0.4-5g / L, 0.5-40g / L, 0.5-30g / L, 0.5-20g / L, 0.5-10g / L, 0.5-5g / L, 1-40g / L, 1-30g / L, 1-20g / L, 1-10g / L, 1-5g / L, 2-40g / L, 2-30g / L, 2-20g / L, 2-1 A pharmaceutical composition according to any one of claims 21 to 51, comprising human serum albumin or recombinant human albumin at a concentration of 0 g / L, 3 to 40 g / L, 3 to 30 g / L, 3 to 20 g / L, 3 to 10 g / L, 4 to 40 g / L, 4 to 30 g / L, 4 to 20 g / L, 4 to 10 g / L, 5 to 40 g / L, 5 to 30 g / L, 5 to 20 g / L, or 5 to 10 g / L.

54. - Enveloped viruses, - Tris at approximately 1 to 100 mM, - Approximately 10-500 mM arginine, - Trehalose at approximately 10-500 mM, - Poloxamer 188 at approximately 0.1 to 5 mg / ml, - Recombinant human albumin at approximately 0.5 to 10 mg / ml, and -pH about 6-8 A pharmaceutical composition according to claim 1, comprising:

55. - Enveloped viruses, - Approximately 10 mM Tris, - Approximately 150 mM arginine, - Approximately 100 mM trehalose, - Approximately 0.5 mg / ml of poloxamer 188, - Approximately 2 mg / ml of recombinant human albumin, and -pH about 7.5 A pharmaceutical composition according to claim 54, comprising:

56. The pharmaceutical composition according to claim 54 or 55, wherein the pH of the composition is adjusted using phosphoric acid or sodium phosphate.

57. The pharmaceutical composition according to any one of claims 1 to 56, wherein the enveloped virus is of the Rhabdoviridae family, preferably becyclovirus or vesicular stomatitis virus (VSV).

58. The pharmaceutical composition according to claim 57, wherein the enveloped virus is recombinant vesicular stomatitis virus (VSV), and the gene encoding glycoprotein G of the vesicular stomatitis virus is replaced by the gene encoding glycoprotein GP of lymphocytic choriomeningitis virus (LCMV), and / or the glycoprotein G is replaced by glycoprotein GP of LCMV.

59. The pharmaceutical composition has at least 1×10 ５ TCID ５０ / mL, at least 1×10 ６ TCID ５０ / mL, at least 1×10 ７ TCID ５０ / mL, at least 1×10 ８ TCID ５０ / mL, at least 1×10 ９ TCID ５０ / mL, or at least 1×10 ９ TCID ５０ / mL concentration of an enveloped virus, preferably a vesiculovirus or vesicular stomatitis virus (VSV), the pharmaceutical composition according to claim 57 or 58.

60. The aforementioned pharmaceutical composition is 1 × 10 ５ TCID ５０ / mL ~ 1 x 10 １２ TCID ５０ / mL, 1 x 10 ６ TCID ５０ / mL ~ 1 x 10 １２ TCID ５０ / mL, 1 x 10 ７ TCID ５０ / mL ~ 1 x 10 １２ TCID ５０ / mL, 1 x 10 ８ TCID ５０ / mL ~ 1 x 10 １２ TCID ５０ / mL, 1 x 10 ５ TCID ５０ / mL ~ 1 x 10 １１ TCID ５０ / mL, 1 x 10 ５ TCID ５０ / mL ~ 1 x 10 １０ TCID ５０ / mL, or 1 × 10 ５ TCID ５０ / mL ~ 1 x 10 ９ TCID ５０ The pharmaceutical composition according to claim 57 or 58, comprising an enveloped virus, preferably becyclovirus or vesicular stomatitis virus (VSV), in a concentration range of / mL.

61. The pharmaceutical composition according to any one of claims 1 to 60, wherein the pharmaceutical composition is a liquid or a frozen liquid pharmaceutical composition.

62. The liquid pharmaceutical composition according to claim 61, wherein the composition is stored frozen at a temperature of approximately -80°C, -70°C, -60°C, -50°C, -40°C, -35°C, -30°C, -25°C, -20°C, -15°C, -10°C, or -5°C.

63. A product produced by freeze-drying the liquid pharmaceutical composition of claim 62.

64. A dried pharmaceutical composition produced by a method comprising the step of removing water from the pharmaceutical composition according to any one of claims 1 to 63.

65. The dried pharmaceutical composition according to claim 64, wherein the pharmaceutical composition is frozen to obtain a pharmaceutical composition containing ice, and then water is removed.

66. The dry pharmaceutical composition according to claim 65, wherein the method further comprises the step of removing water from the liquid pharmaceutical composition by placing it in a vacuum under controlled temperature and pressure.

67. A dried pharmaceutical composition according to any one of claims 64 to 66, wherein the method is freeze-drying.

68. A dry pharmaceutical composition according to any one of claims 64 to 67, containing less than approximately (0.5% to 5%) w / w of water.

69. A pharmaceutical composition comprising water and the product of claim 61 or a dried pharmaceutical composition of any one of claims 62 to 66.

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