WO2014098232A1 - Pharmaceutical composition comprising pi3 kinase inhibitor, pharmaceutical composition comprising compound which acts on vitamin d receptor, freeze-dried composition, method for producing freeze-dried composition, and pharmaceutical composition for transpulmonary administration - Google Patents

Pharmaceutical composition comprising pi3 kinase inhibitor, pharmaceutical composition comprising compound which acts on vitamin d receptor, freeze-dried composition, method for producing freeze-dried composition, and pharmaceutical composition for transpulmonary administration Download PDF

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WO2014098232A1
WO2014098232A1 PCT/JP2013/084310 JP2013084310W WO2014098232A1 WO 2014098232 A1 WO2014098232 A1 WO 2014098232A1 JP 2013084310 W JP2013084310 W JP 2013084310W WO 2014098232 A1 WO2014098232 A1 WO 2014098232A1
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group
freeze
composition
pharmaceutical composition
vitamin
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PCT/JP2013/084310
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French (fr)
Japanese (ja)
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親正 山下
堀口　道子
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Yamashita Chikamasa
Horiguchi Michiko
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    • 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
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
    • A61K31/203—Retinoic acids ; Salts thereof
    • 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
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • 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
    • A61K31/59—Compounds containing 9, 10- seco- cyclopenta[a]hydrophenanthrene ring systems
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers

Definitions

  • the present invention relates to a pharmaceutical composition containing a PI3 kinase inhibitor, a pharmaceutical composition containing a compound acting on a vitamin D receptor, a lyophilized composition for producing a pharmaceutical composition for pulmonary administration, and the lyophilized composition And a pharmaceutical composition for pulmonary administration.
  • COPD chronic obstructive pulmonary disease
  • pulmonary emphysema chronic bronchitis
  • acute lung distress syndrome (ARDS) acute lung distress syndrome
  • pulmonary fibrosis lung cancer
  • stroma Pneumonia pulmonary tuberculosis sequelae
  • pneumoconiosis etc.
  • COPD chronic obstructive pulmonary disease
  • ARDS acute lung distress syndrome
  • pulmonary fibrosis lung cancer
  • stroma Pneumonia pulmonary tuberculosis sequelae
  • pneumoconiosis etc.
  • a therapeutic agent that induces differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells constituting the alveoli and regenerates the alveoli is considered promising. It has been.
  • an inhalation system for powder medicine for pulmonary administration there is an inhalation system that particles a freeze-dried composition by air impact (see, for example, US Pat. No. 7,735,485 B2 and US Pat. No. 4,822,709).
  • the freeze-dried composition becomes particles suitable for transpulmonary administration by inhalation of the patient, and inhalation is performed as it is to perform transpulmonary administration.
  • this system uses a lyophilization method suitable for water-soluble drugs, such as proteins and peptides, and is suitable for transpulmonary formulation of drugs that are poorly soluble in water such as retinoic acid. Not. Therefore, in order to apply this system to poorly soluble drugs such as retinoic acid, it is necessary to find a new formulation and manufacturing method.
  • a compound that induces differentiation of alveolar epithelial stem cells into alveolar epithelial cells has been found and the compound can be formulated into a pharmaceutical composition for transpulmonary administration, regeneration of alveoli will be possible for diseases exhibiting lung tissue damage. It is possible to provide a treatment method having an action point.
  • a mode of administration of a pharmaceutical composition for pulmonary administration if the powder drug inhalation system for pulmonary administration disclosed in US Pat. No. 7,735,485B2 or US Pat. A regenerative therapy for alveoli can be provided simply.
  • the present invention was made under the above situation.
  • a differentiation inducer that acts on alveolar epithelial stem cells comprising a PI3 kinase inhibitor as an active ingredient.
  • a pharmaceutical composition for inducing alveolar regeneration comprising a PI3 kinase inhibitor as an active ingredient.
  • a pharmaceutical composition for treating lung tissue injury comprising a PI3 kinase inhibitor as an active ingredient.
  • A5 Use of a PI3 kinase inhibitor in the manufacture of a pharmaceutical composition that induces alveolar regeneration.
  • A6 Use of a PI3 kinase inhibitor in the manufacture of a pharmaceutical composition for treating lung tissue damage.
  • PI3 kinase inhibitor for use in the treatment of lung tissue damage.
  • a method for treating lung tissue damage comprising administering a pharmaceutical composition comprising a PI3 kinase inhibitor as an active ingredient to a patient having damage to lung tissue.
  • the PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- PI3, an agent that inhibits the p40phox pathway Agent
  • a differentiation inducer that acts on alveolar epithelial stem cells comprising a compound that acts on a vitamin D receptor as an active ingredient.
  • B2 A pharmaceutical composition for inducing alveolar regeneration, comprising a compound that acts on a vitamin D receptor as an active ingredient.
  • B3 A pharmaceutical composition for treating lung tissue damage, comprising a compound that acts on a vitamin D receptor as an active ingredient.
  • B4 Use of a compound that acts on a vitamin D receptor in the production of a differentiation inducer that acts on alveolar epithelial stem cells.
  • B5 Use of a compound that acts on a vitamin D receptor in the manufacture of a pharmaceutical composition that induces alveolar regeneration.
  • (B6) Use of a compound that acts on a vitamin D receptor in the manufacture of a pharmaceutical composition for treating lung tissue damage.
  • a method for treating lung tissue damage comprising administering to a patient having damage to lung tissue a pharmaceutical composition comprising a compound that acts on a vitamin D receptor as an active ingredient.
  • the compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and derivatives thereof, wherein (B1) ⁇ (B8).
  • (C1) Freeze-drying that contains a poorly water-soluble drug, at least two types of amino acids, and a surfactant, and the proportion of particles having a particle diameter of 5 ⁇ m or less becomes 10% or more when formed into particles by air flow Composition.
  • (C101) contains a poorly water-soluble drug, at least two types of amino acids, and a surfactant, and when formed into particles by an air stream, the proportion of particles having a geometric particle diameter of 5 ⁇ m or less is 10% or more.
  • a lyophilized composition is a poorly water-soluble drug, at least two types of amino acids, and a surfactant
  • (C102) contains a poorly water-soluble drug, at least two kinds of amino acids, and a surfactant, and when formed into particles by an air stream, the proportion of particles having an aerodynamic particle diameter of 5 ⁇ m or less is 10% or more.
  • a lyophilized composition contains a poorly water-soluble drug, at least two kinds of amino acids, and a surfactant, and when formed into particles by an air stream, the proportion of particles having an aerodynamic particle diameter of 5 ⁇ m or less is 10% or more.
  • C2 A lyophilized composition of a composition comprising a drug that is sparingly soluble in water, at least two types of amino acids, a surfactant, an alcohol, and water.
  • C201 A freeze-dried composition of a composition comprising a poorly water-soluble drug, at least two types of amino acids, a surfactant, an alcohol, and water, A freeze-dried composition in which the proportion of particles having a particle size of 5 ⁇ m or less is 10% or more.
  • C202 A freeze-dried composition of a composition comprising a poorly water-soluble drug, at least two types of amino acids, a surfactant, an alcohol, and water, A freeze-dried composition in which the proportion of particles having a mechanical particle size of 5 ⁇ m or less is 10% or more.
  • C3 A freeze-dried composition comprising a poorly water-soluble drug, at least two amino acids selected from the following first group, second group and third group and a plurality of groups, and a surfactant: .
  • -Group 1 Phenylalanine, tryptophan-Group 2: Isoleucine, leucine, valine-Group 3: Alanine, glycine
  • C4 Drugs poorly soluble in water, and the following groups 1, 2, 3
  • ⁇ Group 1 phenylalanine, tryptophan
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • the (C5) The (C1) to (C4), (C101), (C102), (C201), and (C1), wherein the water-insoluble drug is a compound having a differentiation-inducing action on alveolar epithelial stem cells.
  • the compound having a differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors
  • the freeze-dried composition according to (C5) which is a seed.
  • the compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and its derivatives, freeze-dried composition according to the (C6).
  • the PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- an agent that inhibits the p40phox pathway, P Agent
  • (C7) A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, the first liquid, at least two kinds of amino acids, a surfactant, water, A method for producing a lyophilized composition comprising the steps of preparing a second liquid by mixing and lyophilizing the second liquid.
  • (C701) A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, and after mixing the first liquid and the surfactant, at least two kinds of A method for producing a lyophilized composition comprising a step of preparing a second liquid by mixing an amino acid and water, and a step of lyophilizing the second liquid.
  • a step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, the first liquid, and the following first group, second group, and third group Freezing comprising a step of preparing a second liquid by mixing at least two kinds of amino acids selected over a plurality of groups, a surfactant, and water, and a step of freeze-drying the second liquid A method for producing a dry composition.
  • -Group 1 Phenylalanine, tryptophan-Group 2: Isoleucine, leucine, valine-Group 3: Alanine, glycine
  • a first liquid is prepared by dissolving a poorly water-soluble drug in a solvent containing alcohol.
  • the compound having a differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors
  • the method for producing a lyophilized composition according to (C9), which is a seed which is a seed.
  • the compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and its derivatives, method for producing a freeze-dried composition according to the (C10).
  • the PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- an agent that inhibits the p40phox pathway, P Production
  • a differentiation inducer that acts on alveolar epithelial stem cells.
  • the present invention also provides a pharmaceutical composition for inducing alveolar regeneration and a pharmaceutical composition for treating lung tissue damage.
  • a freeze-dried composition containing a drug that is sparingly soluble in water, and is formed into particles having a particle size suitable for transpulmonary administration by airflow.
  • a freeze-dried composition used for the production of a pharmaceutical composition for inducing regeneration of alveoli the freeze-dried composition that is granulated into particles having a particle size suitable for transpulmonary administration by air flow Things are provided.
  • a pharmaceutical composition suitable for pulmonary administration which contains a drug hardly soluble in water.
  • the present invention also provides a pharmaceutical composition that induces alveolar regeneration suitable for transpulmonary administration.
  • 2 is a graph showing HE-stained images of a section of lung tissue treated with 1,25-dihydroxyvitamin D 3 and the average distance between alveolar walls in the COPD model mouse of Example B3.
  • a numerical range indicated by using “to” indicates a range including the numerical values described before and after “to” as the minimum value and the maximum value, respectively.
  • the term “process” is not limited to an independent process, and is included in this term if the intended purpose of the process is achieved even when it cannot be clearly distinguished from other processes.
  • the “treatment” may be any improvement of symptoms, and the term also includes suppression of severity and reduction or alleviation of symptoms.
  • “regeneration” is used in the meaning normally used in this technical field.
  • composition containing PI3 kinase inhibitor PI3 kinase (phosphatidylinositol-3 kinase, PI3K) is an enzyme that phosphorylates the hydroxyl group (—OH group) at the 3-position of the inositol ring of inositol phospholipid.
  • Inositol phospholipids are one of the components of eukaryotic cell membranes, and are catalyzed by kinases (phosphorylating enzymes) such as PI3K, so that phosphatidylinositol 3,4,5-triphosphate (PtdIns (3 , 4, 5) P3), and activation of protein kinase B (PKB) / Akt is associated with the control of cell survival, proliferation, differentiation, and cell death.
  • kinases phosphorylating enzymes
  • PtdIns phosphatidylinositol 3,4,5-triphosphate
  • PBB protein kinase B
  • a PI3K inhibitor is an agent that directly inhibits PI3 kinase and an agent that inhibits a downstream signal pathway of PI3K.
  • the downstream signal pathway of PI3K is, for example, PI3K-Akt pathway, PI3K-BTK pathway, PI3K-ITK pathway, PI3K-TAPP pathway, PI3K-DAPP pathway, PI3K-GAB1 / 2 pathway, PI3K-Rac pathway, PI3K-Rho pathway , PI3K-ARF pathway, PI3K-PDK1 pathway, PI3K-EEA1 pathway, PI3K-Rabenosyn5 pathway, PI3K-Rabip4 pathway, PI3K-FAB1 pathway, PI3K-p40phox pathway, PI3K-SNX3 pathway, PI3K-SNX17 pathway, PI3K-SGK pathway It is.
  • PI3K inhibitors have a differentiation-inducing action on alveolar epithelial stem cells.
  • the present inventors have obtained a new finding that PI3K inhibitors induce differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells.
  • PI3K inhibitor examples include the following compounds.
  • Direct inhibitors of PI3K for example, Wortmannin, LY294002, AS605240, ZSTK474, PIK-75 Hydrochloride, IPI-145 (INK1197), GDC-0941, CAL-101 (Idelalisab, GS-1101), BEZ235 NVP-BEZ235, Dactolisib), BKM120 (NVP-BKM120, Buparlisb), GSK2636761, CZC24832, GDC-0032, VS-5684 (SB2343), TG100713, BYL719, CUDC-907, T ), BAY 80-6946 (Copanlist), PF-046 1502, PKI-402, CH5132799, GDC-0980 (RG7422), NU7441 (KU-57788), AS-252424, AS-604850, CAY10505, GSK212645 (GSK458), A66, PF-05212384 (PK
  • a differentiation inducer that acts on alveolar epithelial stem cells, comprising a PI3K inhibitor as an active ingredient.
  • the differentiation inducer differentiates alveolar epithelial stem cells into alveolar epithelial cells.
  • the differentiation inducer contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a pharmaceutical composition for inducing alveolar regeneration comprising a PI3K inhibitor as an active ingredient.
  • the pharmaceutical composition contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
  • the form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a pharmaceutical composition for treating lung tissue damage comprising a PI3K inhibitor as an active ingredient.
  • the pharmaceutical composition contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
  • the form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a method for treating lung tissue damage comprising administering a pharmaceutical composition comprising a PI3K inhibitor as an active ingredient to a patient having damage to lung tissue.
  • the pharmaceutical composition used for the treatment method is the pharmaceutical composition of [2] or the pharmaceutical composition of [3].
  • patients having damage to lung tissue include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, Examples include interstitial pneumonia, pulmonary tuberculosis sequelae, and pneumoconiosis.
  • COPD chronic obstructive pulmonary disease
  • ARDS acute lung distress syndrome
  • pulmonary fibrosis lung cancer
  • Examples include interstitial pneumonia, pulmonary tuberculosis sequelae, and pneumoconiosis.
  • the administration mode is not particularly limited, and various administration routes such as pulmonary administration, intravenous administration, and oral administration can be selected, and preferably pulmonary administration.
  • the dose is selected depending on the type of disease; patient symptom, body weight, age; mode of administration; type of PI3K inhibitor;
  • [5] to [8] and [5 '] to [8'] are provided.
  • [5] to [8] and [5 ′] to [8 ′] correspond to [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration] described later.
  • a PI3K inhibitor as an active ingredient.
  • the details of [5] to [8] and [5 ′] to [8 ′] are described later in [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration]. It is the same.
  • a lyophilized composition comprising a PI3K inhibitor, at least two types of amino acids, and a surfactant.
  • a lyophilized composition comprising a PI3K inhibitor, at least two kinds of amino acids selected from the following first group, second group and third group and a plurality of groups, and a surfactant.
  • ⁇ Group 1 phenylalanine, tryptophan
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • a lyophilized composition comprising a PI3K inhibitor, at least two amino acids, a surfactant, an alcohol, and water.
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • a method for producing a lyophilized composition comprising the steps of preparing a second liquid and lyophilizing the second liquid.
  • [7 ′] A step of preparing a first liquid by dissolving a PI3K inhibitor in a solvent containing alcohol; the first liquid; a plurality of groups from the following first group, second group, and third group: A step of preparing a second liquid by mixing at least two kinds of amino acids selected over the two, a surfactant and water, and a step of freeze-drying the second liquid.
  • a method for producing the composition ⁇ Group 1: phenylalanine, tryptophan ⁇ Group 2: isoleucine, leucine, valine ⁇ Group 3: alanine, glycine
  • a pharmaceutical composition for transpulmonary administration wherein the freeze-dried composition according to [5] or [6] is granulated by airflow.
  • [8 '] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [5'] or [6 '] is granulated by an air stream.
  • a method for transpulmonary administration of a pharmaceutical composition for pulmonary administration comprising the step of preparing the pharmaceutical composition for pulmonary administration by granulating with air impact.
  • a differentiation inducer that acts on alveolar epithelial stem cells, comprising a PI3K inhibitor as an active ingredient; a pharmaceutical composition that induces alveolar regeneration
  • a pharmaceutical composition for treating lung tissue damage a method for treating lung tissue damage; a lyophilized composition used in the manufacture of a pharmaceutical composition for treating lung tissue damage; suitable for transpulmonary administration by airflow;
  • a freeze-dried composition that is granulated into particles having different particle sizes; a method for producing the freeze-dried composition; a pharmaceutical composition for treating lung tissue damage suitable for transpulmonary administration; a pharmaceutical composition for treating lung tissue damage A method of transpulmonary administration of the product.
  • composition containing a compound that acts on vitamin D receptor 1,25-dihydroxyvitamin D 3 (calcitriol) is expressed in lung tissue of patients with chronic obstructive pulmonary disease (COPD) (Sundar IK. Et. Al., Biochem Biophys Res Commun. (2011) 406: 127-133.), But it has been reported to have differentiation-inducing action on alveolar epithelial stem cells. Absent. The present inventors have obtained a new finding that 1,25-dihydroxyvitamin D 3 induces differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells.
  • COPD chronic obstructive pulmonary disease
  • VDR acting compound a compound that acts on VDR (hereinafter referred to as “VDR acting compound”) is used for alveolar epithelial stem cells. It was considered to have a differentiation-inducing action.
  • VDR acting compounds include compounds that bind directly to VDR, eg, VDR ligands, VDR agonists.
  • VDR acting compounds include compounds that do not directly bind to VDR, such as anti-VDR ligand antibodies, compounds that promote signal transduction downstream of VDR, compounds that act on proteins constituting signal transduction pathways downstream of VDR, VDR Compounds that act as activators, compounds that inhibit VDR activator inhibitors are included.
  • VDR ligands and agonists include, for example, vitamin D and derivatives thereof, and specifically include the following compounds.
  • Vitamin D 2 and its derivatives eg ergosterol, ergocalciferol Vitamin D 3 and its derivatives: eg 7-dehydrocholesterol, previtamin D 3 , cholecalciferol, 25-hydroxycholecalciferol, 1,25 Dihydroxycholecalciferol (calcidiol), 1,25-dihydroxyvitamin D 3 (calcitriol), calcitronic acid Vitamin D 4 and its derivatives: eg dihydroergocalciferol Vitamin D 5 and its derivatives Vitamin D 6 And its derivatives Vitamin D 7 and its derivatives Vitamin D analogs: for example, dihydrotaxosterol, calcipotriol, tacalcitol, paricalcitol
  • VDR acting compounds compounds acting on proteins constituting a signal transduction pathway downstream of VDR include, for example, RANKL (Receptor activator of NF-kappa B ligand) inhibitor and NF-kB (Nuclear factor-kappa B) inhibition.
  • RANKL Receptor activator of NF-kappa B ligand
  • NF-kB Nuclear factor-kappa B
  • -RANKL inhibitor For example, WP9QY, Osteoprotegerin (OPG), anti-RANKL antibody (denosumab), RANKL siRNA, RANKL shRNA NF-kB inhibitors: for example, (5Z) -7-Oxoeaenol, (5Z) -Zeaenol, Andrographolide, Aurothiomalate, Capsaicin, (E) -Capsaicin, Evodiamine, anti-NF-kBsRNA, NF-kBsRNA, NF-kBsRNA, NF-kBsRNA
  • the VDR active compound preferably 1,25-dihydroxyvitamin D 3.
  • 1,25-dihydroxyvitamin D 3 shows the differentiation inducing effect on significantly alveolar epithelial stem cells at lower concentrations than ATRA.
  • a differentiation inducer that acts on alveolar epithelial stem cells, comprising a VDR acting compound as an active ingredient.
  • the differentiation inducer differentiates alveolar epithelial stem cells into alveolar epithelial cells.
  • the differentiation inducer contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a pharmaceutical composition for inducing alveolar regeneration comprising a VDR acting compound as an active ingredient.
  • the pharmaceutical composition contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
  • the form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a pharmaceutical composition for treating lung tissue damage comprising a VDR acting compound as an active ingredient.
  • the pharmaceutical composition contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
  • the form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
  • the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline).
  • the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
  • a method for treating lung tissue damage comprising administering a pharmaceutical composition comprising a VDR acting compound to a patient having damage to lung tissue.
  • the pharmaceutical composition used for the treatment method is the pharmaceutical composition of [12] or the pharmaceutical composition of [13].
  • patients having damage to lung tissue include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, Examples include interstitial pneumonia, pulmonary tuberculosis sequelae, and pneumoconiosis.
  • the administration mode is not particularly limited, and various administration routes such as pulmonary administration, intravenous administration, and oral administration can be selected, and preferably pulmonary administration.
  • the dosage is selected according to the type of disease; patient symptom, body weight, age; mode of administration; type of VDR acting compound;
  • [15] to [18] and [15 '] to [18'] are provided.
  • [15] to [18] and [15 ′] to [18 ′] correspond to [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration] described later.
  • a VDR acting compound as an active ingredient.
  • the details of [15] to [18] and [15 ′] to [18 ′] are described later in [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration]. It is the same.
  • a lyophilized composition comprising a VDR acting compound, at least two types of amino acids, and a surfactant.
  • a lyophilized composition comprising a VDR acting compound, at least two amino acids selected from the following first group, second group and third group and a plurality of amino acids, and a surfactant.
  • ⁇ Group 1 phenylalanine, tryptophan
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • a lyophilized composition of a composition comprising a VDR acting compound, at least two types of amino acids, a surfactant, an alcohol, and water.
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • a method for producing a lyophilized composition comprising the steps of preparing a second liquid and lyophilizing the second liquid.
  • [17 ′] A step of preparing a first liquid by dissolving a VDR acting compound in a solvent containing alcohol, the first liquid, and a plurality of groups from the following first group, second group, and third group
  • a method for producing the composition ⁇ Group 1: phenylalanine, tryptophan ⁇
  • Group 2 isoleucine, leucine, valine
  • Group 3 alanine, glycine
  • a pharmaceutical composition for transpulmonary administration wherein the freeze-dried composition according to [15] or [16] is granulated by air flow.
  • [18 '] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [15'] or [16 '] is granulated by air flow.
  • [15], [16], [15 ′] or [16 ′] is filled into a transpulmonary administration device disclosed in, for example, US Pat. No. 7,735,485 B2 or US Pat. No. 4,822,709.
  • the following [19] or [19 ′] invention is provided.
  • a method for transpulmonary administration of a pharmaceutical composition for pulmonary administration comprising the step of preparing the pharmaceutical composition for pulmonary administration by granulating with air impact.
  • a differentiation inducer that acts on alveolar epithelial stem cells using a VDR acting compound as an active ingredient; a pharmaceutical composition that induces alveolar regeneration
  • a pharmaceutical composition for treating lung tissue damage a method for treating lung tissue damage; a lyophilized composition used in the manufacture of a pharmaceutical composition for treating lung tissue damage; suitable for transpulmonary administration by airflow;
  • a freeze-dried composition that is granulated into particles having different particle sizes; a method for producing the freeze-dried composition; a pharmaceutical composition for treating lung tissue damage suitable for transpulmonary administration; a pharmaceutical composition for treating lung tissue damage A method of transpulmonary administration of the product.
  • the lyophilized composition of the present invention is a lyophilized composition containing at least the following components.
  • ⁇ Drugs poorly soluble in water hereinafter sometimes referred to as “active ingredients”) -At least 2 types of amino acids-Surfactant
  • the lyophilized composition is a lyophilized composition of a composition (dispersion or lysate) containing at least the following components.
  • ⁇ Slightly soluble drug ⁇ At least two kinds of amino acids ⁇ Surfactant ⁇ Alcohol ⁇ Water And when the freeze-dried composition is made into particles by air flow, the proportion of particles having a particle size of 5 ⁇ m or less is 10%.
  • a freeze-dried composition containing a drug hardly soluble in water cannot be prepared, or it becomes a freeze-dried composition that is difficult to form particles even when subjected to airflow.
  • a lyophilized composition that particles into unsuitable large particles When at least two kinds of amino acids are not used, a freeze-dried composition containing a drug hardly soluble in water cannot be prepared, or it becomes a freeze-dried composition that is difficult to form particles even when subjected to airflow. Or a lyophilized composition that particles into unsuitable large particles.
  • the particle size is a geometric particle size or an aerodynamic particle size.
  • the above-mentioned ratio is the ratio (volume%) of particles having a particle diameter of 5 ⁇ m or less with respect to all the particles with respect to the geometric particle diameter.
  • the above ratio is the ratio (mass%) of the amount of drug contained in particles having a particle diameter of 5 ⁇ m or less with respect to the amount of drug contained in all particles (total amount of drug).
  • the lyophilized composition of the present invention is preferably a lyophilized composition containing at least the following components.
  • the lyophilized composition is preferably a lyophilized composition of a composition (dispersion or solution) containing at least the following components.
  • -A poorly water-soluble drug-At least two amino acids selected from the following group 1, group 2 and group 3 across multiple groups-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, Leucine, valine ⁇ Group 3: alanine, glycine ⁇ Surfactant ⁇ Alcohol ⁇ Water
  • a specific type of amino acid selected in the above combination hereinafter also referred to as “specific amino acid”.
  • the lyophilized composition of the present invention is a pre-preparation of a pharmaceutical composition for pulmonary administration (a lyophilized composition for the preparation of a pharmaceutical composition for pulmonary administration), and is granulated by air impact by inhalation of a patient, It produces particles with a geometric particle size of 5 ⁇ m or less.
  • the ratio of the particles having a geometric particle diameter of 5 ⁇ m or less to the total particles generated from the freeze-dried composition of the present invention is preferably 10% or more, more preferably 15% or more, even more preferably on a volume basis. Is 20% or more, more preferably 25% or more, and particularly preferably 30% or more.
  • the geometric particle diameter can be measured by, for example, a laser diffraction particle size distribution measuring apparatus.
  • the lyophilized composition of the present invention is a pre-preparation of a pharmaceutical composition for pulmonary administration (a lyophilized composition for the preparation of a pharmaceutical composition for pulmonary administration). Particles having a particle size of 5 ⁇ m or less are generated.
  • the ratio of the particles having an aerodynamic particle size of 5 ⁇ m or less to the total particles generated from the freeze-dried composition of the present invention is preferably 10% or more as the ratio (mass%) of the drug amount to the total drug amount. More preferably, it is 15% or more, more preferably 20% or more, still more preferably 25% or more, and particularly preferably 30% or more.
  • the aerodynamic particle size can be measured by, for example, Multi-stage Liquid Impinger, Andersen Cascade Impactor, Next Generation Pharmaceutical Impactor and the like.
  • the aerodynamic particle diameter of the particles needs to be a micro size of 5 ⁇ m or less. Since the aerodynamic particle size of the particles derived from the porous lyophilized composition is usually smaller than the geometric particle size, if the proportion of particles having a geometric particle size of 5 ⁇ m or less is within the above range, A sufficient proportion of particles of aerodynamic particle size suitable for pulmonary administration will be included.
  • the lyophilized composition of the present invention is preferably a porous solid from the viewpoint of being easily atomized by airflow.
  • the porous solid may be cracked, divided into several lumps, or may be partially broken as long as it can be granulated by an air stream.
  • -Drugs poorly soluble in water- “Slightly soluble in water” for a drug means a compound that is 10 mg or less soluble in 1 ml of water at 20 ° C., and the amount of the compound is preferably 5 mg or less, more preferably 1 mg or less, More preferably, it is 0.5 mg or less, Most preferably, it is 0.1 mg or less.
  • the poorly water-soluble drug include compounds having a differentiation-inducing action on alveolar epithelial stem cells.
  • the compound having a differentiation-inducing action on alveolar epithelial stem cells is not particularly limited as long as it has a pharmacological activity that induces differentiation of alveolar epithelial stem cells (preferably human alveolar epithelial stem cells).
  • alveolar epithelial stem cells when the compound is brought into contact with alveolar epithelial stem cells, at least one of type I alveolar epithelial cells and type II alveolar epithelial cells This can be confirmed by inducing the cells. Specifically, after culturing alveolar epithelial stem cells in a medium containing the compound, the cells were stained by immunostaining and observed with a fluorescence microscope, and AQP-5 (Aquaporin, a marker of type I alveolar epithelial cells) was observed.
  • AQP-5 Amporin, a marker of type I alveolar epithelial cells
  • SP-A Surfactant (apo) protein A, surfactant (apo) protein A
  • apo surfactant (apo) protein A
  • a marker of type II alveolar epithelial cells It can be confirmed by being.
  • Examples of compounds having differentiation-inducing action on alveolar epithelial stem cells include vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors. Include the following compounds.
  • Vitamin A derivatives for example, retinoids such as all-trans-retinoic acid (ATRA. Tretinoin), retinol, retinal, and synthetic retinoids (eg tamibarotene).
  • Provitamin A for example ⁇ -carotene, ⁇ -carotene Vitamin D 2 and its derivatives: eg ergosterol, ergocalciferol Vitamin D 3 and its derivatives: eg 7-dehydrocholesterol, previtamin D 3 , cholecalciferol, 25-hydroxycholecalciferol, 1,25-dihydroxycholecalciferol (calcidiol), 1,25-dihydroxyvitamin D 3 (calcitriol), calcitroic acid-vitamin D 4 and its derivatives: e.g., dihydro ergocalciferol Vita Emissions D 5 and derivatives thereof, vitamin D 6 and its derivatives, vitamin D 7 and its derivatives, vitamin D analogs: e.g., dihydro Taki sterol, calcipotriol, tacalcitol, direct inhibitor of paricalcitol-PI3K: for example, Wortmannin, LY294002, AS605240, ZSTK474,
  • examples of poorly water-soluble drugs that can be expected to have a pharmacological effect on the lung include steroids (beclomethasone, fluticasone, budesonide, etc.) and herbal turmeric components (curcumin).
  • the amount of the active ingredient contained in the lyophilized composition of the present invention is not particularly limited.
  • the amount of the active ingredient contained in the lyophilized composition for a single dose is, for example, 20 mg or less, preferably 10 mg or less, more preferably 5 mg or less, still more preferably 2 mg or less, and particularly preferably 1 mg or less.
  • the present invention uses at least two amino acids in combination.
  • glycine Gly
  • alanine Ala
  • valine Val
  • leucine Leu
  • isoleucine Ile
  • serine Ser
  • threonine Thr
  • aspartic acid Aspartic acid
  • glutamic acid Glu
  • Asn Asn
  • glutamine Gln
  • lysine Lys
  • arginine Arg
  • cysteine Cys
  • methione Met
  • phenylalanine Phe
  • tyrosine Tyr
  • tryptophan Trp
  • Histidine His
  • proline At least two types are selected from (Pro).
  • the 20 amino acids other than glycine may be D-type or L-type, or a mixture of D-type and L-type.
  • a preferable combination of at least two kinds of amino acids selected from the above amino acid group is that the lyophilized composition is easily formed into particles having a geometric particle diameter of 5 ⁇ m or less, and Phe or Trp / Ile, Leu, Val, At least one selected from Ala and Gly; at least one selected from Ile, Leu and Val / at least one selected from Ala and Gly; Among these, at least one combination selected from Phe or Trp / Ile, Leu and Val is more preferable, and a combination of Phe / Ile, Phe / Leu, Phe / Val and Trp / Ile is particularly preferable.
  • the total content of amino acids in the lyophilized composition of the present invention is not particularly limited, but is, for example, 0.001% by mass or more and less than 100% by mass, preferably 0.01% by mass or more and less than 100% by mass, more Preferably they are 0.1 mass% or more and less than 100 mass%, More preferably, they are 1 mass% or more and less than 100 mass%, Especially preferably, they are 10 mass% or more and less than 100% mass.
  • the present invention it is preferable to use at least two kinds of amino acids selected from the following first group, second group and third group as a plurality of groups as at least two kinds of amino acids.
  • the combination of a plurality of groups is a combination of the first group and the second group, a combination of the first group and the third group, a combination of the second group and the third group, and a combination of the first group, the second group and the third group. Any combination may be used.
  • First group phenylalanine (Phe), tryptophan (Trp)
  • Second group isoleucine (Ile), leucine (Leu), valine (Val)
  • Group 3 alanine (Ala), glycine (Gly)
  • the amino acids other than glycine may be D-type or L-type, or a mixture of D-type and L-type.
  • At least one of the two types is preferably an amino acid selected from the first group in that the freeze-dried composition is easily granulated into particles having a geometric particle size of 5 ⁇ m or less, that is, Phe or Trp Is preferred.
  • a preferred combination of at least two types of amino acids is at least selected from Phe or Trp / Ile, Leu, Val, Ala and Gly in that the lyophilized composition is easily granulated into particles having a geometric particle size of 5 ⁇ m or less.
  • the total content of the specific amino acids in the lyophilized composition of the present invention is not particularly limited, but is, for example, 0.001% by mass or more and less than 100% by mass, preferably 0.01% by mass or more and less than 100% by mass, More preferably, they are 0.1 mass% or more and less than 100 mass%, More preferably, they are 1 mass% or more and less than 100 mass%, Especially preferably, they are 10 mass% or more and less than 100% mass.
  • the freeze-dried composition of the present invention may contain an amino acid other than the specific amino acid when at least two kinds of amino acids are the specific amino acid.
  • a drug that is sparingly soluble in water is used by dissolving it in a solvent containing alcohol. Since alcohol volatilizes by lyophilization, it is usually not included in lyophilized compositions. Examples of the alcohol include ethanol, methanol, isopropyl alcohol, ethylene glycol, and the like, and preferably ethanol. These may be used alone or in combination of two or more.
  • any anionic surfactant, cationic surfactant, or nonionic surfactant can be used as long as it is a surfactant that is usually used in pharmaceuticals.
  • nonionic surfactants such as polyoxyethylene sorbitan fatty acid ester (for example, Tween surfactant) and sorbitan trioleate are exemplified.
  • surfactant examples include sucrose fatty acid ester, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, polyoxyethylene hydrogenated castor oil 140, polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60.
  • the composition (dispersion or solution) subjected to freeze-drying is a liquid containing water as a main solvent. Since all or most of the water is lost by the lyophilization process, the water is not included in the lyophilized composition, or a small amount, if any, is included.
  • the lyophilized composition of the present invention may contain other components in addition to the above components as long as the final preparation (pharmaceutical composition for pulmonary administration) does not affect the human body.
  • Other components include stabilization of active ingredients in the composition (dispersion or solution) to be lyophilized, stabilization of active ingredients after lyophilization, prevention of adsorption of active ingredients to containers, etc.
  • Pharmaceutical additives commonly used in the art for the purpose excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, etc. May be used.
  • the lyophilized composition of the present invention may contain a carrier other than amino acids.
  • carriers other than amino acids monosaccharides such as glucose; disaccharides such as sucrose, maltose, lactose and trehalose; oligosaccharides such as cyclodextrin; polysaccharides such as dextran 40 and pullulan; fatty acid sodium such as sodium caprate; Is mentioned. These may be used alone or in combination of two or more.
  • the amount of these carriers to be blended is not particularly limited as long as the lyophilized composition is granulated by an air stream. As a guide, it is blended so that the proportion of the lyophilized composition is 0.1% by mass or more and less than 100% by mass.
  • the method for producing a freeze-dried composition of the present invention includes freeze-drying a composition (dispersion or solution) containing at least the following components. ⁇ Slightly soluble drugs ⁇ At least two amino acids ⁇ Surfactant ⁇ Alcohol ⁇ Water
  • the method for producing a freeze-dried composition of the present invention preferably includes freeze-drying a composition (dispersion or solution) containing at least the following components.
  • a composition (dispersion or solution) containing at least the following components.
  • Group 3 Alanine, glycine ⁇ Surfactant ⁇ Alcohol ⁇ Water
  • the method for producing a lyophilized composition of the present invention preferably includes the following steps (A) to (C). According to the production method including the steps (A) to (C), it is possible to prepare a liquid (dispersion or solution) containing water as a main solvent in which the active ingredient is well dispersed or dissolved. A porous lyophilized composition can be produced that is granulated into a size suitable for transpulmonary administration by airflow.
  • the step (B) is preferably the following step (B ′).
  • a second liquid by mixing the following:-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, leucine, valine-Group 3: alanine, glycine
  • Step (A) is a step of preparing a first liquid in which a poorly water-soluble drug (active ingredient) is dissolved in a solvent containing alcohol.
  • the solvent containing alcohol may be a solvent of alcohol alone or an aqueous alcohol solution.
  • the alcohol include ethanol, methanol, isopropyl alcohol, ethylene glycol, and the like, and preferably ethanol.
  • the alcohol concentration of the solvent containing alcohol is not particularly limited as long as the active ingredient can be dissolved. However, since it is desirable that the second liquid to be subjected to freeze-drying has a low alcohol concentration in terms of easy freeze-drying, the alcohol concentration of the solvent containing alcohol may be adjusted from the above viewpoint.
  • the solvent containing alcohol may contain an organic solvent other than alcohol as a solubilizer, as long as the final preparation (pharmaceutical composition for pulmonary administration) does not affect the human body.
  • organic solvent include acetone.
  • step (B) preferably step (B ′)
  • the first liquid in which the active ingredient is dissolved, at least two kinds of amino acids (preferably specific amino acids), a surfactant, and water are mixed.
  • This is a step of preparing a second liquid (dispersion or solution). Since the first liquid usually has an alcohol concentration that is so high that it is difficult to freeze-dry, it is necessary to add water to lower the alcohol concentration to adjust the alcohol concentration to be easily freeze-dried.
  • Step (B) is a step of adjusting the alcohol concentration by adding water while favorably dispersing or dissolving the active ingredient with at least two types of amino acids (preferably specific amino acids) and a surfactant.
  • the order of mixing the first liquid, the amino acid and the surfactant is not limited, but the dispersibility or solubility of the active ingredient in the second liquid is better. And the aspect which mixes a 1st liquid and surfactant and mixes an amino acid after that is preferable. Specifically, a surfactant is added to the first liquid to prepare a solution containing the active ingredient and the surfactant, and this solution is mixed with an amino acid-containing solution in which an amino acid is dissolved or dispersed in water. Embodiments are preferred.
  • the concentration of the active ingredient in the second liquid is, for example, 0.001 mg / mL to 1000 mg / mL, preferably 0.001 mg / mL to 100 mg / mL, more preferably It is 0.001 mg / mL to 10 mg / mL, more preferably 0.001 mg / mL to 1 mg / mL, and particularly preferably 0.001 mg / mL to 0.1 mg / mL.
  • the total amount of amino acids relative to the amount of the active ingredient is, for example, 50 parts by weight to 80000 parts by weight, preferably 100 parts by weight to 40000 parts by weight, when the amount of the active ingredient is 100 parts by weight.
  • the amount is preferably 200 parts by weight to 20000 parts by weight, more preferably 400 parts by weight to 10,000 parts by weight, and particularly preferably 800 parts by weight to 8000 parts by weight.
  • the total concentration of amino acids in the second liquid is not particularly limited. For example, what is necessary is just to adjust according to the quantity and kind of active ingredient, for example, 0.001 mg / mL or more, 0.01 mg / mL or more, and 0.1 mg / mL or more.
  • the total concentration of the amino acids in the second liquid is preferably 10 mg / mL or less, more preferably 8 mg / mL or less, and even more preferably 4 mg in that the lyophilized composition is well porous and easily atomized by airflow. / ML or less, particularly preferably 2 mg / mL or less.
  • the mass ratio of amino acids contained in the second liquid is, for example, in the range of 10:90 to 90:10, preferably 20:80. 80:20, more preferably 30:70 to 70:30, and still more preferably 40:60 to 60:40.
  • the concentration of the surfactant in the second liquid is, for example, 0.0001% by mass to 1% by mass, preferably 0.001% by mass to 0.5% by mass, More preferably, it is 0.005% by mass to 0.5% by mass, still more preferably 0.005% by mass to 0.3% by mass, and particularly preferably 0.005% by mass to 0.1% by mass.
  • the alcohol concentration of the second liquid is preferably 0.1% by mass to 20% by mass, more preferably 0.1% by mass to 10% by mass, still more preferably 0.5% by mass to 10% by mass, particularly preferably. 0.5 mass% to 5 mass%.
  • step (B) formulation additives (excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, etc.) are further mixed. May be. A carrier other than amino acids may be mixed.
  • Step (C) is a step of freeze-drying the second liquid obtained in step (B).
  • the method for freeze-drying the second liquid is not particularly limited, and may be, for example, a freeze-drying method that is conventionally applied to the production of a conventionally known freeze-dried drug (for example, an injection that is dissolved at the time of use).
  • the pharmaceutical composition for transpulmonary administration of the present invention is a powdery pharmaceutical composition obtained by granulating the lyophilized composition of the present invention into an air stream.
  • One aspect of the pharmaceutical composition for pulmonary administration of the present invention comprises a compound having an action of inducing differentiation of alveolar epithelial stem cells (preferably human alveolar epithelial stem cells) as an active ingredient.
  • the pharmaceutical composition for pulmonary administration of the present invention contains a compound having a differentiation-inducing action on alveolar epithelial stem cells, it can be used as a therapeutic agent for a disease exhibiting lung tissue damage.
  • the disease include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, interstitial pneumonia, pulmonary tuberculosis. Examples include sequelae and pneumoconiosis. Therefore, according to the pharmaceutical composition for transpulmonary administration of the present invention, a method for treating a disease exhibiting lung tissue damage is provided.
  • the therapeutic method is a method for treating pulmonary tissue damage, comprising pulmonary administration of the pharmaceutical composition for pulmonary administration of the present invention to a patient having damage to pulmonary tissue (for example, a patient having the above-mentioned diseases).
  • the dosage of the pharmaceutical composition for pulmonary administration of the present invention is selected according to the type of disease; patient symptom, body weight, age; type of compound contained as an active ingredient, and the like.
  • the pharmaceutical composition for transpulmonary administration of the present invention is prepared by pulverizing a freeze-dried composition for preparation with an air stream.
  • the flow rate of the air flow is 5 L / min to 300 L / min, more preferably 10 L / min to 200 L / min, still more preferably 10 L / min to 150 L / min, and particularly preferably 10 L / min to 100 L / min.
  • the flow rate of the airflow is 1 m / sec to 300 m / sec, more preferably 2 m / sec to 250 m / sec, still more preferably 5 m / sec to 200 m / sec, and particularly preferably 10 m / sec to 150 m / sec.
  • the pharmaceutical composition for pulmonary administration of the present invention can be formed into particles by the lyophilized composition for its preparation by a relatively low velocity and flow rate of air flow, and therefore, as an administration mode, for example, US Pat. No. 7,735,485 B2 or Patent
  • the transpulmonary administration system disclosed in Japanese Patent No. 4822709 can be employed. That is, if the freeze-dried composition is provided together with the device for transpulmonary administration disclosed in the above document, the user of the device attaches a vial of the freeze-dried composition to the device at the time of use, and the freeze-dried composition is inhaled by the patient's inspiration.
  • the product becomes particles suitable for transpulmonary administration and can be taken as it is by inhalation.
  • the lyophilized composition of the present invention is provided together with the above-mentioned device for pulmonary administration, for example, one embodiment of a method for pulmonary administration of the pharmaceutical composition for pulmonary administration is provided.
  • the pulmonary administration method when the patient inhales the pharmaceutical composition for pulmonary administration of the present invention, the patient applies airflow to the freeze-dried composition of the present invention to form particles that are suitable for pulmonary administration. The process of including.
  • the proportion of particles having a particle size of 5 ⁇ m or less in the total particles is preferably 10% or more, more preferably 15% or more, still more preferably 20% or more, More preferably, it is 25% or more, and particularly preferably 30% or more.
  • the particle diameter means a geometric particle diameter or an aerodynamic particle diameter.
  • the above-mentioned ratio is the ratio (volume%) of particles having a particle diameter of 5 ⁇ m or less with respect to all the particles with respect to the geometric particle diameter.
  • the above ratio is the ratio (mass%) of the amount of drug contained in particles having a particle diameter of 5 ⁇ m or less with respect to the amount of drug contained in all particles (total amount of drug).
  • the proportion (volume%) of particles having a geometric particle size of 5 ⁇ m or less in the total particles is preferably 10% or more, more preferably 15% or more, and further Preferably it is 20% or more, More preferably, it is 25% or more, Most preferably, it is 30% or more.
  • the ratio (mass%) of the drug amount contained in particles having an aerodynamic particle size of 5 ⁇ m or less with respect to the total drug amount is preferably 10% or more, more preferably. It is 15% or more, more preferably 20% or more, further preferably 25% or more, and particularly preferably 30% or more.
  • the particles in order for particles administered transpulmonarily to reach and deposit in the lung, the particles must have an aerodynamic particle size of 5 ⁇ m or less. Since the aerodynamic particle size of the particles derived from the porous lyophilized composition is usually smaller than the geometric particle size, if the proportion of particles having a geometric particle size of 5 ⁇ m or less is within the above range, A sufficient proportion of particles of aerodynamic particle size suitable for pulmonary administration will be included.
  • CD90 is a marker for alveolar epithelial stem cells
  • AQP-5 is a marker for type I alveolar epithelial cells
  • SP-A is a marker for type II alveolar epithelial cells.
  • Reference Example 1 Verification of differentiation-inducing action of ATRA on human alveolar epithelial stem cells
  • Human alveolar epithelial stem cells used in Reference Example 1 were derived from human lung alveolar epithelial stem cells (Fujino N, at. Al. Am. J) derived from patients with lung disease (biopsy in lung cancer or COPD or surgical tissue removed during surgery). Respir. Cell Mol. Biol., 46, 422-430 (2012)).
  • the human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
  • FBS bovine serum
  • MEM Amo Acids Solution
  • All-trans retinoic acid is exposed to human alveolar epithelial stem cells (1 ⁇ 10 1-4 cells / cm 2 ) in the culture at a concentration of 0 ⁇ M, 0.01 ⁇ M, 0.1 ⁇ M, 1 ⁇ M, or 10 ⁇ M.
  • the culture was stationary for 5 days in a 37 ° C./5% CO 2 environment.
  • the cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution.
  • PBS Phosphate Buffered Saline
  • goat anti-human CD90 antibody SantaSCruz
  • goat anti-human AQP-5 antibody Santa ⁇ Cruz
  • goat anti-human SP-A antibody Santa ⁇ Cruz
  • Anti-goat antibody manufactured by Santa Cruz
  • Example A1 Verification of differentiation-inducing effect of PI3K inhibitor on human alveolar epithelial stem cells
  • the human alveolar epithelial stem cells used in Example A1 are human alveolar epithelial stem cells (Fujino N, at. Al. Am. J) derived from patients with lung diseases (biopsy in lung cancer or COPD, or excised tissues at the time of surgery). Respir. Cell Mol. Biol., 46, 422-430 (2012)).
  • the human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
  • FBS bovine serum
  • MEM Amino Acids Solution
  • Human alveolar epithelial stem cells (1 ⁇ 10 1 to 4 cells / cm 2 ) in the culture medium are mixed with PI3K inhibitor Wortmannin at a concentration of 0 ⁇ M, 0.01 ⁇ M, 0.1 ⁇ M, 1 ⁇ M, or 10 ⁇ M. After exposure, the cells were statically cultured in a 37 ° C./5% CO 2 environment for 5 days.
  • the cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution.
  • PBS Phosphate Buffered Saline
  • goat anti-human CD90 antibody SantaSCruz
  • goat anti-human AQP-5 antibody Santa ⁇ Cruz
  • goat anti-human SP-A antibody Santa ⁇ Cruz
  • Anti-goat antibody manufactured by Santa Cruz
  • the PI3K inhibitor showed a strong differentiation-inducing action on alveolar epithelial stem cells at a lower concentration than ATRA.
  • the concentrations of 0.1 ⁇ M are compared, the PI3K inhibitor has a differentiation-inducing action on type I alveolar epithelial cells that is 3 times or more of ATRA, and has a differentiation-inducing action on type II alveolar epithelial cells that is twice or more. showed that.
  • Example A2 Verification of effect of PI3K inhibitor on COPD animal model
  • a 6-week-old ICR mouse male was administered pulmonary elastase (7.5 U, manufactured by Wako) suspended in 50 ⁇ L of PBS twice a week with a sonde for one week to prepare an emphysema model. Subsequently, wortmannin was transpulmonary administered twice a week for 3 weeks at a dose of 10 ⁇ g / kg each time with a sonde. Twenty-one days after the first administration of wortmannin, both lungs were removed, tissues were fixed with paraformaldehyde, sections were prepared with cryostat, and hematoxylin / eosin staining (HE staining) was performed.
  • pulmonary elastase 7.5 U, manufactured by Wako
  • wortmannin was transpulmonary administered twice a week for 3 weeks at a dose of 10 ⁇ g / kg each time with a sonde. Twenty-one days after the first administration of wortmannin, both lungs were removed, tissues were fixed with paraformalde
  • the method draws a number of random lines, places them on the alveolar image, finds the sum of the lengths of the sections separated by the alveolar walls of each random line, and divides this by the total number of broken lines. It is required by doing.
  • the number of divisions by the alveolar septum per visual field is set to 200 mm or more, this is measured in 30 visual fields (6 cross sections x 5 visual fields) per individual, the average is calculated, and the average alveolar wall distance of each mouse is obtained, The average of each group (6 individuals) was calculated, and statistical analysis was performed by t-test. The results are shown in Table 3 and FIG.
  • Human alveolar epithelial stem cells used in Examples B1 and B2 are patients with lung disease (extracted tissue at the time of biopsy or surgery in lung cancer or COPD) Derived from human alveolar epithelial stem cells (Fujino N, at. Al. Am. J. Respir. Cell Mol. Biol., 46, 422-430 (2012)).
  • the human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
  • Example B1 Verification of differentiation-inducing action of 1,25-dihydroxyvitamin D 3 on human alveolar epithelial stem cells (1)
  • 1,25-dihydroxyvitamin D 3 is exposed to human alveolar epithelial stem cells (1 ⁇ 10 1 to 4 cells / cm 2 ) in the culture at a concentration of 0 ⁇ M, 0.01 ⁇ M, 0.1 ⁇ M, 1 ⁇ M or 10 ⁇ M. Then, static culture was performed in a 37 ° C./5% CO 2 environment for 5 days.
  • the cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution.
  • PBS Phosphate Buffered Saline
  • goat anti-human CD90 antibody SantaSCruz
  • goat anti-human AQP-5 antibody Santa ⁇ Cruz
  • goat anti-human SP-A antibody Santa ⁇ Cruz
  • Anti-goat antibody manufactured by Santa Cruz
  • Example B1 is compared with the reference example 1, 1,25-dihydroxyvitamin D 3 is at a lower concentration than ATRA, it showed strong differentiation-inducing effect on alveolar epithelial stem cells.
  • 1,25-dihydroxyvitamin D 3 has a differentiation-inducing action on type I alveolar epithelial cells that is 7 times or more of ATRA, and more than 3 times type II alveolar epithelial cells. Showed differentiation-inducing action.
  • Example B2 Verification of differentiation-inducing action of 1,25-dihydroxyvitamin D 3 on human alveolar epithelial stem cells (2)
  • An experiment similar to Example B1 was conducted, except that the concentration of 1,25-dihydroxyvitamin D 3 was 10 ⁇ M and the culture days were 0 days, 1 day, 2 days, 4 days or 6 days. The results are shown in Table 5.
  • the cells were fixed with a 4% paraformaldehyde-PBS solution immediately after exposure to 1,25-dihydroxyvitamin D 3 and washed with PBS.
  • Example B3 1,25- Verification of effect on COPD animal model dihydroxyvitamin D 3] Experiments were conducted to confirm the effects of 1,25-dihydroxyvitamin D 3 using COPD model mice (elastase-induced emphysema mice).
  • a 6-week-old ICR mouse male was subjected to pulmonary administration of elastase (7.5 U, manufactured by Wako) suspended in 50 ⁇ L of PBS twice a week with a sonde for one week to prepare an emphysema model. Subsequently, 1,25-dihydroxyvitamin D 3 twice a week for three weeks, at a dose of each time 10 [mu] g / kg, and transpulmonary administration at sonde.
  • elastase 7.5 U, manufactured by Wako
  • the method draws a number of random lines, places them on the alveolar image, finds the sum of the lengths of the sections separated by the alveolar walls of each random line, and divides this by the total number of broken lines. It is required by doing.
  • the number of divisions by the alveolar septum per visual field is set to 200 mm or more, this is measured in 30 visual fields (6 cross sections x 5 visual fields) per individual, the average is calculated, and the average alveolar wall distance of each mouse is obtained, The average of each group (6 individuals) was calculated, and statistical analysis was performed by t-test. The results are shown in Table 6 and FIG.
  • the control group (without administration of 1,25-dihydroxyvitamin D 3 ) has a markedly expanded alveolar space and experimental pathological COPD lesions (significant emphysema) caused by elastase. It was confirmed.
  • 1,25-dihydroxyvitamin D 3 administration group the distance between the average alveolar wall were significantly lower. Low distances between the average alveolar wall shows that the alveoli is played, the alveoli is reproduced by the administration 1,25-dihydroxyvitamin D 3, the 1,25-dihydroxyvitamin D 3 It has been found to be effective in treating lung tissue damage.
  • [C] Lyophilized composition The amino acids used in Examples C1 to C4 are in the L form other than glycine.
  • Example C1 Preparation of lyophilized composition containing ATRA (1)
  • Preparation of lyophilized composition 4 mg of ATRA was dissolved in 1 mL of ethanol, and 20 ⁇ L of Tween 20 was added as a surfactant to prepare an ATRA solution.
  • amino acid aqueous solutions of Phe, Leu, Ile and Val were prepared. The amino acid concentration of the aqueous amino acid solution was 6 mg / mL for Phe, and 2.4 mg / mL for Leu, Ile, and Val.
  • an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 7, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
  • the final concentration of ethanol in each dispersion was 5% by weight.
  • Table 7 shows the final amounts of the active ingredients and amino acids of each dispersion and the final concentration of the surfactant.
  • the liquid volume of 1 vial dispersion is 0.5 mL.
  • 0.5 mL of this dispersion is put into a glass container (vial) having a body diameter of 18 mm and a volume of 2 mL, and is rapidly frozen under acetone-dry ice. Thereafter, a shelf-like freeze dryer (Freezone Triad, manufactured by LABCONCO) is used. And lyophilized to obtain a lyophilized composition.
  • a shelf-like freeze dryer Freezone Triad, manufactured by LABCONCO
  • the appearance of the lyophilized composition was evaluated according to the following evaluation criteria. The results are shown in Table 7.
  • 1 The freeze-dried composition is not completed or is extremely small.
  • Example C2 Preparation of lyophilized composition containing ATRA (2)
  • Example C2-1 and Comparative Example C2-1 1 mg of ATRA was dissolved in 0.25 mL of ethanol, and 2.5 ⁇ L of Tween 80 was added as a surfactant to prepare an ATRA solution.
  • amino acid aqueous solutions of Phe and Leu were prepared. The amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu. Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
  • Example C2-2 1 mg of ATRA was dissolved in 0.25 mL of ethanol, and 5 ⁇ L of Tween 80 was added as a surfactant to prepare an ATRA solution.
  • amino acid aqueous solutions of Phe and Leu were prepared.
  • the amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu.
  • an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
  • Example C2-3 and Comparative Example C2-3 5 mg of ATRA was dissolved in 1.25 mL of ethanol, and 25 ⁇ L of Tween 80 was added as a surfactant to prepare an ATRA solution.
  • amino acid aqueous solutions of Phe and Gly were prepared.
  • the amino acid concentration of the aqueous amino acid solution was 8 mg / mL for Phe and 8 mg / mL for Gly.
  • an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
  • the final concentration of ethanol in each dispersion was 5% by mass.
  • Table 8 shows the final amounts of the active ingredients and amino acids of each dispersion and the final concentration of the surfactant.
  • the liquid volume of 1 vial dispersion is 0.5 mL.
  • Example C3 Preparation of lyophilized composition containing ATRA (3)
  • 5 mg of ATRA was dissolved in 1.25 mL of ethanol, and 25 ⁇ L of Tween 80 was added as a surfactant to prepare an ATRA solution.
  • amino acid aqueous solutions of amino acids described in Table 9 were prepared. The amino acid concentration of the aqueous amino acid solution was 8 mg / mL.
  • an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 9, and the liquid volume was adjusted with water to prepare a dispersion in which ATRA was dispersed.
  • the final concentration of ethanol in each dispersion was 5% by weight.
  • Table 9 shows the final amounts of the active ingredients and amino acids and the final concentration of the surfactant in each dispersion.
  • the liquid volume of 1 vial dispersion is 0.5 mL.
  • Example C4 1,25- Preparation of lyophilized compositions that comprise the dihydroxyvitamin D 3] 0.1 mg of 1,25-dihydroxyvitamin D 3 was dissolved in 0.25 mL of ethanol, and 5 ⁇ L of Tween 20 was added as a surfactant and dissolved. Separately, amino acid aqueous solutions of Phe and Leu were prepared. The amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu.
  • an aqueous amino acid solution is added to the 1,25-dihydroxyvitamin D 3 solution so that the final amount of the amino acid is as shown in Table 10, and the liquid volume is adjusted with water, and 1,25-dihydroxyvitamin D 3 is added.
  • a solution (5 mL) in which was dissolved was prepared.
  • the final concentration of ethanol in each dispersion was 5% by weight.
  • Table 10 shows the final amounts of active ingredients and amino acids and the final concentration of the surfactant in each dispersion.
  • the liquid volume of 1 vial dispersion is 0.5 mL.
  • Example C1 From this dispersion, a freeze-dried composition was obtained in the same manner as in Example C1, and the appearance and particle formation were evaluated. The results are shown in Table 10.
  • Example C4 As is apparent from Table 10, the freeze-dried composition of Example C4 was granulated by an air flow, and became a powdered drug in which particles having a particle diameter of 5 ⁇ m or less accounted for 20% or more. This result shows that a drug suitable for pulmonary administration is prepared from the lyophilized composition of the present invention.

Abstract

A pharmaceutical composition which comprises a PI3 kinase inhibitor as an active ingredient. A pharmaceutical composition which comprises a compound that acts on a vitamin D receptor as an active ingredient. A freeze-dried composition of a composition which comprises a drug that has poor solubility to water, at least two kinds of amino acids, a surfactant, alcohol, and water.

Description

ＰＩ３キナーゼ阻害剤を含む医薬組成物、ビタミンＤ受容体に作用する化合物を含む医薬組成物、凍結乾燥組成物、凍結乾燥組成物の製造方法、及び経肺投与用医薬組成物PHARMACEUTICAL COMPOSITION CONTAINING PI3 KINASE INHIBITOR, PHARMACEUTICAL COMPOSITION CONTAINING COMPOUND HAVING VITAMIN D RECEPTOR, LYophilized Composition, Method for Producing Lyophilized Composition, and Pharmaceutical Composition for Transpulmonary Administration
　本発明は、ＰＩ３キナーゼ阻害剤を含む医薬組成物、ビタミンＤ受容体に作用する化合物を含む医薬組成物、経肺投与用医薬組成物を製造するための凍結乾燥組成物、該凍結乾燥組成物の製造方法、及び経肺投与用医薬組成物に関する。 The present invention relates to a pharmaceutical composition containing a PI3 kinase inhibitor, a pharmaceutical composition containing a compound acting on a vitamin D receptor, a lyophilized composition for producing a pharmaceutical composition for pulmonary administration, and the lyophilized composition And a pharmaceutical composition for pulmonary administration.
　肺組織に損傷を有する疾患である、慢性閉塞性肺疾患（chronic obstructive pulmonary disease、ＣＯＰＤ）、肺気腫、慢性気管支炎、急性肺損傷（acute respiratory distress syndrome、ＡＲＤＳ）、肺線維症、肺癌、間質性肺炎、肺結核後遺症、じん肺などは不可逆的な肺胞の破壊を呈する呼吸器疾患である。現在のところ肺胞の不可逆的破壊病変に対して根治的な治療薬は存在せず、症状に対する対処療法薬が使用されている。
　これら疾患に対する根治的な治療薬としては、肺胞上皮幹細胞を、肺胞を構成しているＩ型及びＩＩ型肺胞上皮細胞へと分化誘導し、肺胞を再生させる治療薬が有望と考えられている。 Chronic obstructive pulmonary disease (COPD), pulmonary emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, stroma Pneumonia, pulmonary tuberculosis sequelae, pneumoconiosis, etc. are respiratory diseases that exhibit irreversible alveolar destruction. At present, there is no curative treatment for irreversible destruction of alveoli, and symptomatic treatment is being used.
As a radical therapeutic agent for these diseases, a therapeutic agent that induces differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells constituting the alveoli and regenerates the alveoli is considered promising. It has been.
　肺胞の再生能を有する化合物として、ビタミンＡの誘導体であるオールトランスレチノイン酸が報告されている（例えば、Massaro G.D. et. al., Nature Med (1997) 3: 675-677.参照）。そこで、レチノイン酸を有効成分とする経口薬の臨床試験が試みられたが、経口投与では肺組織に到達する前に代謝されてしまう故に肺組織で有効な薬物濃度を維持しにくく、肺胞修復効果は観察されていない（例えば、Roth M.D. et. al., Chest (2006) 130: 1334-1345.参照）。 All-trans retinoic acid, which is a derivative of vitamin A, has been reported as a compound capable of regenerating alveoli (see, for example, MassaroDG.D. Et. Al., Nature Med (1997) 3: 675-677). Therefore, clinical trials of oral drugs containing retinoic acid as an active ingredient have been attempted. However, oral administration is metabolized before reaching the lung tissue, making it difficult to maintain an effective drug concentration in the lung tissue, and alveolar repair. No effect has been observed (see, for example, Roth MD et. Al., Chest (2006) 130: 1334-1345)).
　ところで、経肺投与用粉末薬剤の吸入システムとして、空気衝撃によって凍結乾燥組成物を粒子化する吸入システムがある（例えば、米国特許第７７３５４８５Ｂ２号明細書、及び特許第４８２２７０９号公報参照）。この吸入システムによれば、患者の吸気により凍結乾燥組成物が経肺投与に適した粒子となり、そのまま吸入して経肺投与が実施される。
　しかしながら、このシステムはタンパク質やペプチド等の熱に不安定で、かつ、水溶性薬物に適した凍結乾燥法を用いており、レチノイン酸等の水に難溶性の薬物の経肺製剤化には適していない。したがって、このシステムをレチノイン酸等の難溶性薬物に適用するためには新しい処方や製造方法を見出す必要がある。 By the way, as an inhalation system for powder medicine for pulmonary administration, there is an inhalation system that particles a freeze-dried composition by air impact (see, for example, US Pat. No. 7,735,485 B2 and US Pat. No. 4,822,709). According to this inhalation system, the freeze-dried composition becomes particles suitable for transpulmonary administration by inhalation of the patient, and inhalation is performed as it is to perform transpulmonary administration.
However, this system uses a lyophilization method suitable for water-soluble drugs, such as proteins and peptides, and is suitable for transpulmonary formulation of drugs that are poorly soluble in water such as retinoic acid. Not. Therefore, in order to apply this system to poorly soluble drugs such as retinoic acid, it is necessary to find a new formulation and manufacturing method.
　肺胞上皮幹細胞を肺胞上皮細胞へと分化誘導する化合物を見出し、その化合物を経肺投与用医薬組成物に製剤することができれば、肺組織の損傷を呈する疾患に対して肺胞の再生を作用点とする治療方法を提供することができる。
　また、経肺投与用医薬組成物の投与態様として、米国特許第７７３５４８５Ｂ２号明細書又は特許第４８２２７０９号公報に開示された経肺投与用粉末薬剤吸入システムを採ることができれば、上記疾患の患者に簡便に肺胞の再生療法を提供することができる。 If a compound that induces differentiation of alveolar epithelial stem cells into alveolar epithelial cells has been found and the compound can be formulated into a pharmaceutical composition for transpulmonary administration, regeneration of alveoli will be possible for diseases exhibiting lung tissue damage. It is possible to provide a treatment method having an action point.
In addition, as a mode of administration of a pharmaceutical composition for pulmonary administration, if the powder drug inhalation system for pulmonary administration disclosed in US Pat. No. 7,735,485B2 or US Pat. A regenerative therapy for alveoli can be provided simply.
　本発明は、上記状況のもとになされた。 The present invention was made under the above situation.
　上記状況のもと、肺胞上皮幹細胞を肺胞上皮細胞へと分化誘導する分化誘導剤が必要とされている。
　上記状況のもと、肺胞の再生を誘導して、肺組織の損傷を治療する医薬組成物が必要とされている。 Under the circumstances described above, there is a need for a differentiation inducer that induces differentiation of alveolar epithelial stem cells into alveolar epithelial cells.
Under the circumstances described above, there is a need for a pharmaceutical composition that induces alveolar regeneration to treat lung tissue damage.
　上記状況のもと、肺組織の損傷を治療する水に難溶性の薬物を含む、経肺投与に適した医薬組成物が必要とされている。
　上記状況のもと、水に難溶性の薬物を含む凍結乾燥組成物であり、気流によって経肺投与に適した粒子径の粒子に粒子化する凍結乾燥組成物が必要とされている。 Under the above circumstances, there is a need for a pharmaceutical composition suitable for transpulmonary administration, which contains a poorly water-soluble drug for treating lung tissue damage.
Under the circumstances described above, there is a need for a lyophilized composition containing a drug that is sparingly soluble in water and that is granulated into particles having a particle size suitable for transpulmonary administration by airflow.
　前記課題を達成するための具体的手段は以下のとおりである。
　（Ａ１）ＰＩ３キナーゼ阻害剤を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。
　（Ａ２）ＰＩ３キナーゼ阻害剤を有効成分として含む、肺胞再生を誘導する医薬組成物。
　（Ａ３）ＰＩ３キナーゼ阻害剤を有効成分として含む、肺組織の損傷治療用の医薬組成物。
　（Ａ４）肺胞上皮幹細胞に作用する分化誘導剤の製造における、ＰＩ３キナーゼ阻害剤の使用。
　（Ａ５）肺胞再生を誘導する医薬組成物の製造における、ＰＩ３キナーゼ阻害剤の使用。
　（Ａ６）肺組織の損傷治療用の医薬組成物の製造における、ＰＩ３キナーゼ阻害剤の使用。
　（Ａ７）肺組織の損傷の治療における使用のためのＰＩ３キナーゼ阻害剤。
　（Ａ８）肺組織に損傷を有する患者に、ＰＩ３キナーゼ阻害剤を有効成分として含む医薬組成物を投与することを含む、肺組織の損傷の治療方法。
　（Ａ９）前記ＰＩ３キナーゼ阻害剤が、ＰＩ３キナーゼを直接的に阻害する薬剤、ＰＩ３Ｋ－Ａｋｔ経路を阻害する薬剤、ＰＩ３Ｋ－ＢＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＩＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＴＡＰＰ１経路を阻害する薬剤、ＰＩ３Ｋ－ＤＡＰＰ経路を阻害する薬剤、ＰＩ３Ｋ－ＧＡＢ１／２経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｃ経路を阻害する薬剤、ＰＩ３Ｋ－Ｒｈｏ経路を阻害する薬剤、ＰＩ３Ｋ－ＡＲＦ経路を阻害する薬剤、ＰＩ３Ｋ－ＰＤＫ１経路を阻害する薬剤、ＰＩ３Ｋ－ＥＥＡ１経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｅｎｏｓｙｎ５経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｉｐ４経路を阻害する薬剤、ＰＩ３Ｋ－ＦＡＢ１経路を阻害する薬剤、ＰＩ３Ｋ－ｐ４０ｐｈｏｘ経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ３経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ１７経路を阻害する薬剤、及びＰＩ３Ｋ－ＳＧＫ３経路を阻害する薬剤から選ばれた少なくとも１種である、前記（Ａ１）～（Ａ８）。 Specific means for achieving the above object are as follows.
(A1) A differentiation inducer that acts on alveolar epithelial stem cells, comprising a PI3 kinase inhibitor as an active ingredient.
(A2) A pharmaceutical composition for inducing alveolar regeneration, comprising a PI3 kinase inhibitor as an active ingredient.
(A3) A pharmaceutical composition for treating lung tissue injury, comprising a PI3 kinase inhibitor as an active ingredient.
(A4) Use of a PI3 kinase inhibitor in the production of a differentiation inducer that acts on alveolar epithelial stem cells.
(A5) Use of a PI3 kinase inhibitor in the manufacture of a pharmaceutical composition that induces alveolar regeneration.
(A6) Use of a PI3 kinase inhibitor in the manufacture of a pharmaceutical composition for treating lung tissue damage.
(A7) PI3 kinase inhibitor for use in the treatment of lung tissue damage.
(A8) A method for treating lung tissue damage, comprising administering a pharmaceutical composition comprising a PI3 kinase inhibitor as an active ingredient to a patient having damage to lung tissue.
(A9) The PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- PI3, an agent that inhibits the p40phox pathway Agents that inhibit -SNX3 path is at least one selected from agents that inhibit drug, and PI3K-SGK3 pathway inhibits PI3K-SNX17 pathway, wherein (A1) ~ (A8).
　前記課題を達成するための具体的手段は以下のとおりである。
　（Ｂ１）ビタミンＤ受容体に作用する化合物を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。
　（Ｂ２）ビタミンＤ受容体に作用する化合物を有効成分として含む、肺胞再生を誘導する医薬組成物。
　（Ｂ３）ビタミンＤ受容体に作用する化合物を有効成分として含む、肺組織の損傷治療用の医薬組成物。
　（Ｂ４）肺胞上皮幹細胞に作用する分化誘導剤の製造における、ビタミンＤ受容体に作用する化合物の使用。
　（Ｂ５）肺胞再生を誘導する医薬組成物の製造における、ビタミンＤ受容体に作用する化合物の使用。
　（Ｂ６）肺組織の損傷治療用の医薬組成物の製造における、ビタミンＤ受容体に作用する化合物の使用。
　（Ｂ７）肺組織の損傷の治療における使用のためのビタミンＤ受容体に作用する化合物。
　（Ｂ８）肺組織に損傷を有する患者に、ビタミンＤ受容体に作用する化合物を有効成分として含む医薬組成物を投与することを含む、肺組織の損傷の治療方法。
　（Ｂ９）前記ビタミンＤ受容体に作用する化合物が、ビタミンＤ２及びその誘導体、ビタミンＤ３及びその誘導体、ビタミンＤ４及びその誘導体、ビタミンＤ５及びその誘導体、ビタミンＤ６及びその誘導体、ビタミンＤ７及びその誘導体から選ばれた少なくとも１種である、前記（Ｂ１）～（Ｂ８）。 Specific means for achieving the above object are as follows.
(B1) A differentiation inducer that acts on alveolar epithelial stem cells, comprising a compound that acts on a vitamin D receptor as an active ingredient.
(B2) A pharmaceutical composition for inducing alveolar regeneration, comprising a compound that acts on a vitamin D receptor as an active ingredient.
(B3) A pharmaceutical composition for treating lung tissue damage, comprising a compound that acts on a vitamin D receptor as an active ingredient.
(B4) Use of a compound that acts on a vitamin D receptor in the production of a differentiation inducer that acts on alveolar epithelial stem cells.
(B5) Use of a compound that acts on a vitamin D receptor in the manufacture of a pharmaceutical composition that induces alveolar regeneration.
(B6) Use of a compound that acts on a vitamin D receptor in the manufacture of a pharmaceutical composition for treating lung tissue damage.
(B7) Compounds that act on vitamin D receptors for use in the treatment of lung tissue damage.
(B8) A method for treating lung tissue damage, comprising administering to a patient having damage to lung tissue a pharmaceutical composition comprising a compound that acts on a vitamin D receptor as an active ingredient.
(B9) The compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and derivatives thereof, wherein (B1) ~ (B8).
　前記課題を達成するための具体的手段は以下のとおりである。
　（Ｃ１）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤とを含み、気流によって粒子化された際に、粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。
　（Ｃ１０１）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤とを含み、気流によって粒子化された際に、幾何学的粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。
　（Ｃ１０２）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤とを含み、気流によって粒子化された際に、空気力学的粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。 Specific means for achieving the above object are as follows.
(C1) Freeze-drying that contains a poorly water-soluble drug, at least two types of amino acids, and a surfactant, and the proportion of particles having a particle diameter of 5 μm or less becomes 10% or more when formed into particles by air flow Composition.
(C101) contains a poorly water-soluble drug, at least two types of amino acids, and a surfactant, and when formed into particles by an air stream, the proportion of particles having a geometric particle diameter of 5 μm or less is 10% or more. A lyophilized composition.
(C102) contains a poorly water-soluble drug, at least two kinds of amino acids, and a surfactant, and when formed into particles by an air stream, the proportion of particles having an aerodynamic particle diameter of 5 μm or less is 10% or more. A lyophilized composition.
　（Ｃ２）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物であり、気流によって粒子化された際に、粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。
　（Ｃ２０１）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物であり、気流によって粒子化された際に、幾何学的粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。
　（Ｃ２０２）水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物であり、気流によって粒子化された際に、空気力学的粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。 (C2) A lyophilized composition of a composition comprising a drug that is sparingly soluble in water, at least two types of amino acids, a surfactant, an alcohol, and water. A freeze-dried composition in which the proportion of particles having a diameter of 5 μm or less is 10% or more.
(C201) A freeze-dried composition of a composition comprising a poorly water-soluble drug, at least two types of amino acids, a surfactant, an alcohol, and water, A freeze-dried composition in which the proportion of particles having a particle size of 5 μm or less is 10% or more.
(C202) A freeze-dried composition of a composition comprising a poorly water-soluble drug, at least two types of amino acids, a surfactant, an alcohol, and water, A freeze-dried composition in which the proportion of particles having a mechanical particle size of 5 μm or less is 10% or more.
　（Ｃ３）水に難溶性の薬物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン
　（Ｃ４）水に難溶性の薬物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン (C3) A freeze-dried composition comprising a poorly water-soluble drug, at least two amino acids selected from the following first group, second group and third group and a plurality of groups, and a surfactant: .
-Group 1: Phenylalanine, tryptophan-Group 2: Isoleucine, leucine, valine-Group 3: Alanine, glycine (C4) Drugs poorly soluble in water, and the following groups 1, 2, 3 A freeze-dried composition of a composition comprising at least two amino acids selected from a plurality of groups, a surfactant, an alcohol, and water.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
　（Ｃ５）前記水に難溶性の薬物が、肺胞上皮幹細胞に対して分化誘導作用を有する化合物である前記（Ｃ１）～（Ｃ４）、（Ｃ１０１）、（Ｃ１０２）、（Ｃ２０１）、及び（Ｃ２０２）のいずれか１項に記載の凍結乾燥組成物。
　（Ｃ６）前記肺胞上皮幹細胞に対して分化誘導作用を有する化合物が、ビタミンＡ、ビタミンＡ誘導体、プロビタミンＡ、ビタミンＤ受容体に作用する化合物、及びＰＩ３キナーゼ阻害剤から選ばれた少なくとも１種である前記（Ｃ５）に記載の凍結乾燥組成物。
　（Ｃ６０１）前記ビタミンＤ受容体に作用する化合物が、ビタミンＤ２及びその誘導体、ビタミンＤ３及びその誘導体、ビタミンＤ４及びその誘導体、ビタミンＤ５及びその誘導体、ビタミンＤ６及びその誘導体、ビタミンＤ７及びその誘導体から選ばれた少なくとも１種である、前記（Ｃ６）に記載の凍結乾燥組成物。
　（Ｃ６０２）前記ＰＩ３キナーゼ阻害剤が、ＰＩ３キナーゼを直接的に阻害する薬剤、ＰＩ３Ｋ－Ａｋｔ経路を阻害する薬剤、ＰＩ３Ｋ－ＢＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＩＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＴＡＰＰ１経路を阻害する薬剤、ＰＩ３Ｋ－ＤＡＰＰ経路を阻害する薬剤、ＰＩ３Ｋ－ＧＡＢ１／２経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｃ経路を阻害する薬剤、ＰＩ３Ｋ－Ｒｈｏ経路を阻害する薬剤、ＰＩ３Ｋ－ＡＲＦ経路を阻害する薬剤、ＰＩ３Ｋ－ＰＤＫ１経路を阻害する薬剤、ＰＩ３Ｋ－ＥＥＡ１経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｅｎｏｓｙｎ５経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｉｐ４経路を阻害する薬剤、ＰＩ３Ｋ－ＦＡＢ１経路を阻害する薬剤、ＰＩ３Ｋ－ｐ４０ｐｈｏｘ経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ３経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ１７経路を阻害する薬剤、及びＰＩ３Ｋ－ＳＧＫ３経路を阻害する薬剤から選ばれた少なくとも１種である、前記（Ｃ６）に記載の凍結乾燥組成物。 (C5) The (C1) to (C4), (C101), (C102), (C201), and (C1), wherein the water-insoluble drug is a compound having a differentiation-inducing action on alveolar epithelial stem cells. The freeze-dried composition according to any one of C202).
(C6) The compound having a differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors The freeze-dried composition according to (C5), which is a seed.
(C601) The compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and its derivatives, freeze-dried composition according to the (C6).
(C602) the PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- an agent that inhibits the p40phox pathway, P Agents that inhibit 3K-SNX3 pathway is at least one selected from agents that inhibit drug, and PI3K-SGK3 pathway inhibits PI3K-SNX17 pathway, freeze-dried composition according to the (C6).
　（Ｃ７）水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程とを含む凍結乾燥組成物の製造方法。
　（Ｃ７０１）水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と界面活性剤とを混合したのち、さらに、少なくとも２種類のアミノ酸及び水を混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程とを含む凍結乾燥組成物の製造方法。
　（Ｃ８）水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程とを含む凍結乾燥組成物の製造方法。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン
　（Ｃ８０１）水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と界面活性剤とを混合したのち、さらに、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸並びに水を混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程とを含む凍結乾燥組成物の製造方法。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン (C7) A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, the first liquid, at least two kinds of amino acids, a surfactant, water, A method for producing a lyophilized composition comprising the steps of preparing a second liquid by mixing and lyophilizing the second liquid.
(C701) A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, and after mixing the first liquid and the surfactant, at least two kinds of A method for producing a lyophilized composition comprising a step of preparing a second liquid by mixing an amino acid and water, and a step of lyophilizing the second liquid.
(C8) A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol, the first liquid, and the following first group, second group, and third group Freezing comprising a step of preparing a second liquid by mixing at least two kinds of amino acids selected over a plurality of groups, a surfactant, and water, and a step of freeze-drying the second liquid A method for producing a dry composition.
-Group 1: Phenylalanine, tryptophan-Group 2: Isoleucine, leucine, valine-Group 3: Alanine, glycine (C801) A first liquid is prepared by dissolving a poorly water-soluble drug in a solvent containing alcohol. After mixing the first liquid and the surfactant, at least two kinds of amino acids selected from the first group, the second group, and the third group described below over a plurality of groups, and A method for producing a lyophilized composition comprising the steps of mixing water to prepare a second liquid, and lyophilizing the second liquid.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
　（Ｃ９）前記水に難溶性の薬物が、肺胞上皮幹細胞に対して分化誘導作用を有する化合物である前記（Ｃ７）、（Ｃ７０１）、（Ｃ８）及び（Ｃ８０１）のいずれか１項に記載の凍結乾燥組成物の製造方法。
　（Ｃ１０）前記肺胞上皮幹細胞に対して分化誘導作用を有する化合物が、ビタミンＡ、ビタミンＡ誘導体、プロビタミンＡ、ビタミンＤ受容体に作用する化合物、及びＰＩ３キナーゼ阻害剤から選ばれた少なくとも１種である前記（Ｃ９）に記載の凍結乾燥組成物の製造方法。
　（Ｃ１１１）前記ビタミンＤ受容体に作用する化合物が、ビタミンＤ２及びその誘導体、ビタミンＤ３及びその誘導体、ビタミンＤ４及びその誘導体、ビタミンＤ５及びその誘導体、ビタミンＤ６及びその誘導体、ビタミンＤ７及びその誘導体から選ばれた少なくとも１種である、前記（Ｃ１０）に記載の凍結乾燥組成物の製造方法。
　（Ｃ１１２）前記ＰＩ３キナーゼ阻害剤が、ＰＩ３キナーゼを直接的に阻害する薬剤、ＰＩ３Ｋ－Ａｋｔ経路を阻害する薬剤、ＰＩ３Ｋ－ＢＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＩＴＫ経路を阻害する薬剤、ＰＩ３Ｋ－ＴＡＰＰ１経路を阻害する薬剤、ＰＩ３Ｋ－ＤＡＰＰ経路を阻害する薬剤、ＰＩ３Ｋ－ＧＡＢ１／２経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｃ経路を阻害する薬剤、ＰＩ３Ｋ－Ｒｈｏ経路を阻害する薬剤、ＰＩ３Ｋ－ＡＲＦ経路を阻害する薬剤、ＰＩ３Ｋ－ＰＤＫ１経路を阻害する薬剤、ＰＩ３Ｋ－ＥＥＡ１経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｅｎｏｓｙｎ５経路を阻害する薬剤、ＰＩ３Ｋ－Ｒａｂｉｐ４経路を阻害する薬剤、ＰＩ３Ｋ－ＦＡＢ１経路を阻害する薬剤、ＰＩ３Ｋ－ｐ４０ｐｈｏｘ経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ３経路を阻害する薬剤、ＰＩ３Ｋ－ＳＮＸ１７経路を阻害する薬剤、及びＰＩ３Ｋ－ＳＧＫ３経路を阻害する薬剤から選ばれた少なくとも１種である、前記（Ｃ１０）に記載の凍結乾燥組成物の製造方法。 (C9) The drug according to any one of (C7), (C701), (C8) and (C801), wherein the poorly water-soluble drug is a compound having a differentiation-inducing action on alveolar epithelial stem cells. A method for producing a freeze-dried composition.
(C10) The compound having a differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors The method for producing a lyophilized composition according to (C9), which is a seed.
(C111) The compound acting on the vitamin D receptor is vitamin D 2 and derivatives thereof, vitamin D 3 and derivatives thereof, vitamin D 4 and derivatives thereof, vitamin D 5 and derivatives thereof, vitamin D 6 and derivatives thereof, vitamins is at least one member selected from D 7 and its derivatives, method for producing a freeze-dried composition according to the (C10).
(C112) The PI3 kinase inhibitor is an agent that directly inhibits PI3 kinase, an agent that inhibits the PI3K-Akt pathway, an agent that inhibits the PI3K-BTK pathway, an agent that inhibits the PI3K-ITK pathway, PI3K-TAPP1 Agents that inhibit the pathway, agents that inhibit the PI3K-DAPP pathway, agents that inhibit the PI3K-GAB1 / 2 pathway, agents that inhibit the PI3K-Rac pathway, agents that inhibit the PI3K-Rho pathway, and inhibitors the PI3K-ARF pathway Agents that inhibit the PI3K-PDK1 pathway, agents that inhibit the PI3K-EEA1 pathway, agents that inhibit the PI3K-Rabenosyn5 pathway, agents that inhibit the PI3K-Rabip4 pathway, agents that inhibit the PI3K-FAB1 pathway, PI3K- an agent that inhibits the p40phox pathway, P Production of the lyophilized composition according to (C10) above, which is at least one selected from an agent that inhibits the 3K-SNX3 pathway, an agent that inhibits the PI3K-SNX17 pathway, and an agent that inhibits the PI3K-SGK3 pathway Method.
　（Ｃ１１）前記（Ｃ１）～（Ｃ６）、（Ｃ１０１）、（Ｃ１０２）、（Ｃ２０１）、（Ｃ２０２）、（Ｃ６０１）、及び（Ｃ６０２）のいずれか１項に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 (C11) The lyophilized composition according to any one of (C1) to (C6), (C101), (C102), (C201), (C202), (C601), and (C602) A pharmaceutical composition for pulmonary administration, which is made into particles by the method.
　本発明によれば、肺胞上皮幹細胞に作用する分化誘導剤が提供される。
　また、本発明によれば、肺胞の再生を誘導する医薬組成物、及び肺組織の損傷治療用の医薬組成物が提供される。 According to the present invention, a differentiation inducer that acts on alveolar epithelial stem cells is provided.
The present invention also provides a pharmaceutical composition for inducing alveolar regeneration and a pharmaceutical composition for treating lung tissue damage.
　本発明によれば、水に難溶性の薬物を含む凍結乾燥組成物であり、気流によって経肺投与に適した粒子径の粒子に粒子化する凍結乾燥組成物が提供される。
　また、本発明によれば、肺胞の再生を誘導する医薬組成物の製造に使用される凍結乾燥組成物であり、気流によって経肺投与に適した粒子径の粒子に粒子化する凍結乾燥組成物が提供される。
　また、本発明によれば、水に難溶性の薬物を含む、経肺投与に適した医薬組成物が提供される。
　また、本発明によれば、経肺投与に適した、肺胞の再生を誘導する医薬組成物が提供される。 According to the present invention, there is provided a freeze-dried composition containing a drug that is sparingly soluble in water, and is formed into particles having a particle size suitable for transpulmonary administration by airflow.
Further, according to the present invention, a freeze-dried composition used for the production of a pharmaceutical composition for inducing regeneration of alveoli, the freeze-dried composition that is granulated into particles having a particle size suitable for transpulmonary administration by air flow Things are provided.
In addition, according to the present invention, there is provided a pharmaceutical composition suitable for pulmonary administration, which contains a drug hardly soluble in water.
The present invention also provides a pharmaceutical composition that induces alveolar regeneration suitable for transpulmonary administration.
実施例Ａ２のＣＯＰＤモデルマウスにおける、ワートマニン処置をした肺組織の切片のＨＥ染色像、及び平均肺胞壁間距離を示すグラフである。It is a graph which shows the HE dyeing | staining image of the section | slice of the lung tissue which carried out the wortmannin treatment in the COPD model mouse | mouth of Example A2, and the average distance between alveolar walls. 実施例Ｂ３のＣＯＰＤモデルマウスにおける、１，２５－ジヒドロキシビタミンＤ３処置をした肺組織の切片のＨＥ染色像、及び平均肺胞壁間距離を示すグラフである。2 is a graph showing HE-stained images of a section of lung tissue treated with 1,25-dihydroxyvitamin D 3 and the average distance between alveolar walls in the COPD model mouse of Example B3.
　以下に、本発明の実施の形態について説明する。これらの説明及び実施例は本発明を例示するものであり、本発明の範囲を制限するものではない。
　本明細書において「～」を用いて示された数値範囲は、「～」の前後に記載される数値をそれぞれ最小値及び最大値として含む範囲を示す。
　本明細書において「工程」との語は、独立した工程だけでなく、他の工程と明確に区別できない場合であってもその工程の所期の目的が達成されれば、本用語に含まれる。
　本明細書において「治療」とは、症状の改善であればよく、重症化の抑制及び、症状の軽減又は緩和もこの用語に包摂される。
　本明細書において「再生」とは、当技術分野で通常用いられる意味で使用される。 Embodiments of the present invention will be described below. These descriptions and examples are illustrative of the invention and are not intended to limit the scope of the invention.
In the present specification, a numerical range indicated by using “to” indicates a range including the numerical values described before and after “to” as the minimum value and the maximum value, respectively.
In this specification, the term “process” is not limited to an independent process, and is included in this term if the intended purpose of the process is achieved even when it cannot be clearly distinguished from other processes. .
In the present specification, the “treatment” may be any improvement of symptoms, and the term also includes suppression of severity and reduction or alleviation of symptoms.
In this specification, “regeneration” is used in the meaning normally used in this technical field.
　以下、本発明者によって、肺胞上皮幹細胞に対する分化誘導作用が初めて確認された化合物、及び該化合物を含む医薬組成物について詳述する。 Hereinafter, a compound for which differentiation-inducing action on alveolar epithelial stem cells has been confirmed by the present inventor for the first time and a pharmaceutical composition containing the compound will be described in detail.
［Ａ］ＰＩ３キナーゼ阻害剤を含む医薬組成物
　ＰＩ３キナーゼ（phosphatidylinositol-3 kinase、ＰＩ３Ｋ）は、イノシトールリン脂質のイノシトール環３位のヒドロキシル基（－ＯＨ基）のリン酸化を行う酵素である。イノシトールリン脂質は、真核生物の細胞膜の成分の一つであり、ＰＩ３Ｋをはじめとしたキナーゼ（リン酸化酵素）の触媒作用を受けてホスファチジルイノシトール３，４，５－三リン酸（ＰｔｄＩｎｓ（３，４，５）Ｐ３）となり、プロテインキナーゼＢ（ＰＫＢ）／Ａｋｔの活性化を起こすことで、細胞の生存、増殖、分化、細胞死の制御に関連している。 [A] Pharmaceutical composition containing PI3 kinase inhibitor PI3 kinase (phosphatidylinositol-3 kinase, PI3K) is an enzyme that phosphorylates the hydroxyl group (—OH group) at the 3-position of the inositol ring of inositol phospholipid. Inositol phospholipids are one of the components of eukaryotic cell membranes, and are catalyzed by kinases (phosphorylating enzymes) such as PI3K, so that phosphatidylinositol 3,4,5-triphosphate (PtdIns (3 , 4, 5) P3), and activation of protein kinase B (PKB) / Akt is associated with the control of cell survival, proliferation, differentiation, and cell death.
　ＰＩ３Ｋ阻害剤は、ＰＩ３キナーゼを直接的に阻害する薬剤、及びＰＩ３Ｋの下流シグナル経路を阻害する薬剤である。ＰＩ３Ｋの下流シグナル経路は、例えば、ＰＩ３Ｋ－Ａｋｔ経路、ＰＩ３Ｋ－ＢＴＫ経路、ＰＩ３Ｋ－ＩＴＫ経路、ＰＩ３Ｋ－ＴＡＰＰ１経路、ＰＩ３Ｋ－ＤＡＰＰ経路、ＰＩ３Ｋ－ＧＡＢ１／２経路、ＰＩ３Ｋ－Ｒａｃ経路、ＰＩ３Ｋ－Ｒｈｏ経路、ＰＩ３Ｋ－ＡＲＦ経路、ＰＩ３Ｋ－ＰＤＫ１経路、ＰＩ３Ｋ－ＥＥＡ１経路、ＰＩ３Ｋ－Ｒａｂｅｎｏｓｙｎ５経路、ＰＩ３Ｋ－Ｒａｂｉｐ４経路、ＰＩ３Ｋ－ＦＡＢ１経路、ＰＩ３Ｋ－ｐ４０ｐｈｏｘ経路、ＰＩ３Ｋ－ＳＮＸ３経路、ＰＩ３Ｋ－ＳＮＸ１７経路、ＰＩ３Ｋ－ＳＧＫ３経路である。これまで、ＰＩ３Ｋ阻害剤について、肺胞上皮幹細胞に対する分化誘導作用を有することは報告されていない。本発明者らは、ＰＩ３Ｋ阻害剤が、肺胞上皮幹細胞をＩ型及びＩＩ型肺胞上皮細胞へと分化誘導するという新しい知見を得た。 A PI3K inhibitor is an agent that directly inhibits PI3 kinase and an agent that inhibits a downstream signal pathway of PI3K. The downstream signal pathway of PI3K is, for example, PI3K-Akt pathway, PI3K-BTK pathway, PI3K-ITK pathway, PI3K-TAPP pathway, PI3K-DAPP pathway, PI3K-GAB1 / 2 pathway, PI3K-Rac pathway, PI3K-Rho pathway , PI3K-ARF pathway, PI3K-PDK1 pathway, PI3K-EEA1 pathway, PI3K-Rabenosyn5 pathway, PI3K-Rabip4 pathway, PI3K-FAB1 pathway, PI3K-p40phox pathway, PI3K-SNX3 pathway, PI3K-SNX17 pathway, PI3K-SGK pathway It is. Until now, it has not been reported that PI3K inhibitors have a differentiation-inducing action on alveolar epithelial stem cells. The present inventors have obtained a new finding that PI3K inhibitors induce differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells.
　ＰＩ３Ｋ阻害剤としては、例えば、以下の化合物が挙げられる。
　・ＰＩ３Ｋの直接的阻害剤：例えば、ワートマニン（Wortmannin）、ＬＹ２９４００２、ＡＳ６０５２４０、ＺＳＴＫ４７４、ＰＩＫ－７５　Ｈｙｄｒｏｃｈｌｏｒｉｄｅ、ＩＰＩ－１４５（ＩＮＫ１１９７）、ＧＤＣ－０９４１、ＣＡＬ－１０１（Ｉｄｅｌａｌｉｓｉｂ、ＧＳ－１１０１）、ＢＥＺ２３５（ＮＶＰ－ＢＥＺ２３５、Ｄａｃｔｏｌｉｓｉｂ）、ＢＫＭ１２０（ＮＶＰ－ＢＫＭ１２０、Ｂｕｐａｒｌｉｓｉｂ）、ＧＳＫ２６３６７７１、ＣＺＣ２４８３２、ＧＤＣ－００３２、ＶＳ－５５８４（ＳＢ２３４３）、ＴＧ１００７１３、ＢＹＬ７１９、ＣＵＤＣ－９０７、３－Ｍｅｔｈｙｌａｄｅｎｉｎｅ、ＹＭ２０１６３６、ＢＧＴ２２６（ＮＶＰ－ＢＧＴ２２６）、ＢＡＹ８０－６９４６（Ｃｏｐａｎｌｉｓｉｂ）、ＰＦ－０４６９１５０２、ＰＫＩ－４０２、ＣＨ５１３２７９９、ＧＤＣ－０９８０（ＲＧ７４２２）、ＮＵ７４４１ (ＫＵ－５７７８８）、ＡＳ－２５２４２４、ＡＳ－６０４８５０、ＣＡＹ１０５０５、ＧＳＫ２１２６４５８（ＧＳＫ４５８）、Ａ６６、ＰＦ－０５２１２３８４（ＰＫＩ－５８７）、Ｐａｌｏｍｉｄ５２９（Ｐ５２９）、ＰＩＫ－２９４、ＰＩＫ－２９３、ＳＡＲ２４５４０９（ＸＬ７６５）、ＰＩＫ－９３、ＡＺＤ６４８２、ＡＳ－６０５２４０、ＧＳＫ１０５９６１５、ＴＧ１００－１１５、ＩＣ－８７１１４、ＰＩＫ－７５、ＰＩＫ－９０、ＴＧＸ－２２１、ＸＬ１４７、ＰＩ－１０３、ＩＣ４８６０６８、及びこれらの誘導体
　・ＰＩ３Ｋ－Ａｋｔ経路阻害剤：例えば、ＭＫ－２２０６、ＧＳＫ６９０６９３、ＧＤＣ－００６８、Ａ－６７４５６３、ＣＣＴ１２８９３０、及びこれらの誘導体
　・ＰＩ３Ｋ－ＰＤＫ１経路阻害剤：例えば、ＢＸ－９１２、ＢＸ－７９５、ＯＳＵ－０３０１２、ＰＨＴ－４２７、及びこれらの誘導体 Examples of the PI3K inhibitor include the following compounds.
Direct inhibitors of PI3K: for example, Wortmannin, LY294002, AS605240, ZSTK474, PIK-75 Hydrochloride, IPI-145 (INK1197), GDC-0941, CAL-101 (Idelalisab, GS-1101), BEZ235 NVP-BEZ235, Dactolisib), BKM120 (NVP-BKM120, Buparlisb), GSK2636761, CZC24832, GDC-0032, VS-5684 (SB2343), TG100713, BYL719, CUDC-907, T ), BAY 80-6946 (Copanlist), PF-046 1502, PKI-402, CH5132799, GDC-0980 (RG7422), NU7441 (KU-57788), AS-252424, AS-604850, CAY10505, GSK212645 (GSK458), A66, PF-05212384 (PKI-587), Paloid-5875 P529), PIK-294, PIK-293, SAR245409 (XL765), PIK-93, AZD6482, AS-605240, GSK1059615, TG100-115, IC-87114, PIK-75, PIK-90, TGX-221, XL147, PI-103, IC486068, and derivatives thereof PI3K-Akt pathway inhibitors: For example, MK-2206, GSK690693, GDC-0068 , A-745563, CCT128930, and derivatives thereof PI3K-PDK1 pathway inhibitors: For example, BX-912, BX-795, OSU-03012, PHT-427, and derivatives thereof
　かかる知見に基づき、下記の［１］～［４］の発明が提供される。 Based on such knowledge, the following inventions [1] to [4] are provided.
［１］ＰＩ３Ｋ阻害剤を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。当該分化誘導剤は、肺胞上皮幹細胞を肺胞上皮細胞へと分化させる。
　当該分化誘導剤は、薬学的に許容し得る媒質中に、ＰＩ３Ｋ阻害剤の少なくとも１種を有効成分として含む。
　当該分化誘導剤の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [1] A differentiation inducer that acts on alveolar epithelial stem cells, comprising a PI3K inhibitor as an active ingredient. The differentiation inducer differentiates alveolar epithelial stem cells into alveolar epithelial cells.
The differentiation inducer contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
There are no particular limitations on the type of medium and formulation additive used in the preparation of the differentiation inducer. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［２］ＰＩ３Ｋ阻害剤を有効成分として含む、肺胞再生を誘導する医薬組成物。
　当該医薬組成物は、薬学的に許容し得る媒質中に、ＰＩ３Ｋ阻害剤の少なくとも１種を有効成分として含む。
　当該医薬組成物の形態は、特に制限されず、経口投与に適した、錠剤、顆粒剤、散剤、カプセル剤、懸濁剤、シロップ、乳剤、リモナーデ剤；注射用アンプル剤、注射用凍結乾燥粉末剤；経肺投与用乾燥粉末剤；等が挙げられる。
　当該医薬組成物の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [2] A pharmaceutical composition for inducing alveolar regeneration comprising a PI3K inhibitor as an active ingredient.
The pharmaceutical composition contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
The form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
There are no particular limitations on the type of medium and formulation additives used in the preparation of the pharmaceutical composition. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［３］ＰＩ３Ｋ阻害剤を有効成分として含む、肺組織の損傷治療用の医薬組成物。
　当該医薬組成物は、薬学的に許容し得る媒質中に、ＰＩ３Ｋ阻害剤の少なくとも１種を有効成分として含む。
　当該医薬組成物の形態は、特に制限されず、経口投与に適した、錠剤、顆粒剤、散剤、カプセル剤、懸濁剤、シロップ、乳剤、リモナーデ剤；注射用アンプル剤、注射用凍結乾燥粉末剤；経肺投与用乾燥粉末剤；等が挙げられる。
　当該医薬組成物の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [3] A pharmaceutical composition for treating lung tissue damage, comprising a PI3K inhibitor as an active ingredient.
The pharmaceutical composition contains at least one PI3K inhibitor as an active ingredient in a pharmaceutically acceptable medium.
The form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
There are no particular limitations on the type of medium and formulation additives used in the preparation of the pharmaceutical composition. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［４］肺組織に損傷を有する患者に、ＰＩ３Ｋ阻害剤を有効成分として含む医薬組成物を投与することを含む、肺組織の損傷の治療方法。当該治療方法に用いる医薬組成物は、前記［２］の医薬組成物、又は前記［３］の医薬組成物である。
　肺組織に損傷を有する患者としては、例えば、慢性閉塞性肺疾患（chronic obstructive pulmonary disease、ＣＯＰＤ）、肺気腫、慢性気管支炎、急性肺損傷（acute respiratory distress syndrome、ＡＲＤＳ）、肺線維症、肺癌、間質性肺炎、肺結核後遺症、じん肺などの患者が挙げられる。
　当該治療方法において、投与態様は、特に制限されず、経肺投与、静脈内投与、経口投与など種々の投与経路の選択が可能であるが、好ましくは経肺投与である。
　投与量は、疾患の種類；患者の症状、体重、年齢；投与態様；ＰＩ３Ｋ阻害剤の種類；等に応じて選択される。 [4] A method for treating lung tissue damage, comprising administering a pharmaceutical composition comprising a PI3K inhibitor as an active ingredient to a patient having damage to lung tissue. The pharmaceutical composition used for the treatment method is the pharmaceutical composition of [2] or the pharmaceutical composition of [3].
Examples of patients having damage to lung tissue include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, Examples include interstitial pneumonia, pulmonary tuberculosis sequelae, and pneumoconiosis.
In the treatment method, the administration mode is not particularly limited, and various administration routes such as pulmonary administration, intravenous administration, and oral administration can be selected, and preferably pulmonary administration.
The dose is selected depending on the type of disease; patient symptom, body weight, age; mode of administration; type of PI3K inhibitor;
　さらに、下記の［５］～［８］及び［５’］～［８’］の発明が提供される。［５］～［８］及び［５’］～［８’］は、後述する［凍結乾燥組成物］、［凍結乾燥組成物の製造方法］、及び［経肺投与用医薬組成物］に相当し、有効成分としてＰＩ３Ｋ阻害剤を含むものである。［５］～［８］及び［５’］～［８’］の詳細は、後述する［凍結乾燥組成物］、［凍結乾燥組成物の製造方法］、及び［経肺投与用医薬組成物］と同様である。 Furthermore, the following inventions [5] to [8] and [5 '] to [8'] are provided. [5] to [8] and [5 ′] to [8 ′] correspond to [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration] described later. And a PI3K inhibitor as an active ingredient. The details of [5] to [8] and [5 ′] to [8 ′] are described later in [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration]. It is the same.
［５］ＰＩ３Ｋ阻害剤と、少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。 [5] A lyophilized composition comprising a PI3K inhibitor, at least two types of amino acids, and a surfactant.
［５’］ＰＩ３Ｋ阻害剤と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [5 ′] A lyophilized composition comprising a PI3K inhibitor, at least two kinds of amino acids selected from the following first group, second group and third group and a plurality of groups, and a surfactant.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［６］ＰＩ３Ｋ阻害剤と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。 [6] A lyophilized composition comprising a PI3K inhibitor, at least two amino acids, a surfactant, an alcohol, and water.
［６’］ＰＩ３Ｋ阻害剤と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [6 ′] a PI3K inhibitor, at least two amino acids selected from the first group, the second group, and the third group described below across a plurality of groups, a surfactant, an alcohol, and water A lyophilized composition of the composition.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［７］ＰＩ３Ｋ阻害剤を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程と、を含む凍結乾燥組成物の製造方法。 [7] A step of preparing a first liquid by dissolving a PI3K inhibitor in a solvent containing alcohol, mixing the first liquid, at least two kinds of amino acids, a surfactant, and water. A method for producing a lyophilized composition comprising the steps of preparing a second liquid and lyophilizing the second liquid.
［７’］ＰＩ３Ｋ阻害剤を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程と、を含む凍結乾燥組成物の製造方法。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [7 ′] A step of preparing a first liquid by dissolving a PI3K inhibitor in a solvent containing alcohol; the first liquid; a plurality of groups from the following first group, second group, and third group: A step of preparing a second liquid by mixing at least two kinds of amino acids selected over the two, a surfactant and water, and a step of freeze-drying the second liquid. A method for producing the composition.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［８］前記［５］又は［６］に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 [8] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [5] or [6] is granulated by airflow.
［８’］前記［５’］又は［６’］に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 [8 '] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [5'] or [6 '] is granulated by an air stream.
　さらに、前記［５］、［６］、［５’］又は［６’］の発明を、例えば米国特許第７７３５４８５Ｂ２号明細書又は特許第４８２２７０９号公報に開示された経肺投与用デバイスに充填して提供すれば、下記の［９］又は［９’］の発明が提供される。 Further, the invention of [5], [6], [5 ′] or [6 ′] is filled into a transpulmonary administration device disclosed in, for example, US Pat. No. 7,735,485 B2 or Japanese Patent No. 4822709. The following [9] or [9 ′] invention is provided.
［９］患者が前記［８］に記載の経肺投与用医薬組成物を吸入する際に、前記［５］又は［６］に記載の凍結乾燥組成物を前記患者の吸気による空気衝撃で粒子化して前記経肺投与用医薬組成物を調製する工程を含む経肺投与用医薬組成物の経肺投与方法。 [9] When the patient inhales the pharmaceutical composition for pulmonary administration according to [8], the freeze-dried composition according to [5] or [6] A method for transpulmonary administration of a pharmaceutical composition for pulmonary administration, comprising the step of preparing the pharmaceutical composition for pulmonary administration.
［９’］患者が前記［８’］に記載の経肺投与用医薬組成物を吸入する際に、前記［５’］又は［６’］に記載の凍結乾燥組成物を前記患者の吸気による空気衝撃で粒子化して前記経肺投与用医薬組成物を調製する工程を含む経肺投与用医薬組成物の経肺投与方法。 [9 ′] When the patient inhales the pharmaceutical composition for transpulmonary administration described in [8 ′], the freeze-dried composition described in [5 ′] or [6 ′] is generated by inhalation of the patient. A method for transpulmonary administration of a pharmaceutical composition for pulmonary administration, comprising the step of preparing the pharmaceutical composition for pulmonary administration by granulating with air impact.
　前記［１］～［９］及び［５’］～［９’］によれば、ＰＩ３Ｋ阻害剤を有効成分として、肺胞上皮幹細胞に作用する分化誘導剤；肺胞再生を誘導する医薬組成物；肺組織の損傷治療用の医薬組成物；肺組織の損傷の治療方法；肺組織の損傷治療用の医薬組成物の製造に使用される凍結乾燥組成物であり、気流によって経肺投与に適した粒子径の粒子に粒子化する凍結乾燥組成物；前記凍結乾燥組成物の製造方法；経肺投与に適した、肺組織の損傷治療用の医薬組成物；肺組織の損傷治療用の医薬組成物の経肺投与方法；が提供される。 According to the above [1] to [9] and [5 ′] to [9 ′], a differentiation inducer that acts on alveolar epithelial stem cells, comprising a PI3K inhibitor as an active ingredient; a pharmaceutical composition that induces alveolar regeneration A pharmaceutical composition for treating lung tissue damage; a method for treating lung tissue damage; a lyophilized composition used in the manufacture of a pharmaceutical composition for treating lung tissue damage; suitable for transpulmonary administration by airflow; A freeze-dried composition that is granulated into particles having different particle sizes; a method for producing the freeze-dried composition; a pharmaceutical composition for treating lung tissue damage suitable for transpulmonary administration; a pharmaceutical composition for treating lung tissue damage A method of transpulmonary administration of the product.
　以下、本発明者によって、肺胞上皮幹細胞に対する分化誘導作用が初めて確認された化合物、及び該化合物を含む医薬組成物について詳述する。 Hereinafter, a compound for which differentiation-inducing action on alveolar epithelial stem cells has been confirmed by the present inventor for the first time and a pharmaceutical composition containing the compound will be described in detail.
［Ｂ］ビタミンＤ受容体に作用する化合物を含む医薬組成物
　１，２５－ジヒドロキシビタミンＤ３（カルシトリオール）は、慢性閉塞性肺疾患（chronic obstructive pulmonary disease、ＣＯＰＤ）の患者の肺組織において発現が低下することが報告されているが（Sundar IK. et. al., Biochem Biophys Res Commun. (2011) 406: 127-133.）、肺胞上皮幹細胞に対する分化誘導作用を有することは報告されていない。
　本発明者らは、１，２５－ジヒドロキシビタミンＤ３が、肺胞上皮幹細胞をＩ型及びＩＩ型肺胞上皮細胞へと分化誘導するという新しい知見を得た。
　１，２５－ジヒドロキシビタミンＤ３がビタミンＤ受容体（vitamin D receptor、ＶＤＲ）のリガンドであることから、ＶＤＲに作用する化合物（以下「ＶＤＲ作用化合物」と称する。）は、肺胞上皮幹細胞に対する分化誘導作用を有することが考えられた。
　ＶＤＲ作用化合物には、ＶＤＲに直接結合する化合物、例えば、ＶＤＲのリガンド、ＶＤＲのアゴニストが含まれる。また、ＶＤＲ作用化合物には、ＶＤＲに直接結合しない化合物、例えば、抗ＶＤＲリガンド抗体、ＶＤＲの下流のシグナル伝達を促進する化合物、ＶＤＲの下流のシグナル伝達経路を構成するタンパク質に作用する化合物、ＶＤＲアクチベーターとして機能する化合物、ＶＤＲアクチベーターインヒビターを阻害する化合物が含まれる。 [B] Pharmaceutical composition containing a compound that acts on vitamin D receptor 1,25-dihydroxyvitamin D 3 (calcitriol) is expressed in lung tissue of patients with chronic obstructive pulmonary disease (COPD) (Sundar IK. Et. Al., Biochem Biophys Res Commun. (2011) 406: 127-133.), But it has been reported to have differentiation-inducing action on alveolar epithelial stem cells. Absent.
The present inventors have obtained a new finding that 1,25-dihydroxyvitamin D 3 induces differentiation of alveolar epithelial stem cells into type I and type II alveolar epithelial cells.
Since 1,25-dihydroxyvitamin D 3 is a ligand of vitamin D receptor (VDR), a compound that acts on VDR (hereinafter referred to as “VDR acting compound”) is used for alveolar epithelial stem cells. It was considered to have a differentiation-inducing action.
VDR acting compounds include compounds that bind directly to VDR, eg, VDR ligands, VDR agonists. VDR acting compounds include compounds that do not directly bind to VDR, such as anti-VDR ligand antibodies, compounds that promote signal transduction downstream of VDR, compounds that act on proteins constituting signal transduction pathways downstream of VDR, VDR Compounds that act as activators, compounds that inhibit VDR activator inhibitors are included.
　ＶＤＲ作用化合物のうち、ＶＤＲのリガンド及びアゴニストとしては、例えばビタミンＤ及びその誘導体が挙げられ、具体的には以下の化合物が挙げられる。
　・ビタミンＤ２及びその誘導体：例えば、エルゴステロール、エルゴカルシフェロール
　・ビタミンＤ３及びその誘導体：例えば、７－デヒドロコレステロール、プレビタミンＤ３、コレカルシフェロール、２５－ヒドロキシコレカルシフェロール、１，２５－ジヒドロキシコレカルシフェロール（カルシジオール）、１，２５－ジヒドロキシビタミンＤ３（カルシトリオール）、カルシトロン酸
　・ビタミンＤ４及びその誘導体：例えば、ジヒドロエルゴカルシフェロール
　・ビタミンＤ５及びその誘導体
　・ビタミンＤ６及びその誘導体
　・ビタミンＤ７及びその誘導体
　・ビタミンＤアナログ：例えば、ジヒドロタキステロール、カルシポトリオール、タカルシトール、パリカルシトール Among VDR acting compounds, VDR ligands and agonists include, for example, vitamin D and derivatives thereof, and specifically include the following compounds.
Vitamin D 2 and its derivatives: eg ergosterol, ergocalciferol Vitamin D 3 and its derivatives: eg 7-dehydrocholesterol, previtamin D 3 , cholecalciferol, 25-hydroxycholecalciferol, 1,25 Dihydroxycholecalciferol (calcidiol), 1,25-dihydroxyvitamin D 3 (calcitriol), calcitronic acid Vitamin D 4 and its derivatives: eg dihydroergocalciferol Vitamin D 5 and its derivatives Vitamin D 6 And its derivatives Vitamin D 7 and its derivatives Vitamin D analogs: for example, dihydrotaxosterol, calcipotriol, tacalcitol, paricalcitol
　ＶＤＲ作用化合物のうち、ＶＤＲの下流のシグナル伝達経路を構成するタンパク質に作用する化合物としては、例えばＲＡＮＫＬ（Receptor activator of NF-kappa B ligand）阻害物質及びＮＦ－ｋＢ（Nuclear factor-kappa B）阻害物質が挙げられ、具体的には以下の化合物が挙げられる。
　・ＲＡＮＫＬ阻害物質：例えば、ＷＰ９ＱＹ、Ｏｓｔｅｏｐｒｏｔｅｇｅｒｉｎ（ＯＰＧ）、抗ＲＡＮＫＬ抗体（デノスマブ）、ＲＡＮＫＬｓｉＲＮＡ、ＲＡＮＫＬｓｈＲＮＡ
　・ＮＦ－ｋＢ阻害物質：例えば、（５Ｚ）－７－Ｏｘｏｚｅａｅｎｏｌ、（５Ｚ）－Ｚｅａｅｎｏｌ、Ａｎｄｒｏｇｒａｐｈｏｌｉｄｅ、Ａｕｒｏｔｈｉｏｍａｌａｔｅ、Ｃａｐｓａｉｃｉｎ、（Ｅ）－Ｃａｐｓａｉｃｉｎ、Ｅｖｏｄｉａｍｉｎｅ、抗ＮＦ－ｋＢ抗体、ＮＦ－ｋＢｓｉＲＮＡ、ＮＦ－ｋＢｓｈＲＮＡ Among VDR acting compounds, compounds acting on proteins constituting a signal transduction pathway downstream of VDR include, for example, RANKL (Receptor activator of NF-kappa B ligand) inhibitor and NF-kB (Nuclear factor-kappa B) inhibition. Examples of the material include the following compounds.
-RANKL inhibitor: For example, WP9QY, Osteoprotegerin (OPG), anti-RANKL antibody (denosumab), RANKL siRNA, RANKL shRNA
NF-kB inhibitors: for example, (5Z) -7-Oxoeaenol, (5Z) -Zeaenol, Andrographolide, Aurothiomalate, Capsaicin, (E) -Capsaicin, Evodiamine, anti-NF-kBsRNA, NF-kBsRNA, NF-kBsRNA
　ＶＤＲ作用化合物としては、１，２５－ジヒドロキシビタミンＤ３が好ましい。１，２５－ジヒドロキシビタミンＤ３は、ＡＴＲＡよりも低濃度で有意に肺胞上皮幹細胞に対する分化誘導作用を示す。 The VDR active compound, preferably 1,25-dihydroxyvitamin D 3. 1,25-dihydroxyvitamin D 3 shows the differentiation inducing effect on significantly alveolar epithelial stem cells at lower concentrations than ATRA.
　かかる知見に基づき、下記の［１１］～［１４］の発明が提供される。 Based on such knowledge, the following inventions [11] to [14] are provided.
［１１］ＶＤＲ作用化合物を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。当該分化誘導剤は、肺胞上皮幹細胞を肺胞上皮細胞へと分化させる。
　当該分化誘導剤は、薬学的に許容し得る媒質中に、ＶＤＲ作用化合物の少なくとも１種を有効成分として含む。
　当該分化誘導剤の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [11] A differentiation inducer that acts on alveolar epithelial stem cells, comprising a VDR acting compound as an active ingredient. The differentiation inducer differentiates alveolar epithelial stem cells into alveolar epithelial cells.
The differentiation inducer contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
There are no particular limitations on the type of medium and formulation additive used in the preparation of the differentiation inducer. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［１２］ＶＤＲ作用化合物を有効成分として含む、肺胞再生を誘導する医薬組成物。
　当該医薬組成物は、薬学的に許容し得る媒質中に、ＶＤＲ作用化合物の少なくとも１種を有効成分として含む。
　当該医薬組成物の形態は、特に制限されず、経口投与に適した、錠剤、顆粒剤、散剤、カプセル剤、懸濁剤、シロップ、乳剤、リモナーデ剤；注射用アンプル剤、注射用凍結乾燥粉末剤；経肺投与用乾燥粉末剤；等が挙げられる。
　当該医薬組成物の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [12] A pharmaceutical composition for inducing alveolar regeneration comprising a VDR acting compound as an active ingredient.
The pharmaceutical composition contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
The form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
There are no particular limitations on the type of medium and formulation additives used in the preparation of the pharmaceutical composition. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［１３］ＶＤＲ作用化合物を有効成分として含む、肺組織の損傷治療用の医薬組成物。
　当該医薬組成物は、薬学的に許容し得る媒質中に、ＶＤＲ作用化合物の少なくとも１種を有効成分として含む。
　当該医薬組成物の形態は、特に制限されず、経口投与に適した、錠剤、顆粒剤、散剤、カプセル剤、懸濁剤、シロップ、乳剤、リモナーデ剤；注射用アンプル剤、注射用凍結乾燥粉末剤；経肺投与用乾燥粉末剤；等が挙げられる。
　当該医薬組成物の調製に用いられる媒質及び製剤用添加物の種類は、特に制限されない。媒質としては、固体媒質（例えば、ゼラチン、乳糖）及び液体媒質（例えば、アルコール、水、生理食塩水）が挙げられる。製剤用添加物としては、賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤などが挙げられる。 [13] A pharmaceutical composition for treating lung tissue damage, comprising a VDR acting compound as an active ingredient.
The pharmaceutical composition contains at least one VDR acting compound as an active ingredient in a pharmaceutically acceptable medium.
The form of the pharmaceutical composition is not particularly limited, and is suitable for oral administration. Tablet, granule, powder, capsule, suspension, syrup, emulsion, limonade; ampoule for injection, freeze-dried powder for injection Agent; dry powder for transpulmonary administration; and the like.
There are no particular limitations on the type of medium and formulation additives used in the preparation of the pharmaceutical composition. Examples of the medium include a solid medium (for example, gelatin and lactose) and a liquid medium (for example, alcohol, water, and physiological saline). Examples of the additive for preparation include excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, and the like.
［１４］肺組織に損傷を有する患者に、ＶＤＲ作用化合物を含む医薬組成物を投与することを含む、肺組織の損傷の治療方法。当該治療方法に用いる医薬組成物は、前記［１２］の医薬組成物、又は前記［１３］の医薬組成物である。
　肺組織に損傷を有する患者としては、例えば、慢性閉塞性肺疾患（chronic obstructive pulmonary disease、ＣＯＰＤ）、肺気腫、慢性気管支炎、急性肺損傷（acute respiratory distress syndrome、ＡＲＤＳ）、肺線維症、肺癌、間質性肺炎、肺結核後遺症、じん肺などの患者が挙げられる。
　当該治療方法において、投与態様は、特に制限されず、経肺投与、静脈内投与、経口投与など種々の投与経路の選択が可能であるが、好ましくは経肺投与である。
　投与量は、疾患の種類；患者の症状、体重、年齢；投与態様；ＶＤＲ作用化合物の種類；等に応じて選択される。 [14] A method for treating lung tissue damage, comprising administering a pharmaceutical composition comprising a VDR acting compound to a patient having damage to lung tissue. The pharmaceutical composition used for the treatment method is the pharmaceutical composition of [12] or the pharmaceutical composition of [13].
Examples of patients having damage to lung tissue include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, Examples include interstitial pneumonia, pulmonary tuberculosis sequelae, and pneumoconiosis.
In the treatment method, the administration mode is not particularly limited, and various administration routes such as pulmonary administration, intravenous administration, and oral administration can be selected, and preferably pulmonary administration.
The dosage is selected according to the type of disease; patient symptom, body weight, age; mode of administration; type of VDR acting compound;
　さらに、下記の［１５］～［１８］及び［１５’］～［１８’］の発明が提供される。［１５］～［１８］及び［１５’］～［１８’］は、後述する［凍結乾燥組成物］、［凍結乾燥組成物の製造方法］、及び［経肺投与用医薬組成物］に相当し、有効成分としてＶＤＲ作用化合物を含むものである。［１５］～［１８］及び［１５’］～［１８’］の詳細は、後述する［凍結乾燥組成物］、［凍結乾燥組成物の製造方法］、及び［経肺投与用医薬組成物］と同様である。 Further, the following inventions [15] to [18] and [15 '] to [18'] are provided. [15] to [18] and [15 ′] to [18 ′] correspond to [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration] described later. And a VDR acting compound as an active ingredient. The details of [15] to [18] and [15 ′] to [18 ′] are described later in [Freeze-dried composition], [Production method of freeze-dried composition], and [Pharmaceutical composition for transpulmonary administration]. It is the same.
［１５］ＶＤＲ作用化合物と、少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。 [15] A lyophilized composition comprising a VDR acting compound, at least two types of amino acids, and a surfactant.
［１５’］ＶＤＲ作用化合物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [15 ′] A lyophilized composition comprising a VDR acting compound, at least two amino acids selected from the following first group, second group and third group and a plurality of amino acids, and a surfactant.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［１６］ＶＤＲ作用化合物と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。 [16] A lyophilized composition of a composition comprising a VDR acting compound, at least two types of amino acids, a surfactant, an alcohol, and water.
［１６’］ＶＤＲ作用化合物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [16 ′] a VDR acting compound, at least two amino acids selected from the following first group, second group, and third group, and a plurality of groups, a surfactant, an alcohol, and water A lyophilized composition of the composition.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［１７］ＶＤＲ作用化合物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程と、を含む凍結乾燥組成物の製造方法。 [17] A step of preparing a first liquid by dissolving a VDR acting compound in a solvent containing alcohol, mixing the first liquid, at least two kinds of amino acids, a surfactant, and water. A method for producing a lyophilized composition comprising the steps of preparing a second liquid and lyophilizing the second liquid.
［１７’］ＶＤＲ作用化合物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、前記第一の液体と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、前記第二の液体を凍結乾燥する工程と、を含む凍結乾燥組成物の製造方法。
　・第１群：フェニルアラニン、トリプトファン
　・第２群：イソロイシン、ロイシン、バリン
　・第３群：アラニン、グリシン [17 ′] A step of preparing a first liquid by dissolving a VDR acting compound in a solvent containing alcohol, the first liquid, and a plurality of groups from the following first group, second group, and third group A step of preparing a second liquid by mixing at least two kinds of amino acids selected over the two, a surfactant and water, and a step of freeze-drying the second liquid. A method for producing the composition.
・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
［１８］前記［１５］又は［１６］に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 [18] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [15] or [16] is granulated by air flow.
［１８’］前記［１５’］又は［１６’］に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 [18 '] A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to [15'] or [16 '] is granulated by air flow.
　さらに、前記［１５］、［１６］、［１５’］又は［１６’］の発明を、例えば米国特許第７７３５４８５Ｂ２号明細書又は特許第４８２２７０９号公報に開示された経肺投与用デバイスに充填して提供すれば、下記の［１９］又は［１９’］の発明が提供される。 Further, the invention of [15], [16], [15 ′] or [16 ′] is filled into a transpulmonary administration device disclosed in, for example, US Pat. No. 7,735,485 B2 or US Pat. No. 4,822,709. The following [19] or [19 ′] invention is provided.
［１９］患者が前記［１８］に記載の経肺投与用医薬組成物を吸入する際に、前記［１５］又は［１６］に記載の凍結乾燥組成物を前記患者の吸気による空気衝撃で粒子化して前記経肺投与用医薬組成物を調製する工程を含む経肺投与用医薬組成物の経肺投与方法。 [19] When the patient inhales the pharmaceutical composition for pulmonary administration according to [18], the freeze-dried composition according to [15] or [16] A method for transpulmonary administration of a pharmaceutical composition for pulmonary administration, comprising the step of preparing the pharmaceutical composition for pulmonary administration.
［１９’］患者が前記［１８’］に記載の経肺投与用医薬組成物を吸入する際に、前記［１５’］又は［１６’］に記載の凍結乾燥組成物を前記患者の吸気による空気衝撃で粒子化して前記経肺投与用医薬組成物を調製する工程を含む経肺投与用医薬組成物の経肺投与方法。 [19 ′] When the patient inhales the pharmaceutical composition for pulmonary administration described in [18 ′], the freeze-dried composition described in [15 ′] or [16 ′] is generated by inhalation of the patient. A method for transpulmonary administration of a pharmaceutical composition for pulmonary administration, comprising the step of preparing the pharmaceutical composition for pulmonary administration by granulating with air impact.
　前記［１１］～［１９］及び［１５’］～［１９’］によれば、ＶＤＲ作用化合物を有効成分として、肺胞上皮幹細胞に作用する分化誘導剤；肺胞再生を誘導する医薬組成物；肺組織の損傷治療用の医薬組成物；肺組織の損傷の治療方法；肺組織の損傷治療用の医薬組成物の製造に使用される凍結乾燥組成物であり、気流によって経肺投与に適した粒子径の粒子に粒子化する凍結乾燥組成物；前記凍結乾燥組成物の製造方法；経肺投与に適した、肺組織の損傷治療用の医薬組成物；肺組織の損傷治療用の医薬組成物の経肺投与方法；が提供される。 According to the above [11] to [19] and [15 ′] to [19 ′], a differentiation inducer that acts on alveolar epithelial stem cells using a VDR acting compound as an active ingredient; a pharmaceutical composition that induces alveolar regeneration A pharmaceutical composition for treating lung tissue damage; a method for treating lung tissue damage; a lyophilized composition used in the manufacture of a pharmaceutical composition for treating lung tissue damage; suitable for transpulmonary administration by airflow; A freeze-dried composition that is granulated into particles having different particle sizes; a method for producing the freeze-dried composition; a pharmaceutical composition for treating lung tissue damage suitable for transpulmonary administration; a pharmaceutical composition for treating lung tissue damage A method of transpulmonary administration of the product.
［Ｃ］凍結乾燥組成物、凍結乾燥組成物の製造方法、及び経肺投与用医薬組成物
［凍結乾燥組成物］
　本発明の凍結乾燥組成物は、少なくとも下記の成分を含む凍結乾燥組成物である。
　　・水に難溶性の薬物（以下「有効成分」という場合がある。）
　　・少なくとも２種類のアミノ酸
　　・界面活性剤
　当該凍結乾燥組成物は、少なくとも下記の成分を含む組成物（分散液又は溶解液）の凍結乾燥組成物である。
　　・水に難溶性の薬物
　　・少なくとも２種類のアミノ酸
　　・界面活性剤
　　・アルコール
　　・水
　そして、当該凍結乾燥組成物は、気流によって粒子化された際に、粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物である。
　少なくとも２種類のアミノ酸を使用しない場合には、水に難溶性の薬物を含む凍結乾燥組成物を作製できなかったり、気流を受けても粒子化しにくい凍結乾燥組成物に成ったり、経肺投与に適さない大きな粒子に粒子化する凍結乾燥組成物に成ったりする。 [C] Lyophilized composition, method for producing lyophilized composition, and pharmaceutical composition for pulmonary administration [lyophilized composition]
The lyophilized composition of the present invention is a lyophilized composition containing at least the following components.
・ Drugs poorly soluble in water (hereinafter sometimes referred to as “active ingredients”)
-At least 2 types of amino acids-Surfactant The lyophilized composition is a lyophilized composition of a composition (dispersion or lysate) containing at least the following components.
・ Slightly soluble drug ・ At least two kinds of amino acids ・ Surfactant ・ Alcohol ・ Water And when the freeze-dried composition is made into particles by air flow, the proportion of particles having a particle size of 5 μm or less is 10%. This is the lyophilized composition.
When at least two kinds of amino acids are not used, a freeze-dried composition containing a drug hardly soluble in water cannot be prepared, or it becomes a freeze-dried composition that is difficult to form particles even when subjected to airflow. Or a lyophilized composition that particles into unsuitable large particles.
　当該凍結乾燥組成物について、粒子径は、幾何学的粒子径または空気力学的粒子径である。
　上記の割合は、幾何学的粒子径については、全粒子に対する、粒子径５μｍ以下の粒子の割合（体積％）である。
　上記の割合は、空気力学的粒子径については、全粒子に含まれる薬物量（全薬物量）に対する、粒子径５μｍ以下の粒子に含まれる薬物量の割合（質量％）である。 For the lyophilized composition, the particle size is a geometric particle size or an aerodynamic particle size.
The above-mentioned ratio is the ratio (volume%) of particles having a particle diameter of 5 μm or less with respect to all the particles with respect to the geometric particle diameter.
The above ratio is the ratio (mass%) of the amount of drug contained in particles having a particle diameter of 5 μm or less with respect to the amount of drug contained in all particles (total amount of drug).
　本発明の凍結乾燥組成物は、好ましくは、少なくとも下記の成分を含む凍結乾燥組成物である。
　　・水に難溶性の薬物
　　・下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸
　　　　・第１群：フェニルアラニン、トリプトファン
　　　　・第２群：イソロイシン、ロイシン、バリン
　　　　・第３群：アラニン、グリシン
　　・界面活性剤
　当該凍結乾燥組成物は、好ましくは、少なくとも下記の成分を含む組成物（分散液又は溶解液）の凍結乾燥組成物である。
　　・水に難溶性の薬物
　　・下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸
　　　　・第１群：フェニルアラニン、トリプトファン
　　　　・第２群：イソロイシン、ロイシン、バリン
　　　　・第３群：アラニン、グリシン
　　・界面活性剤
　　・アルコール
　　・水
　少なくとも２種類のアミノ酸として、上記の組合せで選ばれた特定の種類のアミノ酸（以下「特定アミノ酸」ともいう。）を使用することによって、水に難溶性の薬物を含む凍結乾燥組成物が、気流を受けて経肺投与に適した粒子径により粒子化しやすい凍結乾燥組成物となる。 The lyophilized composition of the present invention is preferably a lyophilized composition containing at least the following components.
-A poorly water-soluble drug-At least two amino acids selected from the following group 1, group 2 and group 3 across multiple groups-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, Leucine, valine Group 3: Alanine, glycine Surfactant The lyophilized composition is preferably a lyophilized composition of a composition (dispersion or solution) containing at least the following components.
-A poorly water-soluble drug-At least two amino acids selected from the following group 1, group 2 and group 3 across multiple groups-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, Leucine, valine ・ Group 3: alanine, glycine ・ Surfactant ・ Alcohol ・ Water As at least two types of amino acids, a specific type of amino acid selected in the above combination (hereinafter also referred to as “specific amino acid”) is used. By doing so, a freeze-dried composition containing a drug that is sparingly soluble in water becomes a freeze-dried composition that easily undergoes particle formation with a particle size suitable for transpulmonary administration under airflow.
　本発明の凍結乾燥組成物は、経肺投与用医薬組成物の前調製物（経肺投与用医薬組成物の調製用の凍結乾燥組成物）であり、患者の吸気による空気衝撃で粒子化し、幾何学的粒子径が５μｍ以下の粒子を生じさせる。
　本発明の凍結乾燥組成物から生じた全粒子に占める幾何学的粒子径が５μｍ以下の粒子の割合は、体積基準で、１０％以上であることが好ましく、より好ましくは１５％以上、更に好ましくは２０％以上、更に好ましくは２５％以上、特に好ましくは３０％以上である。
　幾何学的粒子径は、例えば、レーザー回析式粒度分布測定装置によって測定し得る。 The lyophilized composition of the present invention is a pre-preparation of a pharmaceutical composition for pulmonary administration (a lyophilized composition for the preparation of a pharmaceutical composition for pulmonary administration), and is granulated by air impact by inhalation of a patient, It produces particles with a geometric particle size of 5 μm or less.
The ratio of the particles having a geometric particle diameter of 5 μm or less to the total particles generated from the freeze-dried composition of the present invention is preferably 10% or more, more preferably 15% or more, even more preferably on a volume basis. Is 20% or more, more preferably 25% or more, and particularly preferably 30% or more.
The geometric particle diameter can be measured by, for example, a laser diffraction particle size distribution measuring apparatus.
　本発明の凍結乾燥組成物は、経肺投与用医薬組成物の前調製物（経肺投与用医薬組成物の調製用の凍結乾燥組成物）であり、気流を受けて粒子化し、空気力学的粒子径が５μｍ以下の粒子を生じさせる。
　本発明の凍結乾燥組成物から生じた全粒子に占める空気力学的粒子径が５μｍ以下の粒子の割合は、全薬物量に対する薬物量の割合（質量％）として、１０％以上であることが好ましく、より好ましくは１５％以上、更に好ましくは２０％以上、更に好ましくは２５％以上、特に好ましくは３０％以上である。
　空気力学的粒子径は、例えば、Multi-stage Liquid Impinger、Andersen Cascade Impactor、Next Generation Pharmaceutical Impactor等によって測定し得る。 The lyophilized composition of the present invention is a pre-preparation of a pharmaceutical composition for pulmonary administration (a lyophilized composition for the preparation of a pharmaceutical composition for pulmonary administration). Particles having a particle size of 5 μm or less are generated.
The ratio of the particles having an aerodynamic particle size of 5 μm or less to the total particles generated from the freeze-dried composition of the present invention is preferably 10% or more as the ratio (mass%) of the drug amount to the total drug amount. More preferably, it is 15% or more, more preferably 20% or more, still more preferably 25% or more, and particularly preferably 30% or more.
The aerodynamic particle size can be measured by, for example, Multi-stage Liquid Impinger, Andersen Cascade Impactor, Next Generation Pharmaceutical Impactor and the like.
　一般に、経肺的に投与された粒子が肺に到達し沈着するには、その粒子の空気力学的粒子径が５μｍ以下のミクロサイズであることが必要と言われている。多孔質の凍結乾燥組成物に由来する粒子の空気力学的粒子径は通常、幾何学的粒子径よりも小さいので、幾何学的粒子径５μｍ以下の粒子の割合が上記の範囲であれば、経肺投与に適した空気力学的粒子径の粒子が十分な割合で含まれることになる。 Generally, in order for particles administered transpulmonarily to reach and deposit in the lung, it is said that the aerodynamic particle diameter of the particles needs to be a micro size of 5 μm or less. Since the aerodynamic particle size of the particles derived from the porous lyophilized composition is usually smaller than the geometric particle size, if the proportion of particles having a geometric particle size of 5 μm or less is within the above range, A sufficient proportion of particles of aerodynamic particle size suitable for pulmonary administration will be included.
　本発明の凍結乾燥組成物は、気流を受けて均一に粒子化しやすい点で、多孔質の固体であることが好ましい。多孔質の固体は、気流によって粒子化し得るものであれば、ひびが入ったり、数個の塊に分かれたり、一部が破損していてもよい。 The lyophilized composition of the present invention is preferably a porous solid from the viewpoint of being easily atomized by airflow. The porous solid may be cracked, divided into several lumps, or may be partially broken as long as it can be granulated by an air stream.
　以下、本発明の凍結乾燥組成物の調製に使用される各成分について詳述する。 Hereinafter, each component used for the preparation of the freeze-dried composition of the present invention will be described in detail.
－水に難溶性の薬物－
　薬物について「水に難溶性」とは、２０℃下、水１ｍｌに対して溶ける量が１０ｍｇ以下である化合物を意味し、当該化合物の前記量は、好ましくは５ｍｇ以下、より好ましくは１ｍｇ以下、更に好ましくは０．５ｍｇ以下、特に好ましくは０．１ｍｇ以下である。
　水に難溶性の薬物としては、例えば、肺胞上皮幹細胞に対して分化誘導作用を有する化合物が挙げられる。 -Drugs poorly soluble in water-
“Slightly soluble in water” for a drug means a compound that is 10 mg or less soluble in 1 ml of water at 20 ° C., and the amount of the compound is preferably 5 mg or less, more preferably 1 mg or less, More preferably, it is 0.5 mg or less, Most preferably, it is 0.1 mg or less.
Examples of the poorly water-soluble drug include compounds having a differentiation-inducing action on alveolar epithelial stem cells.
－肺胞上皮幹細胞に対して分化誘導作用を有する化合物－
　肺胞上皮幹細胞に対して分化誘導作用を有する化合物としては、肺胞上皮幹細胞（好ましくはヒト肺胞上皮幹細胞）の分化を誘導する薬理学的活性を有する化合物であれば特に制限されない。 -Compounds that have differentiation-inducing action on alveolar epithelial stem cells-
The compound having a differentiation-inducing action on alveolar epithelial stem cells is not particularly limited as long as it has a pharmacological activity that induces differentiation of alveolar epithelial stem cells (preferably human alveolar epithelial stem cells).
　ある化合物が、肺胞上皮幹細胞に対して分化誘導作用を有することは、当該化合物を肺胞上皮幹細胞に接触させた場合に、Ｉ型肺胞上皮細胞及びＩＩ型肺胞上皮細胞の少なくとも１種の細胞が誘導されることにより確認できる。具体的には、当該化合物を含む培地で肺胞上皮幹細胞を培養した後、免疫染色法によって細胞を染色して蛍光顕微鏡で観察し、Ｉ型肺胞上皮細胞のマーカーであるＡＱＰ－５（Aquaporin-5、アクアポリン５）、及びＩＩ型肺胞上皮細胞のマーカーであるＳＰ－Ａ（Surfactant (apo)protein A、サーファクタント（アポ）プロテインＡ）の少なくともいずれかを発現する細胞の数が増大していることによって確認できる。 The fact that a certain compound has a differentiation-inducing action on alveolar epithelial stem cells means that when the compound is brought into contact with alveolar epithelial stem cells, at least one of type I alveolar epithelial cells and type II alveolar epithelial cells This can be confirmed by inducing the cells. Specifically, after culturing alveolar epithelial stem cells in a medium containing the compound, the cells were stained by immunostaining and observed with a fluorescence microscope, and AQP-5 (Aquaporin, a marker of type I alveolar epithelial cells) was observed. -5, aquaporin 5), and the number of cells expressing at least one of SP-A (Surfactant (apo) protein A, surfactant (apo) protein A), a marker of type II alveolar epithelial cells, It can be confirmed by being.
　肺胞上皮幹細胞に対して分化誘導作用を有する化合物としては、例えば、ビタミンＡ、ビタミンＡ誘導体、プロビタミンＡ、ビタミンＤ受容体に作用する化合物、及びＰＩ３キナーゼ阻害剤が挙げられ、具体的には以下の化合物が挙げられる。
　・ビタミンＡ誘導体：例えば、オールトランスレチノイン酸（all-trans-retinoic acid、ＡＴＲＡ。トレチノイン）、レチノール、レチナール、及び合成レチノイド（例えば、タミバロテン）等のレチノイド
　・プロビタミンＡ：例えば、α－カロテン、β－カロテン
　・ビタミンＤ２及びその誘導体：例えば、エルゴステロール、エルゴカルシフェロール
　・ビタミンＤ３及びその誘導体：例えば、７－デヒドロコレステロール、プレビタミンＤ３、コレカルシフェロール、２５－ヒドロキシコレカルシフェロール、１，２５－ジヒドロキシコレカルシフェロール（カルシジオール）、１，２５－ジヒドロキシビタミンＤ３（カルシトリオール）、カルシトロン酸
　・ビタミンＤ４及びその誘導体：例えば、ジヒドロエルゴカルシフェロール
　・ビタミンＤ５及びその誘導体
　・ビタミンＤ６及びその誘導体
　・ビタミンＤ７及びその誘導体
　・ビタミンＤアナログ：例えば、ジヒドロタキステロール、カルシポトリオール、タカルシトール、パリカルシトール
　・ＰＩ３Ｋの直接的阻害剤：例えば、ワートマニン（Wortmannin）、ＬＹ２９４００２、ＡＳ６０５２４０、ＺＳＴＫ４７４、ＰＩＫ－７５　Ｈｙｄｒｏｃｈｌｏｒｉｄｅ、ＩＰＩ－１４５（ＩＮＫ１１９７）、ＧＤＣ－０９４１、ＣＡＬ－１０１（Ｉｄｅｌａｌｉｓｉｂ、ＧＳ－１１０１）、ＢＥＺ２３５（ＮＶＰ－ＢＥＺ２３５、Ｄａｃｔｏｌｉｓｉｂ）、ＢＫＭ１２０（ＮＶＰ－ＢＫＭ１２０、Ｂｕｐａｒｌｉｓｉｂ）、ＧＳＫ２６３６７７１、ＣＺＣ２４８３２、ＧＤＣ－００３２、ＶＳ－５５８４（ＳＢ２３４３）、ＴＧ１００７１３、ＢＹＬ７１９、ＣＵＤＣ－９０７、３－Ｍｅｔｈｙｌａｄｅｎｉｎｅ、ＹＭ２０１６３６、ＢＧＴ２２６（ＮＶＰ－ＢＧＴ２２６）、ＢＡＹ８０－６９４６（Ｃｏｐａｎｌｉｓｉｂ）、ＰＦ－０４６９１５０２、ＰＫＩ－４０２、ＣＨ５１３２７９９、ＧＤＣ－０９８０（ＲＧ７４２２）、ＮＵ７４４１ (ＫＵ－５７７８８）、ＡＳ－２５２４２４、ＡＳ－６０４８５０、ＣＡＹ１０５０５、ＧＳＫ２１２６４５８（ＧＳＫ４５８）、Ａ６６、ＰＦ－０５２１２３８４（ＰＫＩ－５８７）、Ｐａｌｏｍｉｄ５２９（Ｐ５２９）、ＰＩＫ－２９４、ＰＩＫ－２９３、ＳＡＲ２４５４０９（ＸＬ７６５）、ＰＩＫ－９３、ＡＺＤ６４８２、ＡＳ－６０５２４０、ＧＳＫ１０５９６１５、ＴＧ１００－１１５、ＩＣ－８７１１４、ＰＩＫ－７５、ＰＩＫ－９０、ＴＧＸ－２２１、ＸＬ１４７、ＰＩ－１０３、ＩＣ４８６０６８、及びこれらの誘導体
　・ＰＩ３Ｋ－Ａｋｔ経路阻害剤：例えば、ＭＫ－２２０６、ＧＳＫ６９０６９３、ＧＤＣ－００６８、Ａ－６７４５６３、ＣＣＴ１２８９３０、及びこれらの誘導体
　・ＰＩ３Ｋ－ＰＤＫ１経路阻害剤：例えば、ＢＸ－９１２、ＢＸ－７９５、ＯＳＵ－０３０１２、ＰＨＴ－４２７、及びこれらの誘導体 Examples of compounds having differentiation-inducing action on alveolar epithelial stem cells include vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors. Include the following compounds.
Vitamin A derivatives: for example, retinoids such as all-trans-retinoic acid (ATRA. Tretinoin), retinol, retinal, and synthetic retinoids (eg tamibarotene). Provitamin A: for example α-carotene, β-carotene Vitamin D 2 and its derivatives: eg ergosterol, ergocalciferol Vitamin D 3 and its derivatives: eg 7-dehydrocholesterol, previtamin D 3 , cholecalciferol, 25-hydroxycholecalciferol, 1,25-dihydroxycholecalciferol (calcidiol), 1,25-dihydroxyvitamin D 3 (calcitriol), calcitroic acid-vitamin D 4 and its derivatives: e.g., dihydro ergocalciferol Vita Emissions D 5 and derivatives thereof, vitamin D 6 and its derivatives, vitamin D 7 and its derivatives, vitamin D analogs: e.g., dihydro Taki sterol, calcipotriol, tacalcitol, direct inhibitor of paricalcitol-PI3K: for example, Wortmannin, LY294002, AS605240, ZSTK474, PIK-75 Hydrochloride, IPI-145 (INK1197), GDC-0941, CAL-101 (Idelalisib, GS-1101), BEZ235 (NVP-BEtVD235, NVP-BEZ235MN) -BKM120, Buparisib), GSK26367771, CZC24832, GDC-0032, VS-5484 (SB2343), TG10 713, BYL719, CUDC-907, 3-Methyladenine, YM201636, BGT226 (NVP-BGT226), BAY80-6946 (Copanlist), PF-04691502, PKI-402, CH5132799, GDC-0980 (RG74-8K) ), AS-252424, AS-604850, CAY10505, GSK2126458 (GSK458), A66, PF-05212384 (PKI-587), Palomid529 (P529), PIK-294, PIK-293, SAR245409 (XL765), PIK-93, AZD6482, AS-605240, GSK1059615, TG100-115, IC-87114, PIK- 5, PIK-90, TGX-221, XL147, PI-103, IC486068, and derivatives thereof PI3K-Akt pathway inhibitors: For example, MK-2206, GSK690693, GDC-0068, A-675633, CCT128930, and these Derivatives of PI3K-PDK1 pathway: for example, BX-912, BX-795, OSU-03012, PHT-427, and derivatives thereof
－その他の薬物－
　ほかに、肺への薬理学的効果が期待できる水に難溶性の薬物として、例えば、ステロイド（ベクロメタゾン、フルチカゾン、ブデソニド等）及び生薬ウコン成分（クルクミン）が挙げられる。 -Other drugs-
In addition, examples of poorly water-soluble drugs that can be expected to have a pharmacological effect on the lung include steroids (beclomethasone, fluticasone, budesonide, etc.) and herbal turmeric components (curcumin).
　本発明の凍結乾燥組成物中に含まれる有効成分の量は、特に制限されない。単回投与分の凍結乾燥組成物中に含まれる有効成分の量は、例えば２０ｍｇ以下であり、好ましくは１０ｍｇ以下、より好ましくは５ｍｇ以下、更に好ましくは２ｍｇ以下、特に好ましくは１ｍｇ以下である。 The amount of the active ingredient contained in the lyophilized composition of the present invention is not particularly limited. The amount of the active ingredient contained in the lyophilized composition for a single dose is, for example, 20 mg or less, preferably 10 mg or less, more preferably 5 mg or less, still more preferably 2 mg or less, and particularly preferably 1 mg or less.
－アミノ酸－
　本発明は、少なくとも２種類のアミノ酸を組み合わせて使用する。
　例えば、グリシン（Ｇｌｙ）、アラニン（Ａｌａ）、バリン（Ｖａｌ）、ロイシン（Ｌｅｕ）、イソロイシン（Ｉｌｅ）、セリン（Ｓｅｒ）、トレオニン（Ｔｈｒ）、アスパラギン酸（Ａｓｐ）、グルタミン酸（Ｇｌｕ）、アスパラギン（Ａｓｎ）、グルタミン（Ｇｌｎ）、リシン（Ｌｙｓ）、アルギニン（Ａｒｇ）、システイン（Ｃｙｓ）、メチオン（Ｍｅｔ）、フェニルアラニン（Ｐｈｅ）、チロシン（Ｔｙｒ）、トリプトファン（Ｔｒｐ）、ヒスチジン（Ｈｉｓ）、及びプロリン（Ｐｒｏ）から少なくとも２種類を選択する。
　グリシン以外の上記２０種類のアミノ酸は、Ｄ型でもＬ型でもよく、Ｄ型とＬ型の混合物でもよい。 -amino acid-
The present invention uses at least two amino acids in combination.
For example, glycine (Gly), alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), serine (Ser), threonine (Thr), aspartic acid (Asp), glutamic acid (Glu), asparagine ( Asn), glutamine (Gln), lysine (Lys), arginine (Arg), cysteine (Cys), methione (Met), phenylalanine (Phe), tyrosine (Tyr), tryptophan (Trp), histidine (His), and proline At least two types are selected from (Pro).
The 20 amino acids other than glycine may be D-type or L-type, or a mixture of D-type and L-type.
　上記のアミノ酸群から選択された少なくとも２種類のアミノ酸の好ましい組合せは、凍結乾燥組成物が幾何学的粒子径５μｍ以下の粒子に粒子化されやすい点で、Ｐｈｅ又はＴｒｐ／Ｉｌｅ、Ｌｅｕ、Ｖａｌ、Ａｌａ及びＧｌｙから選ばれた少なくとも１種；Ｉｌｅ、Ｌｅｕ及びＶａｌから選ばれた少なくとも１種／Ａｌａ及びＧｌｙから選ばれた少なくとも１種；である。
　中でも、Ｐｈｅ又はＴｒｐ／Ｉｌｅ、Ｌｅｕ及びＶａｌから選ばれた少なくとも１種の組合せがより好ましく、Ｐｈｅ／Ｉｌｅ、Ｐｈｅ／Ｌｅｕ、Ｐｈｅ／Ｖａｌ、Ｔｒｐ／Ｉｌｅの組合せが特に好ましい。 A preferable combination of at least two kinds of amino acids selected from the above amino acid group is that the lyophilized composition is easily formed into particles having a geometric particle diameter of 5 μm or less, and Phe or Trp / Ile, Leu, Val, At least one selected from Ala and Gly; at least one selected from Ile, Leu and Val / at least one selected from Ala and Gly;
Among these, at least one combination selected from Phe or Trp / Ile, Leu and Val is more preferable, and a combination of Phe / Ile, Phe / Leu, Phe / Val and Trp / Ile is particularly preferable.
　本発明の凍結乾燥組成物中の、アミノ酸の総含有量は、特に制限されないが、例えば０．００１質量％以上１００％質量未満であり、好ましくは０．０１質量％以上１００質量％未満、より好ましくは０．１質量％以上１００質量％未満、更に好ましくは１質量％以上１００質量％未満であり、特に好ましくは１０質量％以上１００％質量未満である。 The total content of amino acids in the lyophilized composition of the present invention is not particularly limited, but is, for example, 0.001% by mass or more and less than 100% by mass, preferably 0.01% by mass or more and less than 100% by mass, more Preferably they are 0.1 mass% or more and less than 100 mass%, More preferably, they are 1 mass% or more and less than 100 mass%, Especially preferably, they are 10 mass% or more and less than 100% mass.
－特定アミノ酸－
　本発明は、少なくとも２種類のアミノ酸として、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸を使用することが好ましい。複数群の組合せは、第１群と第２群の組合せ、第１群と第３群の組合せ、第２群と第３群の組合せ、及び、第１群と第２群と第３群の組合せのいずれでもよい。
　・第１群：フェニルアラニン（Ｐｈｅ）、トリプトファン（Ｔｒｐ）
　・第２群：イソロイシン（Ｉｌｅ）、ロイシン（Ｌｅｕ）、バリン（Ｖａｌ）
　・第３群：アラニン（Ａｌａ）、グリシン（Ｇｌｙ）
　グリシン以外の上記のアミノ酸は、Ｄ型でもＬ型でもよく、Ｄ型とＬ型の混合物でもよい。 -Specific amino acid-
In the present invention, it is preferable to use at least two kinds of amino acids selected from the following first group, second group and third group as a plurality of groups as at least two kinds of amino acids. The combination of a plurality of groups is a combination of the first group and the second group, a combination of the first group and the third group, a combination of the second group and the third group, and a combination of the first group, the second group and the third group. Any combination may be used.
First group: phenylalanine (Phe), tryptophan (Trp)
Second group: isoleucine (Ile), leucine (Leu), valine (Val)
Group 3: alanine (Ala), glycine (Gly)
The amino acids other than glycine may be D-type or L-type, or a mixture of D-type and L-type.
　少なくとも２種類のうち１種類は、凍結乾燥組成物が幾何学的粒子径５μｍ以下の粒子に粒子化されやすい点で、第１群から選ばれたアミノ酸であることが好ましく、即ち、Ｐｈｅ又はＴｒｐが好ましい。
　少なくとも２種類のアミノ酸の好ましい組合せは、凍結乾燥組成物が幾何学的粒子径５μｍ以下の粒子に粒子化されやすい点で、Ｐｈｅ又はＴｒｐ／Ｉｌｅ、Ｌｅｕ、Ｖａｌ、Ａｌａ及びＧｌｙから選ばれた少なくとも１種；Ｉｌｅ、Ｌｅｕ及びＶａｌから選ばれた少なくとも１種／Ａｌａ及びＧｌｙから選ばれた少なくとも１種；である。
　中でも、Ｐｈｅ又はＴｒｐ／Ｉｌｅ、Ｌｅｕ及びＶａｌから選ばれた少なくとも１種の組合せがより好ましく、Ｐｈｅ／Ｉｌｅ、Ｐｈｅ／Ｌｅｕ、Ｐｈｅ／Ｖａｌ、Ｔｒｐ／Ｉｌｅの組合せが特に好ましい。 At least one of the two types is preferably an amino acid selected from the first group in that the freeze-dried composition is easily granulated into particles having a geometric particle size of 5 μm or less, that is, Phe or Trp Is preferred.
A preferred combination of at least two types of amino acids is at least selected from Phe or Trp / Ile, Leu, Val, Ala and Gly in that the lyophilized composition is easily granulated into particles having a geometric particle size of 5 μm or less. At least one selected from Ile, Leu and Val / at least one selected from Ala and Gly;
Among these, at least one combination selected from Phe or Trp / Ile, Leu and Val is more preferable, and a combination of Phe / Ile, Phe / Leu, Phe / Val and Trp / Ile is particularly preferable.
　本発明の凍結乾燥組成物中の、特定アミノ酸の総含有量は、特に制限されないが、例えば０．００１質量％以上１００％質量未満であり、好ましくは０．０１質量％以上１００質量％未満、より好ましくは０．１質量％以上１００質量％未満、更に好ましくは１質量％以上１００質量％未満であり、特に好ましくは１０質量％以上１００％質量未満である。 The total content of the specific amino acids in the lyophilized composition of the present invention is not particularly limited, but is, for example, 0.001% by mass or more and less than 100% by mass, preferably 0.01% by mass or more and less than 100% by mass, More preferably, they are 0.1 mass% or more and less than 100 mass%, More preferably, they are 1 mass% or more and less than 100 mass%, Especially preferably, they are 10 mass% or more and less than 100% mass.
　本発明の凍結乾燥組成物は、少なくとも２種類のアミノ酸が特定アミノ酸である場合、特定アミノ酸以外のアミノ酸を含んでいてもよい。 The freeze-dried composition of the present invention may contain an amino acid other than the specific amino acid when at least two kinds of amino acids are the specific amino acid.
－アルコール－
　本発明の凍結乾燥組成物を製造する際、水に難溶性の薬物を、アルコールを含む溶媒に溶解させて使用する。
　アルコールは凍結乾燥処理により揮発するので、通常、凍結乾燥組成物には含まれない。
　アルコールとしては、例えば、エタノール、メタノール、イソプロピルアルコール、エチレングリコール等が挙げられ、好ましくはエタノールである。
　これらは１種を単独で又は２種以上を組み合わせて使用してよい。 -alcohol-
When producing the freeze-dried composition of the present invention, a drug that is sparingly soluble in water is used by dissolving it in a solvent containing alcohol.
Since alcohol volatilizes by lyophilization, it is usually not included in lyophilized compositions.
Examples of the alcohol include ethanol, methanol, isopropyl alcohol, ethylene glycol, and the like, and preferably ethanol.
These may be used alone or in combination of two or more.
－界面活性剤－
　界面活性剤としては、通常医薬品に使用される界面活性剤であれば、アニオン性界面活性剤、カチオン性界面活性剤、及び非イオン性界面活性剤のいずれであっても使用することができる。好ましくは、ポリオキシエチレンソルビタン脂肪酸エステル（例えば、Ｔｗｅｅｎ系界面活性剤）、ソルビタントリオレート等の非イオン性界面活性剤が例示される。 -Surfactant-
As the surfactant, any anionic surfactant, cationic surfactant, or nonionic surfactant can be used as long as it is a surfactant that is usually used in pharmaceuticals. Preferably, nonionic surfactants such as polyoxyethylene sorbitan fatty acid ester (for example, Tween surfactant) and sorbitan trioleate are exemplified.
　界面活性剤の具体例としては、ショ糖脂肪酸エステル、ステアリン酸ポリオキシル４０、ポリオキシエチレン硬化ヒマシ油１０、ポリオキシエチレン硬化ヒマシ油１４０、ポリオキシエチレン硬化ヒマシ油５０、ポリオキシエチレン硬化ヒマシ油６０、ポリオキシエチレン（４２）ポリアキシプロピレン（６７）グリコール、ポリオキシエチレン（５４）ポリアキシプロピレン（３９）グリコール、ポリオキシエチレン（１０５）ポリアキシプロピレン（５）グリコール、ポリオキシエチレン（１９６）ポリアキシプロピレン（６７）グリコール、ポリオキシエチレン（１６０）ポリアキシプロピレン（３０）グリコール、セスキオレイン酸ソルビタン、トリオレイン酸ソルビタン、モノステアリン酸ソルビタン、モノパルミチン酸ソルビタン、ポリソルベート２０、ポリソルベート４０、ポリソルベート６０、ポリソルベート６５、ポリソルベート８０、モノステアリン酸グリセリン、ラウリル硫酸ナトリウム、ラウロマクロゴール、モノラウリン酸ポリオキシエチレンソルビタン（Ｔｗｅｅｎ２０）、オレイン酸ポリオキシエチレンソルビタン（Ｔｗｅｅｎ８０）等が挙げられる。
　これらは１種を単独で又は２種以上を組み合わせて使用してよい。 Specific examples of the surfactant include sucrose fatty acid ester, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil 10, polyoxyethylene hydrogenated castor oil 140, polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60. , Polyoxyethylene (42) polyaxpropylene (67) glycol, polyoxyethylene (54) polyaxpropylene (39) glycol, polyoxyethylene (105) polyaxpropylene (5) glycol, polyoxyethylene (196) poly Axypropylene (67) glycol, polyoxyethylene (160) polyaxpropylene (30) glycol, sorbitan sesquioleate, sorbitan trioleate, sorbitan monostearate, sorbita monopalmitate , Polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, glyceryl monostearate, sodium lauryl sulfate, lauromacrogol, polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan oleate (Tween 80), etc. Can be mentioned.
These may be used alone or in combination of two or more.
－水－
　本発明の凍結乾燥組成物の製造において、凍結乾燥に供される組成物（分散液又は溶解液）は、水を主溶媒とする液体である。
　水は凍結乾燥処理により全部又は大部分が失われるので、凍結乾燥組成物には含まれないか、含まれるとしても微量である。 -water-
In the production of the freeze-dried composition of the present invention, the composition (dispersion or solution) subjected to freeze-drying is a liquid containing water as a main solvent.
Since all or most of the water is lost by the lyophilization process, the water is not included in the lyophilized composition, or a small amount, if any, is included.
－その他の成分－
　本発明の凍結乾燥組成物は、最終調製物（経肺投与用医薬組成物）が人体に影響を与えないことを限度として、上記成分に加えてその他の成分を含んでいてもよい。
　その他の成分としては、凍結乾燥に供される組成物（分散液又は溶解液）中での有効成分の安定化、凍結乾燥後の有効成分の安定化、容器への有効成分の吸着防止等を目的に、当技術分野で慣用される製剤用添加物（賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤など）を使用してよい。 -Other ingredients-
The lyophilized composition of the present invention may contain other components in addition to the above components as long as the final preparation (pharmaceutical composition for pulmonary administration) does not affect the human body.
Other components include stabilization of active ingredients in the composition (dispersion or solution) to be lyophilized, stabilization of active ingredients after lyophilization, prevention of adsorption of active ingredients to containers, etc. Pharmaceutical additives commonly used in the art for the purpose (excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, etc.) May be used.
　本発明の凍結乾燥組成物は、アミノ酸以外の担体を含んでいてもよい。
　アミノ酸以外の担体として、ブドウ糖等の単糖類；ショ糖、麦芽糖、乳糖、トレハロース等の二糖類；シクロデキストリン等のオリゴ糖類；デキストラン４０、プルラン等の多糖類；カプリン酸ナトリウム等の脂肪酸ナトリウム；などが挙げられる。これらは１種を単独で又は２種以上を組み合わせて使用してよい。
　これら担体の配合量は、凍結乾燥組成物が気流によって粒子化するものであれば、特に制限されない。目安として、凍結乾燥組成物に占める割合が０．１質量％以上１００質量％未満となるように配合する。 The lyophilized composition of the present invention may contain a carrier other than amino acids.
As carriers other than amino acids, monosaccharides such as glucose; disaccharides such as sucrose, maltose, lactose and trehalose; oligosaccharides such as cyclodextrin; polysaccharides such as dextran 40 and pullulan; fatty acid sodium such as sodium caprate; Is mentioned. These may be used alone or in combination of two or more.
The amount of these carriers to be blended is not particularly limited as long as the lyophilized composition is granulated by an air stream. As a guide, it is blended so that the proportion of the lyophilized composition is 0.1% by mass or more and less than 100% by mass.
［凍結乾燥組成物の製造方法］
　本発明の凍結乾燥組成物の製造方法は、少なくとも下記の成分を含む組成物（分散液又は溶解液）を凍結乾燥することを含む。
　　・水に難溶性の薬物
　　・少なくとも２種類のアミノ酸
　　・界面活性剤
　　・アルコール
　　・水 [Method for producing freeze-dried composition]
The method for producing a freeze-dried composition of the present invention includes freeze-drying a composition (dispersion or solution) containing at least the following components.
・ Slightly soluble drugs ・ At least two amino acids ・ Surfactant ・ Alcohol ・ Water
　本発明の凍結乾燥組成物の製造方法は、好ましくは、少なくとも下記の成分を含む組成物（分散液又は溶解液）を凍結乾燥することを含む。
　　・水に難溶性の薬物
　　・下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸
　　　　・第１群：フェニルアラニン、トリプトファン
　　　　・第２群：イソロイシン、ロイシン、バリン
　　　　・第３群：アラニン、グリシン
　　・界面活性剤
　　・アルコール
　　・水 The method for producing a freeze-dried composition of the present invention preferably includes freeze-drying a composition (dispersion or solution) containing at least the following components.
-A poorly water-soluble drug-At least two amino acids selected from the following group 1, group 2 and group 3 across multiple groups-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, Leucine, valine ・ Group 3: Alanine, glycine ・ Surfactant ・ Alcohol ・ Water
　本発明の凍結乾燥組成物の製造方法は、下記の工程（Ａ）～（Ｃ）を含むことが好ましい。工程（Ａ）～（Ｃ）を含む製造方法によれば、有効成分が良好に分散又は溶解した、水を主溶媒とする液体（分散液又は溶解液）を調製することができ、その結果、気流によって経肺投与に適した大きさに粒子化する多孔質の凍結乾燥組成物を製造できる。 The method for producing a lyophilized composition of the present invention preferably includes the following steps (A) to (C). According to the production method including the steps (A) to (C), it is possible to prepare a liquid (dispersion or solution) containing water as a main solvent in which the active ingredient is well dispersed or dissolved. A porous lyophilized composition can be produced that is granulated into a size suitable for transpulmonary administration by airflow.
　・工程（Ａ）：水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程
　・工程（Ｂ）：前記第一の液体と、少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程
　・工程（Ｃ）：前記第二の液体を凍結乾燥する工程 Step (A): a step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol. Step (B): the first liquid and at least two kinds of amino acids, Step of preparing a second liquid by mixing a surfactant and water-Step (C): Step of freeze-drying the second liquid
　工程（Ｂ）は、好ましくは以下の工程（Ｂ’）である。
　・工程（Ｂ’）：前記第一の液体と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程
　　　・第１群：フェニルアラニン、トリプトファン
　　　・第２群：イソロイシン、ロイシン、バリン
　　　・第３群：アラニン、グリシン The step (B) is preferably the following step (B ′).
Step (B ′): the first liquid, at least two kinds of amino acids selected from the following first group, second group, and third group across a plurality of groups, a surfactant, and water To prepare a second liquid by mixing the following:-Group 1: phenylalanine, tryptophan-Group 2: isoleucine, leucine, valine-Group 3: alanine, glycine
－工程（Ａ）－
　工程（Ａ）は、水に難溶性の薬物（有効成分）が、アルコールを含む溶媒に溶解した第一の液体を調製する工程である。 -Process (A)-
Step (A) is a step of preparing a first liquid in which a poorly water-soluble drug (active ingredient) is dissolved in a solvent containing alcohol.
　アルコールを含む溶媒は、アルコール単独の溶媒でもよく、アルコール水溶液でもよい。
　アルコールとしては、例えば、エタノール、メタノール、イソプロピルアルコール、エチレングリコール等が挙げられ、好ましくはエタノールである。
　アルコールを含む溶媒のアルコール濃度は、有効成分を溶解可能であれば特に制限されない。ただし、凍結乾燥に供する第二の液体は凍結乾燥しやすい点でアルコール濃度が低いことが望ましいので、上記観点でアルコールを含む溶媒のアルコール濃度を調整してよい。 The solvent containing alcohol may be a solvent of alcohol alone or an aqueous alcohol solution.
Examples of the alcohol include ethanol, methanol, isopropyl alcohol, ethylene glycol, and the like, and preferably ethanol.
The alcohol concentration of the solvent containing alcohol is not particularly limited as long as the active ingredient can be dissolved. However, since it is desirable that the second liquid to be subjected to freeze-drying has a low alcohol concentration in terms of easy freeze-drying, the alcohol concentration of the solvent containing alcohol may be adjusted from the above viewpoint.
　アルコールを含む溶媒は、最終調製物（経肺投与用医薬組成物）が人体に影響を与えないことを限度として、溶解補助剤としてアルコール以外の有機溶媒を含んでいてもよい。当該有機溶媒としては、例えば、アセトン等が例示される。 The solvent containing alcohol may contain an organic solvent other than alcohol as a solubilizer, as long as the final preparation (pharmaceutical composition for pulmonary administration) does not affect the human body. Examples of the organic solvent include acetone.
－工程（Ｂ）－
　工程（Ｂ）（好ましくは工程（Ｂ’））は、有効成分が溶解している第一の液体、少なくとも２種類のアミノ酸（好ましくは特定アミノ酸）、界面活性剤、及び水を混合して第二の液体（分散液又は溶解液）を調製する工程である。
　第一の液体は、通常、凍結乾燥しにくいほどにアルコール濃度が高いので、水を加えてアルコール濃度を低下させ、凍結乾燥しやすいアルコール濃度に調整する必要がある。工程（Ｂ）は、少なくとも２種類のアミノ酸（好ましくは特定アミノ酸）及び界面活性剤によって有効成分を良好に分散又は溶解させつつ、水を加えてアルコール濃度を調整する工程である。 -Process (B)-
In step (B) (preferably step (B ′)), the first liquid in which the active ingredient is dissolved, at least two kinds of amino acids (preferably specific amino acids), a surfactant, and water are mixed. This is a step of preparing a second liquid (dispersion or solution).
Since the first liquid usually has an alcohol concentration that is so high that it is difficult to freeze-dry, it is necessary to add water to lower the alcohol concentration to adjust the alcohol concentration to be easily freeze-dried. Step (B) is a step of adjusting the alcohol concentration by adding water while favorably dispersing or dissolving the active ingredient with at least two types of amino acids (preferably specific amino acids) and a surfactant.
　工程（Ｂ）において、第一の液体と、アミノ酸及び界面活性剤とを混合する順番は限定されるものではないが、第二の液体における有効成分の分散性又は溶解性がより良好である点で、第一の液体と界面活性剤を混合し、その後にアミノ酸を混合する態様が好ましい。具体的には、第一の液体に界面活性剤を添加して有効成分及び界面活性剤を含む溶液を調製し、この溶液と、アミノ酸を水に溶解又は分散させたアミノ酸含有液とを混合する態様が好ましい。 In the step (B), the order of mixing the first liquid, the amino acid and the surfactant is not limited, but the dispersibility or solubility of the active ingredient in the second liquid is better. And the aspect which mixes a 1st liquid and surfactant and mixes an amino acid after that is preferable. Specifically, a surfactant is added to the first liquid to prepare a solution containing the active ingredient and the surfactant, and this solution is mixed with an amino acid-containing solution in which an amino acid is dissolved or dispersed in water. Embodiments are preferred.
　第二の液体の有効成分の濃度（複数種使用する場合はその総濃度）は、例えば０．００１ｍｇ／ｍＬ～１０００ｍｇ／ｍＬであり、好ましくは０．００１ｍｇ／ｍＬ～１００ｍｇ／ｍＬ、より好ましくは０．００１ｍｇ／ｍＬ～１０ｍｇ／ｍＬ、更に好ましくは０．００１ｍｇ／ｍＬ～１ｍｇ／ｍＬ、特に好ましくは０．００１ｍｇ／ｍＬ～０．１ｍｇ／ｍＬである。 The concentration of the active ingredient in the second liquid (the total concentration when multiple types are used) is, for example, 0.001 mg / mL to 1000 mg / mL, preferably 0.001 mg / mL to 100 mg / mL, more preferably It is 0.001 mg / mL to 10 mg / mL, more preferably 0.001 mg / mL to 1 mg / mL, and particularly preferably 0.001 mg / mL to 0.1 mg / mL.
　第二の液体において、有効成分の量に対するアミノ酸の総量は、有効成分の量を１００質量部とした場合、例えば５０質量部～８００００質量部であり、好ましくは１００質量部～４００００質量部、より好ましくは２００質量部～２００００質量部、更に好ましくは４００質量部～１００００質量部、特に好ましくは８００質量部～８０００質量部である。 In the second liquid, the total amount of amino acids relative to the amount of the active ingredient is, for example, 50 parts by weight to 80000 parts by weight, preferably 100 parts by weight to 40000 parts by weight, when the amount of the active ingredient is 100 parts by weight. The amount is preferably 200 parts by weight to 20000 parts by weight, more preferably 400 parts by weight to 10,000 parts by weight, and particularly preferably 800 parts by weight to 8000 parts by weight.
　第二の液体のアミノ酸の総濃度は、特に限定されるものではない。例えば有効成分の量及び種類に応じて調整すればよく、例えば０．００１ｍｇ／ｍＬ以上であり、０．０１ｍｇ／ｍＬ以上であり、０．１ｍｇ／ｍＬ以上である。
　第二の液体のアミノ酸の総濃度は、凍結乾燥組成物の多孔質化が良好で気流によって粒子化されやすい点で、１０ｍｇ／ｍＬ以下が好ましく、より好ましくは８ｍｇ／ｍＬ以下、更に好ましくは４ｍｇ／ｍＬ以下、特に好ましくは２ｍｇ／ｍＬ以下である。 The total concentration of amino acids in the second liquid is not particularly limited. For example, what is necessary is just to adjust according to the quantity and kind of active ingredient, for example, 0.001 mg / mL or more, 0.01 mg / mL or more, and 0.1 mg / mL or more.
The total concentration of the amino acids in the second liquid is preferably 10 mg / mL or less, more preferably 8 mg / mL or less, and even more preferably 4 mg in that the lyophilized composition is well porous and easily atomized by airflow. / ML or less, particularly preferably 2 mg / mL or less.
　第二の液体に含まれるアミノ酸どうしの質量比（３種類以上含まれる場合は、任意の２種類間の質量比）は、例えば１０：９０～９０：１０の範囲であり、好ましくは２０：８０～８０：２０、より好ましくは３０：７０～７０：３０、更に好ましくは４０：６０～６０：４０である。 The mass ratio of amino acids contained in the second liquid (the mass ratio between any two types when 3 or more types are contained) is, for example, in the range of 10:90 to 90:10, preferably 20:80. 80:20, more preferably 30:70 to 70:30, and still more preferably 40:60 to 60:40.
　第二の液体の界面活性剤の濃度（複数種使用する場合はその総濃度）は、例えば０．０００１質量％～１質量％であり、好ましくは０．００１質量％～０．５質量％、より好ましくは０．００５質量％～０．５質量％、更に好ましくは０．００５質量％～０．３質量％、特に好ましくは０．００５質量％～０．１質量％である。 The concentration of the surfactant in the second liquid (the total concentration when plural kinds are used) is, for example, 0.0001% by mass to 1% by mass, preferably 0.001% by mass to 0.5% by mass, More preferably, it is 0.005% by mass to 0.5% by mass, still more preferably 0.005% by mass to 0.3% by mass, and particularly preferably 0.005% by mass to 0.1% by mass.
　第二の液体のアルコール濃度は、０．１質量％～２０質量％が好ましく、より好ましくは０．１質量％～１０質量％、更に好ましくは０．５質量％～１０質量％、特に好ましくは０．５質量％～５質量％である。 The alcohol concentration of the second liquid is preferably 0.1% by mass to 20% by mass, more preferably 0.1% by mass to 10% by mass, still more preferably 0.5% by mass to 10% by mass, particularly preferably. 0.5 mass% to 5 mass%.
　工程（Ｂ）において、製剤用添加物（賦形剤、崩壊剤、結合剤、滑沢剤、界面活性剤、緩衝剤、溶解補助剤、安定化剤、等張化剤など）を更に混合してもよい。また、アミノ酸以外の担体を混合してもよい。 In step (B), formulation additives (excipients, disintegrants, binders, lubricants, surfactants, buffers, solubilizers, stabilizers, tonicity agents, etc.) are further mixed. May be. A carrier other than amino acids may be mixed.
－工程（Ｃ）－
　工程（Ｃ）は、工程（Ｂ）で得た第二の液体を凍結乾燥する工程である。第二の液体を凍結乾燥する方法は特に制限されず、例えば、従来公知の凍結乾燥薬剤（例えば、用時溶解型の注射剤等）の製造に従来適用される凍結乾燥法でよい。 -Process (C)-
Step (C) is a step of freeze-drying the second liquid obtained in step (B). The method for freeze-drying the second liquid is not particularly limited, and may be, for example, a freeze-drying method that is conventionally applied to the production of a conventionally known freeze-dried drug (for example, an injection that is dissolved at the time of use).
　工程（Ｃ）は、単回投与分又は数回投与分の有効量（好ましくは単回投与分の有効量）の有効成分を含む第二の液体を、経肺投与用デバイスに装填可能な容器に充填し、そのまま凍結乾燥することが好ましい。 In the step (C), a container capable of charging a device for transpulmonary administration with a second liquid containing an active ingredient in an effective dose for single dose or several doses (preferably effective dose for single dose) It is preferable to fill in and freeze-dry as it is.
［経肺投与用医薬組成物］
　本発明の経肺投与用医薬組成物は、本発明の凍結乾燥組成物が気流によって粒子化されてなる粉末状の医薬組成物である。
　本発明の経肺投与用医薬組成物の一態様は、有効成分として、肺胞上皮幹細胞（好ましくはヒト肺胞上皮幹細胞）に対して分化誘導作用を有する化合物を含むものである。 [Pharmaceutical composition for transpulmonary administration]
The pharmaceutical composition for transpulmonary administration of the present invention is a powdery pharmaceutical composition obtained by granulating the lyophilized composition of the present invention into an air stream.
One aspect of the pharmaceutical composition for pulmonary administration of the present invention comprises a compound having an action of inducing differentiation of alveolar epithelial stem cells (preferably human alveolar epithelial stem cells) as an active ingredient.
　本発明の経肺投与用医薬組成物は、肺胞上皮幹細胞に対して分化誘導作用を有する化合物を含む場合、肺組織の損傷を呈する疾患の治療薬として使用し得る。当該疾患としては例えば、慢性閉塞性肺疾患（chronic obstructive pulmonary disease、ＣＯＰＤ）、肺気腫、慢性気管支炎、急性肺損傷（acute respiratory distress syndrome、ＡＲＤＳ）、肺線維症、肺癌、間質性肺炎、肺結核後遺症、じん肺などが挙げられる。
　したがって、本発明の経肺投与用医薬組成物によれば、肺組織の損傷を呈する疾患の治療方法が提供される。当該治療方法は、肺組織に損傷を有する患者（例えば上記の疾患の患者）に、本発明の経肺投与用医薬組成物を経肺投与することを含む、肺組織の損傷の治療方法である。
　本発明の経肺投与用医薬組成物の投与量は、疾患の種類；患者の症状、体重、年齢；有効成分として含まれる化合物の種類等に応じて選択される。 When the pharmaceutical composition for pulmonary administration of the present invention contains a compound having a differentiation-inducing action on alveolar epithelial stem cells, it can be used as a therapeutic agent for a disease exhibiting lung tissue damage. Examples of the disease include chronic obstructive pulmonary disease (COPD), emphysema, chronic bronchitis, acute lung distress syndrome (ARDS), pulmonary fibrosis, lung cancer, interstitial pneumonia, pulmonary tuberculosis. Examples include sequelae and pneumoconiosis.
Therefore, according to the pharmaceutical composition for transpulmonary administration of the present invention, a method for treating a disease exhibiting lung tissue damage is provided. The therapeutic method is a method for treating pulmonary tissue damage, comprising pulmonary administration of the pharmaceutical composition for pulmonary administration of the present invention to a patient having damage to pulmonary tissue (for example, a patient having the above-mentioned diseases). .
The dosage of the pharmaceutical composition for pulmonary administration of the present invention is selected according to the type of disease; patient symptom, body weight, age; type of compound contained as an active ingredient, and the like.
　本発明の経肺投与用医薬組成物は、調製用の凍結乾燥組成物が気流によって粒子化されてなる。
　気流の流量としては５Ｌ／ｍｉｎ～３００Ｌ／ｍｉｎ、より好ましくは１０Ｌ／ｍｉｎ～２００Ｌ／ｍｉｎ、更に好ましくは１０Ｌ／ｍｉｎ～１５０Ｌ／ｍｉｎ、特に好ましくは１０Ｌ／ｍｉｎ～１００Ｌ／ｍｉｎである。
　気流の流速としては１ｍ／ｓｅｃ～３００ｍ／ｓｅｃ、より好ましくは２ｍ／ｓｅｃ～２５０ｍ／ｓｅｃ、更に好ましくは５ｍ／ｓｅｃ～２００ｍ／ｓｅｃ、特に好ましくは１０ｍ／ｓｅｃ～１５０ｍ／ｓｅｃである。 The pharmaceutical composition for transpulmonary administration of the present invention is prepared by pulverizing a freeze-dried composition for preparation with an air stream.
The flow rate of the air flow is 5 L / min to 300 L / min, more preferably 10 L / min to 200 L / min, still more preferably 10 L / min to 150 L / min, and particularly preferably 10 L / min to 100 L / min.
The flow rate of the airflow is 1 m / sec to 300 m / sec, more preferably 2 m / sec to 250 m / sec, still more preferably 5 m / sec to 200 m / sec, and particularly preferably 10 m / sec to 150 m / sec.
　本発明の経肺投与用医薬組成物は、その調製用の凍結乾燥組成物が、比較的低い速度及び流量の気流によって粒子化され得るので、投与態様として例えば米国特許第７７３５４８５Ｂ２号明細書又は特許第４８２２７０９号公報に開示された経肺投与システムを採用することができる。
　即ち、凍結乾燥組成物を上記文献に開示された経肺投与用デバイスと共に提供すれば、当該デバイスの使用者が使用時に、デバイスに凍結乾燥組成物のバイアルを取り付け、患者の吸気により凍結乾燥組成物が経肺投与に適した粒子となり、そのまま吸入服用することができる。
　したがって、本発明の凍結乾燥組成物を、例えば上記経肺投与用デバイスと共に提供すれば、経肺投与用医薬組成物の経肺投与方法の一態様が提供される。当該経肺投与方法は、患者が本発明の経肺投与用医薬組成物を吸入する際に、本発明の凍結乾燥組成物に前記患者が気流をあてて粒子化して経肺投与に適した微粒子とする工程を含む。 The pharmaceutical composition for pulmonary administration of the present invention can be formed into particles by the lyophilized composition for its preparation by a relatively low velocity and flow rate of air flow, and therefore, as an administration mode, for example, US Pat. No. 7,735,485 B2 or Patent The transpulmonary administration system disclosed in Japanese Patent No. 4822709 can be employed.
That is, if the freeze-dried composition is provided together with the device for transpulmonary administration disclosed in the above document, the user of the device attaches a vial of the freeze-dried composition to the device at the time of use, and the freeze-dried composition is inhaled by the patient's inspiration. The product becomes particles suitable for transpulmonary administration and can be taken as it is by inhalation.
Therefore, if the lyophilized composition of the present invention is provided together with the above-mentioned device for pulmonary administration, for example, one embodiment of a method for pulmonary administration of the pharmaceutical composition for pulmonary administration is provided. In the pulmonary administration method, when the patient inhales the pharmaceutical composition for pulmonary administration of the present invention, the patient applies airflow to the freeze-dried composition of the present invention to form particles that are suitable for pulmonary administration. The process of including.
　本発明の経肺投与用医薬組成物において、全粒子に占める粒子径が５μｍ以下の粒子の割合は、１０％以上であることが好ましく、より好ましくは１５％以上、更に好ましくは２０％以上、更に好ましくは２５％以上、特に好ましくは３０％以上である。
　ここで粒子径は、幾何学的粒子径または空気力学的粒子径を意味する。上記の割合は、幾何学的粒子径については、全粒子に対する、粒子径５μｍ以下の粒子の割合（体積％）である。上記の割合は、空気力学的粒子径については、全粒子に含まれる薬物量（全薬物量）に対する、粒子径５μｍ以下の粒子に含まれる薬物量の割合（質量％）である。 In the pharmaceutical composition for pulmonary administration of the present invention, the proportion of particles having a particle size of 5 μm or less in the total particles is preferably 10% or more, more preferably 15% or more, still more preferably 20% or more, More preferably, it is 25% or more, and particularly preferably 30% or more.
Here, the particle diameter means a geometric particle diameter or an aerodynamic particle diameter. The above-mentioned ratio is the ratio (volume%) of particles having a particle diameter of 5 μm or less with respect to all the particles with respect to the geometric particle diameter. The above ratio is the ratio (mass%) of the amount of drug contained in particles having a particle diameter of 5 μm or less with respect to the amount of drug contained in all particles (total amount of drug).
　本発明の経肺投与医薬組成物において、全粒子に占める幾何学的粒子径が５μｍ以下の粒子の割合（体積％）は、１０％以上であることが好ましく、より好ましくは１５％以上、更に好ましくは２０％以上、更に好ましくは２５％以上、特に好ましくは３０％以上である。
　本発明の経肺投与医薬組成物において、全薬物量に対する空気力学的粒子径が５μｍ以下の粒子に含まれる薬物量の割合（質量％）は、１０％以上であることが好ましく、より好ましくは１５％以上、更に好ましくは２０％以上、更に好ましくは２５％以上、特に好ましくは３０％以上である。
　一般に、経肺的に投与された粒子が肺に到達し沈着するには、その粒子の空気力学的粒子径が５μｍ以下のミクロサイズであることが必要と言われている。多孔質の凍結乾燥組成物に由来する粒子の空気力学的粒子径は通常、幾何学的粒子径よりも小さいので、幾何学的粒子径５μｍ以下の粒子の割合が上記の範囲であれば、経肺投与に適した空気力学的粒子径の粒子が十分な割合で含まれることになる。 In the pulmonary administration pharmaceutical composition of the present invention, the proportion (volume%) of particles having a geometric particle size of 5 μm or less in the total particles is preferably 10% or more, more preferably 15% or more, and further Preferably it is 20% or more, More preferably, it is 25% or more, Most preferably, it is 30% or more.
In the pulmonary administration pharmaceutical composition of the present invention, the ratio (mass%) of the drug amount contained in particles having an aerodynamic particle size of 5 μm or less with respect to the total drug amount is preferably 10% or more, more preferably. It is 15% or more, more preferably 20% or more, further preferably 25% or more, and particularly preferably 30% or more.
In general, it is said that in order for particles administered transpulmonarily to reach and deposit in the lung, the particles must have an aerodynamic particle size of 5 μm or less. Since the aerodynamic particle size of the particles derived from the porous lyophilized composition is usually smaller than the geometric particle size, if the proportion of particles having a geometric particle size of 5 μm or less is within the above range, A sufficient proportion of particles of aerodynamic particle size suitable for pulmonary administration will be included.
　以下に実施例を挙げて、本発明をさらに具体的に説明する。以下の実施例に示す材料、使用量、割合、処理手順等は、本発明の趣旨を逸脱しない限り適宜変更することができる。したがって、本発明の範囲は以下に示す具体例により限定的に解釈されるべきものではない。 The present invention will be described more specifically with reference to the following examples. The materials, amounts used, ratios, processing procedures, and the like shown in the following examples can be changed as appropriate without departing from the spirit of the present invention. Therefore, the scope of the present invention should not be construed as being limited by the specific examples shown below.
　ＣＤ９０は肺胞上皮幹細胞のマーカーであり、ＡＱＰ－５はＩ型肺胞上皮細胞のマーカーであり、ＳＰ－ＡはＩＩ型肺胞上皮細胞のマーカーである。 CD90 is a marker for alveolar epithelial stem cells, AQP-5 is a marker for type I alveolar epithelial cells, and SP-A is a marker for type II alveolar epithelial cells.
［参考例１：ＡＴＲＡのヒト肺胞上皮幹細胞に対する分化誘導作用の検証］
　参考例１で使用したヒト肺胞上皮幹細胞は、肺疾患患者（肺癌又はＣＯＰＤ等における生検又は外科手術時の摘出組織）由来のヒト肺胞上皮幹細胞（Fujino N, at. al. Am. J. Respir. Cell Mol. Biol., 46, 422-430(2012)）である。当該ヒト肺胞上皮幹細胞は、１０％牛血清（ＦＢＳ）と１％の５０倍希釈ＭＥＭ（Amino Acids Solution）とを含むＤＭＥＭ培地で培養したマウス胎仔線維芽細胞の培養上清を、フィルター滅菌し調製した培養液を用いて、３７℃／５％ＣＯ２環境下で培養・維持した。 [Reference Example 1: Verification of differentiation-inducing action of ATRA on human alveolar epithelial stem cells]
Human alveolar epithelial stem cells used in Reference Example 1 were derived from human lung alveolar epithelial stem cells (Fujino N, at. Al. Am. J) derived from patients with lung disease (biopsy in lung cancer or COPD or surgical tissue removed during surgery). Respir. Cell Mol. Biol., 46, 422-430 (2012)). The human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
　前記培養液中のヒト肺胞上皮幹細胞（１×１０１～４ｃｅｌｌｓ／ｃｍ２）に、オールトランスレチノイン酸（ＡＴＲＡ）を０μＭ、０．０１μＭ、０．１μＭ、１μＭ又は１０μＭの濃度で曝露し、３７℃／５％ＣＯ２環境下で５日間、静置培養した。 All-trans retinoic acid (ATRA) is exposed to human alveolar epithelial stem cells (1 × 10 1-4 cells / cm 2 ) in the culture at a concentration of 0 μM, 0.01 μM, 0.1 μM, 1 μM, or 10 μM. The culture was stationary for 5 days in a 37 ° C./5% CO 2 environment.
　培養後の細胞をＰＢＳ（Phosphate Buffered Saline）で洗浄し、４％パラホルムアルデヒド－ＰＢＳ溶液を用いて固定した。この固定細胞にヤギ抗ヒトＣＤ９０抗体（Santa Cruz製）、ヤギ抗ヒトＡＱＰ－５抗体（Santa Cruz製）、ヤギ抗ヒトＳＰ－Ａ抗体（Santa Cruz製）を添加し反応させ、更にＦＩＴＣ標識ウサギ抗ヤギ抗体（Santa Cruz製）を添加し反応させた。そして、蛍光顕微鏡（ＢＺ－９０００、キーエンス社製）を用いて陽性細胞数を測定し、各マーカーについて陽性細胞の割合を算出した。その結果を表１に示す。 The cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution. To these fixed cells, goat anti-human CD90 antibody (SantaSCruz), goat anti-human AQP-5 antibody (Santa ヤ Cruz), goat anti-human SP-A antibody (Santa 添加 Cruz) are added and reacted. Anti-goat antibody (manufactured by Santa Cruz) was added and allowed to react. Then, the number of positive cells was measured using a fluorescence microscope (BZ-9000, manufactured by Keyence Corporation), and the percentage of positive cells was calculated for each marker. The results are shown in Table 1.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
　表１に明らかなとおり、ヒト肺胞上皮幹細胞にＡＴＲＡを接触させると、濃度依存的に、ＣＤ９０陽性細胞の割合が減少し、ＡＱＰ－５陽性細胞の割合とＳＰ－Ａ陽性細胞の割合が増加した。この結果は、ＡＴＲＡが、ヒト肺胞上皮幹細胞に対する分化誘導作用を有することを示している。 As is apparent from Table 1, when ATRA was contacted with human alveolar epithelial stem cells, the proportion of CD90 positive cells decreased and the proportion of AQP-5 positive cells and the proportion of SP-A positive cells increased in a concentration-dependent manner. did. This result indicates that ATRA has a differentiation-inducing action on human alveolar epithelial stem cells.
［Ａ］ＰＩ３キナーゼ阻害剤を含む医薬組成物
［実施例Ａ１：ＰＩ３Ｋ阻害剤のヒト肺胞上皮幹細胞に対する分化誘導作用の検証］
　実施例Ａ１で使用したヒト肺胞上皮幹細胞は、肺疾患患者（肺癌又はＣＯＰＤ等における生検又は外科手術時の摘出組織）由来のヒト肺胞上皮幹細胞（Fujino N, at. al. Am. J. Respir. Cell Mol. Biol., 46, 422-430(2012)）である。当該ヒト肺胞上皮幹細胞は、１０％牛血清（ＦＢＳ）と１％の５０倍希釈ＭＥＭ（Amino Acids Solution）とを含むＤＭＥＭ培地で培養したマウス胎仔線維芽細胞の培養上清を、フィルター滅菌し調製した培養液を用いて、３７℃／５％ＣＯ２環境下で培養・維持した。 [A] Pharmaceutical composition containing PI3 kinase inhibitor [Example A1: Verification of differentiation-inducing effect of PI3K inhibitor on human alveolar epithelial stem cells]
The human alveolar epithelial stem cells used in Example A1 are human alveolar epithelial stem cells (Fujino N, at. Al. Am. J) derived from patients with lung diseases (biopsy in lung cancer or COPD, or excised tissues at the time of surgery). Respir. Cell Mol. Biol., 46, 422-430 (2012)). The human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
　前記培養液中のヒト肺胞上皮幹細胞（１×１０１～４ｃｅｌｌｓ／ｃｍ２）に、ＰＩ３Ｋ阻害剤であるワートマニン（Wortmannin）を０μＭ、０．０１μＭ、０．１μＭ、１μＭ又は１０μＭの濃度で曝露し、３７℃／５％ＣＯ２環境下で５日間、静置培養した。 Human alveolar epithelial stem cells (1 × 10 1 to 4 cells / cm 2 ) in the culture medium are mixed with PI3K inhibitor Wortmannin at a concentration of 0 μM, 0.01 μM, 0.1 μM, 1 μM, or 10 μM. After exposure, the cells were statically cultured in a 37 ° C./5% CO 2 environment for 5 days.
　培養後の細胞をＰＢＳ（Phosphate Buffered Saline）で洗浄し、４％パラホルムアルデヒド－ＰＢＳ溶液を用いて固定した。この固定細胞にヤギ抗ヒトＣＤ９０抗体（Santa Cruz製）、ヤギ抗ヒトＡＱＰ－５抗体（Santa Cruz製）、ヤギ抗ヒトＳＰ－Ａ抗体（Santa Cruz製）を添加し反応させ、更にＦＩＴＣ標識ウサギ抗ヤギ抗体（Santa Cruz製）を添加し反応させた。そして、蛍光顕微鏡（ＢＺ－９０００、キーエンス社製）を用いて陽性細胞数を測定し、各マーカーについて陽性細胞の割合を算出した。その結果を表２に示す。 The cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution. To these fixed cells, goat anti-human CD90 antibody (SantaSCruz), goat anti-human AQP-5 antibody (Santa ヤ Cruz), goat anti-human SP-A antibody (Santa 添加 Cruz) are added and reacted. Anti-goat antibody (manufactured by Santa Cruz) was added and allowed to react. Then, the number of positive cells was measured using a fluorescence microscope (BZ-9000, manufactured by Keyence Corporation), and the percentage of positive cells was calculated for each marker. The results are shown in Table 2.
Figure JPOXMLDOC01-appb-T000002
Figure JPOXMLDOC01-appb-T000002
　表２に明らかなとおり、ヒト肺胞上皮幹細胞にＰＩ３Ｋ阻害剤を接触させると、濃度依存的に、ＣＤ９０陽性細胞の割合が減少し、ＡＱＰ－５陽性細胞の割合とＳＰ－Ａ陽性細胞の割合が増加した。この結果は、ＰＩ３Ｋ阻害剤が、ヒト肺胞上皮幹細胞に対する分化誘導作用を有することを示している。 As is apparent from Table 2, when a human alveolar epithelial stem cell was contacted with a PI3K inhibitor, the proportion of CD90 positive cells decreased in a concentration-dependent manner, and the proportion of AQP-5 positive cells and the proportion of SP-A positive cells increased. This result indicates that the PI3K inhibitor has a differentiation-inducing action on human alveolar epithelial stem cells.
　実施例Ａ１と、参考例１とを比較すると、ＰＩ３Ｋ阻害剤は、ＡＴＲＡよりも低濃度で、肺胞上皮幹細胞に対して強力な分化誘導作用を示した。例えば濃度０．１μＭどうしを比べると、ＰＩ３Ｋ阻害剤は、ＡＴＲＡの３倍以上のＩ型肺胞上皮細胞への分化誘導作用を示し、２倍以上のＩＩ型肺胞上皮細胞への分化誘導作用を示した。 When Example A1 and Reference Example 1 were compared, the PI3K inhibitor showed a strong differentiation-inducing action on alveolar epithelial stem cells at a lower concentration than ATRA. For example, when the concentrations of 0.1 μM are compared, the PI3K inhibitor has a differentiation-inducing action on type I alveolar epithelial cells that is 3 times or more of ATRA, and has a differentiation-inducing action on type II alveolar epithelial cells that is twice or more. showed that.
［実施例Ａ２：ＰＩ３Ｋ阻害剤のＣＯＰＤ動物モデルに対する効果の検証］
　ＣＯＰＤモデルマウス（エラスターゼ誘導性肺気腫マウス）を使って、ＰＩ３Ｋ阻害剤であるワートマニン（Wortmannin）の効果を確認する実験を行った。 [Example A2: Verification of effect of PI3K inhibitor on COPD animal model]
Using a COPD model mouse (elastase-induced emphysema mouse), an experiment was conducted to confirm the effect of Wortmannin, a PI3K inhibitor.
　６週齢のＩＣＲマウス（雄）に、ＰＢＳ５０μＬに懸濁したエラスターゼ（７．５Ｕ、Wako社製）を１週間にわたり週２回、ゾンデで経肺投与し、肺気腫モデルを作製した。続いて、ワートマニンを３週間にわたり週２回、各回１０μｇ／ｋｇの用量で、ゾンデで経肺投与した。最初のワートマニン投与から２１日後に両肺を摘出し、パラホルムアルデヒドで組織を固定し、クリオスタットで切片を作製し、ヘマトキシリン・エオジン染色（ＨＥ染色）を行った。 A 6-week-old ICR mouse (male) was administered pulmonary elastase (7.5 U, manufactured by Wako) suspended in 50 μL of PBS twice a week with a sonde for one week to prepare an emphysema model. Subsequently, wortmannin was transpulmonary administered twice a week for 3 weeks at a dose of 10 μg / kg each time with a sonde. Twenty-one days after the first administration of wortmannin, both lungs were removed, tissues were fixed with paraformaldehyde, sections were prepared with cryostat, and hematoxylin / eosin staining (HE staining) was performed.
　１匹のマウスの両肺の３野（左肺及び右肺の、上肺野、中肺野及び下肺野）の断面（合計６断面）のＨＥ染色スライドから、各断面５視野を顕微鏡（キーエンス社Biozero）で撮影し、画像解析ソフト（ＩｍａｇｅＪ（米国国立衛生研究所）及びＰｈｏｔｏｓｈｏｐ（Adobe社））を用いて解析した。気腫性病変の程度は、当業者に公知の方法であるThulbeckの方法に準じて平均肺胞壁間距離（mean linear intercept、Ｌｍ）を測定し評価した。その方法は、多数の無作為な線を引き、肺胞画像上にのせ、各無作為な線の肺胞壁によって分断された区間の長さの総和を求め、これを分断線の総数で除することによって求められる。１視野あたり肺胞隔壁による分断数を２００ 以上に設定し、これを各個体あたり３０視野（６断面×５視野）で測定し平均を計算し、各マウスの平均肺胞壁間距離を求め、各群の平均（個体数６匹）を算出し、ｔ検定で統計解析を行った。その結果を表３及び図１に示す。 From a HE-stained slide of a cross section (total 6 cross sections) of 3 fields (left lung and right lung, upper lung field, middle lung field and lower lung field) of both lungs of one mouse, 5 fields of each section were observed with a microscope ( The images were taken with Keyence Biozero and analyzed using image analysis software (ImageJ (National Institutes of Health, USA) and Photoshop (Adobe)). The degree of emphysematous lesions was evaluated by measuring the mean alveolar wall distance (mean linear intercept, Lm) according to the Thulbeck method, which is a method known to those skilled in the art. The method draws a number of random lines, places them on the alveolar image, finds the sum of the lengths of the sections separated by the alveolar walls of each random line, and divides this by the total number of broken lines. It is required by doing. The number of divisions by the alveolar septum per visual field is set to 200 mm or more, this is measured in 30 visual fields (6 cross sections x 5 visual fields) per individual, the average is calculated, and the average alveolar wall distance of each mouse is obtained, The average of each group (6 individuals) was calculated, and statistical analysis was performed by t-test. The results are shown in Table 3 and FIG.
Figure JPOXMLDOC01-appb-T000003
Figure JPOXMLDOC01-appb-T000003
　ＨＥ染色スライドの観察から、対照群（ワートマニンの投与なし）は、著明に肺胞腔が広がり、エラスターゼによって実験病理学的ＣＯＰＤ病変（顕著な肺気腫）ができていることが確認された。
　この対照群に比較して、ワートマニン投与群は、平均肺胞壁間距離が有意に低かった。平均肺胞壁間距離の低さは、肺胞が再生されたことを示しており、ワートマニン投与によって肺胞が再生されること、ワートマニンが肺組織損傷の治療に有効であること、が分かった。 From the observation of the HE-stained slide, it was confirmed that the alveolar space was remarkably expanded in the control group (without administration of wortmannin) and experimental pathological COPD lesions (remarkable emphysema) were formed by elastase.
Compared to this control group, the mean alveolar wall distance was significantly lower in the wortmannin administration group. The low average alveolar wall distance indicates that alveoli were regenerated, and it was found that wortmannin was regenerated and that wortmannin was effective in treating lung tissue damage. .
［Ｂ］ビタミンＤ受容体に作用する化合物を含む医薬組成物
　実施例Ｂ１及びＢ２で使用したヒト肺胞上皮幹細胞は、肺疾患患者（肺癌又はＣＯＰＤ等における生検又は外科手術時の摘出組織）由来のヒト肺胞上皮幹細胞（Fujino N, at. al. Am. J. Respir. Cell Mol. Biol., 46, 422-430(2012)）である。当該ヒト肺胞上皮幹細胞は、１０％牛血清（ＦＢＳ）と１％の５０倍希釈ＭＥＭ（Amino Acids Solution）とを含むＤＭＥＭ培地で培養したマウス胎仔線維芽細胞の培養上清を、フィルター滅菌し調製した培養液を用いて、３７℃／５％ＣＯ２環境下で培養・維持した。 [B] Pharmaceutical composition containing a compound that acts on vitamin D receptor Human alveolar epithelial stem cells used in Examples B1 and B2 are patients with lung disease (extracted tissue at the time of biopsy or surgery in lung cancer or COPD) Derived from human alveolar epithelial stem cells (Fujino N, at. Al. Am. J. Respir. Cell Mol. Biol., 46, 422-430 (2012)). The human alveolar epithelial stem cells are obtained by sterilizing the culture supernatant of mouse fetal fibroblasts cultured in DMEM medium containing 10% bovine serum (FBS) and 1% 50-fold diluted MEM (Amino Acids Solution). Using the prepared culture solution, it was cultured and maintained in a 37 ° C./5% CO 2 environment.
［実施例Ｂ１：１，２５－ジヒドロキシビタミンＤ３のヒト肺胞上皮幹細胞に対する分化誘導作用の検証（１）］
　前記培養液中のヒト肺胞上皮幹細胞（１×１０１～４ｃｅｌｌｓ／ｃｍ２）に、１，２５－ジヒドロキシビタミンＤ３を０μＭ、０．０１μＭ、０．１μＭ、１μＭ又は１０μＭの濃度で曝露し、３７℃／５％ＣＯ２環境下で５日間、静置培養した。 [Example B1: Verification of differentiation-inducing action of 1,25-dihydroxyvitamin D 3 on human alveolar epithelial stem cells (1)]
1,25-dihydroxyvitamin D 3 is exposed to human alveolar epithelial stem cells (1 × 10 1 to 4 cells / cm 2 ) in the culture at a concentration of 0 μM, 0.01 μM, 0.1 μM, 1 μM or 10 μM. Then, static culture was performed in a 37 ° C./5% CO 2 environment for 5 days.
　培養後の細胞をＰＢＳ（Phosphate Buffered Saline）で洗浄し、４％パラホルムアルデヒド－ＰＢＳ溶液を用いて固定した。この固定細胞にヤギ抗ヒトＣＤ９０抗体（Santa Cruz製）、ヤギ抗ヒトＡＱＰ－５抗体（Santa Cruz製）、ヤギ抗ヒトＳＰ－Ａ抗体（Santa Cruz製）を添加し反応させ、更にＦＩＴＣ標識ウサギ抗ヤギ抗体（Santa Cruz製）を添加し反応させた。そして、蛍光顕微鏡（ＢＺ－９０００、キーエンス社製）を用いて陽性細胞数を測定し、各マーカーについて陽性細胞の割合を算出した。その結果を表４に示す。 The cultured cells were washed with PBS (Phosphate Buffered Saline) and fixed with a 4% paraformaldehyde-PBS solution. To these fixed cells, goat anti-human CD90 antibody (SantaSCruz), goat anti-human AQP-5 antibody (Santa ヤ Cruz), goat anti-human SP-A antibody (Santa 添加 Cruz) are added and reacted. Anti-goat antibody (manufactured by Santa Cruz) was added and allowed to react. Then, the number of positive cells was measured using a fluorescence microscope (BZ-9000, manufactured by Keyence Corporation), and the percentage of positive cells was calculated for each marker. The results are shown in Table 4.
Figure JPOXMLDOC01-appb-T000004
Figure JPOXMLDOC01-appb-T000004
　表４に明らかなとおり、ヒト肺胞上皮幹細胞に１，２５－ジヒドロキシビタミンＤ３を接触させると、濃度依存的に、ＣＤ９０陽性細胞の割合が減少し、ＡＱＰ－５陽性細胞の割合とＳＰ－Ａ陽性細胞の割合が増加した。この結果は、１，２５－ジヒドロキシビタミンＤ３が、ヒト肺胞上皮幹細胞に対する分化誘導作用を有することを示している。 As is apparent from Table 4, when 1,25-dihydroxyvitamin D 3 was brought into contact with human alveolar epithelial stem cells, the proportion of CD90 positive cells decreased in a concentration-dependent manner, and the proportion of AQP-5 positive cells and SP- The proportion of A positive cells increased. This result indicates that 1,25-dihydroxyvitamin D 3 has a differentiation-inducing action on human alveolar epithelial stem cells.
　実施例Ｂ１と、参考例１とを比較すると、１，２５－ジヒドロキシビタミンＤ３は、ＡＴＲＡよりも低濃度で、肺胞上皮幹細胞に対して強力な分化誘導作用を示した。例えば濃度０．１μＭどうしを比べると、１，２５－ジヒドロキシビタミンＤ３は、ＡＴＲＡの７倍以上のＩ型肺胞上皮細胞への分化誘導作用を示し、３倍以上のＩＩ型肺胞上皮細胞への分化誘導作用を示した。 Example B1, is compared with the reference example 1, 1,25-dihydroxyvitamin D 3 is at a lower concentration than ATRA, it showed strong differentiation-inducing effect on alveolar epithelial stem cells. For example, when comparing concentrations of 0.1 μM, 1,25-dihydroxyvitamin D 3 has a differentiation-inducing action on type I alveolar epithelial cells that is 7 times or more of ATRA, and more than 3 times type II alveolar epithelial cells. Showed differentiation-inducing action.
［実施例Ｂ２：１，２５－ジヒドロキシビタミンＤ３のヒト肺胞上皮幹細胞に対する分化誘導作用の検証（２）］
　１，２５－ジヒドロキシビタミンＤ３の濃度を１０μＭにし、培養日数を０日、１日、２日、４日又は６日にした以外は、実施例Ｂ１と同様の実験を行った。その結果を表５に示す。
　培養日数０日の実験例では、１，２５－ジヒドロキシビタミンＤ３に曝露後直ちにＰＢＳで洗浄し、４％パラホルムアルデヒド－ＰＢＳ溶液で細胞を固定した。 [Example B2: Verification of differentiation-inducing action of 1,25-dihydroxyvitamin D 3 on human alveolar epithelial stem cells (2)]
An experiment similar to Example B1 was conducted, except that the concentration of 1,25-dihydroxyvitamin D 3 was 10 μM and the culture days were 0 days, 1 day, 2 days, 4 days or 6 days. The results are shown in Table 5.
In the experimental example with 0 days of culture, the cells were fixed with a 4% paraformaldehyde-PBS solution immediately after exposure to 1,25-dihydroxyvitamin D 3 and washed with PBS.
Figure JPOXMLDOC01-appb-T000005
Figure JPOXMLDOC01-appb-T000005
　表５に明らかなとおり、ヒト肺胞上皮幹細胞に１，２５－ジヒドロキシビタミンＤ３を接触させると、接触時間依存的に、ＣＤ９０陽性細胞の割合が減少し、ＡＱＰ－５陽性細胞の割合とＳＰ－Ａ陽性細胞の割合が増加した。この結果は、１，２５－ジヒドロキシビタミンＤ３が、ヒト肺胞上皮幹細胞に対する分化誘導作用を有することを示している。 As is apparent from Table 5, when 1,25-dihydroxyvitamin D 3 is contacted with human alveolar epithelial stem cells, the proportion of CD90 positive cells decreases, and the proportion of AQP-5 positive cells and SP -Increased proportion of A positive cells. This result indicates that 1,25-dihydroxyvitamin D 3 has a differentiation-inducing action on human alveolar epithelial stem cells.
［実施例Ｂ３：１，２５－ジヒドロキシビタミンＤ３のＣＯＰＤ動物モデルに対する効果の検証］
　ＣＯＰＤモデルマウス（エラスターゼ誘導性肺気腫マウス）を使って、１，２５－ジヒドロキシビタミンＤ３の効果を確認する実験を行った。 Example B3: 1,25- Verification of effect on COPD animal model dihydroxyvitamin D 3]
Experiments were conducted to confirm the effects of 1,25-dihydroxyvitamin D 3 using COPD model mice (elastase-induced emphysema mice).
　６週齢のＩＣＲマウス（雄）に、ＰＢＳ５０μＬに懸濁したエラスターゼ（７．５Ｕ、Wako社製）を１週間にわたり週２回、ゾンデで経肺投与し、肺気腫モデルを作製した。続いて、１，２５－ジヒドロキシビタミンＤ３を３週間にわたり週２回、各回１０μｇ／ｋｇの用量で、ゾンデで経肺投与した。最初の１，２５－ジヒドロキシビタミンＤ３投与から２１日後に両肺を摘出し、パラホルムアルデヒドで組織を固定し、クリオスタットで切片を作製し、ヘマトキシリン・エオジン染色（ＨＥ染色）を行った。 A 6-week-old ICR mouse (male) was subjected to pulmonary administration of elastase (7.5 U, manufactured by Wako) suspended in 50 μL of PBS twice a week with a sonde for one week to prepare an emphysema model. Subsequently, 1,25-dihydroxyvitamin D 3 twice a week for three weeks, at a dose of each time 10 [mu] g / kg, and transpulmonary administration at sonde. Twenty-one days after the first administration of 1,25-dihydroxyvitamin D 3 , both lungs were removed, tissues were fixed with paraformaldehyde, sections were prepared with cryostat, and hematoxylin / eosin staining (HE staining) was performed.
　１匹のマウスの両肺の３野（左肺及び右肺の、上肺野、中肺野及び下肺野）の断面（合計６断面）のＨＥ染色スライドから、各断面５視野を顕微鏡（キーエンス社Biozero）で撮影し、画像解析ソフト（ＩｍａｇｅＪ（米国国立衛生研究所）及びＰｈｏｔｏｓｈｏｐ（Adobe社））を用いて解析した。気腫性病変の程度は、当業者に公知の方法であるThulbeckの方法に準じて平均肺胞壁間距離（mean linear intercept、Ｌｍ）を測定し評価した。その方法は、多数の無作為な線を引き、肺胞画像上にのせ、各無作為な線の肺胞壁によって分断された区間の長さの総和を求め、これを分断線の総数で除することによって求められる。１視野あたり肺胞隔壁による分断数を２００ 以上に設定し、これを各個体あたり３０視野（６断面×５視野）で測定し平均を計算し、各マウスの平均肺胞壁間距離を求め、各群の平均（個体数６匹）を算出し、ｔ検定で統計解析を行った。その結果を表６及び図２に示す。 From a HE-stained slide of a cross section (total 6 cross sections) of 3 fields (left lung and right lung, upper lung field, middle lung field and lower lung field) of both lungs of one mouse, 5 fields of each section were observed with a microscope ( The images were taken with Keyence Biozero and analyzed using image analysis software (ImageJ (National Institutes of Health, USA) and Photoshop (Adobe)). The degree of emphysematous lesions was evaluated by measuring the mean alveolar wall distance (mean linear intercept, Lm) according to the Thulbeck method, which is a method known to those skilled in the art. The method draws a number of random lines, places them on the alveolar image, finds the sum of the lengths of the sections separated by the alveolar walls of each random line, and divides this by the total number of broken lines. It is required by doing. The number of divisions by the alveolar septum per visual field is set to 200 mm or more, this is measured in 30 visual fields (6 cross sections x 5 visual fields) per individual, the average is calculated, and the average alveolar wall distance of each mouse is obtained, The average of each group (6 individuals) was calculated, and statistical analysis was performed by t-test. The results are shown in Table 6 and FIG.
Figure JPOXMLDOC01-appb-T000006
Figure JPOXMLDOC01-appb-T000006
　ＨＥ染色スライドの観察から、対照群（１，２５－ジヒドロキシビタミンＤ３の投与なし）は、著明に肺胞腔が広がり、エラスターゼによって実験病理学的ＣＯＰＤ病変（顕著な肺気腫）ができていることが確認された。
　この対照群に比較して、１，２５－ジヒドロキシビタミンＤ３投与群は、平均肺胞壁間距離が有意に低かった。平均肺胞壁間距離の低さは、肺胞が再生されたことを示しており、１，２５－ジヒドロキシビタミンＤ３投与によって肺胞が再生されること、１，２５－ジヒドロキシビタミンＤ３が肺組織損傷の治療に有効であること、が分かった。 From the observation of the HE-stained slide, the control group (without administration of 1,25-dihydroxyvitamin D 3 ) has a markedly expanded alveolar space and experimental pathological COPD lesions (significant emphysema) caused by elastase. It was confirmed.
Compared to the control group, 1,25-dihydroxyvitamin D 3 administration group, the distance between the average alveolar wall were significantly lower. Low distances between the average alveolar wall shows that the alveoli is played, the alveoli is reproduced by the administration 1,25-dihydroxyvitamin D 3, the 1,25-dihydroxyvitamin D 3 It has been found to be effective in treating lung tissue damage.
［Ｃ］凍結乾燥組成物
　実施例Ｃ１～Ｃ４で使用したアミノ酸は、グリシン以外、Ｌ型である。 [C] Lyophilized composition The amino acids used in Examples C1 to C4 are in the L form other than glycine.
［実施例Ｃ１：ＡＴＲＡを含む凍結乾燥組成物の調製（１）］
［凍結乾燥組成物の調製］
　ＡＴＲＡ４ｍｇをエタノール１ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ２０を２０μＬ加え、ＡＴＲＡ溶液を調製した。
　別途、Ｐｈｅ、Ｌｅｕ、Ｉｌｅ及びＶａｌのアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は、Ｐｈｅは６ｍｇ／ｍＬ、Ｌｅｕ、Ｉｌｅ及びＶａｌは２．４ｍｇ／ｍＬとした。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表７に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。
　各分散液のエタノールの最終濃度は５質量％であった。各分散液の有効成分及びアミノ酸の最終量と界面活性剤の最終濃度とを表７に示した。１ｖｉａｌの分散液の液量は０．５ｍＬである。 [Example C1: Preparation of lyophilized composition containing ATRA (1)]
[Preparation of lyophilized composition]
4 mg of ATRA was dissolved in 1 mL of ethanol, and 20 μL of Tween 20 was added as a surfactant to prepare an ATRA solution.
Separately, amino acid aqueous solutions of Phe, Leu, Ile and Val were prepared. The amino acid concentration of the aqueous amino acid solution was 6 mg / mL for Phe, and 2.4 mg / mL for Leu, Ile, and Val.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 7, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
The final concentration of ethanol in each dispersion was 5% by weight. Table 7 shows the final amounts of the active ingredients and amino acids of each dispersion and the final concentration of the surfactant. The liquid volume of 1 vial dispersion is 0.5 mL.
　この分散液を、胴径１８ｍｍ・容量２ｍＬのガラス製容器（ｖｉａｌ）に０．５ｍＬ入れ、アセトン－ドライアイス下で急速凍結させ、その後、棚状凍結乾燥機（Ｆｒｅｅｚｏｎｅ Ｔｒｉａｄ、ＬＡＢＣＯＮＣＯ社製）を用いて凍結乾燥し、凍結乾燥組成物を得た。 0.5 mL of this dispersion is put into a glass container (vial) having a body diameter of 18 mm and a volume of 2 mL, and is rapidly frozen under acetone-dry ice. Thereafter, a shelf-like freeze dryer (Freezone Triad, manufactured by LABCONCO) is used. And lyophilized to obtain a lyophilized composition.
［凍結乾燥組成物の外観の評価］
　凍結乾燥組成物の外観を以下の評価基準に従って評価した。その結果を表７に示す。凍結乾燥組成物としては、５及び４が好ましい。
　５：形状が凍結乾燥組成物をなしており、その外観に偏り及びひびが認めらない。
　４：形状が凍結乾燥組成物をなしており、その外観に軽微なひびがある。
　３：形状が凍結乾燥組成物をなしており、その外観に偏り及び／又はひびがある。
　２：形状が凍結乾燥組成物をなしていない。
　１：凍結乾燥組成物ができていない又は極微量である。 [Evaluation of appearance of freeze-dried composition]
The appearance of the lyophilized composition was evaluated according to the following evaluation criteria. The results are shown in Table 7. As the lyophilized composition, 5 and 4 are preferable.
5: The shape is a lyophilized composition, and there is no bias or crack in its appearance.
4: The shape is a lyophilized composition, and its appearance has minor cracks.
3: The shape is a lyophilized composition, and the appearance is uneven and / or cracked.
2: The shape does not form a lyophilized composition.
1: The freeze-dried composition is not completed or is extremely small.
［凍結乾燥組成物の粒子化の評価］
　凍結乾燥組成物の一部を取り出し、レーザー回析式粒度分布測定装置（ＬＤＳＡ－３５００Ａ、東日コンピュータアプリケーションズ社製）の試料室に入れ、圧縮空気（速度６０ｍ／ｓｅｃ、流量７５０ｍｌ／ｓｅｃ）を試料室に送り込み凍結乾燥物を飛散させて、空気中での粒度分布（体積基準）を測定した。その粒度分布から、５μｍ以下の粒子割合（％）と、１０μｍ以下の粒子割合（％）を算出した。その結果を表７に示す。 [Evaluation of particle formation of freeze-dried composition]
A part of the freeze-dried composition is taken out and placed in the sample chamber of a laser diffraction particle size distribution analyzer (LDSA-3500A, manufactured by Tohnichi Computer Applications), and compressed air (speed 60 m / sec, flow rate 750 ml / sec) The lyophilized product was sent to the sample chamber and scattered, and the particle size distribution in air (volume basis) was measured. From the particle size distribution, a particle ratio (%) of 5 μm or less and a particle ratio (%) of 10 μm or less were calculated. The results are shown in Table 7.
Figure JPOXMLDOC01-appb-T000007
Figure JPOXMLDOC01-appb-T000007
　表７に明らかなとおり、実施例Ｃ１－１～Ｃ１－３の凍結乾燥組成物は、気流によって粒子化されて、粒子径５μｍ以下の粒子が２５％以上を占める粉末状の薬剤となった。この結果は、本発明の凍結乾燥組成物から、経肺投与に適した薬剤が調製されることを示している。 As is apparent from Table 7, the freeze-dried compositions of Examples C1-1 to C1-3 were granulated by an air flow to become a powdered drug in which particles having a particle diameter of 5 μm or less accounted for 25% or more. This result shows that a drug suitable for pulmonary administration is prepared from the lyophilized composition of the present invention.
［実施例Ｃ２：ＡＴＲＡを含む凍結乾燥組成物の調製（２）］
［実施例Ｃ２－１及び比較例Ｃ２－１］
　ＡＴＲＡ１ｍｇをエタノール０．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ８０を２．５μＬ加え、ＡＴＲＡ溶液を調製した。
　別途、ＰｈｅとＬｅｕのアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は、Ｐｈｅは４ｍｇ／ｍＬ、Ｌｅｕは１６ｍｇ／ｍＬとした。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表８に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。 [Example C2: Preparation of lyophilized composition containing ATRA (2)]
[Example C2-1 and Comparative Example C2-1]
1 mg of ATRA was dissolved in 0.25 mL of ethanol, and 2.5 μL of Tween 80 was added as a surfactant to prepare an ATRA solution.
Separately, amino acid aqueous solutions of Phe and Leu were prepared. The amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
［実施例Ｃ２－２］
　ＡＴＲＡ１ｍｇをエタノール０．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ８０を５μＬ加え、ＡＴＲＡ溶液を調製した。
　別途、ＰｈｅとＬｅｕのアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は、Ｐｈｅは４ｍｇ／ｍＬ、Ｌｅｕは１６ｍｇ／ｍＬとした。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表８に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。 [Example C2-2]
1 mg of ATRA was dissolved in 0.25 mL of ethanol, and 5 μL of Tween 80 was added as a surfactant to prepare an ATRA solution.
Separately, amino acid aqueous solutions of Phe and Leu were prepared. The amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
［比較例Ｃ２－２］
　ＡＴＲＡ５ｍｇをエタノール１．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ８０を２５μＬ加え、水で段階希釈し、ＡＴＲＡ溶液を調製した。
　別途、Ｐｈｅのアミノ酸水溶液（Ｐｈｅ濃度４ｍｇ／ｍＬ）を調製した。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表８に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。 [Comparative Example C2-2]
5 mg of ATRA was dissolved in 1.25 mL of ethanol, 25 μL of Tween 80 was added as a surfactant, and serially diluted with water to prepare an ATRA solution.
Separately, an amino acid aqueous solution of Phe (Phe concentration 4 mg / mL) was prepared.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
［実施例Ｃ２－３、及び比較例Ｃ２－３］
　ＡＴＲＡ５ｍｇをエタノール１．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ８０を２５μＬ加え、ＡＴＲＡ溶液を調製した。
　別途、ＰｈｅとＧｌｙのアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は、Ｐｈｅは８ｍｇ／ｍＬ、Ｇｌｙは８ｍｇ／ｍＬとした。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表８に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。 [Example C2-3 and Comparative Example C2-3]
5 mg of ATRA was dissolved in 1.25 mL of ethanol, and 25 μL of Tween 80 was added as a surfactant to prepare an ATRA solution.
Separately, amino acid aqueous solutions of Phe and Gly were prepared. The amino acid concentration of the aqueous amino acid solution was 8 mg / mL for Phe and 8 mg / mL for Gly.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 8, and the amount of the aqueous solution was adjusted with water to prepare a dispersion in which ATRA was dispersed.
　各分散液のエタノールの最終濃度は５質量％であった。各分散液の有効成分及びアミノ酸の最終量と界面活性剤の最終濃度とを表８に示した。１ｖｉａｌの分散液の液量は０．５ｍＬである。 The final concentration of ethanol in each dispersion was 5% by mass. Table 8 shows the final amounts of the active ingredients and amino acids of each dispersion and the final concentration of the surfactant. The liquid volume of 1 vial dispersion is 0.5 mL.
　この分散液から実施例Ｃ１と同様の方法で凍結乾燥組成物を得て、外観の評価及び粒子化の評価を行った。その結果を表８に示す。 From this dispersion, a freeze-dried composition was obtained in the same manner as in Example C1, and the appearance and particle formation were evaluated. The results are shown in Table 8.
Figure JPOXMLDOC01-appb-T000008
Figure JPOXMLDOC01-appb-T000008
　表８に明らかなとおり、実施例Ｃ２－１～Ｃ２－３の凍結乾燥組成物は、気流によって粒子化されて、粒子径５μｍ以下の粒子が１５％以上を占める粉末状の薬剤となった。この結果は、本発明の凍結乾燥組成物から、経肺投与に適した薬剤が調製されることを示している。 As is apparent from Table 8, the freeze-dried compositions of Examples C2-1 to C2-3 were granulated by an air flow to become a powdered drug in which particles having a particle diameter of 5 μm or less accounted for 15% or more. This result shows that a drug suitable for pulmonary administration is prepared from the lyophilized composition of the present invention.
［実施例Ｃ３：ＡＴＲＡを含む凍結乾燥組成物の調製（３）］
　ＡＴＲＡ５ｍｇをエタノール１．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ８０を２５μＬ加え、ＡＴＲＡ溶液を調製した。
　別途、表９に記載の種類のアミノ酸のアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は８ｍｇ／ｍＬとした。
　そして、ＡＴＲＡ溶液にアミノ酸水溶液を、アミノ酸の最終量が表９に記載した量となるように添加し、水で液量を調節し、ＡＴＲＡが分散した分散液を調製した。
　各分散液のエタノールの最終濃度は５質量％であった。各分散液の有効成分及びアミノ酸の最終量と界面活性剤の最終濃度とを表９に示した。１ｖｉａｌの分散液の液量は０．５ｍＬである。 [Example C3: Preparation of lyophilized composition containing ATRA (3)]
5 mg of ATRA was dissolved in 1.25 mL of ethanol, and 25 μL of Tween 80 was added as a surfactant to prepare an ATRA solution.
Separately, amino acid aqueous solutions of amino acids described in Table 9 were prepared. The amino acid concentration of the aqueous amino acid solution was 8 mg / mL.
Then, an aqueous amino acid solution was added to the ATRA solution so that the final amount of amino acid was as shown in Table 9, and the liquid volume was adjusted with water to prepare a dispersion in which ATRA was dispersed.
The final concentration of ethanol in each dispersion was 5% by weight. Table 9 shows the final amounts of the active ingredients and amino acids and the final concentration of the surfactant in each dispersion. The liquid volume of 1 vial dispersion is 0.5 mL.
　この分散液から実施例Ｃ１と同様の方法で凍結乾燥組成物を得て、外観の評価及び粒子化の評価を行った。その結果を表９に示す。 From this dispersion, a freeze-dried composition was obtained in the same manner as in Example C1, and the appearance and particle formation were evaluated. The results are shown in Table 9.
Figure JPOXMLDOC01-appb-T000009
Figure JPOXMLDOC01-appb-T000009
　表９に明らかなとおり、実施例Ｃ３－１～Ｃ３－４の凍結乾燥組成物は、気流によって粒子化されて、粒子径５μｍ以下の粒子が１０％以上を占める粉末状の薬剤となった。この結果は、本発明の凍結乾燥組成物から、経肺投与に適した薬剤が調製されることを示している。 As is apparent from Table 9, the lyophilized compositions of Examples C3-1 to C3-4 were granulated by an air flow to become a powdered drug in which particles having a particle diameter of 5 μm or less accounted for 10% or more. This result shows that a drug suitable for pulmonary administration is prepared from the lyophilized composition of the present invention.
［実施例Ｃ４：１，２５－ジヒドロキシビタミンＤ３を含む凍結乾燥組成物の調製］
　１，２５－ジヒドロキシビタミンＤ３０．１ｍｇをエタノール０．２５ｍＬに溶解させ、界面活性剤としてＴｗｅｅｎ２０を５μＬ加え溶解した。
　別途、Ｐｈｅ及びＬｅｕのアミノ酸水溶液をそれぞれ調製した。アミノ酸水溶液のアミノ酸濃度は、Ｐｈｅは４ｍｇ／ｍＬ、Ｌｅｕは１６ｍｇ／ｍＬとした。
　そして、１，２５－ジヒドロキシビタミンＤ３溶液にアミノ酸水溶液を、アミノ酸の最終量が表１０に記載した量となるように添加し、水で液量を調節し、１，２５－ジヒドロキシビタミンＤ３が溶解した溶解液（５ｍＬ）を調製した。
　各分散液のエタノールの最終濃度は５質量％であった。各分散液の有効成分及びアミノ酸の最終量と界面活性剤の最終濃度とを表１０に示した。１ｖｉａｌの分散液の液量は０．５ｍＬである。 Example C4: 1,25- Preparation of lyophilized compositions that comprise the dihydroxyvitamin D 3]
0.1 mg of 1,25-dihydroxyvitamin D 3 was dissolved in 0.25 mL of ethanol, and 5 μL of Tween 20 was added as a surfactant and dissolved.
Separately, amino acid aqueous solutions of Phe and Leu were prepared. The amino acid concentration of the aqueous amino acid solution was 4 mg / mL for Phe and 16 mg / mL for Leu.
Then, an aqueous amino acid solution is added to the 1,25-dihydroxyvitamin D 3 solution so that the final amount of the amino acid is as shown in Table 10, and the liquid volume is adjusted with water, and 1,25-dihydroxyvitamin D 3 is added. A solution (5 mL) in which was dissolved was prepared.
The final concentration of ethanol in each dispersion was 5% by weight. Table 10 shows the final amounts of active ingredients and amino acids and the final concentration of the surfactant in each dispersion. The liquid volume of 1 vial dispersion is 0.5 mL.
　この分散液から実施例Ｃ１と同様の方法で凍結乾燥組成物を得て、外観の評価及び粒子化の評価を行った。その結果を表１０に示す。 From this dispersion, a freeze-dried composition was obtained in the same manner as in Example C1, and the appearance and particle formation were evaluated. The results are shown in Table 10.
Figure JPOXMLDOC01-appb-T000010
Figure JPOXMLDOC01-appb-T000010
　表１０に明らかなとおり、実施例Ｃ４の凍結乾燥組成物は、気流によって粒子化されて、粒子径５μｍ以下の粒子が２０％以上を占める粉末状の薬剤となった。この結果は、本発明の凍結乾燥組成物から、経肺投与に適した薬剤が調製されることを示している。 As is apparent from Table 10, the freeze-dried composition of Example C4 was granulated by an air flow, and became a powdered drug in which particles having a particle diameter of 5 μm or less accounted for 20% or more. This result shows that a drug suitable for pulmonary administration is prepared from the lyophilized composition of the present invention.
　２０１２年１２月２１日に出願された日本国特許出願２０１２－２８００７０号の開示は、その全体が参照により本明細書に取り込まれる。
　本明細書に記載された全ての文献、特許出願、及び技術規格は、個々の文献、特許出願、及び技術規格が参照により取り込まれることが具体的かつ個々に記された場合と同程度に、本明細書中に参照により取り込まれる。 The disclosure of Japanese Patent Application No. 2012-280070 filed on December 21, 2012 is incorporated herein by reference in its entirety.
All documents, patent applications, and technical standards mentioned in this specification are to the same extent as if each individual document, patent application, and technical standard were specifically and individually stated to be incorporated by reference, Incorporated herein by reference.

Claims (17)

  1. 　ＰＩ３キナーゼ阻害剤を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。 A differentiation inducer that acts on alveolar epithelial stem cells, containing a PI3 kinase inhibitor as an active ingredient.
  2. 　ＰＩ３キナーゼ阻害剤を有効成分として含む、肺胞再生を誘導する医薬組成物。 A pharmaceutical composition for inducing alveolar regeneration comprising a PI3 kinase inhibitor as an active ingredient.
  3. 　ＰＩ３キナーゼ阻害剤を有効成分として含む、肺組織の損傷治療用の医薬組成物。 A pharmaceutical composition for treating lung tissue damage, comprising a PI3 kinase inhibitor as an active ingredient.
  4. 　ビタミンＤ受容体に作用する化合物を有効成分として含む、肺胞上皮幹細胞に作用する分化誘導剤。 A differentiation-inducing agent that acts on alveolar epithelial stem cells, containing a compound that acts on vitamin D receptor as an active ingredient.
  5. 　ビタミンＤ受容体に作用する化合物を有効成分として含む、肺胞再生を誘導する医薬組成物。 A pharmaceutical composition for inducing alveolar regeneration, comprising a compound that acts on a vitamin D receptor as an active ingredient.
  6. 　ビタミンＤ受容体に作用する化合物を有効成分として含む、肺組織の損傷治療用の医薬組成物。 A pharmaceutical composition for treating lung tissue damage, comprising a compound that acts on a vitamin D receptor as an active ingredient.
  7. 　水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤とを含み、
    　気流によって粒子化された際に、粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。     Containing a poorly water-soluble drug, at least two amino acids, and a surfactant,
    A freeze-dried composition in which the proportion of particles having a particle size of 5 μm or less becomes 10% or more when formed into particles by an air stream.
  8. 　水に難溶性の薬物と、少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物であり、
    　気流によって粒子化された際に、粒子径５μｍ以下の粒子の割合が１０％以上となる凍結乾燥組成物。     A freeze-dried composition of a composition comprising a poorly water-soluble drug, at least two amino acids, a surfactant, an alcohol, and water;
    A freeze-dried composition in which the proportion of particles having a particle size of 5 μm or less becomes 10% or more when formed into particles by an air stream.
  9. 　水に難溶性の薬物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤とを含む凍結乾燥組成物。
    　・第１群：フェニルアラニン、トリプトファン
    　・第２群：イソロイシン、ロイシン、バリン
    　・第３群：アラニン、グリシン     A freeze-dried composition comprising a poorly water-soluble drug, at least two amino acids selected from the first group, the second group, and the third group described below, and a surfactant, and a surfactant.
    ・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
  10. 　水に難溶性の薬物と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、アルコールと、水とを含む組成物の凍結乾燥組成物。
    　・第１群：フェニルアラニン、トリプトファン
    　・第２群：イソロイシン、ロイシン、バリン
    　・第３群：アラニン、グリシン     A composition comprising a poorly water-soluble drug, at least two amino acids selected from the following first group, second group and third group, a plurality of groups, a surfactant, alcohol, and water Freeze-dried composition.
    ・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
  11. 　前記水に難溶性の薬物が、肺胞上皮幹細胞に対して分化誘導作用を有する化合物である請求項７～請求項１０のいずれか１項に記載の凍結乾燥組成物。 The freeze-dried composition according to any one of claims 7 to 10, wherein the water-insoluble drug is a compound having a differentiation-inducing action on alveolar epithelial stem cells.
  12. 　前記肺胞上皮幹細胞に対して分化誘導作用を有する化合物が、ビタミンＡ、ビタミンＡ誘導体、プロビタミンＡ、ビタミンＤ受容体に作用する化合物、及びＰＩ３キナーゼ阻害剤から選ばれた少なくとも１種である請求項１１に記載の凍結乾燥組成物。 The compound having differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors. The lyophilized composition according to claim 11.
  13. 　水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、
    　前記第一の液体と、少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、
    　前記第二の液体を凍結乾燥する工程と
    を含む凍結乾燥組成物の製造方法。     A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol;
    Mixing the first liquid, at least two types of amino acids, a surfactant, and water to prepare a second liquid;
    A method of producing a freeze-dried composition comprising the step of freeze-drying the second liquid.
  14. 　水に難溶性の薬物を、アルコールを含む溶媒に溶解させて第一の液体を調製する工程と、
    　前記第一の液体と、下記の第１群、第２群及び第３群から複数群にまたがって選ばれた少なくとも２種類のアミノ酸と、界面活性剤と、水とを混合して第二の液体を調製する工程と、
    　前記第二の液体を凍結乾燥する工程と
    を含む凍結乾燥組成物の製造方法。
    　・第１群：フェニルアラニン、トリプトファン
    　・第２群：イソロイシン、ロイシン、バリン
    　・第３群：アラニン、グリシン     A step of preparing a first liquid by dissolving a poorly water-soluble drug in a solvent containing alcohol;
    The first liquid, at least two kinds of amino acids selected from the following first group, second group and third group, a plurality of groups, a surfactant and water are mixed to form a second Preparing a liquid;
    A method of producing a freeze-dried composition comprising the step of freeze-drying the second liquid.
    ・ Group 1: phenylalanine, tryptophan ・ Group 2: isoleucine, leucine, valine ・ Group 3: alanine, glycine
  15. 　前記水に難溶性の薬物が、肺胞上皮幹細胞に対して分化誘導作用を有する化合物である請求項１３又は請求項１４に記載の凍結乾燥組成物の製造方法。 The method for producing a lyophilized composition according to claim 13 or 14, wherein the poorly water-soluble drug is a compound having a differentiation-inducing action on alveolar epithelial stem cells.
  16. 　前記肺胞上皮幹細胞に対して分化誘導作用を有する化合物が、ビタミンＡ、ビタミンＡ誘導体、プロビタミンＡ、ビタミンＤ受容体に作用する化合物、及びＰＩ３キナーゼ阻害剤から選ばれた少なくとも１種である請求項１５に記載の凍結乾燥組成物の製造方法。 The compound having differentiation-inducing action on alveolar epithelial stem cells is at least one selected from vitamin A, vitamin A derivatives, provitamin A, compounds acting on vitamin D receptors, and PI3 kinase inhibitors. The method for producing a lyophilized composition according to claim 15.
  17. 　請求項７～請求項１２のいずれか１項に記載の凍結乾燥組成物を気流によって粒子化した経肺投与用医薬組成物。 A pharmaceutical composition for transpulmonary administration, wherein the freeze-dried composition according to any one of claims 7 to 12 is granulated by an air stream.
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