Composition of an antiviral agent for use in prophylactic or post-exposure treatment of infectious or respiratory diseases.

Abstract

An antiviral composition for use in the prevention or treatment of pathogenic microbial infections is provided. The liposomal antiviral composition of the composition comprises liposomes and an antiviral agent entrapped within the liposomes. The antiviral agent is stably encapsulated within the liposomes, and the resulting liposomal antiviral agent has been shown to be stably aerosolized or atomized for administration via the inhalation route, treating subjects in need with reduced side effects.

Description

[Technical Field]

[0001] The present invention relates to a drug delivery system for the delivery of antiviral agents. The present invention also relates to a method for preparing this drug delivery system. The present invention also relates to a sustained-release pharmaceutical composition suitable for a pulmonary delivery system with reduced systemic side effects. [Background technology]

[0002] Infectious diseases can be transmitted through different routes, including contact, droplet, and bloodborne transmission, and the bioavailability of drugs in the affected physical environment may be low for systemic drug administration. Penetration of drug delivery systems onto target cells in target tissues is a significant obstacle to the effective treatment of infectious diseases, such as lung infections, by inhalation. The retention of drug material within liposomes before the drug delivery system adheres to target epithelial cells may vary from liposomal drug to liposomal drug, based on the diffusion rate of free, uncharged drug material across the liposomal lipid membrane, which largely depends on the physicochemical properties of the lipid barrier in the presence of the microenvironment outside the liposome, as well as the aqueous environment inside the liposome.

[0003] Respiratory diseases caused by infection or other unknown reasons are extremely severe, debilitating lung diseases that lead to premature death, particularly those characterized by subsequent efficient viral replication and cell damage caused by virus-induced cytolysis or immunopathology. Infected cell lines and postmortem lung tissue showed cytopathological changes due to apoptosis, necrosis, or sometimes syncytial formation.

[0004] Liposomes have been used as drug carriers to mask the unpleasant taste of inhaled medications in the treatment of asthma, as described in U.S. Patent Application Publication No. 20110104259A1. Liposome encapsulation of drug substances may alter the pharmacokinetic profile of the drug substance, provide slower drug release in the local physical environment, enable optimal dosage with reduced drug administration frequency, and / or reduce side effects and toxicity. However, it is unknown whether quinine compounds or other antiviral agents delivered by whole liposomes via the inhalation route effectively exert the necessary functions, such as deposition on target cell lines expressing receptors suitable as docket sites for entry into intracellular target sites of the virus, and whether they achieve the desired pharmacokinetic profile in vivo.

[0005] It is not readily apparent that utilizing liposome technology to reconstitute antiviral agents could lead to inhaled liposomal formulations at prophylactic doses to prevent severe acute respiratory syndrome (ARDS) or therapeutic doses to treat respiratory or infectious diseases with reduced side effects. Currently, there are no practical liposomal drug formulations for inhalation as drug products for chemoprevention, such as prevention, treatment of mild cases, or treatment of severe acute respiratory syndrome (SARS) caused by infection with viruses such as coronavirus COVID-19, also known as SARS-CoV-2. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] U.S. Patent Application Publication No. 20110104259A1 [Overview of the project] [Problems that the invention aims to solve]

[0007] There remains an unmet need for inhalable formulations with predetermined encapsulation efficiency to achieve a balance by reducing the frequency and / or dosage of antiviral agents such as quinine and nucleoside compounds, and by targeting a desired prophylactic or therapeutic concentration range (window) for pulmonary delivery. In addition, formulations suitable for respiratory diseases should have characteristics such as being inhalable, exhibiting sufficient encapsulation efficiency after inhalation administration, having improved stability or appropriate resistance to destruction by topical substances such as pulmonary surfactants, and having a desired dose intensity to ensure the possibility of achieving the desired efficacy in the pulmonary environment. The present invention addresses this need and other needs. [Means for solving the problem]

[0008] The present invention provides a liposomal drug formulation, particularly for the treatment of respiratory or infectious diseases induced by inhalation, comprising at least one lipid, optionally phospholipids (multiple types may be used), sterols, and / or polyethylene glycol (PEG)-modified phospholipids, and an antiviral agent encapsulated within an aqueous liposome.

[0009] To improve existing treatment paradigms for respiratory or infectious diseases and to take advantage of the benefits of slow, sustained drug release, the inventors have developed an antiviral composition comprising a liposomal antiviral agent and a predetermined amount of free antiviral agent in an aqueous suspension that can be aerosolized and inhaled for the prophylactic or enhanced treatment of respiratory diseases. In particular, there is a need for an inhalable formulation for the prevention or treatment of SARS.

[0010] This disclosure provides compositions of antiviral agents for use in the prevention or treatment of respiratory or infectious diseases, particularly SARS, which have the following advantages: 1) achieving a longer therapeutic effect compared to inhaled free drug substances; 2) delivering the drug directly to the disease site or site of viral infection; 3) a faster onset of action; 4) reducing adverse drug reactions and systemic effects; 5) bypassing first-pass metabolism observed in oral administration and thus increasing the bioavailability of the drug substance (in addition, potentially reducing cardiotoxicity, ocular symptoms of retinopathy, nausea, vomiting, diarrhea and abdominal discomfort, gastrointestinal (GI) effects, and hepatotoxicity); 6) increasing the residence time of the drug substance in target tissues via sustained release from liposomal drugs; 7) reducing the frequency of drug administration; 8) non-invasive inhalation delivery; and / or 9) improving patient outcomes and compliance.

[0011] In certain embodiments, the antiviral agent according to the Disclosure is encapsulated in a predetermined amount within liposomes to achieve a composition with a preferred release profile and reduced toxicity, particularly cardiotoxicity, thereby forming the antiviral agent composition according to the Disclosure.

[0012] A composition of an antiviral agent for use in the treatment or prevention of respiratory disease, comprising an inhalable liposomal antiviral agent, wherein the liposomal antiviral agent is Liposomes containing at least one type of lipid, This liposome contains an antiviral agent and A composition of an antiviral agent containing the following is provided.

[0013] In some embodiments, the liposomes include a lipid bilayer composed of one or more phospholipids and sterols, where the sterol is cholesterol, and the phospholipid (multiple types are possible) to cholesterol ratio is 1:1 to 2:1, and optionally 3:2.

[0014] In some embodiments, the one or more phospholipids include phosphocholine (PC), which may be, but is not limited to, hydrogenated soy phosphatidylcholine (HSPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), or a mixture thereof. In some other embodiments, the one or more phospholipids include DSPC and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE) in a molar ratio of 1:1 or 3:2.

[0015] In some embodiments, the liposomal antiviral agent includes a 4-aminoquinoline compound.

[0016] In some embodiments, the 4-aminoquinoline compound is selected from the group consisting of chloroquine, hydroxy chloroquine, and amodiaquine.

[0017] [[ID=##]]In some embodiments, the antiviral agent includes a nucleoside compound of Structural Formula I,

Chemical Formula

[0018] In some embodiments, the composition of the antiviral agent according to the present disclosure further comprises an antibiotic, a supplement, an antiretroviral agent or a combination thereof. Examples of antibiotics are penicillins (ampicillin-sulbactam combination, piperacillin-tazobactam combination), macrolides, cephalosporins, aminoglycosides and glycopeptides. In some embodiments, the antibiotic is selected from the group consisting of curimycin and azithromycin.

[0019] In another aspect, the Disclosure relates to a composition of an antiviral agent for use in the prevention or treatment of an infectious or respiratory disease, or an aerosolized composition of particles containing such antiviral agent, wherein the concentration is at least 0.01 moles / mol, and optionally 0.01 moles / mol to 2.0 moles / mol, 0.05 moles / mol to 2.0 moles / mol, 0.05 moles / mol to 1.5 moles / mol, 0.05 moles / mol to 1.0 moles / mol, and 0.0 The present invention also provides compositions having a drug-to-lipid ratio of 5 mol / mol to 0.5 mol / mol, 0.05 mol / mol to 0.3 mol / mol, 0.05 mol / mol to 0.2 mol / mol, 0.05 mol / mol to 0.15 mol / mol, 0.01 mol / mol to 1 mol / mol, 0.05 mol / mol to 0.1 mol / mol, 0.07 mol / mol to 0.09 mol / mol, or about 0.085 mol / mol, as well as an antiviral agent concentration in the range of 0.1 mg / mL to 10 mg / mL.

[0020] In another aspect, the Disclosure also provides aerosolized particle compositions comprising a liposomal quinine compound for use in the prevention or treatment of respiratory diseases relating to the Disclosure, having a drug-to-lipid ratio of at least 0.01 mol / mol, optionally at least 0.05 mol / mol, optionally 0.01 mol / mol to 2.0 mol / mol, 0.05 mol / mol to 2.0 mol / mol, 0.05 mol / mol to 1.5 mol / mol, 0.05 mol / mol to 1.0 mol / mol, 0.05 mol / mol to 0.5 mol / mol, 0.05 mol / mol to 0.3 mol / mol, 0.05 mol / mol to 0.2 mol / mol, 0.05 mol / mol to 0.15 mol / mol, optionally about 0.5 mol / mol, and a concentration of the quinine compound in the range of 1 mg / mL to 10 mg / mL based on the composition.

[0021] In another aspect, the Disclosure also provides aerosolized particle compositions comprising a liposomal nucleoside compound for use in the prevention or treatment of infectious diseases relating to the Disclosure, having a drug-to-lipid ratio of at least 0.01 mol / mol, optionally at least 0.05 mol / mol, optionally 0.01 mol / mol to 1 mol / mol, 0.03 mol / mol to 0.5 mol / mol, 0.03 mol / mol to 0.15 mol / mol, 0.03 mol / mol to 0.1 mol / mol, optionally about 0.05 mol / mol, 0.05 mol / mol to 0.5 mol / mol, 0.05 mol / mol to 0.15 mol / mol, 0.05 mol / mol to 0.1 mol / mol, 0.07 mol / mol to 0.1 mol / mol, optionally about 0.085 mol / mol, and a concentration of the nucleoside compound of 0.1 mg / mL to 5 mg / mL based on the composition.

[0022] In another aspect, the Disclosure also provides a spray (inhalation spray) comprising a composition of an antiviral agent for use relating to the Disclosure.

[0023] In another aspect, the Disclosure also provides a particle aerosolized composition containing a composition of an antiviral agent for use in the prevention or treatment of respiratory or infectious diseases, the particle aerosolized composition containing the liposomal antiviral agent according to the Disclosure.

[0024] In another aspect, the Disclosure also provides a method for treating or preventing a respiratory or infectious disease, comprising the step of administering an effective amount of a composition of an antiviral agent for use in treating or preventing the respiratory disease relating to the Disclosure to a subject in need thereof.

[0025] In another embodiment, the Disclosure also provides a system for administering an antiviral composition to a subject in need thereof. The system comprises an antiviral composition according to the Disclosure and a lung delivery device. The lung delivery device can aerosolize the antiviral composition, and after aerosolization, the formed particles containing the liposomal antiviral agent contain an amount of free antiviral agent effective to provide immediate antiviral activity and an amount of the liposomal antiviral agent effective to provide sustained antiviral activity.

[0026] In another aspect, the Disclosure also provides a method for mitigating complications associated with the treatment of respiratory or infectious diseases in human subjects, the method comprising administering a composition according to the Disclosure to a subject in need thereof. According to the Disclosure, complications include, but are not limited to, cardiotoxicity or hepatotoxicity. According to the Disclosure, complications include, but are not limited to, prolongation of the corrected QT time (QTc).

[0027] Other subjects, advantages, and novel features of this disclosure will become more apparent from the following detailed description, in conjunction with the accompanying drawings. [Brief explanation of the drawing]

[0028] [Figure 1] Figure 1 shows the pharmacokinetic profile of HCQ in rat lungs after administration of the composition and free HCQ according to this disclosure. [Figure 2] Figure 2 shows the pharmacokinetic profile of HCQ in rat blood after administration of the composition and free HCQ according to this disclosure. [Figure 3] Figure 3 shows the pharmacokinetic profile of HCQ in rat hearts after administration of the composition and free HCQ according to this disclosure. [Figure 4A]Figures 4A and 4B show a series of graphs illustrating the mean concentration-time profiles of GS-441524 in the lungs (Figure 4A) and plasma (Figure 4B) of rats after a single IV administration of SBECD formulation GS-441524 (GS-441524 Solution-IV) or a single IT administration of liposomal GS-441524, also represented as ISPM21 (ISPM21-IT). LLOQ: Limit of Quantification. [Figure 4B] Figures 4A and 4B show a series of graphs illustrating the mean concentration-time profiles of GS-441524 in the lungs (Figure 4A) and plasma (Figure 4B) of rats after a single IV administration of SBECD formulation GS-441524 (GS-441524 Solution-IV) or a single IT administration of liposomal GS-441524, also represented as ISPM21 (ISPM21-IT). LLOQ: Limit of Quantification. [Modes for carrying out the invention]

[0029] As used above and throughout this disclosure, the following terms shall be understood to have the following meanings unless otherwise specified.

[0030] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly contradicts them.

[0031] All numerical values ​​in this specification may be understood to be modified by “approximately,” meaning that when referring to measurable values ​​such as quantity or duration, “approximately” includes a variation of ±10%, preferably ±5%, more preferably ±1%, and even more preferably ±0.1% from the specified value. This is because, unless otherwise stated, such variation is appropriate for obtaining the desired amount of liposomal drug.

[0032] As used herein, the terms “to treat,” “to treat,” “to be treated,” “to be treated,” “treatment,” or “treatment” include preventive (e.g., prophylactic), mitigating, and curative uses or results. The term “subject” includes vertebrates having a respiratory disease or other disease or suspected viral infection. Preferably, the subject is a warm-blooded animal, preferably a mammal, preferably a human.

[0033] As used herein, the term “drug” refers to an antiviral agent, such as a quinine compound or nucleoside compound, related to its activity for the desired therapeutic effect relating to this disclosure. As used herein, the term “drug-to-lipid ratio (D / L)” refers to the ratio of an antiviral agent to at least one lipid in the composition relating to this disclosure. The drug content in the free drug or liposomal drug of the liposomal drug composition relating to this disclosure can be determined by ultraviolet-visible absorbance measurement or high-performance liquid chromatography (HPLC), but is not limited to these methods. The phospholipid content or concentration of liposomes and liposomal drugs can be determined by assaying the phosphorus content of liposomes and liposomal drug samples using a phosphorus assay (adapted from G. Rower et al., Lipids 1970, 5, 494-496) or HPLC.

[0034] As used herein, pharmacokinetic data were obtained in rats; however, pharmacokinetic profiles correlated with the inhalation compositions relating to this disclosure can be obtained in other mammals, including but not limited to cats, dogs, horses, mice, pigs, non-human primates, and humans, for the purpose of developing the inhalation compositions.

[0035] Infectious diseases and respiratory diseases In this disclosure, infectious diseases and pathogenic infections refer to disorders caused by living organisms such as viruses, parasites, and bacteria. In one embodiment, an infectious disease is transmitted through fecal-oral transmission, droplet contact, sexual transmission, oral transmission, direct contact, vector-borne transmission, vertical transmission (mother-to-child transmission), iatrogenic transmission, or host-borne transmission. In another embodiment, an infectious disease may include, but is not limited to, urinary tract infections, skin infections, respiratory infections, odontogenic infections, vaginal infections, and intraamniotic infections.

[0036] In some embodiments, infectious diseases include acute flaccid osteomyelitis (AFM), anaplasmosis, anthrax, babesiosis, botulism, brucellosis, campylobacterium infection, carbapenem-resistant infection (CRE / CRPA), chancroid, chikungunya virus infection (chikungunya), chlamydia, siguatella (harmful algal bloom (HABS)), Clostridium difficile infection, Clostridium perfringens (epsilon toxin), coccidioidomyces (Coccidioidomyces) fungal infection (valley fever), COVID-19 (novel coronavirus infection), Creutzfeldt-Jakob disease, Infectious spongiform encephalopathy (CJD), cryptosporidiosis (crypto), cyclosporiasis, dengue fever, 1, 2, 3, 4 (dengue fever), diphtheria, Escherichia coli infection, Shiga toxin production (STEC), Eastern equine encephalitis (EEE), Ebola hemorrhagic fever (Ebola), ehrlichiosis, encephalitis, arboviral or post-infectious, enterovirus infection, non-polio (non-polioenterovirus), enterovirus infection D68 (EV-D68), giardiasis (giardia), glanders, gonorrhea, inguinal granuloma, Haemophilus influenzae infection, type b (HIB or h-flu), hantavirus pulmonary syndrome (HPS), hemolytic uremic syndrome (HUS), hepatitis A (Hep A), Hepatitis B (Hep B), Hepatitis C (Hep C), Hepatitis D (Hep D), Hepatitis E (HepE) Herpes, Zoster, VZV (herpes zoster), histoplasmosis infection, human immunodeficiency virus / AIDS (HIV / AIDS), human papillomavirus (HPV), influenza (flu), lead poisoning, Legionnaires' disease, leprosy (Hansen's disease), leptospirosis, listeriosis (listeria), Lyme disease, lymphogranulomatosis of the inguinal region (LGV), malari A. Measles, meridianus, meningitis, viral (meningitis, viral), meningococcal disease, bacterial (meningitis, bacterial), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), Multiple System Inflammatory Syndrome in Children (MIS-C), mumps, norovirus, paralytic shellfish poisoning (paralytic shellfish poisoning) (paralytic shellfish poisoning, ciguatera), lice infestation (lice, head and body lice), pelvic inflammatory disease (PID), pertussis (whooping cough) cough), plague (bubonic plague, septic plague, pneumonic plague), pneumococcal disease (pneumonia), polio, poissans, psittacosis (parrot fever), lice infestation (pubic lice; groin lice infestation), pustular rash diseases (smallpox, monkeypox, cowpox), Q fever, rabies, ricin poisoning, rickettsial diseases (Rocky Mountain spotted fever), rubella (including congenital rubella (germanMeasles), Salmonellosis (Salmonella), Sarcoptes scabiei infestation (Sarcoptes scabiei), Sconbroid (Sconbroid food poisoning), Septic shock (Sepsis), Severe acute respiratory syndrome (SARS), Bacterial gastroenteritis (Shigella), Smallpox, Staphylococcal infection, Methicillin resistance (MRSA), Staphylococcal food poisoning, Enterotoxin B poisoning (Staphylococcal food poisoning), Staphylococcal infection, Vancomycin moderate resistance (VISA), Staphylococcal infection, Vancomycin resistance (VRSA), Streptococcal disease, Group A (invasive) (Group A Streptococcus (invasive)), Streptococcal disease, Group B (Group B Streptococcus), Streptococcal toxin shock syndrome, STSS, Toxin shock (STSS, TSS), Syphilis, Stage 1, Stage Examples of diseases that fall under this category include, but are not limited to, stage 2, early latent, late latent, congenital, tetanus infection, tetanus (bite spasm), trichomoniasis (trichomoniasis), trichinellosis (trichinosis), tuberculosis (TB), tuberculosis (latent) (LTBI), tularemia (rabbit fever), typhoid fever (group D), typhoid fever, vaginosis, bacterial infection (yeast infection), vaping-related lung injury (e-cigarette-related lung injury), varicella (chickenpox), Vibrio cholerae (cholera), vibriosis (vibrio), viral hemorrhagic fever (Ebola, LASSA, Marburg), West Nile virus infection, yellow fever, Yersenia (Yersinia), and Zika virus infection (Zika).

[0037] Respiratory diseases relating to this disclosure include, but are not limited to, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or severe acute respiratory syndrome (SARS), which are accompanied by major complications including fluid leakage into the lungs, making breathing difficult or impossible. Typical symptoms include fever, cough, wet cough, dry cough, dyspnea and fatigue or muscle pain, chest tightness, and a gradual onset of shortness of breath. Complications include pulmonary hypertension, heart failure, pneumonia, or pulmonary embolism.

[0038] In some embodiments, the antiviral compositions according to this disclosure are suitable for use in the prevention, treatment of mild cases, and therapy of acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or severe acute respiratory syndrome (SARS) caused by coronavirus or its derivatives.

[0039] Liposomes and liposomal antiviral agents As used herein, the terms “liposome” or “liposomal” refer to a group of vesicles characterized in that each vesicle has an aqueous internal space isolated from an outer medium by one or more bilayer membranes (bilayers). The bilayer membrane of a liposome is typically formed by one or more lipids, i.e., synthetic or naturally occurring amphiphilic molecules containing spatially separated hydrophobic and hydrophilic domains.

[0040] The internal aqueous space of the liposome is substantially free from neutral lipids such as triglycerides, non-aqueous phases (oil phases), water-oil emulsions, second liposomes, or other mixtures containing non-aqueous phases. Non-limiting examples of liposomes include small unilamellar vesicles (SUVs) and large unilamellar vesicles (LUVs), as well as multilayer vesicles (MLVs), having average diameters in the ranges of 50nm-10000nm, 50nm-500nm, 50nm-450nm, 50nm-400nm, 50nm-350nm, 50nm-300nm, 50nm-250nm, 50nm-200nm, 100nm-500nm, 100nm-450nm, 100nm-400nm, 100nm-350nm, 100nm-300nm, 100nm-250nm, or 100nm-200nm, which can pass through sterile filters. For example, MLV may be formed directly from a hydrated lipid thin film, spray-dried powder, or freeze-dried cake of a selected lipid composition having a scavenging agent, and SUV and LUV can be resized from MLV by sonication, homogenization, microfluidization (micro-solution operation), or extrusion.

[0041] Generally, liposomes typically contain a lipid mixture comprising at least one lipid selected from the group consisting of dialiphatic chain lipids, such as phospholipids, diglycerides, dialiphatic glycolipids, monolipids, such as sphingomyelin and sphingoglycolipids, sterols, such as cholesterol, and derivatives thereof and combinations thereof.

[0042] Examples of phospholipids relating to this disclosure include 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimiristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine (PSPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), and 1,2-distearoyl-sn-glycero-3-phosphocholine. Hocoline (DSPC), 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), Hydrogenated soybean phosphatidylcholine (HSPC), 1,2-Dimyristoyl-sn-glycero-3-phospho-(1'-rac-glycerol)(sodium salt) (DMPG), 1,2-Dipalmitoyl-sn-glycero-3-phospho-(1'-rac-glycerol)(sodium salt) (DPPG), 1-Palmitoyl-2-stearoyl-sn-glycero-3-phospho-(1'-rac-glycerol)(sodium salt) 1,2-Distearoyl-sn-glycero-3-phospho-(1'-rac-glycerol)(sodium salt)(DSPG), 1,2-Dioleoyl-sn-glycero-3-phospho-(1'-rac-glycerol)(DOPG), 1,2-Dimyristoyl-sn-glycero-3-phospho-L-serine(sodium salt)(DMPS), 1,2-Dipalmitoyl-sn-glycero-3-phospho-L-serine(sodium salt)(DPPS), 1,2-Distearoyl-sn-glycero-3-phospho -L-serine (sodium salt) (DSPS), 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt) (DMPA), 1,2-dipalmitoyl-sn-glycero-3-phosphate (sodium salt) (DPPA), 1,2-distearoyl-sn-glycero-3-phosphate (sodium salt) (DSPA), 1,2-dioleoyl-sn-glycero-3-phosphate (sodium salt) (DOPA), 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-Dipalmitoyl-sn-glycero-3-phospho(1'-myoyl Examples include, but are not limited to, L-α-phosphatidylethanolamine (EPE), L-α-phosphatidylethanolamine (EPE), L-α-phosphatidylethanolamine (EPE). Other examples include 1,2-distearoyl-sn-glycero-3-phosphoinositol (ammonium salt) (DSPI), 1,2-dioleoyl-sn-glycero-3-phospho-(1'-myoinositol) (ammonium salt) (DOPI), cardiolipin, L-α-phosphatidylcholine (EPC), and L-α-phosphatidylethanolamine (EPE).

[0043] The liposomal antiviral agents relating to this disclosure optionally incorporate polyethylene glycol (PEG)-modified phosphatidylethanolamine (PE) into the vesicle membrane, thereby incorporating a meaningful amount of the PEG portion into the vesicle surface, achieving safe, effective, low-dosage, longer-lasting, and sustained drug release.

[0044] The liposomal antiviral agent relating to this disclosure optionally incorporates PEG-modified phosphatidylethanolamine (PE) or a fatty acid into the vesicle membrane, thereby incorporating a significant amount of negatively charged material onto the vesicle surface and preventing liposome aggregation or coagulation (cotton-like precipitate) processes in storage solution.

[0045] The polyethylene glycol-modified lipids described above include a polyethylene glycol moiety conjugated with a lipid. In some embodiments, the PEG moiety has a molecular weight of about 5,000 to about 20,000 daltons. In certain embodiments, the PEG-modified lipids are mixed with phospholipids to form liposomes having one or more bilayer membranes. In some embodiments, the amount of PEG-modified lipids is in the range of 0.0001 mol% to 40 mol%, optionally 0.001 mol% to 30 mol%, optionally 0.01 mol% to 20 mol%, optionally 0.0001 mol% to 10 mol%, optionally 0.001 mol% to 5 mol%, particularly 6 mol% or less, optionally 5 mol% or less, 3 mol% or less, or 2 mol% or less, based on total phospholipids and sterols. In some embodiments, the PEG-modified lipids have a PEG moiety having an average molecular weight in the range of 1,000 g / mol to 5,000 g / mol. In certain embodiments, the PEG-modified lipid is phosphatidylethanolamine linked to a polyethylene glycol group (PE-PEG). In further embodiments, the PEG-modified phosphatidylethanolamine is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)](DSPE-PEG).

[0046] In certain embodiments, the PEG-modified phosphatidylethanolamine (PE) is DSPE-PEG in an amount ranging from 0.0001 mol% to 40 mol%, optionally from 0.01 mol% to 20 mol%, of the total lipid content of the liposome, and has a PEG moiety with an average molecular weight of 2,000 g / mol.

[0047] The terms “liposomal antiviral agent” and “liposomal drug” are used interchangeably in this disclosure. The liposomal antiviral agents relating to this disclosure include antiviral-captured liposomes, which are prepared by encapsulating the antiviral agent within the aqueous interior of liposomes using a transmembrane pH gradient-driven remote loading method.

[0048] In some embodiments, liposomes are formed together with a drug substance such as hydroxychloroquine or GS-441524 to encapsulate the drug substance within the aqueous interior of the liposome, or are formed alone as empty liposomes with a transmembrane gradient for later use in a drug loading process, such as an active loading method also known as remote loading, for forming liposomal drugs.

[0049] In some embodiments, a transmembrane pH gradient is created by using a scavenger for remotely loading antiviral agents into liposomes, the scavenger consisting of ammonium compounds and anionic counterions.

[0050] The term "ammonium compound" is NR4 + The compound comprises an unsubstituted or substituted ammonium which is a cationic ion presented by the formula, where each R is independently H or an organic residue, which is independently alkyl, alkylidene, heterocyclic alkyl, cycloalkyl, aryl, alkenyl, cycloalkenyl, or a hydroxyl-substituted derivative thereof, which optionally contains an S, O, or N atom in its hydrocarbon chain to form an ether, ester, thioether, amine, or amide bond. In one embodiment, the ammonium compound is ammonium.

[0051] The term "anionic counterion" refers to an anionic ion, or an entity covalently bonded to one anionic functional group. Anionic ions or anionic functional groups have a negative charge under physiological conditions.

[0052] The anionic ion or anionic functional group can be selected from one or more of the following: sulfate anion, citrate anion, sulfonate anion, phosphate anion, pyrophosphate anion, tartrate anion, succinate anion, maleate anion, borate anion, carboxylate anion, bicarbonate anion, glucuronate anion, chloride anion, hydroxide anion, nitrate anion, cyanate anion, or bromide anion.

[0053] In one embodiment, the anionic ion and the anionic functional group are selected from one or more of the following: citrate anion, sulfate anion, sulfonate anion, phosphate anion, pyrophosphate anion, and carboxylate anion.

[0054] In yet another embodiment, the entity linked to the anionic functional group may be a natural or synthetic organic or inorganic compound. Examples of such entities include, but are not limited to, alkyl or aryl groups, and nonpolymeric substances selected from, for example, benzene, nucleotides, and sugars. Alkyl refers to a saturated hydrocarbon radical having a specified number of carbon atoms. For example, alkyl refers to alkyl(C) with 1 to 4 carbon atoms. 1~4 Alkyl) and alkyl (C) with 1 to 6 carbon atoms. 1~6 Alkyl), alkyl (C) with 1 to 8 carbon atoms 1~8 Alkyl) and alkyl (C) with 1 to 10 carbon atoms. 1~10 Alkyl) and alkyl (C) with 1 to 12 carbon atoms. 1~12 Alkyl) and alkyl (C) with 1 to 14 carbon atoms. 1~14 Alkyl) and alkyl (C) with 1 to 16 carbon atoms. 1~16 Alkyl) and alkyl (C) with 1 to 18 carbon atoms. 1~18 Alkyl) and alkyl(C) with 1 to 20 carbon atoms 1~20 Selected from the group consisting of alkyl groups.

[0055] In some embodiments, the anionic counterion is selected from the group consisting of sulfate anions, phosphate anions, citrate anions, gluconate anions, octasulfate sucrose anions, dextran sulfate anions, and combinations thereof.

[0056] In some embodiments, the scavenger is selected from the group consisting of ammonium sulfate, ammonium phosphate, ammonium citrate, sucrose ammonium octasulfate, ammonium dextran sulfate, dimethylammonium sulfate, dimethylammonium phosphate, dimethylammonium citrate, diethylammonium sulfate, diethylammonium phosphate, diethylammonium citrate, sucrose diethylammonium octasulfate, diethylammonium dextran sulfate, trimethylammonium sulfate, trimethylammonium phosphate, trimethylammonium citrate, triethylammonium sulfate, triethylammonium phosphate, triethylammonium citrate, sucrose triethylammonium octasulfate, triethylammonium dextran sulfate, copper gluconate, copper glucuronate, and combinations thereof.

[0057] In some embodiments, the liposomal antiviral agent has an average particle size of 50 nm to 1,000 nm. Non-limiting examples of liposomal antiviral agents have an average diameter in the range of 50 nm to 20 μm, 50 nm to 10 μm, 50 nm to 1,000 nm, 50 nm to 500 nm, 50 nm to 400 nm, 50 nm to 300 nm, 50 nm to 250 nm, 50 nm to 200 nm, 100 nm to 300 nm, or 150 nm to 250 nm.

[0058] In some embodiments, the antiviral agent may include, but is not limited to, an antimalarial agent, an antiretroviral agent, or a combination thereof. In particular, the antiviral agent is selected from the group consisting of quinine compounds, nucleoside compounds, and combinations thereof.

[0059] The term "quinine compound" refers to substances derived from quinine, a lead compound with antimalarial activity extracted from the bark of trees in the Cinchona genus. Quinine compounds, such as hydroxychloroquine, have been shown to have potential in inhibiting pneumonia exacerbations, improving imaging findings, promoting viral negativity, and shortening disease progression. However, systemic administration of quinine compounds may cause side effects such as blurred vision, nausea, vomiting, abdominal colic, headache, and diarrhea.

[0060] The quinine compounds relating to this disclosure include, but are not limited to, quinine and other 4-aminoquinolines, such as quinine, quinidine, cinconine, cinconidine, chloroquine (CQ), and hydroxychloroquine (HCQ). Exemplary quinine compounds, CQ and HCQ, have been suggested to prevent acidification of intracellular organelles and inhibit the lysosomal release of the viral genome. In addition, these drugs can interfere with the glycosylation of the angiotensin-converting enzyme-2 (ACE2) receptor on host cells, thereby reducing the binding efficiency between the receptor and the spike protein on the surface of the coronavirus.

[0061] Nucleosides have been reported as inhibitors of non-structural viral proteins, such as RNA-dependent RNA polymerase, and are being used as a promising treatment for RNA virus infections. Nucleosides are expected to be taken up by cells and converted to triphosphates in vivo, competing with polymerase nucleotide binding sites and terminating the polymerase chain reaction. This conversion to triphosphates is usually mediated by cell kinases, which imposes further structural requirements on potential nucleoside polymerase inhibitors. The formation of monophosphate by nucleoside kinases is generally considered the rate-limiting step of the three phosphorylation events. U.S. Patent No. 7,964,580 discloses a pronucleoside containing a phosphoramidate moiety masked with a neutral lipophilic group to obtain a suitable partition coefficient for optimizing cellular uptake and transport, dramatically increasing the intracellular concentration of nucleoside monophosphate compared to administration of the parent nucleoside alone. Although there are controversial observational predictions, the enzyme-mediated hydrolysis of the phosphate ester moiety likely generates the nucleoside itself, rather than the target nucleoside monophosphate, immediately after circulation, before its targeting to the desired site. For example, when administered intravenously, the known nucleoside compound (2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazine-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-carbonitrile (also known as GS-441524) is a more stable metabolite of remdesivir than its monophosphate form (Humeniuk R, Mathias A, Cao H et al., Safety, Tolerability, and Pharmacokinetics of Remdesivir, An Antiviral for Treatment of COVID-19, in Healthy Subjects. Clin Transl Sci. 2020;13(5):896-906. doi:10.1111 / cts.12840).The efficient delivery of nucleoside compounds to enriched target cell sites with respect to critical rate-limiting nucleoside kinases may be a universal platform (solution) for avoiding the complex manufacturing processes of different pronucleosides for a wide range of nucleoside compounds.

[0062] In some embodiments, the antiviral agent compositions relating to this disclosure further include antibiotics, supplements, or combinations thereof.

[0063] In some embodiments, the antiviral agent comprises one or more 1'-substituted carbanucleoside compounds or 2'-substituted carbanucleoside compounds as described in U.S. Patent No. 8,008,264 and U.S. Patent No. 9,481,704.

[0064] In some embodiments, the antiviral agent is directed to nucleoside compounds, including but not limited to 1'-substituted carbanucleoside compounds and their pharmaceutically acceptable salts.

[0065] In some embodiments, the antiviral agent comprises an RNA-dependent RNA virus polymerase inhibitor, which comprises a nucleoside compound of structural formula (I). [ka] Each R 1 , R 2 , R 3 , R 4 or R 5 H, OR a , N(R a )2, N3, CN, NO2, S(O) n R a , halogen or methyl, and n is 0, 1 or 2, R 6 is CN or H, Each R aThese are independently H, (C1~C8) alkyl, (C2~C8) alkenyl, (C2~C8) alkynyl, aryl (C1~C8) alkyl, (C4~C8) carbocykylalkyl, and -C(=O)R 11 , -C(=O)OR 11 -C(=O)NR 11 R 12 -C(=O)SR 11 ,-S(O)R 11 -S(O)2R 11 -S(O)(OR 11 ), -S(O)2(OR 11 ), or -SO2NR 11 R 12 And, R 7 H is H, each X 1 or X 2 CR 10 or N, R 8 Halogen, NR 11 R 12 , N(R 11 )(OR 11 ), NR 11 NR 11 R 12 , N3, NO, NO2, CHO, CN, -CH(=NR 11 ), -CH=NHNR 11 -CH=N(OR 11 ), -CH(OR 11 )2, -C(=O)NR 11 R 12 -C(=S)NR 11 R 12 , -C(=O)OR 11 (C1~C8)alkyl, (C2~C8)alkenyl, (C2~C8)alkynyl, aryl(C1~C8)alkyl, (C4~C8)carbocykylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)(C1~C8)alkyl, -S(O) n (C1-C8) alkyl, aryl (C1-C8) alkyl, OR 11 or SR 11 Each aryl or heteroaryl may be independently substituted with one or more Z groups. Each R 9 or R 10 These are H, halogen, and R independently. 11 , OR 11 , SR 11 , NR 11 R 12 , N(R 11 )(OR 11 ), NR 11 NR 11 R 12 , N3, NO, NO2, CHO, CN, -CH(=NR 11 ), -CH=NHNR 11 -CH=N(OR 11 ), -CH(OR 11 )2, -C(=O)NR 11 R 12 -C(=S)NR 11 R 12 , -C(=O)OR 11 And, Each R 11 Or R 12 These are independently H, (C1~C8)alkyl, (C2~C8)alkenyl, (C2~C8)alkynyl, (C4~C8)carbocykylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)(C1~C8)alkyl, -S(O) n (C1-C8)alkyl or aryl(C1-C8)alkyl, where each aryl or heteroaryl may be independently substituted with one or more Z groups. Or, R 11 Or R 12 These atoms, along with the nitrogen atoms to which they are bonded, form a 3- to 7-membered heterocycle, and any one carbon atom in this heterocycle can be optionally -O-, -S-, or -NR. a - may be replaced with Each Z group independently produces a halogen, -O - ,=O,-OR b , -SR b , -S - , -NR b 2, -N + R b 3, =NR b, -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO2, =N2, -N3, -NHC(=O)R b -OC(=O)R b , -NHC(=O)NR b 2, -S(=O)2-, -S(=O)2OH, -S(=O)2R b -OS(=O)2Or b -S(=O)2OH, -S(=O)R b , -OP(=O)(OR b )2, -P(=O)(OR b )2, -P(=O)(O - )2, -P(O)(OR b )(O - ), -C(=O)R b -C(=O)X, -C(S)R b , -C(O)OR b ,-C(O)O - , -C(S)OR b -C(O)SR b ,-C(S)SR b -C(O)NR b 2, -C(S)NR b 2, -C(=NR b )NR b 2, and each R b Each is independently H, alkyl, aryl, arylalkyl, or heterocyclic ring, and one or more of the non-terminal carbon atoms of each of the above (C1-C8) alkyls are -O-, -S-, or -NR a It may be replaced with -.

[0066] In some embodiments, the antiviral agent is [ka] [ka] Alternatively, a selection is made from the group consisting of these pharmaceutically acceptable salts.

[0067] In one embodiment, the liposomal antiviral agent comprises a lipid bilayer containing one or more phospholipids, sterols, and optionally selected polyethylene glycol (PEG)-modified lipids, particularly PEG-modified phosphatidylethanolamine (PEG-PE), and an aqueous interior enclosed by this lipid bilayer and containing one or more antiviral agents.

[0068] In one embodiment, the one or more phospholipids are neutral phospholipids, and the PEG-modified lipid is DSPE-PEG. The amount of DSPE-PEG is in the range of 0.001 to 5 mol%, optionally 0.0001 to 40 mol%, optionally less than 6 mol%, and optionally 0.001 to 30 mol%, based on total phospholipids and sterols.

[0069] In one embodiment, the liposomal antiviral agent composition has a drug-to-lipid ratio (ratio of antiviral agent to at least one lipid) of at least 0.01 mol / mol to 0.1 mol / mol, and comprises a lipid bilayer containing DPPC and cholesterol, and an aqueous interior containing the antiviral agent enclosed by the lipid bilayer and captured by a scavenger, wherein the antiviral agent is (2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazine-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-carbonitrile (GS-441524), and the scavenger is ammonium sulfate.

[0070] Inhalable compositions and their aerosolized particles The antiviral compositions relating to this disclosure are adapted for the preparation of inhalable aerosolized compositions of particles containing the liposomal antiviral agents described above. These compositions can be administered by inhalation as a spray or aerosol, or by intrathecal administration. Inhalation administration is preferred. The overall results are lower administration frequency and higher therapeutic index compared to free drugs or parenteral forms thereof. The liposomal antiviral agents in these compositions are particularly advantageous due to their ability to protect the drug while being compatible with the pulmonary lining or pulmonary surfactants.

[0071] In one embodiment, the antiviral agent composition according to the Disclosure has a drug-to-lipid ratio (D / L) in the range of at least 0.01 mol / mol, optionally at least 0.1 mol / mol, preferably 0.05 mol / mol to 1 mol / mol, optionally 0.1 mol / mol to 0.7 mol / mol, optionally 0.15 mol / mol to 0.6 mol / mol, and optionally 0.15 mol / mol to 0.2 mol / mol. The drug-to-lipid ratio refers to the molar ratio of the antiviral agent to at least one lipid. In certain embodiments, the at least one lipid comprises a neutral phospholipid and a sterol in a molar ratio of 1:1 or 3:2. Optionally, the neutral phospholipid is DPPC and the sterol is cholesterol.

[0072] In one embodiment, the antiviral agent composition contains at least one lipid in concentrations ranging from 1 mM to 200 mM, 1 mM to 100 mM, 5 mM to 100 mM, 10 mM to 180 mM, 15 mM to 140 mM, 20 mM to 160 mM, 30 mM to 140 mM, and 40 mM to 120 mM. Alternatively, the antiviral agent composition contains one or more phospholipids in concentrations ranging from 1 mM to 100 mM, 5 mM to 100 mM, 5 mM to 90 mM, 10 mM to 80 mM, 15 mM to 70 mM, or 20 mM to 60 mM.

[0073] In one embodiment, the antiviral composition has a total concentration of the quinine compound in the range of 0.1 mg / mL to 80 mg / mL, 0.5 mg / mL to 60 mg / mL, 1 to 30 mg / mL and 2 mg / mL to 15 mg / mL, 0.5 mg / mL to 70 mg / mL, 0.5 mg / mL to 60 mg / mL, 0.5 mg / mL to 50 mg / mL, 0.5 mg / mL to 40 mg / mL, 0.5 mg / mL to 30 mg / mL, 0.5 mg / mL to 20 mg / mL, 0.5 mg / mL to 10 mg / mL, 0.5 mg / mL to 8 mg / mL, 0.5 mg / mL to 5 mg / mL, 1.0 mg / mL to 6 mg / mL, 1.5 mg / mL to 5.0 mg / mL, 1.5 mg / mL to 4.0 mg / mL, or about 2.0 mg / mL.

[0074] In one embodiment, the antiviral agent composition has one or more phospholipids at concentrations in the range of 1 mM to 100 mM, 5 mM to 100 mM, 5 mM to 90 mM, 10 mM to 80 mM, 15 mM to 70 mM, or 20 mM to 60 mM, and a drug-to-lipid (D / L) ratio in the range of 0.01 mol / mol to 1 mol / mol, 0.03 mol / mol to 0.5 mol / mol, 0.03 mol / mol to 0.15 mol / mol, 0.03 mol / mol to 0.1 mol / mol, about 0.005 mol / mol, 0.05 mol / mol to 0.1 mol / mol, 0.07 mol / mol to 0.09 mol / mol, or 0.085 mol / mol, and the antiviral agent is a 1'-substituted carbanucleoside compound or a 2'-substituted carbanucleoside compound having a free 5'-OH group.

[0075] In some embodiments, the liposomal antiviral agent composition further comprises a free antiviral agent, the free antiviral agent of the composition according to the disclosure being in an amount of less than 60%, less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, or 10% to 40%, 15% to 35%, or 10% to 30% of the total amount of antiviral agent of the composition.

[0076] In some embodiments, aerosolized compositions of particles containing the compositions of the present disclosure are produced by aerosolizing the compositions using a nebulizer (nebulizer, atomizer) selected from the group consisting of an air jet nebulizer, an ultrasonic nebulizer, a vibrating mesh nebulizer, a condensing aerosol generator, an electrohydrodynamic nebulizer, or other lung delivery devices known in the art.

[0077] In some embodiments, the aerosolized composition of the particles has an aerodynamic mass median particle diameter of 0.5 μm to 5 μm, and optionally 1 μm to 3 μm.

[0078] Following aerosolization, leakage of the antiviral agent from the liposomes of the liposomal antiviral agent in the composition according to this disclosure results in a portion of the antiviral agent becoming free and not captured by the liposomes. The resulting free antiviral agent in the aerosolized composition is less than 60%, less than 50%, less than 40%, less than 30%, less than 20%, less than 10%, or less than 5%, and is optionally a controlled proportion within the ranges of 0.1% to 50%, 0.5% to 40%, 0.5% to 30%, 0.5% to 20%, 0.5% to 10%, 0.5% to 5%, 10% to 50%, 15% to 45%, 20% to 45%, and 25% to 35%.

[0079] In specific embodiments, the aerosolized composition of the particles is subjected to pulmonary delivery to a subject requiring a release rate of approximately 0.5% to 25% per hour of the administered drug dose, with complete release of the antiviral agent occurring at least approximately 12 to 24 hours later.

[0080] This disclosure will be further described with reference to the following specific, non-limiting embodiments. [Examples]

[0081] The following examples illustrate the preparation and characteristics of specific embodiments of the present disclosure.

[0082] Example 1: Stability of liposomal antiviral agents The liposomal antiviral agents relating to this disclosure include liposomes containing the antiviral agent, which are prepared by active or passive loading known in the art.

[0083] A. Preparation of liposomal antiviral agents by active loading I. Preparation of empty liposomes The process for preparing empty liposomes for remote loading was carried out by thin-film hydration or solvent injection. These methods may include the following steps: 1. Weigh a predetermined molar ratio of phospholipids and cholesterol lipid mixture in the presence or absence of DSPE-PEG2000, and add them to 10 mL of chloroform in a round-bottom flask. 2. The process involves attaching a flask to a rotary evaporator at an appropriate temperature according to the lipid composition, stirring the flask to dissolve the lipid mixture, and then, while stirring the flask, evaporating the chloroform under vacuum to obtain a dry lipid thin film. 3. A step of preparing a scavenger solution (e.g., ammonium sulfate (AS)) by adding a scavenger to distilled water, vortexing the solution to dissolve the powder, 4. Add the above scavenging agent solution to the dried lipid thin film and stir at an appropriate temperature according to the lipid composition to form a liposome solution. 5. This liposome solution is frozen and thawed using liquid nitrogen and a water bath at an appropriate temperature depending on the lipid composition to obtain a liposome sample. 6. The obtained liposome sample is extruded through a 0.2 μm polycarbonate membrane and a 0.1 μm polycarbonate membrane at an appropriate temperature according to the lipid composition to obtain the designed particle size. 7. Dialysis of the extruded liposome sample to remove free scavenging agents, followed by adding the sample to a dialysis bag (MWCO: 25kD), sealing the bag, agitating the dialysis bag in 100x volume of 9.4% (w / v) sucrose solution, physiological saline, or appropriate buffer, and further replacing the sucrose solution, physiological saline, or appropriate buffer after 1 hour and 4 hours, and agitating overnight. 8. A step to obtain empty liposomes by sterilizing the dialyzed liposome sample by filtering it through a 0.45 μm PTFE membrane.

[0084] II. Drug loading of antiviral agents into liposomes to obtain liposomal antiviral agents The following method represents a typical protocol for encapsulation of hydroxychloroquine or chloroquine into liposomes by remote loading, and this protocol includes the following steps. 1. A step of preparing a solution of hydroxychloroquine or chloroquine at a concentration of 40 mg / mL or an appropriate concentration in 9.4% (w / v) sucrose or an appropriate medium, and heating it for a short time at an appropriate temperature to obtain a stock solution containing hydroxychloroquine or chloroquine (hereinafter referred to as "stock solution"). 2. A step of mixing empty liposomes, physiological saline, and stock solution prepared by the process according to Section A(I) of Example 1 [in a typical embodiment, conditions of a DPPC:cholesterol molar ratio of 3:2, 300 mM ammonium sulfate (AS), and 30 mM phospholipid concentration are used] together in a conical tube to obtain a loading solution with a target D / L ratio of 100 g / mol or 0.19 mol / mol. 3. The loading solution is continuously shaken at an appropriate temperature for 30 minutes or the designed time to form a sample of drug-loaded liposomes. 4. If necessary, a step of removing free drugs, changing the buffer solution, or adjusting the drug concentration by dialysis or membrane-based tangential flow filtration (TFF), and 5. The drug encapsulation amount (i.e., loading efficiency) of the final sample is determined using size exclusion column chromatography and HPLC analysis to obtain liposomal antiviral compositions with drug concentrations in the range of 2 mg / mL to 10 mg / mL and an antiviral agent-to-lipid ratio of 0.05 mol / mol to 1.5 mol / mol, based on the entire composition (see formulation numbers 1 to 3 below).

[0085] [Table 1]

[0086] B. Preparation of liposomal antiviral agents by passive loading Liposomes can be prepared by thin-film hydration or solvent injection. The process for preparing liposomal antiviral agents by solvent injection is embodied in a method comprising the following steps. 1. Weigh a predetermined molar ratio of phospholipids and cholesterol lipid mixture in the presence or absence of DSPE-PEG2000, and add them to 10 mL of ethanol in a flask to form a solvent phase containing lipids. 2. A step to prepare a hydroxychloroquine or chloroquine solution in 0.9% sodium chloride (physiological saline) or a suitable medium at a concentration of 40 mg / mL to 60 mg / mL or an appropriate concentration in order to form an aqueous phase. 3. Preheat 40 mL of the indicated aqueous phase (40 mg / mL hydroxychloroquine) at 50°C for at least 30 minutes. 4. The dissolved lipid mixture, i.e., the solvent phase, is added to the preheated aqueous phase by syringe under stirring to form a proliposome sample, and then this proliposome sample is stirred at 50°C for 5 minutes. 5. The above proliposome sample is extruded through a 0.2 μm polycarbonate membrane at an appropriate temperature according to the lipid composition to obtain the designed particle size. 6. The extruded liposome sample is dialyzed and filtered to remove free drug substances with physiological saline (0.9% NaCl), and 7. A step to obtain a liposomal antiviral agent by sterilizing the dialysis-filtered liposome sample by filtering it through a 0.2 μm polycarbonate membrane.

[0087] The antiviral agent compositions relating to this disclosure can be formulated by adding a free antiviral agent to each composition to target an antiviral agent concentration of 1.0 mg / mL to 4 mg / mL and an antiviral agent-to-lipid ratio of at least 0.05 mol / mol to 0.30 mol / mol (see formulation numbers 4 to 6 below).

[0088] [Table 2]

[0089] C. Storage stability of liposomal antiviral agents The stability of liposomal hydroxychloroquine or chloroquine prepared in sections A and B above, stored at 4°C, could be monitored for at least two weeks or for the designed period. Hydroxychloroquine or chloroquine obtained by loading liposomal drug samples into empty liposomes using active or passive loading with ammonium sulfate could be studied. After storing the above liposomal drug samples at 4°C or an appropriate temperature for two weeks or for the designed period, the pharmacokinetics and physicochemical properties of the liposomes could be studied over time.

[0090] Example 2: Preclinical evaluation of inhaled liposomal antiviral agents in animal models. The toxicity of exemplary compositions of liposomal hydroxychloroquine (HCQ) (also known as TLC19) prepared in Example 1B in animals was investigated. A preliminary proof-of-concept pharmacokinetic (PK) and tissue distribution study was performed in Sprague-Dawley (SD) rats after single-dose intravenous (IV) / intratracheal (IT) administration of HCQ sulfate solution (free HCQ) or IT administration of a pilot formulation of TLC19 (test number PK20021). This study was designed to examine the tissue distribution of HCQ, primarily in the lungs, and systemic exposure.

[0091] A total of 52 rats were assigned to three treatment groups. Each rat received a single dose of HCQ via in-vitro or intravenous injection. Organ / tissue samples, including blood and lungs, were collected at pre-specified time points of 0.25 hours, 1 hour (blood only), 4 hours, 24 hours, and 72 hours after administration and used for HCQ determination by liquid chromatography with tandem mass spectrometry. The study design is summarized in Table 1 below.

[0092] [Table 3]

[0093] The concentrations of HCQ in the blood, lungs, and heart were calculated using Analyst® or MassL ynx Software, and the pharmacokinetic (PK) parameters were determined using Phoenix® WinNonlin®. The HCQ concentration-time profiles in the lungs, blood, and heart after administration of the TLC19 pilot formulation and free HCQ are shown in Figures 1, 2, and 3, respectively. The mean PK parameters of HCQ in the lungs, blood, and heart are listed in Table 2.

[0094] [Table 4] a:T max This is shown as the median. b: Units for lungs and heart: μg / g; Units for blood: μg / mL c: Units for lungs and heart: hr * μg / g; unit for blood: hr * μg / mL d: Not applicable

[0095] Regarding lung distribution in rats administered free HCQ, HCQ concentrations steadily and rapidly decreased during the first 24 hours after administration, particularly in the free HCQ IT dose group. Mean HCQ concentrations decreased from 47.8 μg / g to 2.16 μg / g in the IT group and from 9.4 μg / g to 3.77 μg / g in the IV group. In contrast, the degree of HCQ deposition in the lungs was significantly increased in the TLC19 (pilot formulation) group compared to the free HCQ group, which is attributed to the sustained-release properties of the liposomal drug. In the TLC19 group, mean HCQ concentrations decreased from 129 μg / g to 57.1 μg / g during the first 24 hours after administration of half the free HCQ dose, indicating sustained HCQ release in the lungs for a period of time.

[0096] The half-lives of HCQ in the lungs of the free HCQ IV and IT groups were 15.2 hours and 17.7 hours, respectively, which is consistent with the physicochemical properties of HCQ, which allow it to rapidly move freely across cell membranes at physiological pH. The half-life of HCQ in the TLC19 (pilot formulation) group (37.5 hours) was approximately twice that of the free HCQ group. AUC of the TLC19 (pilot formulation) group 0-72 and C max When normalized by dose, lung exposure to TLC19 was 35 times and 29 times higher, respectively, than that of the free HCQ IV group. These results suggest that TLC19 (pilot formulation), a sustained-release formulation of HCQ, successfully extended the lung residence time of HCQ compared to free HCQ administered via IV or IT. Furthermore, our study showed that neither IV nor IT administration of free HCQ could maintain HCQ concentrations in the lungs for an extended period.

[0097] Regarding systemic exposure, HCQ was rapidly absorbed and distributed throughout the system after administration. The T2 levels of HCQ in the blood of the three groups were measured. max The median time was 0.25 hours after administration. max Overall systemic exposure, including AUC, was similar in the free HCQ IV and IT groups. For TLC19 (pilot formulation), the C of HCQ in the blood was similar. max C of free HCQ maxIt was significantly lower than [the other formulation]. A small amount of HCQ resulted in an initial peak concentration within 1 hour after administration of TLC19 (pilot formulation). The remaining HCQ remained in the lungs, allowing for a long residence time and steadily low mean blood concentrations of TLC19 (pilot formulation) between 24 and 72 hours post-administration. The lower blood HCQ concentration levels observed over time suggest that HCQ was released gradually at the local site. As reflected by the longer half-life of TLC19 (pilot formulation), TLC19 (pilot formulation) exhibited a longer release profile than the unformulated HCQ solution.

[0098] HCQ has been shown to cause cardiac disorders, including prolongation of corrected QT time (QTc). To determine the distribution of TLC19 in the heart, the HCQ PK profile in cardiac tissue was determined (Figure 3). TLC19 was found to be more effective at lower cardiac exposure (C) compared to HCQ solution. max ) was observed. When the dose was normalized, similar AUCs were observed in all groups. Considering the lower doses required for local administration (i.e., IT administration) rather than systemic administration (i.e., oral or IV), these results suggest that TLC19 may cause less cardiotoxicity than conventional HCQ administration.

[0099] Two preclinical pharmacokinetic (PK) studies were conducted on TLC19-optimized formulations administered via iodine-transfer (IT) in Sprague Dolly (SD) rats. The details are as follows:

[0100] (1) Single-dose PK study in SD rats: Hemopharmacological and tissue concentration studies of TLC19 after single-dose IT administration were performed in SD rats. Blood and major organs, including lungs, were collected at the planned sampling time points. HCQ concentrations were determined for whole blood and organs. A preliminary PK profile of TLC19 was determined. The percentage of drug distribution of TLC19 in the lungs was also calculated.

[0101] (2) Multiple-dose PK study in SD rats: This study was designed to characterize and evaluate drug accumulation of TLC19 after multiple-dose IT administration. Major organs, including blood and lungs, were collected at planned sampling time points after the first and last doses. The accumulation rate of TLC19 in the blood and lungs could be calculated.

[0102] Example 3: Preparation of liposomal antiviral agents Sulfobutyl ether-β-cyclodextrin (SBECD) was purchased from Zibo Qianhui Biological Technology Co., Ltd., China. SBECD formulation GS-441524 (containing 1.0 mg / mL of GS-441524) was prepared by dissolving GS-441524 (antiviral agent) in a 150 mg / mL SBECD solution with a pH of approximately 4.4, and this was used as test sample (1) GS-441524 solution-IV.

[0103] The test drug (liposomal GS-441524, also known as ISPM21) was prepared by Taiwan Liposome Company, Ltd., Taiwan. It consists of GS-441524 encapsulated in liposomes with an average particle size of approximately 200 nm. GS-441524 was supplied as a pure, pale yellow powder by Formosa Pharmaceuticals, Inc. The liposomes are composed of dipalmitoylphosphatidylcholine (Nippon Seika Co., Ltd., Japan) and cholesterol (Dishman, Netherlands), both of which are natural components of pulmonary surfactant. 23Empty, pre-formed liposomes were prepared by solvent injection. Briefly, an appropriate amount of lipid mixture (1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and cholesterol) was dissolved in ethanol (JT Baker, USA) and injected into an ammonium sulfate solution with stirring at 50°C. The liposome size was adjusted to approximately 200 nm by extruding through a 0.2 μm polycarbonate membrane at 50°C using an extruder. Unencapsulated ammonium sulfate and ethanol were removed by diafiltration to obtain the final empty liposomes.

[0104] Encapsulation of GS-441524 into liposomes was performed using an active loading method. Empty, pre-formed liposomes were mixed with the GS-441524 drug solution and incubated at 50°C to obtain a final ISPM21 sample (liposomal drug suspension) with a pH of 6-7, which was used as test sample (2) ISPM21-IT.

[0105] A. Preparation of liposomal nucleoside compounds I. Preparation of empty liposomes Liposomes could be prepared by thin-film hydration or solvent injection.

[0106] The process for preparing empty liposomes by solvent injection is embodied in a method that includes the following steps: 1. Weighing a lipid mixture of phospholipids and cholesterol in a predetermined molar ratio in the presence or absence of DSPE-PEG2000, and dissolving them in ethanol at a high temperature. 2. A step to prepare a scavenger solution by adding a scavenger (e.g., ammonium sulfate (AS)) to distilled water, mixing the solution, and dissolving the salt. 3. A step of adding the above lipid mixture to the above scavenging agent solution at an appropriate temperature according to the lipid composition to form a liposome solution. 4. The obtained liposome solution is extruded through a polycarbonate membrane at an appropriate temperature according to the lipid composition to obtain the designed particle size. 5. Remove free scavenger and ethanol by dialysfiltration of the extruded liposomes into a sucrose solution, physiological saline, or appropriate buffer solution.

[0107] II. Drug loading of antiviral agents into liposomes The following method represents a typical protocol for encapsulating nucleoside compounds into liposomes by remote loading, and this protocol includes the following steps. 1. A step to obtain a stock solution containing a nucleoside compound (hereinafter referred to as "stock solution") by preparing a solution of a nucleoside compound at a concentration of 15.4 mg / mL or an appropriate concentration in a suitable medium. 2. A process of mixing empty liposomes prepared by the processes of sections (A) and (I) of Example 1 [in a typical embodiment, conditions of a 3:2 molar ratio of DPPC:cholesterol, 300 mM ammonium sulfate (AS) as a scavenging agent, and a phospholipid concentration of 20-50 mM] and the above stock solution together in a conical tube to obtain a loading solution targeting a D / L ratio of 25 g / mol or the designed D / L ratio. 3. The loading solution is continuously shaken at an appropriate temperature for 60 minutes or the designed time to form drug-loaded liposomes. 4. Adding a NaOH solution or buffer solution to the drug-loaded liposomes to adjust the pH of the solution to 6.0-7.0, and 5. A step to determine the drug encapsulation amount (i.e., loading efficiency) of the final sample using size exclusion column chromatography and UV-Vis absorbance measurement or HPLC analysis.

[0108] [Table 5] * Encapsulation efficiency (EE) is calculated by the following formula: EE (%) = LF / TF × 100% = Total drug forms (TF) divided by the liposome forms (LF) of the drug.

[0109] B. Storage stability of liposomal antiviral agents The stability of liposomal nucleoside compounds stored at 4°C could be monitored for at least two months or the designed period. Nucleoside compounds loaded into empty liposomes using 300 mM ammonium sulfate or 75 mM sucrose triethylammonium octasulfate as a scavenger to obtain liposomal drug samples (Table A) could be studied. After storing the liposomal drug samples at 4°C or an appropriate temperature for more than two months or the designed period, the drug efficacy and physicochemical properties of the liposomes could be studied over time.

[0110] Example 4: Release Profile of Liposomal Antiviral Agent In vitro drug release in simulated lung fluid We were able to demonstrate the sustained release characteristics of the liposomal antiviral agents prepared according to Example 1 by conducting release profile experiments. The outline of the in vitro release (IVR) experiment protocol is as follows. 1. Dilute each liposomal antiviral agent sample 10-fold by mixing 0.5 mL with 4.5 mL of SLF (preheated at 37°C), and place the diluted sample in a 15 mL centrifuge tube. 2. Place the centrifuge tube containing the diluted sample on the sample well of the Intelli mixer rotor, rotate at 20 rpm, and incubate at 37°C. 3. To analyze the encapsulation efficiency, a 1 mL diluted sample is taken at a predetermined time point.

[0111] The analytical method for determining the encapsulation efficiency of nucleoside compounds is as follows: a. Pack and wash a 2 mL G50 column with the conditional solution. b. Add 0.1 mL of the sample to the column, then add 0.45 mL of the eluent, and wait for the solution to elute. c. Add 0.8 mL of eluent to the column and collect the eluted material in liposome form. d. Disintegrate the samples (liposome form and whole form) before and after column chromatography using an appropriate solvent. e. Determine the drug concentration of each sample by measuring the absorbance of the sample at the indicated wavelength using UV-Vis or HPLC.

[0112] The encapsulation efficiency (EE) of antiviral agents in liposomes was calculated and obtained using the following formula: Liposome morphology of the drug (LF) divided by total morphology of the drug (TF): EE(%) = LF / TF × 100%

[0113] We were able to plot the release profile and show the release rate (%) versus time. The release rate could be calculated using the following formula: subtract the liposome morphology at each time point from the initial liposome morphology, and then divide by the initial liposome morphology: (LF t0 -LF t ) / LF t0 ×100%

[0114] A long-term release profile of a drug substance is desirable for improved efficacy and treatment at lower dose frequencies. Therefore, the selected liposomal antiviral agents had slower or more appropriate release profiles than all other formulations and were used in the following toxicity studies.

[0115] Example 5: Pharmacokinetics of inhaled liposomal antiviral agents in animal models research design A total of 48 female SD rats (BioLASCO Taiwan Co., Ltd.) were assigned to one of two treatment groups: (1) GS-441524 solution-IV: 24 rats were administered a single dose of 0.20 mg GS-441524 / animal by IV injection; (2) ISPM21-IT: 24 rats were administered a single dose of 0.20 mg ISPM21 (liposomal suspension containing 1.0 mg / mL GS-441524) / animal via IT infusion, an administration route used to mimic inhalation in a clinical setting. Blood samples were sampled at 0.25 hours, 1 hour, 4 hours, 24 hours, and 72 hours post-administration, and lung samples were sampled at 0.25 hours, 4 hours, 24 hours, and 72 hours post-administration. All animal-related procedures were carried out at the TLC animal facility in accordance with the ethical guidelines of TLC and the Institutional Animal Care and Use Committee (IACUC) in Taiwan (number TLC20IACUC037).

[0116] Collection and handling of blood and lung samples Blood was collected from the jugular vein at the scheduled sampling time point into collection tubes containing K2EDTA as an anticoagulant. Each collection tube was gently inverted to ensure complete mixing of the sample with the anticoagulant. The actual sampling time was recorded. The collected blood samples were centrifuged at 1,500 × g for 10 minutes at 2–8°C to obtain plasma. The supernatant plasma was immediately transferred to a labeled microtube. If immediate processing was not required, the plasma was transferred to a freezer set to -80°C. Plasma collection was completed within 2 hours of blood collection.

[0117] The animals were euthanized at the scheduled lung sample collection time. Each rat was perfused with approximately 100 mL of 2 mM K2EDTA / saline solution for at least 8 minutes using a KD Scientific® syringe pump. After perfusion, the lungs were collected and frozen in liquid nitrogen. After freezing, the lungs were weighed and placed on moist ice until transferred to a -80°C freezer. All lung samples were stored in a -80°C freezer until homogenized.

[0118] Biological analysis and PK calculation After adding the internal standard (6,7-dimethyl-2,3-di-2-pyridylquinoxaline) to the blood sample, it was thoroughly mixed with methanol for protein precipitation. After centrifugation, the supernatant was injected into a liquid chromatography system (Waters I-Class UPLC) (LC-MS / MS) connected to a tandem mass spectrometer (Waters Xevo® TQ-S) for analysis. For lung samples, the tissue / organ was homogenized with 50% methanol containing 0.1% formic acid. After adding the internal standard (IS) to the tissue / organ homogenate, it was thoroughly mixed with methanol for protein precipitation. The resulting sample supernatant was injected into an LC-MS / MS for analysis after centrifugation. The concentration of GS-441524 was calculated using MassLynx software. The linear ranges were 10–10000 ng / mL and 0.5–500 ng / mL for the lung assay and plasma assay, respectively. The PK parameters for GS-441524 were calculated using a non-compartment method applying sparse sampling calculations with Phoenix® WinNonlin® (version 8.0 or later).

[0119] result Pharmacokinetics of GS-441524 in the lungs Following a single dose of ISPM21-IT, a longer half-life (22.8 hours) and higher GS-441524 levels in the lungs (Figure 4A) were observed for ISPM21-IT compared to GS-441524 solution-IV (Table 3). The half-life and AUC of GS-441524 solution-IV in the lungs could not be calculated because the concentration was only measurable at the first time point (0.25 hours). In particular, a single dose of 0.2 mg of ISPM21-IT resulted in a C2 level of 74.9 μg / g. max and 369h * AUC of μg / g 0-72 This achieved a 207-fold increase in C compared to GS-441524 solution-IV. max This resulted in dramatically high lung exposure (Table 4).

[0120] [Table 6]

[0121] [Table 7]

[0122] Pharmacokinetics of GS-441524 in plasma After a single dose, ISPM21-IT showed similar PK profiles (Figure 4B) and AUC (Tables 3 and 4) in plasma compared to GS-441524 solution-IV. The plasma half-life of ISPM21-IT (9.98 hours) was slightly longer than that of GS-441524 solution-IV (7.43 hours). In particular, ISPM21-IT showed lower systemic exposure (C) in plasma compared to GS-441524 solution-IV. max This showed 37% (Table 4).

[0123] In this rat PK study, we investigated targeted delivery of inhalable ISPM21 to the lungs and demonstrated sustained release with significantly higher exposure to GS-441524 in the lungs. After a single intra-articular (IT) dose of ISPM21, the mean concentration of GS-441524 in the lungs 72 hours post-administration was 1.07 μg / g (3.67 μM, assuming a lung tissue density of 1 g / mL), which corresponds to 0.18 μM in vitro antiviral EC against SARS-CoV-infected human airway epithelial (HAE) cells in the lungs. 50 This is 19 times higher, indicating that ISPM21 can maintain a relatively high GS-441524 concentration.

Claims

1. A composition of an antiviral agent for use in the prevention or treatment of inhaled respiratory or infectious diseases, comprising a liposomal antiviral agent, wherein the liposomal antiviral agent is A liposome comprising one or more phospholipids and sterols, wherein the molar ratio of total phospholipids to sterols is in the range of 1:1 to 2:1, The antiviral agent encapsulated in the liposomes and Includes, The composition has an antiviral agent to total lipid ratio in the range of 0.01 mol / mol to 2.0 mol / mol, The aforementioned antiviral agent, 【Chemistry 1】 【Chemistry 2】 or selected from the group consisting of these pharmaceutically acceptable salts, Composition of an antiviral agent for use.

2. The antiviral agent composition for use according to claim 1, wherein the sterol is cholesterol.

3. The composition of an antiviral agent for use according to claim 1, wherein the phospholipid is selected from the group consisting of hydrogenated soybean phosphatidylcholine (HSPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), phosphatidylethanolamine lipid, 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), and combinations thereof.

4. The composition of an antiviral agent for use according to claim 1, wherein the liposomal antiviral agent has an average particle size in the range of 50 nm to 1,000 nm.

5. A composition of an antiviral agent for use according to claim 1, further comprising an antibiotic.

6. The antiviral agent composition for use according to claim 5, wherein the antibiotic is selected from the group consisting of curimycin and azithromycin.

7. A composition of an antiviral agent for use according to claim 1, having a total lipid concentration in the range of 1 mM to 100 mM.

8. The antiviral agent composition for use according to claim 1, further comprising a free antiviral agent.

9. The antiviral agent composition for use according to claim 1, having an antiviral agent-to-total lipid ratio in the range of 0.05 mol / mol to 2.0 mol / mol.

10. The composition of an antiviral agent for use according to claim 1, wherein the concentration of the antiviral agent is 0.1 mg / mL to 80 mg / mL.

11. The composition for use of an antiviral agent according to claim 1, wherein the composition is a spray.

12. A particle aerosolized composition comprising an antiviral agent composition according to any one of claims 1 to 11, wherein the particle aerosolized composition is for use in the prevention or treatment of inhaled infectious diseases or respiratory diseases.

13. The particle aerosolization composition according to claim 12, wherein multiple particles have an aerodynamic median particle diameter in the range of 0.5 μm to 5 μm.

14. The composition of an antiviral agent for use according to claim 1, wherein the respiratory disease is selected from the group consisting of severe pneumonia, acute respiratory infections (SARIs) including acute respiratory distress syndrome (ARDS), sepsis, and septic shock.

15. The composition of an antiviral agent for use according to claim 1, wherein the infectious disease is caused by an influenza virus, a retrovirus, a coronavirus, or SARS-CoV-2.

16. A system for administering an antiviral composition to a target that requires it, A composition of an antiviral agent according to any one of claims 1 to 10, A lung delivery device capable of aerosolizing the antiviral agent composition, A system comprising a composition of the aerosolized antiviral agent containing particles containing an amount of free antiviral agent effective to provide immediate antiviral activity and an amount of liposomal antiviral agent effective to provide sustained antiviral activity.

17. The composition for use of an antiviral agent according to claim 1, further comprising a scavenging agent, the scavenging agent comprising an ammonium compound and an anionic counterion, the anionic counterion being selected from the group consisting of sucrose octasulfate anion, dextran sulfate anion, sulfate anion, citrate anion, gluconate anion, sulfonate anion, phosphate anion, pyrophosphate anion, tartrate anion, succinate anion, maleate anion, borate anion, carboxylate anion, bicarbonate anion, glucuronate anion, chloride anion, hydroxide anion, nitrate anion, cyanate anion, bromide anion, and combinations thereof.

18. The composition of an antiviral agent for use according to claim 1, wherein the liposomes further comprise polyethylene glycol (PEG)-modified lipids in an amount ranging from 0.0001 mol% to 10 mol% based on the total lipids.

19. The composition of an antiviral agent for use according to claim 18, wherein the PEG-modified lipid has a PEG portion having an average molecular weight in the range of 1,000 g / mol to 5,000 g / mol.

20. The composition of an antiviral agent for use according to claim 18, wherein the PEG-modified lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)] (DSPE-PEG).

21. The antiviral composition for use according to claim 20, wherein the amount of DSPE-PEG in the liposome is in the range of 0.001 to 5 mol%, based on the total amount of phospholipids and sterols.

22. The antiviral agent composition for use according to claim 10, having a concentration of the antiviral agent in the range of 0.5 to 5 mg / mL.

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