ITFE20080032A1 - ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM - Google Patents
ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM Download PDFInfo
- Publication number
- ITFE20080032A1 ITFE20080032A1 IT000032A ITFE20080032A ITFE20080032A1 IT FE20080032 A1 ITFE20080032 A1 IT FE20080032A1 IT 000032 A IT000032 A IT 000032A IT FE20080032 A ITFE20080032 A IT FE20080032A IT FE20080032 A1 ITFE20080032 A1 IT FE20080032A1
- Authority
- IT
- Italy
- Prior art keywords
- miglustat
- cystic fibrosis
- cells
- inflammatory
- cellule
- Prior art date
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
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
- A61K31/445—Non condensed piperidines, e.g. piperocaine
-
- 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
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Molecular Biology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Description
DESCRIZIONE dell'invenzione industriale dal titolo: DESCRIPTION of the industrial invention entitled:
"Effetto antiinfiammatorio del Miglustat su cellule dell'epitelio bronchiale" "Anti-inflammatory effect of Miglustat on bronchial epithelial cells"
DESCRIZIONE DESCRIPTION
La presente invenzione si riferisce all'impiego del Miglustat (N-butyldeoxynojirimycin), associata con il processo infiammatorio indotto in cellule epiteliali respiratorie da infezione con Pseudomonas aeruglnosa (P.aeruginosa); si propone tale molecola per la preparazione di un medicamento per il trattamento terapeutico della fibrosi cistica (CF). The present invention relates to the use of Miglustat (N-butyldeoxynojirimycin), associated with the inflammatory process induced in respiratory epithelial cells by infection with Pseudomonas aeruglnosa (P.aeruginosa); this molecule is proposed for the preparation of a medicament for the therapeutic treatment of cystic fibrosis (CF).
La fibrosi cistica è una malattia genetica autosomica recessiva causata da mutazioni di un gene che codifica per una proteina canale di membrana denominata CFTR (da Cystic Fibrosis Transmembrane conductance Regulator). L'incidenza della malattia nella popolazione caucasica è di 1/2000-3000 nuovi nati, mentre è molto inferiore nei nativi africani ed asiatici (1). Cystic fibrosis is an autosomal recessive genetic disease caused by mutations in a gene that codes for a membrane channel protein called CFTR (from Cystic Fibrosis Transmembrane conductance Regulator). The incidence of the disease in the Caucasian population is 1 / 2000-3000 new born, while it is much lower in African and Asian natives (1).
L'impiego di molecole allo scopo di inibire il processo infiammatorio cronico nelle vie respiratorie di soggetti affetti da fibrosi cistica è effettuato da tempo. Nella maggior parte dei casi, tali molecole sono state applicate alla patologia infiammatoria cronica delle vie respiratorie nella fibrosi cistica dopo un uso consolidato in altre patologie infiammatorie croniche, sistemiche o locali, non necessariamente delle vie respiratorie. Le prime molecole utilizzate sono stati ì farmaci anti-ìnfiammatori corticosteroidei, quali il prednìsolone per via sistemica (2) oppure il flutìcasone propionato (3) ed il beclometasone (4) per vìa inalatoria. Successivamente, molecole anti-infiammatorie quali ibuprofene sono state largamente applicate (5,6), come pure macrolidi quali azìtromicina (7,8). Inoltre, allo scopo di ottenere un efficace effetto antiinfiammatorio, altre molecole sono state studiate a livello sperimentale, quali l'antiossidante N-acetilcisteina (9) e l'antagonista dei recettori dei leucotrieni montelukast (10), per citare i più noti. In sìntesi, in diversi casi l'effetto anti-infiammatorio è stato correlato in studi clinici controllati con un significativo miglioramento oppure un rallentato decadimento della funzionalità respiratoria, mentre i principali limiti applicativi hanno riguardato l'insorgenza di effetti indesiderati sulla funzione corticosurrenale, oppure renale o gastrointestinale. The use of molecules in order to inhibit the chronic inflammatory process in the respiratory tract of subjects suffering from cystic fibrosis has been carried out for some time. In most cases, these molecules have been applied to chronic inflammatory pathology of the respiratory tract in cystic fibrosis after a consolidated use in other chronic, systemic or local inflammatory diseases, not necessarily of the respiratory tract. The first molecules used were corticosteroid anti-inflammatory drugs, such as systemic prednisolone (2) or fluticasone propionate (3) and beclomethasone (4) by inhalation. Subsequently, anti-inflammatory molecules such as ibuprofen have been widely applied (5,6), as well as macrolides such as azìtromycin (7,8). Furthermore, in order to obtain an effective anti-inflammatory effect, other molecules have been studied at an experimental level, such as the antioxidant N-acetylcysteine (9) and the antagonist of leukotriene receptors montelukast (10), to name the best known. In other words, in several cases the anti-inflammatory effect has been correlated in controlled clinical studies with a significant improvement or a slowed decay of respiratory function, while the main application limitations concerned the onset of undesirable effects on adrenal or renal function. or gastrointestinal.
Nell'individuo affetto da fibrosi cistica, il processo infiammatorio è associato all'infezione inizialmente ricorrente ad opera di differenti specie batteriche, tra le quali Staphylococcus aureus ed Haemophilus influenzae, seguita da colonizzazione cronica da P. aeruginosa. In taluni casi si instaurano anche infezioni croniche sostenute da Burkholderia cepacia, da micobatteri e da funghi quali 1'Aspergillus fumigatus. Il processo infiammatorio polmonare nella fibrosi cistica è caratteristicamente sostenuto da un infiltrato cellulare prevalentemente costituito da neutrofili polimorfonucleati, associato a significativo aumento dell'espressione del gene per la citochina interleuchina-8 (IL-8), presente in concentrazioni aumentate nel lavaggio broncoalveolare di pazienti fibrocistici ancor prima dell'instaurarsi di infezioni batteriche (11, 12, 13) ed ulteriormente aumentata dopo l'insorgenza di infezione polmonare batterica (14, 15). IL-8 è una delle più potenti chemochine, componente essenziale del sistema difensivo dell'ospite, rilasciata dalle cellule dell'epitelio respiratorio e da cellule dell'immunità in risposta ai batteri per attivare principalmente la migrazione extravasale e transepiteliale (16), che nella fibrosi cistica porta all'accumulo di un numero abnorme di leucociti neutrofili nella mucosa e nel lume di bronchi e bronchioli (1). In the individual with cystic fibrosis, the inflammatory process is associated with initially recurrent infection by different bacterial species, including Staphylococcus aureus and Haemophilus influenzae, followed by chronic colonization by P. aeruginosa. In some cases, chronic infections sustained by Burkholderia cepacia, by mycobacteria and by fungi such as Aspergillus fumigatus are also established. The pulmonary inflammatory process in cystic fibrosis is characteristically supported by a cell infiltrate predominantly consisting of polymorphonuclear neutrophils, associated with significantly increased expression of the cytokine interleukin-8 (IL-8) gene, present in increased concentrations in bronchoalveolar lavage of patients fibrocytes even before the onset of bacterial infections (11, 12, 13) and further increased after the onset of bacterial lung infection (14, 15). IL-8 is one of the most potent chemokines, an essential component of the host's defensive system, released by respiratory epithelial cells and immunity cells in response to bacteria to mainly activate extravasal and transepithelial migration (16), which in cystic fibrosis leads to the accumulation of an abnormal number of neutrophilic leukocytes in the mucosa and lumen of the bronchi and bronchioles (1).
In questa sede, i neutrofili liberano acidi nucleici, che contribuiscono alla riduzione della clearance mucociliare, e proteasi, responsabili del danno tessutale della parete bronchiale e della fibrosi peribronchiolare che possono inoltre portare alla riduzione della capacità battericidica dei neutrofili stessi (17). Ne consegue che il richiamo massivo di neutrofili nel lume bronchiale non soltanto risulta inefficace nell'eliminare i batteri ma è anche la causa principale del graduale peggioramento della funzione polmonare nei pazienti FC (18,19). Here, the neutrophils release nucleic acids, which contribute to the reduction of mucociliary clearance, and proteases, responsible for the tissue damage of the bronchial wall and peribronchiolar fibrosis which can also lead to the reduction of the bactericidal capacity of the neutrophils themselves (17). It follows that the massive recall of neutrophils into the bronchial lumen is not only ineffective in eliminating bacteria but is also the main cause of the gradual worsening of lung function in CF patients (18,19).
Una terapia diretta a ridurre l'espressione di IL-8 nel trattamento di pazienti affetti da fibrosi cistica potrebbe pertanto ridurre gli effetti negativi indotti dal processa infiammatorio. A therapy aimed at reducing the expression of IL-8 in the treatment of patients with cystic fibrosis could therefore reduce the negative effects induced by the inflammatory process.
Alla base della presente invenzione vi è pertanto la necessità di nuovi modificatori del processo di trascrizione del gene IL-8, utilizzabili nel trattamento della fibrosi cistica, che presentino un elevato livello di regolazione negativa dell'espressione dei geni IL-8 e un basso livello di citotossicità. At the basis of the present invention there is therefore the need for new modifiers of the transcription process of the IL-8 gene, usable in the treatment of cystic fibrosis, which present a high level of negative regulation of the expression of IL-8 genes and a low level of cytotoxicity.
Il Miglustat è un noto inibitore della biosintesi di glicolipidi (20). Oltre ad inibire la biosintesi di glicolipidi, il miglustat e' anche un potente inibitore dell'enzima a-glucosidasi (21). Recentemente è stato dimostrato che il miglustat corregge la funzione di trasporto degli ioni cloro della proteina F508del~CFTR in cellule epiteliali FC umane e murine (22). Noi abbiamo osservato che il miglustat è un potente inibitore del processo infiammatorio sia in vitro che in vivo. Miglustat is a known inhibitor of glycolipid biosynthesis (20). In addition to inhibiting glycolipid biosynthesis, miglustat is also a potent inhibitor of the a-glucosidase enzyme (21). Miglustat has recently been shown to correct the chlorine ion transport function of the F508del ~ CFTR protein in human and murine FC epithelial cells (22). We have observed that miglustat is a potent inhibitor of the inflammatory process both in vitro and in vivo.
La formula di struttura del miglustat è la seguente: The structural formula of miglustat is as follows:
In particolare, abbiamo trovato che il miglustat inibisce l'accumulo di mRNA per IL-8 in vitro cellule epiteliali bronchiali IB3-1 e CuFi in seguito ad infezione con P.aeruginosa o a stimoli con TNF-alfa, IL-1beta. IB3-1 è una linea cellulare derivata dall'epitelio bronchiale, immortalizzata con adenol2 /SV40 e derivata da un paziente CF con genotipo F508del/W1282X (23). Le cellule CuFi-1 derivano dall'epitelio bronchiale di un paziente con genotipo F508del omozigote (24). Gli studi in vivo sono stati effettuati in topi C57BL/6J ai quali sono state somministrate 3 dosi di miglustat 72, 48 and 24 ore prima dello stimolo intranase con LPS. Abbiamo osservato che il miglustat riduce sensibilmente il numero di granulociti neutrofili presenti nel liquido di lavaggio bronchìo-alveolare, recuperato 4 ore dopo lo stimolo con LPS In particular, we found that miglustat inhibits IL-8 mRNA accumulation in vitro IB3-1 and CuFi bronchial epithelial cells following infection with P.aeruginosa or stimulation with TNF-alpha, IL-1beta. IB3-1 is a cell line derived from bronchial epithelium, immortalized with adenol2 / SV40 and derived from a CF patient with genotype F508del / W1282X (23). CuFi-1 cells are derived from the bronchial epithelium of a patient with homozygous F508del genotype (24). In vivo studies were performed in C57BL / 6J mice administered 3 doses of miglustat 72, 48 and 24 hours prior to intranase stimulation with LPS. We observed that miglustat significantly reduces the number of neutrophilic granulocytes present in the bronchial-alveolar lavage fluid, recovered 4 hours after stimulation with LPS.
Dal punto di vista dell'applicazione clinica, il Miglustat è stato proposto per il trattamento della malattia di Gaucher di tipo I (25). Le informazioni sulla farmacocinetica e sugli effetti secondari del Miglustat sono già disponibili nella letteratura corrente (26). From the point of view of clinical application, Miglustat has been proposed for the treatment of type I Gaucher disease (25). Information on the pharmacokinetics and side effects of Miglustat is already available in the current literature (26).
La sintesi chimica dì Miglustat e di analoghi strutturali è stata descritta da diversi gruppi di ricerca (si vedano ad esempio i riferimenti bibliografici citati in 27). The chemical synthesis of Miglustat and structural analogues has been described by several research groups (see for example the bibliographical references cited in 27).
Un primo oggetto della presente invenzione è pertanto l'impiego di Miglustat e suoi analoghi strutturali per la preparazione di un medicamento per il trattamento terapeutico della fibrosi cistica. A first object of the present invention is therefore the use of Miglustat and its structural analogues for the preparation of a medicament for the therapeutic treatment of cystic fibrosis.
D'altra parte, un trattamento combinato con diversi modificatori del processo infiammatorio potrebbe permettere di incrementare la risposta biologica e clinica alla terapia. On the other hand, a combined treatment with different modifiers of the inflammatory process could allow to increase the biological and clinical response to therapy.
Perciò, un secondo oggetto della presente invenzione è l'impiego di Miglustat e analoghi strutturali in combinazione con almeno un ulteriore molecola ad azione anti-infiammatoria per la preparazione di un medicamento per il trattamento della fibrosi cistica. Therefore, a second object of the present invention is the use of Miglustat and structural analogues in combination with at least one further anti-inflammatory molecule for the preparation of a medicament for the treatment of cystic fibrosis.
Secondo una forma di attuazione preferita, detto ulteriore modificatore del processo infiammatorio è scelto dal gruppo che consiste di azitromicina e di ibuprofene. According to a preferred embodiment, said further modifier of the inflammatory process is selected from the group consisting of azithromycin and ibuprofen.
L'attività di Miglustat come inibitore del processo infiammatorio è stata valutata sia in cellule fibrocìstiche in coltura che in vivo in modelli murini di infiammazione polmonare. I risultati di tale studio sono illustrati negli esempi riportati nelle Tabelle che seguono. The activity of Miglustat as an inhibitor of the inflammatory process was evaluated both in cultured fibrocystic cells and in vivo in murine models of lung inflammation. The results of this study are illustrated in the examples shown in the following Tables.
Gli esempi che seguono vengono forniti a scopo di illustrazione e non sono intesi a limitare in alcun modo a portata dell'invenzione. The following examples are provided for the purpose of illustration and are not intended to limit the scope of the invention in any way.
ESEMPIO 1 EXAMPLE 1
Effetto di Miglustat sull'espressione di mediatori del processo infiammatorio in cellule IB3-1 s CuFi-1 stimolate con P.aeruginosa. Effect of Miglustat on the expression of inflammatory process mediators in IB3-1 s CuFi-1 cells stimulated with P.aeruginosa.
Questo esperimento è stato effettuato per valutare l'effetto del miglustat sull'espressione di meditori del processo infiammatorio indotto in cellule epiteliali respiratorie da P. aeruginosa. Miglustat riduce significativamente l'accumulo sia di IL-8 (p<0.0001)che di ICAM-1(p<0.0009)in cellule IB3-1. Analogamente il miglustat ha lo stesso effetto in cellule CuFi-1 dove inibisce l'espressione sia di IL-8 (p<0.003) che di ICAM-1 (p<0.03). Questi risultati dimostrano un potenziale effetto anti-infiammatorio del miglustat, come indicato dalla forte inibizione della trascrizione di IL-8 ed ICAM-1, due geni critici nella chemiotassi leucocitaria dopo infezione con P.aeruginosa. This experiment was performed to evaluate the effect of miglustat on the expression of mediators of the inflammatory process induced in respiratory epithelial cells by P. aeruginosa. Miglustat significantly reduces the accumulation of both IL-8 (p <0.0001) and ICAM-1 (p <0.0009) in IB3-1 cells. Similarly, miglustat has the same effect in CuFi-1 cells where it inhibits the expression of both IL-8 (p <0.003) and ICAM-1 (p <0.03). These results demonstrate a potential anti-inflammatory effect of miglustat, as indicated by the strong transcription inhibition of IL-8 and ICAM-1, two critical genes in leukocyte chemotaxis following infection with P.aeruginosa.
Figura 1. Miglustat riduce l'accumulo di mRNA di IL-8 e di ICAM -1, stimolato da P.aeruginosa in cellule IB3-1 e CuFi-1. Figure 1. Miglustat reduces the accumulation of IL-8 and ICAM -1 mRNA stimulated by P.aeruginosa in IB3-1 and CuFi-1 cells.
Le cellule sono state incubate con 200 microMolare miglustat per 24 ore e poi infettate con il ceppo di laboratorio PAO1. Colonie di PAO1 prelevate da una coltura overnight in TSA sono state inoculate in 20 ml di TSB e fatte crescere a 37°C, in agitazione fino a raggiungere l'assorbanza pari ad 1 OD alla lunghezza d'onda di 660 nm che è corrispondente a 1x10<9>CPU (colony forming units) /ml, come determinato in parallelo con il metodo della diluizione scalare. I batteri, lavati due volte con PBS, (phosphate buffered saline) sono stati risospesi e diluiti con LHC-8 e aggiunti alle cellule alla dose di 50 CFU/cell. L'infezione è stata effettuata incubando le cellule a 37°C per 4 ore. L'espressione relativa di mRNA (relativa rispetto a quella delle cellule noninfettate)e' ottenuta paragonando il rapporto IL-8/GAPDH (A, C) and ICAM-1/GAPDH (B,D) tra cellule infettate e non-infettate. I risultati sono espressi come media ± SEM di duplicati. L'analisi quantitativa Real-time RT-PCR è stata effettuata usando primers diretto e reverso per sequenze IL-8, ICAM-1 e il gene normalizzatore gliceraldeide-fosfato-deidrogenasi (GAPDH). I primers utilizzati sono stati: 5’-GTG GAG TCC ACT GGC GTC TT-3’ (GAPDH diretto), 5’-GCA AAT GAG CCC AGC CTT C-3<1>(GAPDH reverso), 5’-TAT GGC AAC GAC TCC TTC TCG-3' (ICAM-1 diretto), 5’-CTC TGC GGT CAC ACT GAC TGA-3’ (ICAM-1 reverso), 5'-GAC CAC ACT GCG CCA ACA-3' (IL-B diretto), 5'-GCT CTC TTC CAT CAG AAA GTT ACA TAA TTT-3' (IL-8 reverso. Cells were incubated with 200 microMolar miglustat for 24 hours and then infected with laboratory strain PAO1. PAO1 colonies taken from an overnight culture in TSA were inoculated in 20 ml of TSB and grown at 37 ° C, under agitation until the absorbance equal to 1 OD was reached at the wavelength of 660 nm which is corresponding to 1x10 <9> CPU (colony forming units) / ml, as determined in parallel with the scalar dilution method. The bacteria, washed twice with PBS, (phosphate buffered saline) were resuspended and diluted with LHC-8 and added to the cells at a dose of 50 CFU / cell. Infection was carried out by incubating the cells at 37 ° C for 4 hours. Relative mRNA expression (relative to that of uninfected cells) is obtained by comparing the ratio IL-8 / GAPDH (A, C) and ICAM-1 / GAPDH (B, D) between infected and non-infected cells. Results are expressed as mean ± SEM of duplicates. Quantitative Real-time RT-PCR analysis was performed using direct and reverse primers for IL-8 sequences, ICAM-1 and the glyceraldehyde-phosphate-dehydrogenase (GAPDH) normalizing gene. The primers used were: 5'-GTG GAG TCC ACT GGC GTC TT-3 '(GAPDH direct), 5'-GCA AAT GAG CCC AGC CTT C-3 <1> (GAPDH reverso), 5'-TAT GGC AAC GAC TCC TTC TCG-3 '(ICAM-1 direct), 5'-CTC TGC GGT CAC ACT GAC TGA-3' (ICAM-1 reverso), 5'-GAC CAC ACT GCG CCA ACA-3 '(IL-B direct), 5'-GCT CTC TTC CAT CAG AAA GTT ACA TAA TTT-3 '(IL-8 reverso.
ESEMPIO 2 EXAMPLE 2
Effetto di miglustat sull'espressione del mRNA di IL=8, stimolata da P.aeruginosa, TNFalpha o IL-lbeta in cellule IB3-1 e CuFi-1. Effect of miglustat on the mRNA expression of IL = 8, stimulated by P.aeruginosa, TNFalpha or IL-lbeta in IB3-1 and CuFi-1 cells.
In questo esperimento è stata valutata l'efficacia del miglustat nel ridurre l'accumulo di mRNA di IL-8 stimolato non soltanto da P.aeruginosa ma anche dalle citochine pro-infiammatorie TNF alpha e IL-lbeta, in cellule IB3-1 e CuFi-1. Il miglustat riduce sensibilmente l'espressione del mRNA di IL-8 non solo dopo stimolo con P.aeruginosa ma anche con TNF alpha o IL-1 beta, sia in cellule IB3-1 che CuFi-1. In this experiment, the efficacy of miglustat in reducing the accumulation of IL-8 mRNA stimulated not only by P.aeruginosa but also by pro-inflammatory cytokines TNF alpha and IL-lbeta, in IB3-1 and CuFi cells was evaluated. -1. Miglustat significantly reduces IL-8 mRNA expression not only after stimulation with P.aeruginosa but also with TNF alpha or IL-1 beta, both in IB3-1 and CuFi-1 cells.
Figura 2. Il miglustat riduce l'espressione del mRNA di IL-8 indotta da PAO1, o TNFalpha o IL-1beta in cellule IB3-1 e CuFi-1. Le cellule sono state incubate per 24 ore con miglustat (200 microMolare)e poi infettate con PAO1 come descritto in Figura 1 o stimolate con TNF alpha {10 ng/ml} o IL-1 beta (SO ng/ml) per 4 ore. I risultati sono espressi come % del valore di controllo ottenuto in cellule non trattate. Figure 2. Miglustat reduces PAO1, or TNFalpha, or IL-1beta-induced IL-8 mRNA expression in IB3-1 and CuFi-1 cells. Cells were incubated for 24 hours with miglustat (200 microMolar) and then infected with PAO1 as described in Figure 1 or stimulated with TNF alpha {10 ng / ml} or IL-1 beta (SO ng / ml) for 4 hours. The results are expressed as% of the control value obtained in untreated cells.
ESEMPIO 3 EXAMPLE 3
Effetto del miglustat in vivo Effect of miglustat in vivo
Qusto esperimento è stato condotto per vedere se l'azione anti-infiammatoria del miglustat, osservata in cellule fibrocistiche in coltura poteva essere estesa anche a modelli animali di infiammazione polmonare. -Abbiamo osservato che il miglustat riduce sensibilmente il numero di granulociti neutrofili presenti nel liquido di lavaggio bronchio-alveolare, recuperato 4 ore dopo lo stimolo con LPS. This experiment was conducted to see if the anti-inflammatory action of miglustat observed in cultured fibrocystic cells could also be extended to animal models of lung inflammation. -We observed that miglustat significantly reduces the number of neutrophilic granulocytes present in the bronchoalveolar lavage fluid, recovered 4 hours after stimulation with LPS.
Figura 3. Il miglustat ha un'azione anti-infianmatoria in vivo. Gli studi in vivo sono stati effettuati in topi C57BL/6J ai quali sono state somministrate 3 dosi successive (2mg/dose) di miglustat 72, 48 and 24 ore prima dello stimolo intranase con LPS. Quindi gli animali sono stati sacrificati ed è stato recuperato il liquido di lavaggio bronchio-alveolare per la conta delle cellule. Lo stimolo intranasale con LPS induce un accumulo di cellule, principalmente granulociti neutrofili. Abbiamo osservato che il miglustat riduce significativamente il numero di granulociti neutrofili presenti nel liquido di lavaggio bronchio-alveolare, recuperato 4 ore dopo lo stimolo con LPS. Figure 3. Miglustat has anti-inflammatory action in vivo. In vivo studies were performed in C57BL / 6J mice administered 3 successive doses (2mg / dose) of miglustat 72, 48 and 24 hours prior to intranase stimulation with LPS. Then the animals were sacrificed and the bronchoalveolar lavage fluid was recovered for cell counting. Intranasal stimulation with LPS induces an accumulation of cells, mainly neutrophilic granulocytes. We observed that miglustat significantly reduces the number of neutrophilic granulocytes present in the bronchoalveolar lavage fluid, recovered 4 hours after LPS stimulation.
Bibliografia Bibliography
1. Welsh JM, Ramsey BW, Accurso F, Cutting GR. Cystic Fibrosis in «The Metabolic and Molecular Bases of Inherited Diseases». Scriver CR, Beaudet AL, Sly WS, Valle D (Eds) McGraw-Hill, New York, 2001 1. Welsh JM, Ramsey BW, Accurso F, Cutting GR. Cystic Fibrosis in "The Metabolic and Molecular Bases of Inherited Diseases". Scriver CR, Beaudet AL, Sly WS, Valle D (Eds) McGraw-Hill, New York, 2001
2. Eigen H, Rosenstein BJ, FitzSimmons S, Schidlow DV.A multicenter study of alternate-day prednisone therapy in patients with cystic fibrosis. Cystic Fìbrosis Foundation Prednisone Trial Group.J Pediatr. 126:515-23, 1995 2. Eigen H, Rosenstein BJ, FitzSimmons S, Schidlow DV A multicenter study of alternate-day prednisone therapy in patients with cystic fibrosis. Cystic Fìbrosis Foundation Prednisone Trial Group.J Pediatr. 126: 515-23, 1995
3. Balfour-Lynn IM, Klein NJ, Dìnwiddie R. Randomised controlled trial of inhaled corticosteroids (fluticasone propionate)in cystic fibrosis. Arch Dis Child. 77:124-30, 1997 3. Balfour-Lynn IM, Klein NJ, Dìnwiddie R. Randomized controlled trial of inhaled corticosteroids (fluticasone propionate) in cystic fibrosis. Arch Dis Child. 77: 124-30, 1997
4. Wojtczak HA, Kerby GS, Wagener JS, Copenhaver SC,Gotlin RW, Riches DW, Accurso FJ. Beclomethasone diproprionate reduced airway inflammation without adrenal suppression in young children with cystic fibrosis: a pilot study. 4. Wojtczak HA, Kerby GS, Wagener JS, Copenhaver SC, Gotlin RW, Riches DW, Accurso FJ. Beclomethasone diproprionate reduced airway inflammation without adrenal suppression in young children with cystic fibrosis: a pilot study.
Pediatr Pulmonol. 32:293-302, 2001 Pediatr Pulmonol. 32: 293-302, 2001
5. Konstan MW, Byard PJ, Hoppel CL, Davis PB. Effect of high-dose ibuprofen in patients with cystic fibrosis . 5. Konstan MW, Byard PJ, Hoppel CL, Davis PB. Effect of high-dose ibuprofen in patients with cystic fibrosis.
N Engl J Med. 332:848-54, 1995 N Engl J Med. 332: 848-54, 1995
6. Lands LC, Milner R, Cantin AM, Manson D, Corey M. 6. Lands LC, Milner R, Cantin AM, Manson D, Corey M.
High-dose ibuprofen in cystic fibrosis: Canadian safety and effectiveness trial. J Pediatr. 151:249-54,2007 High-dose ibuprofen in cystic fibrosis: Canadian safety and effectiveness trial. J Pediatr. 151: 249-54,2007
7. Saiman L, Marshall BC, Mayer-Hamblett N, Burns JL, Quittner AL, Cibene DA,Coquillette S, Fieberg AY, Accurso FJ, Campbell PW 3rd; Macrolide Study Group. Azithromycin in patients with cystic fibrosisi chronically infected with Pseudomonas aeruginosa: a randomized controlled trial. JAMA. 290:1749-56, 2003 7. Saiman L, Marshall BC, Mayer-Hamblett N, Burns JL, Quittner AL, Cibene DA, Coquillette S, Fieberg AY, Accurso FJ, Campbell PW 3rd; Macrolide Study Group. Azithromycin in patients with cystic fibrosisi chronically infected with Pseudomonas aeruginosa: a randomized controlled trial. JAMA. 290: 1749-56, 2003
8. Clement A, Tamalet A, Leroux E, Ravilly S, Fauroux B, Jais JP. Long term effects of azithromycin in patients with cystic fibrosis: A doublé blind,__ placebo controlled trial. Thorax. 61:895-902, 2(306<’>8. Clement A, Tamalet A, Leroux E, Ravilly S, Fauroux B, Jais JP. Long term effects of azithromycin in patients with cystic fibrosis: A double blind, __ placebo controlled trial. Thorax. 61: 895-902, 2 (306 <'>
9. Tirouvanziam R, Conrad CK, Bottiglieri T, Herzenberg LA, Moss RB, Herzenberg LA. High-dose orai N-acetylcysteìne, a glutathione prodrug, modulates inflammation in cystic fibrosis. Proc Nati Acad Sci U SA. 103:4628-33,2006 9. Tirouvanziam R, Conrad CK, Bottiglieri T, Herzenberg LA, Moss RB, Herzenberg LA. High-dose now N-acetylcysteìne, a glutathione prodrug, modulates inflammation in cystic fibrosis. Proc Nati Acad Sci U SA. 103: 4628-33.2006
10. Schmitt-Grohé S, Eickmeier 0, Naujoks C, Schubert R, Lentze MJ, Zielen S,Rietschel E. Effects of longterm treatment with montelukast in mild cvstic fibrosis (long term treatment with montelukast in cystic fibrosis).Respir Med. 101:684,2007 10. Schmitt-Grohé S, Eickmeier 0, Naujoks C, Schubert R, Lentze MJ, Zielen S, Rietschel E. Effects of longterm treatment with montelukast in mild cvstic fibrosis (long term treatment with montelukast in cystic fibrosis). : 684,2007
11. Khan TZ, Wagener JS, Bost T, Martinez J, Accnrso FJ, Riches DW. Early pulmonary inflammation in infants with cystic fibrosis. Am J Respir Crit Care Med. 151J1075-82,1995 11. Khan TZ, Wagener JS, Bost T, Martinez J, Accnrso FJ, Riches DW. Early pulmonary inflammation in infants with cystic fibrosis. Am J Respir Crit Care Med. 151J1075-82,1995
12. Balough K, McCubbin M, Weinberger M, Smits W, Ahrens R, Fick R. The relationship between infection and inflammation in thè early stages of lung disease from cystic fibrosis. Pediatr Pulmonol. 20:63-70,1995 12. Balough K, McCubbin M, Weinberger M, Smits W, Ahrens R, Fick R. The relationship between infection and inflammation in the early stages of lung disease from cystic fibrosis. Pediatr Pulmonol. 20: 63-70,1995
13.Armstrong DS, Grimwood K, Carlin JB, Carzino R, Gutièrrez JP, Hull J, Olinsky A, Phelan EM, Robertson CF, Phelan PD. Lower airway inflammation in infants and young children with cystic fibrosis. Am J Respir Crrt Care Med. 156:1197<aT>204,1997 13. Armstrong DS, Grimwood K, Carlin JB, Carzino R, Gutièrrez JP, Hull J, Olinsky A, Phelan EM, Robertson CF, Phelan PD. Lower airway inflammation in infants and young children with cystic fibrosis. Am J Respir Crrt Care Med. 156: 1197 <aT> 204,1997
14.Bonfield TL, Panuska JR, Konstan MW, Hilliard KA, Hilliard JB, Ghnaim H, Berger M. Inflammatory cytokines in cystic fibrosis lungs. Am J Respir Crit Care Med. 152: 2111-2118,1995 14. Bonfield TL, Panuska JR, Konstan MW, Hilliard KA, Hilliard JB, Ghnaim H, Berger M. Inflammatory cytokines in cystic fibrosis lungs. Am J Respir Crit Care Med. 152: 2111-2118,1995
15.Noah TL, Black HR, Cheng PW, Wood RE, Leigh MW. 15.Noah TL, Black HR, Cheng PW, Wood RE, Leigh MW.
Nasal and bronchoalveolar lavage fluid cytokines in early cystic fibrosis. J Infect Dis. 175:638-47,1997 Nasal and bronchoalveolar lavage fluid cytokines in early cystic fibrosis. J Infect Dis. 175: 638-47, 1997
16.Mantovani A. Chemokines. Introduction and overview. 16.Mantovani A. Chemokines. Introduction and overview.
Chem Immunol. 72:1-6,1999 Chem Immunol. 72: 1-6.1999
17.Hartl D, et al. Cleavage of CXCR1 on neutrophils disables bacterial killing in cystic fibrosis lung disease. Nat Med. 13:1423-30,2007 17. Hartl D, et al. Cleavage of CXCR1 on neutrophils disables bacterial killing in cystic fibrosis lung disease. Nat Med. 13: 1423-30,2007
18.Dbring G and Worlitzsch D. Inflammation in cystic fibrosis and ìts management. Raediatr Respir Rev. 18.Dbring G and Worlitzsch D. Inflammation in cystic fibrosis and ìts management. Raediatr Respir Rev.
1:101-6, 2000 1: 101-6, 2000
19.Koehler DR et al. Lung inflammation as a therapeutic target in cystic fibrosis. Am J Respir Celi Mol Biol 31:377-381,2004 19.Koehler DR et al. Lung inflammation as a therapeutic target in cystic fibrosis. Am J Respir Celi Mol Biol 31: 377-381,2004
20.Platt FM, Neises GR, Dwek RA and Butters TD. N-butyldeoxynojirimycin is a novel inhibitor of glycolipid biosynthesis. J Biol Chem; 269: 8362-8365 1994 20. Platt FM, Neises GR, Dwek RA and Butters TD. N-butyldeoxynojirimycin is a novel inhibitor of glycolipid biosynthesis. J Biol Chem; 269: 8362-8365 1994
21. Butters TD, Dwek RA and Platt EM. Imino sugar inhibitors for treating thè lysosomal Glycoshingolipidoses. Glycobiology; 15, 43R-52R, 2005 21. Butters TD, Dwek RA and Platt EM. Imino sugar inhibitors for treating the lysosomal Glycoshingolipidoses. Glycobiology; 15, 43R-52R, 2005
22. Norez C, Noel S, Wilke M, Bijvelds M, Jorna H, Melin P, DEJonge H and Becq F. Rescue of functional delF508-CFTR channels in cystic fibrosis epithelial cells by thè a-glucosidase inhibitor miglustat. FEBS Lett.; 580:2081-2086, 2006 22. Norez C, Noel S, Wilke M, Bijvelds M, Jorna H, Melin P, DEJonge H and Becq F. Rescue of functional delF508-CFTR channels in cystic fibrosis epithelial cells by the a-glucosidase inhibitor miglustat. FEBS Lett .; 580: 2081-2086, 2006
23. Zeitlin PL, Lu L, Rhim J, Cutting G, Stetten G, Kieffer KA, Craig R, Guggino WB. A cystic fibrosis bronchial epithslial celi line: immortalization by adeno-12-SV40 infection. Am J Respir Celi Mol Biol. 23. Zeitlin PL, Lu L, Rhim J, Cutting G, Stetten G, Kieffer KA, Craig R, Guggino WB. A cystic fibrosis bronchial epithslial celi line: immortalization by adeno-12-SV40 infection. Am J Respir Celi Mol Biol.
4:313-9,1991 4: 313-9.1991
24. Zabner J, Karp P, Seiler M, Phillips SL, Mitchell CJ, Saavedra M, Welsh M, Klingelhutz AJ. Development of cystic fibrosis and noncystic fibrosis airway celi lines. Am J Physiol Lung Celi Mol Physiol. 24. Zabner J, Karp P, Seiler M, Phillips SL, Mitchell CJ, Saavedra M, Welsh M, Klingelhutz AJ. Development of cystic fibrosis and noncystic fibrosis airway celi lines. Am J Physiol Lung Celi Mol Physiol.
284:L844-54, 2003 284: L844-54, 2003
25.Cox T, Lachmann R, Hollak C, Aerts J, van Weely S, Hrebicek M, Platt F, Butters T, Dwek R, Moyses C, Gow I, Elstein D and Zimran A.() Nove! orai treatment of Gaucher's disease with N-butyldeoxynoj irimycin (OGT 918) to decrease substrate biosynthesis. Lancet; 355, 1481-1485, 2000 26.Cox TM et al .Advisory Council to thè Eruropean Working Group on Gaucher Disease. J Inherit Metab Dis.; 26:513-526, 2003 25.Cox T, Lachmann R, Hollak C, Aerts J, van Weely S, Hrebicek M, Platt F, Butters T, Dwek R, Moyses C, Gow I, Elstein D and Zimran A. () Nove! orai treatment of Gaucher's disease with N-butyldeoxynoj irimycin (OGT 918) to decrease substrate biosynthesis. Lancet; 355, 1481-1485, 2000 26.Cox TM et al. Advisory Council to the Eruropean Working Group on Gaucher Disease. J Inherit Metab Dis .; 26: 513-526, 2003
27.Asano N. Glycosidase inhibitors: update and perspectives on practical use. Glycobiology;13: 93R-104R, 2003 27.Asano N. Glycosidase inhibitors: update and perspectives on practical use. Glycobiology; 13: 93R-104R, 2003
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT000032A ITFE20080032A1 (en) | 2008-10-23 | 2008-10-23 | ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT000032A ITFE20080032A1 (en) | 2008-10-23 | 2008-10-23 | ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ITFE20080032A1 true ITFE20080032A1 (en) | 2010-04-24 |
Family
ID=41016790
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IT000032A ITFE20080032A1 (en) | 2008-10-23 | 2008-10-23 | ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM |
Country Status (1)
| Country | Link |
|---|---|
| IT (1) | ITFE20080032A1 (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005046672A2 (en) * | 2003-11-07 | 2005-05-26 | Centre National De La Recherche Scientifique | Use of glucosidase inhibitors for therapy of mucovisidosis |
| WO2008006007A2 (en) * | 2006-07-03 | 2008-01-10 | Columbia University | Defective cystic fibrosis transduction regulator (cftr) causes increased sphingolipid synthesis |
-
2008
- 2008-10-23 IT IT000032A patent/ITFE20080032A1/en unknown
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005046672A2 (en) * | 2003-11-07 | 2005-05-26 | Centre National De La Recherche Scientifique | Use of glucosidase inhibitors for therapy of mucovisidosis |
| WO2008006007A2 (en) * | 2006-07-03 | 2008-01-10 | Columbia University | Defective cystic fibrosis transduction regulator (cftr) causes increased sphingolipid synthesis |
Non-Patent Citations (5)
| Title |
|---|
| ANONYMOUS: "View of NCT00537602 on 2007_09_28", CLINICAL TRIALS.GOV ARCHIVE, 28 September 2007 (2007-09-28), XP002544444, Retrieved from the Internet <URL:http://clinicaltrials.gov/archive/NCT00537602/2007_09_28> * |
| DECHECCHI MARIA CRISTINA ET AL: "Anti-inflammatory effect of miglustat in bronchial epithelial cells.", JOURNAL OF CYSTIC FIBROSIS : OFFICIAL JOURNAL OF THE EUROPEAN CYSTIC FIBROSIS SOCIETY NOV 2008, vol. 7, no. 6, 23 September 2008 (2008-09-23), pages 555 - 565, XP002544443, ISSN: 1569-1993 * |
| EQUI A ET AL: "Long term azithromycin in children with cystic fibrosis: a randomised, placebo-controlled crossover trial", LANCET THE, LANCET LIMITED. LONDON, GB, vol. 360, no. 9338, 28 September 2002 (2002-09-28), pages 978 - 984, XP004794189, ISSN: 0140-6736 * |
| KONSTAN M W ET AL: "EFFECT OF HIGH-DOSE IBUPROFEN IN PATIENTS WITH CYSTIC FIBROSIS", NEW ENGLAND JOURNAL OF MEDICINE, THE, MASSACHUSETTS MEDICAL SOCIETY, WALTHAM, MA, US, vol. 332, 30 March 1995 (1995-03-30), pages 848 - 854, XP000951543, ISSN: 0028-4793 * |
| NOREZ ET AL: "Rescue of functional delF508-CFTR channels in cystic fibrosis epithelial cells by the alpha-glucosidase inhibitor miglustat", FEBS LETTERS, ELSEVIER, AMSTERDAM, NL, vol. 580, no. 8, 3 April 2006 (2006-04-03), pages 2081 - 2086, XP005352909, ISSN: 0014-5793 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1307189B1 (en) | Use of hydroxyethylrutosides for treating symptoms of common cold, allergic rhinitis and infections relating to the respiratory tract | |
| Proesmans et al. | What’s new in cystic fibrosis? From treating symptoms to correction of the basic defect | |
| US6407133B2 (en) | Treatment and prevention of reactive oxygen metabolite-mediated cellular damage | |
| ES2569391T3 (en) | Tritocualin for use in the treatment of cystic fibrosis | |
| ITMI20111068A1 (en) | TRIMETHYLANGELICINE AS A CFTR CORRECTOR IN BRONCHIAL EPITHELIUM CELLS | |
| WO2020115231A1 (en) | Methods and compositions for treating cystic fibrosis airways | |
| Gaylor et al. | Therapy with macrolides in patients with cystic fibrosis | |
| H Sears et al. | Treatment options for cystic fibrosis: state of the art and future perspectives | |
| US20240238217A1 (en) | Use of clofoctol for the treatment of inflammation | |
| ITFE20080032A1 (en) | ANTI-INFLAMMATORY EFFECT OF MIGLUSTAT CELLULE OF BRONCHIAL EPITHELIUM | |
| KR101297763B1 (en) | Use of S-adenosylmethionine(SAM) and superoxide dismutase(SOD) for the preparation of medicaments for the treatment of Alzheimer's disease | |
| ITFE20080033A1 (en) | ANTI-INFLAMMATORY EFFECT OF TRIMETHYL-ANGELICINE ON CELLS OF BRONCHIAL EPITHELIUM | |
| EP3983070B1 (en) | Method for treating asthma | |
| Chmiel et al. | Inflammation in the cystic fibrosis lung | |
| Dechecchi et al. | Pharmacological modulators of sphingolipid metabolism for the treatment of cystic fibrosis lung inflammation | |
| Thompson | Pharmacology of therapeutic aerosols | |
| US20240342227A1 (en) | Combination of e. faecalis and an anti-inflammatory agent and uses thereof in the prevention and/or treatment of respiratory diseases | |
| US20240408153A1 (en) | Bacteria useful in the prevention and/or treatment of respiratory pathologies | |
| Ohlemiller | A Question of Balance: Free Radicals and Cochlear Homeostasis | |
| EP4247350A1 (en) | Composition for use in a method for the prevention and treatment of diseases of the respiratory system | |
| KR20210103848A (en) | Pharmaceutical composition for alleviating or treating sepsis comprising hydroxyurea | |
| AU2014269143A1 (en) | Methods and compositions for the disruption of biofilms and treatment of disorders characterized by the presence of biofilms | |
| CN114057708A (en) | Traditional Chinese medicine monomer orientin for treating diabetic nephropathy | |
| Takeyama et al. | C35 ASTHMA AND ALLERGY CELLULAR INVESTIGATIONS: Theracurmin®, A Highly Absorptive Nanoparticle Curcumin, Reduces Th2 Inflammatory Responses And Airway Mucus Production In Ovalbumin-Induced Murine Model Of Asthma | |
| Amin et al. | Anti-inflammatory Therapies for Chronic Lung Diseases in Children |