WO2023237626A1 - Specialised pro-resolution lipid mediator (spm) compounds for use in the treatment of cystic fibrosis and aspergillus fumigatus infection in patient suffering from cystic fibrosis - Google Patents

Specialised pro-resolution lipid mediator (spm) compounds for use in the treatment of cystic fibrosis and aspergillus fumigatus infection in patient suffering from cystic fibrosis Download PDF

Info

Publication number
WO2023237626A1
WO2023237626A1 PCT/EP2023/065286 EP2023065286W WO2023237626A1 WO 2023237626 A1 WO2023237626 A1 WO 2023237626A1 EP 2023065286 W EP2023065286 W EP 2023065286W WO 2023237626 A1 WO2023237626 A1 WO 2023237626A1
Authority
WO
WIPO (PCT)
Prior art keywords
compound
resolvin
cftr
lipoxin
spm
Prior art date
Application number
PCT/EP2023/065286
Other languages
French (fr)
Inventor
Valérie URBACH
Khadeeja Adam SY
Françoise BOTTEREL
Original Assignee
Institut National de la Santé et de la Recherche Médicale
Assistance Publique-Hôpitaux De Paris (Aphp)
Université Paris Est Créteil Val De Marne
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Institut National de la Santé et de la Recherche Médicale, Assistance Publique-Hôpitaux De Paris (Aphp), Université Paris Est Créteil Val De Marne filed Critical Institut National de la Santé et de la Recherche Médicale
Publication of WO2023237626A1 publication Critical patent/WO2023237626A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—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
    • A61K31/202—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 having three or more double bonds, e.g. linolenic
    • 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
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/10—Antimycotics

Definitions

  • SPECIALISED PRO-RESOLUTION LIPID MEDIATOR COMPOUNDS FOR USE IN THE TREATMENT OF CYSTIC FIBROSIS AND ASPERGILLUS FUMIGATUS INFECTION IN PATIENT SUFFERING FROM CYSTIC FIBROSIS
  • the invention is in the field of pneumonology. More particularly, the invention relates to methods and compositions for treating cystic fibrosis airway disease, and in particular repairing/prevent airway epithelial damage. The invention relates also to methods for treating Aspergillus fumigatus infection in patients suffering from cystic fibrosis.
  • Cystic fibrosis is an autosomal recessive genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene (1). More than 2,100 different mutations have been described affecting protein production or activity.
  • the CFTR chloride channel mainly expressed in epithelia, regulates transepithelial salt and water fluxes.
  • impaired chloride ion efflux coupled with sodium ion hyperabsorption results in epithelial surface dehydration and thick and tenacious mucus that obstruct the lungs, pancreatic ducts, the biliary tract, the intestine, and the male reproductive tract.
  • Chronic lung infections are CF’s most common clinical manifestation, leading to pulmonary sustained inflammation, progressive tissue destruction and loss of function, and elevated morbidity and mortality.
  • Cystic fibrosis is characterized by a chronic microbial infection of the respiratory tree associated with persistent inflammation.
  • Aspergillus fumigatus is the predominant filamentous fungus isolated from respiratory cultures of patients with CF and whose prevalence appears to be increasing. Aspergillus is associated with progressive structural lung injury and/or impaired lung function. Aspergillus fumigatus poses a serious health threat to humans by causing numerous invasive infections and a notable mortality in humans, especially in immunocompromised patient. Aspergillus fumigatus is thus included in the critical group (i.e pathogens ranked for perceived public health importance) on the fungal priority pathogens list developed by World health organization.
  • the inflammatory process is normally self-regulated by an active resolution phase, involving specialized lipid mediators of the resolution of inflammation (SPMs) such as lipoxins (LX), resolvins (Rv), protecins (Pd) and maresins (MaR).
  • SPMs lipid mediators of the resolution of inflammation
  • LX lipoxins
  • Rv resolvins
  • Pd protecins
  • MaR maresins
  • the present invention relates to methods and pharmaceutical compositions for the treatment of cystic fibrosis.
  • the present invention also relates to methods and pharmaceutical compositions for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis.
  • the present invention is defined by the claims.
  • SPMs pro-resolution lipid mediators
  • the invention relates to a method for treating cystic fibrosis in patient in need thereof comprising administering to said patient a therapeutically effective amount of at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound RvD5, lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPMs specialized lipid mediators of the resolution of inflammation
  • the invention relates to at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound RvD5, lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4), for use for treating cystic fibrosis in patient in need thereof
  • SPMs specialized lipid mediators of the resolution of inflammation
  • the invention relates to at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound RvD5 for use for treating cystic fibrosis in patient in need thereof
  • SPMs specialized lipid mediators of the resolution of inflammation
  • the terms “subject” or “patient” denote a mammal, such as a rodent, a feline, a canine, pig, ferret, rat, mice, rabbit and a primate.
  • a subject according to the invention refers to any subject (preferably human) afflicted or at risk to be afflicted with cystic fibrosis.
  • the method of the invention may be performed for any type of cystic fibrosis revised in the World Health Organization Classification of cystic fibrosis and selected from the E84 group: mucoviscidosis or Cystic fibrosis with pulmonary manifestations.
  • the subject according to the invention is a human.
  • the subject according to the invention is a girl or a boy.
  • the subject according to the invention is an adult. In some embodiments, the subject according to the invention is a child (human being between the stages of birth and puberty), a teenager (human being between the stages of puberty to adulthood) or an elderly person (human being after the puberty).
  • CF cystic fibrosis transmembrane conductance regulator
  • CFTR protein refers to the CFTR protein of 1480 amino acids, also called Cystic Fibrosis Transmembrane conductance Regulator.
  • the CFTR protein is a chloride (C1-) channel found in the membranes of secretory tissues as intestinal and respiratory mucosa.
  • the CFTR protein is represented by the NCBI reference sequence: P13569.3 (SEQ: ID NO: 1)
  • CFTR gene refers to the CFTR gene which is located on chromosome 7 and which may be found in NCBI GenBank locus AC000111 and AC000061, the contents of which are incorporated herein in their entirety by reference.
  • the cDNA for the CFTR gene is found in Audrezet et al., Hum. Mutat. (2004) 23 (4), 343-357.
  • a nucleic acid sequence for human CFTR is represented by SEQ ID NO: 2.
  • mutation has its general meaning in the art and refers to any detectable change in genetic material, e.g., DNA, RNA, cDNA, or any process, mechanism, or result of such a change. This includes gene mutations, in which the structure (e.g., DNA sequence) of a gene is altered, any gene or DNA arising from any mutation process, and any expression product (e.g., protein or enzyme) expressed by a modified gene or DNA sequence. Mutations include deletion, insertion or substitution of one or more nucleotides.
  • the mutation may occur in the coding region of a gene (i.e., in exons), in introns, or in the regulatory regions (e.g., enhancers, response elements, suppressors, signal sequences, polyadenylation sequences, promoters) of the gene.
  • a mutation is identified in a subject by comparing the sequence of a nucleic acid or polypeptide expressed by said subject with the corresponding nucleic acid or polypeptide expressed in a control population.
  • the mutation may be a “missense” mutation, where it replaces one amino acid with another in the gene product, or a “non-sense” mutation , where it replaces an amino acid codon with a stop codon.
  • “>” indicates a substitution at DNA level; (underscore) indicates a range of affected residues, separating the first and last residue affected; “del” indicates a deletion, “dup” indicates a duplication; “ins” indicates a insertion, “inv” indicates an inversion and “con” indicates a conversion. More particularly, “X” denotes that an amino acid is changed to a stop codon (X).
  • the term “homozygous” refers to an individual possessing two copies of the same allele.
  • the term “homozygous mutant” refers to an individual possessing two copies of the same allele, such allele being characterized as the mutant form of a gene.
  • heterozygous refers to an individual possessing two different alleles of the same gene, i.e., an individual possessing two different copies of an allele, such alleles are characterized as mutant forms of a gene.
  • the subject harbors at least one mutation in the CFTR gene.
  • the CFTR gene mutations were classified into six classes according to their resulting damaging effect on the protein (Elbom JS, 2016).
  • the class I mutations contribute to the formation of proteins with incomplete length and usually involve the complete loss of its activity (e.g.,W1282X, 1717-1G->A, G542X, R553X, 2183AA>G).
  • Mutation in the class II lead to abnormal maturation of proteins in the endoplasmic reticulum and Golgi apparatus. The effect of these mutations is premature degradation of the CFTR protein.
  • CFTR protein does not reach the cell membrane where it should perform its function (e.g., F508del, 2184delA).
  • the gene product having mutations of class III is properly synthesized, transported, and incorporated into the cell membrane, but has decreased activity caused by abnormal regulation of the protein. These mutations are frequently situated within one of the nucleotide binding domains, (e.g., G551D, R560T). Mutations of class IV cause anomalies in the structure of the transmembrane protein and thereby reduce the conduction of the chloride channel (e.g., R117H, R334W). Mutations altering the stability of the mRNA coding for the CFTR protein represent a class V of the mutations of the CFTR gene (e.g., 2789+5G->A, A455E).
  • Mutations in class VI lead to a decrease of the CFTR protein stability (e.g., 4326delTC, Glnl412X, 4279insA) (Harriet C., Kristin E. T., Olivier T., et al. Translating the genetics of cystic fibrosis to personalized medicine. Transl Res 2016; 168 40-49 and Elbom JS. Cystic fibrosis. Seminar. 2016).
  • CFTR protein stability e.g., 4326delTC, Glnl412X, 4279insA
  • the subject harbors at least one allelic mutation selected from class
  • the subject harbors at least a mutation of class I in the first allele and at least a mutation of class II in the second allele. In a particular embodiment, the subject harbors at least a mutation of class II in the first allele and at least a mutation of class II in the second allele.
  • the subject harbors at least a F508del mutation in the CFTR gene.
  • therapeutic regimen is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy.
  • a therapeutic regimen may include an induction regimen and a maintenance regimen.
  • the phrase “induction regimen” or “induction period” refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease.
  • the general goal of an induction regimen is to provide a high level of drug to a subject during the initial period of a treatment regimen.
  • An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both.
  • maintenance regimen refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a subject during treatment of an illness, e.g., to keep the subject in remission for long periods of time (months or years).
  • a maintenance regimen may employ continuous therapy (e.g., administering a drug at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of particular predetermined criteria [e.g., disease manifestation, etc.]).
  • administering refers to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., at least one SPMs of the invention) into the subject, such as by, intravenous, intramuscular, enteral, subcutaneous, parenteral, systemic, local, spinal, nasal, topical or epidermal administration (e.g., by injection or infusion).
  • a disease, or a symptom thereof is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof.
  • administration of the substance typically occurs before the onset of the disease or symptoms thereof.
  • the drug is administered to the subject using any suitable method that enables the drug to reach the lungs, pancreas, intestine, skin, reproductive system.
  • the drug administered to the patients systemically (i.e., via systemic administration).
  • the drug is administered to the subject such that it enters the circulatory system and is distributed throughout the body.
  • the drug is administered to the subject by local administration.
  • the at least one SPMs of the invention is administered by oral administration.
  • the at least one SPMs of the invention is administered by nasal administration.
  • the at least one SPMs of the invention is administered by inhalation.
  • a “therapeutically effective amount” is intended for a minimal amount of active agent which is necessary to impart therapeutic benefit to a subject.
  • a “therapeutically effective amount” to a subject is such an amount which induces, ameliorates or otherwise causes an improvement in the pathological symptoms, disease progression or physiological conditions associated with or resistance to succumbing to a disorder. It will be understood that the total daily usage of the compounds of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated.
  • a medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient.
  • An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
  • specialized lipid mediators of the resolution of inflammation or “specialized pro-resolving mediators (SPMs)” has its general meaning in the art and refers to a class of cell signaling molecules formed in the organism by the metabolism of polyunsaturated fatty acids (PUFA) by a combination of lipoxygenase, cyclooxygenase, and cytochrome P450 monooxygenase enzymes.
  • SPM are metabolites of arachidonic acid (AA), eicosapentaenoic acid (EP A), docosahexaenoic acid (DHA), or n-3 DPA (i.e.
  • SPM serotonin-associated phagocytosis .
  • LX lipoxins
  • Rv resolvins
  • PD protectins
  • MaR maresins
  • SPMs includes protectin DI (PD1), resolvin El (RvEl), resolvin E2 (RvE2), resolvin E3 (RvE3), resolvin E4 (RvE4), resolvin DI (RvDl), resolvin D2 (RvD2), resolvin D3 (RvD3), resolvin D4 (RvD4), resolvin D5 (RvD5), lipoxin A4 (LXA4) and lipoxin B4 (LXB4).
  • RvEl has the following structure and CAS number: 552830-51-0:
  • resolvin DI compound or “resolvin DI” or “RvDl” refers to one of the potent lipid mediators derived from both eicosapentaenoic acid (EP A) and docosahexaenoic acid (DHA).
  • Resolvin DI (RvDl) is produced physiologically from the sequential oxygenation of DHA by 15- and 5-lipoxygenase. In addition to being antiinflammatory, resolvins promote the resolution of the inflammatory response back to a noninflamed state.
  • RvDl has the following structure and CAS number: 872993-05-0:
  • resolvins that is (5Z,7E,9E,13Z,15E,19Z)-docosahexaenoic acid carrying three hydroxy substituents at positions 4, 11 and 17 (the 4S,l lR,17S-stereoisomer). It has a role as an anti-inflammatory agent and a human xenobiotic metabolite. It is a resolvin, a secondary allylic alcohol, a triol and a hydroxy polyunsaturated fatty acid and CAS number: 916888-47- 6.
  • resolvin D5 compound refers to one of the potent proresolving lipid mediators derived from docosahexaenoic acid (DHA). It is a member of the class of resolvins that is (5Z,7S,8E,10Z,13Z,15E,17S,19Z)-7,17-dihydroxydocosa-5,8,10,13,15,19- hexaenoic add . RvD5 has the following structure and CAS number: 578008-43-2.
  • lipoxin B4 compound or “lipoxin B4" or “LXB4” refers to an lipoxygenase-derived eicosanoids formed from arachidonic acid by transcellular metabolism during cell-cell interactions.
  • Lipoxin B4 (LXB4) is produced by the metabolism of 15-HETE or 15(S)-HpETE, and it is a structurally distinct member of the lipoxin family that signals in a manner distinct from LXA4.
  • LXB4 regulates neutrophil activation and mediates antiinflammatory and pro-resolving actions for allergic airway responses.
  • LXB4 has the following structure and CAS number: 98049-69-5:
  • these SPMs are able to have anti-inflammatory effects which are not immunosuppressive. More particularly, these SPMs are able to repair the lesion of airway cell epithelial and to regulate tight junction formation and prevent its disruption during Aspergillus fumigatus infection of CF airway epithelial cells.
  • airway epithelial cells has its general meaning in the art and refers to a continuous layer of epithelial cells that play a critical role in maintaining the conduit for air, and which are central to the defences of the lung against inhaled environmental concomitants. Pathological remodelling of the airway epithelium is commonly observed in CF.
  • the inventors have shown that the SPMs of the invention stimulate cell proliferation and airway epithelial cell repair in order to maintain lung functions in CF.
  • the invention refers to a method for repairing airway epithelial cells in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the invention refers to a method for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the invention refers to a method for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the Aspergillus fumigatus infection is a lung infection.
  • 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 SPM compounds is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • the at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4) and resolvin D5 compound (RvD5) and combinations thereof.
  • 1, 2, 3, 4, 5, 6, 7, 8, or 9 SPM compound is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
  • protectin DI compound PD1
  • RvEl resolvin El compound
  • RvE2 compound resolvin E2 compound
  • RvE3 compound RvE3 compound
  • resolvin E4 compound RvE4 compound
  • 1, 2, 3, 4, 5, 6, 7, 8, or 9 SPM compound selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5) are administered in combination with resolvin DI compound (RvDl), lipoxin B4 (LXB4) compound and/or lipoxin A4 (LXA4) compound.
  • the at least one SPMs is resolvin El compound (RvEl), and/or resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
  • the at least one SPMs is resolvin El compound (RvEl), and/or resolvin D2 compound (RvD2), and/or resolvin D5 compound (RvD5), and/or lipoxin A4 compound (LXA4), and/or lipoxin B4 compound (LXA4) and/or protectin DI compound (PD1).
  • the at least one SPMs are resolvin El compound (RvEl) and lipoxin A4 compound (LXA4).
  • the at least one SPMs are resolvin El compound (RvEl) and/or resolvin D3 compound (RvD3).
  • the at least one SPMs are resolvin El compound (RvEl) and lipoxin B4 compound (LXB4).
  • the at least one SPMs are resolvin E2 compound (RvE2) and/or protectin DI compound (PD1).
  • the at least one SPMs are resolvin El compound (RvEl) and protectin DI compound (PD1).
  • the at least one SPMs are resolvin El compound (RvEl) and/or resolving D3 compound (RvD3).
  • the at least one SPMs are resolvin El compound (RvEl) and/or resolving D5 compound (RvD5).
  • the at least one SPMs are Lipoxin A4 compound (LXA4) and protectin DI compound (PD1).
  • the at least one SPMs are Lipoxin B4 compound (LXB4) and protectin DI compound (PD1).
  • the at least one SPMs are resolving D5 compound (RvD5) and/or protectin DI compound (PD1).
  • the at least one SPMs are Lipoxin A4 compound (LXB4) and lipoxin A4 compound (LXA4).
  • the at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and/or protectin compound (PD1). In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • the at least one SPMs is resolvin El compound (RvEl), lipoxin B4 compound (LXB4), resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
  • the SPMs are resolvin El compound (RvEl), lipoxin B4 compound (LXB4), lipoxin A4 compound (LXA4) resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
  • the at least one SPMs compound is administered in combination with a classical treatment of cystic fibrosis.
  • the at least one SPMs compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4) is administered in combination with a classical treatment of cystic fibrosis.
  • lumacaftor also called “3- ⁇ 6-[l-(2,2-difluoro-2H-l,3- benzodioxol-5-yl)cyclopropaneamido]-3-methylpyridin-2-yl ⁇ benzoic acid” has its general meaning in the art and refers to the compound characterized by the formula of:
  • Orkambi relates to a biotherapy made up of “lumacaftor” and “ivacaftor”. Orkambi is developed by Vertex Pharmaceuticals.
  • Symdeko relates to a biotherapy made up of “tezacaftor” and “ivacaftor”. Symdeko is developed by Vertex Pharmaceuticals.
  • Kaftrio or “Trikfata” relates to a tritherapy made up of “ivacaftor”, “tezacaftor” and “elexacaftor”. Kaftrio/Trikfata is developed by Vertex Pharmaceuticals.
  • Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual, buccal, and inhalation administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms.
  • the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected.
  • Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
  • the at least one SPMs compound according to the invention can be formulated into a composition in a neutral or salt form.
  • the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration.
  • sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure.
  • one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or inj ected at the proposed site of infusion. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
  • the invention refers to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolving El compound (RvEl), resolving E2 compound (RvE2), resolving E3 compound (RvE3), resolving E4 compound (RvE4), resolving DI compound (RvDl), resolving D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the invention refers to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolving El compound (RvEl), resolving E2 compound (RvE2), resolving E3 compound (RvE3), resolving E4 compound (RvE4), resolving D2 compound (RvD2), resolving D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the pharmaceutical composition comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 SPM compounds selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM compounds selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl
  • the pharmaceutical composition comprises 1, 2, 3, 4, 5, 6, 7, 8 or 9 SPM compound is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5) and combinations thereof.
  • protectin DI compound PD1
  • RvEl resolvin El compound
  • RvE2 compound resolvin E2 compound
  • RvE3 compound RvE3 compound
  • resolvin E4 compound RvE4 compound
  • the pharmaceutical composition comprises resolvin El compound (RvEl) and lipoxin A4 compound (LXA4).
  • the pharmaceutical composition comprises resolvin E2 compound (RvE2) and/or protectin compound (PD1).
  • the pharmaceutical composition comprises resolvin El compound (RvEl) and/or protectin compound (PD1). In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl) and/or resolvin D3 compound (RvD3).
  • the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl) and/or resolvin D5 compound (RvD5).
  • the pharmaceutical composition comprises lipoxin A4 compound (LXB4) and lipoxin A4 compound (LXA4).
  • the pharmaceutical composition comprises resolvin El compound (RvEl), lipoxin B4 compound (LXB4) and/or protectin DI compound (PD1).
  • the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and protectin DI compound (PD1).
  • SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and protectin DI compound (PD1).
  • the pharmaceutical composition comprises at least one SPMs are resolvin DI compound (RvDl), resolvin D3 compound (RvD3) and resolvin El compound (RvEl).
  • the pharmaceutical composition comprises at least one SPMs are resolvin DI compound (RvDl), resolvin D5 compound (RvD5) and resolvin El compound (RvEl).
  • the pharmaceutical composition comprises at least one SPMs are protectin DI compound (PD1), and/or resolvin D5 compound (RvD5) and/or resolvin El compound (RvEl).
  • SPMs are protectin DI compound (PD1), and/or resolvin D5 compound (RvD5) and/or resolvin El compound (RvEl).
  • the invention refers to the pharmaceutical composition of the invention for use for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis.
  • the invention refers to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis
  • the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the invention refers to the pharmaceutical composition of the invention for use for treating cystic fibrosis in a patient in need thereof.
  • the invention refers to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for treating cystic fibrosis in a patient in need thereof, wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • the invention refers to the pharmaceutical composition of the invention for use for repairing airway epithelial cell in patient suffering from cystic fibrosis.
  • the invention also refers to a pharmaceutical composition
  • a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for repairing airway epithelial cell in patient suffering from cystic fibrosis, wherein the at least one SPM compound is selected from the group consisting of r protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
  • SPM specialized lipid mediator of the resolution of inflammation
  • FIGURES are a diagrammatic representation of FIGURES.
  • FIG. 1 Airway epithelial cells confluency over time.
  • CFBE41O- cells were plated at 10 000 cells/well on a 96 wells plate and grown in EMEM culture medium without SVF for their synchronization at 37°C and 5% CO2 for 24h. Cells were then treated with SPMs (LXA4, LXB4, RvD2, PD1, RvEl and RvE2) or control solvent (CT EtOH). Images were captured every 2 hours for 84 hours and analyzed using the Incucyte technology.
  • SPMs LXA4, LXB4, RvD2, PD1, RvEl and RvE2
  • CT EtOH control solvent
  • Figure 2 Cell differentiation over time. TEER measurement of CFBE41O- cells (F508del CFTR) grown on permeable filter was followed during liquid/liquid (day 0 to day 20) and air/liquid) interface culture (from day 20). Cells were treated or not with LXA4, LXB4 or RvEl (lOnM) in the basolateral culture compartment the day of plating and the SPM treatment was renewed every second day.
  • LXA4, LXB4 or RvEl LOnM
  • Figure 3 Confocal images of ZO-1 (white) in CFBE41o- cell line. Asperigillus fumigatus (Af) exposure significantly disrupted tight junction organization compared to control condition. LXA4, LXB4, RvEl, PD1 and RvD5 treatment simultaneously with Af exposure prevented ZO-1 disruption.
  • Figure 5 Implication of different receptors in the effect of RvEl, PD1 and Rvd5 on the tight junction organization.
  • Chemerin inhibited the effect of RvEl in preventing tight junction disruption during 4/' exposure, suggesting the competitive effect of chemerin and RvEl on chemR23 receptor.
  • Txl4A inhibited the effect of PD1, suggesting the role of GPR37 receptor in PD1 effect on tight junction protection.
  • the AntiGPR32 inhibited the effect of RvD5, suggesting a role for GPR32 receptor in the effect of RvD5 on tight junction.
  • FIG. 6 SPMs reduce the Asperigillus fumigatus growth in human nasal epithelial cell (hNEC).
  • the Af growth was significantly reduced when Af conidies were inoculated on hNEC (compared with its growth without epithelial cells).
  • PD1 and LXA4 significantly enhanced this effect
  • the CFBE41o- bronchial epithelial cell line over-expressing F508del CFTR mutation or human nasal epithelial cell primary cultures from patients with CF were cultured under airliquid interface to obtain a differentiated epithelium.
  • Af expressing were inoculated at a MOI of 1:6 on the apical side of the epithelium with or without SPMs (lOnM) treatment.
  • the tight junction protein (ZO-1) was stained by immunofluorescence.
  • the Airway surface liquid layer (ASL) was stained using Texas red. Confocal microscopy was used to visualize 4/locati on and tight junction integrity and ASL.
  • TEER Trans Epithelial Electrical Resistance

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Epidemiology (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Pulmonology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The present invention relates to the treatment of fibrosis cystic. The inventors have demonstrated that specialised pro-resolution lipid mediators (SPMs) regulate tight junction formation and prevent its disruption during Aspergillus fumigatus infection of CF airway epithelial cells. Moreover, they have demonstrated that these SPMs repair the CF airway epithelial cells. Thus, the invention relates to methods and pharmaceutical compositions for the treatment of cystic fibrosis. The present invention also relates to methods and pharmaceutical compositions for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis.

Description

SPECIALISED PRO-RESOLUTION LIPID MEDIATOR (SPM) COMPOUNDS FOR USE IN THE TREATMENT OF CYSTIC FIBROSIS AND ASPERGILLUS FUMIGATUS INFECTION IN PATIENT SUFFERING FROM CYSTIC FIBROSIS
FIELD OF THE INVENTION:
The invention is in the field of pneumonology. More particularly, the invention relates to methods and compositions for treating cystic fibrosis airway disease, and in particular repairing/prevent airway epithelial damage. The invention relates also to methods for treating Aspergillus fumigatus infection in patients suffering from cystic fibrosis.
BACKGROUND OF THE INVENTION:
Cystic fibrosis (CF) is an autosomal recessive genetic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene (1). More than 2,100 different mutations have been described affecting protein production or activity. The CFTR chloride channel, mainly expressed in epithelia, regulates transepithelial salt and water fluxes. In CF, impaired chloride ion efflux coupled with sodium ion hyperabsorption results in epithelial surface dehydration and thick and tenacious mucus that obstruct the lungs, pancreatic ducts, the biliary tract, the intestine, and the male reproductive tract. Chronic lung infections are CF’s most common clinical manifestation, leading to pulmonary sustained inflammation, progressive tissue destruction and loss of function, and elevated morbidity and mortality.
Following the cloning of the CFTR gene in 1989 (2), early hope for a therapy to treat patients with CF was founded firmly in the realm of gene therapy (3). For the last 10 years, Vertex Pharmaceuticals’ therapies have revolutionized the clinical approach by providing strategies to address and activate CFTR at the membrane (4). Unfortunately, these approaches are mutation-dependent and more than 15% of patients with nonsense and rare mutations non- responsive to the drugs. Hence the need for alternative therapies that are CFTR independent.
Cystic fibrosis (CF) is characterized by a chronic microbial infection of the respiratory tree associated with persistent inflammation. Aspergillus fumigatus is the predominant filamentous fungus isolated from respiratory cultures of patients with CF and whose prevalence appears to be increasing. Aspergillus is associated with progressive structural lung injury and/or impaired lung function. Aspergillus fumigatus poses a serious health threat to humans by causing numerous invasive infections and a notable mortality in humans, especially in immunocompromised patient. Aspergillus fumigatus is thus included in the critical group (i.e pathogens ranked for perceived public health importance) on the fungal priority pathogens list developed by World health organization.
The inflammatory process is normally self-regulated by an active resolution phase, involving specialized lipid mediators of the resolution of inflammation (SPMs) such as lipoxins (LX), resolvins (Rv), protecins (Pd) and maresins (MaR). Several reports provide evidence for a correlation between chronic inflammatory disease and abnormal production or activity of the specialised pro-resolution lipid mediators (SPMs) including resolvins and lipoxins (5). Previous reports have shown that several SPMs are abnormally produced in CF (6, 7, 8) suggesting that the altered resolution of inflammation in CF is due to a reduced SPMs biosynthesis in the airway of patients. Furthermore, there is a significant correlation between the levels of RvDl in plasma and sputum of CF patients with the biomarkers of inflammation (IL8 and ILip) and lung function (9).
One of the challenges of treating CF airway disease has been to design a therapy which will resolve inflammation, overcome airway epithelial damages and/or will prevent Aspergillus fumigatus infection. Current therapies involve drugs which correct the function of CFTR but this strategy is expensive and do not correct all CFTR mutations. Other strategies have been plagued by the side effects of pro-inflammatory responses.
SUMMARY OF THE INVENTION:
The present invention relates to methods and pharmaceutical compositions for the treatment of cystic fibrosis. The present invention also relates to methods and pharmaceutical compositions for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis. In particular, the present invention is defined by the claims.
DETAILED DESCRIPTION OF THE INVENTION:
The inventors have demonstrated that specialised pro-resolution lipid mediators (SPMs) regulate tight junction formation and prevent its disruption during Aspergillus fumigatus infection of CF airway epithelial cells. They also shown that these SPMs stimulate the epithelial cell immune system and reduce Aspergillus fumigatus growth. Moreover, they have demonstrated that these SPMs repair the CF airway epithelium.
Thus, in a first aspect, the invention relates to a method for treating cystic fibrosis in patient in need thereof comprising administering to said patient a therapeutically effective amount of at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound RvD5, lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In other words, the invention relates to at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound RvD5, lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4), for use for treating cystic fibrosis in patient in need thereof
In particular embodiments, the invention relates to at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound RvD5 for use for treating cystic fibrosis in patient in need thereof
As used herein, the terms “subject” or “patient” denote a mammal, such as a rodent, a feline, a canine, pig, ferret, rat, mice, rabbit and a primate. A subject according to the invention refers to any subject (preferably human) afflicted or at risk to be afflicted with cystic fibrosis. The method of the invention may be performed for any type of cystic fibrosis revised in the World Health Organization Classification of cystic fibrosis and selected from the E84 group: mucoviscidosis or Cystic fibrosis with pulmonary manifestations. In some embodiments, the subject according to the invention is a human. In some embodiments, the subject according to the invention is a girl or a boy. In some embodiments, the subject according to the invention is an adult. In some embodiments, the subject according to the invention is a child (human being between the stages of birth and puberty), a teenager (human being between the stages of puberty to adulthood) or an elderly person (human being after the puberty).
In some embodiment, the patient has or is susceptible to have Aspergillus fumigatus infection. As used herein, the term “Cystic fibrosis” (CF) is a genetic disorder that affects mostly the lungs, but also the pancreas, liver, kidneys, reproductive tract and intestine. Long-term issues include difficulty breathing and coughing up mucus as a result of frequent lung infections and inflammation. Other signs and symptoms may include sinus infections, poor growth, fatty stool, clubbing of the fingers and toes, and infertility in most males. Different people may have different degrees of symptoms.
CF is inherited in an autosomal recessive manner. It is caused by the presence of mutations in both copies of the gene for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Those with a single working copy are carriers and otherwise mostly healthy. CFTR is involved in the production of sweat, digestive fluids, and mucus. When the CFTR is not functional, secretions that are usually thin instead become thick. The condition is diagnosed by a sweat test and genetic testing. Screening of infants at birth takes place in some areas of the world.
As used herein, the term “CFTR protein” refers to the CFTR protein of 1480 amino acids, also called Cystic Fibrosis Transmembrane conductance Regulator. The CFTR protein is a chloride (C1-) channel found in the membranes of secretory tissues as intestinal and respiratory mucosa. The CFTR protein is represented by the NCBI reference sequence: P13569.3 (SEQ: ID NO: 1)
SEQ: ID NO: 1 :
1 mqrsplekas vvsklffswt rpilrkgyrq rlelsdiyqi psvdsadnls eklerewdre
61 laskknpkli nalrrcffwr fmfygiflyl gevtkavqpl llgriiasyd pdnkeersia
121 iylgiglcll fivrtlllhp aifglhhigm qmriamfsli ykktlklssr vldkisigql
181 vsllsnnlnk fdeglalahf vwiaplqval Imgliwellq asafcglgfl ivlalfqagl
241 grmmmkyrdq ragkiserlv itsemieniq svkaycweea mekmienlrq telkltrkaa
301 yvryfhssaf ffsgffvvfl svlpyalikg iilrkiftti sfcivlrmav trqfpwavqt
361 wydslgaink iqdflqkqey ktleynlttt evvmenvtaf weegfgelfe kakqnnnnrk
421 tsngddslff snfsllgtpv Ikdinfkier gqllavagst gagktsllmv imgelepseg
481 kikhsgrisf csqfswimpg tikeniifgv sydeyryrsv ikacqleedi skfaekdniv
541 Igeggitlsg gqrarislar avykdadlyl Idspfgyldv Itekeifesc vcklmanktr 601 ilvtskmehl kkadkililh egssyfygtf selqnlqpdf ssklmgcdsf dqfsaerms 661 iltetlhrfs legdapvswt etkkqsfkqt gefgekrkns ilnpinsirk fsivqktplq 721 mngieedsde plerrlslvp dseqgeailp risvistgpt Iqarrrqsvl nlmthsvnqg 781 qnihrkttas trkvslapqa nlteldiysr rlsqetglei seeineedlk ecffddmesi 841 pavttwntyl ryitvhksli fvliwclvif laevaaslvv Iwllgntplq dkgnsthsm 901 nsyaviitst ssyyvfyiyv gvadtllamg ffrglplvht litvskilhh kmlhsvlqap
961 mstlntlkag gilnrfskdi ailddllplt ifdfiqllli vigaiavvav Iqpyifvatv
1021 pvivafimlr ayflqtsqql kqlesegrsp ifthlvtslk glwtlrafgr qpyfetlfhk 1081 alnlhtanwf lylstlrwfq mriemifvif fiavtfisil ttgegegrvg iiltlamnim 1141 stlqwavnss idvdslmrsv srvfkfidmp tegkptkstk pykngqlskv miienshvkk 1201 ddiwpsggqm tvkdltakyt eggnaileni sfsispgqrv gllgrtgsgk stllsaflrl 1261 Integeiqid gvswdsitlq qwrkafgvip qkvfifsgtf rknldpyeqw sdqeiwkvad 1321 evglrsvieq fpgkldfvlv dggcvlshgh kqlmclarsv Iskakillld epsahldpvt 1381 yqiirrtlkq afadctvilc ehrieamlec qqflvieenk vrqydsiqkl Inerslfrqa 1441 ispsdrvklf phmsskcks kpqiaalkee teeevqdtrl
As used herein, the term “CFTR gene” refers to the CFTR gene which is located on chromosome 7 and which may be found in NCBI GenBank locus AC000111 and AC000061, the contents of which are incorporated herein in their entirety by reference. The cDNA for the CFTR gene is found in Audrezet et al., Hum. Mutat. (2004) 23 (4), 343-357. A nucleic acid sequence for human CFTR is represented by SEQ ID NO: 2.
SEQ ID NO: 2 (NCBI Reference Sequence: NM_000492.3)
1 aattggaagc aaatgacatc acagcaggtc agagaaaaag ggttgagcgg caggcaccca
61 gagtagtagg tctttggcat taggagcttg agcccagacg gccctagcag ggaccccagc
121 gcccgagaga ccatgcagag gtcgcctctg gaaaaggcca gcgttgtctc caaacttttt
181 ttcagctgga ccagaccaat tttgaggaaa ggatacagac agcgcctgga attgtcagac
241 atataccaaa tcccttctgt tgattctgct gacaatctat ctgaaaaatt ggaaagagaa
301 tgggatagag agctggcttc aaagaaaaat cctaaactca ttaatgccct tcggcgatgt
361 tttttctgga gatttatgtt ctatggaatc tttttatatt taggggaagt caccaaagca
421 gtacagcctc tcttactggg aagaatcata gcttcctatg acccggataa caaggaggaa
481 cgctctatcg cgatttatct aggcataggc ttatgccttc tctttattgt gaggacactg
541 ctcctacacc cagccatttt tggccttcat cacattggaa tgcagatgag aatagctatg
601 tttagtttga tttataagaa gactttaaag ctgtcaagcc gtgttctaga taaaataagt
661 attggacaac ttgttagtct cctttccaac aacctgaaca aatttgatga aggacttgca
721 ttggcacatt tcgtgtggat cgctcctttg caagtggcac tcctcatggg gctaatctgg
781 gagttgttac aggcgtctgc cttctgtgga cttggtttcc tgatagtcct tgcccttttt
841 caggctgggc tagggagaat gatgatgaag tacagagatc agagagctgg gaagatcagt
901 gaaagacttg tgattacctc agaaatgatt gaaaatatcc aatctgttaa ggcatactgc
961 tgggaagaag caatggaaaa aatgattgaa aacttaagac aaacagaact gaaactgact
1021 cggaaggcag cctatgtgag atacttcaat agctcagcct tcttcttctc agggttcttt 1081 gtggtgtttt tatctgtgct tccctatgca ctaatcaaag gaatcatcct ccggaaaata
1141 ttcaccacca tctcattctg cattgttctg cgcatggcgg tcactcggca atttccctgg
1201 gctgtacaaa catggtatga ctctcttgga gcaataaaca aaatacagga tttcttacaa
1261 aagcaagaat ataagacatt ggaatataac ttaacgacta cagaagtagt gatggagaat
1321 gtaacagcct tctgggagga gggatttggg gaattatttg agaaagcaaa acaaaacaat
1381 aacaatagaa aaacttctaa tggtgatgac agcctcttct tcagtaattt ctcacttct
1441 ggtactcctg tcctgaaaga tattaatttc aagatagaaa gaggacagtt gttggcggtt
1501 gctggatcca ctggagcagg caagacttca cttctaatgg tgattatggg agaactggag
1561 ccttcagagg gtaaaattaa gcacagtgga agaatttcat tctgttctca gttttcctgg
1621 attatgcctg gcaccattaa agaaaatatc atctttggtg tttcctatga tgaatataga
1681 tacagaagcg tcatcaaagc atgccaacta gaagaggaca tctccaagtt tgcagagaaa
1741 gacaatatag tcttggaga aggtggaatc acactgagtg gaggtcaacg agcaagaatt
1801 tctttagcaa gagcagtata caaagatgct gatttgtatt tattagactc tcctttgga
1861 tacctagatg ttttaacaga aaaagaaata tttgaaagct gtgtctgtaa actgatggct
1921 aacaaaacta ggattttggt cacttctaaa atggaacatt taaagaaagc tgacaaaata
1981 ttaattttgc atgaaggtag cagctatttt tatgggacat tttcagaact ccaaaatcta
2041 cagccagact ttagctcaaa actcatggga tgtgattctt tcgaccaatt tagtgcagaa
2101 agaagaaatt caatcctaac tgagacctta caccgtttct cattagaagg agatgctcct
2161 gtctcctgga cagaaacaaa aaaacaatct tttaaacaga ctggagagtt tggggaaaaa
2221 aggaagaatt ctattctcaa tccaatcaac tctatacgaa aattttccat tgtgcaaaag
2281 actcccttac aaatgaatgg catcgaagag gattctgatg agcctttaga gagaaggctg
2341 tccttagtac cagattctga gcagggagag gcgatactgc ctcgcatcag cgtgatcagc
2401 actggcccca cgcttcaggc acgaaggagg cagtctgtcc tgaacctgat gacacactca
2461 gttaaccaag gtcagaacat tcaccgaaag acaacagcat ccacacgaaa agtgtcactg
2521 gcccctcagg caaacttgac tgaactggat atatattcaa gaaggttatc tcaagaaact
2581 ggcttggaaa taagtgaaga aataacgaa gaagacttaa aggagtgctt ttttgatgat
2641 atggagagca taccagcagt gactacatgg aacacatacc ttcgatatat tactgtccac
2701 aagagcttaa ttttgtgct aatttggtgc ttagtaattt ttctggcaga ggtggctgct
2761 tctttggttg tgctgtggct ccttggaaac actcctcttc aagacaaagg gaatagtact
2821 catagtagaa ataacagcta tgcagtgatt atcaccagca ccagttcgta ttatgtgttt
2881 tacatttacg tgggagtagc cgacactttg cttgctatgg gattcttcag aggtctacca
2941 ctggtgcata ctctaatcac agtgtcgaaa attttacacc acaaaatgtt acatctgtt
3001 cttcaagcac ctatgtcaac cctcaacacg ttgaaagcag gtgggattct taatagattc
3061 tccaaagata tagcaatttt ggatgacctt ctgcctctta ccatattga cttcatccag 3121 ttgttattaa tgtgattgg agctatagca gttgtcgcag ttttacaacc ctacatctt
3181 gttgcaacag tgccagtgat agtggcttt atatgttga gagcatattt cctccaaacc
3241 tcacagcaac tcaaacaact ggaatctgaa ggcaggagtc caattcac tcatctgtt
3301 acaagcttaa aaggactatg gacacttcgt gcctcggac ggcagcctta ctttgaaact
3361 ctgttccaca aagctctgaa tttacatact gccaactggt tcttgtacct gtcaacactg
3421 cgctggttcc aaatgagaat agaaatgatt tttgtcatct tcttcattgc tgtaccttc
3481 atttccattt taacaacagg agaaggagaa ggaagagttg gtattatcct gactttagcc
3541 atgaatatca tgagtacatt gcagtgggct gtaaactcca gcatagatgt ggatagcttg
3601 atgcgatctg tgagccgagt ctttaagttc atgacatgc caacagaagg taaacctacc
3661 aagtcaacca aaccatacaa gaatggccaa ctctcgaaag tatgattat tgagaattca
3721 cacgtgaaga aagatgacat ctggccctca gggggccaaa tgactgtcaa agatctcaca
3781 gcaaaataca cagaaggtgg aaatgccata ttagagaaca tttccttctc aataagtcct
3841 ggccagaggg tgggcctctt gggaagaact ggatcaggga agagtacttt gttatcagct
3901 tttttgagac tactgaacac tgaaggagaa atccagatcg atggtgtgtc ttgggattca
3961 ataactttgc aacagtggag gaaagccttt ggagtgatac cacagaaagt atttatttt
4021 tctggaacat ttagaaaaaa cttggatccc tatgaacagt ggagtgatca agaaatatgg
4081 aaagttgcag atgaggttgg gctcagatct gtgatagaac agtttcctgg gaagcttgac
4141 tttgtccttg tggatggggg ctgtgtccta agccatggcc acaagcagtt gatgtgcttg
4201 gctagatctg ttctcagtaa ggcgaagatc ttgctgcttg atgaacccag tgctcatttg
4261 gatccagtaa cataccaaat aattagaaga actctaaaac aagcatttgc tgattgcaca
4321 gtaattctct gtgaacacag gatagaagca atgctggaat gccaacaatt tttggtcata
4381 gaagagaaca aagtgcggca gtacgattcc atccagaaac tgctgaacga gaggagcctc
4441 tccggcaag ccatcagccc ctccgacagg gtgaagctct ttccccaccg gaactcaagc
4501 aagtgcaagt ctaagcccca gattgctgct ctgaaagagg agacagaaga agaggtgcaa
4561 gatacaaggc tttagagagc agcataaatg ttgacatggg acatttgctc atggaattgg
4621 agctcgtggg acagtcacct catggaattg gagctcgtgg aacagttacc tctgcctcag
4681 aaaacaagga tgaattaagt ttttttttaa aaaagaaaca tttggtaagg ggaattgagg
4741 acactgatat gggtcttgat aaatggcttc ctggcaatag tcaaattgtg tgaaaggtac
4801 ttcaaatcct tgaagattta ccacttgtgt tttgcaagcc agattttcct gaaaaccctt
4861 gccatgtgct agtaattgga aaggcagctc taaatgtcaa tcagcctagt tgatcagctt
4921 attgtctagt gaaactcgtt aatttgtagt gttggagaag aactgaaatc atacttctta
4981 gggttatgat taagtaatga taactggaaa cttcagcggt ttatataagc ttgtattcct
5041 ttttctctcc tctccccatg atgtttagaa acacaactat attgtttgct aagcattcca
5101 actatctcat ttccaagcaa gtattagaat accacaggaa ccacaagact gcacatcaaa 5161 atatgcccca ttcaacatct agtgagcagt caggaaagag aacttccaga tcctggaaat
5221 cagggttagt attgtccagg tctaccaaaa atctcaatat tcagataat cacaatacat
5281 cccttacctg ggaaagggct gtataatct ttcacagggg acaggatggt tcccttgatg
5341 aagaagttga tatgcctttt cccaactcca gaaagtgaca agctcacaga cctttgaact
5401 agagtttagc tggaaaagta tgttagtgca aattgtcaca ggacagccct tctttccaca
5461 gaagctccag gtagagggtg tgtaagtaga taggccatgg gcactgtggg tagacacaca
5521 tgaagtccaa gcatttagat gtataggttg atggtggtat gttttcaggc tagatgtatg
5581 tacttcatgc tgtctacact aagagagaat gagagacaca ctgaagaagc accaatcatg
5641 aattagtttt atatgcttct gttttataat tttgtgaagc aaaatttttt ctctaggaaa
5701 tatttatttt aataatgttt caaacatata taacaatgct gtattttaaa agaatgatta
5761 tgaattacat ttgtataaaa taatttttat atttgaaata ttgacttttt atggcactag
5821 tatttctatg aaatattatg ttaaaactgg gacaggggag aacctagggt gatattaacc
5881 aggggccatg aatcaccttt tggtctggag ggaagccttg gggctgatgc agttgttgcc
5941 cacagctgta tgattcccag ccagcacagc ctcttagatg cagttctgaa gaagatggta
6001 ccaccagtct gactgtttcc atcaagggta cactgccttc tcaactccaa actgactctt
6061 aagaagactg catatattt attactgtaa gaaaatatca cttgtcaata aaatccatac
6121 atttgtgtga aa.
As used herein, the term "gene" has its general meaning in the art and refers to means a DNA sequence that codes for or corresponds to a particular sequence of amino acids which comprise all or part of one or more proteins or enzymes, and may or may not include regulatory DNA sequences, such as promoter sequences, which determine for example the conditions under which the gene is expressed.
As used herein, the “allele" has its general meaning in the art and refers to an alternative form of a gene (one member of a pair) that is located at a specific position on a specific chromosome which, when translated result in functional or dysfunctional (including nonexistent) gene products.
As used herein, the term “mutation” has its general meaning in the art and refers to any detectable change in genetic material, e.g., DNA, RNA, cDNA, or any process, mechanism, or result of such a change. This includes gene mutations, in which the structure (e.g., DNA sequence) of a gene is altered, any gene or DNA arising from any mutation process, and any expression product (e.g., protein or enzyme) expressed by a modified gene or DNA sequence. Mutations include deletion, insertion or substitution of one or more nucleotides. The mutation may occur in the coding region of a gene (i.e., in exons), in introns, or in the regulatory regions (e.g., enhancers, response elements, suppressors, signal sequences, polyadenylation sequences, promoters) of the gene. Generally, a mutation is identified in a subject by comparing the sequence of a nucleic acid or polypeptide expressed by said subject with the corresponding nucleic acid or polypeptide expressed in a control population. Where the mutation is within the gene coding sequence, the mutation may be a “missense” mutation, where it replaces one amino acid with another in the gene product, or a “non-sense” mutation , where it replaces an amino acid codon with a stop codon. A mutation may also occur in a splicing site where it creates or destroys signals for exon-intron splicing and thereby lead to a gene product of altered structure. A mutation in the genetic material may also be “silent”, i.e., the mutation does not result in an alteration of the amino acid sequence of the expression product. In the context of the instant application, mutations identified in CFTR gene or protein are designated pursuant to the nomenclature of Dunnen and Antonarakis (2000). For instance, ">" indicates a substitution at DNA level; (underscore) indicates a range of affected residues, separating the first and last residue affected; "del" indicates a deletion, "dup" indicates a duplication; "ins" indicates a insertion, "inv" indicates an inversion and "con" indicates a conversion. More particularly, “X” denotes that an amino acid is changed to a stop codon (X).
As used herein, the term “homozygous” refers to an individual possessing two copies of the same allele. As used herein, the term “homozygous mutant” refers to an individual possessing two copies of the same allele, such allele being characterized as the mutant form of a gene.
As used herein, the term “heterozygous” refers to an individual possessing two different alleles of the same gene, i.e., an individual possessing two different copies of an allele, such alleles are characterized as mutant forms of a gene.
In some embodiments, the subject harbors at least one mutation in the CFTR gene. The CFTR gene mutations were classified into six classes according to their resulting damaging effect on the protein (Elbom JS, 2016). The class I mutations contribute to the formation of proteins with incomplete length and usually involve the complete loss of its activity (e.g.,W1282X, 1717-1G->A, G542X, R553X, 2183AA>G). Mutation in the class II lead to abnormal maturation of proteins in the endoplasmic reticulum and Golgi apparatus. The effect of these mutations is premature degradation of the CFTR protein. Hence, CFTR protein does not reach the cell membrane where it should perform its function (e.g., F508del, 2184delA). The gene product having mutations of class III is properly synthesized, transported, and incorporated into the cell membrane, but has decreased activity caused by abnormal regulation of the protein. These mutations are frequently situated within one of the nucleotide binding domains, (e.g., G551D, R560T). Mutations of class IV cause anomalies in the structure of the transmembrane protein and thereby reduce the conduction of the chloride channel (e.g., R117H, R334W). Mutations altering the stability of the mRNA coding for the CFTR protein represent a class V of the mutations of the CFTR gene (e.g., 2789+5G->A, A455E). Mutations in class VI lead to a decrease of the CFTR protein stability (e.g., 4326delTC, Glnl412X, 4279insA) (Harriet C., Kristin E. T., Olivier T., et al. Translating the genetics of cystic fibrosis to personalized medicine. Transl Res 2016; 168 40-49 and Elbom JS. Cystic fibrosis. Seminar. 2016).
Examples of CFTR mutations include, but are not limited to 124del23bp CFTR, CFTRdelel CFTR, M1V CFTR, Q2X CFT, S4X CFTR, P5L CFTR, S13F CFTR, L15P CFTR, 182delT CFTR, CFTRdele2 CFTR, CFTRdele2-4 CFTR, 185+1G->T CFTR, CFTRdele2,3 CFTR, W19X CFTR, G27R CFTR, G27X CFTR, Q30X CFTR, R31C CFTR, R31L CFTR, Q39X CFTR, A46D CFTR, 296+lG->A CFTR, 296+lG->T CFTR, CFTRdele3-10,14b-16 CFTR, 296+28A->G CFTR, 296+2T->C CFTR, 296+3insT CFTR, 297-3C->T CFTR, 297- 1G->A CFTR, E56K CFTR, W57G CFTR, W57X CFTR, 306insA CFTR, 306delTAGA CFTR, E60X CFTR, P67L CFTR, R74W CFTR, R75X CFTR, R75Q CFTR, 365-366insT CFTR, G85E CFTR, 394delTT CFTR, L88X CFTR, G91R CFTR, CFTRdele4-7 CFTR, CFTRdele4-l l CFTR, CFTR50kbdel CFTR, 4O5+1G->A CFTR, 405+3A->C CFTR, 406-2A- >G CFTR, 4O6-1G->A CFTR, E92K CFTR, E92X CFTR, Q98X CFTR, Q98R CFTR, P99L CFTR, L102R CFTR, 442delA CFTR, 444delA CFTR, 457TAT->G CFTR, D110H CFTR, D110E CFTR, R117C CFTR, R117G CFTR, R117H CFTR, R117H;5T CFTR, R117H;7T CFTR, 541 del C CFTR, L138ins CFTR, H139RCFTR, 574delA CFTR, I148T CFTR, 602dell4 CFTR, Y161D CFTR , 621+1G->T CFTR, 621+3A->G CFTR, L165S CFTR, R170H CFTR, 663delT CFTR, G178R CFTR, 675del4 CFTR, E193X CFTR, 711+1G->T CFTR, 711+3A- >G CFTR, 711+5G->A CFTR, 712-1G->T CFTR, H199Y CFTR, V201M CFTR, P205S CFTR, L206W CFTR, W216X CFTR, Q220X CFTR, L227R CFTR, V232D CFTR, 849delG CFTR, 852del22 CFTR, CFTRdup6b-10 CFTR, M265R CFTR, 935delA CFTR, Y275X CFTR, C276X CFTR, 977insA CFTR, 991del5 CFTR, F31 IL CFTR, 1078delT CFTR, L320V CFTR, 1119delA CFTR, G330X CFTR, R334W CFTR, R334Q CFTR, R334L CFTR, 1138insG CFTR, I336K CFTR, T338I CFTR, S341P CFTR, 1154insTC CFTR, 1161delC CFTR, R347H CFTR, R347P CFTR, A349V CFTR, R352W CFTR, R352Q CFTR, Q359K/T360K CFTR, 1213delT CFTR, 1248+1G->A CFTR, 1249-1G->A CFTR, 1259insA CFTR, 1288insTA CFTR, W401X CFTR, 1341+1G->A CFTR, 5T CFTR, 5T;TG11 CFTR, 5T;TG12 CFTR, 5T;TG13 CFTR, 7T CFTR, 9T CFTR, 1343delG CFTR, Q414X CFTR, 1429del7 CFTR, D443Y CFTR, 1461ins4 CFTR, 1471delA CFTR, L453S CFTR, A455E CFTR, 1497delGG CFTR, V456A CFTR, 1504delG CFTR, 1525- 1G->A CFTR, 1525-2A->G CFTR, S466X CFTR, L467P CFTR, M470V CFTR, 1548delG CFTR, E474K CFTR, S489X CFTR, S492F CFTR, 1609delCA CFTR, Q493X CFTR, W496X CFTR, I502T CFTR, I507del CFTR, F508del CFTR, F508C CFTR, D513GCFTR, 1677delTA CFTR, V520F CFTR, C524X CFTR, Q525X CFTR, 1716+1G->A CFTR, CFTRdelel l CFTR, 1717-1G->A CFTR, 1717- 8G->A CFTR, G542X CFTR, S549R CFTR, S549N CFTR, G550X CFTR, 1782delA CFTR, G551S CFTR, G551D CFTR, Q552X CFTR, R553X CFTR, 1802delC CFTR, L558S CFTR, A559T CFTR, 1811+1G->C CFTR, R560K CFTR, R560T CFTR, 1811+1G->A CFTR, 1811+1634A->G or 1811+1.6kbA->G CFTR, 1811+1643G->T CFTR, 1812-1G->A CFTR, R560S CFTR, A561E CFTR, V562I CFTR, Y563N CFTR, Y563D CFTR, 1824delA CFTR, 1833delT CFTR, Y569D CFTR , P574H CFTR , F575Y CFTR, G576A CFTR, D579G CFTR, E585X CFTR, E588V CFTR, 1898+1G->A CFTR, 1898+1G->C CFTR, 1898+1G->T CFTR, CFTRdelel3,14a CFTR, 1898+3A->G CFTR, 1898+5G->T CFTR, 1924del7 CFTR, H609R CFTR, A613T CFTR, D614G CFTR, G622D CFTR , 2055del9->A CFTR, 2075delA CFTR, 2105-2117dell3insAGAAA CFTR, 2118del4 CFTR, R668C CFTR, 2143delT CFTR, G673X CFTR, 2183AA->G CFTR, 2184insA CFTR, 2184delA CFTR, 2185insC CFTR, Q685X CFTR, R709X CFTR, K710X CFTR, Q715X CFTR, Q720X CFTR, 2307insA CFTR, L732X CFTR, 2347delG CFTR, 2372del8 CFTR, P750L CFTR, V754M CFTR, R764X CFTR, R785X CFTR, R792X CFTR, I807M CFTR, 2556insAT CFTR, 2585delT CFTR, 2594delGT CFTR, E822X CFTR, 2622+1 G->A CFTR, E831X CFTR, D836Y CFTR, W846X CFTR Y849X CFTR, R851X CFTR, T854T CFTR, 2711delT CFTR, 2721delll CFTR, 2732insA CFTR, CFTRdelel4b-17b CFTR, 2752-26A->G CFTR, W882X CFTR, 2789+2insA CFTR, 2789+5G->A CFTR, 279O-1G->C CFTR, Q890X CFTR, S912X CFTR, S912L CFTR, 2869insG CFTR, Y913X CFTR, 2896insAG CFTR, L927P CFTR, 2942insT CFTR, 2957delT CFTR, S945L CFTR, 2991del32 CFTR, 3007delG CFTR, 3028delA CFTR, L967S CFTR, G970R CFTR, CFTRdelel6-17b CFTR, G970D CFTR, S977F CFTR, D979V CFTR, 3120G- >A CFTR, CFTRdelel7a,17b CFTR, CFTRdelel7a-18 CFTR, 312O+1G->A CFTR, 3121-1G- >A CFTR, 3121-2A->G CFTR, 3121-977_3499+248del2515 CFTR, L997F CFTR, 3132delTG CFTR, A1006E CFTR, 3143del9 CFTR, 3171delC CFTR, 3171insC CFTR, Y1014C CFTR, F1016S CFTR, I1027T CFTR, Y1032C CFTR, Q1042X CFTR, 3271delGG CFTR, 3272-26A->G CFTR, F1052V CFTR, T1053I CFTR, H1054D CFTR, G1061R CFTR, L1065P CFTR, R1066C CFTR, R1066H CFTR, G1069R CFTR, R1070W CFTR, R1070Q CFTR, 3349insT CFTR, F1074L CFTR, L1077P CFTR, W1089X CFTR, Y1092X CFTR, W1098X CFTR, W1098C CFTR, F1099L CFTR, M1101K CFTR, R1102X CFTR, E1104X CFTR, S1118F CFTR, CFTRdelel8 CFTR, 3500-2A->G CFTR, W1145X CFTR, D1152H CFTR, V1153E CFTR, 3600G->A CFTR, CFTRdelel9 CFTR, CFTRdelel9-21 CFTR, 3600+2insT CFTR, 3600+5G->A CFTR, R1158X CFTR, S1159P CFTR, S1159F CFTR, R1162X CFTR, R1162L CFTR, 3659delC CFTR, 3667ins4 CFTR, S1196X CFTR, 3737delA CFTR, W1204X CFTR, 3791delC CFTR, Y122X CFTR, 3821delT CFTR, I1234V CFTR, S1235R CFTR, 3849G->A CFTR, 3849+4A->G CFTR, 3849+5G->A CFTR, 3849+40A->G CFTR, 3849+10kbC->T CFTR, 385O-1G->A CFTR, 3850-3T->G CFTR, V1240G CFTR, G1244E CFTR, T1246I CFTR, 3876delA CFTR, 3878delG CFTR, S1251N CFTR, L1254X CFTR, S1255P CFTR, S1255X CFTR, 3905insT CFTR, D1270N CFTR, W1282X CFTR, R1283M CFTR, Q1291R CFTR, 4OO5+1G->A CFTR, CFTRdele21 CFTR, 4005+2T->C CFTR, 4010del4 CFTR, 4015delA CFTR, 4016insT CFTR, 4022insT CFTR, 4040delA CFTR, N1303K CFTR, Q1313X CFTR, CFTRdele22-24 CFTR, CFTRdele22,23 CFTR, L1324P CFTR, Q1330X CFTR, L1335P CFTR, 4168delCTAAGCC CFTR, G1349D CFTR, 4209TGTT->AA CFTR, 4218insT CFTR, E1371X CFTR, H1375P CFTR, 4259del5 CFTR, Q1382X CFTR, 4279insA CFTR, 4326delTC CFTR, Q 141 IX CFTR, Q1412X CFTR, 4374+lG->T CFTR, 4374+lG->A CFTR, 4382delA CFTR, 4428insGA CFTR, A1067T CFTR, E193K CFTR, K1060T CFTR, Glnl412X CFTR (see, e g., https://www.cftr2.org/mutations_history, for CFTR mutations).
In one embodiment, the subject harbors at least one allelic mutation selected from class
I, class II, class III, class IV, class V or class VI.
In one embodiment, the subject harbors at least one mutation selected from class I, class
II, class III, class IV, class V or class VI in the first allele and at least one mutation selected from class I, class II, class III, class IV, class V or class VI in the second allele. In a particular embodiment, the subject harbors at least a mutation of class I in the first allele and at least a mutation of class II in the second allele. In a particular embodiment, the subject harbors at least a mutation of class II in the first allele and at least a mutation of class II in the second allele.
In a particular embodiment, the subject harbors at least one allelic mutation in the CFTR gene, including but not limited to F508del-CFTR, R117H CFTR, 2184delA CFTR, W1282X CFTR, 2183AA>G CFTR or G551D CFTR.
In one embodiment, the subject harbors at least a F508del mutation in the CFTR gene.
In one embodiment, the subject harbors at least a 2183AA>G mutation in the CFTR gene. In one embodiment, the subject harbors at least a F508del mutation in the first allele and at least a 2183AA>G mutation in the second allele. In one embodiment, the subject harbors at least a W1282X mutation in the CFTR gene.
As used herein, the term "treatment" or "treat" refer to both prophylactic or preventive treatment as well as curative or disease-modifying treatment, including treatment of subjects at risk of contracting the disease or suspected to have contracted the disease as well as subjects who are ill or have been diagnosed as suffering from a disease or medical condition, and includes suppression of clinical relapse. The treatment may be administered to a subject having a medical disorder or who ultimately may acquire the disorder in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of a disorder or recurring disorder, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment. By "therapeutic regimen" is meant the pattern of treatment of an illness, e.g., the pattern of dosing used during therapy. A therapeutic regimen may include an induction regimen and a maintenance regimen. The phrase "induction regimen" or "induction period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the initial treatment of a disease. The general goal of an induction regimen is to provide a high level of drug to a subject during the initial period of a treatment regimen. An induction regimen may employ (in part or in whole) a "loading regimen", which may include administering a greater dose of the drug than a physician would employ during a maintenance regimen, administering a drug more frequently than a physician would administer the drug during a maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance period" refers to a therapeutic regimen (or the portion of a therapeutic regimen) that is used for the maintenance of a subject during treatment of an illness, e.g., to keep the subject in remission for long periods of time (months or years). A maintenance regimen may employ continuous therapy (e.g., administering a drug at regular intervals, e.g., weekly, monthly, yearly, etc.) or intermittent therapy (e.g., interrupted treatment, intermittent treatment, treatment at relapse, or treatment upon achievement of particular predetermined criteria [e.g., disease manifestation, etc.]).
As used herein the terms "administering" or "administration" refer to the act of injecting or otherwise physically delivering a substance as it exists outside the body (e.g., at least one SPMs of the invention) into the subject, such as by, intravenous, intramuscular, enteral, subcutaneous, parenteral, systemic, local, spinal, nasal, topical or epidermal administration (e.g., by injection or infusion). When a disease, or a symptom thereof, is being treated, administration of the substance typically occurs after the onset of the disease or symptoms thereof. When a disease or symptoms thereof, are being prevented, administration of the substance typically occurs before the onset of the disease or symptoms thereof. According to the invention, the drug is administered to the subject using any suitable method that enables the drug to reach the lungs, pancreas, intestine, skin, reproductive system. In some embodiments, the drug administered to the patients systemically (i.e., via systemic administration). Thus, in some embodiments, the drug is administered to the subject such that it enters the circulatory system and is distributed throughout the body. In some embodiments, the drug is administered to the subject by local administration. In a particular embodiment, the at least one SPMs of the invention is administered by oral administration. In a particular embodiment, the at least one SPMs of the invention is administered by nasal administration. In a particular embodiment, the at least one SPMs of the invention is administered by inhalation.
A “therapeutically effective amount” is intended for a minimal amount of active agent which is necessary to impart therapeutic benefit to a subject. For example, a "therapeutically effective amount" to a subject is such an amount which induces, ameliorates or otherwise causes an improvement in the pathological symptoms, disease progression or physiological conditions associated with or resistance to succumbing to a disorder. It will be understood that the total daily usage of the compounds of the present invention will be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts. For example, it is well within the skill of the art to start doses of the compound at levels lower than those required to achieve the desired therapeutic effect and to gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day. Typically, the compositions contain 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500 mg of the active ingredient for the symptomatic adjustment of the dosage to the subject to be treated. A medicament typically contains from about 0.01 mg to about 500 mg of the active ingredient, preferably from 1 mg to about 100 mg of the active ingredient. An effective amount of the drug is ordinarily supplied at a dosage level from 0.0002 mg/kg to about 20 mg/kg of body weight per day, especially from about 0.001 mg/kg to 7 mg/kg of body weight per day.
As used herein, the terms “specialized lipid mediators of the resolution of inflammation” or “specialized pro-resolving mediators (SPMs)” has its general meaning in the art and refers to a class of cell signaling molecules formed in the organism by the metabolism of polyunsaturated fatty acids (PUFA) by a combination of lipoxygenase, cyclooxygenase, and cytochrome P450 monooxygenase enzymes. SPM are metabolites of arachidonic acid (AA), eicosapentaenoic acid (EP A), docosahexaenoic acid (DHA), or n-3 DPA (i.e. 7,10Z,13Z,19Z-docosapentaenoic acid or clupanodonic acid). The main families of SPM identified include lipoxins (LX), resolvins (Rv), and protectins (PD) and maresins (MaR). They limit inflammation by inhibiting the synthesis and function of pro-inflammatory cytokines as well as leukocyte chemotaxis and migration. SPMs enhance innate microbial killing and clearance by increasing leukocytes’ phagocytosis capacity.
According to the invention, SPMs includes protectin DI (PD1), resolvin El (RvEl), resolvin E2 (RvE2), resolvin E3 (RvE3), resolvin E4 (RvE4), resolvin DI (RvDl), resolvin D2 (RvD2), resolvin D3 (RvD3), resolvin D4 (RvD4), resolvin D5 (RvD5), lipoxin A4 (LXA4) and lipoxin B4 (LXB4).
As used herein, the term “protectin DI compound” , or "‘protectin DI” or “PD1” refers to a specialized pro-resolving mediator (SPM) synthesized from docosahexaenoic acid. Protectin DI increases phagocytosis of apoptotic polymorphonuclear leukocytes (PMNs) by macrophages in a non-phlogistic manner and is generated in vitro during macrophage-apoptotic interactions. PD1 has the following structure and CAS number: 660430-03-5:
Figure imgf000016_0001
As used herein, the term “resolvin El compound” or “resolvin El” or “RvEl” refers to an anti-inflammatory lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid (EP A) that has been shown to promote inflammatory resolution in numerous disease models, including experimental colitis, asthma, atherosclerosis, type 2 diabetes, and HSV-1 -induced stromal keratitis. RvElis biosynthesized by neutrophils via the 5-lipoxygenase pathway.
RvEl has the following structure and CAS number: 552830-51-0:
Figure imgf000017_0001
As used herein, the term “resolvin E2 compound” or “resolvin E2" or “RvE2” refers to an anti-inflammatory lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid (EP A) that has been shown to regulating chemotaxis of human neutrophils; and enhancing phagocytosis and anti-inflammatory cytokine production.
RvE2 has the following structure and CAS number: 865532-70-3
Figure imgf000017_0002
As used herein, the term “resolvin E3 compound” or “resolvin E3” or “RvE3” refers to a specialized pro-resolving lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid (EP A). \ mrsqj %v{J6%Esi%v{j7tfwj ?gitx~synjxn j i?g~fej zy:umKj%{if%znj %: ST ] %ufyn | f~W {J8% ixfi j sjvfyi i%j~%j txrBtumKcp^if%mj ^6746 : 22T ] 3
RvE3 has the following structure:
Figure imgf000017_0003
As used herein, the term “resolvin E4 compound” or “resolvin E4 “ or “RvE4” refers to a proresolving lipid mediator derived from omega-3 fatty acid eicosapentaenoic acid (EP A) that stimulates efferocytosis of apoptotic neutrophils or senescent red blood cells by macrophages, RvE4 is biosynthesized by macrophages or neutrophils via the 5-lipoxygenase and 15 -lipoxygenase pathways. RvE4 has the following structure and CAS number: 1781227-
Figure imgf000018_0001
87-9
As used herein, the term “resolvin DI compound” or “resolvin DI” or “RvDl” refers to one of the potent lipid mediators derived from both eicosapentaenoic acid (EP A) and docosahexaenoic acid (DHA). Resolvin DI (RvDl) is produced physiologically from the sequential oxygenation of DHA by 15- and 5-lipoxygenase. In addition to being antiinflammatory, resolvins promote the resolution of the inflammatory response back to a noninflamed state. RvDl has the following structure and CAS number: 872993-05-0:
Figure imgf000018_0002
As used herein, the term “resolvin D2 compound” or “resolvin D2" or “RvD2” refers to one of the potent lipid mediators derived from docosahexaenoic acid (DHA). Resolvin D2 (RvD2) is produced physiologically from the sequential oxygenation of DHA by 15- and 5- lipoxygenase and functions to dampen excessive neutrophil trafficking to sites of inflammation. RvD2 has the following structure and CAS number: 810668-37-2:
Figure imgf000018_0003
As used herein, the term “resolvin D3 compound” or “resolvin D3” or “RvD3” refers to one of the potent lipid mediators derived from docosahexaenoic acid (DHA). member of the class of resolvins that is (5Z,7E,9E,13Z,15E,19Z)-docosahexaenoic acid carrying three hydroxy substituents at positions 4, 11 and 17 (the 4S,l lR,17S-stereoisomer). It has a role as an anti-inflammatory agent and a human xenobiotic metabolite. It is a resolvin, a secondary allylic alcohol, a triol and a hydroxy polyunsaturated fatty acid and CAS number: 916888-47- 6.
Figure imgf000019_0001
As used herein, the term “resolvin D4 compound” or “resolvin D4” or “RvD4” refers to one of the potent lipid mediators derived from docosahexaenoic acid (DHA). It is a member of the class of resolvins that is (6E,8E,10Z,13Z,15E,19Z)-docosahexaenoic acid carrying three hydroxy substituents at positions 4, 5 and 17 (the 4S,5R,17S-stereoisomer). It has a role as an anti-inflammatory agent and a human xenobi otic metabolite. It is a resolvin, a secondary allylic alcohol, a triol and a hydroxy polyunsaturated fatty acid, and CAS number: 1025684-60-9.
Figure imgf000019_0002
As used herein, the term “resolvin D5 compound” refers to one of the potent proresolving lipid mediators derived from docosahexaenoic acid (DHA). It is a member of the class of resolvins that is (5Z,7S,8E,10Z,13Z,15E,17S,19Z)-7,17-dihydroxydocosa-5,8,10,13,15,19- hexaenoic add . RvD5 has the following structure and CAS number: 578008-43-2.
Figure imgf000020_0001
As used herein, the term “lipoxin A4 compound” or “lipoxin A4 “ or “LXA4” refers to an lipoxygenase-derived eicosanoids formed from arachidonic acid by transcellular metabolism during cell-cell interactions. LXA4 has potent dual pro-resolving and antiinflammatory properties. Lipoxin A4 (LXA4) is produced by the metabolism of 15-HETE or 15(S)-HpETE. LXA4 has the following structure and CAS number: 89663-86-5:
Figure imgf000020_0002
As used herein, the term “lipoxin B4 compound” or “lipoxin B4" or “LXB4” refers to an lipoxygenase-derived eicosanoids formed from arachidonic acid by transcellular metabolism during cell-cell interactions. Lipoxin B4 (LXB4) is produced by the metabolism of 15-HETE or 15(S)-HpETE, and it is a structurally distinct member of the lipoxin family that signals in a manner distinct from LXA4. LXB4 regulates neutrophil activation and mediates antiinflammatory and pro-resolving actions for allergic airway responses. LXB4 has the following structure and CAS number: 98049-69-5:
Figure imgf000020_0003
In the context of the invention, these SPMs are able to have anti-inflammatory effects which are not immunosuppressive. More particularly, these SPMs are able to repair the lesion of airway cell epithelial and to regulate tight junction formation and prevent its disruption during Aspergillus fumigatus infection of CF airway epithelial cells. As used herein, the term “airway epithelial cells” has its general meaning in the art and refers to a continuous layer of epithelial cells that play a critical role in maintaining the conduit for air, and which are central to the defences of the lung against inhaled environmental concomitants. Pathological remodelling of the airway epithelium is commonly observed in CF. Indeed, CF is associated with a severe clinical phenotype that leads to chronic inflammation and infection of the airways by opportunistic pathogens, including bacteria such as Pseudomonas aeruginosa and/or fungi such as, Aspergillus fumigatus. The continuous exposure to severe harmful stimuli places lungs at constant risk of injury and thereby, tissue repair is crucial for maintaining lung homeostasis. CFTR plays a key role in regeneration of the airway epithelium, the repair of which is obviously insufficient to maintain lung functions in CF.
The inventors have shown that the SPMs of the invention stimulate cell proliferation and airway epithelial cell repair in order to maintain lung functions in CF.
Thus, in some embodiments, the at least one SPM repairs airway epithelial cells.
Accordingly, in another aspect, the invention refers to a method for repairing airway epithelial cells in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, the invention refers to a method for repairing airway epithelial cells in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), and resolvin D5 compound (RvD5).
In some embodiment, the patient has or is susceptible to have Aspergillus fumigatus infection. In some embodiment, the patient has lung injury caused by Aspergillus fumigatus infection. Indeed, the inventors show the role of the SPMs of the invention in regulating tight junction formation and preventing its disruption during Aspergillus fumigatus infection of CF airway epithelial cells.
The inventors also show that the SPMs stimulate the epithelial cell immune system and thus reduce the growth of Aspergillus fumigatus.
Accordingly, in another aspect, the invention refers to a method for treating or preventing Aspergillus fumigatus infection in patient in need thereof comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, the patient suffers from cystic fibrosis.
Accordingly, in another aspect, the invention refers to a method for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, the invention refers to a method for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
As used herein, the term “Aspergillus fumigatus infection” or “aspergillosis” has its general meaning in the art and refers to a fungal infection of usually the lungs caused by the genus Aspergillus, a common mold that is breathed in frequently from the air around, but does not usually affect most people. It generally occurs in people with lung diseases such as asthma, cystic fibrosis or tuberculosis. Aspergillus fumigatus is the main fungus cultured in the airways of patients with cystic fibrosis (CF). Allergic bronchopulmonary aspergillosis occurs in -10% of CF patients and is clearly associated with airway damage and lung function decline.
In particular embodiment, the Aspergillus fumigatus infection is a lung infection.
In some embodiments regarding all the methods of the invention, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 SPM compounds is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, regarding all the methods of the invention, the at least one specialized lipid mediators of the resolution of inflammation (SPMs) compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolving E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4) and resolvin D5 compound (RvD5) and combinations thereof.
In some embodiments, regarding all the methods of the invention, 1, 2, 3, 4, 5, 6, 7, 8, or 9 SPM compound is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
In some embodiments, regarding all the methods of the invention, 1, 2, 3, 4, 5, 6, 7, 8, or 9 SPM compound selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5) are administered in combination with resolvin DI compound (RvDl), lipoxin B4 (LXB4) compound and/or lipoxin A4 (LXA4) compound.
In some embodiments, regarding all the methods of the invention, the at least one SPMs is resolvin El compound (RvEl), and/or resolvin D5 compound (RvD5) and/or protectin DI compound (PD1). In some embodiments, regarding the method for treating or preventing Aspergillus fumigatus infection of the invention, the at least one SPMs is resolvin El compound (RvEl), and/or resolvin D2 compound (RvD2), and/or resolvin D5 compound (RvD5), and/or lipoxin A4 compound (LXA4), and/or lipoxin B4 compound (LXA4) and/or protectin DI compound (PD1).
In some embodiments, regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and lipoxin A4 compound (LXA4).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and/or resolvin D3 compound (RvD3).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and lipoxin B4 compound (LXB4).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin E2 compound (RvE2) and/or protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and/or resolving D3 compound (RvD3).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl) and/or resolving D5 compound (RvD5).
In some embodiments regarding all the methods of the invention, the at least one SPMs are Lipoxin A4 compound (LXA4) and protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are Lipoxin B4 compound (LXB4) and protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolving D5 compound (RvD5) and/or protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are Lipoxin A4 compound (LXB4) and lipoxin A4 compound (LXA4).
In some embodiments regarding all the methods of the invention, the at least one SPMs is resolvin El compound (RvEl), lipoxin B4 compound (LXB4) and/or protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and/or protectin compound (PD1). In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin DI compound (RvDl), resolvin D3 compound (RvD3) and resolvin El compound (RvEl).
In some embodiments regarding all the methods of the invention, the at least one SPMs are resolvin DI compound (RvDl), resolvin D5 compound (RvD3) and resolvin El compound (RvEl).
In some embodiments regarding all the methods of the invention, the at least one SPMs are protectin DI compound (PD1), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In some embodiments, regarding all the methods of the invention, the at least one SPMs are resolvin El compound (RvEl), resolvin D5 compound (RvD5) and protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the at least one SPMs is resolvin El compound (RvEl), lipoxin B4 compound (LXB4), resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
In some embodiments regarding all the methods of the invention, the SPMs are resolvin El compound (RvEl), lipoxin B4 compound (LXB4), lipoxin A4 compound (LXA4) resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
In a particular embodiment regarding all the methods of the invention, the at least one SPMs compound is administered in combination with a classical treatment of cystic fibrosis.
In a particular embodiment regarding all the methods of the invention, the at least one SPMs compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4) is administered in combination with a classical treatment of cystic fibrosis.
In a particular embodiment regarding all the methods of the invention, the at least one SPMs compound selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4) resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4) and resolvin D5 compound (RvD5) is administered in combination with a classical treatment of cystic fibrosis.
As used herein, the term “classical treatment of cystic fibrosis” refers to the treatments used or will be used to treat cystic fibrosis airways. Typically, the classical treatment includes, but is not limited to: antibiotics, aerosolized medications (e.g. domase alfa, hypertonic saline, Denufosol), chest physiotherapy, expiratory pressure physiotherapy, anti-inflammatory compounds (Lenabasum also known as Corbus, Celtaxis), Lung transplantation, lumacaftor, ivacaftor or tezacaftor.
As used herein the term “tezacaftor” also called “l-(2,2-difluoro-2H-l,3-benzodioxol- 5-yl)-N-{l-[(2R)-2,3-dihydroxypropyl]-6-fluoro-2-(l-hydroxy-2-methylpropan-2-yl)-lH- indol-5-yl}cyclopropane-l-carboxamide” has its general meaning in the art and refers to the compound characterized by the formula of:
Figure imgf000026_0001
As used herein, the term “lumacaftor” also called “3-{6-[l-(2,2-difluoro-2H-l,3- benzodioxol-5-yl)cyclopropaneamido]-3-methylpyridin-2-yl}benzoic acid” has its general meaning in the art and refers to the compound characterized by the formula of:
Figure imgf000026_0002
As used herein, the term “ivacaftor” also called “N-(2,4-di-tert-butyl-5- hydroxyphenyl)-4-oxo-l,4-dihydroquinoline-3-carboxamide” has its general meaning in the art and refers to the compound characterized by the formula of:
Figure imgf000027_0001
As used herein, the term “elexacaftor” also called “N-(l,3-dimethylpyrazol-4- yl)sulfonyl-6-[3-(3,3,3-trifluoro-2,2-dimethylpropoxy)pyrazol-l-yl]-2-[(4S)-2,2,4- trimethylpyrrolidin-l-yl]pyridine-3 -carboxamide” has its general meaning in the art and refers to the compound characterized by the formula of:
Figure imgf000027_0002
As used herein, the term “Orkambi” relates to a biotherapy made up of “lumacaftor” and “ivacaftor”. Orkambi is developed by Vertex Pharmaceuticals.
As used herein, the term “Symdeko” relates to a biotherapy made up of “tezacaftor” and “ivacaftor”. Symdeko is developed by Vertex Pharmaceuticals.
As used herein, the term “Kaftrio” or “Trikfata” relates to a tritherapy made up of “ivacaftor”, “tezacaftor” and “elexacaftor”. Kaftrio/Trikfata is developed by Vertex Pharmaceuticals.
As used herein, the terms “combined treatment”, “combined therapy” or “therapy combination” refer to a treatment that uses more than one medication. The combined therapy may be dual therapy or bi-therapy.
As used herein, the term “administration simultaneously” refers to administration of 1, 2 or 3 active ingredients by the same route and at the same time or at substantially the same time. The term “administration separately” refers to an administration of 3 active ingredients at the same time or at substantially the same time by different routes. The term “administration sequentially” refers to an administration of 3 active ingredients at different times, the administration route being identical or different. The at least one SPMs compound according to the invention alone and/or with a classical treatment as described above may be combined with pharmaceutically acceptable excipients, and optionally sustained-release matrices, such as biodegradable polymers, to form pharmaceutical compositions. “Pharmaceutically” or “pharmaceutically acceptable” refer to molecular entities and compositions that do not produce an adverse, allergic or other untoward reaction when administered to a mammal, especially a human, as appropriate. A pharmaceutically acceptable carrier or excipient refers to anon-toxic solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type. The pharmaceutical compositions of the present invention for oral, sublingual, subcutaneous, intramuscular, intravenous, transdermal, local or rectal administration, the active principle, alone or in combination with another active principle, can be administered in a unit administration form, as a mixture with conventional pharmaceutical supports, to animals and human beings. Suitable unit administration forms comprise oral-route forms such as tablets, gel capsules, powders, granules and oral suspensions or solutions, sublingual, buccal, and inhalation administration forms, aerosols, implants, subcutaneous, transdermal, topical, intraperitoneal, intramuscular, intravenous, subdermal, transdermal, intrathecal and intranasal administration forms and rectal administration forms. Typically, the pharmaceutical compositions contain vehicles which are pharmaceutically acceptable for a formulation capable of being injected. These may be in particular isotonic, sterile, saline solutions (monosodium or disodium phosphate, sodium, potassium, calcium or magnesium chloride and the like or mixtures of such salts), or dry, especially freeze-dried compositions which upon addition, depending on the case, of sterilized water or physiological saline, permit the constitution of injectable solutions. The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; formulations including sesame oil, peanut oil or aqueous propylene glycol; and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi. Solutions comprising compounds of the invention as free base or pharmacologically acceptable salts can be prepared in water suitably mixed with a surfactant, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The at least one SPMs compound according to the invention can be formulated into a composition in a neutral or salt form. The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminium monostearate and gelatin. Sterile injectable solutions are prepared by incorporating the active polypeptides in the required amount in the appropriate solvent with several of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuumdrying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof. Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and in such amount as is therapeutically effective. The formulations are easily administered in a variety of dosage forms, such as the type of injectable solutions described above, but drug release capsules and the like can also be employed. For parenteral administration in an aqueous solution, for example, the solution should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, sterile aqueous media which can be employed will be known to those of skill in the art in light of the present disclosure. For example, one dosage could be dissolved in 1 ml of isotonic NaCl solution and either added to 1000 ml of hypodermoclysis fluid or inj ected at the proposed site of infusion. Some variation in dosage will necessarily occur depending on the condition of the subject being treated. The person responsible for administration will, in any event, determine the appropriate dose for the individual subject.
Thus, in another aspect, the invention refers to a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolving El compound (RvEl), resolving E2 compound (RvE2), resolving E3 compound (RvE3), resolving E4 compound (RvE4), resolving DI compound (RvDl), resolving D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
Thus, in another aspect, the invention refers to a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolving El compound (RvEl), resolving E2 compound (RvE2), resolving E3 compound (RvE3), resolving E4 compound (RvE4), resolving D2 compound (RvD2), resolving D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5).
In some embodiments, the pharmaceutical composition comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 SPM compounds selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In some embodiments, the pharmaceutical composition comprises 1, 2, 3, 4, 5, 6, 7, 8 or 9 SPM compound is selected in the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5) and combinations thereof.
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl) and lipoxin A4 compound (LXA4).
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl) and/or resolvin D3 compound (RvD3).
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl) and lipoxin B4 compound (LXB4).
In some embodiments, the pharmaceutical composition comprises resolvin E2 compound (RvE2) and/or protectin compound (PD1).
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl) and/or protectin compound (PD1). In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl) and/or resolvin D3 compound (RvD3).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl) and/or resolvin D5 compound (RvD5).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
In some embodiments, the pharmaceutical composition comprises Lipoxin A4 compound (LXA4) and protectin compound (PD1).
In some embodiments, the pharmaceutical composition comprises lipoxin B4 compound (LXB4) and protectin compound (PD1).
In some embodiments, the pharmaceutical composition comprises lipoxin A4 compound (LXB4) and lipoxin A4 compound (LXA4).
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl), lipoxin B4 compound (LXB4) and/or protectin DI compound (PD1).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl), lipoxin A4 compound (LXA4) and protectin DI compound (PD1).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin El compound (RvEl), lipoxin B4 compound (LXB4) and lipoxin A4 compound (LXA4).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin DI compound (RvDl), resolvin D3 compound (RvD3) and resolvin El compound (RvEl).
In some embodiments the pharmaceutical composition comprises at least one SPMs are resolvin DI compound (RvDl), resolvin D5 compound (RvD5) and resolvin El compound (RvEl).
In some embodiments the pharmaceutical composition comprises at least one SPMs are protectin DI compound (PD1), and/or resolvin D5 compound (RvD5) and/or resolvin El compound (RvEl).
In some embodiments the pharmaceutical composition comprises at least one SPMs are protectin DI compound (PD1), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4). In some embodiments, the pharmaceutical composition comprises resolvin El (RvEl) compound, lipoxin B4 compound (LXB4), resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
In some embodiments, the pharmaceutical composition comprises resolvin El compound (RvEl), lipoxin B4 compound (LXB4), lipoxin A4 compound (LXA4), resolvin D5 compound (RvD5) and/or protectin DI compound (PD1).
Thus, in another aspect, the invention refers to the pharmaceutical composition of the invention for use in therapy.
In particular embodiment, the invention refers to the pharmaceutical composition of the invention for use for treating or preventing Aspergillus fumigatus infection in patient in need thereof.
In particular embodiment, the invention refers to the pharmaceutical composition of the invention for use for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis.
Thus, in another aspect, the invention refers to a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis, wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, the invention refers to the pharmaceutical composition of the invention for use for treating cystic fibrosis in a patient in need thereof.
The invention refers to a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for treating cystic fibrosis in a patient in need thereof, wherein the at least one SPM compound is selected from the group consisting of protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
In particular embodiment, the invention refers to the pharmaceutical composition of the invention for use for repairing airway epithelial cell in patient suffering from cystic fibrosis.
The invention also refers to a pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for repairing airway epithelial cell in patient suffering from cystic fibrosis, wherein the at least one SPM compound is selected from the group consisting of r protectin DI compound (PD1), resolvin El compound (RvEl), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin DI compound (RvDl), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
The invention will be further illustrated by the following figures and examples. However, these examples and figures should not be interpreted in any way as limiting the scope of the present invention.
FIGURES:
Figure 1: Airway epithelial cells confluency over time. CFBE41O- cells were plated at 10 000 cells/well on a 96 wells plate and grown in EMEM culture medium without SVF for their synchronization at 37°C and 5% CO2 for 24h. Cells were then treated with SPMs (LXA4, LXB4, RvD2, PD1, RvEl and RvE2) or control solvent (CT EtOH). Images were captured every 2 hours for 84 hours and analyzed using the Incucyte technology.
Figure 2: Cell differentiation over time. TEER measurement of CFBE41O- cells (F508del CFTR) grown on permeable filter was followed during liquid/liquid (day 0 to day 20) and air/liquid) interface culture (from day 20). Cells were treated or not with LXA4, LXB4 or RvEl (lOnM) in the basolateral culture compartment the day of plating and the SPM treatment was renewed every second day.
Figure 3: Confocal images of ZO-1 (white) in CFBE41o- cell line. Asperigillus fumigatus (Af) exposure significantly disrupted tight junction organization compared to control condition. LXA4, LXB4, RvEl, PD1 and RvD5 treatment simultaneously with Af exposure prevented ZO-1 disruption. Figure 4: Quantification of ZO-1 in CFBE41O- cell line. Asperigillus fumigatus (Af) exposure significantly disrupted tight junction organization compared to control condition. LXA4, LXB4, RvEl, PD1 and RvD5 treatment at 10 nM simultaneously with Af exposure prevented ZO-1 disruption (n=5).
Figure 5: Implication of different receptors in the effect of RvEl, PD1 and Rvd5 on the tight junction organization. Chemerin inhibited the effect of RvEl in preventing tight junction disruption during 4/' exposure, suggesting the competitive effect of chemerin and RvEl on chemR23 receptor. Txl4A inhibited the effect of PD1, suggesting the role of GPR37 receptor in PD1 effect on tight junction protection. The AntiGPR32 inhibited the effect of RvD5, suggesting a role for GPR32 receptor in the effect of RvD5 on tight junction.
Figure 6: SPMs reduce the Asperigillus fumigatus growth in human nasal epithelial cell (hNEC). The Af growth was significantly reduced when Af conidies were inoculated on hNEC (compared with its growth without epithelial cells). PD1 and LXA4 significantly enhanced this effect
EXAMPLE:
Material & Methods
The CFBE41o- bronchial epithelial cell line over-expressing F508del CFTR mutation or human nasal epithelial cell primary cultures from patients with CF were cultured under airliquid interface to obtain a differentiated epithelium. Af expressing were inoculated at a MOI of 1:6 on the apical side of the epithelium with or without SPMs (lOnM) treatment. The tight junction protein (ZO-1) was stained by immunofluorescence. The Airway surface liquid layer (ASL) was stained using Texas red. Confocal microscopy was used to visualize 4/locati on and tight junction integrity and ASL.
Results:
Impact of SPMs on CF bronchial epithelial cell proliferation
Exposure to LXA4, LXB4, RvD2, RvD3, RvD4, PD1, RvEl and RvE2 (lOnM) significantly stimulated airway epithelial cells to reach confluency (n=25). In control conditions (EtOH) 80% of confluency was reached after 64 hours growth while cells treated with SPMs reached the same confluency much earlier: LXB4 after 50 hours; RvD2, PD1, RvEl and RvE2 after 52 hours; LXA4 after 54 hour (Figure 1). During treatment with RvD3 and RvD4 confluency was reached after 56 hours and RvE4 after 60 hours (not shown). Treatment with RvDl, RvD5 did not produce any effect on cell growth compared to control (not shown). Impact of SPMs in CF bronchial epithelial cell barrier function during differentiation
Cells barrier integrity was assessed during cell differentiation and tight junction formation using Trans Epithelial Electrical Resistance (TEER) measurement. In control condition (CT EtOH) TEER increased in two phase, one fast increase from the day of plating cells to Day 10, followed by a further and slower increase until Day 20. At Day 20, the apical medium was removed to continue cell culture at air/liquid interface. At this later stage, the differentiation is TEER decreased as a result of ion transport establishment. The 3 SPMs used, LXA4, LXB4 and RvEl (lOnM) significantly enhanced a TEER increase over time during the first phase of differentiation, which correspond to the tight junction formation (Figure 2).
Impact of SPMs in protecting CF bronchial epithelial cells from Aspergillus fumigatus Aspergillus fumigatus (Al), a predominant fungus in CF patient’s airways is associated with lung injury and lung function decline. Here, we assessed the impact of SPMs during Af exposure of CF bronchial epithelial cells.
Af transmigrated from the apical to the basolateral side of the airway epithelial layer within 24h. Airway epithelial cells exposed to Af showed strong cellular shape damages and significantly altered tight junctions. LXA4, LXB4, RvEl, PD1 or RvD5 treatment (lOnM) and prevented ZO-1 disruption induced by cell exposure to Af (figure 3 and figure 5).
Impact of SPMs on the airway surface liquid layer height
CF Airway epithelial showed a reduced airway surface liquid (ASL) layer height due to ion transport abnormalities. RvDl, RvD3 or RvEl used at lOnM restored the ASL height in CF airway epithelial cells (data not shown).
Implication of different receptors in the effect of RvEl, PD1 and Rvd5 on the tight junction organization.
The results are depicted in Figure 4. Chemerin inhibited the effect of RvEl in preventing tight junction disruption during 4/' exposure, suggesting the competitive effect of chemerin and RvEl on chemR23 receptor. Txl4A inhibited the effect of PD1, suggesting the role of GPR37 receptor in PD1 effect on tight junction protection. The AntiGPR32 inhibited the effect of RvD5, suggesting a role for GPR32 receptor in the effect of RvD5 on tight junction. It seems that different receptor are involved for RvEl, PD1 and RvD5, suggesting that these three SPMs will have additive effect when used in combination, especially to treat or prevent Af infection.
Impact of SPMs to reduce the Asperigillus fumigatus proliferation in human nasal epithelial cell (hNEC).
The results are depicted in Figure 6. The Af growth was significantly reduced when Af conidies were inoculated on hNEC (compared with its growth without epithelial cells). PD1 and LXA4 significantly enhanced this effect.
Conclusion:
Taken together, these data suggest a role for SPMs in maintaining mechanical and chemical barrier function and preventing tight junction disruption during Af infection of CF airway epithelial cells. These data are consistent with a role for abnormal SPM biosynthesis in the reduced ability of CF airway to fight infection and open new therapeutic perspectives. These data also suggest that SPMs stimulate the epithelial cell immune system and thus reduce the growth of Aspergillus fumigatus in CF patient.
REFERENCES:
Throughout this application, various references describe the state of the art to which this invention pertains. The disclosures of these references are hereby incorporated by reference into the present disclosure.
1. Sonneville F, Ruffin M, Coraux C, Rousselet N, Le Rouzic P, Blouquit-Laye S, et al.
TARGETING MIR-9 AND AN01: NEW THERAPEUTIC STRATEGY IN CYSTIC FIBROSIS. Pediatric Pulmonology 2017; 52: S291-S291.
2. Tabary O, Escotte S, Couetil JP, Hubert D, Dusser D, Puchelle E, et al. Relationship between
IkappaBalpha deficiency, NFkappaB activity and interleukin-8 production in CF human airway epithelial cells. Pflugers Arch 2001; 443 Suppl 1: S40-44.
3. Sonneville F, Ruffin M, Coraux C, Rousselet N, Rouzic PL, Blouquit-Laye S, et al.
MicroRNA-9 downregulates the ANO1 chloride channel and contributes to cystic fibrosis lung pathology. Nature Communications 2017; 8: 710.
4. GaliettaLJ, Haggie PM, Verkman AS. Green fluorescent protein-based halide indicators with improved chloride and iodide affinities. FEBS Lett 2001; 499: 220-224. . Serhan, "Treating inflammation and infection in the 21st century: new hints from decoding resolution mediators and mechanisms," (in eng), FASEB J, Jan 2017. CL, Flick LM, Park KW, Softie S, Greer TM, Keledjian R, Yang R, Uddin J, Guggino
WB, Atabani SF, Belkaid Y, Xu Y, Whitsett JA, Accurso FJ, Wills-Karp M, Petasis NA. Defective lipoxin-mediated anti-inflammatory activity in the cystic fibrosis airway.
Nat Immunol. 2004 Apr;5(4):388-92. . Ringholz et al., "Reduced 15-lipoxygenase 2 and lipoxin A4/leukotriene B4 ratio in children with cystic fibrosis," (in ENG), Eur Respir J, Apr 2014. rbach, V. et al., “CF patients’ airway epithelium and sex contribute to biosynthesis defects of pro-resolving lipids” , Frontiers in Immunology, 2022 ickmeier et al., "Pro-resolving lipid mediator Resolvin DI serves as a marker of lung disease in cystic fibrosis," (in eng), PLoS One, vol. 12, no. 2, p. e0171249, 2017.

Claims

CLAIMS: 1. A method for treating cystic fibrosis in patient in need thereof comprising administering to said patient a therapeutically effective amount of at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin D1 compound (PD1), resolvin E1 compound (RvE1), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), and resolvin D5 compound (RvD5), 2. The method according to claim 1, wherein the at least one SPM repairs airway epithelial cell. 3. The method according to any claim 1, wherein the patient has or is susceptible to have Aspergillus fumigatus infection. 4. A method for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis comprising administering at least one specialized lipid mediator of the resolution of inflammation (SPM) compound selected from the group consisting of protectin D1 compound (PD1), resolvin E1 compound (RvE1), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D1 compound (RvD1), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4). 5. The method according to claim 1 to 3, wherein 1,
2,
3,
4,
5, 6, 7,8, 9, 10, 11 or 12 compound is selected in the group consisting of protectin D1 compound (PD1), resolvin E1 compound (RvE1), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D1 compound (RvD1), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
6. The method according to claim 4, wherein the compounds are resolvin El (RvEl) compound, lipoxin B4 compound (LXB4), protectin DI compound (PD1) and/or lipoxin A4 compound (LXA4).
7. The method according to claim 1 to 3, wherein the compounds are resolvin El (RvEl) compound, protectin DI (PD1) compound and/or resolving D5 (RvD5).
8. The method according to claim 1 or 3, wherein the patient subject harbors at least one allelic mutation selected from class I, class II, class III, class IV, class V or class VI.
9. The method of claim 6, wherein the subject harbors at least one mutation in the CFTR gene.
10. The method of claim 7, wherein the subject harbors at least one allelic mutation selected from F508del-CFTR, R117H CFTR, 2184delA CFTR, W1282X CFTR, 2183AA>G CFTR or G551D CFTR.
11. The method according to any claim 1 to 8, wherein the SPM compound is administered to the patient using any suitable method that enables the SPM compound to reach the lungs.
12. The method according to claim 9, wherein the at least one SPM compound is administered by inhalation.
13. The method according to any claim 1 to 10, wherein the at least one SPM compound is administered in combination with a classical treatment of cystic fibrosis.
14. The method according to claim 11, wherein the classical treatment is selected from the group consisting of tezcaftor, ivacaftor, lumacaftor, orkambi, lenabasum, elexacaftor, acebilusta and trikafta.
15. A pharmaceutical composition comprising at least one specialized lipid mediator of the resolution of inflammation (SPM) for use for treating or preventing Aspergillus fumigatus infection in patient suffering from cystic fibrosis, wherein the at least one SPM compound is selected from the group consisting of protectin D1 compound (PD1), resolvin E1 compound (RvE1), resolvin E2 compound (RvE2), resolvin E3 compound (RvE3), resolvin E4 compound (RvE4), resolvin D1 compound (RvD1), resolvin D2 compound (RvD2), resolvin D3 compound (RvD3), resolvin D4 compound (RvD4), resolvin D5 compound (RvD5), lipoxin A4 compound (LXA4) and lipoxin B4 compound (LXB4).
16. The pharmaceutical composition for use according to claim 15, comprising resolvin E1 compound (RvE1), protectin D1 compound (PD1) and/or resolving D5 compound (RvD5).
PCT/EP2023/065286 2022-06-07 2023-06-07 Specialised pro-resolution lipid mediator (spm) compounds for use in the treatment of cystic fibrosis and aspergillus fumigatus infection in patient suffering from cystic fibrosis WO2023237626A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP22305824 2022-06-07
EP22305824.9 2022-06-07

Publications (1)

Publication Number Publication Date
WO2023237626A1 true WO2023237626A1 (en) 2023-12-14

Family

ID=82308141

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2023/065286 WO2023237626A1 (en) 2022-06-07 2023-06-07 Specialised pro-resolution lipid mediator (spm) compounds for use in the treatment of cystic fibrosis and aspergillus fumigatus infection in patient suffering from cystic fibrosis

Country Status (1)

Country Link
WO (1) WO2023237626A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004110426A1 (en) * 2003-06-01 2004-12-23 Petasis Nicos A Modulation of airway inflammation in patients with cystic fibrosis and related diseases
WO2020109480A1 (en) * 2018-11-30 2020-06-04 Evonik Operations Gmbh Preparation for use to increase the formation of one or more specialized pro-resolving lipid mediators (spm)
WO2020115231A1 (en) * 2018-12-06 2020-06-11 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and compositions for treating cystic fibrosis airways
IT202000008251A1 (en) * 2020-04-17 2021-10-17 Univ Degli Studi G Dannunzio Di Chieti Pescara LIPID MOLECULES FOR USE IN THE TREATMENT OF PATHOLOGIES OF THE RESPIRATORY TRACT

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004110426A1 (en) * 2003-06-01 2004-12-23 Petasis Nicos A Modulation of airway inflammation in patients with cystic fibrosis and related diseases
WO2020109480A1 (en) * 2018-11-30 2020-06-04 Evonik Operations Gmbh Preparation for use to increase the formation of one or more specialized pro-resolving lipid mediators (spm)
WO2020115231A1 (en) * 2018-12-06 2020-06-11 INSERM (Institut National de la Santé et de la Recherche Médicale) Methods and compositions for treating cystic fibrosis airways
IT202000008251A1 (en) * 2020-04-17 2021-10-17 Univ Degli Studi G Dannunzio Di Chieti Pescara LIPID MOLECULES FOR USE IN THE TREATMENT OF PATHOLOGIES OF THE RESPIRATORY TRACT

Non-Patent Citations (19)

* Cited by examiner, † Cited by third party
Title
"GenBank", Database accession no. AC000061
"NCBI", Database accession no. P13569.3
AUDREZET ET AL., HUM. MUTAT., vol. 23, no. 4, 2004, pages 343 - 357
BRIOTTET M ET AL: "P133 Impact of specialised pro-resolving lipid mediators on cystic fibrosis airway epithelial cell functions", JOURNAL OF CYSTIC FIBROSIS, ELSEVIER, NL, vol. 20, 1 January 2021 (2021-01-01), XP086616044, ISSN: 1569-1993, [retrieved on 20210611], DOI: 10.1016/S1569-1993(21)01159-0 *
BRIOTTET MAELLE ET AL: "The Role of Specialized Pro-Resolving Mediators in Cystic Fibrosis Airways Disease", FRONTIERS IN PHARMACOLOGY, vol. 11, 2 September 2020 (2020-09-02), XP055982696, DOI: 10.3389/fphar.2020.01290 *
C. N. SERHAN: "Treating inflammation and infection in the 21st century: new hints from decoding resolution mediators and mechanisms", FASEB J, January 2017 (2017-01-01)
CAS , no. 1025684-60-9
ELBOM JS.: "Cystic fibrosis", SEMINAR, 2016
F. C. RINGHOLZ ET AL.: "Reduced 15-lipoxygenase 2 and lipoxin A4/leukotriene B4 ratio in children with cystic fibrosis", EUR RESPIR J, April 2014 (2014-04-01)
GALIETTA LJHAGGIE PMVERKMAN AS: "Green fluorescent protein-based halide indicators with improved chloride and iodide affinities", FEBS LETT, vol. 499, 2001, pages 220 - 224, XP004247019, DOI: 10.1016/S0014-5793(01)02561-3
HARRIET C.KRISTIN E. T.OLIVIER T. ET AL.: "Translating the genetics of cystic fibrosis to personalized medicine", TRANSL RES, vol. 168, 2016, pages 40 - 49, XP029385950, DOI: 10.1016/j.trsl.2015.04.008
JUN YANG ET AL: "Metabolomic profiling of regulatory lipid mediators in sputum from adult cystic fibrosis patients", FREE RADICAL BIOLOGY & MEDICINE, ELSEVIER INC, US, vol. 53, no. 1, 1 May 2012 (2012-05-01), pages 160 - 171, XP028491640, ISSN: 0891-5849, [retrieved on 20120508], DOI: 10.1016/J.FREERADBIOMED.2012.05.001 *
KARP CL, FLICK LM, PARK KW, SOFTIC S, GREER TM, KELEDJIAN R, YANG R, UDDIN J, GUGGINO WB, ATABANI SF, BELKAID Y, XU Y, WHITSETT JA: "Defective lipoxin-mediated anti-inflammatory activity in the cystic fibrosis airway.", NAT IMMUNOL., vol. 5, no. 4, April 2004 (2004-04-01), pages 388 - 92, XP002378278, DOI: 10.1038/ni1056
O. EICKMEIER ET AL.: "Pro-resolving lipid mediator Resolvin D1 serves as a marker of lung disease in cystic fibrosis", PLOS ONE, vol. 12, no. 2, 2017, pages e0171249, XP055611790, DOI: 10.1371/journal.pone.0171249
RECCHIUTI A ET AL: "382: SARS-CoV-2 induces distinct inflammatory signals in cystic fibrosis macrophages that are seduced by resolvins", JOURNAL OF CYSTIC FIBROSIS, ELSEVIER, NL, vol. 20, 15 October 2021 (2021-10-15), XP086829300, ISSN: 1569-1993, [retrieved on 20211015], DOI: 10.1016/S1569-1993(21)01806-3 *
RECCHIUTI ANTONIO ET AL: "Roles, Actions, and Therapeutic Potential of Specialized Pro-resolving Lipid Mediators for the Treatment of Inflammation in Cystic Fibrosis", FRONTIERS IN PHARMACOLOGY, vol. 10, 2 April 2019 (2019-04-02), XP055982736, DOI: 10.3389/fphar.2019.00252 *
SONNEVILLE F, RUFFIN M, CORAUX C, ROUSSELET N, ROUZIC PL, BLOUQUIT-LAYE S: "MicroRNA-9 downregulates the ANO1 chloride channel and contributes to cystic fibrosis lung pathology.", NATURE COMMUNICATIONS, vol. 8, 2017, pages 710, XP055428029, DOI: 10.1038/s41467-017-00813-z
TABARY O, ESCOTTE S, COUETIL JP, HUBERT D, DUSSER D, PUCHELLE E: "Relationship between IkappaBalpha deficiency, NFkappaB activity and interleukin-8 production in CF human airway epithelial cells.", PFLUGERS ARCH, vol. 443, no. 1, 2001, pages 40 - 44
V. URBACH, V. ET AL.: "CF patients' airway epithelium and sex contribute to biosynthesis defects of pro-resolving lipids", FRONTIERS IN IMMUNOLOGY, 2022

Similar Documents

Publication Publication Date Title
US20220233639A1 (en) Polypeptide for use in the protection of oxygen sensitive gram-positive bacteria
JP7082162B2 (en) Meibomian gland dysfunction treatment
Moss Cystic fibrosis: pathogenesis, pulmonary infection, and treatment
JP2007269778A (en) Method for treatment of macular degeneration and related eye conditions
WO2020115231A1 (en) Methods and compositions for treating cystic fibrosis airways
EP1392355B1 (en) Use of proteasome inhibitors to treat dry eye disorders
EP2732817B1 (en) A compound for use in treating injury associated with exposure to phosgene or chlorine gas
AU2002308760A1 (en) Use of proteasome inhibitors to treat dry eye disorders
WO2023237626A1 (en) Specialised pro-resolution lipid mediator (spm) compounds for use in the treatment of cystic fibrosis and aspergillus fumigatus infection in patient suffering from cystic fibrosis
US20210189390A1 (en) Combination treatment for cystic fibrosis
US20240065994A1 (en) Compositions and methods for preventing or treating diseases or disorders associated with protein misfolding
US20230375576A1 (en) Methods for the treatment of familial heterozygous and homozygous hypercholesterolemia with cyclodextrins
JP2007224030A (en) Method for treating macular degeneration and related eye symptom
WO2004110426A1 (en) Modulation of airway inflammation in patients with cystic fibrosis and related diseases
US11998562B2 (en) Ophthalmological formulations for the prevention of a coronavirus infection
US7655237B2 (en) Use of soluble CD14 for treatment of type 2 diabetes mellitus
US7112588B2 (en) Use of proteasome inhibitors to treat dry eye disorders
Kaya et al. Acetyl-leucine slows disease progression in lysosomal storage disorders Short title: Acetyl-leucine for lysosomal diseases
AU2021228371A1 (en) S1P receptor modulators
WO2022248542A1 (en) Methods and compositions for treating pulmonary alveolar proteinosis related to mars mutations
JP2006290772A (en) Therapeutic agent of multiple sclerosis
Manika et al. Νew treatments in Cystic Fibrosis
JP2000080030A (en) Agent for preventing and treating granuloma

Legal Events

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

Ref document number: 23731288

Country of ref document: EP

Kind code of ref document: A1