[go: up one dir, main page]

WO2015061412A1 - Méthodes et compositions pour favoriser la dilatation des bronchioles - Google Patents

Méthodes et compositions pour favoriser la dilatation des bronchioles Download PDF

Info

Publication number
WO2015061412A1
WO2015061412A1 PCT/US2014/061717 US2014061717W WO2015061412A1 WO 2015061412 A1 WO2015061412 A1 WO 2015061412A1 US 2014061717 W US2014061717 W US 2014061717W WO 2015061412 A1 WO2015061412 A1 WO 2015061412A1
Authority
WO
WIPO (PCT)
Prior art keywords
surfactant
subject
dppc
amino acid
popg
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.)
Ceased
Application number
PCT/US2014/061717
Other languages
English (en)
Inventor
Ronald A. SIMON
Charles G. Cochrane
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.)
Scripps Research Institute
Original Assignee
Scripps Research Institute
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 Scripps Research Institute filed Critical Scripps Research Institute
Publication of WO2015061412A1 publication Critical patent/WO2015061412A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K31/66—Phosphorus compounds
    • A61K31/683—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols
    • A61K31/685—Diesters of a phosphorus acid with two hydroxy compounds, e.g. phosphatidylinositols one of the hydroxy compounds having nitrogen atoms, e.g. phosphatidylserine, lecithin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/395—Alveolar surfactant peptides; Pulmonary surfactant peptides
    • 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
    • A61K9/0078—Sprays or powders for inhalation; Aerolised or nebulised preparations generated by other means than thermal energy for inhalation via a nebulizer such as a jet nebulizer, ultrasonic nebulizer, e.g. in the form of aqueous drug solutions or dispersions
    • 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
    • A61P11/08—Bronchodilators

Definitions

  • the invention relates to pharmaceutical compositions and methods for treating pulmonary conditions.
  • the primary function of the respiratory system is to provide oxygen to, and remove carbon dioxide from, the circulating blood.
  • Gas exchange between blood and air occurs across the thin walls of tiny hollow sacs called alveoli, which lie within the lung at the distal end of the respiratory tree.
  • the bronchioles represent an important part of the conductive airways that deliver air to the alveoli. Proper bronchiole dilatation and alveoli expansion are essential to the normal function of the respiratory system.
  • asthma Abnormal functions of bronchiole (and possibly also alveoli) are associated with and contribute to many respiratory diseases and disorders such as asthma attack and bronchiolitis.
  • asthma is a result of exposure to allergens such as pollens or house-dust to which the patient has an allergic immune response.
  • Viral infections may also initiate asthma and asthma attacks, such as occurs with respiratory syncytial virus or even the common cold.
  • the symptoms include wheezing, coughing, tightness of the chest, headaches, shortness of breath. These signs and symptoms may suddenly escalate to an acute state, called an asthma attack.
  • Several anatomic causes are responsible for an asthma attack, including bronchoconstriction or bronchospasm and inflammation of the bronchioles, called bronchiolitis. Collapse of areas of the pulmonary alveoli may also exist.
  • Airways can also be restricted by increased production of mucus and edema. All of these decrease the oxygenation of the blood by virtue of the decreased flow of air and loss of pulmonary function.
  • the invention provides methods for promoting dilatation of bronchioles and possibly also expansion of atelectatic alveoli in a subject.
  • the methods are employed to treat subjects who have developed or are risk of developing a pulmonary condition with abnormal or dysfunctional bronchiole dilatation (and possibly also dysfunctional alveoli expansion), e.g., acute asthma.
  • the methods of the invention typically entail administering to a subject afflicted with or at risk of developing the noted condition a
  • the employed pharmaceutical composition is an aerosol.
  • the aerosolized composition can be administered to a subject in need of treatment via, e.g., an aerosol device or a nebulizer.
  • Some methods of the invention can additionally include measuring reduction of air-trapping in the lung of the subject during or after the treatment.
  • the phospholipid in the administered composition is dipalmitoyl phosphatidylcholine (DPPC), palmitoyl-oleoyl phosphatidyl glycerol (POPG), a combination of DPPC and POPG, or a combination of DPPC and POPG plus added palmytic acid.
  • the combination of DPPC and POPG can be a mixture of dipalmitoyl phosphatidylcholine (DPPC) and palmitoyl-oleoyl phosphatidyl glycerol (POPG) in a molar ratio of about 4: 1 to about 2: 1.
  • the administered pharmaceutical composition further contains an effective amount of an isolated surfactant polypeptide.
  • the surfactant polypeptide is a polypeptide having between 5-60 amino acid residues and an amino acid sequence of alternating hydrophobic and hydrophilic amino acid residue regions represented by the formula (Z a U b ) c Z d .
  • Z is a hydrophilic amino acid residue
  • U is a hydrophobic amino acid residue
  • a is an integer with an average value of 1-5
  • b is an integer with an average value of 3-20
  • c is an integer of about 1 to about 10
  • d is an integer of about 0 to about 3.
  • the surfactant polypeptide has an amino acid sequence as shown in any one of SEQ ID NOs: 1-1 1.
  • a preferred surfactant polypeptide for the invention is KLLLLKLLLLKLLLLKLLLLK (SEQ ID NO: 1).
  • the administered pharmaceutical composition is a phospholipid-peptide aqueous dispersion of the surfactant polypeptide and the phospholipid, e.g., the KL4-Surfactant.
  • Methods of the present invention can further include a step of examining the subject for improved bronchiole dilatation, and possibly, alveoli expansion.
  • continuous positive airway pressure (CPAP) is maintained in the subject during and after administration of the pharmaceutical composition.
  • CPAP is provided via a CPAP machine that is coupled to the aerosol device or nebulizer.
  • the CPAP machine can be coupled to the nebulizer via a T-coupled junction tube.
  • the invention provides methods for promoting dilatation of bronchioles and expansion of atelectatic alveoli in a subject.
  • the methods involve administering to the subject a pharmaceutical composition that contains an effective amount of an isolated surfactant polypeptide and a phospholipid.
  • subjects for treatment with these methods are afflicted with or at risk of developing a bronchioli and aveoli related pulmonary condition.
  • the methods are directed to treating subjects who have undergone an acute asthma attack or otherwise have acute inflammation of the bronchial tree.
  • continuous positive airway pressure (CPAP) is maintained in the subject during and after administration of the pharmaceutical composition.
  • the phospholipid present in the administered composition is dipalmitoyl phosphatidylcholine (DPPC), palmitoyl-oleoyl phosphatidyl glycerol (POPG), a combination of DPPC and POPG, or a combination of DPPC and POPG plus added palmytic acid.
  • the employed surfactant polypeptide is KLLLLKLLLLKLLLL LLLLK (SEQ ID NO: l).
  • the administered composition is the KL4-Surfactant.
  • the invention relates to compositions and methods for stimulating or enhancing dilatation and reopening of bronchioles and possibly also expansion of atelectatic alveoli in a subject.
  • Abnormal bronchiole blockage and collapse are associated with, and contribute to the development of, various bronchiole and alveoli related pulmonary conditions, e.g., acute asthma and acute infectious bronchiolitis.
  • Subjects amenable to treatment with methods of the invention include those who are already afflicted with or suffered from a bronchiole and alveoli related pulmonary condition, as well as those who are at risk of developing the condition (e.g., asthma attack or bronchiolitis).
  • the subjects are typically administered with a pharmaceutical composition that contains at least one phospholipid compound.
  • the compositions can also include a surfactant polypeptide described herein.
  • the administered composition can optionally further include other therapeutic agents or excipients.
  • bronchiole related pulmonary conditions such as asthma and bronchiolitis typically suffer from a combination of bronchoconstriction, bronchiolitis, air trapping in the lung as a result of the inflammatory processes associated with these conditions.
  • Administration of the pharmaceutical composition of the invention allows or improves movement of air in and out of the lungs of the subjects.
  • a composition containing phospholipids alone e.g., Dipalmitoyl Phosphatidyl choline, DPPC, and palmitoyl-oleoyl phosphatidyl glycerol, POPG
  • a surfactant polypeptide e.g., the KL 4 -Surfactant composition
  • the subjects can also be applied with continuous positive airway pressure (CPAP) to expand the bronchi and bronchioles, allowing movement of air from the trachea into and out of the pulmonary alveoli.
  • CPAP continuous positive airway pressure
  • the KL 4 -Surfactant and/or phospholipids can also expand atelectatic zones of the lungs.
  • amino acid refers to refers to amino acid residues that can be linked together through formation of a covalent bond between an amino group and a carboxyl group.
  • amino acids can make up a polypeptide or protein. Both genetically-encoded and non-genetically-encoded amino acids are contemplated. Genetically-encoded amino acids are commonly in the natural Inform. However, D-amino acids, substituted amino acids (e.g. , amino acids with modified side chain groups) amino acid metabolites and catabolites, amino acids with "retro" backbones, and amino acid mimics or analogs are also contemplated for use in ⁇ and are thus encompassed by— the present invention. In keeping with standard polypeptide nomenclature, J. Biol. Chem., 243:3557-59, 1969, abbreviations for the more common amino acid residues are as shown in the following Table of Correspondence.
  • amino acid residue sequences represented herein by formulae have a left to right orientation in the conventional direction of amino-terminus to carboxy-terminus.
  • amino acid residue is broadly defined to include the amino acids listed in the Table of Correspondence and modified and unusual amino acids, such as those listed in 37 C.F.R. ⁇ 1.822(b)(4), and incorporated herein by reference.
  • amino acid residue is also broadly defined to include non- genetically-encoded amino acids, D-amino acids, substituted amino acids (e.g., amino acids with modified side chain groups), modified amino acids (e.g., amino acid metabolites, catabolites, and amino acids with "designed” side chains), and amino acid mimics or analogs.
  • a dash at the beginning or end of an amino acid residue sequence generally indicates a bond to a radical such as H and OH (hydrogen and hydroxyl) at the amino- and carboxy-termini, respectively, or a further sequence of one or more amino acid residues.
  • a virgule (/) at the right hand end of a residue sequence indicates that the sequence is continued on the next line.
  • asthma encompasses mild asthma to severe asthma attacks. Asthma attack, also called an asthma exacerbation,
  • asthma attack refers to a sudden worsening of asthma symptoms caused by the tightening of muscles around the airways (bronchospasm).
  • bronchospasm During the asthma attack, the lining of the airways also becomes swollen or inflamed, and thicker mucus (more than normal) is produced. All of these factors, bronchospasm, inflammation, and mucus production, cause symptoms of an asthma attack such as difficulty breathing, wheezing, coughing, shortness of breath, and difficulty performing normal daily activities.
  • Other symptoms of an asthma attack may include severe wheezing when breathing both in and out; coughing that won't stop; very rapid breathing; chest pain or pressure; tightened neck and chest muscles called retractions; difficulty talking; feelings of anxiety or panic; pale, sweaty face; and blue lips or fingernails.
  • Some people with asthma may go for extended periods without having an asthma attack or other symptoms, interrupted by periodic worsening of their symptoms, due to exposure to asthma triggers or perhaps from overdoing it as in exercise-induced asthma.
  • Mild asthma attacks are generally more common. Usually, the airways open up within a few minutes to a few hours after treatment. Severe asthma attacks are less common but last longer and require immediate medical help. It is important to recognize and treat even mild symptoms of an asthma attack to help preventing severe episodes and keep asthma under control.
  • An acute, severe asthma attack that doesn't respond to usual use of inhaled bronchodilators and is associated with symptoms of potential respiratory failure is called status asthmaticus. This is life-threatening and requires immediate medical attention. It is important to be aware of these severe asthma attacks and prevent it with early intervention.
  • the symptoms of a severe asthma attack may include: persistent shortness of breath; the inability to speak in full sentences; breathlessness even while lying down; chest that feels closed; bluish tint to the lips; agitation, confusion, or an inability to concentrate; hunched shoulders and strained abdominal and neck muscles; and a need to sit or stand up to breathe more easily. These are signs of an impending respiratory system failure and require immediate medical attention.
  • the bronchioles or bronchioli are the passageways by which the air passes through the nose or mouth to the air sacs of the lungs in which branches no longer contain cartilage or glands in their submucosa. They are branches of the bronchi.
  • the bronchioles terminate by entering the circular sacs called alveoli.
  • Bronchioles are approximately 1 mm or less in diameter and their walls consist of ciliated cuboidal epithelium and a layer of smooth muscle. Bronchioles divide into even smaller terminal bronchioles that are 0.5 mm or less in diameter.
  • Terminal bronchioles in turn divide into respiratory bronchioles which divide into alveolar ducts. Terminal bronchioles mark the end of the conducting division of air flow in the respiratory system while respiratory bronchioles are the beginning of the respiratory division where actual gas exchange takes place.
  • bronchioles While each smaller than 1 mm in diameter, bronchioles encompass approximately six generations of increasingly finer subdivisions of the bronchi. Terminal bronchioles & respiratory bronchioles (the last 2 generations before the alveoli) can be 0.5 - 0.3 mm in diameter respectively. Unlike bronchi which contain cartilage, bronchioles have no cartilage in their wall, but relatively abundant smooth muscle and elastic fibers. Without the cartilage, bronchioles are subject to collapsing in the absence of pulmonary surfactant either due to lack of production of the peptides (e.g., as in the preemies) or
  • bronchioli related pulmonary conditions or "bronchiole related diseases or disorders” refer to diseases or disorders which are mediated by or associated with abnormal or dysfunctional bronchiolar dilation and, possibly, alveoli expansion during the inhalation/exhalation respiratory cycle. Examples of such conditions include asthma attack, bronchiolitis, chronic bronchitis, obliterans, toxic inhalation, viral and bacterial infections (such as respiratory syncytial virus infections and influenza), and chronic obstructive pulmonary disease.
  • bronchioli related pulmonary conditions for treatment with methods of the present invention are diseases or conditions that involve collapsed bronchioles and possibly also atelectatic alveoli, as opposed to conditions caused by or associated with only abnormal or impaired bronchoconstriction.
  • Some embodiments of the invention are directed to promoting bronchiole dilatation and possibly also alveoli expansion in subjects with bronchiolitis.
  • Bronchiolitis is inflammation of the bronchioles, the smallest air passages of the lungs. It often occurs in children less than two years of age. It presents with coughing, wheezing and shortness of breath which can cause some children difficulty feeding. Treatment is typically supportive. Bronchiolitis is common with up to one third of children being affected in their first year of life.
  • the invention is directed to promoting bronchiole dilatation and possibly also alveoli expansion in subjects with acute asthma attack and/or other acute inflammation of the bronchial tree.
  • bronchiole function is also associated with the development of asthma attack.
  • the bronchiole contracts (bronchospasm) while the lining expands and fluids build up, further constricting the airway.
  • Alveoli refer to an anatomical structure that has the form of a hollow cavity which are located in the respiratory zone of the lungs.
  • the pulmonary alveoli are the terminal ends of the respiratory tree, which outcrop from either alveolar sacs or alveolar ducts, which are both sites of gas exchange with the blood as well.
  • Alveoli are particular to mammalian lungs. Different structures are involved in gas exchange in other vertebrates.
  • the alveolar membrane is the gas-exchange surface. Carbon dioxide rich blood is pumped from the rest of the body into the alveolar blood vessels where, through diffusion, it releases its carbon dioxide and absorbs oxygen.
  • the alveoli contain some collagen and elastic fibers. The elastic fibers allow the alveoli to stretch as they are filled with air during inhalation. They then spring back during exhalation. Abnormal alveoli functions are associated with various diseases, include asthma, chronic bronchitis and pneumonia.
  • Atelectasis refers to the collapse or closure of the lung resulting in reduced or absent gas exchange. It may affect part or all of one lung. It is a condition where the alveoli are deflated, as distinct from pulmonary
  • Atelectasis may be an acute or chronic condition.
  • acute atelectasis the lung has recently collapsed and is primarily notable only for airlessness.
  • chronic atelectasis the affected area is often characterized by a complex mixture of airlessness, infection, widening of the bronchi
  • Continuous positive airway pressure refers to the use of continuous positive pressure to maintain a continuous level of positive airway pressure in a spontaneously breathing patient.
  • CPAP uses mild air pressure to keep an airway open.
  • CPAP typically is used for people who have breathing problems, such as sleep apnea.
  • CPAP also may be used to treat preterm infants whose lungs have not fully developed. For example, doctors may use CPAP to treat infants who have respiratory distress syndrome or bronchopulmonary dysplasia.
  • CPAP treatment involves a CPAP machine, which has three main parts: (1 ) a mask or other device that fits over one's nose and/or mouth; (2) a tube that connects the mask to the machine's motor; and (3) a motor that blows air into the tube.
  • CPAP machines A variety of fixed pressure and automatic CPAP machines are available and can be readily obtained from commercial vendors, e.g., CPAP Supply USA (Chesterfield, VA, USA) and Americare Respiratory Services, Inc. (Santa Ana, CA, USA).
  • human means compatible with human physiology and a noun associated with the term “human” need not be strictly derived from Homo sapiens.
  • a polypeptide or other material that is described as “human” will cause substantially no immune reaction in a human.
  • the surfactant polypeptides of the invention may not all be derived from a human source or may not have an amino acid sequence identical to known human proteins, but such surfactant polypeptides may be referred to as "human” so long as they cause substantially no immune response in a human.
  • Isolated means that the isolated material has been removed from the environment in which it is naturally synthesized. In some embodiments, an “isolated” material has been removed from the natural environment. Thus, an “isolated” material may be present in a composition or another environment where it would not be naturally found. For example, a surfactant polypeptide described herein may be isolated even though it has been mixed into a composition containing other ingredients or is present in a recombinant organism that was used for recombinant production of the polypeptide.
  • KL 4 -Surfactant refers to an aqueous dispersion containing a specific surfactant polypeptide and a phospholipid component.
  • the polypeptide is KL 4 - Surfactant is KL 4 (SEQ ID NO: l).
  • the phospholipid component consists of phospholipids dipalmitoyl phosphatidylcholine (DPPC) and palmitoyl- oleoylphosphatidyl glycerol (POPG), optionally also a fatty acid, palmitic acid.
  • DPPC/POPG palmitoyl- oleoylphosphatidyl glycerol
  • ratio of the two phospholipids (DPPC/POPG) in KL 4 -Surfactant is about 3: 1.
  • “Pharmaceutically acceptable” is a term that refers to molecular entities and compositions that do not produce an allergic or similar untoward reaction when administered to a human.
  • patient and “subject” are used interchangeably and refer to mammals such as human patients and non-human primates, as well as experimental animals such as rabbits, rats, and mice, and other animals.
  • Animals include all vertebrates, e.g., mammals and non-mammals, such as dogs, cats, sheeps, cows, pigs, rabbits, chickens, and etc.
  • Preferred subjects for practicing the therapeutic methods of present invention are human.
  • Subjects in need of treatment include patients already suffering from a bronchiole and alveoli related pulmonary condition as well as those prone to developing the condition.
  • Phospholipids refers to amphipathic lipids that are composed of a nonpolar hydrophobic tail, a glycerol or sphingosine moiety, and a polar head.
  • the nonpolar hydrophobic tail is usually a saturated or unsaturated fatty acid group.
  • the polar head has a phosphate group that is often attached to a nitrogen- containing base.
  • treating includes (i) preventing a disease or disorder (e.g., asthma attack) from occurring (e.g. prophylaxis); (ii) inhibiting or arresting its development; and (iii) relieving symptoms associated with the condition.
  • treatment includes the administration of a pharmaceutical composition of the invention and/or other therapeutic compositions or agents to prevent or delay the onset of the symptoms, complications, or biochemical indicia of the disease described herein, alleviating or ameliorating the symptoms or arresting or inhibiting further development of the disease, condition, or disorder.
  • Treatment further refers to any indicia of success in the treatment or amelioration or prevention of the disease, condition, or disorder described herein, including any objective or subjective parameter such as abatement; remission; or diminishing of symptoms or making the disease condition more tolerable to the patient.
  • Detailed procedures for the treatment or amelioration of the disease can be based on objective or subjective parameters, including the results of an examination by a physician.
  • a “protein” or “polypeptide” or “peptide” is a biopolymer composed of amino acid or amino acid analog subunits, typically some or all of the 20 common L-amino acids found in biological proteins, linked by peptidyl intersubunit linkages, or other intersubunit linkages that do not substantially alter the surfactant activity of the protein, polypeptide or peptide.
  • a protein has a primary structure represented by its subunit sequence, and may have secondary helical or pleat structures, as well as overall three-dimensional structure.
  • protein commonly refers to a relatively large polypeptide, e.g., containing 30 or more amino acids
  • peptide to or “polypeptide” to smaller polypeptides
  • the terms are also used interchangeably herein. That is, the term “protein” may refer to a larger polypeptide, e.g. , greater than 30 amino acids, but does not necessarily exclude a smaller polypeptide, and the term “polypeptide” may refer to a smaller peptide, e.g., fewer than 30 amino acids, but may also include larger proteins.
  • “Purified” means that a material has been removed from the environment in which it was made.
  • a material may be partially or substantially purified and need not be completely (100%) pure.
  • a surfactant polypeptide described herein may be purified after it has been chemically or recombinantly synthesized by removing some or all of the unreacted chemicals, side products, cellular debris and other components.
  • “Surfactant activity” refers to the ability of any substance, such as an organic molecule, protein, peptide or polypeptide, when combined with lipids, either alone or in combination with other organic molecules, to lower surface tension at an air/water interface. The measurement can be made with a
  • Polypeptide surfactant activity can also be assessed in vitro.
  • the pressure across an air-liquid interface (expressed in negative cm of 3 ⁇ 40 pressure) at minimal ( ⁇ min) bubble radius can be measured at various times using the pulsating bubble technique described by Enhorning, J. Appl. Physiol., 43 : 198-203 (1977). Briefly, the Enhorning Surfactometer
  • compositions employed in the methods of the invention contain one or more phospholipid compounds described herein.
  • the phospholipid suitable for the invention can be any phospholipid available to one of skill in the art, including native and/or synthetic phospholipids.
  • the phospholipid component of the compositions of the invention can include one or more phospholipids such as phosphatidylcholine (PC), phosphatidyl
  • phospholipids preferably at least about 7 carbon atoms in length, typically 12-20 carbons in length, and may be entirely saturated or partially unsaturated. It is known that phospholipids, such as DPPC, are absorbed relatively slowly to the air-cell lining interface when administered alone and, once adsorbed, spread slowly.
  • phospholipids useful in the invention include phosphatidylcholines, such as dipalmitoyl phosphatidylcholine (DPPC), dilauryl phosphatidylcholine (DLPC) C 12:0, dimyristoyl phosphatidylcholine (DMPC) C I 4:0, distearoyl phosphatidylcholine (DSPC), diphytanoyl
  • DPPC dipalmitoyl phosphatidylcholine
  • DLPC dilauryl phosphatidylcholine
  • DMPC dimyristoyl phosphatidylcholine
  • DSPC distearoyl phosphatidylcholine
  • phosphatidylcholine nonadecanoyl phosphatidylcholine, arachidoyl phosphatidylcholine, dioleoyl phosphatidylcholine (DOPC) (CI 8: 1), dipalmitoleoyl phosphatidylcholine (C 16: 1), linoleoyl phosphatidylcholine (C I 8:2)), dipalmitoyl phosphatidylethanolamine,
  • DOPE dioleoylphosphatidylethanolamine
  • DOPG dioleoyl phosphatidylglycerol
  • POPG palmitoyloleoyl phosphatidylglycerol
  • DSPS distearoylphosphatidylserine
  • the following phospholipids can be employed: 1 ,2-diacyl- sn-glycero-3-[phospho-rac-( 1 -glycerol)], 1 ,2-diacyl-sn-glycero-3-[phospho-L- serine], 1,2 diacyl-sn-glycero-3-phosphocholine, l ,2-diacyl-sn-glycero-3- phosphate, l ,2-diacyl-sn-glycero-3-phosphoethanolamine where the diacyl groups may be symmetrical, asymmetrical and contain either saturated or unsaturated fatty acids of various types ranging from 3 to 28 carbons in chain length and with up to 6 unsaturated bonds.
  • DPPC is the principal phospholipid in all mammalian species examined to date. DPPC is synthesized by epithelial cells of the airspaces (the type 2 pneumocyte of the alveoli and an as yet unidentified cell of the airways). DPPC is secreted into a cellular lining layer and spreads out to form a monomolecular film over the alveoli.
  • the DPPC film at the air-cellular lining interface has certain unique properties that explain its normal function: (1) the film, which spreads to cover all surfaces, achieves extremely low surface tension upon compression, e.g., during exhalation, thereby reducing the net force that favors liquid movement into the airspace; (2) as airway or alveolar size falls, surface tension falls proportionately, thereby establishing a pressure equilibration among structures to prevent collapse; (3) because of its amphoteric structure, the film can form loose chemical associations with both hydrophobic and hydrophilic moieties and because of its high compressibility these associations can be broken upon film compression, thereby freeing the moiety from the interface; and (4) these loose chemical associations can be modified by the addition of other compounds found in the surfactant system (PG, for example) that can alter the charge distribution on the film, thereby altering the rate at which the moiety (as mentioned in (3) above) is released from the film.
  • PG surfactant system
  • the phospholipid component is DPPC and the DPPC comprises about 50 to about 90 weight percent of the surfactant composition.
  • DPPC comprises about 50 to 75 weight percent of the surfactant composition with about 15 to about 40 percent of the composition comprising unsaturated phosphatidylcholine, phosphatidylglycerol (PG), triacylglycerols, palmitic acid, spingomyelin or admixtures thereof.
  • the phospholipid component is an admixture of DPPC and POPG in a weight ratio of about between 4: 1 and 2: 1.
  • the phospholipid component is an admixture of DPPC and palmitoyl-oleoyl phosphatidylglycerol (POPG) in a weight ratio of about 3 : 1.
  • POPG palmitoyl-oleoyl phosphatidylglycerol
  • a fatty acid molecule, palmytic acid can also be present in the admixture.
  • the phospholipid(s) can make up 50-95 dry weight percent of the surfactant composition, and preferably between 80-90 percent by dry weight of the composition.
  • DPPC and the above-described lipids and phospholipids can be obtained commercially, or prepared according to published methods that are generally known in the art.
  • the pharmaceutical compositions of the invention can additionally contain other therapeutic agents (e.g., surfactant polypeptides or anti-inflammatory agents), as well as other accessory molecules (e.g., spreading agents) and pharmaceutically acceptable excipients as detailed below.
  • therapeutic agents e.g., surfactant polypeptides or anti-inflammatory agents
  • other accessory molecules e.g., spreading agents
  • pharmaceutically acceptable excipients as detailed below.
  • the various components of the pharmaceutical composition can be mixed in dry, solution, or particle-suspension form, and may be preformulated, prior to addition of the therapeutic agents, or may be formulated together with the therapeutic agents.
  • the active ingredient and other agents are formulated as an aerosolized composition.
  • the surfactant polypeptides employed in the invention are polypeptides proteins, and/or peptides that, either alone or in combination with other organic molecules, can lower surface tension at an air/water interface.
  • Surfactant polypeptides suitable for the invention include polypeptides that have amino acid sequences with alternating charged and uncharged amino acid residue regions.
  • Polypeptide surfactants having amino acid residue sequences with alternating hydrophobic and hydrophilic amino acid residue regions are also employed in the compositions and methods of the present invention.
  • the surfactant polypeptides can have at least about 4, or at least about 8, or at least about 10, amino acid residues.
  • surfactant polypeptides are generally not more than about 60 amino acid residues in length, although longer and even full-length native surfactant proteins are also contemplated.
  • Examples of surfactant polypeptides that can be used in the compositions and methods of the invention are described in U.S. Patent No. 6,013,619, U.S. Patent No. 5,789,381 , U.S. Patent No. 5,407,914, U.S. Patent No. 5,260,273 and U.S. Patent No. 5, 164,369, all of which are incorporated by reference herein.
  • the surfactant polypeptides of the present invention can have alternating groupings of charged and uncharged amino acid residues amino acid residues as represented by the formula [(Charged) a (Uncharged)b] c (Charged)d, wherein “a” has an average value of about 1 to about 5; “b” has an average value of about 3 to about 20; “c” is 1 to 10; and “d” is 0 to 3.
  • Organic surfactant molecules not comprised solely of amino acid residues alone preferably have a similar structure constituted by alternating groupings of charged and uncharged (or
  • amino acids can be placed into different classes depending primarily upon the chemical and physical properties of the amino acid side chain. For example, some amino acids can be charged, hydrophilic or polar amino acids and others can be uncharged, hydrophobic or nonpolar amino acids.
  • Polar amino acids include amino acids having acidic, basic or hydrophilic side chains and nonpolar amino acids include amino acids having aromatic or hydrophobic side chains.
  • Nonpolar amino acids may be further subdivided to include, among others, aliphatic amino acids.
  • the definitions of the classes of amino acids as used herein are as follows:
  • Nonpolar Amino Acid refers to an amino acid having a side chain that is uncharged at physiological pH, that is not polar and that is generally repelled by aqueous solution.
  • Examples of genetically encoded hydrophobic amino acids include alanine, leucine, isoleucine, methionine, phenylalanine, tryptophan, tyrosine and valine.
  • cysteine is a nonpolar amino acid.
  • non-genetically encoded nonpolar amino acids include t-BuA, Cha, norleucine, and/or an a-aminoaliphatic carboxylic acid, such as a-aminobutanoic acid, a-aminopentanoic acid, a-amino-2-methylpropanoic acid, or a- aminohexanoic acid.
  • Aromatic amino acid refers to a nonpolar amino acid having a side chain containing at least one ring having a conjugated ⁇ -electron system
  • aromatic group (aromatic group).
  • the aromatic group may be further substituted with substituent groups such as alkyl, alkenyl, alkynyl, hydroxyl, sulfonyl, nitro and amino groups, as well as others.
  • substituent groups such as alkyl, alkenyl, alkynyl, hydroxyl, sulfonyl, nitro and amino groups, as well as others.
  • genetically encoded aromatic amino acids include phenylalanine, tyrosine and tryptophan.
  • Commonly encountered non-genetically encoded aromatic amino acids include
  • phenylglycine 2-naphthylalanine, ⁇ -2-thienylalanine, 1 ,2,3,4- tetrahydroisoquinoline-3-carboxylic acid, 4-chlorophenylalanine, 2- fluorophenylalanine, 3-fluorophenylalanine and 4-fluorophenylalanine.
  • Aliphatic amino acid refers to a nonpolar, uncharged amino acid having a saturated or unsaturated straight chain, branched or cyclic hydrocarbon side chain.
  • genetically encoded aliphatic amino acids include Ala, Leu, Val and He.
  • non-encoded aliphatic amino acids include Nle.
  • Polar amino acid refers to a hydrophilic amino acid having a side chain that is charged or uncharged at physiological pH and that has a bond in which the pair of electrons shared in common by two atoms is held more closely by one of the atoms.
  • Polar amino acids are generally hydrophilic, meaning that they have an amino acid having a side chain that is attracted by aqueous solution.
  • genetically encoded polar amino acids include asparagine, glutamine, lysine and serine.
  • cysteine is a polar amino acid.
  • non-genetically encoded polar amino acids include citrulline, homocysteine, N-acetyl lysine and methionine sulfoxide.
  • Acidic Amino Acid refers to a hydrophilic amino acid having a side chain p value of less than 7. Acidic amino acids typically have negatively charged side chains at physiological pH due to loss of a hydrogen ion. Examples of genetically encoded acidic amino acids include aspartic acid (aspartate) and glutamic acid (glutamate).
  • Basic Amino Acid refers to a hydrophilic amino acid having a side chain pK value of greater than 7.
  • Basic amino acids typically have positively charged side chains at physiological pH due to association with hydronium ion.
  • genetically encoded basic amino acids include arginine, lysine and histidine.
  • non-genetically encoded basic amino acids include the amino acids ornithine, 2,3-diaminopropionic acid, 2,4-diaminobutyric acid and homoarginine.
  • Ionizable Amino Acid or “Charged Amino Acid” refers to an amino acid that can be charged at a physiological pH.
  • Such ionizable or charges amino acids include acidic and basic amino acids, for example, D-aspartic acid, D- glutamic acid, D-histidine, D-arginine, D-lysine, D-hydroxylysine, D-ornithine, D- 3-hydroxyproline, L-aspartic acid, L-glutamic acid, L-histidine, L-arginine, L- lysine, L-hydroxylysine, L-ornithine or L-3-hydroxyproline.
  • tyrosine has both a nonpolar aromatic ring and a polar hydroxyl group.
  • tyrosine has several characteristics that could be described as nonpolar, aromatic and polar.
  • the nonpolar ring is dominant and so tyrosine is generally considered to be nonpolar.
  • cysteine also has nonpolar character.
  • cysteine can be used to confer hydrophobicity or nonpolarity to a peptide.
  • surfactant polypeptides include a sequence having alternating groupings of amino acid residues as represented by the formula (Z a U b ) c Z d , wherein Z is a charged amino acid and U is an uncharged amino acid; "a” has an average value of about 1 to about 5; “b” has an average value of about 3 to about 20, “c” is 1 to 10; and “d” is 0 to 3.
  • Z is histidine, lysine, arginine, aspartic acid, glutamic acid, 5-hydroxylysine, 4-hydroxyproline, and/or 3-hydroxyproline
  • U is valine, isoleucine, leucine, cysteine, tyrosine, phenylalanine, and/or an a- aminoaliphatic carboxylic acid, such as a-aminobutanoic acid, a-aminopentanoic acid, a-amino-2-methylpropanoic acid, or a-aminohexanoic acid.
  • polypeptides of the present invention have alternating groupings or amino acids residue regions as represented by the formula (B a U b ) c B d , wherein B is an amino acid residue independently selected from the group consisting of histidine, lysine, 5-hydroxylysine,
  • B is an amino acid derived from collagen and is selected from the group consisting of 5 -hydroxy lysine, 4-hydroxyproline, and 3-hydroxyproline; "a” has an average value of about 1 to about 5; “b” has an average value of about 3 to about 20; “c” is 1 to 10; and “d” is 0 to 3.
  • surfactant polypeptides of the present invention include a sequence having alternating groupings of amino acid residues as represented by the formula (B a Jt,) c B d , wherein B is an amino acid residue independently selected from the group consisting of histidine,
  • J is an a-aminoaliphatic carboxylic acid having four to six carbons, inclusive. In other embodiments, J is an a-aminoaliphatic carboxylic acid having six or more carbons, inclusive. In yet other variations, J is preferably selected from the group consisting of a-aminobutanoic acid, a-aminopentanoic acid, a-amino-2- methylpropanoic acid, and a-aminohexanoic acid.
  • Another embodiment contains surfactant polypeptides including a sequence having alternating groupings of amino acid residues as represented by the formula (Z a U b ) c Z d , wherein Z is an amino acid residue independently selected from the group consisting of R, D, E, and K; and U is an amino acid residue independently selected from the group consisting of V, I, L, C, Y and F.
  • U is selected from the group consisting of V, I, L, C and F; or from the group consisting of L and C.
  • the integer "a” has an average value of about 1 to about 5; "b” has an average value of about 3 to about 20; “c” is 1 to 10; and "d” is 0 to 3.
  • Z and U, Z and J, D and U, and B and J are amino acid residues that, at each occurrence, are independently selected.
  • "a” generally has an average value of about 1 to about 5;
  • "b” generally has an average value of about 3 to about 20;
  • "c” is 1 to 10; and
  • "d” is 0 to 3.
  • Z and B are charged amino acid residues. In other embodiments, Z and B are hydrophilic or positively charged amino acid residues. In one variation, Z is selected from the group consisting of R, D, E and K. In another embodiment, Z is preferably selected from the group consisting of R and K. In yet another, B is selected from the group consisting of histidine, 5 -hydroxy lysine, 4-hydroxyproline, and 3-hydroxyproline. In another embodiment, B is a collagen constituent amino acid residue and is selected from the group consisting of 5-hydroxylysine, ( ⁇ -hydroxylysine), 4-hydroxyproline, and 3-hydroxyproline. In another embodiment, B is histidine.
  • U and J are uncharged amino acid residues. In some embodiments, U and J are hydrophobic amino acid residues.
  • U is selected from the group consisting of V, I, L, C, Y, and F. In another embodiment, U is selected from the group consisting of V, I, L, C, and F. In yet another embodiment, U is selected from the group consisting of L and C. In various embodiments, U is L.
  • B is an amino acid selected from the group consisting of histidine, 5-hydroxylysine, 4-hydroxyproline, and
  • B may be selected from the group consisting of collagen-derived amino acids, which includes 5-hydroxylysine,
  • charged and uncharged amino acids are selected from groups of modified amino acids.
  • a charged amino acid is selected from the group consisting of citrulline, homoarginine, or ornithine, to name a few examples.
  • the uncharged amino acid is selected from the group consisting of a-aminobutanoic acid, a-aminopentanoic acid, a-amino-2- methylpropanoic acid, and a-aminohexanoic acid.
  • variables "a”, “b”, “c” and “d” are integers that indicate the number of charged or uncharged residues (or hydrophilic or hydrophobic residues).
  • "a” has an average value of about 1 to about 5, or of about 1 to about 3, or of about 1 to about 2, or of about 1.
  • "b" is an integer with an average value of about 3 to about 20, or about 3 to about 12, or about 3 to about 10, or about 4 to about 8. In one embodiment, "b" is about 4.
  • "c” is an integer with an average value of about 1 to about 10, or about 2 to about 10, or about 3 to about 8, or about 4 to about 8, or about 3 to about 6. In one embodiment, "c" is about 4.
  • "d” is an integer with an average value of about 0 to about 3 or about 1 to about 3. In one embodiment, “d” is about 0 to about 2, or 1 to 2; in another embodiment, “d” is 1.
  • an amino acid residue - e.g., a residue represented by Z or U ⁇ is independently selected, it is meant that at each occurrence, a residue from the specified group is selected. That is, when "a" is 2, for example, each of the hydrophilic residues represented by Z will be independently selected and thus can include, for example, RR, RD, RE, RK, DR, DD, DE, DK, etc.
  • surfactant polypeptide that can be used in the compositions and methods of the invention is SEQ ID NO: 12.
  • each Xa is separately selected from lysine or arginine, and each Xb is separately selected from aspartic acid or glutamic acid.
  • the designation is an abbreviation for the indicated amino acid residue sequence.
  • a composite polypeptide consists essentially of an amino terminal sequence and a carboxy terminal sequence.
  • the amino terminal sequence has an amino acid sequence of a hydrophobic region polypeptide or a hydrophobic peptide of this invention, preferably hydrophobic polypeptide, as defined in the above formula.
  • the carboxy terminal sequence has the amino acid residue sequence of a subject carboxy terminal peptide.
  • SP Surfactant Protein
  • proteins and polypeptides derived from or having characteristics similar to those of natural Surfactant Protein (SP) are useful in the present methods.
  • natural surfactant proteins include SP-A, SP-B, SP-C or SP-D, or fragments thereof. Amino acid sequences related to such natural surfactant proteins are known in the art. See, e.g., NCBI database as accession numbers NP 005402 (gi: 13346504), NP 008857 (gi: 13346506), NP 000533 (gi:
  • the surfactant polypeptide of this invention has amino acid residue sequence that has a composite hydrophobicity of less than zero, preferably less than or equal to - 1 , more preferably less than or equal to -2. Determination of the composite hydrophobicity value for a peptide is known in the art, see, U.S. Patent No. 6,013,619, the disclosure of which is incorporated herein by reference.
  • the surfactant of the present invention comprises a peptide-containing preparation, the 21 -residue peptide being a mimic of human SP-B consisting of repeated units of four hydrophobic leucine (L) residues, bounded by basic polar lysine (K) residues.
  • KL 4 has the following amino acid residue sequence: KLLLL LLLLKLLLLKLLLLK (SEQ ID NO 1).
  • Synthetic polypeptides suitable for use in compositions of the present invention can be synthesized from amino acids by techniques that are known to those skilled in the polypeptide art. An excellent summary of the many techniques available may be found in J.M. Steward and J.D. Young, SOLID PHASE PEPTIDE SYNTHESIS, W.H. Freeman Co., San Francisco, 1969, and J. Meienhofer, HORMONAL PROTEINS AND PEPTIDES, Vol. 2, p. 46, Academic Press (New York), 1983 for solid phase peptide synthesis, and E. Schroder and . Kubke, THE PEPTIDES, Vol. 1 , Academic Press (New York), 1965 for classical solution synthesis.
  • these methods comprise the sequential addition of one or more amino acid residues or suitably protected amino acid residues to a growing peptide chain.
  • amino acid residues or suitably protected amino acid residues Normally, either the amino or carboxyl group of the first amino acid residue is protected by a suitable, selectively removable protecting group.
  • a different, selectively removable protecting group is utilized for amino acids containing a reactive side group (e.g., lysine).
  • Example 1 describes a solid phase synthesis of the surfactant peptide, a protected or derivatized amino acid is attached to an inert solid support through its unprotected carboxyl or amino group.
  • the protecting group of the amino or carboxyl group is then selectively removed and the next amino acid in the sequence having the complementary (amino or carboxyl) group suitably protected is admixed and reacted under conditions suitable for forming the amide linkage with the residue already attached to the solid support.
  • the protecting group of the amino or carboxyl group is then removed from this newly added amino acid residue, and the next amino acid (suitably protected) is then added, and so forth.
  • any remaining terminal and side group protecting groups are removed sequentially or concurrently, to afford the final polypeptide. That polypeptide is then washed by dissolving in a lower aliphatic alcohol, and dried. The dried surfactant polypeptide can be further purified by known techniques, if desired.
  • the surfactant proteins and polypeptides of the present invention may also be produced by recombinant DNA technology.
  • the procedure of deriving protein molecules from the plant or animal hosts are generally known in the art. See, Jobe et al., Am. Rev. Resp. Dis., 136: 1032 (1987); Glasser et al, J. Biol. Chem. , 263 : 10326, (1988).
  • a gene sequence encoding the proteins or polypeptides under the control of a suitable promoter and/or signal peptide is inserted into a plasmid or vector for transfection of a host cells.
  • the expressed proteins/polypeptide may be isolated from the cell culture.
  • polypeptides disclosed herein comprise naturally-occurring amino acids in the "L" form that are joined via peptide linkages
  • molecules including amino acid side chain analogs, non-amide linkages may also display a significant surfactant activity and may possess other advantages, as well.
  • a molecule e.g., for use in a surfactant composition
  • Molecules comprising a series of amino acids linked via a "retro" backbone, i.e., a molecule that has internal amide bonds constructed in the reverse direction of carboxyl terminus to amino terminus, are also more difficult to degrade and may thus be useful in various applications, as described herein.
  • a "retro" backbone i.e., a molecule that has internal amide bonds constructed in the reverse direction of carboxyl terminus to amino terminus
  • other groups besides a C3 ⁇ 4 group may be added to the alpha carbon atom, that is, surfactant molecules of the present invention are not limited to those incorporating a CH 3 at the a carbon alone.
  • any of the side chains and molecules described above may be substituted for the indicated CH 3 group at an a carbon component.
  • analogs and “derivatives” of polypeptides and amino acid residues are intended to encompass metabolites and catabolites of amino acids, as well as molecules that include linkages, backbones, side- chains or side-groups that differ from those ordinarily found in what are termed “naturally-occurring” L-form amino acids.
  • analogs and “derivative” may also conveniently be used interchangeably herein.
  • D-amino acids, molecules that mimic amino acids and amino acids with "designed" side chains are also encompassed by the terms “analogs” and
  • a wide assortment of useful surfactant molecules including amino acids having one or more extended or substituted R or R' groups, is also contemplated by the present invention. Again, one of skill in the art should appreciate from the disclosures that one may make a variety of modifications to individual amino acids, to the linkages, and/or to the chain itself, which modifications will produce molecules falling within the scope of the present invention, as long as the resulting molecule possesses surfactant activity as described herein.
  • the employed pharmaceutical composition contains a mixture of a surfactant polypeptide and phospholipid.
  • the composition can contain the KL 4 polypeptide combined with phospholipids dipalmitoyl phosphatidylcholine (DPPC) and palmitoyl- oleoylphosphatidyl glycerol (POPG) at a 3 : 1 ratio, and also palmitic acid.
  • DPPC dipalmitoyl phosphatidylcholine
  • POPG palmitoyl- oleoylphosphatidyl glycerol
  • This KL 4 -Surfactant is being marketed under the name Model surfactant mixture.
  • the efficacy of KL 4 -Surfactant in various experimental and clinical studies has been previously reported, see, e.g.
  • polypeptide:phospholipid weight ratio can be in the range of about 1 :5 to about 1 : 10,000, preferably about 1 :7 to about 1 :5,000, more preferably about 1 : 10 to about 1 : 1 ,000, and most preferably about 1 : 15 to about 1 : 100. In a particular preferred embodiment, the polypeptide:phospholipid weight ratio is about 1 :37.
  • agents compatible with or suitable for treating pulmonary conditions can also be included in the compositions of the invention. These agents include any compounds that are beneficial to treating or alleviating symptoms associated with acute asthma or bronchiolitis.
  • agents that can be co-administered include anti-allergenic agents, anti-inflammatory agents, anti-microbials including anti-bacterials, anti-fungals, and anti-virals, antibiotics,
  • immunomodulators such as hematopoietics, leukotriene modifiers, xanthines, sympathomimetic amines, mucolytics, corticosteroids, anti-histamines, and vitamins.
  • Other examples include bronchodilators, such as albuterol, levalbuterol (e.g., Xopenex®), terbutaline, salmeterol, formoterol, and pharmacologically acceptable salts thereof, anticholinergics, such as ipratropium bromide, the so-called “mast cell stabilizers," such as cromolyn sodium and nedocromil, corticosteroids, such as flunisolide, fluticasone, beclomethasone, budesonide, triamcinolone, and salts thereof, interferons such as INF-alpha, beta and gamma, mucolytics, such as N-acetylcysteine and guaifenesin, le
  • the pharmaceutical compositions of the invention can contain inhibitors of tissue destruction, especially those inhibitors that can minimize tissue damage during inflammation.
  • Suitable inhibitors of tissue destruction for the invention include, e.g., protease inhibitors, anti-oxidants, phospholipase inhibitors, lipase inhibitors and combinations thereof can be used in the methods of the invention. These agents may be in the form of proteins, peptides, nucleic acids, polysaccharides, carbohydrates, lipids, glycoproteins, and organic and inorganic compounds.
  • the invention contemplates use of any available protease inhibitor in the compositions and methods of the invention. These include, for example, the Kunitz inhibitors, matrix metalloproteinase inhibitors and serine protease inhibitors.
  • protease inhibitors include inhibitors of trypsin, chymotrypsin, elastase, kallikrein, plasmin, coagulation factors XIa and IXa, and cathepsin G.
  • Serine proteases include such enzymes as elastase (e.g.
  • inhibitors of human leukocyte elastase cathepsin G, plasmin, C-l esterase, C-3 convertase, urokinase, plasminogen activator, acrosin, chymotrypsin, trypsin, thrombin, factor Xa and kallikreins.
  • Another family of inhibitors includes inhibitors of metalloproteinases such as any of metalloproteinases 1 -13.
  • elastase inhibitors that can be used in the invention include, for example, human leukocyte elastase inhibitor, elafin and alpha- 1 - proteinase inhibitor.
  • suitable protease inhibitors include human secretory leukocyte protease inhibitor, alpha- 1 -antitrypsin, alpha- 1-antichymotrypsin, C- reactive protein and combinations thereof.
  • Phospholipase enzymes catalyze the removal of fatty acid residues from phosphoglycerides. Specifically, phospholipase A2 (PLA2) cleaves the ester bond at the 2 position of the glycerol moiety of membrane phospholipids giving rise to equimolar amounts of arachidonic acid and lysophospholipids. Although PLA2 preferentially cleaves arachidonic acid from phospholipids, arachidonic acid is generated secondarily from intermediates of the SI , phospholipase C- and phospholipase D-activated pathways. PLA2 inhibitors include chemical molecules such as p-bromophenacyl bromide.
  • PLA2 inhibitors include biological molecules such as thielocin A l beta, produced by a fungus (Tanaka et al. (1995) Eur. J. Pharmacol. 279: 143-8), or lipocortin or annexin I (NCBI accession number gi:71756; Wallner et al., Cloning and expression of human lipocortin, a phospholipase A2 inhibitor with potential anti-inflammatory activity, Nature 320 (6057), 77-81 (1986)), or Crotalus phospholipase A2 inhibitor (CNF) (NCBI accession number gi: 501050; Fortes-Dias C L et al. 1 94; J. Biol. Chem. 269: 15646-51 ).
  • Nonspecific PLA2 inhibitors such as glucocorticoids can also be used.
  • Phospholipase A 2 inhibitors suitable for use in the invention also include LY31 1 -727 (Eli Lilly).
  • Anti-oxidants can also be employed in the compositions of the invention. Inflammation can stimulate polymorphonuclear leukocytes and macrophages that produce large amounts of superoxide (0 2 * _ ) and hydrogen peroxide (H 2 0 2 ) (Babior, B. M. et al. [1973] J Clin Invest 52:741-744; Halliwell, B. et al. [1999] Free radicals in Biology and Medicine. Oxford N.Y.: Clarendon Press, Oxford University Press). The detrimental effects of these radicals may be amplified in the presence of iron and the subsequent formation of other reactive
  • Suitable anti-oxidants include catalase, glutathione, N-acetylcysteine, procysteine, rosemary leaf extract, alpha-tocopherol, 2,4-diaminopyrrolo-[2,3-d]pyrimidine, ascorbic acid and carotenoid compounds such as leutein, zeaxanthin, cryptoxanthin, violaxanthin, carotene diol, hydroxycarotene, hydroxylycopene, alloxanthin, ebselen and dehydrocryptoxanthin, including derivatives thereof.
  • ester derivatives of ascorbic acid and of carotenoid compounds such as lutein, zeaxanthin, cryptoxanthin, violaxanthin, carotene diol, hydroxycarotene, hydroxylycopene, alloxanthin and dehydrocryptoxanthin can be used in the invention.
  • spreading agents can be used in the compositions of the invention. While not wishing to be limited to a specific mechanism, the spreading agent is believed to promote transition of surfactant-mixture lipids from particle form to monolayer form, leading to spreading on and distribution along and within the respiratory system (e.g., lung surface). Thus, for example, if the surfactant formulation is delivered to the lung in liposomal form, the spreading agent is effective in promoting transition of the liposomal
  • the spreading agent is effective in promoting transition of the surfactant-mixture phospholipids to a planar monolayer form at the lung surface.
  • Exemplary spreading agents include but are not limited to non- phospholipid lipids that are compatible with lipid bilayer or lipid monolayer formation, but which alone are not able to support lipid-bilayer formation.
  • Exemplary spreading agents include lysophospholipids; fatty acids, fatty esters, and fatty alcohols, and other single-long-chain fatty acyl compounds.
  • Preferred spreading agents include fatty acids and fatty alcohols having alkyl chain lengths of at least about 12 carbon atoms, preferably between 15-20 carbon atoms in chain length.
  • One preferred spreading agent is palmitic acid; another is cetyl alcohol.
  • the spreading agent makes up about 2 to about 25 dry weight percent of the surfactant composition, or about 10 to about 15 dry weight percent of the composition.
  • the spreading agents used in the present invention may be purchased from commercial suppliers.
  • palmitic acid (PA) may be obtained from Avanti Polar Lipids, Inc. (Birmingham, Ala.).
  • the spreading agents may also be prepared according to methods available in the art.
  • the composition can include Tyloxapol as a spreading agent, which can be purchased under several trade names from various companies such as Sterling-Winthrop, and Rohm and Haas.
  • Tyloxapol is a polymer of 4-(l , l ,3,3-tetramethylbutyl)phenol) with formaldehyde and oxirane. Tyloxapol has been used in human pharmacologic formulations for over 30 years (Tainter ML et al. New England Journal of Medicine (1955) 253 :764-767). Tyloxapol is relatively nontoxic and does not hemolyze red blood cells in a thousand times the concentrations at which other detergents are hemolytic (Glassman HN. Science (1950) 1 1 1 :688-689). Treatment methods
  • the invention provides compositions and methods for promoting bronchioli dilatation and alveoli expansion in needing subjects.
  • Treatment methods of the invention typically involve administering to the subject a therapeutically effective amount of a composition comprising a phospholipid or a phospholipid-surfactant polypeptide mixture, as described herein.
  • the pharmaceutical compositions of the invention can be employed alone to treat subjects suffering from or at risk of developing the bronchiole and alveoli related pulmonary condition, e.g., patients suffering from acute asthma or bronchiolitis.
  • the subjects are administered with a composition that contains a mixture of a surfactant polypeptide and phospholipids (e.g., the KL4- Surfactant).
  • the administered composition contains only the phospholipids as the active ingredient.
  • phospholipids administered in the composition can consist of, but are not limited to, depalmitoyl phosphatidylcholine (DPPC) and palmitoyl oleoyl
  • the pharmaceutical composition containing the phospholipids or surfactant polypeptide-phospholipid mixture is administered as an aerosol.
  • a freeze-dried preparation of the composition can be placed into a liquid medium and put into an aerosol by any aerosol device or nebulizer.
  • the phospholipids or the surfactant-phospholipids mixture e.g., KL 4 -Surfactant
  • the phospholipids or the surfactant-phospholipids mixture are prepared in an aqueous emulsion at a concentration that will be the highest possible for aerosolization, e.g., 20-50 mg/ml.
  • the compositions can be administered in the presence of Continuous Positive Airway Pressure (CPAP).
  • CPAP Continuous Positive Airway Pressure
  • the administration of the aerosolized composition can continue along with CPAP until air-trapping has cleared or has been greatly reduced.
  • CPAP will continue to allow the administered composition (e.g., aerosolized phospholipids or L 4 -Surfactant) to stabilize along the surfaces of the bronchioles and alveoli where it forms a molecular layer with lateral stability that prevents collapse of the bronchioles and alveoli.
  • the administration of the composition (phospholipid or KL 4 - Surfactant aerosol) and CPAP can be repeated as needed since the inflammatory process that initiated the disease may require time to clear.
  • CPAP devices are well known and routinely used in the art, and can be readily obtained from many commercial sources. These include, e.g., Philips Respironics (PA, USA),
  • a CPAP machine can be coupled to the delivery device such as a nebulizer (e.g., a Pari LC Star nebulizer as detailed below).
  • a nebulizer e.g., a Pari LC Star nebulizer as detailed below.
  • a T-coupled junction tube can be used that directs both the CPAP air stream and the nebulizer air stream into a single T junction and thereafter directly into the patient's airways for breathing.
  • a CPAP machine can be coupled to a Pari LC Star nebulizer by hooking the junction tube into the air entry at the top of the medication cup of the nebulizer.
  • subjects treated with the methods and compositions described herein can be further examined with a means to measure reduction of air-trapping in the lung.
  • the examination can be performed prior to, simultaneous with or subsequent to treatment with the therapeutic compositions of the invention. This allows one to monitor efficacy of the employed therapy.
  • Air-trapping in the lungs of the patients can be examined with any methods available in the art. For example, air-trapping in the subjects can be quantified with the helium breath technique.
  • the helium dilution technique is a well-known way of measuring the functional residual capacity of the lungs (the volume left in the lungs after normal expiration).
  • the helium spreads into the lungs of the patient,and settles at a new concentration (C2). Because there is no leak of substances in the system, the amount of helium remains constant during the test, and the FRC can then be calculated.
  • the helium dilution test is preferred in the invention to monitor and quantify air-trapping and therapeutic removal of air-trapping in patients with various pulmonary conditions, including subjects with acute bronchiolitis.
  • subjects afflicted with a bronchiole related pulmonary condition such as acute asthma can be treated with the pharmaceutical composition described herein in combination with other known treatment regimens.
  • the composition containing the surfactant can be administered to the subject in combination with other compounds suitable for treating or ameliorating symptoms of the bronchiole and alveoli related pulmonary condition (e.g., bronchiolitis or asthma attack).
  • the treatment can consist of standard of care in which corticosteroids and possible p 2 -adrenergic agonists as well as other anti-inflammatory agents will be administered.
  • the pharmaceutical composition described herein e.g., KL4-Surfactant
  • the pharmaceutical composition can be administered, e.g., as an aerosol.
  • the pharmaceutical composition can be administered to subjects suffering from or afflicted with a bronchiole and alveoli related pulmonary condition (e.g., asthma attack or bronchiolitis) with one or more antibiotic drug to treat bacterial infection.
  • a combination therapy is especially suitable for subjects suspected to have bacterial infection.
  • a pharmaceutical composition of the invention can be combined with known symptom-managing regimen for the disorder, e.g., the use of inhaled epinephrine or hypertonic saline.
  • the pharmaceutical composition can be administered to the patients prior to, concurrently with, or subsequent to treatment with the known therapy.
  • any mammalian subjects suffering from a bronchiole and alveoli related pulmonary condition such as acute asthma and/or bronchiolitis can be treated with the compositions and methods of the invention.
  • the subjects are human patients.
  • adults, teenagers, children, infants and pre-term infants can all be treated with the compositions and methods of the invention.
  • the treatment should result in improvement in one or more symptoms in the patients associated with the condition (decreased inflammation, expanded aveoli or dilated bronchioles).
  • the phospholipids or phospholipid-surfactant polypeptide mixture described herein can be formulated into a variety of acceptable formulations or pharmaceutical compositions.
  • the compositions can additional contain other compounds described herein, e.g., spreading agents.
  • Therapeutic compositions of the invention may also contain a physiologically tolerable or acceptable carrier or excipient, dissolved or dispersed therein.
  • the compositions can include protease inhibitors, lipase inhibitors, anti-oxidants, as well as other therapeutic agents known to be helpful to treating symptoms associated with bronchiole and alveoli related pulmonary conditions (e.g., as anti-inflammatory agents or anti-allergy agents).
  • the therapeutic composition is not immunogenic when administered to a mammal or human patient for therapeutic purposes.
  • compositions can be administered to a mammalian host, such as a human patient, in a variety of forms adapted to the chosen route of administration, e.g., aerosol administration via the tracheo-bronchial route.
  • aerosol administration the only structural requirement of the pharmaceutical formulation is that be it can be converted or processed into a suitable aerosol- particle form containing the components of the composition.
  • the composition can be an aqueous suspension of lipid bodies prepared by lyophilization to form a dry mass that is then comminuted, e.g., by grinding, to form a composition containing dry-powder particles having a mass median aerodynamic diameter in the 1 -5 ⁇ size range.
  • the dry-powder particles are then stored and employed in a suitable aerosolization device to produce a dry-particle aerosol suitable for inhalation treatment or for suspension in a suitable solvent, for aerosolization as a particle suspension.
  • some therapeutic methods of the invention employ pharmaceutical compositions that contain only phospholipids without any polypeptide ingredient.
  • the phospholipid (PL) upon administered to the patients can form functional lung surfactant "in situ" by association with native SP-B protein secreted by epithelial cells in the respiratory tissues.
  • the administered pharmaceutical composition comprise respirable particles that can be delivered to bronchi and bronchioles. Typically, these particles have a mass median aerodynamic diameter ranging from about 5 ⁇ to about 0.5 ⁇ or about 0.3 ⁇ .
  • the administered pharmaceutical composition comprise primarily respirable particles that can reach bronchioles. These methods are particularly suitable for treating bronchiolitis by reducing collapse of the bronchioles.
  • the invention contemplates processing a liquid formation by means of a user-controlled nebulizer or aerosolizer, to generate an aqueous- droplet aerosol containing the phospholipid or phospholipid-surfactant formulation in lipid-body form.
  • the phospholipid and the surfactant components of the formulation can be present in ordered, crystalline, or amorphous lipid particles suspended in the aerosol droplets.
  • some embodiments of the invention employ a Pari Star nebulizer (PARI Respiratory Equipment, Inc., VA, USA) for administering the therapeutic compositions of the invention to patients in need of treatment.
  • a Pari LC Star nebulizer can be used for administering
  • compositions containing the respirable particles described above This device is designed to target peripheral airways and lung parenchyma (tissue). It allows delivery of a high percentage of particles of less than 5 ⁇ in size, esp. for delivery of particles sized around 1 ⁇ to 2 ⁇ range to the near terminal and terminal bronchioles. Some other methods can use the Salter 8900 series nebulizer (Salt Labs, CA, USA). This device can deliver more than 60% of the particles that are less than 2 ⁇ in size.
  • a nebulizer mask instead of a nebulizer can be used for administering the therapeutic compositions of the invention, e.g., in the treatment of children or infants.
  • Bubbles The FishTM II pediatric masks are suitable for delivering the most respirable particle medication while reducing facial and eye deposition.
  • the formulation is processed by spray drying to produce spray-dried particles having the desired mass median aerodynamic diameter in the 1 -5 ⁇ size.
  • the spray dried particles may then be stored and employed by the user in an aerosolization device, as above, for inhalation therapy.
  • the powdered particles can be delivered as a dry-powder aerosol, or the particles can be suspended in an aqueous medium for aerosolization in aqueous droplet form.
  • a suitable surfactant formulation in liquid form e.g., a formulation solution or suspension contained in a volatile biocompatible fluid, may be formed in an aerosolization process in which the particles formed are immediately inhaled for therapeutic delivery of the active agent.
  • Freeze drying is one standard method for producing a dry powder from a solution or a suspension. See, for example, Freide, M., et al, Anal. Biochem., 21 1 (1): 1 17-122, 1993; Sarbolouki, M.N. and T. Toliat, PDA J. Pharm. Sci. Technol, 52(l):23-27, 1998).
  • the dried surfactant formulation is comminuted, e.g., by grinding or other conventional means, to form desired size particles.
  • Spray drying may also be used advantageously for producing dried lipid particles of desired sizes.
  • Various spray-drying methods have been described in the patent literature, See, for example, U.S. Patent Nos. 6, 174,496; 5,976,574; 5,985,284; 6,001 ,336; 6,015,256; 5,993,805 ; 6,223,455; 6,284,282; and 6,051 ,257.
  • the amount of the therapeutic composition administered to a patient can be in the range of about 0.5-2,500 mg/dose, 1 - 1 ,500 mg/dose, 2-500 mg/dose, 5-100 mg/dose, or 10-50 mg/dose. Adjustments to the dose, to optimize therapeutic effectiveness, and minimize side effects, can be determined according to known procedures.
  • the active phospholipid or surfactant polypeptide present in formulation e.g., an aerosol
  • duration of treatment will be continuous until symptoms of the bronchiole and alveoli related pulmonary condition (e.g., asthma attack) are gone or controlled.
  • the therapeutic formulation can be administered to the subjects about once a day, twice a day, four times a day, six times a day, once every hour, once every half hour, once every 10 minutes or even more frequently.
  • the patients can be administered 5-50 mg aerosolized KL4-Surfactant or the phospholipids DPPC and POPG (with or without palmitic acid) every 5, 10, 15, 20, 25, or 30 minutes. In other embodiments, the administration can take place once every hour, every 2, 4, 6, 8 hours or even longer.
  • patients can receive the aerosolized KL4-Surfactant or phospholipids during a period of, e.g., 5-60 minutes, depending on the severity of their condition.
  • the aerosol can be administered with a synchronized pressure system, adding pressure during inspiration and maintaining positive airway pressure (CPAP) upon expiration.
  • CPAP positive airway pressure
  • the pressure of CPAP applied during the treatment will be in the range of about 4 cm H 2 0 to about 20 cm H 2 0.
  • the exact degree of inspiratory and expiratory pressure will depend upon the patient's condition (such as weight and other medical factors), and can be accordingly adjusted by referring to the user manual of the CPAP machine employed, as well as knowledge well known in the art about CPAP (see, e.g., Berry et al., Sleep, 25 : 148-173, 2002 ).
  • the applied CPAP pressure will usually decrease as the patient's symptoms improve.
  • the purpose of applying CPAP during and/or after administering the formulation is to open the bronchioles and alveoli and maintain them in the expanded position so as to allow the KL4-Surfactant or phospholipids to form a monolayer with resulting expansion.
  • the entire cycle of treatment can be repeated after one hour if the patient's condition worsens.
  • a less frequent treatment regimen may be employed.
  • a less frequent administration may also be appropriate.
  • some subjects may be administered the pharmaceutical composition less frequently, e.g., once an hour, once every 4 hours, once a day, once every second, third to sixth day, once a week or even less frequently.
  • the therapeutic formulation may be administered to a subject at varying frequency during the treatment period.
  • Synthesis of a surfactant polypeptide of the present invention may be carried out according to a variety of known methods of synthesis. The following procedure is described as exemplary. Chemicals and reagents useful in synthesizing batches of surfactant peptides, e.g., batches of KL4 peptide, include the following:
  • DCM Dichloromethane
  • Trifluoroacetic acid (TFA; Halocarbon);
  • DIEA Diisopropylethylamine
  • NMP N-Methylpyrrolidone
  • Acetic anhydride (Ac 2 0; Mallinckrodt, St. Louis, MO);
  • L 4 peptide SEQ ID NO: l
  • Coupler 296 Peptide Synthesizer Vega Biotechnologies, Arlington, AZ
  • the appropriate amino acid derivative was being made.
  • the appropriate amino acid was dissolved in one ( 1 ) liter of NMP. After a clear solution was obtained, HOBt was added to the solution. When the HOBt was dissolved, DCC was added to the solution. This solution was left stirring for one (1) hour at room temperature. During this one hour of stirring, a by-product formed, dicyclohexylurea (a white precipitate). This by-product was filtered off through a buchner funnel using Whatman's #1 filter paper. The filtrate was then added manually to the contents of the Vega 296 reaction vessel at step No. 7.
  • the synthesizer was then programmed to stop after the completion of step No. 9. Aliquots of the peptide resin were subjected to the quantitative ninhydrin test of Sarin et al. (Applied Biosystems 431 A user manual, Appendix A). The coupling efficiencies were good throughout the entire synthesis.
  • the unreacted peptide resin was acetylated after leucine 12 (cycle 9) and after leucine 5 (cycle 16). After each acetylation, the peptide resin was washed with dichloromethane (see Table 2, step 1 1).
  • the completed peptide resin was deprotected (removal of the Boc group) by completing steps 1 -3 of the program (see Table 2).
  • the deprotected peptide resin was then washed with ample volumes of absolute ethanol and dried in vacuo over P2O5
  • the weight of the dried, deprotected peptide resin was 256.48 grams. Since the batch was started with 100 g of t-Boc-Lysine (Cl-Z) OCH 2 PAM resin at a substitution of 0.64 mmoles/gram, the load corresponded to 64 mmoles. Subtracting out the initial 100 grams of resin, the weight gain was 156.48 grams.
  • the molecular weight of the nascent protected peptide (excluding the C-terminal lysine anchored onto the resin) was 301 1.604 g/mole.
  • the HF apparatus was then equilibrated to atmospheric pressure and an oily sludge was found at the bottom of the reaction vessel.
  • Cold anhydrous ether 700 ml, prechilled to -20°C
  • the resin clumps were triturated with ether using a glass rod.
  • the ether was decanted after the resin settled.
  • the resin was then washed with 500 ml of room temperature anhydrous ether and allowed to stir for about 5 min.
  • the ether was decanted after the resin settled.
  • the resin was washed until it became free- flowing (4-5 total washes). The resin was left in the fume hood to dry overnight.
  • the resulting dried HF-treated resin was then weighed and stored in the freezer. 1.021 grams of the dried HF-treated resin was removed and extracted with 50 ml of 50% acetic acid/water and allowed to stir for 30 min. The resin was filtered through a coarse sintered glass funnel, and the filtrate was collected in a lyophilizing jar. The filtrate was diluted with approximately 200 ml of water, shell frozen, and placed on the lyophilizer. The one (1) gram of extracted HF-treated resin yielded 569 mg of crude peptide. The following table (Table 3) summarizes the large scale HF treatments of the remaining KL4 peptide resin. All of the HF-treated resins were stored in the freezer.
  • the peptide was purified using a Dorr-Oliver Model B preparative HPLC (Dorr-Oliver, Inc., Milford, CT). This unit was connected to a Linear Model 204 spectrophotometer and Kipp and Zonen dual channel recorder. This preparative HPLC was interfaced with a Waters KL250 Column Module (Waters Associates, Milford, MA) containing a radially compressed 10x60 cm cartridge filled with Vydac C4 support, 15-20 microns, and 300 A pore size (Vydac, Hesperia, CA).
  • Solvent "A” consisted of 0.1% HOAc in water
  • solvent “B” consisted of 0.1% HOAc in acetonitrile.
  • the flow rate was set at 400 ml/min
  • the cartridge was compressed to 150-200 psi
  • the preparative HPLC system back pressure was at 550-600 psi.
  • BPS # 1 The total volume of BPS # 1 was approximately 60 liters. Reverse osmosis was used to concentrate the peptide solution to a final volume of two liters. A Millipore Model 6015 Reverse osmosis Unit with an R75A membrane to retain the peptide was used. The resulting two liters of BPS #1 were filtered through a buchner funnel using two pieces of Whatman #1 filter paper, divided into approximately 1 1 lyophilizing jars and diluted with equal volumes of water. The lyophilizing jars were shell- frozen and lyophilized. The total weight of dry KL 4 peptide at the end of the procedure was 40.25g.
  • a stock solution of surfactant composition was formulated to contain 40 mg/mL total phospholipid, with a composition based on the following formula:
  • surfactant compositions were made that contained varying amounts of palmitic acid (PA) and the KL 4 peptide in 2.5 to 30 mg per mL of total phospholipids (Table 4).
  • a Model Surfactant Mixture was made as follows. KL 4 peptide (9 mg), DPPC (225 mg), POPG (75 mg) and PA (45 mg) were dissolved in 2.5 milliliters (ml) of 95% ethanol at 45°C. This solution was then added to 7.5 ml of distilled H 2 0 at 45°C with rapid vortexing and 2 ml of 500 mM NaCl, 250 mM Tris- acetate pH 7.2 was added. The resulting milky suspension was stirred at 37°C for 15 minutes and the ethanol present was then removed by dialysis (Spectrapor 2; 13,000 mol. wt. cutoff) against 100 volumes of 130 mM NaCl, 20 mM Tris- acetate pH 7.2 buffer at 37°C. Dialysis was continued for 48 hours with two changes of the dialysis solution.
  • composition may further comprise a buffer
  • Glacial acetic acid, USP quantity sufficient to adjust
  • Tham buffered system was prepared essentially as follows. 0.37 ml of Tham solution (tromethamine injection, NDC 0074-1593-04, Abbott
  • an antibody includes a plurality (for example, a solution of antibodies or a series of antibody preparations) of such antibodies, and so forth.
  • a reference to “an antibody” includes a plurality (for example, a solution of antibodies or a series of antibody preparations) of such antibodies, and so forth.
  • the patent be interpreted to be limited to the specific examples or embodiments or methods specifically disclosed herein.
  • the patent be interpreted to be limited by any statement made by any Examiner or any other official or employee of the Patent and Trademark Office unless such statement is specifically and without qualification or reservation expressly adopted in a responsive writing by Applicants.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Medicinal Chemistry (AREA)
  • Veterinary Medicine (AREA)
  • General Health & Medical Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Engineering & Computer Science (AREA)
  • Animal Behavior & Ethology (AREA)
  • Pulmonology (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Dispersion Chemistry (AREA)
  • Otolaryngology (AREA)
  • Medicinal Preparation (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention concerne des compositions et méthodes pour traiter et prévenir les affections pulmonaires avec dilatation des bronchioles dysfonctionnels telle que l'asthme et la bronchiolite. Les méthodes utilisent une formulation contenant des phospholipides seuls ou des phospholipides associés à un polypeptide tensioactif décrit dans l'invention pour améliorer la dilatation de bronchioles et éventuellement la dilatation d'alvéoles atélectasiques chez les patients.
PCT/US2014/061717 2013-10-22 2014-10-22 Méthodes et compositions pour favoriser la dilatation des bronchioles Ceased WO2015061412A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201361894125P 2013-10-22 2013-10-22
US61/894,125 2013-10-22

Publications (1)

Publication Number Publication Date
WO2015061412A1 true WO2015061412A1 (fr) 2015-04-30

Family

ID=52993486

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2014/061717 Ceased WO2015061412A1 (fr) 2013-10-22 2014-10-22 Méthodes et compositions pour favoriser la dilatation des bronchioles

Country Status (1)

Country Link
WO (1) WO2015061412A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2611406C1 (ru) * 2015-09-03 2017-02-21 Общество с ограниченной ответственностью "Биосурф" (ООО "Биосурф") Способ лечения бронхиальной астмы

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5407914A (en) * 1988-01-06 1995-04-18 The Scripps Research Institute Pulmonary surfactant protein and related polypeptides
US20050070477A1 (en) * 2002-04-25 2005-03-31 The Scripps Research Institute Treatment and prevention of pulmonary conditions
US7101341B2 (en) * 2003-04-15 2006-09-05 Ross Tsukashima Respiratory monitoring, diagnostic and therapeutic system
US20080199410A1 (en) * 2006-09-19 2008-08-21 Discovery Laboratories, Inc. Pulmonary surfactant formulations and methods for promoting mucus clearance

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5407914A (en) * 1988-01-06 1995-04-18 The Scripps Research Institute Pulmonary surfactant protein and related polypeptides
US20050070477A1 (en) * 2002-04-25 2005-03-31 The Scripps Research Institute Treatment and prevention of pulmonary conditions
US7101341B2 (en) * 2003-04-15 2006-09-05 Ross Tsukashima Respiratory monitoring, diagnostic and therapeutic system
US20080199410A1 (en) * 2006-09-19 2008-08-21 Discovery Laboratories, Inc. Pulmonary surfactant formulations and methods for promoting mucus clearance

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2611406C1 (ru) * 2015-09-03 2017-02-21 Общество с ограниченной ответственностью "Биосурф" (ООО "Биосурф") Способ лечения бронхиальной астмы

Similar Documents

Publication Publication Date Title
AU2003223718B2 (en) Treatment and prevention of pulmonary conditions
US8337815B2 (en) Pulmonary surfactant formulations
JP5575639B2 (ja) 改善された特性を有する再構成サーファクタント
TWI388333B (zh) 具有改良性質之重組表面張力素
JP2013177422A (ja) サーファクタント処置療法
US20070129297A1 (en) Treatment and prevention of asthma
WO2007005672A2 (fr) Traitement et prevention de maladies et de situations respiratoires
WO2015061412A1 (fr) Méthodes et compositions pour favoriser la dilatation des bronchioles
HK1109077B (en) Pulmonary surfactant formulations

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 14855977

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 14855977

Country of ref document: EP

Kind code of ref document: A1