US20180235879A1 - Inhalable powder formulations of alginate oligomers - Google Patents

Inhalable powder formulations of alginate oligomers Download PDF

Info

Publication number
US20180235879A1
US20180235879A1 US15/507,712 US201515507712A US2018235879A1 US 20180235879 A1 US20180235879 A1 US 20180235879A1 US 201515507712 A US201515507712 A US 201515507712A US 2018235879 A1 US2018235879 A1 US 2018235879A1
Authority
US
United States
Prior art keywords
particles
spray dried
dried particles
alginate oligomer
phospholipid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US15/507,712
Other languages
English (en)
Inventor
Bildad NYAMBURA
Anand BAKLE
Arne Dessen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Algipharma AS
Original Assignee
Algipharma AS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Algipharma AS filed Critical Algipharma AS
Assigned to ALGIPHARMA AS reassignment ALGIPHARMA AS ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: BAKLE, Anand, NYAMBURA, Bildad, DESSEN, ARNE
Publication of US20180235879A1 publication Critical patent/US20180235879A1/en
Abandoned legal-status Critical Current

Links

Images

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
    • 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
    • A61K9/007—Pulmonary tract; Aromatherapy
    • A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • A61K9/0075—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a dry powder inhaler [DPI], e.g. comprising micronized drug mixed with lactose carrier particles
    • 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
    • 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
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • 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
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • A61K31/734—Alginic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • 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
    • A61K9/007—Pulmonary tract; Aromatherapy
    • A61K9/0073—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy
    • A61K9/008—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy comprising drug dissolved or suspended in liquid propellant for inhalation via a pressurized metered dose inhaler [MDI]
    • 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
    • A61K9/10—Dispersions; Emulsions
    • 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
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • 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
    • A61K9/1617—Organic compounds, e.g. phospholipids, fats
    • A61K9/1623—Sugars or sugar alcohols, e.g. lactose; Derivatives thereof; Homeopathic globules
    • 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
    • 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
    • A61K9/1682—Processes
    • 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
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/4808—Preparations in capsules, e.g. of gelatin, of chocolate characterised by the form of the capsule or the structure of the filling; Capsules containing small tablets; Capsules with outer layer for immediate drug release
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/0028—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M15/00—Inhalators
    • A61M15/0028—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up
    • A61M15/003—Inhalators using prepacked dosages, one for each application, e.g. capsules to be perforated or broken-up using capsules, e.g. to be perforated or broken-up
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system

Definitions

  • the present invention relates to novel formulations of alginate oligomers that are especially suited for inhalation routes of administration. More specifically, the present invention provides spray dried particles for inhalation in dry powder form containing a high proportion of alginate oligomer, a phospholipid and an anti-adherent compound.
  • the uncomplicated formulation of the invention provides particles with a fine particle mass (FPM), geometric particle size distribution and emitted dose that permits effective delivery of the alginate oligomer to the lungs of a subject undergoing treatment with minimised risk of adverse reaction to excipients.
  • the invention further provides dry powder compositions for inhalation containing the spray dried particles, capsules containing said particles and dry powder inhalers containing said particles.
  • the invention further provides a surprising and advantageous method for preparing the particles of the invention in which an organic-in-aqueous liquid emulsion of the alginate oligomer, the phospholipid and the anti-adherent compound is prepared and said emulsion is spray dried.
  • the use of the particles of the invention in therapy, in particular the treatment or prevention of respiratory infections and respiratory disorders is provided, especially disorders or conditions which involve microbial infection, in particular biofilm infection, and/or abnormal mucus, e.g.
  • COPD COPD
  • COAD COAD
  • COAP bronchitis
  • cystic fibrosis a medical disorder or condition associated with CF
  • emphysema a medical disorder or condition associated with CF
  • lung cancer emphysema
  • asthma or pneumonia or more broadly any condition associated with or characterised by defective cystic fibrosis transmembrane conductance regulator (CFTR) ion channel function.
  • CFTR cystic fibrosis transmembrane conductance regulator
  • alginates are linear polymers of (1-4) linked ⁇ -D-mannuronic acid (M) and/or its C-5 epimer ⁇ -L-guluronic acid (G).
  • M linked ⁇ -D-mannuronic acid
  • G C-5 epimer
  • the primary structure of alginates can vary greatly.
  • the M and G residues can be organised as homopolymeric blocks of contiguous M or G residues, as blocks of alternating M and G residues and single M or G residues can be found interspacing these block structures.
  • An alginate molecule can comprise some or all of these structures and such structures might not be uniformly distributed throughout the polymer.
  • Alginate oligomers may be obtained from alginate polymers which are typically isolated from natural sources as large high molecular weight polymers (e.g. an average molecular weight in the range 300,000 to 500,000 Daltons). Such large alginate polymers may be degraded, or broken down, e.g. by chemical or enzymatic hydrolysis to produce alginate structures of lower molecular weight.
  • Alginate oligomers have been shown to be able to reduce the viscosity of mucus, in particular hyperviscous mucus, to have antimicrobial properties and also antibiofilm properties.
  • Their use in the treatment or prevention of respiratory infections and respiratory disorders in which microbial infection, in particular biofilm infection and/or abnormal mucus are involved, e.g. COPD, COAD, COAP, bronchitis, cystic fibrosis, a medical disorder or condition associated with CF, emphysema, lung cancer, asthma and pneumonia have therefore been suggested, amongst others.
  • alginate oligomers cannot be successfully formulated as pure dry powders with a fine particle mass (FPM), geometric particle size distribution and emitted dose that permit effective delivery of the alginate oligomer to the lungs. It has however surprisingly been found that when the alginate oligomer is combined with relatively small amounts of a phospholipid and an anti-adherent compound, particles can be prepared by spray drying which have a fine particle mass (FPM), geometric particle size distribution and emitted dose that permit effective delivery of the alginate oligomer to the lungs.
  • FPM fine particle mass
  • the surfactant properties of the phospholipid are believed to enable a partial masking of the alginate oligomer in the spray dried powder, reducing the hydrophilicity and thereby the hygroscopic nature of the powder. This is further believed to lead to the desired size distribution and to reduce deposition in the mouth and trachea rather than the lungs, thereby increasing bioavailability in the lungs.
  • the particles of the invention contain at least about 71% w/w of the alginate oligomer, e.g. at least about 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89 or 90% w/w of the alginate oligomer. In other preferred embodiments the particles of the invention contain about 70% to about 90% w/w of the alginate oligomer, e.g.
  • the particles of the invention contain about 80% w/w of the alginate oligomer, e.g. 78, 79, 80, 81 or 82% w/w, preferably 79, 80 or 81% w/w, more preferably 80% w/w.
  • the particles of the invention contain at least about 11% w/w in total of the phospholipid and the anti-adherent compound, e.g. at least about 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30% w/w in total of the phospholipid and the anti-adherent compound. In other preferred embodiments the particles of the invention contain about 10% to about 30% w/w in total of the phospholipid and the anti-adherent compound, e.g. any one of about 11, 12, 13, 14, 15, 16, 17, 18, 19, 20% w/w to any one of about 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30% w/w in total of the phospholipid and the anti-adherent compound, e.g.
  • the particles of the invention contain about 20% w/w in total of the phospholipid and the anti-adherent compound, e.g. 18, 19, 20, 21 or 22% w/w, preferably 19, 20 or 21% w/w, more preferably 20% w/w.
  • the particles of the invention contain no less than 0.5% w/w of the phospholipid, e.g. no less than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 25 or 29.5% w/w. In preferred embodiments the particles of the invention contain 0.5-15% w/w of the phospholipid, e.g.
  • the particles of the invention contain about 5% w/w of the phospholipid, e.g. 3, 4, 5, 6 or 7% w/w, preferably 4, 5 or 6% w/w, more preferably 5% w/w.
  • the particles of the invention contain no less than 0.5% w/w of the anti-adherent compound, e.g. no less than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 25 or 29.5% w/w. In preferred embodiments the particles of the invention contain than 0.5-20% w/w of the anti-adherent compound, e.g.
  • the particles of the invention contain about 15% w/w of the anti-adherent compound, e.g. 13, 14, 15, 16 or 17% w/w, preferably 14, 15 or 16% w/w, more preferably 15% w/w.
  • the particles of the invention contain about 5% w/w of the phospholipid, e.g. 3, 4, 5, 6 or 7% w/w, preferably 4, 5 or 6% w/w, more preferably 5% w/w and about 15% w/w of the anti-adherent compound, e.g. 13, 14, 15, 16 or 17% w/w, preferably 14, 15 or 16% w/w, more preferably 15% w/w. More preferably the particles of the invention contain about 5% w/w of the phospholipid and about 15% w/w of the anti-adherent compound.
  • the relative amounts of phospholipid and anti-adherent compound present in the particles are in a ratio of 1:5 to 5:1, preferably 1:4.5 to 4.5:1, 1:4 to 4:1, 1:3.5 to 3.5:1, 1:3 to 3:1, 1:2.5 to 2.5:1, 1:2 to 2:1, 1:1.5 to 1.5:1 or 1:1, more preferably 1:3.5, 1:3 or 1:2.5, most preferably 1:3.
  • the relative amounts of the alginate oligomer and the combined amounts of phospholipid and anti-adherent compound present in the particles are in a ratio of 7:3, 7:2.5, 7:2, 7.5:2.5, 7.5:1.5, 8:2, 8:1.5, 8:1, 8.5:1.5, 8.5:1 or 9:1.
  • the relative amounts of the alginate oligomer, the phospholipid and anti-adherent compound present in the particles are in a ratio of 7:2:1, 7:1.5:1.5, 7:1:2, 7:0.5:2.5, 7:0.05:2.95, 8:1.5:0.5, 8:1:1, 8:0.5:1.5, 8:0.05:1.95, 9:0.75:0.25, 9:0.5:0.5, 9:0.25:0.75, or 9:0.5:0.95, 8:0.5:1.5 is preferred.
  • no further excipients are present
  • the particles of the invention contain no greater than about 9% w/w of further excipients, e.g. no greater than about 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1, 0.05, 0.01, 0.005 or 0.001% w/w of further excipients. In most preferred embodiments the particles of the invention contain essentially no further excipients. Suitable further excipients are disclosed herein. In certain embodiments a further excipient is not a phospholipid, e.g. those which are solid at room temperature, or an anti-adherent compound.
  • the particles of the invention contain about 80% w/w alginate oligomer, about 15% w/w anti-adherent compound and about 5% w/w phospholipid and essentially no further excipients.
  • the sum of the individual percentage amounts of each component will be 100%. It may be that upon preparation the particles of the invention retain solvent molecules, e.g. water molecules, in intimate association with the molecules of alginate oligomer, phospholipid, anti-adherent compound and/or further excipient. For the purposes of the present invention residual solvent associated in this way is included pro rata in the % w/w calculations and so has no impact on relative amounts of the components of particles of the invention.
  • solvent molecules e.g. water molecules
  • % w/w (or “percentage weight by weight”) is a commonly used expression designating the proportion of a solid composition that is made up of the compound in question. 1% w/w equates to 1 gram of compound per 100 grams of solid composition, 2% w/w equates to 2 g of compound per 100 g of solid composition, and so on. 1% w/w also equates to 10 gram of compound per kilogram of solid composition.
  • the particles of the invention permit effective delivery of the alginate oligomer to the lungs of a subject undergoing treatment.
  • the particles of the invention are of a size appropriate, or suitable, for inhalation, that is for delivery to the lungs of a subject by means of a dry powder inhaler (i.e. an inhaler device suitable for administration of a powder).
  • a dry powder inhaler i.e. an inhaler device suitable for administration of a powder.
  • the particles have an aerodynamic particle size of less than 5 ⁇ m, for example up to 4.9, 4.8, 4.7, 4.6, or 4.5 ⁇ m.
  • the particles have a mass median aerodynamic diameter (MMAD) of less than 5 ⁇ m, e.g. up to 4.9, 4.8, 4.7, 4.6, or 4.5 ⁇ m.
  • MMAD mass median aerodynamic diameter
  • the particles advantageously have a uniform, or substantially uniform, size distribution, i.e.
  • the particles have a relatively narrow aerodynamic particle size distribution (APSD).
  • APSD may be defined with reference to the fine particle mass (FPM), as defined further below.
  • FPM fine particle mass
  • the particles are readily aerosolizable at relatively low aerodynamic dispersion forces.
  • the particles of the invention preferably have a geometric particle size distribution that is acceptable for inhalation, i.e. a d50 of ⁇ 5 ⁇ m and a d90 of ⁇ 10 ⁇ m and preferably a d10 of ⁇ 1.5 ⁇ m as measured in accordance with the protocol of the Examples, i.e. as measured using a Malvern Mastersizer MicroPlus particle size analyer (Malvern Instruments, UK) following dispersion of 15 mg particles in 3 ml of ethyl acetate containing sorbitane trioleate (SPAN 85) at a concentration of 2 g/l.
  • a geometric particle size distribution that is acceptable for inhalation, i.e. a d50 of ⁇ 5 ⁇ m and a d90 of ⁇ 10 ⁇ m and preferably a d10 of ⁇ 1.5 ⁇ m as measured in accordance with the protocol of the Examples, i.e. as measured using a Malvern Mastersizer MicroPlus particle size ana
  • the particles of the invention may a d50 of ⁇ 4 ⁇ m or ⁇ 3 or ⁇ 2.5 ⁇ m. In other embodiments the particles of the invention may have a d50 of about or approximately 3 ⁇ m, e.g. a d50 of 2-4, 2.5-4.0, 2.5-3.8, 2.5-3.5, 2.8-3.8, 2.8-3.5, or 2.8-3.2 ⁇ m.
  • the particles of the invention may a d90 of ⁇ 8 ⁇ m or ⁇ 7 or ⁇ 6 ⁇ m.
  • the particles of the invention may a d10 of ⁇ 2 ⁇ m or ⁇ 1 or ⁇ 0.5 ⁇ m.
  • any and all combinations of d10, d50 and/or d90 are contemplated, although in certain embodiments the particles may be described by reference to only a single d parameter, e.g. a d50 of ⁇ 5 ⁇ m or a d50 of ⁇ 3 ⁇ m. In another embodiment the particles of the invention may have a d50 of ⁇ 3 ⁇ m.
  • the particles of the invention preferably have a FPM (size distribution ⁇ 4.46 ⁇ m) of greater than about 10 mg, e.g. 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg per 40 mg of particles as measured in accordance with the protocol of the Examples, i.e.
  • Plastiape monodose device as the particle delivery means. This may be expressed as a percentage FPM ⁇ 4.46 ⁇ m of greater than about 25%, e.g. 28%, 30%, 33%, 35%, 38%, 40%, 43%, 45%, 48%, or 50%.
  • the particles of the invention preferably have an emitted dose of greater than about 65%, e.g. 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79 or 80% as measured in accordance with the protocol of the Examples, i.e. as determined using a next generation impactor (NGI) and a high resistance (60 L) Plastiape monodose device as the particle delivery means.
  • NTI next generation impactor
  • 60 L high resistance
  • the particles of the invention preferably display at least about 90%, e.g. at least 91, 92, 93, 94, 95, 96, 97, 98 or 99% release of alginate oligomer upon exposure of 4 mg of particles to at least 0.1 ml saline for at least one hour at room temperature.
  • alginates typically occur as polymers of an average molecular mass of at least 35,000 Daltons, i.e. approximately 175 to approximately 190 monomer residues, although typically much higher and an alginate oligomer according to the present invention may be defined as a material obtained by fractionation (i.e. size reduction) of an alginate polymer, commonly a naturally occurring alginate.
  • An alginate oligomer can be considered to be an alginate of an average molecular weight of less than 35,000 Daltons (i.e. less than approximately 190 or less than approximately 175 monomer residues), in particular an alginate of an average molecular weight of less than 30,000 Daltons (i.e. less than approximately 175 or less than approximately 150 monomer residues) more particularly an average molecular weight of less than 25,000 or 20,000 Daltons (i.e. less than approximately 135 or 125 monomer residues or less than approximately 110 or 100 monomer residues).
  • an oligomer generally comprises 2 or more units or residues and an alginate oligomer for use according to the invention will typically contain 2 to 100 monomer residues, more typically 3, 4, 5 or 6 to 100, and may contain 2, 3, 4, 5 or 6 to 75, 2, 3, 4, 5 or 6 to 50, 2, 3, 4, 5 or 6 to 40, 2, 3, 4, 5 or 6 to 35 or 2, 3, 4, 5 or 6 to 30 residues.
  • an alginate oligomer for use according to the invention will typically have an average molecular weight of 350, 550, 700, 900 or 1000 to 20,000 Daltons, 350, 550, 700, 900 or 1000 to 15,000 Daltons, 350, 550, 700, 900 or 1000 to 10,000 Daltons, 350, 550, 700, 900 or 1000 to 8000 Daltons, 350, 550, 700, 900 or 1000 to 7000 Daltons, or 350, 550, 700, 900 or 1000 to 6,000 Daltons.
  • the alginate oligomer may have a degree of polymerisation (DP), or a number average degree of polymerisation (DPn) of 2 to 100, preferably 2 to 75, preferably 2 to 50, more preferably 2 to 40, 2 to 35, 2 to 30, 2 to 28, 2 to 25, 2 to 22, 2 to 20, 2 to 18, 2 to 17, 2 to 15 or 2 to 12.
  • DP degree of polymerisation
  • DPn number average degree of polymerisation
  • An alginate oligomer will, as noted above, contain (or comprise) guluronate or guluronic acid (G) and/or mannuronate or mannuronic acid (M) residues or units.
  • An alginate oligomer according to the invention will preferably be composed solely, or substantially solely (i.e. consist essentially of) uronate/uronic acid residues, more particularly solely or substantially solely of G and/or M residues.
  • at least 80%, more particularly at least 85, 90, 95 or 99% of the monomer residues may be uronate/uronic acid residues, or, more particularly G and/or M residues.
  • the alginate oligomer will not comprise other residues or units (e.g. other saccharide residues, or more particularly other uronic acid/uronate residues).
  • the alginate oligomer is preferably a linear oligomer.
  • At least 30% of the monomer residues of the alginate oligomer are G residues (i.e. guluronate or guluronic acid).
  • the alginate oligomer will contain at least 30% guluronate (or guluronic acid) residues.
  • Specific embodiments thus include alginate oligomers with (e.g. containing) 30 to 70% G (guluronate) residues or 70 to 100% G (guluronate) residues.
  • a representative alginate oligomer for use according to the present invention may contain at least 70% G residues (i.e. at least 70% of the monomer residues of the alginate oligomer will be G residues).
  • the alginate oligomer may be an oligoguluronate (i.e. a homooligomer of G, or 100% G)
  • the above described alginates of the invention have a primary structure wherein the majority of the G residues are in so called G-blocks.
  • G-blocks Preferably at least 50%, more preferably at least 70 or 75%, and most preferably at least 80, 85, 90, 92 or 95% of the G residues are in G-blocks.
  • a G block is a contiguous sequence of at least two G residues, preferably at least 3 contiguous G residues, more preferably at least 4 or 5 contiguous G residues, most preferably at least 7 contiguous G residues.
  • At least 90% of the G residues are linked 1-4 to another G residue. More particularly at least 95%, more preferably at least 98%, and most preferably at least 99% of the G residues of the alginate are linked 1-4 to another G residue.
  • the alginate oligomer of use in the invention is preferably a 3- to 35-mer, more preferably a 3- to 28-mer, in particular a 4- to 25-mer, e.g. a 5- to 20-mer, especially a 6- to 22-mer, in particular an 8- to 20-mer, especially a 10- to 15-mer, e.g. having a molecular weight in the range 350 to 6400 Daltons or 350 to 6000 Daltons, preferably 550 to 5500 Daltons, preferably 750 to 5000 Daltons, and especially 750 to 4500 Daltons or 2000 to 3000 Daltons or 900 to 3500 Daltons.
  • Other representative alginate oligomers include, as mentioned above, oligomers with 5, 6, 7, 8, 9, 10, 11, 12 or 13 to 50, 45, 40, 35, 28, 25, 22 or 20 residues.
  • the monomeric residues in the alginate oligomer may be the same or different and not all need carry electrically charged groups although it is preferred that the majority (e.g. at least 60%, preferably at least 80% more preferably at least 90%) do. It is preferred that a substantial majority, e.g. at least 80%, more preferably at least 90% of the charged groups have the same polarity.
  • the ratio of hydroxyl groups to charged groups is preferably at least 2:1, more especially at least 3:1.
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 3-28, 4-25, 6-22, 8-20 or 10-15, or 5-18 or 7-15 or 8-12, especially 10.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 3-24, 4-23, 5-22, 6-21, 7-20, 8-19, 9-18, 10-17, 11-16, 12-15 or 13-14 (e.g. 13 or 14).
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DPn), of 4-25, 5-24, 6-23, 7-22, 8-21, 9-20, 10-19, 11-18, 12-17, 13-16, 14-15 (e.g. 14 or 15).
  • DP degree of polymerisation
  • DPn number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 5-26, 6-25, 7-24, 8-23, 9-22, 10-21, 11-20, 12-19, 13-18, 14-17 or 15-16 (e.g. 15 or 16).
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 4-50, 4-40, 4-35, 4-30, 4-28, 4-26, 4-22, 4-20, 4-18, 4-16 or 4-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 5-50, 5-40, 5-25, 5-22, 5-20, 5-18, 5-23, 5-20, 5-18, 5-16 or 5-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 6-50, 6-40, 6-35, 6-30, 6-28, 6-26, 6-24, 6-20, 6-19, 6-18, 6-16 or 6-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 8-50, 8-40, 8-35, 8-30, 8-28, 8-25, 8-22, 8-20, 8-18, 8-16 or 8-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 9-50, 9-40, 9-35, 9-30, 9-28, 9-25, 9-22, 9-20, 9-18, 9-16 or 9-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 10-50, 10-40, 10-35, 10-30, 10-28, 10-25, 10-22, 10-20, 10-18, 10-16 or 10-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 11-50, 11-40, 11-35, 11-30, 11-28, 11-25, 11-22, 11-20, 11-18, 11-16 or 11-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 12-50, 12-40, 12-35, 12-30, 12-28, 12-25, 12-22, 12-20, 12-18, 12-16 or 12-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 13-50, 13-40, 13-35, 13-30, 13-28, 13-25, 13-22, 13-20, 13-18, 13-16 or 13-14.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 14-50, 14-40, 14-35, 14-30, 14-28, 14-25, 14-22, 14-20, 14-18, 14-16 or 14-15.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 15-50, 15-40, 15-35, 15-30, 15-28, 15-25, 15-22, 15-20, 15-18 or 15-16.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention may have a degree of polymerisation (DP), or a number average degree of polymerisation (DP n ), of 18-50, 18-40, 18-35, 18-30, 18-28, 18-25, 18-22 or 18-20.
  • DP degree of polymerisation
  • DP n number average degree of polymerisation
  • the alginate oligomer of the invention is substantially free, preferably essentially free, of alginate oligomers having a degree of polymerisation outside of the ranges disclosed herein.
  • This may be expressed in terms of the molecular weight distribution of the alginate oligomer of the invention, e.g. the percentage of each mole of the alginate oligomer being used in accordance with the invention which has a DP outside the relevant range.
  • the molecular weight distribution is preferably such that no more than 10%, preferably no more than 9, 8, 7, 6, 5, 4, 3, 2, or 1% mole has a DP of three, two or one higher than the relevant upper limit for DP n .
  • no more than 10%, preferably no more than 9, 8, 7, 6, 5, 4, 3, 2, or 1% mole has a DP below a number three, two or one smaller than the relevant lower limit for DP n .
  • Suitable alginate oligomers are described in WO2007/039754, WO2007/039760, WO 2008/125828, and WO2009/068841, the disclosures of which are explicitly incorporated by reference herein in their entirety.
  • Representative suitable alginate oligomers have a DP n in the range 5 to 30, a guluronate fraction (F G ) of at least 0.80, a mannuronate fraction (F M ) of no more than 0.20, and at least 95 mole % of DP no more than 25.
  • alginate oligomers have a number average degree of polymerization in the range 7 to 15 (preferably 8 to 12), a guluronate fraction (F G ) of at least 0.85 (preferably at least 0.90), a mannuronate fraction (F M ) of no more than 0.15 (preferably no more than 0.10), and having at least 95% mole with a degree of polymerization less than 17 (preferably less than 14).
  • alginate oligomers have a number average degree of polymerization in the range 5 to 18 (especially 7 to 15), a guluronate fraction (F G ) of at least 0.80 (preferably at least 0.85, especially at least 0.92), a mannuronate fraction (F M ) of no more than 0.20 (preferably no more than 0.15, especially no more than 0.08), and having at least 95% mole with a degree of polymerization less than 20 (preferably less than 17).
  • alginate oligomers have a number average degree of polymerization in the range 5 to 18, a guluronate fraction (F G ) of at least 0.92, a mannuronate fraction (F M ) of no more than 0.08, and having at least 95% mole with a degree of polymerization less than 20.
  • Suitable alginate oligomers have a number average degree of polymerization in the range 5 to 18 (preferably 7 to 15, more preferably 8 to 12, especially about 10), a guluronate fraction (F G ) of at least 0.80 (preferably at least 0.85, more preferably at least 0.90, especially at least 0.92, most especially at least 0.95), a mannuronate fraction (F M ) of no more than 0.20 (preferably no more than 0.15, more preferably no more than 0.10, especially no more than 0.08, most especially no more than 0.05), and having at least 95% mole with a degree of polymerization less than 20 (preferably less than 17, more preferably less than 14).
  • alginate oligomers have a number average degree of polymerization in the range 7 to 15 (preferably 8 to 12), a guluronate fraction (F G ) of at least 0.92 (preferably at least 0.95), a mannuronate fraction (F M ) of no more than 0.08 (preferably no more than 0.05), and having at least 95% mole with a degree of polymerization less than 17 (preferably less than 14).
  • alginate oligomers have a number average degree of polymerization in the range 5 to 18, a guluronate fraction (F G ) of at least 0.80, a mannuronate fraction (F M ) of no more than 0.20, and having at least 95% mole with a degree of polymerization less than 20.
  • alginate oligomers have a number average degree of polymerization in the range 7 to 15, a guluronate fraction (F G ) of at least 0.85, a mannuronate fraction (F M ) of no more than 0.15, and having at least 95% mole with a degree of polymerization less than 17.
  • alginate oligomers have a number average degree of polymerization in the range 7 to 15, a guluronate fraction (F G ) of at least 0.92, a mannuronate fraction (F M ) of no more than 0.08, and having at least 95% mole with a degree of polymerization less than 17.
  • alginate oligomers have a number average degree of polymerization in the range 5 to 20, a guluronate fraction (F G ) of at least 0.85 and a mannuronate fraction (F M ) of no more than 0.15.
  • alginate oligomers have a number average degree of polymerization in the range 5 to 20, a guluronate fraction (F G ) of 0.9-0.95 and a mannuronate fraction (F M ) of 0.05-0.1, which may be expressed as an alginate oligomer having 90-95% G residues and an average molecular weight of 2600 Da.
  • Further suitable alginate oligomers have a number average degree of polymerization about 13 (e.g. 12, 13 or 14), a guluronate fraction (F G ) of at least about 0.80, 0.85, 0.87, 0.88, 0.90 or 0.93 (e.g. 0.92, 0.93 or 0.94) and a corresponding mannuronate fraction (F M ) of no more than about 0.20, 0.15, 0.13, 0.12, 0.10, or 0.07 (e.g. 0.08, 0.07 or 0.06).
  • alginate oligomers have a number average degree of polymerization about 21 (e.g. 20, 21 or 22), a guluronate fraction (F G ) of at least about 0.80 (e.g. 0.85, 0.87, 0.88, 0.90, 0.92, 0.94 or 0.95) and a corresponding mannuronate fraction (F M ) of no more than about 0.20 (e.g. 0.15, 0.13, 0.12, 0.10, 0.08, 0.06, 0.05).
  • F G guluronate fraction
  • F M corresponding mannuronate fraction
  • alginate oligomers have a number average degree of polymerization about 6 (e.g. 5, 6 or 7), a guluronate fraction (F G ) of at least about 0.80 (e.g. 0.85, 0.87, 0.88, 0.90, 0.92, 0.94 or 0.95) and a corresponding mannuronate fraction (F M ) of no more than about 0.20 (e.g. 0.15, 0.13, 0.12, 0.10, 0.08, 0.06, 0.05).
  • F G guluronate fraction
  • F M corresponding mannuronate fraction
  • alginate oligomers favoured according to the present invention is alginate oligomers defined as so-called “high G” or “G-block” oligomers i.e. having a high content of G residues or G-blocks (e.g. wherein at least 70% of the monomer residues are G, preferably arranged in G-blocks).
  • high G or “G-block” oligomers
  • G-block oligomers
  • alginate oligomers with high proportions of a single monomer type, and with said monomers of this type being present predominantly in contiguous sequences of that monomer type, that represent oligomers that are particularly preferred, e.g. oligomers wherein at least 70% of the monomer residues in the oligomer are G residues linked 1-4 to another G-residue, or more preferably at least 75%, and most preferably at least 80, 85, 90, 92, 93, 94, 95, 96, 97, 98, 99% of the monomers residues of the oligomer are G residues linked 1-4 to another G residue.
  • This 1-4 linkage of two G residues can be alternatively expressed as a guluronic unit bound to an adjacent guluronic unit.
  • At least, or more particularly more than, 50% of the monomer residues of the alginate oligomer may be M residues (i.e. mannuronate or mannuronic acid).
  • the alginate oligomer will contain at least or alternatively more than 50% mannuronate (or mannuronic acid) residues.
  • Specific embodiments thus include alginate oligomers with (e.g. containing) 50 to 70% M (mannuronate) residues or e.g. 70 to 100% M (mannuronate) residues.
  • Further specific embodiments also include oligomers containing 71 to 85% M residues or 85 to 100% M residues.
  • a representative alginate oligomer for use according to this embodiment of the present invention will contain more than 70% M residues (i.e. more than 70% of the monomer residues of the alginate oligomer will be M residues).
  • the alginate oligomer may be an oligomannuronate (i.e. a homooligomer of M, or 100% M).
  • the above described alginates of the invention have a primary structure wherein the majority of the M residues are in so called M-blocks.
  • M-blocks preferably at least 50%, more preferably at least 70 or 75%, and most preferably at least 80, 85, 90 or 95% of the M residues are in M-blocks.
  • An M block is a contiguous sequence of at least two M residues, preferably at least 3 contiguous M residues, more preferably at least 4 or 5 contiguous M residues, most preferably at least 7 contiguous M residues.
  • At least 90% of the M residues are linked 1-4 to another M residue. More particularly at least 95%, more preferably at least 98%, and most preferably at least 99% of the M residues of the alginate are linked 1-4 to another M residue.
  • oligomers are alginate oligomers wherein at least 70% of the monomer residues in the oligomer are M residues linked 1-4 to another M-residue, or more preferably at least 75%, and most preferably at least 80, 85, 90, 92, 93, 94, 95, 96, 97, 98, 99% of the monomers residues of the oligomer are M residues linked 1-4 to another M residue.
  • This 1-4 linkage of two M residues can be alternatively expressed as a mannuronic unit bound to an adjacent mannuronic unit.
  • the alginate oligomers of the invention comprise a sequence of alternating M and G residues.
  • a sequence of at least three, preferably at least four, alternating M and G residues represents an MG block.
  • the alginate oligomers of the invention comprise an MG block.
  • an MG block is a sequence of at least three contiguous residues consisting of G and M residues and wherein each non-terminal (internal) G residue in the contiguous sequence is linked 1-4 and 4-1 to an M residue and each non-terminal (internal) M residue in the contiguous sequence is linked 1-4 and 4-1 to a G residue.
  • the MG block is at least 5 or 6 contiguous residues, more preferably at least 7 or 8 contiguous residues.
  • the minority uronate in the alginate oligomer is found predominantly in MG blocks.
  • the alginate oligomer is arranged such that at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, e.g. 100% of the G and M residues in the oligomer are arranged in MG blocks.
  • the invention extends to embodiments wherein at least 1% but less than 100% of the monomer residues of the oligomer are G residues (i.e. guluronate or guluronic acid), more particularly, and as defined further below, at least 30% of the monomer residues are G residues.
  • the MG block containing alginate oligomer may contain at least 1%, but less than 100%, guluronate (or guluronic acid) residues, but generally the MG block containing alginate oligomer will contain at least 30% (or at least 35, 40 or 45% or 50% G) but less than 100% G.
  • Specific embodiments thus include MG block containing alginate oligomers with (e.g.
  • a representative MG block containing alginate oligomer for use according to the present invention may contain more than 30%, but less than 70%, G residues (i.e. more than 30%, but less than 70%, of the monomer residues of the MG block alginate oligomer will be G residues).
  • the MG block containing alginate oligomer can have e.g. between 35% and 65%, 40% and 60% or 45% and 55% G residues.
  • the MG block containing alginate oligomer may have approximately equal amounts of G and M residues (e.g. ratios between 65% G/35% M and 35% G/65% M, for instance 60% G/40% M and 40% G/60% M; 55% G/45% M and 45% G/55% M; 53% G/47% M and 47% G/53% M; 51% G/49% M and 49% G/51% M; e.g. about 50% G and about 50% M) and these residues are arranged predominantly, preferably entirely or as completely as possible, in an alternating MG pattern (e.g. at least 50% or at least 60, 70, 80, 85, 90 or 95% or 100% of the M and G residues are in an alternating MG sequence).
  • G and M residues e.g. ratios between 65% G/35% M and 35% G/65% M, for instance 60% G/40% M and 40% G/60% M; 55% G/45% M and 45% G/55% M; 53% G/47% M and 47% G/53% M; 51%
  • the terminal uronic acid residues of the oligomers of the invention do not have a double bond, especially a double bond situated between the C 4 and C 5 atom.
  • Such oligomers may be described as having saturated terminal uronic acid residues. The skilled man would be able to prepare oligomers with saturated terminal uronic acid residues without undue burden. This may be through the use of production techniques which yield such oligomers, or by converting (saturating) oligomers produced by processes that yield oligomers with unsaturated terminal uronic acid residues.
  • the alginate oligomer will typically carry a charge and so counter ions for the alginate oligomer may be any physiologically tolerable ion, especially those commonly used for charged drug substances, e.g. sodium, potassium, ammonium, chloride, mesylate, meglumine, etc. Ions which promote alginate gelation e.g. group 2 metal ions may also be used.
  • alginate oligomer may be a synthetic material generated from the polymerisation of appropriate numbers of guluronate and mannuronate residues
  • the alginate oligomers of use in the invention may conveniently be obtained, produced or derived from natural sources such as those mentioned above, namely natural alginate source materials.
  • Polysaccharide to oligosaccharide cleavage to produce the alginate oligomer useable according to the present invention may be performed using conventional polysaccharide lysis techniques such as enzymatic digestion and acid hydrolysis.
  • acid hydrolysis is used to prepare the alginate oligomers on the invention.
  • enzymatic digestion is used with an additional processing step(s) to saturate the terminal uronic acids in the oligomers.
  • Oligomers may then be separated from the polysaccharide breakdown products chromatographically using an ion exchange resin or by fractionated precipitation or solubilisation or filtration.
  • U.S. Pat. No. 6,121,441 and WO 2008/125828 which are explicitly incorporated by reference herein in their entirety, describe a process suitable for preparing the alginate oligomers of use in the invention. Further information and discussion can be found in for example in “Handbooks of Hydrocolloids”, Ed. Phillips and Williams, CRC, Boca Raton, Fla., USA, 2000, which textbook is explicitly incorporated by reference herein in its entirety.
  • the alginate oligomers may also be chemically modified, including but not limited to modification to add charged groups (such as carboxylated or carboxymethylated glycans) and alginate oligomers modified to alter flexibility (e.g. by periodate oxidation).
  • Alginate oligomers for example oligoguluronic acids
  • suitable for use according to the invention may conveniently be produced by acid hydrolysis of alginic acid from, but not limited to, Laminaria hyperbora and Lessonia nigrescens, dissolution at neutral pH, addition of mineral acid reduce the pH to 3.4 to precipitate the alginate oligomer (oligoguluronic acid), washing with weak acid, resuspension at neutral pH and freeze drying.
  • alginates for production of alginate oligomers of the invention can also be obtained directly from suitable bacterial sources e.g. Pseudomonas aeruginosa or Azotobacter vinelandii.
  • algal sources are expected to be most suitable on account of the fact that the alginates produced in these organisms tend to have these structures.
  • the bacterial sources may be more suitable for obtaining alginate oligomers of different structures.
  • the G content of alginates can be increased by epimerisation, for example with mannuronan C-5 epimerases from A. vinelandii or other epimerase enzymes.
  • epimerisation for example in vitro epimerisation may be carried out with isolated epimerases from Pseudomonas or Azotobacter, e.g. AlgG from Pseudomonas fluorescens or Azotobacter vinelandii or the AlgE enzymes (AlgE1 to AlgE7) from Azotobacter vinelandii.
  • AlgG from Pseudomonas or Azotobacter
  • AlgE enzymes AlgE1 to AlgE7 from Azotobacter vinelandii.
  • AlgE4 epimerase can be used to create alginates or alginate oligomers with alternating stretches of M/G sequence or primary structures containing single G residue as it has been found that this enzyme seems preferentially to epimerise individual M residues so as to produce single G residues linked to M residues rather than producing G blocks.
  • Particular primary structures can be obtained by using different combinations of these enzymes.
  • WO 94/09124 describes recombinant or modified mannuronan C-5 epimerase enzymes (AlgE enzymes) for example encoded by epimerase sequences in which the DNA sequences encoding the different domains or modules of the epimerases have been shuffled or deleted and recombined.
  • AlgE enzymes recombinant or modified mannuronan C-5 epimerase enzymes
  • mutants of naturally occurring epimerase enzymes, (AlgG or AlgE) may be used, obtained for example by site directed or random mutagenesis of the AlgG or AlgE genes.
  • a different approach is to create Pseudomonas and Azotobacter organisms that are mutated in some or all of their epimerase genes in such a way that those mutants produce alginates of the required structure for subsequent alginate oligomer production, or even alginate oligomers of the required structure and size (or molecular weight).
  • the generation of a number of Pseudomonas fluorescens organisms with mutated AlgG genes is described in detail in WO 2004/011628 and Gimmestad, M., et al, 2003 (supra).
  • the generation of a number of Azotobacter vinelandii organisms with mutated AlgE genes is disclosed in Gimmestad, M., et al, 2006 (supra).
  • a further approach is to delete or inactivate the endogenous epimerase genes from an Azotobacter or a Pseudomonas organism and then to introduce one or more exogenous epimerase genes, which may or may not be mutated (i.e. may be wild-type or modified) and the expression of which may be controlled, for example by the use of inducible or other “controllable promoters”.
  • exogenous epimerase genes which may or may not be mutated (i.e. may be wild-type or modified) and the expression of which may be controlled, for example by the use of inducible or other “controllable promoters”.
  • a still further approach would be to introduce some or all of the alginate biosynthesis machinery of Pseudomonas and/or Azotobacter into a non-alginate producing organism (e.g. E. coli ) and to induce the production of alginate from these genetically modified organisms.
  • a non-alginate producing organism e.g. E. coli
  • the primary structure of the alginate or alginate oligomer products can be influenced by the culture conditions. It is well within the capabilities of the skilled man to adjust culture parameters such as temperature, osmolarity, nutrient levels/sources and atmospheric parameters in order to manipulate the primary structure of the alginates produced by a particular organism.
  • references to “G residues/G” and “M residues/M” or to guluronic acid or mannuronic acid, or guluronate or mannuronate are to be read interchangeably as references to guluronic acid/guluronate and mannuronic acid/mannuronate (specifically ⁇ -L-guluronic acid/guluronate and ⁇ -D-mannuronic acid/mannuronate), and further include derivatives thereof in which one or more available side chains or groups have been modified without resulting in a capacity to treat or prevent a respiratory infection or a respiratory disorder, especially disorders or conditions which involve microbial infection, in particular biofilm infection, and/or abnormal mucus, e.g.
  • Common saccharide modifying groups would include acetyl, sulphate, amino, deoxy, alcohol, aldehyde, ketone, ester and anhydro groups.
  • the alginate oligomers may also be chemically modified to add charged groups (such as carboxylated or carboxymethylated glycans), and to alter flexibility (e.g. by periodate oxidation). The skilled man would be aware of still further chemical modifications that can be made to the monosaccharide subunits of oligosaccharides and these can be applied to the alginate oligomers of the invention.
  • the invention encompasses the use of a single alginate oligomer or a mixture (multiplicity/plurality) of different alginate oligomers.
  • a combination of different alginate oligomers e.g. two or more may be used.
  • the phospholipids of use in the invention are those which, when in a pure form, are solid at room temperature (about 20° C.) and at standard atmospheric pressure (1 atm; approximately 101325 Pa).
  • Representative examples include, but are not limited to, phosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerols, phosphatidylserines, phosphatidylinositols and combinations thereof, e.g.
  • phosphatidyl choline saturated and unsaturated
  • phosphatidyl ethanol amine phosphatidyl glycerol, phosphatidyl serine, phosphatidyl inositol, dioleoylphosphatidylcholine, dimyristoyl phosphatidylcholine , dipalmitoylphosphatidylcholine (DPPC; 1,2-Dipalmitoyl-snglycero-3-phosphocholine), distearoyl phosphatidylcholine, 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), diarachidoyl phosphatidylcholine, dibenoyl phosphatidylcholine, ditricosanoyl phosphatidylcholine, dilignoceroylphatidylcholine, dimiristoylphosphatidylethanolamine, dipalmitoyl-phosphatid
  • anti-adherent compound is used herein in its usual sense in the field of DPI formulations, i.e. a compound which combats adherence between particles thus preventing particle agglomeration. It will be readily understood however that the anti-adherent compound of the particles of the invention is not a phospholipid as described herein.
  • the anti-adherent compound may be an amino acid, e.g. a hydrophobic amino acid, in any isomeric form e.g. L- or D-. Amino acids will be well tolerated by subjects undergoing treatment in accordance with the invention.
  • Representative examples include but are not limited to histidine, alanine, isoleucine, arginine, leucine, asparagine, lysine, aspartic acid, methionine, cysteine, phenylalanine, glutamic acid, threonine, glutamine, tryptophan, glycine, valine, ornithine, proline, selenocysteine, serine, and tyrosine, e.g. glycine, lysine, proline, alanine, cysteine, methionine, valine, leucine, tyrosine, isoleucine, phenylalanine, tryptophan.
  • Preferred amino acids include leucine, isoleucine, alanine, valine, phenylalanine, lysine and glycine, e.g. leucine, lysine and glycine, in particular glycine.
  • Combinations of amino acids can also be employed, e.g. glycine and leucine.
  • combinations of hydrophobic and hydrophilic (preferentially partitioning in water) amino acids, where the overall combination is hydrophobic can also be employed.
  • Hydrophobicity is generally defined with respect to the partition of an amino acid between a nonpolar solvent and water.
  • Hydrophobic amino acids are those acids which show a preference for the nonpolar solvent. Relative hydrophobicity of amino acids can be expressed on a hydrophobicity scale on which glycine has the value 0.5. On such a scale, amino acids which have a preference for water have values below 0.5 and those that have a preference for nonpolar solvents have a value above 0.5.
  • the term “hydrophobic amino acid” refers to an amino acid that, on the hydrophobicity scale has a value greater than or equal to 0.5, in other words, has a tendency to partition in a nonpolar solvent which is at least equal to that of glycine.
  • anti-adherent compounds include mono- and disaccahrides.
  • the monosaccharide or one or more of the monosaccharide residues of the disaccharide may be a triose, a tetrose, a pentose, a hexose, a heptose, an octose, a nonose or a decose in pyranose or furanose form and/or L- or D-form where appropriate and/or sugar derivatives thereof.
  • Pentose or hexose saccharides/residues are preferred, e.g. mannose (e.g. D-mannose), galactose (e.g.
  • D-galactose glucose (e.g. D-glucose), fructose, fucose (e.g. L-fucose), N-acetyl-glucosamine, N-acetylgalactosamine, rhamnose, galactosamine, glucosamine (e.g.
  • D-glucosamine D-glucosamine
  • galacturonic acid glucuronic acid
  • N-acetylneuraminic acid N-acetylneuraminic acid
  • methyl D-mannopyranoside mannoside
  • a-methyl-glucoside galactoside
  • ribose xylose
  • arabinose saccharate
  • mannitol sorbitol
  • inositol glycerol and derivatives of these monomers.
  • the disaccharide may be exemplified by acarviosin, allolactose, cellobiose, chitobiose, galactose-alpha-1,3-galactose, dentiobiose, isomalt, isomaltose, isomaltulose, kojibiose, lactitol, lactobionic acid, lactose, lactulose, laminaribiose, maltitol, maltose, mannobiose, melibiose, melibiulose, neohesperidose, nigerose, robinose, rutinose, sambubiose, sophorose, sucralfate, sucralose, sucrose, sucrose acetate isobutyrate, sucrose octaacetate, trehalose, truranose, xylobiose or derivatives of these disaccharides.
  • the further excipient may be any pharmaceutically acceptable compound which may be included or formulated in a dry powder for inhalation, in particular for delivery to the lungs, which is not an alginate oligomer, a phospholipid or an anti-adherent compound as defined herein. Many such compounds are known in the art and selecting suitable compounds to meet precise requirements would be routine for the skilled person.
  • the further excipient maybe selected from starches, gum acacia, calcium phosphate, tragacanth, gelatine, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, propylene glycol, methyl cellulose, methylhydroxybenzoates, propyl hydroxybenzoates, talc, magnesium stearate, or suitable mixtures thereof. Additionally wetting agents, emulsifying agents, suspending agents, colouring agents, preserving agents, sweetening agents, flavouring agents, and the like may be used. As previously explained, the further excipient of the particles of the invention is not considered to encompass any residual solvent which may be retained following the preparation of the particles.
  • the particles are provided in the form of a dry powder consisting essentially of said particles.
  • the particles of the invention may be provided as a part of a dry powder composition with other dry powders.
  • Such other powders may contain one or more active agents, e.g. an antibiotic, an antifungal, an antiviral, an immunostimulatory agent, a corticosteroid, a non-steroidal anti-inflammatory drug (NSAID), a bronchodilator or a mucus viscosity reducing agent, or may be a further excipient. Representative examples of these active agents and excipients are discussed herein.
  • dry it is meant substantially, e.g. essentially, water-free (moisture-free). This may be expressed as a water content of less than 15% w/w, e.g. less than 12%, 10%, 9%, 8%, 7%, 6%, 5%, 4.5%, 4%, 3.5%, 3%, 2.5%, 2%, 1.5% or 1% w/w as measured by weight loss on drying or chemically by the Karl Fischer method (United States Pharmacopeia; European Pharmacopoeia).
  • the particles of the invention, or compositions comprising the particles of the invention may be provided in capsules or other containers, e.g. for use in inhalers.
  • Representative capsule forming substances include but are not limited to methyl acrylate-methacrylic acid copolymers, methyl methacrylate-methacrylic acid copolymers, cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), cellulose acetate trimellitate, and sodium alginate polymer.
  • PVAP polyvinyl acetate phthalate
  • the invention further provides a dry powder inhaler (i.e. an inhaler device) comprising the particles of the invention, dry powder compositions comprising the particles of the invention and/or capsules comprising the same.
  • a dry powder inhaler i.e. an inhaler device
  • the dry powder inhaler may have a reservoir or chamber container containing the particles of the invention or dry powder compositions comprising the particles of the invention
  • the inventors have also developed a surprisingly efficient and cost-effective spray drying process that results in high yields of the advantageous particles of the invention and which, without wishing to be bound by theory, is believed to be made possible by exploiting the particular combination of physical and chemical properties of the alginate oligomer and the selected phospholipid excipient in the particles of the invention.
  • the anti-adherent compound may further contribute to or facilitate this combination of properties.
  • the process exploits an organic-in-aqueous liquid emulsion to maximise the availability of the (limitedly water-soluble) alginate oligomer in the mixture to be spray dried and thereby reach the high levels of alginate oligomer in the particles of the invention for minimum cost of materials (in particular, the organic liquid). It is not typical to approach the use of such a co-solvent system in this way in the context of preparing spray dried particles comprising components of limited water solubility. In such contexts it is more usual to prepare an aqueous-in-organic emulsion in order to increase the concentration of the poorly water soluble components in the mixture to be spray dried.
  • this atypical approach of the invention is made viable on account of the electrostatic interactions that occur between the negatively charged alginate oligomer and the positively charged head groups on the phospholipid stabilising the atypical organic-in-aqueous emulsion.
  • the anti-adherent compound may further contribute to this stabilisation effect.
  • the invention provides a method for the preparation of an organic-in-aqueous liquid emulsion for spray drying so as to form the spray dried particles as defined herein, said method comprising
  • step (c) contains amounts of alginate oligomer, phospholipid and anti-adherent compound and, optionally, further excipients which upon spray drying (which process removes the aqueous and organic liquids but not the various components comprised therein) will form the spray dried particles of the invention, i.e. spray dried particles for inhalation consisting of
  • the aqueous liquid composition(s) and the organic liquid composition may be combined in a single step or a plurality of steps.
  • the combination of the organic liquid composition and the aqueous liquid compositions may be achieved in any order.
  • the aqueous liquid composition(s) may be added to the organic liquid composition
  • the organic liquid composition may be added to the aqueous liquid composition(s) or each element may be combined simultaneously.
  • the organic liquid composition is added to one or more of the aqueous liquid composition(s).
  • an organic-in-aqueous liquid emulsion for spray drying obtained or obtainable from the above method.
  • the organic-in-aqueous liquid emulsion is preferably a substantially stable emulsion, by which it is meant that the emulsion will persist for at least 10 min, preferably at least 15 min, 30 min, 60 min, 120 min or 240 min following cessation of homogenisation.
  • the invention provides a method for the preparation of the spray dried particles as defined herein, said method comprising performing the method for the preparation of an organic-in-aqueous liquid emulsion for spray drying as defined above and further
  • the aqueous liquid composition comprising said alginate oligomer and/or said anti-adherent compound may be formed upon the dissolution of the alginate oligomer and/or anti-adherent compound in any pharmaceutically acceptable aqueous liquid.
  • this liquid will be water, preferably substantially pure water, e.g. distilled water or filtered water, however, some additional compounds (e.g. salts, organic acids, buffers) may also be present so long as such compounds are present in amounts that will not result in spray dried particles containing more than 10% of said compounds and do not interfere with particle formation.
  • the method of the invention may include further steps of preparing the aqueous liquid composition(s). These steps may involve dissolving the alginate oligomer and/or the anti-adherent compound in the aqueous liquid at the same time or sequentially, or adding one of those two ingredients to an aqueous liquid already containing the other.
  • the organic liquid composition comprising said phospholipid may be formed upon the dissolution of the phospholipid in any pharmaceutically acceptable organic liquid, or organic solvent, e.g. alcohols and their derivatives (e.g. methanol, ethanol, C 3 alcohols and C 4 alcohols), ketones (e.g. acetone), acetates (e.g. ethyl acetate), halogenated solvents and their derivatives (e.g. dichloromethane, chloroform), aliphatic solvents and their derivatives (e.g. heptane, hexane, pentane). Ethanol is preferred.
  • the selected organic liquid should have a boiling point at standard atmospheric pressure (1 atm; approximately 101325 Pa) of about 40-100° C.
  • Substantially water miscible organic liquids may be preferable. However, some additional compounds (e.g. salts, organic acids, buffers) may also be present so long as such compounds are present in amounts that will not result in a spray dried particles containing more than 10% of said compounds and do not interfere with particle formation.
  • the method of the invention may include further steps of preparing the organic liquid composition. These steps may involve dissolving the phospholipid in the organic liquid composition.
  • the further excipients of the spray dried particles of the invention may be included in the aqueous liquid composition(s) or the organic liquid composition depending on the hydrophobicity of the further excipient or provided in separate liquid compositions prior to the step of homogenisation.
  • Homogenisation can be achieved by any convenient means, e.g. by mechanical means and/or ultrasonic means, for example by high shear mixing.
  • Apparatus for performing such a homogenisation e.g. a homogeniser
  • a homogeniser is widely known and available in the art, for example a Silverson homogeniser, which may be operated at 10,000 rpm.
  • Homogenisation is preferably performed for such time as a complete emulsion is formed to the naked eye.
  • Homogenisation may be performed as the two liquid phases are combined, or after all of the liquid phases have been combined. In other embodiments homogenisation may begin once a part of each phase has been combined.
  • the volume of the organic liquid composition to be combined with, preferably added to, the aqueous liquid composition(s) must be smaller than that of the total volume of aqueous liquid composition(s).
  • the volume of the organic liquid composition will be less than 95% that of the total volume of aqueous liquid composition(s), e.g. less than 90, 85, 80, 75, 70, 65, 60, 55, 50, 45, 40, 35, 30, 25, 20, 15, 10 or 5% that of the total volume of aqueous liquid composition(s).
  • the volume of the organic liquid composition will be 5-70, 10-65, 15-60, 20-55, 25-50, 30-45, 35-40, 5-60, 10-55, 15-50, 20-45, 25-40, 30-35, 5-50, 10-45, 15-40, 20-35, 25-30, 5-40, 10-35, 15-30, 20-25% that of the total volume of aqueous liquid composition(s). More preferably the volume of the organic liquid composition will be about a third, e.g. 30-35% that of the total volume of aqueous liquid composition(s). In other embodiments the volume of organic liquid composition will be about 15-40%, e.g.
  • the ratio of organic liquid composition aqueous liquid composition(s) is 20-40:60-80, e.g. 20-35:65-80, 20-30:70-80 or 25:75.
  • the method of the invention advantageously allows a reduced amount of organic liquid composition (or organic solvent to be used). The solubility of the alginate oligomer and the desired proportion of alginate oligomer in the final particles may influence the proportions of liquid compositions used.
  • solubility of the other ingredients and the desired proportion of those ingredients in the final particles may also influence the volume of the organic liquid composition which will be used.
  • the proportions of liquid compositions used must be appropriate to result in the formation of a liquid emulsion in the presence of the particle components upon homogenisation.
  • Spray drying can be performed using any convenient technique.
  • the skilled man would be able to adjust his equipment and its use, e.g. the feed rate (and e.g. the inlet temperature, fan speed and pump speed, as appropriate), to ensure the particles of the invention are formed.
  • the methods of the invention may include further formulation and/or biological contaminant removal steps, e.g. combination with further excipients (e.g. powders) or filling into capsules and/or pasteurisation or radiation treatment.
  • the powders of the invention may be radiolabelled, e.g. with technetium-99m, e.g. with Technegas.
  • the particles of the invention may be provided for use in therapy, in particular in the treatment or prevention of respiratory infections and respiratory disorders, especially disorders or conditions which involve microbial infection, in particular biofilm infection, and/or abnormal mucus, e.g. any condition associated with or characterised by defective CFTR ion channel function, COPD, COAD, COAP, bronchitis, cystic fibrosis, a medical disorder or condition associated with CF, emphysema, lung cancer, asthma or pneumonia, or a complication thereof.
  • microbial infection in particular biofilm infection
  • abnormal mucus e.g. any condition associated with or characterised by defective CFTR ion channel function, COPD, COAD, COAP, bronchitis, cystic fibrosis, a medical disorder or condition associated with CF, emphysema, lung cancer, asthma or pneumonia, or a complication thereof.
  • a condition associated with or characterised by defective CFTR ion channel function includes conditions arising from said defective CFTR ion channel function or complications thereof.
  • a “defective CFTR ion channel” will be understood from the above to include any defect or deficiency in CFTR function, i.e. CFTR dysfunction.
  • a defective CFTR ion channel effectively means, and may alternatively be expressed as, “defective CFTR ion channel function”.
  • the condition may thus be viewed as a condition associated with or characterised by or arising from CFTR dysfunction. This may include CFTR ion channels which are defective in the sense that they are non-functional or have reduced function, i.e. partially or fully lack CFTR ion channel activity (in other words in which CFTR ion channel activity is reduced or abrogated).
  • Defective CFTR function may arise from genetic defect or mutation or may be acquired in any other way.
  • CF cystic fibrosis
  • CFTR dysfunction is seen in subjects that have non-compound heterozygous mutant CFTR alleles. In such subjects the inherited dysfunction is mild and so is insufficient to manifest as overt CF, but is sufficient to result the plethora of clinical conditions and complications associated with overt CF.
  • Acquired CFTR dysfunction may arise due to environmental and/or clinical exposure, e.g. through the chronic inhalation of particulate irritants, e.g. smoke particles (tobacco, wood etc.), pollution, dust (asbestos, cotton, coal, stone, animal droppings etc.) and spores.
  • particulate irritants e.g. smoke particles (tobacco, wood etc.), pollution, dust (asbestos, cotton, coal, stone, animal droppings etc.) and spores.
  • a condition associated with or characterised by a defective CFTR ion channel may include not only CF, but also other conditions involving respiratory dysfunction (more generally other respiratory disorders), and in particular disorders involving pulmonary obstruction, including particularly asthma, or respiratory disorders characterised by a chronic inflammatory state, airway remodelling and exacerbations due to respiratory tract infections.
  • Such conditions include non-compound CFTR gene mutation heterozygosity, abnormal mucus clearance in the respiratory tract and/or breathing difficulties resulting from chronic particulate inhalation, COPD, chronic bronchitis, emphysema, bronchiectasis, asthma or chronic sinusitis, or a complication thereof.
  • condition may be a complication, in particular a mucus-related complication, of the above-listed conditions.
  • the invention provides a treatment for mucus stasis and breathing difficulties in tobacco smokers and other subjects exposed to the chronic inhalation of particulate irritants, e.g. smoke particles (tobacco, wood etc.), pollution, dust (asbestos, cotton, coal, stone, animal droppings etc.) and spores.
  • particulate irritants e.g. smoke particles (tobacco, wood etc.), pollution, dust (asbestos, cotton, coal, stone, animal droppings etc.) and spores.
  • a method for the treatment or prevention of a respiratory infection or a respiratory disorder comprising administering the particles of the invention to the airways, preferably the lungs, of a subject in need thereof by inhalation.
  • the invention provides a spray dried particle as defined herein for use in the treatment or prevention of a respiratory infection or a respiratory disorder, said treatment or prevention comprising administering the particles of the invention to the airways, preferably the lungs, of a subject in need thereof by inhalation.
  • the invention provides the use of a spray dried particle as defined herein in the manufacture of a medicament for use in the treatment or prevention of a respiratory infection or a respiratory disorder, said treatment or prevention comprising administering the particles of the invention to the airways, preferably the lungs, of a subject in need thereof by inhalation.
  • the particles of the invention may be used in these aspects together with further pharmaceutical agents (i.e. therapeutic or active agents) or treatment techniques.
  • the particle of the invention maybe used together with an antibiotic, an antifungal, an antiviral, an immunostimulatory agent, a corticosteroid, a non-steroidal anti-inflammatory drug (NSAID), a bronchodilator, a mucus viscosity reducing agent (i.e. an agent which reduces the viscosity of mucus and which terms are used interchangeably with the term “mucolytic”) or a CFTR modulator (also known as a “CFTR modifier”).
  • antibiotic an antifungal, an antiviral, an immunostimulatory agent, a corticosteroid, a non-steroidal anti-inflammatory drug (NSAID), a bronchodilator, a mucus viscosity reducing agent (i.e. an agent which reduces the viscosity of mucus and which terms are used interchangeably with the term “muco
  • the antibiotic may be selected from the aminoglycosides (e.g. amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin); the ⁇ -lactams (e.g. the carbecephems (e.g. loracarbef); the 1st generation cephalosporins (e.g. cefadroxil, cefazolin, cephalexin); 2nd generation cephalosporins (e.g. cefaclor, cefamandole, cephalexin, cefoxitin, cefprozil, cefuroxime); 3rd generation cephalosporins (e.g.
  • aminoglycosides e.g. amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin
  • the ⁇ -lactams e.g. the carbecephems (
  • mafenide sulfacetamide, sulfamethizole, sulfasalazine, sulfisoxazole, trimethoprim-sulfamethoxazole); the tetracyclines (e.g. demeclocycline, doxycycline, minocycline, oxytetracycline, tetracycline); the glycylcyclines (e.g. tigecycline); the carbapenems (e.g.
  • antibiotics include chloramphenicol; clindamycin, ethambutol; fosfomycin; isoniazid; linezolid; metronidazole; nitrofurantoin; pyrazinamide; quinupristin/dalfopristin; rifampin; spectinomycin; and vancomycin.
  • the antibiotic is selected from amikacin, gentamicin, kanamycin, neomycin, netilmicin, streptomycin, tobramycin, cefixime, cefdinir, cefditoren, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepime, aztreonam, amoxicillin, ampicillin, carbenicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, ticarcillin, ciprofloxacin, enoxacin, gatifloxacin, levofloxacin, lomefloxacin, moxifloxacin, norfloxacin, ofloxacin, trovafloxacin, azithromycin, clarithromycin, dirithromycin,
  • the antibiotic is selected from aztreonam, ciprofloxacin, gentamicin, tobramycin, amoxicillin, colistin, ceftazidime, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, spiramycin, oxytetracycline, and imipenem.
  • the antibiotic is selected from aztreonam, ciprofloxacin, gentamicin, tobramycin, amoxicillin, colistin and ceftazidime.
  • Representative antifungals include, but are not limited to the polyenes (e.g. natamycin, rimocidin, filipin, nystatin, amphotericin B, candicin; the imidazoles (e.g. miconazole, ketoconazole, clotrimazole, econazole, bifonazole, butoconazole, fenticonazole, isoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole); the triazoles (e.g.
  • allylamines e.g. terbinafine, amorolfine, naftifine, butenafine
  • echinocandins e.g. anidulafungin, caspofungin, micafungin.
  • antivirals include, but are not limited to abacavir, acyclovir, adefovir, amantadine, amprenavir, arbidol, atazanavir, atripla, boceprevir, cidofovir, combivir, darunavir, delavirdine, didanosine, docosanol, edoxudine, efavirenz, emtricitabine, enfuvirtide, entecavir, famciclovir, fomivirsen, fosamprenavir, foscarnet, fosfonet, ganciclovir, ibacitabine, imunovir, idoxuridine, imiquimod, indinavir, inosine, interferon type III, interferon type, II interferon type I, lamivudine, lopinavir, loviride, maraviroc, moroxydine, nelfinavir, nevirapine
  • immunostimulatory agents include, but are not limited to cytokines e.g. TNF, IL-1, IL-6, IL-8 and immunostimulatory alginates, such as high M-content alginates as described for example in U.S. Pat. No. 5,169,840, WO91/11205 and WO03/045402 which are explicitly incorporated by reference herein in their entirety, but including any alginate with immunostimulatory properties.
  • cytokines e.g. TNF, IL-1, IL-6, IL-8
  • immunostimulatory alginates such as high M-content alginates as described for example in U.S. Pat. No. 5,169,840, WO91/11205 and WO03/045402 which are explicitly incorporated by reference herein in their entirety, but including any alginate with immunostimulatory properties.
  • NSAIDs include, but are not limited to, the salicylates (e.g. aspirin (acetylsalicylic acid), choline magnesium trisalicylate, diflunisal, salsalate, the propionic acid derivatives (e.g. ibuprofen, dexibuprofen, dexketoprofen, fenoprofen, flurbiprofen, ketoprofen, loxoprofen naproxen, oxaprozin), the acetic acid derivatives (e.g.
  • aceclofenac diclofenac, etodolac., indomethacin, ketorolac, nabumetone, tolmetin, sulindac
  • the enolic acid derivatives e.g. droxicam, isoxicam, lornoxicam, meloxicam, piroxicam, tenoxicam
  • the anthranilic acid derivatives e.g. flufenamic acid, meclofenamic acid, mefenamic acid, tolfenamic acid
  • COX-2 inhibitors e.g.
  • the propionic acid derivatives e.g. ibuprofen, dexibuprofen, dexketoprofen, fenoprofen, flurbiprofen, ketoprofen, loxoprofen naproxen, oxaprozin
  • ibuprofen being most preferred.
  • mucus viscosity reducing agent are intended to encompass agents which reduce the intrinsic viscosity of mucus and agents which reduce the attachment of mucus to underlying epithelium, in particular agents which directly or indirectly disrupt the molecular interactions within or between the components of mucus, agents which affect the hydration of mucus and agents which modulate the ionic microenvironment of the mucosal epithelium (particularly the levels of divalent cations, e.g. calcium).
  • suitable mucus viscosity reducing agents include but are not limited to a nucleic acid cleaving enzyme (e.g.
  • a DNase such as DNase I or dornase alfa
  • hypertonic saline gelsolin
  • a thiol reducing agent an acetylcysteine
  • an uncharged low molecular weight polysaccharide e.g. dextran, mannitol
  • arginine or other nitric oxide precursors or synthesis stimulators
  • an agonist of the P2Y2 subtype of purinergic receptors e.g. denufosol
  • an anionic polyamino acid e.g. poly ASP or poly GLU
  • Ambroxol, romhexine, carbocisteine, domiodol, eprazinone, erdosteine, letosteine, mesna, neltenexine, sobrerol, stepronin, tiopronin are specific mucolytics of note. DNase I and hypertonic saline are preferred.
  • Suitable bronchodilators include but are not limited to the ⁇ 2 agonists (e.g. the short-acting ⁇ 2 agonists (e.g. pirbuterol, epinephrine, salbutamol, levosalbutamol, clenbuterol, terbutaline, procaterol, metaproterenol, fenoterol, bitolterol mesylate, ritodrine, isoprenaline); the long-acting ⁇ 2 agonists (e.g. salmeterol, formoterol, bambuterol, clenbuterol); and the ultra-long-acting ⁇ 2 agonists (e.g. indacaterol)), the anticholinergics (e.g. ipratropium, oxitropium, tiotropium) and theophylline.
  • the short-acting ⁇ 2 agonists e.g. pirbuterol, epinep
  • corticosteroids include but are not limited to prednisone, flunisolide, triamcinolone, fluticasone, budesonide, mometasone, beclomethasone, amcinonide, budesonide, desonide, fluocinonide, fluocinolone, halcinonide, hydrocortisone, cortisone, tixocortol, prednisolone, methylprednisolone, prednisone, betamethasone, dexamethasone, fluocortolone, aclometasone, prednicarbate, clobetasone, clobetasol, and fluprednidene.
  • CFTR modulators are small molecules which can redress, at least partially, a CFTR dysfunction.
  • Present CFTR modulators fall into three main groups: CFTR potentiators, CFTR correctors and read-through agents (Derichs, N., Eur. Respir. Rev., 2013, 22(127), 58-65; Petit, R. S. and Fellner, C., Pharmacy and Therapeutics, 2014, 39(7), 500-511; the contents of which are incorporated herein by reference).
  • CFTR potentiators are CFTR modulators which increase the activity of the CFTR ion channel present on the epithelial cell surface.
  • Prototypical examples of CFTR potentiators are ivacaftor (VX-770; N-(2,4-di-tert-butyl-5-hydroxyphenyl)-1,4-dihydro-4-oxoquinoline-3-carboxamide) and VRT-532 (4-methyl-2-(5-phenyl-1H-pyrazol-3-yl)-phenol) of Vertex PharmaceuticalsTM).
  • CFTR correctors are CFTR modulators which increase the amount of CFTR protein delivered or retained at the epithelial cell surface.
  • Prototypical examples of CFTR correctors include lumacaftor (VX-809) and VX-661 of Vertex PharmaceuticalsTM and N6022 (3-[1-(4-carbamoyl-2-methylphenyl)-5-(4-imidazol-1-ylphenyl)pyrrol-2-yl]propanoic acid).
  • Read-through agents also known as “premature stop codon suppressors” (PSC suppressors) or “premature termination codon suppressors” (PTC suppressors, which terms are used interchangeably herein) are CFTR modulators which cause the translation machinery of the cell to pass over any premature termination codon in the CFTR mRNA thereby increasing the amount of substantially full length and functional CFTR produced.
  • PSC suppressors premature stop codon suppressors
  • PTC suppressors premature termination codon suppressors
  • Prototypical examples of read-through agents include ataluren (PTC124) of PTC Therapeutics and gentamicin.
  • CFTR modulators are disclosed in WO2006002421, WO2007056341, WO2007134279, WO2009038683, WO2009064959, WO2009073757, WO2009076141, WO2009076142, WO2010019239, WO2010037066, WO2010048526, WO2010053471, WO2010054138, WO2010138484, WO2011019413, WO2011050325, WO2011072241, WO2011127241, WO2011127290, WO2011133751, WO2011133951, WO2011133953, WO2011133956, WO2011146901, Pedemonte, N., et al., J Clin Invest.
  • the further pharmaceutical agent may conveniently be applied before, simultaneously with or following the particles of the invention. Conveniently the further pharmaceutical agent is applied at substantially the same time as the particles of the invention or afterwards. In other embodiments the further pharmaceutical agent may conveniently be applied or administered before the particles of the invention.
  • the further pharmaceutical agent can also be given (e.g. administered or delivered) repeatedly at time points appropriate for the agent used. The skilled person is able to devise a suitable dosage regimen. In long term treatments the particles of the invention can also be used repeatedly.
  • the particles of the invention can be applied as frequently as the further pharmaceutical agent, or more or less frequently.
  • the particles of the invention and the further pharmaceutical agent may for example be administered together, in a single pharmaceutical formulation or composition, or separately (i.e. separate, sequential or simultaneous administration).
  • the particles of the invention and the further pharmaceutical agent may be combined, e.g. in a pharmaceutical kit or as a combined (“combination”) product.
  • the invention therefore also provides products (e.g. a pharmaceutical kit or a combined (“combination”) product) or compositions (e.g. a pharmaceutical composition) wherein the product or composition comprises the particles of the invention as herein defined and a further pharmaceutical agent, e.g. those described above.
  • a pharmaceutical kit or a combined (“combination”) product e.g. a pharmaceutical kit or a combined (“combination”) product
  • compositions e.g. a pharmaceutical composition
  • the product or composition comprises the particles of the invention as herein defined and a further pharmaceutical agent, e.g. those described above.
  • a further pharmaceutical agent e.g. those described above.
  • Combinations comprising the particles of the invention and an antibiotic, an antifungal, an NSAID, a bronchodilator, a corticosteroid and/or a mucus viscosity reducing agent are preferred.
  • excipients examples include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, inert alginate polymers, tragacanth, gelatine, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, water, water/ethanol, water/glycol, water/polyethylene, hypertonic salt water, glycol, propylene glycol, methyl cellulose, methylhydroxybenzoates, propyl hydroxybenzoates, talc, magnesium stearate, mineral oil or fatty substances such as hard fat or suitable mixtures thereof. Additionally lubricating agents, wetting agents, emulsifying agents, suspending agents, preserving agents, sweetening agents, flavouring agents, and the like may be used.
  • the subject may be any human or animal subject, but more particularly may be a human or a non-human vertebrate, e.g. a non-human mammal, bird, amphibian fish or reptile.
  • the animal may be a livestock or a domestic animal or an animal of commercial value, including laboratory animals or an animal in a zoo or game park. Representative animals therefore include dogs, cats, rabbits, mice, guinea pigs, hamsters, horses, pigs, sheep, goats and cows. Veterinary uses of the invention are thus covered.
  • the subject may be viewed as a patient.
  • the subject is a human.
  • Treatment when used in relation to the treatment of a medical condition/infection in a subject in accordance with the invention is used broadly herein to include any therapeutic effect, i.e. any beneficial effect on the condition or disorder or in relation to the infection/condition/disorder.
  • any therapeutic effect i.e. any beneficial effect on the condition or disorder or in relation to the infection/condition/disorder.
  • eradication or elimination of the infection, or cure of the subject or infection but also an improvement in the infection or condition or disorder of the subject.
  • Treatment thus includes both curative and palliative therapy, e.g. of a pre-existing or diagnosed infection/condition/disorder, i.e. a reactionary treatment.
  • Prevention refers to any prophylactic or preventative effect. It thus includes delaying, limiting, reducing or preventing the condition or disorder (which reference includes infection) or the onset of the condition or disorder, or one or more symptoms or indications thereof, for example relative to the condition or disorder or symptom or indication prior to the prophylactic treatment. Prophylaxis thus explicitly includes both absolute prevention of occurrence or development of the condition or disorder, or symptom or indication thereof, and any delay in the onset or development of the condition or disorder or symptom or indication, or reduction or limitation on the development or progression of the condition or disorder or symptom or indication.
  • Treatment when used in the particular context of the treatment of a CF or a medical disorder or condition associated with CF in accordance with the invention is used broadly herein to include any therapeutic effect, i.e. any beneficial effect on CF or an associated medical disorder or condition or symptom or indicator thereof.
  • a reference to a CF-associated disorder or condition is interchangeable with a reference to a complication of CF.
  • CF is a genetic disease which is characterised in each patient by the unique collection of CF-associated disorders and conditions displayed by the patient at the time of receiving the treatments of the invention
  • treatment of CF can be considered to be the treatment of any or all of the disorders and conditions of the patient or the treatment of a subset thereof.
  • the treatments of the invention described herein do not correct the underlying genetic defect of CF, such treatments are intended to address the effects in the body which arise from the defect, e.g. an alleviation of the effects thereof, e.g. effects arising from the abnormal mucus, and includes the treatment of an associated disorder or condition and also an improvement in the clinical effects of the disorder or condition or overall well-being of the subject.
  • a “cure” of CF would amount to complete alleviation of the various CF-associated disorders and conditions displayed by the patient at the time of receiving the treatments of the invention; however the genetic basis for the disease (the CFTR mutation) would still remain.
  • the treatments of the invention described herein do not require such a “cure” and as noted above, include an improvement in any effect which the CF has on the body.
  • an improvement in any symptom or sign of a CF-associated disorder or condition or in any clinically accepted indicator of a CF-associated disorder or condition in the patient (for example, increasing mucociliary clearance in the lungs, increased responsiveness of lung infections to antibiotics, reduced incidence of constipation or improvement in nutrient absorption).
  • a pre-existing CF-associated disorder or condition is not fully eradicated or the onset of a new CF-associated disorder or condition is not completely halted, but the treatments are sufficient to inhibit these processes to such an extent that the target CF-associated disorder or condition is fully resolved, or at least resolved to some extent, preferably to an extent acceptable to the subject.
  • Treatment in these contexts thus includes both curative and palliative therapy, e.g. of a pre-existing or diagnosed CF-associated disorder or condition, i.e. a reactionary treatment.
  • prevention when used in the particular context of the treatment of CF or a medical disorder or condition associated with CF in accordance with the invention, is used broadly herein to include any prophylactic or preventative effect in the CF patient.
  • a reference to a CF-associated disorder or condition is interchangeable with a reference to a complication of CF.
  • “Prevention” thus includes delaying, limiting, reducing or preventing an effect of CF or a CF-associated or condition or disorder, or one or more symptoms or indications thereof, in a CF patient or the onset of CF or a CF-associated disorder or condition, or one or more symptoms or indications thereof, for example relative to the disorder, condition, symptom or indication thereof prior to the prophylactic treatment.
  • prevention in these contexts thus relates to preventing an effect in the body which arises as a result of the underlying genetic defect, or as a result of the abnormal mucus.
  • CF is a genetic disease which is characterised in each patient by the unique collection of CF-associated disorders and conditions displayed by the patient at the time of receiving the treatments of the invention
  • prevention of CF or a CF-associated disorder or condition in a CF patient can be considered to be the prevention of any CF-associated disorder or condition that the patient has yet to acquire or which the patient has acquired previously but has overcome prior to receiving the claimed treatments.
  • Prophylaxis explicitly includes both absolute prevention of occurrence or development of an effect of CF or a CF-associated disorder or condition, or symptom or indication thereof, and any delay in the onset or development of an effect of CF or a CF-associated disorder or condition, or symptom or indication thereof, or reduction or limitation of the development or progression of CF or a CF-associated disorder or condition, or symptom or indication thereof.
  • the preventative treatments can also be considered as treatments that reduce the risk of a CF patient acquiring or developing CF or a CF-associated disorder or condition, or symptom or indication thereof.
  • Effective delivery is taken to mean delivery of a therapeutically effective amount of alginate oligomer to the lungs, i.e. an amount of alginate oligomer that is capable of treating or preventing a respiratory infection or a respiratory disorder in which microbial infection, in particular biofilm infection, and/or abnormal mucus are involved.
  • FIG. 1 shows a diagram of the apparatus used to radiolabel spray dried particles of the invention with Technegas.
  • Technegas was generated by a Technegas Generator loaded with 99m pertechnetate (400 Mbq) [ 101 ] and was drawn over a bed of dry particles (300 mg) arranged on filter paper [ 102 ] using a vacuum pump [ 103 ]. Free Technegas was trapped in 6% (w/w) EDTA solution [ 104 ].
  • FIG. 2 shows a graphical representation of the mean distribution of weight of radiolabelled and non-radiolabelled particles as well as radioactive doses of radiolabelled particles arising from the aerodynamic size distribution analysis of Example 7.
  • Dark grey bar (left): weight of radiolabelled particles (DPI) n 4.
  • Medium grey bar (middle): radioactive dose of radiolabelled particles (DPI) n 4.
  • Light grey bar (right): weight of non-radiolabelled particles (DPI) n 4.
  • FIG. 3 shows the deposition of Oligo-G in the lungs of a CF patient following administration a nebulised solution ( 3 A (anterior image) and 3 B (posterior image)) or in the form of the spray dried particles of the invention ( 3 C (anterior image) and 3 D (posterior image))
  • FIG. 4 shows the deposition of Oligo-G in the oropharyngeal region of a CF patient following administration a nebulised solution ( 4 A (lateral left) and 4 B (lateral right)) or in the form of the spray dried particles of the invention ( 4 C (lateral left) and 4 D (lateral right))
  • a formulation sample was prepared using compositions described in Table 1. This formulation was scaled up to contain 94 mg/ml concentration of Oligo-G (2600 Da, %G 90-95) in aqueous phase and 80% w/w in solutes concentration. Oligo-G and glycine were dissolved in water to form the aqueous phase. DPPC was dissolved in ethanol to form the organic phase. The organic phase was slowly added to aqueous phase while homogenizing at 10,000 rpm using Silverson homogenizer. After addition of all materials, the suspension formed was homogenized for further 15 minutes.
  • a pilot scale GMP spray drier (Anhydro SPX MS35, USA) was used to spray 1.6 kg of the emulsion prepared above.
  • the powders produced were collected into a glass vial and left to stand overnight. The powders were tested for geometric particle size distribution and APSD using methods described below.
  • the spray dried powder was harvested every 45 minutes to avoid clogging of the neck of the cyclone.
  • the yield was 75% and it took 2.5 hrs to complete.
  • Particle size distribution was determined by laser diffractometry using a Malvern Mastersizer MicroPlus particle size analyzer (Malvern Instruments, UK). Ethyl acetate containing 2 g of SPAN 85 per litre was used as a dispersant for this experiment. The samples were prepared by dispersing around 15 mg of the powder into 3 ml of the dispersant. The equipment was turned on and the following conditions were set.
  • the equipment was left for around one hour to warm up and approximately 100 ml of the dispersant was added to the sample dispersion unit. Backgrounds were first measured using the dispersant. The sample was added drop-wise into the sample dispersion unit until a suitable obscuration value was achieved. Particle size measurements were made after 5 minutes. A minimum of three measurements were made for each sample.
  • Aerodynamic particle size accounts for the geometric particle size, shape and density of the particle. Aerodynamic particle size distribution (APSD) is generally recognized as a critical parameter in the in vitro characterization of inhalation products since it is the APSD of an aerosol cloud that defines where the particles in that cloud are deposited following inhalation. It is generally accepted that drug-containing particles with aerodynamic particle size of less than 5 ⁇ m to be therapeutically effective and the amounts of such particles in a powder (or plurality of particles) is commonly known as FPM (fine particle mass), FPD (fine particle dose) or FPF (fine particle fraction). Particles larger than 5 ⁇ m will generally impact in the oropharynx and be swallowed.
  • FPM fine particle mass
  • FPD fine particle dose
  • FPF fine particle fraction
  • APSD of the powder produced in this study was determined using a next generation impactor (NGI) and HPLC.
  • NGI next generation impactor
  • HPLC HPLC
  • FPM is determined by interpolating NGI results.
  • FPM is estimated as FPM ⁇ 4.46 ⁇ m (i.e. particles with aerodynamic particle size of ⁇ 4.46 ⁇ m) which is calculated by adding the amount of drug deposited from stage 3 to the micro orifice collector (MOC) of the NGI.
  • the device used to generate the aerosol cloud has significant influence on aerosol performance of the formulation.
  • DPI system generally requires high dispersion energy to achieve desired powder de-agglomeration.
  • a DPI system containing micronized powder formulation only requires higher dispersion energy compared to binary powder formulations containing coarse carriers.
  • a high resistance device provides high dispersion energies and it was chosen for this experiment and subsequent product manufacture.
  • Rinsing solvent De-ionized water
  • the powder obtained was tested and the results are shown in Tables 7 and 8. The results demonstrate that powder-containing particles with small geometric particle size distribution that was acceptable for inhalation was obtained. This powder was characterized by acceptable FPM.
  • glycine and leucine were considered as potential anti-adherent agents.
  • Suspension samples were prepared using compositions shown in Table 9. Oligo-G and/or leucine/glycine were dissolved in water to form the aqueous phase.
  • DPPC was dissolved in ethanol to form the organic phase.
  • the aqueous phase was slowly added to organic phase while homogenizing at 6,000 rpm using Silverson homogenizer and microfluidised as described below.
  • a laboratory scale microfluidiser model M-110S (Microfluidics Company, Newton, Mass.) connected to a 110 psi pneumatic supply was used in this process.
  • the mixed material was loaded into the Microfluidiser's bulk vessel and passed through the Microfluidiser interaction chamber before being returned to the top of the bulk chamber. After every cycle/pass the equipment and the suspension was allowed to cool down before further processing. The material was processed for 5 cycles to produce microfluidised suspension.
  • the samples were spray dried using an inlet temperature of 120° C., a fan speed of 50 and pump speed of 10.
  • the powders produced were collected into a glass vial and left to stand overnight.
  • the powders were tested for geometric particle size distribution and APSD using methods described above.
  • Suspension samples were prepared using compositions shown in Table 12 and procedure described in Example 2. The samples were spray dried using parameters described in Example 2. The powders produced were collected into a glass vial and left to stand overnight. The powders were tested for geometric particle size distribution and APSD using methods described Example 1.
  • the low amount of additives and sustained hydrophilicity of the powder of the invention enables very rapid and efficient release of the Oligo-G when exposed to liquid. This was demonstrated in an in vitro setup where the drug product prepared in Example 1 was exposed to saline, at variable concentrations and for different time periods. Approximately 60% of the Oligo-G was released after 1 min, and concentrations of 130 mg/ml, which is close to saturation, could be achieved when small amounts of fluid were applied to the dry powder.
  • the milled powder was assessed using laser diffractometry and NGI to determine APSD, as described in Example 1. Results are summarized in Tables 17 and 18. Although the particle size results are respectable (Table 17). The APSD results (Table 18) demonstrate poor aerosol performance with only around 4 mg of Oligo-G deposited as FPM (target being 15 mg).
  • Dichloromethane was selected as the organic solvent for this experiment.
  • the formulations were prepared and spray-dried using the parameters shown in Table 19. Specifically, DPPC was dissolved in organic solvent and Oligo-G was dissolved in de-ionised water. The aqueous phase was added drop-wise into organic phase while homogenising at 6000 rpm using Silverson homogeniser. Spray-drying was performed while mixing the feed material.
  • Example 1 The spray dried particles prepared in Example 1 were radiolabelled using the apparatus shown in FIG. 1 . Briefly, Technegas generated by a Technegas Generator loaded with 99m pertechnetate (400 Mbq) [ 101 ] was drawn over a bed of dry particles (300 mg) arranged on filter paper [ 102 ] using a vacuum pump [ 103 ]. Free Technegas was trapped in 6% (w/w) EDTA solution [ 104 ].
  • the aerodynamic size distribution of radiolabelled and non-radiolabelled particles was determined by attaching a Miat Monodose Inhaler loaded with a capsule to a multi-stage liquid impinger (MSLI; Copley Scientific, UK).
  • FIG. 2 shows the mean % distribution of radiolabelled and non-radiolabelled particles by weight and by radioactive dose (radiolabelled particles only, measured using the dose calibrator). No radiolabel was detected by this technique in Stage 2, presumably the dose was below the detection level. Radioactive counts of this stage determined by gamma camera imaging confirmed that the % distribution was similar to the radiolabelled and non-radiolabelled particles by weight (approximately 6.8%). The distribution profiles of the three parameters measured were similar, indicating that (a) the radiolabelling process did not affect the aerodynamic properties of the particles; and (b) Technegas adhered uniformly to the particles.
  • the primary objective of this study was to determine, using scintigraphic methods, the lung deposition of Oligo-G when administered to cystic fibrosis patients either as a nebulised solution or in a dry powder form of the spray dried particles of the invention.
  • Secondary objectives were to determine the radiolabel distribution pattern of the two formulations in the diseased lung, including calculating the ratio of radiolabel in the central airways compared to the peripheral region (C/P index); and to characterise the extra-pulmonary deposition (i.e. oropharyngeal and stomach) of radiolabel including retention in the nebuliser or dry powder inhaler reservoir and deposition on the exhalation filter.
  • This study was an open label two-way randomised crossover study in 10 cystic fibrosis patients.
  • the subjects received a single dose of Oligo-G CF-5/20 formulated as spray dried particles of the invention (the spray dried particles of Example 1)—96 mg—delivered by three capsules via the Miat Monodose Dry Powder Inhaler, and a single dose of 1.5 mL (90 mg) aerosolised Oligo-G CF-5/20 6% solution delivered via the Sidestream Plus nebuliser, separated by a 2-14 day washout period.
  • Each treatment was radiolabelled with 10 MBq of 99 mTc in total.
  • Technegas was drawn over a bed of DPI using a vacuum pump, allowing the Tc to adhere to the DPI without affecting its aerodynamic properties (Example 7).
  • Sequential anterior and posterior images of the thorax/abdomen and lateral images of the head/neck were acquired. Additionally, images of the device hardware were acquired pre- and post-dose, using a Siemens E-Cam gamma camera with a 53.3 cm field of view and fitted with a low energy high-resolution collimator.

Landscapes

  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Pulmonology (AREA)
  • Molecular Biology (AREA)
  • Otolaryngology (AREA)
  • Biophysics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Anesthesiology (AREA)
  • Biomedical Technology (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Hematology (AREA)
  • Dispersion Chemistry (AREA)
  • Medicinal Preparation (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
US15/507,712 2014-08-29 2015-08-28 Inhalable powder formulations of alginate oligomers Abandoned US20180235879A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GB1415381.1 2014-08-29
GBGB1415381.1A GB201415381D0 (en) 2014-08-29 2014-08-29 Inhalable powder formulations of alginate oligomers
PCT/EP2015/069785 WO2016030524A1 (en) 2014-08-29 2015-08-28 Inhalable powder formulations of alginate oligomers

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2015/069785 A-371-Of-International WO2016030524A1 (en) 2014-08-29 2015-08-28 Inhalable powder formulations of alginate oligomers

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US17/445,577 Division US11992553B2 (en) 2014-08-29 2021-08-20 Inhalable powder formulations of alginate oligomers

Publications (1)

Publication Number Publication Date
US20180235879A1 true US20180235879A1 (en) 2018-08-23

Family

ID=51752384

Family Applications (2)

Application Number Title Priority Date Filing Date
US15/507,712 Abandoned US20180235879A1 (en) 2014-08-29 2015-08-28 Inhalable powder formulations of alginate oligomers
US17/445,577 Active 2036-06-06 US11992553B2 (en) 2014-08-29 2021-08-20 Inhalable powder formulations of alginate oligomers

Family Applications After (1)

Application Number Title Priority Date Filing Date
US17/445,577 Active 2036-06-06 US11992553B2 (en) 2014-08-29 2021-08-20 Inhalable powder formulations of alginate oligomers

Country Status (14)

Country Link
US (2) US20180235879A1 (pt)
EP (1) EP3185853B1 (pt)
JP (1) JP6707076B2 (pt)
KR (1) KR102582982B1 (pt)
CN (2) CN106659689B (pt)
AU (1) AU2015308381B2 (pt)
BR (1) BR112017004048A2 (pt)
CA (1) CA2959101A1 (pt)
DK (1) DK3185853T3 (pt)
ES (1) ES2972433T3 (pt)
FI (1) FI3185853T3 (pt)
GB (1) GB201415381D0 (pt)
RU (1) RU2708397C1 (pt)
WO (1) WO2016030524A1 (pt)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111202723A (zh) * 2020-02-15 2020-05-29 江苏艾立康药业股份有限公司 一种达芦那韦吸入干粉药物组合物及其制备方法
WO2021183371A1 (en) * 2020-03-11 2021-09-16 Purdue Research Foundation Nano-composite microparticles of polymyxin
WO2023003593A1 (en) * 2021-07-20 2023-01-26 Kenox Pharmaceuticals, Inc. Pulmonary biguanide formulations
WO2023028354A1 (en) * 2021-08-26 2023-03-02 Celestial Therapeutics, Inc Method for generating dry powder suitable for inhalation/intranasal/intratracheal administration of antimicrobial phospholipid compositions

Families Citing this family (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK3219705T3 (da) 2005-12-28 2020-04-14 Vertex Pharma Farmaceutiske sammensætninger af den amorfe form af n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinolin-3-carboxamid
US8563573B2 (en) 2007-11-02 2013-10-22 Vertex Pharmaceuticals Incorporated Azaindole derivatives as CFTR modulators
US8802868B2 (en) 2010-03-25 2014-08-12 Vertex Pharmaceuticals Incorporated Solid forms of (R)-1(2,2-difluorobenzo[D][1,3]dioxo1-5-yl)-N-(1-(2,3-dihydroxypropyl-6-fluoro-2-(1-hydroxy-2-methylpropan2-yl)-1H-Indol-5-yl)-Cyclopropanecarboxamide
ES2858351T3 (es) 2010-04-22 2021-09-30 Vertex Pharma Compuesto intermedio para proceso de producción de compuestos de cicloalquilcaraboxamido-indol
RU2744460C2 (ru) 2014-04-15 2021-03-09 Вертекс Фармасьютикалз Инкорпорейтед Фармацевтические композиции для лечения заболеваний, опосредованных муковисцидозным трансмембранным регулятором проводимости
US10568894B2 (en) 2016-06-03 2020-02-25 Otitopic Inc. Dry powder formulations for inhalation
EP3565848A4 (en) 2017-01-03 2020-09-02 The University of North Carolina at Chapel Hill NITROGEN OXIDE RELEASING ALGINATES AS BIODEGRADABLE ANTIBACTERIAL SCAFFOLDINGS AND ASSOCIATED PROCESSES
GB201702161D0 (en) * 2017-02-09 2017-03-29 Algipharma As A companion diagnostic method for use in the treatment of cystic fibrosis with alginate oligomers
EP3600338A4 (en) 2017-03-28 2020-10-28 The University of North Carolina at Chapel Hill NITRIC OXIDE RELEASING POLYAMINOGLYCOSIDES AS BIODEGRADABLE ANTIBACTERIAL SCAFFOLDS AND RELATED PROCESSES
JP7565588B2 (ja) 2018-03-06 2024-10-11 ザ ユニバーシティ オブ ノース カロライナ アット チャペル ヒル 生分解性の抗菌性足場としての一酸化窒素放出性シクロデキストリンおよびそれに関する方法
JP7354132B2 (ja) * 2018-03-19 2023-10-02 アルギファルマ エーエス マイクロ粒子／ナノ粒子が粘液層を通って移動するのを強化するためのアルギネートオリゴマーの使用
EP3902841A4 (en) 2018-12-28 2022-09-28 The University of North Carolina at Chapel Hill NITRIC OXIDE RELEASING ANTIBACTERIAL POLYMERS AND SCAFFOLDS MADE THEREOF AND METHODS RELATED THEREOF
CN113491678B (zh) * 2020-04-07 2023-01-20 知和(山东)大药厂有限公司 一种盐酸阿比朵尔吸入剂及其制备方法
WO2022029604A1 (en) 2020-08-07 2022-02-10 Council For Scientific And Industrial Research Microemulsion drug delivery system for treatment of acute respiratory distress syndrome

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000033817A1 (en) * 1998-12-08 2000-06-15 Phares Pharmaceutical Research N.V. Phospholipid compositions
US7258873B2 (en) * 2002-04-11 2007-08-21 Medimmune Vaccines, Inc. Preservation of bioactive materials by spray drying
CN102302477A (zh) * 2011-06-02 2012-01-04 张振海 一种华蟾酥毒基干粉吸入剂及其制备方法、应用
US20120065174A1 (en) * 2006-06-30 2012-03-15 Barbara Haeberlin Compositions of glycopyrronium salt for inhalation
US20120115803A1 (en) * 2009-06-03 2012-05-10 Algipharma As Treatment of acinetobacter with alginate oligomers and antibiotics
US20120321717A1 (en) * 2003-04-14 2012-12-20 Vectura Ltd. Devices and pharmaceutical compositions for enhancing dosing efficiency

Family Cites Families (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU7221691A (en) 1990-01-23 1991-08-21 Terje Espevik Mannuronic acid containing alginate wound healing composition and method
US5169840A (en) 1991-03-27 1992-12-08 Nobipols Forskningsstiftelse Diequatorially bound β-1, 4 polyuronates and use of same for cytokine stimulation
NO305033B1 (no) 1997-05-09 1999-03-22 Algipharma As Fremgangsmate for fremstilling av uronsyreblokker fra alginat
EP1148905A2 (de) * 1999-02-03 2001-10-31 Max-Delbrück-Centrum Für Molekulare Medizin Druckluftinhalator zur pulmonalen applikation liposomalen pulver-aerosols sowie dafür geeignete pulver-aerosole
US6586008B1 (en) * 1999-08-25 2003-07-01 Advanced Inhalation Research, Inc. Use of simple amino acids to form porous particles during spray drying
US20100074949A1 (en) 2008-08-13 2010-03-25 William Rowe Pharmaceutical composition and administration thereof
NO326145B1 (no) 2001-11-30 2008-10-06 Fmc Biopolymer As Fremgangsmate for a stimulere vektokning hos pattedyr, fugler og krypdyr.
US8354427B2 (en) 2004-06-24 2013-01-15 Vertex Pharmaceutical Incorporated Modulators of ATP-binding cassette transporters
ES2656017T3 (es) 2004-06-24 2018-02-22 Vertex Pharmaceuticals Incorporated Moduladores de transportadores del casete de unión a ATP
GB0515492D0 (en) * 2005-07-28 2005-08-31 Reckitt Benckiser Healthcare Improvements in or relating to compositions,articles and methods
GB2430881B (en) * 2005-10-06 2010-10-13 Ntnu Technology Transfer As Oligoelectrolyte polyols for the treatment of mucosal hyperviscosity
EP1945632B1 (en) 2005-11-08 2013-09-18 Vertex Pharmaceuticals Incorporated Heterocyclic modulators of atp-binding cassette transporters
ES2377840T3 (es) 2006-05-12 2012-04-02 Vertex Pharmaceuticals, Inc. Composiciones de N-[2,4-bis(1,1-dimetiletil)-5-hidroxifenil]-1,4-dihidro-4-oxoquinolina-3-carboxamida
GB0707096D0 (en) 2007-04-12 2007-05-23 Ntnu Technology Transfer As Method
CA2699292A1 (en) 2007-09-14 2009-03-26 Vertex Pharmaceuticals Incorporated Solid forms of n-[2,4-bis(1,1-dimethylethyl)-5-hydroxyphenyl]-1,4-dihydro-4-oxoquinoline-3-carboxamide
EP3012250B1 (en) 2007-11-16 2017-11-08 Vertex Pharmaceuticals Incorporated Isoquinoline modulators of atp-binding cassette transporters
RU2527894C2 (ru) * 2007-11-27 2014-09-10 Альгифарма ИПР АС Использование альгинатных олигомеров в борьбе с биопленками
JP2011506331A (ja) 2007-12-07 2011-03-03 バーテックス ファーマシューティカルズ インコーポレイテッド ３−（６−（１−（２，２−ジフルオロベンゾ［ｄ］［１，３］ジオキソール−５−イル）シクロプロパンカルボキサミド）−３−メチルピリジン−２−イル）安息香酸の処方物
EP3683218B1 (en) 2007-12-07 2024-09-18 Vertex Pharmaceuticals Incorporated Solid forms of 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioxol-5-yl) cyclopropanecarboxamido)-3-methylpyridin-2-yl) benzoic acid
US20100036130A1 (en) 2007-12-07 2010-02-11 Vertex Pharmaceuticals Incorporated Processes for producing cycloalkylcarboxamido-pyridine benzoic acids
NZ612635A (en) 2007-12-07 2015-06-26 Vertex Pharma Processes for producing cycloalkylcarboxamido-pyridine benzoic acids
EP2243490B1 (en) * 2007-12-28 2012-06-13 Shanghai Institute of Pharmaceutical Industry Insulin nasal powder inhalation
ME03019B (me) 2008-08-13 2018-10-20 Vertex Pharma Farmaceutska kompozicija n-[2,4-bis(1,1- dimeтileтil)-5-hidroksifenil]- 1,4-dihidro-4-oksohinolin-3-karboksamida i njeno ordiniranje
JP2012504143A (ja) 2008-09-29 2012-02-16 バーテックス ファーマシューティカルズ インコーポレイテッド ３−（６−（１−（２，２−ジフルオロベンゾ［ｄ］［１，３］ジオキソール−５−イル）シクロプロパンカルボキサミド）−３−メチルピリジン−２−イル）安息香酸の用量単位
BRPI0919930A2 (pt) 2008-10-23 2016-02-16 Vertex Pharma moduladores de regulador de condutância transmembrana de fibrose cística
NZ592694A (en) 2008-11-06 2013-05-31 Vertex Pharma ATP-Binding Cassette (ABC) transporters as modulators of CFTR activity
UA104876C2 (uk) 2008-11-06 2014-03-25 Вертекс Фармасьютікалз Інкорпорейтед Модулятори atф-зв'язувальних касетних транспортерів
CA2777245A1 (en) 2009-10-22 2011-04-28 Vertex Pharmaceuticals Incorporated Compositions for treatment of cystic fibrosis and other chronic diseases
MX2012011655A (es) 2010-04-07 2012-11-23 Vertex Pharma Formas solidas de acido 3-(6-(1-(2,2-difluorobenzo [d][1-3]dioxol-5-il]ciclopropanocarboxamido)-3-metilpiridin-2-il) benzoico.
HRP20211752T1 (hr) 2010-04-07 2022-02-18 Vertex Pharmaceuticals Incorporated Farmaceutski pripravci 3-(6-(1-(2,2-difluorobenzo[d][1,3]dioksol-5-il)ciklopropankarboksamido)-3-metilpiridin-2-il)benzojeve kiseline i njihova primjena
WO2011133956A1 (en) 2010-04-22 2011-10-27 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions and administrations thereof
NZ603043A (en) 2010-04-22 2015-02-27 Vertex Pharma Pharmaceutical compositions comprising cftr modulators and administrations thereof
ES2858351T3 (es) 2010-04-22 2021-09-30 Vertex Pharma Compuesto intermedio para proceso de producción de compuestos de cicloalquilcaraboxamido-indol
AU2011242452A1 (en) 2010-04-22 2012-11-08 Vertex Pharmaceuticals Incorporated Pharmaceutical compositions and administrations thereof
NZ603386A (en) 2010-05-20 2015-02-27 Vertex Pharma Pharmaceutical compositions and administrations thereof
RU2650636C2 (ru) * 2012-05-03 2018-04-16 Янссен Сайенсиз Айрлэнд Юси Составы на основе полиинозиновой-полицитидиловой кислоты (poly(i:с)) для лечения инфекций верхних дыхательных путей
CA2940560A1 (en) 2014-02-28 2015-09-03 Algipharma As Use of alginate oligomers in the treatment of cystic fibrosis and other conditions associated with defective cftr ion channel function

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000033817A1 (en) * 1998-12-08 2000-06-15 Phares Pharmaceutical Research N.V. Phospholipid compositions
US7258873B2 (en) * 2002-04-11 2007-08-21 Medimmune Vaccines, Inc. Preservation of bioactive materials by spray drying
US20120321717A1 (en) * 2003-04-14 2012-12-20 Vectura Ltd. Devices and pharmaceutical compositions for enhancing dosing efficiency
US20120065174A1 (en) * 2006-06-30 2012-03-15 Barbara Haeberlin Compositions of glycopyrronium salt for inhalation
US20120115803A1 (en) * 2009-06-03 2012-05-10 Algipharma As Treatment of acinetobacter with alginate oligomers and antibiotics
CN102302477A (zh) * 2011-06-02 2012-01-04 张振海 一种华蟾酥毒基干粉吸入剂及其制备方法、应用

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111202723A (zh) * 2020-02-15 2020-05-29 江苏艾立康药业股份有限公司 一种达芦那韦吸入干粉药物组合物及其制备方法
WO2021183371A1 (en) * 2020-03-11 2021-09-16 Purdue Research Foundation Nano-composite microparticles of polymyxin
WO2023003593A1 (en) * 2021-07-20 2023-01-26 Kenox Pharmaceuticals, Inc. Pulmonary biguanide formulations
WO2023028354A1 (en) * 2021-08-26 2023-03-02 Celestial Therapeutics, Inc Method for generating dry powder suitable for inhalation/intranasal/intratracheal administration of antimicrobial phospholipid compositions

Also Published As

Publication number Publication date
CN106659689A (zh) 2017-05-10
KR102582982B1 (ko) 2023-09-25
CN113384538A (zh) 2021-09-14
FI3185853T3 (fi) 2024-02-22
AU2015308381A1 (en) 2017-04-06
CA2959101A1 (en) 2016-03-03
RU2708397C1 (ru) 2019-12-06
JP6707076B2 (ja) 2020-06-10
BR112017004048A2 (pt) 2017-12-05
EP3185853A1 (en) 2017-07-05
JP2017530952A (ja) 2017-10-19
AU2015308381B2 (en) 2020-10-22
US20210386666A1 (en) 2021-12-16
WO2016030524A1 (en) 2016-03-03
GB201415381D0 (en) 2014-10-15
US11992553B2 (en) 2024-05-28
EP3185853B1 (en) 2024-01-31
CN106659689B (zh) 2021-07-13
KR20170044670A (ko) 2017-04-25
ES2972433T3 (es) 2024-06-12
DK3185853T3 (da) 2024-02-26

Similar Documents

Publication Publication Date Title
US11992553B2 (en) Inhalable powder formulations of alginate oligomers
AU2016236172B2 (en) Use of alginate oligomers and CFTR modulators in the treatment of conditions associated with CFTR dysfunction
JP5873616B2 (ja) 乾燥粉末、またはそれから得られた溶液もしくは懸濁液の形態にある吸入用医薬組成物およびその製造方法
PT1274406E (pt) Formulações farmacológicas para inaladores de pó seco sob a forma de pellets duros
HU229073B1 (en) Combinations of formoterol and mometasone furoate for asthma
CN1658837A (zh) 干粉吸入组合物
IL298899A (en) A compound for the treatment of corona virus infections
US10517828B2 (en) Composition comprising at least one dry powder obtained by spray drying to increase the stability of the formulation
TW201306847A (zh) 組成物及用途
EP4178579A1 (en) Glucocorticoid-sparing agent
JP7354132B2 (ja) マイクロ粒子／ナノ粒子が粘液層を通って移動するのを強化するためのアルギネートオリゴマーの使用
KR20170102899A (ko) 약학적 조합물

Legal Events

Date Code Title Description
AS Assignment

Owner name: ALGIPHARMA AS, NORWAY

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:NYAMBURA, BILDAD;BAKLE, ANAND;DESSEN, ARNE;SIGNING DATES FROM 20170224 TO 20170227;REEL/FRAME:041585/0950

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: ADVISORY ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: FINAL REJECTION MAILED

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION