Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
  • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
  • A61K9/1605—Excipients; Inactive ingredients
  • A61K9/1629—Organic macromolecular compounds
  • A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
  • A61K31/7088—Compounds having three or more nucleosides or nucleotides
  • A61K31/713—Double-stranded nucleic acids or oligonucleotides
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K9/00—Medicinal preparations characterised by special physical form
  • A61K9/0012—Galenical forms characterised by the site of application
  • A61K9/0043—Nose
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
  • A61P31/12—Antivirals
  • A61P31/14—Antivirals for RNA viruses
  • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
• • C—CHEMISTRY; METALLURGY
  • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
  • C08L—COMPOSITIONS OF MACROMOLECULAR COMPOUNDS
  • C08L3/00—Compositions of starch, amylose or amylopectin or of their derivatives or degradation products
  • C08L3/02—Starch; Degradation products thereof, e.g. dextrin

Definitions

• POLYINOSINIC-POLYCYTIDYLIC ACID PEA STARCH FORMULATION FOR THE PREVENTION AND/OR TREATMENT OF UPPER RESPIRATORY TRACT INFECTIONS
• the common cold (also known as nasopharyngitis, acute viral rhinopharyngitis, acute coryza, or a cold) is a viral infectious disease of the upper respiratory system caused primarily by viruses. Also influenza or influenza-like illnesses are caused by viral infections.
• the common cold is a viral infection of the upper respiratory tract.
• the most commonly implicated virus is the rhinovirus (30-50%), a type of picornavirus with 99 known serotypes.
• Others include coronavirus (10-15%), influenza (5-15%), human parainfluenza viruses, human respiratory syncytial virus, adenoviruses, enteroviruses, and metapneumovirus.
• coronaviruses are particularly implicated in adult colds. Of over 30 coronaviruses, 3 or 4 cause infections in humans, but they are difficult to grow in the laboratory and their significance is thus less well-understood. Due to the many different types of viruses and their tendency for continuous mutation, it is impossible to gain complete immunity to the common cold.
• the first indication of an upper respiratory virus is often a sore or scratchy throat.
• Other common symptoms are runny nose, congestion, and sneezing. These are sometimes accompanied by conjunctivitis (pink eye), muscle aches, fatigue, malaise, headache, weakness, or loss of appetite.
• Cough and fever generally indicate influenza rather than an upper respiratory virus with a positive predictive value of around 80%. Symptoms may be more severe in infants and young children, and in these cases it may include fever and hives. Upper respiratory viruses may also be more severe in smokers.
• Viral replication begins 2 to 6 hours after initial contact. Symptoms usually begin 2 to 5 days after initial infection but occasionally occur in as little as 10 hours. Symptoms peak 2-3 days after symptom onset, whereas influenza symptom is constant and immediate. There is currently no known treatment that shortens the duration; however, symptoms usually resolve spontaneously in 7 to 10 days, with some symptoms possibly lasting for up to three weeks. In children the cough lasts for more than 10 days in 35-40% and continues for more than 25 days in 10% of the cases. The common cold is the most frequent infectious disease in humans with the average adult contracting two to four infections a year and the average child contracting several infections per year between 6-12 years of age.
• the common cold is most infectious during the first two to three days of symptoms however it is also infectious for a couple of days before the onset of symptoms and may still be somewhat infectious until symptoms have completely resolved.
• HRV Human rhinovirus
• the HRV particle is comprised of a 27-30 nm non-enveloped capsid consisting of 4 polypeptides (VPl, VP2, VP3, and VP4).
• the virus capsid contains a single-stranded RNA genome of approximately 7200 bases.
• a virally-encoded protein (VPg) is covalently attached to the 5' end of the RNA genome.
• HRVs can infect the upper and lower airways, nasal mucosa, sinuses and middle ear, and infections produce symptoms of "the common cold" (see above). Infections are self-limiting and are typically restricted to the upper airways. Peripheral white blood cell counts may be elevated during the first 2-3 days of the infection.
• HRV infection can also lead to infection of the lower airways, otitis media
• Symptomatic treatment generally involves using sleep-inducing oral anti-histamines and /or vaso-constrictive decongestants that have stimulant side-effects. This is only marginally effective and these side-effects are often as debilitating as the infection itself. Although prevention would be the ideal solution, for the reasons cited above the chances of a broadly effective vaccine against all the different serotypes is highly unlikely in the near future. So, short of quarantine, people will be exposed to these infectious agents on a regular basis, especially during "cold season" and so a broadly effective, convenient, side- effect free prophylactic would have a major impact on public health and productivity in the work place.
• the present invention relates to a composition
• a composition comprising micro particles of polyinosinic-polycytidylic acid (Poly (I:C)) and a pea starch polymer for use in treating and/or preventing infections such as viral respiratory infections or the common cold and a device, preferably a nasal delivery system, comprising said composition for use by a patient in need to prevent and/or treat said infections or the common cold.
• Poly (I:C) polyinosinic-polycytidylic acid
• a pea starch polymer for use in treating and/or preventing infections such as viral respiratory infections or the common cold
• a device preferably a nasal delivery system
• Figure 1 consists of two panels, labeled panel (A) and panel (B). Each panel is a graph depicting the radiance of a region of interest (ROI) for mice administered
• ROI region of interest
• microparticles comprising polyinosinic-polycytidylic acid (PolylC) and pea starch (Lycoat RS780®) at a ratio of 1 :3 (PolylC/Lycoat RS780 1/3), 1 :5 (PolylC/Lycoat RS780 1/5), or 1 : 12 (PolylC/Lycoat RS780 1/12), as well as a negative control (Placebo starch Lycoat RS780).
• the mice comprised a firefly luciferase gene in the interferon beta (INF- ⁇ ) locus, and thus, radiance correlates with INF- ⁇ expression.
• Panel A and B depict the same data plotted with either a linear y-axis (A) or a logarithmic y-axis (B).
• Figure 2 consists of two panels, labeled panel (A) and panel (B). Each panel contains images of mice comprising a gene for firefly luciferase in the interferon beta (INF- ⁇ ) locus that were administered luciferin, either 2-3 hours prior to administering
• the top row of four images in panels (A) and (B) are replicates of mice that were administered microparticles comprising polyinosinic-polycytidylic acid (PolylC) and pea starch (Lycoat RS780®) at a ratio of 1 :3 (left mouse in each image), 1 :5 (center mouse in each image), or 1 : 12 (right mouse in each image).
• the middle row of four images depicts two control mice, which were each imaged separately (left image and right-center image), and two images of three mice each (left-center image and right image), which were ordered as in the top row.
• the bottom row of three images consists of mice that were administered microparticles comprising polyinosinic-polycytidylic acid (PolylC) and pea starch (Lycoat RS780®) at a ratio of 1 :3 (left image and center image, each depicting three mice) and mice that were administered microparticles comprising solely pea starch (Lycoat RS780®; right image, depicting three mice).
• Figure 3 consists of two panels, labeled panel (A) and panel (B). Each panel is a graph depicting the radiance of a region of interest (ROI) for mice administered
• ROI region of interest
• microparticles comprising polyinosinic-polycytidylic acid (PolylC) and pea starch (Lycoat RS780®) at a ratio of 1 :3 (PolylC/Lycoat RS780 1/3), 1 :5 (PolylC/Lycoat RS780 1/5), or 1 : 12 (PolylC/Lycoat RS780 1/12), as well as a negative control (Placebo starch Lycoat RS780).
• the mice comprised a firefly luciferase gene in the interferon beta (INF- ⁇ ) locus, and thus, radiance correlates with INF- ⁇ expression.
• Panel A and B depict the same data plotted with either a linear y-axis (A) or a logarithmic y-axis (B).
• Figure 3 is based on a second experiment designed to determine whether the results of Figure 1 are reproducible.
• Figure 4 consists of two panels, labeled panel (A) and panel (B). Each panel is a graph depicting the radiance of a region of interest (ROI) for mice administered
• ROI region of interest
• microparticles comprising polyinosinic-polycytidylic acid (PolylC) and pea starch (Lycoat RS780®) at a ratio of 1 :3 (PolylC/Lycoat RS780 1/3), 1 :5 (PolylC/Lycoat RS780 1/5), or 1 : 12 (PolylC/Lycoat RS780 1/12), as well as a negative control (Placebo starch Lycoat RS780).
• the mice comprised a firefly luciferase gene in the interferon beta (INF- ⁇ ) locus, and thus, radiance correlates with INF- ⁇ expression.
• Panel A and B depict the same data plotted with either a linear y-axis (A) or a logarithmic y-axis (B).
• Figure 4 is based on pooled data, including the data points in Figures 1 and 3 as well as a third experiment.
• a formulation of a triggering molecule (Poly (I:C)) that can be used in a measurable and controllable fashion, for example, every couple of days, once a week or even every few weeks to prime the innate immune system and provide protection against viral infection.
• Poly (I:C) triggering molecule
• the approach outlined below takes an existing agent, Poly (I:C), which has demonstrated efficacy, but which is impractical and renders it convenient and effective using formulation sciences.
• TLR3 Toll-like receptor 3
• TLR3 is a protein that in humans is encoded by the TLR3 gene.
• TLR3 is a member of the Toll-like receptor family of pattern recognition receptors of the innate immune system which plays a fundamental role in pathogen recognition and activation of innate immunity.
• TLRs are highly conserved from Drosophila to humans and share structural and functional similarities. They recognize pathogen-associated molecular patterns (PAMPs) that are expressed on infectious agents, and mediate the production of cytokines necessary for the development of effective immunity.
• PAMPs pathogen-associated molecular patterns
• the various TLRs exhibit different patterns of expression.
• This TLR3 receptor is also expressed by airway epithelial cells and is restricted to the dendritic subpopulation of the leukocytes.
• TLR3 recognizes double-stranded RNA (dsRNA). Double-stranded RNA is RNA with two complementary strands that can be formed during the viral replication cycle. Upon recognition, TLR 3 induces the activation of transcription factors like F- ⁇ and Interferon Regulatory Factor 3 (IRF3) to increase production of type I interferon which signal other cells to increase their antiviral defenses.
• the structure of TLR3 forms a large horseshoe shape that contacts with a neighboring horseshoe, forming a "dimer" of two horseshoes. Much of the TLR3 protein surface is covered with sugar molecules, making it a glycoprotein, but on one face
• This surface also contains two distinct patches rich in positively-charged amino acids, which may be a binding site for negatively-charged double-stranded RNA.
• Poly (I:C) Polyinosine-polycytidylic acid
• Poly (I:C) is a double stranded RNA molecule with a MW distribution up to, for instance 3.600.000 Daltons.
• Poly (I:C) is a Toll Like Receptor 3 (TLR3) ligand that mimics viral RNA and is a known stimulant of the innate immune response. When administered nasally, it induces expression of anti-viral proteins like interferon a and ⁇ in the nasal epithelium. It has been demonstrated to reduce the number and severity of rhinovirus infections.
• TLR3 Toll Like Receptor 3
• Poly (I:C) is usually an unstable molecule in normal aqueous solutions.
• Poly (I:C) needs to be re- dissolved immediately prior to use and administered every 2 hours.
• a novel formulation has been developed that is stable and shows enhanced efficacy.
• Poly (I:C) has now been formulated in the present invention with a bio-adhesive polymer, pea starch, that can prolong the residence time on the nasal epithelium and provide a more effective and controllable stimulation of the innate immune system.
• the current invention surprisingly provides the identification of a unique formulation that could be stored almost indefinitely at room temperature and which retains its innate immune system-stimulating activity.
• the current inventive formulation contains water-soluble pea starch and has the advantage of low viscosity characteristics.
• composition comprising Poly (I:C)
• such composition shows surprisingly a less sticky behavior to the inside of a vial, tube or device (like sprays) when such vials, tubes or devices must be filled with the inventive composition compared to any other starch used for the same purpose.
• the technical advantage is that more precisely the dosage of said Poly (I:C) can be administered to the patient in need, since less composition will stick to the inner side of the nasal spray device accordingly.
• the formulation enhances the efficacy of Poly (I:C) and permits much less frequent dosing with even greater TLR3 stimulating activity.
• the invention therefore relates to a composition
• a composition comprising micro particles of polyinosinic-polycytidylic acid (Poly (I:C)) and a pea starch polymer in the ratio Poly (I:C) over pea starch of 1 : 3.
• Poly (I:C) polyinosinic-polycytidylic acid
• pea starch polymer in the ratio Poly (I:C) over pea starch of 1 : 3.
• Micro particles obtained and used in the current invention are particles with an average particle size between 0.1 ⁇ and 100 ⁇ .
• the carrier polymer is starch obtained from the plant genus Lathyrus, more specifically from peas.
• Poly (LC)-carrier-polymer microspheres, or also so-called micro particles, comprised in the composition are produced by means of a particle formation process such as a spray-dry process.
• composition of the invention can be a bi-phasic suspension or a liquid composition comprising an organic solvent, wherein the organic solvent is based on glycerol or ethanol or a combination thereof.
• organic solvents serving the same purpose are trifluranes or other etherous propellants.
• composition of the invention can be used in human and/or animal medicine preferably for use in preventing and/or treating viral infections of the human upper respiratory tract such as what are referred to as "common colds.”
• composition according to the invention can be used as aerosol formulation in for instance stables, barns, chicken flocks, and the like.
• the current composition can be used by patients suffering from asthma and/or COPD (Chronic Obstructive Pulmonary Disease) in order to potentially prevent and/or treat upcoming common cold symptoms or illnesses and thus prevent exacerbations of their underlying illnesses / symptoms.
• COPD Cho Obstructive Pulmonary Disease
• composition of the current invention comprising micro particles of
• polyinosinic-polycytidylic acid Poly (I:C)
• pea starch polymer can be used for the treatment and/or prevention of (viral) infections or common cold, wherein the composition is administered by nasal application at a time interval that is in the range of one day to one month, more preferably from every couple of days or even once a week or bi-weekly.
• composition wherein the ratio Poly (I:C) / pea starch is 1/3 in combination with the micro particle size in the composition ranging from 0.1 ⁇ to 200 ⁇ , preferably from 1 ⁇ to 50 ⁇ , more preferably from 2 ⁇ to 40 ⁇ , even more preferably from 2 ⁇ to 20 ⁇ , and most preferred from 10 ⁇ to 20 ⁇ , highly preferable is 13, 14 or 15 ⁇ , can be used for the treatment and/or prevention of (viral) infections or common cold, wherein said composition is administered by nasal application at a time interval that is in the range of one day to one month, more preferably from every couple of days or even once a week or bi-weekly.
• Part of the invention is also a device, in particular a nasal delivery system, comprising a composition according to the invention.
• Poly (I:C) is formulated as a dry powder for nasal administration.
• Poly (I:C) is spray dried from an aqueous mixture containing pea starch and Poly (I:C).
• Starch is believed to have a dual function: (1) to act as a bio-adhesive in the nose, (2) and to serve as protective matrix for stabilizing Poly (I:C).
• Starch, especially pea starch, is a preferred excipient for nasal application as accumulation is prevented by degradation through amylases.
• Starches with high amylopectin content or with chemically modified starches exhibit good muco-adhesion.
• hydroxypropylated (pregelatinized) pea starch (chemically modified) is used in the present invention, as these are cold water-swelling and contain a cold water-soluble fraction, resulting in a homogeneous dispersion when mixed at low shear with Poly (I:C).
• the resulting starch dispersions have a low to medium viscosity which allows spray drying into a homogeneous powder even at high starch ratios.
• Nasal administration is preferably achieved using a single dose nasal powder device (Unit dose device supplied from Aptar Pharma Germany).
• the unit dose device is an active delivery system, meaning that the patient does not need to inhale and performance is patient independent. Dosing is performed by actuation, which is controlled by overpressure. The dose per puff is determined by the concentration of Poly (I:C) in the spray dried powder and the emitted weight of the powder. The powder will be administered into each nostril using a new device for each puff.
• Poly (I:C) is a synthetic double-stranded RNA composed of anti-parallel polynucleotide strands of inosinic acid and cytidylic acid sodium salts.
• the strands are non-covalently bound by hydrogen bonds formed between the inosine and cytosine bases.
• the average chain length for the Poly (I:C) ranges between 300 to 6,000 base pairs, corresponding to approximately 180,000 to about 3,600,000 daltons.
• the molecular formula is CioHi 0 N 4 Na0 7 P)x ⁇ (C 9 HnNaN 3 0 7 P) x .
• Poly (I:C) can be purchased, but it can optionally also be made in house using, for instance, the following procedure.
• the duplex product Poly (I:C) is manufactured from the individual homopolymers Poly Inosine (I) and Poly Cytidine (C).
• Poly I and Poly C are synthesized by individually polymerizing the nucleoside diphosphates inosine and cytidine in the presence of polynucleotide phosphorylase (P Pase). Each nucleoside diphosphate is individually polymerized by PNPase for 20-24 hours to control the length of the resulting ribonucleic acid polymer.
• the enzyme, protein kinase is then added to terminate the polymerization reaction.
• the resulting homopolymers are hydrolyzed to control the molecular weight range of each polymer product within a specified range.
• the hydrolyzed product is treated with ethanol to precipitate the single stranded RNA molecules (ssRNA) from solution.
• the precipitate is separated from the supernatant and dissolved in water.
• the solution of ssRNA is then filtered to remove particulates, ultra filtered to remove the low- molecular weight contaminants, and then lyophilized. Lyophilized ssRNA products are individually tested for purity, molecular weight, and other quality attributes to ensure the products are within specification.
• the individual single stranded homo-polymers (Poly I and Poly C) are individually dissolved in 0.015 M sodium chloride and then combined to anneal the strands forming the double stranded duplex product (Poly I : Poly C). After mixing, the resulting solution is filtered. The filtrate is ultra-filtered to remove low molecular weight contaminants. The ultra-filtered product is then lyophilized. The resulting duplex product is stored at -20 °C. The lyophilized dsRNA product is tested for purity, molecular weight, and other quality attributes to ensure the product is within specification.
• the invention relates to a composition
• a composition comprising micro particles of polyinosinic-polycytidylic acid (Poly (I:C)) and a pea starch in a ratio 1/3 (w/w).
• the Poly (I:C) - pea starch micro particles may be produced, for example, by means of a particle formation process such as a spray-dry process.
• the D v 50 of the micro particle may range from 0.1 ⁇ to 200 ⁇ .
• the composition may be a liquid composition comprising an organic solvent, e.g., based on glycerol or ethanol or a combination thereof.
• the invention provides a device comprising the composition, such as a nasal delivery device.
• the composition may be used in medicine, e.g., in treating and/or preventing infections, common cold or influenza-like illnesses.
• the composition may be used for the manufacture of a medicament for preventing and/or treating upper respiratory infections by nasal administration.
• the invention relates to a microparticle, comprising polyinosinic- polycytidylic acid and a starch.
• the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio greater than 1 :9.
• the microparticle may comprise polyinosinic-polycytidylic acid and starch at a ratio of about 1 : 1 to about 1 :8, such as about 1 : 1 to about 1 :7, about 1 : 1 to about 1 :6, about 1 : 1 to about 1 :5, about 1 : 1 to about 1 :4, about 1 : 1 to about 1 :3, about 1 :2 to about 1 :8, about 1 :2 to about 1 :7, about 1 :2 to about 1 :6, about 1 :2 to about 1 :5, about 1 :2 to about 1 :4, about 1 :2 to about 1 :3, about 1 :3 to about 1 :8, about 1 :3 to about 1 :7, about 1 :3 to about 1 :6, about 1 :3 to about 1 :5, or about
• the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio of about 1 : 1 to about 1 :7. In more preferable embodiments, the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio of about 1 : 1 to about 1 :6. In even more preferable embodiments, the microparticle comprises polyinosinic- polycytidylic acid and starch at a ratio of about 1 :2 to about 1 :5. In the most preferable embodiments, the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio of about 1 :3 to about 1 :5.
• the microparticle may comprise polyinosinic-polycytidylic acid and starch at a ratio of about 1 : 1, about 1 :2, about 1 :3, about 1 :4, about 1 :5, about 1 :6, about 1 :7, or about 1 :8.
• the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio of about 1 : 1, about 1 :2, about 1 :3, about 1 :4, about 1 :5, about 1 :6, or about 1 :7.
• the microparticle comprises polyinosinic- polycytidylic acid and starch at a ratio of about 1 : 1, about 1 :2, about 1 :3, about 1 :4, about 1 :5, or about 1 :6. In even more preferable embodiments, the microparticle comprises polyinosinic-polycytidylic acid and starch at a ratio of about 1 :2, about 1 :3, about 1 :4, or about 1 :5. In the most preferable embodiments, the microparticle comprises polyinosinic- polycytidylic acid and starch at a ratio of about 1 :3, about 1 :4, or about 1 :5.
• the starch may be maize starch (i.e., cornstarch), wheat starch, potato starch, or pea starch.
• the starch may be banana starch, rice starch, barley starch, rye starch, millet starch, oat starch, yam starch, sweet potato starch, cassava starch (i.e., tapioca starch), sago starch, arrowroot starch, fava bean starch, lentil starch, mung bean starch, or chickpea starch.
• the starch may comprise starch from more than one source, e.g., the starch may comprise pea starch, potato starch, wheat starch, and/or maize starch.
• the starch comprises a pea starch, or even consists or consists essentially of a pea starch.
• the starch comprises a maize starch (e.g., waxy maize starch), or even consists essentially of a maize starch (e.g., waxy maize starch).
• the starch may comprise at least about 20% amylose, such as at least about 25% amylose or at least about 30% amylose. In certain embodiments, the starch comprises at least about 25% amylose.
• the starch may comprise about 20% amylose to about 85% amylose, such as about 20% to about 40%, about 30% to about 50%, about 40% to about 60%, about 50% to about 70%, about 60% to about 80%, about 20% to about 30%, about 25% to about 35%, about 30% to about 40%, about 35% to about 45%, about 40% to about 50%, about 45% to about 55%, about 50% to about 60%, about 55% to about 65%, about 60% to about 70%, about 65% to about 75%, about 70% to about 80%, or about 75% to about 85%) amylose.
• the starch may comprise 0% to about 10% amylose, such as 0% to about 5%, 0% to about 4%, 0% to about 3%, 0% to about 2%, or 0% to about 1% amylose.
• the starch may comprise a high amylose starch, such as high amylose maize starch (e.g., EURYLON®).
• the starch comprises about 15% amylose to about 50% amylose, more preferably about 20% amylose to about 45% amylose, and most preferably about 25% amylose to about 40% amylose.
• a high amylose starch such as high amylose maize starch (e.g., EURYLON®).
• the starch may comprise less than about 5% amylose, such as less than 4%, less than about 3%, less than about 2%, or less than about 1% amylose.
• the fraction of the starch that is not amylose is preferably amylopectin, e.g., a starch that comprises about 20% to about 40%) amylose preferably comprises about 60% to about 80% amylopectin and a starch that comprises 0% to about 10% amylose preferably comprises about 90% to 100% amylopectin.
• the starch may comprise about 15% to about 80% amylopectin, such as about 20% to about 40%, about 30% to about 50%, about 40% to about 60%, about 50% to about 70%, about 60% to about 80%, about 15% to about 25%, about 20% to about 30%, about 25% to about 35%, about 30% to about 40%, about 35% to about 45%, about 40% to about 50%, about 45% to about 55%, about 50% to about 60%, about 55% to about 65%, about 60% to about 70%), about 65% to about 75%, or about 70% to about 80% amylopectin.
• the starch may comprise about 90% to 100% amylopectin, such as about 95% to 100%), about 96% to 100%, about 97% to 100%, or about 98% to 100% amylopectin.
• the starch comprises about 50% to about 85% amylopectin, more preferably about 55%) to about 80% amylopectin, and most preferably about 60% to about 75% amylopectin. In some preferred embodiments, the starch comprises at least about 90% amylopectin, such as at least about 95%, about 96%, about 97%, about 98%, or at least about 99%) amylopectin.
• the fraction of the starch that is not amylopectin is preferably amylose, e.g., a starch that comprises about 60% to about 70% amylopectin preferably comprises about 30% to about 40% amylose and a starch that comprises about 95% to 100%) amylopectin preferably comprises 0% to about 5% amylose.
• the starch is a pregelatinized starch.
• the starch may be pregelatinized pea starch or pregelatinized maize starch (e.g., pregelatinized waxy maize starch). Starch gelatinization may increase the solubility of the starch.
• the starch may be a modified starch, such as dextrin, acid-treated starch, alkaline- treated starch, bleached starch, oxidized starch, enzyme-treated starch, monostarch phosphate, distarch phosphate, phosphated distarch phosphate, acetylated distarch phosphate, starch acetate, acetylated distarch adipate, hydroxypropyl starch, hydroxypropyl distarch phosphate, hydroxypropyl distarch glycerol, starch sodium octenyl succinate, starch aluminum octenyl succinate, acetylated oxidized starch, cationic starch, hydroxyethyl starch, or carboxymethylated starch.
• a modified starch such as dextrin, acid-treated starch, alkaline- treated starch, bleached starch, oxidized starch, enzyme-treated starch, monostarch phosphate,
• Lycoat RS780® is a pregelatinized hydroxypropyl pea starch, wherein some of the hydroxyl groups of the pea starch have been substituted with a hydroxypropyl group. In some embodiments, 0% to about 40% of the hydroxyl groups of the starch are
• the starch is a hydroxypropyl starch, such as a hydroxypropyl pea starch.
• the starch is a pregelatinized hydroxypropyl starch, such as a pregelatinized hydroxypropyl pea starch.
• the starch is a hydroxyethyl starch, such as hydroxyethyl maize starch.
• the starch is a pregelatinized hydroxyethyl starch, such as a pregelatinized hydroxyethyl maize starch.
• the average chain length of the polyinosinic-polycytidylic acid is approximately 300 base pairs to 6,000 base pairs.
• the polyinosinic- polycytidylic acid may comprise polyinosinic acid having an average chain length of approximately 300 nucleotides to 6,000 nucleotides and polyinosinic acid having an average chain length of approximately 300 nucleotides to 6,000 nucleotides.
• the average molecular weight of the polyinosinic-polycytidylic acid is approximately 180 kDa to 3,600 kDa.
• the polyinosinic-polycytidylic acid may be present as a salt, such as a salt of sodium.
• the microparticle may comprise water.
• a microparticle may consist essentially of polyinosinic-polycytidylic acid, starch, and water, e.g., wherein polyinosinic-polycytidylic acid comprises sodium or another cation.
• a microparticle may consist of polyinosinic-polycytidylic acid, starch, and water.
• the microparticle may have a size from 0.1 ⁇ to 200 ⁇ , preferably from 1 ⁇ to 50 ⁇ , more preferably from 2 ⁇ to 40 ⁇ , even more preferably from 2 ⁇ to 20 ⁇ , and most preferred from 10 ⁇ to 20 ⁇ .
• the microparticle may have a size of about 2 ⁇ to about 30 ⁇ , such as about 4 ⁇ to about 30 ⁇ , about 5 ⁇ to about 30 ⁇ , or about 6 ⁇ to about 30 ⁇ .
• the microparticle may have a size of about 2 ⁇ to about 27 ⁇ , such as about 4 ⁇ to about 27 ⁇ , about 5 ⁇ to about 27 ⁇ , or about 6 ⁇ to about 27 ⁇ .
• the microparticle may have a size of about 2 ⁇ to about 20 ⁇ , such as about 4 ⁇ to about 20 ⁇ , about 5 ⁇ to about 20 ⁇ , or about 6 ⁇ to about 20 ⁇ m.
• the microparticle may have a size of about 2 ⁇ to about 10 ⁇ , such as about 4 ⁇ to about 10 ⁇ , about 5 ⁇ to about 10 ⁇ , or about 6 ⁇ to about 10 ⁇ .
• the invention relates to a composition comprising a plurality of microparticles as described herein (e.g., supra).
• the composition may have a D v 50 of about 1 ⁇ to about 200 ⁇ , such as about 1 ⁇ to about 50 ⁇ , about 2 ⁇ to about 40 ⁇ , about 2 ⁇ to about 20 ⁇ , or about 10 ⁇ to about 20 ⁇ .
• the composition may have a D v 50 of about 4 ⁇ to about 20 ⁇ , about 5 ⁇ to about 20 ⁇ , about 6 ⁇ to about 20 ⁇ , about 7 ⁇ m to about 20 ⁇ , about 8 ⁇ to about 20 ⁇ , about 9 ⁇ m to about 20 ⁇ , or about 10 ⁇ to about 20 ⁇ .
• a composition comprising a plurality of microparticles may be a liquid or a dry powder.
• a composition comprising a plurality of microparticles may comprise an organic solvent such as glycerol, ethanol, or a combination thereof.
• a composition comprising a plurality of microparticles may further comprise phosphate-buffered saline.
• a composition comprising a plurality of microparticles is stable during storage at room temperature for at least 1 month, such as at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, or at least 12 months.
• a composition comprising a plurality of microparticles may be stable during storage at room temperature for about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, or about 12 months.
• the invention relates to a nasal delivery system, comprising a composition comprising microparticles as described herein (e.g., supra).
• the invention relates to the use of a composition comprising a plurality of microparticles as described herein for the manufacture of a medicament for prophylacting against and/or treating upper respiratory infections. In some embodiments, the invention relates to the use of a composition comprising microparticles as described herein for the manufacture of a medicament for prophylacting against upper respiratory viral infections in a subject having chronic obstructive pulmonary disease (COPD).
• COPD chronic obstructive pulmonary disease
• a composition comprising a plurality of microparticles as described herein may be useful for treating or prophylacting against a viral infection of the upper respiratory tract, such as a rhinovirus infection or an influenza virus infection.
• the viral infection may be caused by a picornavirus (e.g., rhinovirus), coronavirus, influenza virus, human
• a composition comprising a plurality of microparticles as described herein may be useful for prophylacting against upper respiratory viral infections in a subject having chronic obstructive pulmonary disease (COPD), or a symptom thereof.
• a method may comprise administering the composition to a subject, e.g., by nasal administration.
• the subject may be a human or an animal, preferably human.
• the invention relates to a method of treating or prophylacting against a viral infection in a subject, comprising administering to the subject a composition comprising a plurality of microparticles as described herein.
• the viral infection may be caused by a picornavirus (e.g., rhinovirus), coronavirus, influenza virus, human
• Administering a composition may comprise, for example, nasal administration.
• the subject may be a human or an animal, preferably human.
• the invention relates to a method of prophylacting against upper respiratory viral infections in a subject having chronic obstructive pulmonary disease
• composition comprising a plurality of microparticles as described herein.
• Administering a composition may comprise, for example, nasal administration.
• the subject may be a human or an animal, preferably human.
• Example 1 Methods of preparing and characterizing microparticles
• the solutions were fed to a two-fluid nozzle (diameter: 0.7 mm) at the top of the spray dryer by means of a peristaltic pump.
• the spray dryer operated in co-current nitrogen flow mode.
• the spray dried particles were collected in a reservoir attached to a cyclone. After collection of the particles, the glass cylinder and cyclone was cooled to room temperature.
• the collected powder was transferred to amber glass bottle and this bottle is placed in an aluminum vapor lock bag.
• the vials were stored at room temperature.
• the samples were sputtered with gold particles with diameter +/- 30-50 nm. Images were generated using a FEI scanning electron microscope -type Quanta 200F with Everhart Thornley detector.
• Water content of the concepts was determined by means of a direct volumetric Karl Fisher titration.
• a KF TITRATOR V30 is used (Mettler Toledo, US).
• the powder 50-100 mg was transferred to the titration vessel containing Hydranal® Methanol Dry (Sigma Aldrich) and stirred for 300 seconds.
• Titration was performed with Hydranal® Composite 2 (Sigma Aldrich) at a concentration of 2 mg/ml using a 5 ml burette. For termination, a stop drift of 15 ⁇ g/min was applied. Samples were analyzed in triplicate.
• the instrument is used in the blue light ON detection mode at a size range of 20 nm to 2 mm.
• the measured particle size distribution by volume in the current invention for D v 10 is 4 ⁇ , for D v 50 it is 14 ⁇ while for D v 90 it is 27 ⁇ .
• Example 2 in vivo testing of formulations in the influenza mouse model
• the luciferase reporter for interferon-beta (IFN- ⁇ ) gene activation can provide insight in the activation of IFN- ⁇ after stimulation with different poly IC formulations.
• Poly IC is a synthetic analog which mimics dsRNA viruses by stimulation of the innate immune system through Pattern Recognition Receptors (PRR).
• PRR Pattern Recognition Receptors
• TLR3 PRR
• RIG-1 and/or MDA 5 a signaling cascade is started and results in activation of type I interferons, of which IFN- ⁇ is a representative.
• the heterozygous IFN-P+/Ap-luc albino (Tyrc2J) C57BL/6 mice produce firefly luciferase driven by the IFN- ⁇ promoter due to a targeted mutation in the IFN- ⁇ locus.
• luciferin is administered systemically and photon emission is monitored using an IVIS200 system (CaliperLS).
• the compounds were administered intranasally to IFN- ⁇ reporter mice and in vivo imaging was performed before and 24h after administration.
• mice The generation of the IFN- ⁇ reporter mice has been previously described
• mice produce firefly luciferase driven by the IFN- ⁇ promoter due to a targeted mutation in the IFN- ⁇ locus.
• the mice used in this study were heterozygous IFN-P+/Ap-luc albino (Tyrc2J) C57BL/6. Males and females between the age of 12 and 14 weeks were used. The animals are housed in IVC racks and they are provided with food and water ad libitum.
• mice Thirty one 8-12 week old male and female heterozygous IFN"P+/Ap-luc albino (Tyrc2J) C57BL/6 were used. All intranasal administrations were performed under injection anesthesia (ketamine/xylazine). The dry powder was administered using a self-made tip device. The tip-devices were used to administer 2 mm of powder into the left nostril and an additional 2 mm of powder onto the nose. After the administration of the powder, the mice were placed under a red lamp to wake up.
• Figure 2 depicts mice imaged for luciferase activity before ( Figure 2A) and after ( Figure 2B) administering the microparticles.
• Figure 3 depicts the measured radiance of the mice after administering microparticles comprising either a 1 :3, 1 :5, or 1 : 12 polyinosinic-polycytidylic acid to starch (Lycoat RS780®) ratio, or the negative control (Placebo starch Lycoat RS780).

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