Antiviral compositions for use in the prophylactic or post-exposure treatment of infectious or respiratory diseases

Abstract

Antiviral compositions for use in the prevention or treatment of respiratory or infectious diseases by inhalation are provided. [Solution] A composition comprising a liposomal antiviral agent, the liposomal antiviral agent comprising liposomes containing one or more phospholipids and a sterol, wherein the molar ratio of the total phospholipids to the sterol is in the range of 1:1 to 2:1, and an antiviral agent encapsulated in the liposome, the composition having an antiviral agent to total lipid ratio in the range of 0.01 mol / mol to 2.0 mol / mol, and the antiviral agent being a nucleoside compound of formula (I). TIFF2026035665000016.tif53170

Description

[Technical Field]

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

[0002] Infectious diseases can be transmitted through different infection routes, such as contact infection, droplet infection, and blood-borne infection, and the bioavailability of drugs in an affected physical environment may be low when administered systemically. Penetration of drug delivery systems onto target cells in target tissues is a key obstacle for effectively treating infectious diseases, such as pulmonary infectious diseases, by inhalation. The retention of drug substances within liposomes before the drug delivery system attaches to target epithelial cells may vary from liposomal drug to liposomal drug based on the diffusion rate of free, uncharged drug substances across the lipid membrane of the liposome, which is highly dependent on the physicochemical properties of the lipid barrier in the presence of the external microenvironment of the liposome, as well as the internal aqueous environment of the liposome.

[0003] Respiratory diseases caused by infection or other unknown reasons are extremely severe and debilitating lung diseases that result in premature death, among others, characterized by subsequent efficient viral replication and cell damage caused by virus-induced cell lysis or immunopathology. Infected cell lines and postmortem lung tissues showed cytopathic changes due to apoptosis, necrosis, or sometimes syncytia formation.

[0004] Liposomes have been utilized as drug carriers to mask the unpleasant taste of drugs upon inhalation 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, providing slow drug release in the local physical environment, allowing for less frequent drug administration and optimal dosage, and / or reducing side effects and toxicity. However, it is unknown whether quinine compounds or other antiviral agents delivered by whole liposomes via the inhalation route will effectively perform their required functions, such as deposition on target cell lines expressing suitable receptors as docket sites for viral entry into intracellular target sites, and achieve the desired pharmacokinetic profile in vivo.

[0005] It is not readily apparent that utilizing liposomal technology to reconstitute antivirals would result in liposomal formulations for inhalation at prophylactic doses to prevent severe acute respiratory syndrome or at therapeutic doses to treat respiratory or infectious diseases with reduced side effects. Currently, no practical liposomal drug formulations exist for inhalation as drug products for chemoprophylaxis, such as prevention, treatment of mild cases, or treatment of acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or severe acute respiratory syndrome (SARS), which is caused by infection with viruses such as the coronavirus COVID-19, also known as SARS-CoV2. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] US Patent Application Publication No. 20110104259A1 Summary of the Invention [Problem to be solved by the invention]

[0007] There remains an unmet need for inhalable formulations with a predetermined encapsulation efficiency to achieve a balance between reducing the administration frequency and / or dosage of antiviral agents such as quinine compounds and nucleoside compounds and targeting a desired prophylactic or therapeutic concentration window for pulmonary delivery. In addition, formulations suitable for respiratory diseases should possess properties such as being inhalable, exhibiting sufficient encapsulation efficiency after inhalation administration, having improved stability or suitable resistance to destruction by local substances such as pulmonary surfactants, and having a desired dose strength to ensure the potential for achieving desired efficacy in the pulmonary environment. The present invention addresses this 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 by inhalation, comprising at least one lipid, optionally phospholipid(s) and a sterol, and / or polyethylene glycol (PEG)-modified phospholipid, and an antiviral agent encapsulated in the aqueous interior of the liposome.

[0009] To improve upon existing treatment paradigms for respiratory or infectious diseases and 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 treatment or enhanced treatment of respiratory diseases. In particular, there is a need for an inhalable formulation for the prevention or treatment of SARS.

[0010] The present disclosure provides compositions of antiviral agents for use in the prevention or treatment of respiratory or infectious diseases, particularly SARS, with the following advantages: 1) achieving a longer therapeutic effect compared to inhaled free drug substance; 2) delivering the drug directly to the site of disease or viral infection; 3) more rapid onset of action; 4) reducing adverse drug reactions and systemic effects; 5) bypassing first-pass metabolism observed with oral administration, thus increasing the bioavailability of the drug substance (and potentially reducing cardiotoxicity, ocular symptoms of retinopathy, gastrointestinal (GI) effects including nausea, vomiting, diarrhea, and abdominal discomfort, and hepatotoxicity); 6) increasing the residence time of the drug substance in target tissues via sustained release from liposomal drug; 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 agents of the present disclosure are encapsulated in liposomes in predetermined amounts to form compositions of the antiviral agents of the present disclosure in order to achieve a composition with a favorable release profile and reduced toxicity, particularly cardiac toxicity.

[0012] 1. An antiviral composition for use in treating or preventing a respiratory disease, which is inhalable and comprises a liposomal antiviral agent, the liposomal antiviral agent comprising: a liposome comprising at least one lipid; The antiviral agent encapsulated in this liposome An antiviral composition comprising:

[0013] In some embodiments, the liposome comprises a lipid bilayer composed of one or more phospholipids and a sterol, where the sterol is cholesterol, in a molar ratio of phospholipid(s):cholesterol of 1:1 to 2:1, optionally 3:2.

[0014] In some embodiments, the one or more phospholipids comprise a phosphocholine (PC), which may be, but is not limited to, hydrogenated soybean 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 comprise 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 comprises a 4-aminoquinoline compound.

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

[0017] In some embodiments, the antiviral agent comprises a nucleoside compound of structural formula I: [ka] Each R 1 , R 2 , R 3 , R 4 or R 5 are independently, 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 a are independently H, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, aryl(C1-C8) alkyl, (C4-C8) carbocyclyl alkyl, -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 is H, each X 1 or X 2 independently, CR 10 or N, R 8 is 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) carbocyclyl alkyl, 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 wherein each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Each R 9 or R 10 are independently H, halogen, R 11 , OR 11 , S.R. 11 , N.R. 11 R 12 , N(R 11 )(OR11 ), 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 are independently H, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C4-C8) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)(C1-C8) alkyl, -S(O) n (C1-C8)alkyl or aryl(C1-C8)alkyl, each aryl or heteroaryl independently optionally substituted with one or more Z groups; Or R 11 or R 12 together with the nitrogen to which they are both attached form a 3- to 7-membered heterocycle, and any one carbon atom of the heterocycle may optionally be —O—, —S—, or —NR a - may be replaced by Each Z group independently represents 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 are independently H, alkyl, aryl, arylalkyl, or heterocycle, and one or more non-terminal carbon atoms of each (C1-C8) alkyl are -O-, -S-, or -NR a - may be substituted.

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

[0019] In another aspect, the present disclosure provides an antiviral composition, or an aerosolized composition of particles containing the antiviral composition, for use in the prevention or treatment of an infectious or respiratory disease, the antiviral composition comprising at least 0.01 moles (mol) / mol, and optionally from 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, 0.0 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, and a concentration of antiviral agent in the range of 0.1 mg / mL to 10 mg / mL, are also provided.

[0020] In another aspect, the present disclosure also provides an aerosolized composition of particles comprising a liposomal quinine compound according to the present disclosure for use in preventing or treating a respiratory disease, the aerosolized composition of particles having a drug-to-lipid ratio of at least 0.01 mol / mol, optionally at least 0.05 mol / mol, optionally between 0.01 mol / mol and 2.0 mol / mol, between 0.05 mol / mol and 2.0 mol / mol, between 0.05 mol / mol and 1.5 mol / mol, between 0.05 mol / mol and 1.0 mol / mol, between 0.05 mol / mol and 0.5 mol / mol, between 0.05 mol / mol and 0.3 mol / mol, between 0.05 mol / mol and 0.2 mol / mol, between 0.05 mol / mol and 0.15 mol / mol, optionally about 0.5 mol / mol, and a concentration of quinine compound in the range of 1 mg / mL to 10 mg / mL based on the composition.

[0021] In another aspect, the present disclosure also provides an aerosolized composition of particles comprising a liposomal nucleoside compound according to the present disclosure for use in preventing or treating an infectious disease, the aerosolized composition of particles having a drug-to-lipid ratio of at least 0.01 mol / mol, optionally at least 0.05 mol / mol, optionally between 0.01 mol / mol and 1 mol / mol, 0.03 mol / mol and 0.5 mol / mol, 0.03 mol / mol and 0.15 mol / mol, 0.03 mol / mol and 0.1 mol / mol, optionally about 0.05 mol / mol, 0.05 mol / mol and 0.5 mol / mol, 0.05 mol / mol and 0.15 mol / mol, 0.05 mol / mol and 0.1 mol / mol, 0.07 mol / mol and 0.1 mol / mol, optionally about 0.085 mol / mol, and a concentration of nucleoside compound of between 0.1 mg / mL and 5 mg / mL based on the composition.

[0022] In another aspect, the present disclosure also provides an aerosol spray (spray for inhalation administration) comprising a composition of an antiviral agent for use according to the present disclosure.

[0023] In another aspect, the present disclosure also provides an aerosolized composition of particles containing an antiviral agent composition for use in the prevention or treatment of a respiratory or infectious disease, the aerosolized composition of particles comprising a liposomal antiviral agent according to the present disclosure.

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

[0025] In another aspect, the present disclosure also provides a system for administering an antiviral composition to a subject in need thereof, the system comprising an antiviral composition according to the present disclosure and a pulmonary delivery device capable of aerosolizing the antiviral composition, such that after aerosolization, formed particles containing the liposomal antiviral agent comprise 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 present disclosure also provides a method for reducing complications associated with the treatment of respiratory or infectious diseases in a human subject, the method comprising administering a composition according to the present disclosure to a subject in need thereof. According to the present disclosure, complications include, but are not limited to, cardiotoxicity or hepatotoxicity. According to the present disclosure, complications include, but are not limited to, prolongation of the corrected QT interval (QTc).

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

[0028] [Figure 1] FIG. 1 depicts the pharmacokinetic profiles of HCQ in rat lungs following administration of a composition according to the present disclosure and free HCQ. [Figure 2] FIG. 2 depicts the pharmacokinetic profiles of HCQ in rat blood following administration of a composition according to the present disclosure and free HCQ. [Figure 3] FIG. 3 depicts the pharmacokinetic profiles of HCQ in rat hearts following administration of a composition according to the present disclosure and free HCQ. [Figure 4A]Figures 4A and 4B show a series of graphs depicting the mean concentration-time profiles of GS-441524 in the lungs (Figure 4A) and plasma (Figure 4B) of rats following a single IV dose of SBECD-formulated GS-441524 (GS-441524 Solution-IV) or a single IT dose of liposomal GS-441524, also designated ISPM21 (ISPM21-IT). LLOQ: lower limit of quantitation. [Figure 4B] Figures 4A and 4B show a series of graphs depicting the mean concentration-time profiles of GS-441524 in the lungs (Figure 4A) and plasma (Figure 4B) of rats following a single IV dose of SBECD-formulated GS-441524 (GS-441524 Solution-IV) or a single IT dose of liposomal GS-441524, also designated ISPM21 (ISPM21-IT). LLOQ: lower limit of quantitation. DETAILED DESCRIPTION OF THE INVENTION

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

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

[0031] All numerical values ​​herein may be understood to be modified by "about," which, when referring to measurable values ​​such as amounts, time durations, etc., means to encompass a variation of ±10%, preferably ±5%, more preferably ±1%, and even more preferably ±0.1% from the specified value, since, unless otherwise specified, such a variation is appropriate to obtain the desired amount of liposomal drug.

[0032] As used herein, the terms "treating," "treating," "treated," "treated," "therapy," or "treatment" include preventative (e.g., prophylactic), palliative, and curative uses or results. The term "subject" includes a vertebrate having a respiratory or other disease or suspected of having a viral infection. Preferably, the subject is a warm-blooded animal, including a mammal, preferably a human.

[0033] As used herein, the term "drug" refers to an antiviral agent, such as a quinine compound or a nucleoside compound, associated with activity toward a desired therapeutic effect according to the present 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 a composition according to the present disclosure. The free drug or drug content of a liposomal drug composition according to the present disclosure can be determined by, but is not limited to, ultraviolet-visible absorbance measurement or high-performance liquid chromatography (HPLC) method. The phospholipid content or concentration of liposomes and liposomal drugs can be determined by assaying the phosphorus content of liposome and liposomal drug samples using, but not limited to, a phosphorus assay (adapted from G. Rouser et al., Lipids 1970, 5, 494-496) or an HPLC method.

[0034] As used herein, pharmacokinetic data was obtained in rats, however, pharmacokinetic profiles correlating with inhaled compositions according to the present disclosure can be obtained in other mammals, including but not limited to cats, dogs, horses, mice, pigs, non-human primates, and humans, for the development of inhaled compositions.

[0035] Infectious and respiratory diseases In the present disclosure, infectious diseases and pathogenic infections refer to disorders caused by organisms such as viruses, parasites, and bacteria. In one aspect, infectious diseases are transmitted via fecal-oral, droplet, sexually transmitted, oral, direct contact, fomite, vertical (mother-to-fetus), iatrogenic, or host-borne infection. In another aspect, infectious diseases may include, but are not limited to, urinary tract infections, skin infections, respiratory infections, odontogenic infections, vaginal infections, and intra-amniotic infections.

[0036] In some embodiments, the infectious disease includes acute flaccid osteomyelitis (AFM), anaplasmosis, anthrax, babesiosis, botulism, brucellosis, campylobacteriosis, carbapenem-resistant infections (CRE / CRPA), chancroid, chikungunya virus infection (chikungunya), chlamydia, ciguatera (harmful algal blooms (HABS)), Clostridium difficile infection, Clostridium perfringens (epsilon toxin), coccidioidomycosis (Coccidioidomyces) fungal infection (valley fever), covid-19 (novel coronavirus infection), Creutzfeldt-Jakob disease, Transmissible spongiform encephalopathy (CJD), cryptosporidiosis (Crypto), cyclosporiasis, dengue fever, dengue fever 1, 2, 3, 4 (Dengue), diphtheria, Escherichia coli infection, Shiga toxin-producing (STEC), Eastern equine encephalitis (EEE), Ebola hemorrhagic fever (Ebola), ehrlichiosis, encephalitis, arboviral or post-infectious, enterovirus infection, non-polio (non-polio enterovirus), enterovirus infection D68 (EV-D68), giardiasis (Giardia), glanders, Neisseria gonorrhea (gonorrhea), granuloma inguinale, 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, shingles (Zoster), VZV (shingles), histoplasmosis infection (histoplasmosis), human immunodeficiency virus / AIDS (HIV / AIDS), human papillomavirus (HPV), influenza (flu), lead poisoning, Legionnaires' disease (Legionnaires' disease), leprosy (Leprosy), leptospirosis, listeriosis (Listeria), Lyme disease, lymphogranuloma venereum (LGV), malaria Measles, melioidosis, meningitis, viral (meningitis, viral), meningococcal disease, bacterial (meningitis, bacterial), Middle East Respiratory Syndrome Coronavirus (MERS-CoV), multisystem inflammatory syndrome in children (MIS-C), mumps, norovirus, paralytic shellfish poisoning (paralytic shellfish poisoning, ciguatera), pediculosis (lice, head and body lice), pelvic inflammatory disease (PID), whooping cough (whooping cough), plague (bubonic plague, septicemic plague, pneumonic plague), pneumococcal disease (pneumonia), polio, Powassan, psittacosis (parrot fever), pediculosis (hair lice; groin lice infestation), pustular exanthematous diseases (smallpox, monkeypox, cowpox), Q fever, rabies, ricin poisoning, rickettsiosis (Rocky Mountain spotted fever), rubella (German rubella), including congenitalmeasles), salmonellosis gastroenteritis (Salmonella), scabies mite infestation (Sarcoptes scabiei), scombroid (scombroid food poisoning), septic shock (sepsis), severe acute respiratory syndrome (SARS), shigellosis gastroenteritis (Shigella), smallpox, staphylococcal infection, methicillin-resistant (MRSA), staphylococcal food poisoning, enterotoxin B poisoning (staphylococcal food poisoning), staphylococcal infection, vancomycin-intermediate resistance (VISA), staphylococcal infection, vancomycin-resistant (VRSA), streptococcal disease, group a (invasive) (group A streptococcus (invasive)), streptococcal disease, group b (group B streptococcus), streptococcal toxic shock syndrome, stss, toxic shock (STSS, TSS), syphilis, first stage, second stage These include, but are not limited to, 2-stage, early latent, late latent, congenital, tetanus infection, tetanus (biting spasm), trichomoniasis (trichomoniasis), trichinellosis (trichinosis), tuberculosis (TB), tuberculosis (latent) (LTBI), tularemia (rabbit fever), typhoid fever (group D), typhoid, vaginosis, bacterial (yeast infection), vaping-associated lung injury (e-cigarette-associated lung injury), chickenpox (varicella), cholera (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 according to the present disclosure include, but are not limited to, acute respiratory distress syndrome (ARDS), acute lung injury (ALI), or severe acute respiratory syndrome (SARS), with major complications including fluid leakage into the lungs, making breathing difficult or impossible. Symptoms typically include the gradual onset of fever, cough, productive cough, dry cough, difficulty breathing and fatigue or muscle pain, chest tightness, and shortness of breath. Complications include pulmonary hypertension, heart failure, pneumonia, or pulmonary embolism.

[0038] In some embodiments, the antiviral compositions of the present disclosure are suitable for use in the prophylaxis of, as prevention of, treatment of mild cases of, or treatment 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 antivirals The term "liposome" or "liposomal" as used herein refers to a population of vesicles, each characterized by having an aqueous interior space separated from the external medium by one or more bilayer membranes (bilayer membranes). The liposomal bilayer membrane is typically formed by one or more lipids, i.e., amphiphilic molecules of synthetic or natural origin, containing spatially separated hydrophobic and hydrophilic domains.

[0040] The interior aqueous space of the liposome is substantially free of 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 multilamellar vesicles (MLVs), having average diameters in the ranges of 50 nm to 10,000 nm, 50 nm to 500 nm, 50 nm to 450 nm, 50 nm to 400 nm, 50 nm to 350 nm, 50 nm to 300 nm, 50 nm to 250 nm, 50 nm to 200 nm, 100 nm to 500 nm, 100 nm to 450 nm, 100 nm to 400 nm, 100 nm to 350 nm, 100 nm to 300 nm, 100 nm to 250 nm, or 100 nm to 200 nm, which can pass through a sterilizing filter. For example, MLVs may be formed directly by hydrated lipid films, spray-dried powders, or lyophilized cakes of a selected lipid composition with an entrapment agent, and SUVs and LUVs can be sized from MLVs by sonication, homogenization, microfluidization, or extrusion.

[0041] In general, liposomes typically comprise a lipid mixture comprising at least one lipid selected from the group consisting of dialiphatic chain lipids, such as phospholipids, diglycerides, dialiphatic glycolipids, simple lipids, such as sphingomyelins and glycosphingolipids, sterols, such as cholesterol, and derivatives thereof and combinations thereof.

[0042] Examples of phospholipids according to the present disclosure include 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-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), 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (PSPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), 1,2-distearoyl-sn-glycero-3-phosphatidylcholine (PSPC), 1-distearo ...PSPC), 1-distearoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (PSPC Phosphatidylcholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), hydrogenated soy 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-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) (PSPG), 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'-myoi) Inositol) (ammonium salt) (DPPI), 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 of the present disclosure optionally incorporate polyethylene glycol (PEG)-modified phosphatidylethanolamine (PE) into the membrane of the vesicles, thereby incorporating a significant amount of PEG moieties onto the surface of the vesicles to achieve safe, effective, less frequent administration, and longer, sustained drug release.

[0044] The liposomal antiviral agents of the present disclosure optionally incorporate a significant amount of negatively charged moieties on the surface of the vesicles by incorporating PEG-modified phosphatidylethanolamine (PE) or fatty acids into the vesicle membrane to prevent the liposome aggregation or flocculation (flocculation) process in solution during storage.

[0045] The polyethylene glycol-modified lipid comprises a polyethylene glycol moiety conjugated to 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 lipid is mixed with a phospholipid to form a liposome having one or more bilayer membranes. In some embodiments, the amount of PEG-modified lipid ranges from 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 the total phospholipids and sterols. In some embodiments, the PEG-modified lipid has a PEG moiety with an average molecular weight ranging from 1,000 g / mol to 5,000 g / mol. In certain embodiments, the PEG-modified lipid is a 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 0.01 mol % to 20 mol %, of the total lipid content of the liposome, with the PEG moiety having 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 agent according to the present disclosure includes liposomes having an antiviral agent entrapped therein, which are prepared by encapsulating the antiviral agent in the aqueous interior of the liposomes by a transmembrane pH gradient-driven remote loading method.

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

[0049] In some embodiments, the transmembrane pH gradient is created by using a sequestering agent for remotely loading the antiviral agent into the liposome, the sequestering agent being composed of an ammonium compound and an anionic counterion.

[0050] The term "ammonium compound" refers to NR4 + wherein each R is independently H or an organic residue that is independently alkyl, alkylidene, heterocyclic alkyl, cycloalkyl, aryl, alkenyl, cycloalkenyl, or hydroxyl-substituted derivatives thereof, and optionally contains S, O, or N atoms in the hydrocarbon chain to form ether, ester, thioether, amine, or amide bonds. In one embodiment, the ammonium compound is ammonium.

[0051] The term "anionic counterion" refers to an anionic ion or an entity covalently bound to an anionic functional group. An anionic ion or anionic functional group has a negative charge under physiological conditions.

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

[0053] In one embodiment, the anionic ions and anionic functional groups are selected from one or more of citrate, sulfate, sulfonate, phosphate, pyrophosphate, and carboxylate.

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

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

[0056] In some embodiments, the sequestrant is selected from the group consisting of ammonium sulfate, ammonium phosphate, ammonium citrate, sucrose octasulfate ammonium salt, dextran ammonium sulfate, dimethylammonium sulfate, dimethylammonium phosphate, dimethylammonium citrate, diethylammonium sulfate, diethylammonium phosphate, diethylammonium citrate, sucrose octasulfate diethylammonium salt, dextran diethylammonium sulfate, trimethylammonium sulfate, trimethylammonium phosphate, trimethylammonium citrate, triethylammonium sulfate, triethylammonium phosphate, triethylammonium citrate, sucrose octasulfate triethylammonium salt, dextran triethylammonium 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 includes, 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 a quinine compound, a nucleoside compound, and a combination thereof.

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

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

[0061] Nucleosides have been reported as inhibitors of nonstructural viral proteins, such as RNA-dependent RNA polymerase, and are being used as potential treatments for RNA virus infections. Nucleosides are taken up by cells and converted to triphosphates in vivo, which are expected to compete for the polymerase nucleotide binding site and terminate the polymerase chain reaction. This conversion to triphosphate is usually mediated by cellular kinases, which imposes additional structural requirements on potential nucleoside polymerase inhibitors. The formation of the monophosphate by nucleoside kinases is generally considered the rate-limiting step of the three phosphorylation events. U.S. Patent No. 7,964,580 discloses pronucleosides containing a phosphoramidate moiety masked with a neutral lipophilic group to obtain an appropriate partition coefficient for optimizing cellular uptake and transport, dramatically enhancing the intracellular concentration of nucleoside monophosphate compared to administration of the parent nucleoside alone. Although controversial observations exist, enzyme-mediated hydrolysis of the phosphate moiety likely generates the nucleoside itself instead of the target nucleoside monophosphate immediately after circulation, prior to its targeting to the desired site. For example, via intravenous administration, the known nucleoside compound (2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)oxolane-2-carbonitrile (also known as GS-441524), a metabolite of remdesivir that is more stable 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).Efficient delivery of nucleoside compounds to target cellular sites enriched for key rate-limiting nucleoside kinases may be a universal platform solution to circumvent the complex manufacturing process of different pronucleosides for a wide range of nucleoside compounds.

[0062] In some embodiments, the antiviral compositions of the present disclosure further comprise an antibiotic, a supplement, or a combination thereof.

[0063] In some embodiments, the antiviral agent comprises one or more 1'-substituted carba-nucleoside compounds or 2'-substituted carba-nucleoside compounds described in U.S. Pat. Nos. 8,008,264 and 9,481,704.

[0064] In some embodiments, the antiviral agents are directed to nucleoside compounds, including but not limited to, 1'-substituted carba-nucleoside compounds and pharmaceutically acceptable salts thereof.

[0065] In some embodiments, the antiviral agent comprises an inhibitor of RNA-dependent RNA viral polymerase, which comprises a nucleoside compound of structural formula (I): [ka] Each R 1 , R 2 , R 3 , R 4 or R 5 are independently, 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 aare independently H, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, aryl(C1-C8) alkyl, (C4-C8) carbocyclyl alkyl, -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 is H, each X 1 or X 2 independently, CR 10 or N, R 8 is 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) carbocyclyl alkyl, 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 wherein each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Each R 9 or R 10 are independently H, halogen, R 11 , OR 11 , S.R. 11 , N.R. 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 are independently H, (C1-C8) alkyl, (C2-C8) alkenyl, (C2-C8) alkynyl, (C4-C8) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, -C(=O)(C1-C8) alkyl, -S(O) n (C1-C8)alkyl or aryl(C1-C8)alkyl, each aryl or heteroaryl independently optionally substituted with one or more Z groups; Or R 11 or R 12 together with the nitrogen to which they are both attached form a 3- to 7-membered heterocycle, and any one carbon atom of the heterocycle may optionally be —O—, —S—, or —NR a - may be replaced by Each Z group independently represents 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 are independently H, alkyl, aryl, arylalkyl, or heterocyclic ring, and one or more non-terminal carbon atoms of each of the (C1-C8) alkyls are -O-, -S-, or -NR a - may be substituted.

[0066] In some embodiments, the antiviral agent is [ka] [ka] or a pharmaceutically acceptable salt thereof.

[0067] In one embodiment, the liposomal antiviral agent comprises a lipid bilayer comprising one or more phospholipids, a sterol, and optionally a polyethylene glycol (PEG)-modified lipid, particularly a PEG-modified phosphatidylethanolamine (PEG-PE), and an aqueous interior encompassed by the 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 being in the range of 0.001 to 5 mol%, optionally 0.0001 mol% to 40 mol%, optionally less than 6 mol%, and optionally 0.001 mol% to 30 mol%, based on the total phospholipids and sterols.

[0069] In one embodiment, the liposomal antiviral 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 includes a lipid bilayer comprising DPPC and cholesterol, and an aqueous interior encompassed by the lipid bilayer and containing an antiviral agent entrapped by a sequestering agent, wherein the antiviral agent is (2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)oxolane-2-carbonitrile (GS-441524), and the sequestering agent is ammonium sulfate.

[0070] Inhalable (inhalation) compositions and aerosolized particles thereof The antiviral agent composition of the present disclosure is adapted to prepare an inhalable aerosolized composition of particles containing the liposomal antiviral agent described above. This composition can be administered for inhalation as a nebulized spray or aerosol, or by intrathecal administration. Inhalation administration is preferred. The overall result is less frequent administration and a higher therapeutic index compared to the free drug or parenteral forms of the drug. The liposomal antiviral agents in this composition are particularly advantageous due to their ability to protect the drug while being compatible with the lung lining or lung surfactant.

[0071] In one embodiment, the antiviral composition of the present disclosure has a drug-to-lipid ratio (D / L) 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 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 composition has at least one lipid at a concentration 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 composition has one or more phospholipids at a concentration 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 quinine compounds 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 composition has one or more phospholipids at a concentration 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, and a drug-to-lipid (D / L) ratio ranging from 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 carba-nucleoside compound or a 2'-substituted carba-nucleoside compound having a free 5'-OH group.

[0075] In some embodiments, the liposomal antiviral composition further comprises a free antiviral agent, wherein the free antiviral agent in the compositions of the present disclosure is 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 in the range of 10% to 40%, 15% to 35%, or 10% to 30% of the total amount of antiviral agent in the composition.

[0076] In some embodiments, the aerosolized composition of particles containing the composition of the present disclosure is generated by aerosolizing the composition using a nebulizer selected from the group consisting of an air-jet nebulizer, an ultrasonic nebulizer, a vibrating mesh nebulizer, a condensation aerosol generator, an electrohydrodynamic nebulizer, or other pulmonary delivery device known in the art.

[0077] In some embodiments, the particles of the aerosolized composition have a mass median aerodynamic diameter of from 0.5 μm to 5 μm, optionally from 1 μm to 3 μm.

[0078] After aerosolization, leakage of the antiviral agent from the liposomes of the liposomal antiviral agent in the composition of the present disclosure results in a portion of the antiviral agent being free and not entrapped 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%, optionally in a controlled percentage ranging from 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%, or 25% to 35%.

[0079] In a specific embodiment, an aerosolized composition of the particles is subjected to pulmonary delivery to a subject requiring a release rate of about 0.5%-25% of the administered drug dose per hour, with complete release of the antiviral agent occurring after a minimum of about 12-24 hours.

[0080] The present disclosure is further described with reference to the following specific, non-limiting examples. [Example]

[0081] The following examples illustrate the preparation and properties of certain embodiments of the present disclosure.

[0082] Example 1 Stability of liposomal antiviral agents Liposomal antiviral agents according to the present disclosure include liposomes with entrapped antiviral agents prepared by active loading or passive loading methods known in the art.

[0083] A. Preparation of Liposomal Antiviral Agents by Active Loading I. Preparation of empty liposomes The process of preparing empty liposomes for remote loading was carried out by thin film hydration method or solvent injection. These methods may include 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 adding them to 10 mL of chloroform in a round-bottom flask; 2. Attaching the flask to a rotary evaporator at an appropriate temperature depending on the lipid composition, stirring the flask to dissolve the lipid mixture, and then placing the flask under vacuum with stirring to evaporate the chloroform and obtain a dry lipid film; 3. Preparing a scavenger solution (e.g., ammonium sulfate (AS)) by adding a scavenger to distilled water and vortexing the solution to dissolve the powder; 4. Adding the above-mentioned capture agent solution to the dried lipid thin film and stirring at an appropriate temperature depending on the lipid composition to form a liposome solution; 5. Freezing and thawing the liposome solution using liquid nitrogen and a water bath at an appropriate temperature depending on the lipid composition to obtain a liposome sample; 6. Extrusion of the obtained liposome sample through a 0.2 μm polycarbonate membrane and a 0.1 μm polycarbonate membrane at an appropriate temperature depending on the lipid composition to obtain the designed particle size; 7. Dialyzing the extruded liposome sample to remove free scavenger, then adding the sample to a dialysis bag (MWCO: 25 kD), sealing the bag, and stirring the dialysis bag in 100x volume of 9.4% (w / v) sucrose solution or saline or a suitable buffer, and then replacing the sucrose solution or saline or a suitable buffer after 1 hour and 4 hours, and stirring overnight; 8. Sterilizing the dialyzed liposome sample by filtering through a 0.45 μm PTFE membrane to obtain empty liposomes.

[0084] II. Drug loading of antiviral agents into liposomes to obtain liposomal antiviral agents The following method represents a typical protocol for the encapsulation of hydroxychloroquine or chloroquine into liposomes by remote loading, which includes the following steps: 1. preparing a solution of 40 mg / mL or an appropriate concentration of hydroxychloroquine or chloroquine in 9.4% (w / v) sucrose or an appropriate vehicle, 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. Mixing together empty liposomes prepared by the process according to Section A(I) of Example 1 (in a typical embodiment, using conditions of a 3:2 DPPC:cholesterol molar ratio, 300 mM ammonium sulfate (AS), and 30 mM phospholipid concentration), saline, and stock solution 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. Continuously shaking the loading solution at an appropriate temperature for 30 minutes or a designed time period to form a drug-loaded liposome sample; 4. If necessary, remove free drug or change the buffer or adjust the drug concentration by dialysis or membrane-based tangential flow filtration (TFF); and 5. Determine the drug loading (i.e., loading efficiency) of the final sample using size exclusion column chromatography and HPLC analysis to obtain liposomal antiviral compositions with drug concentrations ranging from 2 mg / mL to 10 mg / mL and antiviral to lipid ratios of 0.05 mol / mol to 1.5 mol / mol, based on the total composition (see Formulations Nos. 1 to 3 below).

[0085] [Table 1]

[0086] B. Preparation of Liposomal Antivirals by Passive Loading Liposomes can be prepared by thin film hydration or solvent infusion. The process for preparing liposomal antiviral agents by solvent infusion is embodied by a method comprising 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 adding it to 10 mL of ethanol in a flask to form a lipid-containing solvent phase; 2. Preparing a hydroxychloroquine or chloroquine solution in 0.9% sodium chloride (saline) or a suitable vehicle at 40 mg / mL to 60 mg / mL or an appropriate concentration to form an aqueous phase; 3. Preheating 40 mL of the indicated aqueous phase (40 mg / mL hydroxychloroquine) to 50° C. for at least 30 minutes; 4. Adding the dissolved lipid mixture, i.e., the solvent phase, to the preheated aqueous phase via syringe under stirring to form a proliposomal sample, and then continuing to stir the proliposomal sample at 50°C for 5 minutes; 5. Extruding the proliposome sample through a 0.2 μm polycarbonate membrane at an appropriate temperature depending on the lipid composition to obtain the designed particle size; 6. Diafiltration of the extruded liposome sample to remove free drug substance with saline (0.9% NaCl); and 7. Sterilizing the diafiltered liposome sample by filtering through a 0.2 μm polycarbonate membrane to obtain the liposomal antiviral agent.

[0087] Antiviral compositions according to the present disclosure can be formulated by adding free antiviral agent to target concentrations of antiviral agent between 1.0 mg / mL and 4 mg / mL, and ratios of antiviral agent to lipid of at least 0.05 mol / mol to 0.30 mol / mol, each based on the total composition (see Formulations Nos. 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 a designated period. Hydroxychloroquine or chloroquine loaded into blank liposomes by active loading with ammonium sulfate or by passive loading to obtain liposomal drug samples could be studied. After storing the liposomal drug samples at 4°C or at an appropriate temperature for two weeks or for a designated period, the drug potency and physicochemical properties of the liposomes could be studied over time.

[0090] Example 2 Preclinical Evaluation of Inhaled Liposomal Antivirals in Animal Models The toxicity of the exemplary liposomal hydroxychloroquine (HCQ) composition (also designated TLC19) prepared in Example 1B in animals was investigated. One preliminary proof-of-concept pharmacokinetic (PK) and tissue distribution study was conducted in Sprague-Dawley (SD) rats after a single-dose intravenous (IV) / intratracheal (IT) administration of HCQ sulfate solution (free HCQ) or IT administration of a pilot formulation of TLC19 (Study No. PK20021). This study was designed to examine tissue distribution, primarily pulmonary, and systemic exposure of HCQ.

[0091] A total of 52 rats were assigned to three treatment groups. Each rat received a single dose of HCQ via IT administration or IV injection. Organ / tissue samples, including blood and lung, were collected at pre-specified time points: 0.25 hours, 1 hour (blood only), 4 hours, 24 hours, and 72 hours post-dose 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] Concentrations of HCQ in the blood, lungs, and heart were calculated by using Analyst® or MassLynx Software, and pharmacokinetic (PK) parameters were determined using Phoenix® WinNonlin®. HCQ concentration versus 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 is shown as the median. b: Units for lung and heart: μg / g; Units for blood: μg / mL c: Unit 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 declined during the first 24 hours after administration, particularly in the free HCQ IT group. The mean HCQ concentration 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 extent of HCQ deposition in the lungs was significantly increased in the TLC19 (pilot formulation) group compared with the free HCQ group, which is attributed to the sustained-release properties of the liposomal drug. The TLC19 group sustained HCQ release in the lungs for a period of time, as the mean HCQ concentration decreased from 129 μg / g to 57.1 μg / g during the 24 hours after the first administration after administration of half the free HCQ dose.

[0096] The half-lives of HCQ in the lungs in the free HCQ IV and IT groups were 15.2 and 17.7 hours, respectively, which is consistent with the physicochemical properties of HCQ, which allows HCQ to freely move across cell membranes rapidly 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. The AUC of the TLC19 (pilot formulation) group 0-72 and C max The lung exposures for the IV and IT groups were 35- and 29-fold higher than those for the IV free HCQ group, respectively, when normalized by dose. These results suggest that the sustained-release formulation of HCQ, TLC19 (pilot formulation), successfully extended the lung residence time of HCQ compared with free HCQ administered IV or IT. Furthermore, in our study, neither IV nor IT administration of free HCQ was able to maintain HCQ concentrations in the lung for an extended period of time.

[0097] Regarding systemic exposure, HCQ was rapidly absorbed and distributed throughout the system after administration. max The median time was 0.25 hours after administration. max Overall systemic exposure, including C and AUC, was similar in the free HCQ IV and IT groups. For TLC19 (pilot formulation), the C of HCQ in blood was max is the C of free HCQ maxThe HCQ concentration was significantly lower than that of the unformulated HCQ solution. The 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 a steadily low mean blood concentration of TLC19 (pilot formulation) between 24 and 72 hours after administration. The lower blood HCQ concentration levels observed over time suggest that HCQ was gradually released 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 damage, including prolongation of the corrected QT interval (QTc). To determine the distribution of TLC19 in the heart, the HCQ PK profile in cardiac tissue was determined (Figure 3). TLC19 resulted in lower cardiac exposure (C) when compared to HCQ solution. max ) when normalized for dose, 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 PK studies were conducted on the optimized TLC19 formulation by IT administration in Sprague-Dawley (SD) rats, as described below.

[0100] (1) Single-dose PK study in SD rats: Blood pharmacokinetics and tissue concentration studies after a single IT dose of TLC19 were conducted in SD rats. Blood and major organs, including lungs, were collected at scheduled sampling times. HCQ concentrations were determined in 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 the drug accumulation of TLC19 after multiple IT doses. At scheduled sampling time points after the first and last doses, blood and major organs, including lungs, could be collected. The accumulation rates of TLC19 in blood and lungs could be calculated.

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

[0103] The test drug (liposomal GS-441524, also referred to 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 provided by Formosa Pharmaceuticals, Inc. as a pure, pale yellow powder. The liposomes were composed of dipalmitoylphosphatidylcholine (Nippon Fine Chemical Co., Ltd., Japan) and cholesterol (Dishman, The Netherlands), both of which are natural components of pulmonary surfactant. 23Empty preformed 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 extrusion at 50 °C through a 0.2 μm polycarbonate membrane 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 the active loading method: empty preformed liposomes were mixed with GS-441524 drug solution and incubated at 50 °C to obtain the final ISPM21 sample (liposomal drug suspension) with a pH of 6–7, which was used as test article (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 infusion methods.

[0106] The process for preparing empty liposomes by solvent infusion is embodied by a method comprising 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 it in ethanol at high temperature; 2. Preparing a scavenger solution by adding a scavenger (e.g., ammonium sulfate (AS)) to distilled water and mixing the solution to dissolve the salt; 3. Adding the lipid mixture to the capture agent solution at an appropriate temperature depending on the lipid composition to form a liposome solution; 4. The resulting liposome solution is extruded through a polycarbonate membrane at an appropriate temperature depending on the lipid composition to obtain the designed particle size. 5. Diafiltration of the extruded liposomes against a sucrose solution or saline or suitable buffer to remove free scavenger and ethanol.

[0107] II. Drug loading of antiviral drugs into liposomes The following method represents a typical protocol for encapsulation of nucleoside compounds into liposomes by remote loading, which comprises the following steps: 1. Preparing a solution of 15.4 mg / mL or an appropriate concentration of a nucleoside compound in a suitable medium to obtain a stock solution containing the nucleoside compound (hereinafter referred to as stock solution); 2. Mixing empty liposomes prepared by the process according to Section (A) and Section (I) of Example 1 (in a typical embodiment, conditions of a 3:2 DPPC:cholesterol molar ratio, 300 mM ammonium sulfate (AS) as a sequestering agent, and a phospholipid concentration of 20-50 mM) and the above stock solution together in a conical tube to obtain a loading solution with a D / L ratio of 25 g / mol or a designed D / L ratio; 3. Continuously shaking the loading solution at an appropriate temperature for 60 minutes or a designed time period to form drug-loaded liposomes; 4. Adding a NaOH solution or a buffer solution to the drug-loaded liposomes to adjust the pH of the solution to 6.0-7.0; and 5. Determining the drug loading (i.e., loading efficiency) of the final sample using size exclusion column chromatography and UV-visible absorbance measurement or HPLC analysis.

[0108] [Table 5] * The encapsulation efficiency (EE) is calculated by the following formula: liposomal form of drug (LF) divided by total form of drug (TF): EE (%) = LF / TF × 100%.

[0109] B. Storage stability of liposomal antiviral agents The stability of liposomal nucleoside compounds stored at 4°C could be monitored for at least 2 months or for a designed period. Nucleoside compounds loaded into blank liposomes using 300 mM ammonium sulfate or 75 mM sucrose octasulfate triethylammonium salt as a scavenger to obtain liposomal drug samples (Table A) could be studied. After storing the liposomal drug samples at 4°C or at the appropriate temperature for at least 2 months or for a designed period, the drug potency and physicochemical properties of the liposomes could be studied over time.

[0110] Example 4 Release Profile of Liposomal Antiviral Agents In vitro drug release in simulated lung fluid Release profile experiments were carried out on the liposomal antiviral agents prepared according to Example 1 to demonstrate their sustained release properties. The protocol for the in vitro release (IVR) experiment is outlined below. 1. Dilute the test product 10-fold by mixing 0.5 mL of each liposomal antiviral sample with 4.5 mL of SLF (preheated to 37°C), and place the diluted sample in a 15 mL centrifuge tube. 2. Place the centrifuge tube containing the diluted sample onto the sample well of the Intelli Mixer rotor, rotate at 20 rpm, and incubate at 37°C. 3. Sample 1 mL of diluted sample at predetermined time points to analyze encapsulation efficiency.

[0111] The analytical method for determining the encapsulation efficiency of the nucleoside compounds is as follows. a. Pack and wash a 2 mL G50 column with the conditioned solution; b. Add 0.1 mL of sample to the column, then add 0.45 mL of eluent and wait for the solution to elute; c. Add 0.8 mL of eluent to the column and collect the eluate in liposome form; d. Disrupt the pre-column and post-column samples (liposomal and total forms) with an appropriate solvent; e. Determine the drug concentration of each sample by measuring the absorbance of the samples at the indicated wavelengths using UV-Vis or HPLC methods.

[0112] The encapsulation efficiency (EE) of the antiviral agent in liposomes was calculated and could be obtained by the following formula: liposomal form of drug (LF) divided by total form of drug (TF): EE(%)=LF / TF×100%

[0113] The release profile could be plotted to depict the release rate (%) versus time. The release rate could be calculated by the following formula: initial liposome morphology minus liposome morphology at each time point, then divided by the initial liposome morphology: (LF t0 -LF t ) / LF t0 ×100%

[0114] A prolonged release profile of a drug substance is desirable for improved efficacy and less frequent administration of treatment. Therefore, the liposomal antiviral agents selected had the slowest or most appropriate release profile of all formulations and were used in the following toxicity studies.

[0115] Example 5 Pharmacokinetics of inhaled liposomal antivirals 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 received a single dose of 0.20 mg GS-441524 / animal via IV injection; (2) ISPM21-IT: 24 rats received a single dose of 0.20 mg ISPM21 (a liposomal suspension containing 1.0 mg / mL GS-441524) / animal via IT instillation, a route of administration used to mimic inhalation in clinical settings. Blood samples were collected at 0.25, 1, 4, 24, and 72 hours post-dose, and lung samples were collected at 0.25, 4, 24, and 72 hours post-dose. All procedures involving animals were performed in the TLC animal facility in accordance with the ethical guidelines of the Institutional Animal Care and Use Committee (IACUC) at TLC, Taiwan (number TLC20IACUC037).

[0116] Collection and handling of blood and lung samples Blood was collected from the jugular vein at scheduled sampling times into collection tubes containing K2EDTA as an anticoagulant. Each collection tube was gently inverted to ensure the sample was thoroughly mixed 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 labeled microtubes. If not processed immediately, the plasma was transferred to a freezer set at -80°C. Plasma collection was completed within 2 hours of blood collection.

[0117] At the scheduled lung sampling time, animals were euthanized. 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, lungs were harvested and frozen in liquid nitrogen. After freezing, lungs were weighed and placed on wet ice until transferred to a -80°C freezer. All lung samples were stored in a -80°C freezer until homogenization.

[0118] Bioanalysis and PK calculations An internal standard (6,7-dimethyl-2,3-di-2-pyridylquinoxaline) was added to the blood samples, which were then thoroughly mixed with methanol for protein precipitation. After centrifugation, the supernatant was injected into a liquid chromatography (Waters I-Class UPLC) coupled to a tandem mass spectrometer (Waters Xevo™ TQ-S) (LC-MS / MS) for analysis. For lung samples, tissues / organs were homogenized in 50% methanol containing 0.1% formic acid. An internal standard (IS) was added to the tissue / organ homogenate, which was then thoroughly mixed with methanol for protein precipitation. The resulting sample supernatant was injected into the LC-MS / MS for analysis after centrifugation. The concentration of GS-441524 was calculated using MassLynx software. The linear ranges were 10-10,000 ng / mL and 0.5-500 ng / mL for the lung and plasma assays, respectively. PK parameters of GS-441524 were calculated by non-compartmental methods applying sparse sampling calculations using Phoenix® WinNonlin® (version 8.0 or higher).

[0119] result GS-441524 Pharmacokinetics in the Lungs After a single IT dose of ISPM21, a longer half-life (22.8 hours) and higher GS-441524 levels in the lung (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 lung could not be calculated because concentrations were measurable only at the first time point (0.25 hours). Notably, a single dose of 0.2 mg of ISPM21-IT resulted in a C of 74.9 μg / g. max and 369h * AUC in μg / g 0-72 Achieved 207-fold higher C than GS-441524 solution-IV max showed dramatically higher lung exposures (Table 4).

[0120] [Table 6]

[0121] [Table 7]

[0122] GS-441524 pharmacokinetics in plasma After single-dose administration, ISPM21-IT showed similar PK profiles (Figure 4B) and AUCs (Tables 3 and 4) in plasma compared with 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). Notably, ISPM21-IT showed lower systemic exposure (C) in plasma compared with GS-441524 solution-IV. max 37% of the total (Table 4).

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

Claims

1. 1. An antiviral composition for use in the prevention or treatment of a respiratory disease or an infectious disease, comprising a liposomal antiviral agent, the liposomal antiviral agent comprising: a liposome comprising at least one lipid; an antiviral agent encapsulated in the liposome; 10. An antiviral composition for use comprising:

2. 2. The antiviral composition for use according to claim 1, wherein the infectious disease is caused by a viral infection and optionally a coronavirus infection.

3. 2. The antiviral composition for use according to claim 1, wherein the at least one lipid comprises one or more phospholipids and a sterol, and the molar ratio of total phospholipids to sterol is in the range of 1:1 to 2:

1.

4. 4. The antiviral composition for use according to claim 3, wherein said sterol is cholesterol.

5. 4. The antiviral composition for use according to claim 3, wherein the at least one lipid is selected from the group consisting of hydrogenated soy phosphatidylcholine (HSPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), phosphatidylethanolamine lipids such as 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE), and combinations thereof.

6. 2. The antiviral composition for use according to claim 1, wherein the liposomal antiviral agent has an average particle size in a range selected from the group consisting of 50 nm to 1,000 nm, 50 nm to 500 nm, 100 nm to 300 nm, and 150 nm to 250 nm.

7. 2. The antiviral composition for use according to claim 1, wherein said antiviral agent is selected from the group consisting of quinine compounds, nucleoside compounds and combinations thereof.

8. 2. The antiviral composition for use according to claim 1, wherein the antiviral agent is selected from the group consisting of hydroxychloroquine, chloroquine, amodiaquine and pharmaceutically acceptable salts thereof.

9. the antiviral agent is a nucleoside compound of structural formula I, 【Chemistry 1】 Each R １ , R ２ , R ３ , R ４ or R ５ are independently H, OR ａ , N(R ａ ) ２ , N ３ , C.N., N.O. ２ , S(O) ｎ R ａ , halogen or methyl, and n is 0, 1 or 2; R ６ is CN or H, Each R ａ are independently H, (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, aryl (C １ ~C ８ ) alkyl, (C ４ ~C ８ ) carbocyclylalkyl, —C(═O)R １１ , -C(=O)OR １１ , —C(═O)NR １１ R １２ , -C(=O)SR １１ , -S(O)R １１ , -S(O) ２ R １１ , -S(O)(OR １１ ), -S(O) ２ (OR １１ ), or -SO ２ NR １１ R １２ and R ７ is H, each X １ or X ２ are independently, C-R １０ or N, R ８ is halogen, NR １１ R １２ , N(R １１ ) (OR １１ ), N.R. １１ NR １１ R １２ , N ３ , NO, NO ２ , 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 , (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, aryl (C １ ~C ８ ) alkyl, (C ４ ~C ８ ) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)(C １ ~C ８ ) alkyl, —S(O) ｎ (C １ ~C ８ ) alkyl, aryl (C １ ~C ８ ) alkyl, OR １１ or SR １１ wherein each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Each R ９ or R １０ are independently H, halogen, R １１ , OR １１ , S.R. １１ , N.R. １１ R １２ , N(R １１ ) (OR １１ ), N.R. １１ NR １１ R １２ , N ３ , NO, NO ２ , 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 are independently H, (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, (C ４ ~C ８ ) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)(C １ ~C ８ ) alkyl, —S(O) ｎ (C １ ~C ８ ) alkyl or aryl (C １ ~C ８ ) alkyl, and each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Or, R １１ or R １２ together with the nitrogen to which they are both attached form a 3- to 7-membered heterocycle, and any one carbon atom of said heterocycle is optionally —O—, —S—, or —NR ａ - may be replaced by Each Z group independently represents 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, -NO ２ , = N ２ , -N ３ , -NHC(=O)R ｂ , -OC(=O)R ｂ , -NHC(=O)NR ｂ ２ , -S(=O) ２ -, -S(=O) ２ OH, -S(=O) ２ R ｂ , -OS(=O) ２ OR ｂ , -S(=O) ２ OH, -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 are independently H, alkyl, aryl, arylalkyl, or heterocycle, and each of the above (C １ ~C ８ ) One or more of the non-terminal carbon atoms of the alkyl is —O—, —S—, or —NR ａ may be substituted with - 10. An antiviral composition for use according to claim 1.

10. 10. An antiviral composition for use according to claim 1, claim 8 or claim 9, further comprising an antibiotic.

11. 11. The antiviral composition for use according to claim 10, wherein said antibiotic is selected from the group consisting of curimycin and azithromycin.

12. 10. The antiviral composition for use according to claim 1, claim 8 or claim 9, having a lipid concentration in a range selected from the group consisting 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 and 20 mM to 60 mM.

13. 13. An antiviral composition for use according to any one of claims 1 to 12, further comprising a free antiviral agent.

14. 13. The antiviral composition for use according to any one of claims 1 to 12, having a drug to lipid ratio in the range selected from the group consisting of: 0.01 mol / mol to 2.0 mol / mol, 0.05 mol / mol to 2.0 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, 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, 0.01 mol / mol to 1 mol / mol, 0.05 mol / mol to 0.15 mol / mol, and 0.05 mol / mol to 0.1 mol / mol.

15. 13. The composition of any one of claims 1 to 12, wherein the antiviral agent is hydroxychloroquine or a pharmaceutically acceptable salt thereof, and the total amount of the antiviral agent is from about 0.1 mg / mL to about 80 mg / mL, from about 0.1 mg / mL to about 80 mg / mL, from about 0.5 mg / mL to about 60 mg / mL, from about 0.5 to about 30 mg / mL, and from about 0.5 mg / mL to about 15 mg / mL, from about 0.5 mg / mL to about 10 mg / mL, from about 0.5 mg / mL to about 8 mg / mL, from about 1.0 mg / mL to about 6 mg / mL, from about 1.5 mg / mL to about 5.0 mg / mL, from about 1.5 mg / mL to about 3.0 mg / mL, or about 2 mg / mL.

16. 1. An antiviral composition for use in the prevention or treatment of a respiratory disease or an infectious disease, comprising a liposomal antiviral agent and a free antiviral agent, wherein the liposomal antiviral agent is a liposome comprising one or more phospholipids and cholesterol; the liposome-encapsulated hydroxychloroquine, chloroquine, or (2R,3R,4S,5R)-2-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(hydroxymethyl)oxolane-2-carbonitrile; Including, 1. A composition of an antiviral agent for use, wherein the composition has a total concentration of the antiviral agent in the range of 0.5 mg / mL to 15 mg / mL, 0.5 mg / mL to 5 mg / mL, or 0.5 mg / mL to 4 mg / mL, a lipid concentration in the range of 5 mM to 60 mM, and a drug to lipid ratio in the range of 0.01 mol / mol to 2 mol / mol, 0.01 mol / mol to 0.3 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, or 0.05 mol / mol to 0.15 mol / mol.

17. 10. The antiviral composition for use according to claim 1, wherein the composition is an aerosol spray.

18. 17. An aerosolized composition of particles comprising the antiviral composition of any one of claims 1 to 16 for use in the prevention or treatment of an infectious or respiratory disease by inhalation, said antiviral composition comprising: a liposome comprising at least one lipid; an antiviral agent encapsulated in the liposome; 1. An aerosolized composition of particles comprising:

19. 20. The aerosolized composition of claim 18, wherein the plurality of particles have a mass median aerodynamic diameter in the range of about 0.5 μm to 5 μm.

20. 1. A method for treating or preventing an infectious or respiratory disease, comprising: Administering to a subject in need thereof an effective amount of an antiviral composition for use according to any one of claims 1 to 16. A method comprising:

21. 21. The method of claim 20, wherein the respiratory disease is selected from the group consisting of severe pneumonia, acute respiratory infection (SARI) including acute respiratory distress syndrome (ARDS), sepsis, and septic shock.

22. 21. The method of claim 20, wherein the infectious disease is caused by a virus, optionally an influenza virus or a retrovirus, optionally a coronavirus, and optionally SARS-Co V-2.

23. 1. A system for administering an antiviral composition to a subject in need thereof, comprising: An antiviral composition according to any one of claims 1 to 16; a pulmonary delivery device capable of aerosolizing the antiviral composition; wherein the aerosolized composition of the antiviral agent comprises particles containing 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.

24. 1. An antiviral pharmaceutical composition for use in treating an infectious disease, comprising a liposomal antiviral agent, the liposomal antiviral agent comprising: a liposome comprising at least one lipid; an antiviral agent encapsulated in the liposome by remote loading using a scavenger; wherein the sequestering agent is comprised of an ammonium compound and an anionic counterion, and the anionic counterion is selected from the group consisting of sucrose octasulfate, dextran sulfate, sulfate, citrate, gluconate, sulfonate, phosphate, pyrophosphate, tartrate, succinate, maleate, borate, carboxylate, bicarbonate, glucuronate, chloride, hydroxide, nitrate, cyanate, bromide, and combinations thereof.

25. 25. The pharmaceutical composition of an antiviral agent for use according to claim 24, wherein the lipid bilayer comprises polyethylene glycol (PEG)-modified lipids in an amount ranging from 0.0001 mol% to 10 mol%, optionally less than 6 mol%, optionally ranging from 0.001 mol% to 5 mol%, based on the total lipids.

26. 26. The pharmaceutical composition of an antiviral agent for use according to claim 25, wherein the PEG-modified lipid has a PEG moiety with an average molecular weight ranging from 1,000 g / mol to 5,000 g / mol.

27. The pharmaceutical composition of an antiviral agent for use according to claim 25, wherein the PEG-modified lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)] (DSPE-PEG).

28. 28. The pharmaceutical composition of an antiviral agent for use according to claim 27, wherein the one or more phospholipids are neutral phospholipids, and 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.

29. 25. The pharmaceutical composition of an antiviral agent for use according to claim 24, wherein the antiviral agent is an antimalarial agent or an antiretroviral agent.

30. the antiviral agent is a nucleoside compound of structural formula (I): 【Chemistry 2】 Each R １ , R ２ , R ３ , R ４ or R ５ are independently H, OR ａ , N(R ａ ) ２ , N ３ , C.N., N.O. ２ , S(O) ｎ R ａ , halogen or methyl, and n is 0, 1 or 2; R ６ is CN or H, Each R ａ are independently H, (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, aryl (C １ ~C ８ ) alkyl, (C ４ ~C ８ ) carbocyclylalkyl, —C(═O)R １１ , -C(=O)OR １１ , —C(═O)NR １１ R １２ , -C(=O)SR １１ , -S(O)R １１ , -S(O) ２ R １１ , -S(O)(OR １１ ), -S(O) ２ (OR １１ ), or -SO ２ NR １１ R １２ and R ７ is H, each X １ or X ２ are independently, C-R １０ or N, R ８ is halogen, NR １１ R １２ , N(R １１ ) (OR １１ ), N.R. １１ NR １１ R １２ , N ３ , NO, NO ２ , 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 , (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, aryl (C １ ~C ８ ) alkyl, (C ４ ~C ８ ) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)(C １ ~C ８ ) alkyl, —S(O) ｎ (C １ ~C ８ ) alkyl, aryl (C １ ~C ８ ) alkyl, OR １１ or SR １１ wherein each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Each R ９ or R １０ are independently H, halogen, R １１ , OR １１ , S.R. １１ , N.R. １１ R １２ , N(R １１ ) (OR １１ ), N.R. １１ NR １１ R １２ , N ３ , NO, NO ２ , 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 are independently H, (C １ ~C ８ ) alkyl, (C ２ ~C ８ ) alkenyl, (C ２ ~C ８ ) alkynyl, (C ４ ~C ８ ) carbocyclylalkyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)(C １ ~C ８ ) alkyl, —S(O) ｎ (C １ ~C ８ ) alkyl or aryl (C １ ~C ８ ) alkyl, and each aryl or heteroaryl is independently optionally substituted with one or more Z groups; Or, R １１ or R １２ together with the nitrogen to which they are both attached form a 3- to 7-membered heterocycle, and any one carbon atom of said heterocycle is optionally —O—, —S—, or —NR ａ - may be replaced by Each Z group independently represents 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, -NO ２ , = N ２ , -N ３ , -NHC(=O)R ｂ , -OC(=O)R ｂ , -NHC(=O)NR ｂ ２ , -S(=O) ２ -, -S(=O) ２ OH, -S(=O) ２ R ｂ , -OS(=O) ２ OR ｂ , -S(=O) ２ OH, -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 are independently H, alkyl, aryl, arylalkyl, or heterocycle, and each of the above (C １ ~C ８ ) One or more of the non-terminal carbon atoms of the alkyl is —O—, —S—, or —NR ａ may be substituted with - 25. An antiviral pharmaceutical composition for use according to claim 24.

31. The antiviral agent is 【Transformation 3】 【Chemistry 4】 25. The pharmaceutical composition of an antiviral agent for use according to claim 24, selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

32. 32. The pharmaceutical composition of an antiviral agent for use according to any one of claims 24 to 31, having a concentration of said antiviral agent in the range of 0.1 to 80 mg / mL.

33. 32. A pharmaceutical composition of an antiviral agent for use according to any one of claims 24 to 31, having a concentration of said antiviral agent in the range of 0.5 to 5 mg / mL.

34. 32. An aerosolized composition of particles containing a liposomal antiviral agent for use in treating an infectious disease, the aerosolized composition of particles comprising a plurality of particles of an antiviral pharmaceutical composition for use in treating a respiratory disease according to any one of claims 24 to 31.

35. 35. The aerosolized composition of particles of claim 34, wherein the plurality of particles have a mass median aerodynamic diameter in the range of about 0.5 μm to 5 μm.

36. 1. A method of treating an infectious or respiratory disease, comprising: Administering the antiviral pharmaceutical composition for use according to any one of claims 24 to 31 to a subject in need thereof. A method comprising:

37. 37. The method of claim 36, wherein the respiratory disease is selected from the group consisting of severe pneumonia, acute respiratory infection (SARI) including acute respiratory distress syndrome (ARDS), sepsis, and septic shock.

38. 1. A method for reducing complications associated with the treatment of a respiratory or infectious disease in a human subject, comprising: Administering the composition of any one of claims 1 to 16 to a subject in need thereof. A method comprising:

39. 39. The method of claim 38, wherein the complication comprises cardiac or hepatotoxicity.

40. 39. The method of claim 38, wherein the complication comprises prolongation of corrected QT interval (QTc).

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