EP1392729A2 - Use of a peptide which activates guanylate-cyclase c for the treatment of respiratory airway problems via the airways, medicament, inhalation devices and method of diagnosis - Google Patents
Use of a peptide which activates guanylate-cyclase c for the treatment of respiratory airway problems via the airways, medicament, inhalation devices and method of diagnosisInfo
- Publication number
- EP1392729A2 EP1392729A2 EP02745124A EP02745124A EP1392729A2 EP 1392729 A2 EP1392729 A2 EP 1392729A2 EP 02745124 A EP02745124 A EP 02745124A EP 02745124 A EP02745124 A EP 02745124A EP 1392729 A2 EP1392729 A2 EP 1392729A2
- Authority
- EP
- European Patent Office
- Prior art keywords
- peptide
- medicament
- peptides
- seq
- disorders
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/04—Linear peptides containing only normal peptide links
- C07K7/08—Linear peptides containing only normal peptide links having 12 to 20 amino acids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M15/00—Inhalators
- A61M15/0065—Inhalators with dosage or measuring devices
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4702—Regulators; Modulating activity
- C07K14/4705—Regulators; Modulating activity stimulating, promoting or activating activity
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
- A61M15/00—Inhalators
- A61M15/009—Inhalators using medicine packages with incorporated spraying means, e.g. aerosol cans
Definitions
- the invention relates to the use of a peptide which activates guanylate cyclase C for the treatment of respiratory diseases and disorders associated with ventilation disorders and / or disorders of mucus secretion, an associated medicament, an inhalation device and a method for diagnosing the aforementioned disorders.
- Obstructive ventilation disorders are a serious clinical problem. They are associated with a narrowing of the airways and thus an increase in flow resistance, spasms of the bronchial muscles, edematous swellings of the bronchial wall and increased secretion (hypercrine) of mucus with a tough consistency.
- the diseases associated with ventilation disorders and / or disorders of mucus secretion include Bronchial asthma, chronic bronchitis and cystic fibrosis.
- the invention has for its object to provide a new effective agent for the treatment of respiratory diseases and generally diseases that are associated with ventilation disorders and / or disorders of mucus secretion, which agent should allow liquefaction and better removal, especially of bronchial mucus.
- the object is achieved by the use of a peptide which activates guanylate cyclase C for the manufacture of a medicament for the treatment of respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion, via the airways, the medicament is formulated in such a way that the peptide is supplied on the air side of the respiratory tract, namely towards the apical membrane of the mucosal epithelial cells.
- peptides can also be administered together or in succession. Equivalent to the use of these peptides themselves is the use of homologous, essentially functionally identical peptides, in particular those peptide variants with the addition of single and / or several amino acids by deletion, insertion or exchange of single and / or several amino acids, and / or chemical derivatization (in particular the terminal amino acids) linked sequence modification.
- Pharmacologically acceptable derivatives are preferably amidated, acetylated, phosphorylated and glycosylated forms of the peptides and other post-translational derivatizations, including salts of these peptides and peptide derivatives.
- the peptide is in particular at least one of the peptides called guanylin, uroguanylin and lymphoguanylin or a heat-resistant enterotoxin. These peptides are known as such.
- a peptide homologous to the peptides mentioned and having essentially the same function can also be used.
- the homologs here are understood to mean those peptides which largely correspond to the sequences described below and are still attributed to the guanylin peptides by the person skilled in the art on the basis of their function and sequence homology. It is known to those skilled in the art that e.g. Point mutations, deletions and insertions do not have to affect the function of a peptide. Peptides modified in this way would therefore be classified as homologues.
- Seq. ID 1 (Guanylin, 15 AS): PGTCEICAYAACTGC Pro-Gly-Thr-Cys-Glu-Ile-Cys-Ala-Thr-Ala-Ala-Cys-Thr-Gly-Cys
- a 115 amino acid precursor molecule containing the above sequence is also often referred to as "guanylin". Both peptides are suitable for the purposes of the invention; the peptide with Seq is preferred. ID 1, which is a relatively small peptide that can be delivered via inhalation.
- a 15-AS peptide with the sequence PGTCEICAYAACTGC was first isolated from rat intestinal extracts and referred to as "guanylin". After cloning and characterization of the cDNA for the human Guanylin it was obvious that the Guanylin as a precursor molecule with 115 AS (SEQ ID 4. MNAFLLFALC LLGAWAALAG GVTVQDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKP LCKEPNAQEI LQRLEEIAED PGTCEICAYAACTGC) is synthesized. It is now known that it is not the precursor molecule that circulates in the blood as a bioactive protein, but the guanylin with 94 AS (Proguanylin 22-
- Human uroguanylin is a peptide to which the following amino acid sequences have been assigned:
- the uroguanylin was first isolated from the urine as a 16-AS peptide (NDDC ELCVNVACTG CL). The cloning and characterization of the cDNA for human uroguanylin resulted in a uroguanylin precursor molecule with 112 AS (Seq.
- Lymphoguanylin is a guanylin peptide expressed in lymphoid tissues, which has been described by Forte et al. was found (Forte et al. Endocrinology 1999, 140, 1800-1806). It is a 15 amino acid peptide with the following amino acid sequence:
- Seq. ID 3 (lymphoguanylin, 15 aa): QEECELCINMACTGY Gln-Glu-Glu-Cys-Glu-Leu-Cys-Ile-Asn-Met-Ala-Cys-Thr-Gly-Tyr
- the precursor molecule for lymphoguanylin contains 109 amino acids (SEQ ID 6: MKVLALPMAV TAMLLIL AQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNEGRELCINM
- lymphoguanylin come from the opossum.
- the human sequence is not yet known.
- the 15-amino acid lymphoguanylin also activates human guanylate cyclase C.
- guanylate cyclase a G protein-coupled receptor that catalyzes the formation of cyclic guanosine monophosphate (cGMP) from guanosine triphosphate (GTP).
- cGMP cyclic guanosine monophosphate
- GTP guanosine triphosphate
- guanylate cyclase activating peptides were discovered in succession, which are considered to be endogenous ligands for guanylate cyclase C. The first of these peptides was named guanylin (Currie, H.G. et al. Proc. Natl. Acad. Sei. USA 1992, 89, 947-951).
- the guanylate cyclase C thus not only acts as a receptor for the heat-stable enterotoxins, but it also represents the genuine receptor of the endogenous guanylin peptides.
- a sequence of a heat-stable enterotoxin which is suitable in the context of the invention is:
- these guanylin peptides listed above and the heat-stable enterotoxins lead to an increase in cGMP in the enterocytes via the activation of the common receptor.
- the increased cGMP level activates the cGMP-dependent protein kinase II (cGKII) in the enterocytes.
- cGKII cGMP-dependent protein kinase II
- This activated protein kinase phosphorylates and thereby opens the CFTR chloride channel in the apical membrane of the enterocytes. This results in the secretion of chloride ions and water into the lumen of the intestine.
- the CFTR chloride channel is now considered the final effector of the signal transduction chain of the guanylin peptides.
- These peptides are therefore a direct regulator of the CFTR chloride channel.
- the peptides mentioned circulate in the blood as endogenous activators. They can also be obtained from blood or haemofiltrate.
- DE 195 28 544 describes a guanylin peptide which was obtained from human blood and is intended for diagnostic, medical and commercial use as a medicament. This peptide was named GCAP-II. Due to the known effect of the guanylin peptides on guanylate cyclase C (see above), GCAP-II was specially designed for the treatment of diseases which are associated with disorders in the electrolyte transport in the cells. The application should preferably be by injection.
- guanylin The endogenous activator guanylin is found in various places in the body. Guanylin has been proven e.g. B. in the human pancreas (Kulaksiz et al, Histochem Cell Biol. (2001) 115, 131-145), in the kidney (Forte et al, Annu Rev. Physiol 2000, 62, 673-695), in the intestinal tract (Quian et al, Endocrinology 2000, 141, 3210-24) and in the lungs (Cetin et al, Proc. Natl. Acad. Sci. USA, 92, 5925 - 5929, 1995).
- the common receptor for heat-stable enterotoxins and guanylin peptides is located in the mucosa of the airways and is expressed there to a high degree on the apical membrane (air side) of the respective epithelial cells. but not on the basolateral membrane (blood side).
- the receptor located in the lungs can therefore not be stimulated via the bloodstream, but only via the airways.
- FIG. 1 shows schematically the signal transduction of the guanylin peptides on epithelial cells.
- GC-C Guanlyate cyclase
- GC-C is an enzyme-receptor complex that is localized as a membrane protein exclusively in the apical cell domain, which is directed towards the airway clearing. It lacks the basolateral membrane of the cells (blood side), which is known to be in contact with the circulating blood.
- GC-C Guanylin peptides that bind to the receptor (GC-C) via the airway clearing set in motion a specific intracellular mechanism that contains various protein modules.
- the GC-C activated externally by the guanylin peptides forms cGMP from GTP in high amounts intracellularly.
- This second messenger (cGMP) activates a membrane-associated cGMP-dependent protein kinase type II (cKGII), which carries out the phosphorylation and thus activation of the CFTR protein on its regulatory (R) domain.
- CFTR is a membrane protein in the apical membrane of the epithelial cells and is an important chloride channel that, after activation, secretes chloride ions from the cell towards the airway clearing.
- the water follows the secreted chlorine ions and flows into the clearing of the respiratory tract.
- the water comes from the epithelial cells and from the spaces between the cells (paracellular). Some of the chloride ions secreted into the clearing are taken up again in the cells; for this, bicarbonate ions are secreted from the cells. This exchange of ions is accomplished by the Type II (AE2) anion exchanger.
- AE2 Type II
- the AE2 protein is also located in the apical membrane of the epithelial cells.
- the bicarbonate ions are produced intracellularly by the enzyme carbonic anhydrase type II (CAM) from water and carbon dioxide.
- CAM carbonic anhydrase type II
- GC-C guanylate cyclase C
- cGKM cGMP-dependent protein kinase type II
- CFTR cystic fibrosis transmembrane conductance regulator
- AE-2 anion exchanger type 2
- CAM carbonic anhydrase type II.
- a central finding of the concept according to the invention is that the activation of the receptor by application of the endogenous ligands has to be carried out specifically via the airways.
- the person skilled in the art must therefore adjust the supply of the peptide or the medicament which contains the peptide in such a way that the peptide is fed - if possible exclusively - on the air side to the apical membrane of the respiratory tract and does not reach the bloodstream to a greater extent. This enables targeted local therapeutic use in the respiratory tract, especially since the receptor in the respiratory tract is located exclusively on the air side.
- the supply of the peptides according to the invention is a directed and direct supply to the receptor located on the air side.
- the appropriate means are available to the person skilled in the art for this. It can influence the directed supply to the air side via the adjustment of the peptide concentration in the pharmaceutical formulation, the dosage and the adjustment of the particle / droplet size within the formulation or the inhalation agent in such a way that practically no peptide on the blood side of the respiratory tract (to the basolateral membrane) and thus in enters the bloodstream.
- the optimal conditions for each selected peptide can be determined in targeted preliminary tests.
- the invention enables therapy with doses that are much lower than those that would be required to increase the blood concentration, while minimizing or eliminating the systemic side effects of the respective peptides.
- the peptides according to the invention act as stimulants in the sense of secretolysis by dissolving the viscous mucus present in the airways, the ion composition and the pH of the liquid being adjusted directly on the epithelial cells (“microclimate”) in such a way that the viscous mucus increasingly "liquefied”.
- the removal of mucus and microparticles from the respiratory tract is made possible by epithelial cells that have cilia on their apical side (air side).
- the "cleaning" function is achieved by beating the cilia.
- guanylin peptides in addition to their function of increasing the electrolyte and water secretion, in particular also activate the cilia-bearing epithelial cells, these cells have an increased beat frequency of the cilia. In the sense of a concerted action, the secretions and tiny particles on the mucous membrane of the respiratory tract are removed much more efficiently, which underlines the physiological and therapeutic importance of the guanylin peptides.
- the aforementioned newly found properties of the peptides according to the invention act synergistically in the sense of the invention and lead to the very good effect of the peptides supplied by the airways for the treatment of the disorder and diseases mentioned at the beginning.
- the peptides according to the invention can additionally be used for the production of diagnostics for respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion.
- the peptides themselves are suitable as reference substances for diagnostics.
- a lack / deficiency or an excess of these peptides, for example in bronchial mucus, exudate or lavage, can indicate the presence of disorders requiring treatment.
- the peptides can be detected using the customary and known means, such as spectrocopically, chromatographically or chemically.
- the person skilled in the art can use conventional methods and means to produce antibodies against the peptides according to the invention, which can then be used within molecular biological or enzymatic assays.
- To achieve the object of the invention therefore also contributes to a method for diagnosing the diseases mentioned, in which at least one of the peptides that activate guanylate cyclase C is detected, preferably in bronchial mucus, exudate, lavage, nasal secretions or saliva.
- the detection can be carried out by detecting one of the sequences for Seq. ID 1 to ID 6
- Seq. ID 4 (Guanylin precursor molecule): MNAFLLFALC LLGAWAALAG GV QDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKPLCKEPNAQEI LQRLEEIAED PGTCEICAYA ACTGC
- Seq. ID 2 (Uroguanylin): NDDC ELCVNVACTGCL Seq. ID 5 (Uroguanylin precursor molecule): MGCRAASGLLPGVAWLLLL LQSTQSVYIQ YQGFRVQLES MKKLSDLEAQ WAPSPRLQAQ SLLPAVCHHPALPQDLQPVC ASQEASSIFK TLRTIAN DDC ELCVNVACTG
- Seq. ID 6 (lymphoguanylin precursor molecule): MKVLALPMAVTAMLLILAQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNIFRAL RYIN QEECELCINM Seq. ID 7 (heat-stable enterotoxin): NSSNYCCELCCNPACTGCY (19 AS) from enteropathogenic E. coli.
- a positive test result for the detection of a disorder is evaluated if a concentration of at least one of the peptides that activate guanylate cyclase C that deviates from comparison samples of healthy volunteers is found.
- peptides according to the invention further consists in formulating a medicament which is supplied via the airways and contains at least one peptide which activates guanylate cyclase C.
- At least one further active ingredient and optionally auxiliaries and additives can be present in the drug.
- Other active ingredients are, for example, muscle relaxants, local antibiotics, primarily for the treatment of simultaneously grafted bacterial infections, or also additional mucolytics, secretolytics, antitussives or bronchodilating substances. The specialist will make the selection on the basis of the respective needs in the treatment of the diseases mentioned at the beginning.
- the drug can be prepared in solid or liquid form and is conveniently supplied by the user via the airways. For this purpose it can be administered with a commercially available nebulizer or inhalation device.
- the medicament is present as an inhalation agent and contains at least one propellant.
- Chlorofluorocarbons are particularly suitable as blowing agents. Suitable blowing agents are known to those skilled in the art. In general, all suitable aerosol formers or smoke formers can be used. Depending on the excipient, an aerosol or smoke is inhaled, with an aerosol being preferred.
- an inhalation device which contains the medicament, ie that the medicament is present in the inhalation device ready-made.
- Such an inhalation device can consist of a spray device, in particular a metering spray device or a metering inhaler (English: MDI, metered dose inhaler).
- MDI metered dose inhaler
- Suitable inhalers are known to the person skilled in the art and are described, for example, in US Pat. No. 3,915,165, EP 166476 and US Pat. No. 6,099,517.
- Ultrasonic nebulizers are also suitable.
- the peptides according to the invention should first be converted into a finely dispersed form for administration.
- compositions for this purpose, they can first be brought into solution or suspension and, if necessary, stabilized in this form with pharmaceutically acceptable additives.
- Compatible surfactants such as Tween ® can be used for stabilization. be used.
- commercially available food-grade emulsifiers for example lecithin, are also suitable. Salts, buffers, sugar, sorbitol, amino acids and others can be present as further additives.
- the overall preparation should be isotonic. To stabilize the fine distribution, microencapsulation of the peptides in question or encapsulation in liposomes can also be provided.
- the peptides to be administered can also be pulverized in the solid state, for example freeze-dried, spray-dried or crystallized from solution, and are then preferably mixed with dry chlorofluorocarbons as propellants and aerosol formers.
- solid additives in particular stabilizers, for example sugar or sugar-like substances, lactose and the like, can be added.
- Inhalation devices are also known in which the aerosol formers or propellants, on the one hand, and the actual pharmaceutical preparation, on the other hand, are stored in different chambers and released together in a predetermined dosage. This avoids inaccurate dosing due to segregation during storage.
- the size of the particles to be inhaled is less critical than in many other applications, since the peptides according to the invention should not be transported into the blood transmembrane, but only have to reach the guanylate cyclase C receptor located apically in the lungs. Particle sizes between 0.5 and 10 ⁇ m appear suitable.
- obstructive and restrictive ventilation disorders These respiratory diseases are characterized by endobronchial obstruction with bronchospasm, edema of the mucous membrane and by hypersecretion of viscous mucus (dyscrine). These phenomena lead to the patient being exhausted by increased and insufficient breathing work. As a restrictive component, gas emissions exchange due to the mucosal edema significantly deteriorated, the oxygen uptake of the lungs significantly reduced.
- the use of the peptides is aimed at an action which counteracts these pathomechanisms.
- the inhalation application relaxes the smooth airway muscles, so that the bronchial resistance and thus the breathing work of the patient decreases. The exhaustion of the patient is reduced or prevented by making breathing work easier.
- the peptides perform different functions, which in their combination and synergy lead to a significant improvement in breathing.
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Abstract
The invention relates to the use of a guanylate cyclase C activated peptide for the treatment of respiratory airway problems and problems associated with ventilation disorder and/or mucous secretion disorders via the airways, in addition to a medicament which is fed via the airways. The invention also relates to an inhalation device which contains the medicament and a method for diagnosing the illnesses associated with inhalation disorders and mucous secretion disorders in the airways, by detecting a gualylate cyclase C activated peptide. The peptides which are used are guanylin, uroguanylin and lymphoguanylin or a heat resistant enterotoxin.
Description
Verwendung eines Peptids, welches Guanylat Cyclase C aktiviert, für die Behandlung von Atemwegserkrankungen über die Luftwege, Arzneimittel, Inhalationsvorrichtung und DiagnoseverfahrenUse of a peptide that activates guanylate cyclase C for the treatment of respiratory diseases via the airways, pharmaceuticals, inhalation device and diagnostic methods
Die Erfindung betrifft die Verwendung eines Peptids, welches Guanylat Cyclase C aktiviert, für die Behandlung von Atemwegserkrankungen und Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, ein zugehöriges Arzneimittel, eine Inhalationsvorrichtung und ein Verfahren zur Diagnose der vorgenannten Erkrankungen.The invention relates to the use of a peptide which activates guanylate cyclase C for the treatment of respiratory diseases and disorders associated with ventilation disorders and / or disorders of mucus secretion, an associated medicament, an inhalation device and a method for diagnosing the aforementioned disorders.
Die obstruktiven Ventilationsstörungen sind ein ernstes klinisches Problem. Sie gehen mit einer Einengung der Atemwege und damit einer Erhöhung des Strömungswiderstands, Spasmen der Bronchialmuskulatur, ödematösen Schwellungen der Bronchialwand sowie gesteigerter Sekretion (Hyperkrinie) von Schleim zäher Konsistenz einher. Die Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, umfassen u.a. Asthma bronchiale, chronische Bronchitis und Mukoviszidose.Obstructive ventilation disorders are a serious clinical problem. They are associated with a narrowing of the airways and thus an increase in flow resistance, spasms of the bronchial muscles, edematous swellings of the bronchial wall and increased secretion (hypercrine) of mucus with a tough consistency. The diseases associated with ventilation disorders and / or disorders of mucus secretion include Bronchial asthma, chronic bronchitis and cystic fibrosis.
Es stehen zur Zeit keine Substanzen zur Verfügung, die nachhaltig und effizient wirksam sind und zur wesentlichen Verbesserung der Symptome führen.There are currently no substances available that have a sustainable and efficient effect and lead to a significant improvement in symptoms.
Als Sekretolytika oder Mukolytika - die auch unter Expektorantien zusammengefasst werden - sind u.a. Bromhexin, Ambroxol, Acetylcystein und Carbocistein im Einsatz. Der therapeutische Wert dieser Substanzen ist jedoch laut Mutschier, "Arzneimittelwirkungen", -Wissenschaftliche Verlagsgesellschaft mbH Stuttgart, 1996, zweifelhaft.As secretolytics or mucolytics - which are also summarized under expectorants - include Bromhexine, ambroxol, acetylcysteine and carbocistein in use. However, the therapeutic value of these substances is doubtful, according to Mutschier, "Medicinal Effects", -Scientific publishing house company Stuttgart Stuttgart, 1996.
Der Erfindung liegt die Aufgabe zugrunde, ein neues effektives Mittel zur Behandlung von Atemwegserkrankungen und allgemein von Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, bereitzustellen, wobei dieses Mittel die Verflüssigung und den besseren Abtransport insbesondere von Bronchialschleim ermöglichen soll.The invention has for its object to provide a new effective agent for the treatment of respiratory diseases and generally diseases that are associated with ventilation disorders and / or disorders of mucus secretion, which agent should allow liquefaction and better removal, especially of bronchial mucus.
Die Aufgabe wird gelöst durch die Verwendung eines Peptids, welches Guanylat Cyclase C aktiviert, für die Herstellung eines Arzneimittels zur Behandlung von Atemwegserkrankungen und Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, über die Luftwege, wobei das Arzneimittel
so formuliert ist, dass die Zuführung des Peptids auf der Luftseite der Atemwege, nämlich zur apikalen Membran der Schleimhaut-Epithelzellen gerichtet, erfolgt.The object is achieved by the use of a peptide which activates guanylate cyclase C for the manufacture of a medicament for the treatment of respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion, via the airways, the medicament is formulated in such a way that the peptide is supplied on the air side of the respiratory tract, namely towards the apical membrane of the mucosal epithelial cells.
Mehrere dieser Peptide können auch gemeinsam oder in Folge verabreicht werden. Äquivalent zur Verwendung dieser Peptide selbst ist die Verwendung homologer, im wesentlichen funktionsgleicher Peptide, insbesondere solcher Peptidvarianten mit durch Deletion, Insertion oder Austausch einzelner und/oder mehrerer Aminosäuren, sequenzverlängerndes Anfügen von einzelnen und/oder mehreren Aminosäuren und/oder chemischer Derivatisierung (insbesondere der terminalen Aminosäuren) verbundener Sequenz-Modifikation.Several of these peptides can also be administered together or in succession. Equivalent to the use of these peptides themselves is the use of homologous, essentially functionally identical peptides, in particular those peptide variants with the addition of single and / or several amino acids by deletion, insertion or exchange of single and / or several amino acids, and / or chemical derivatization (in particular the terminal amino acids) linked sequence modification.
Pharmakologisch verträgliche Derivate sind vorzugsweise amidierte, acetylierte, phosphorylierte und glycosylierte Formen der Peptide und andere posttranslationale Derivatisierungen, einschließlich Salze dieser Peptide und Peptidderivate.Pharmacologically acceptable derivatives are preferably amidated, acetylated, phosphorylated and glycosylated forms of the peptides and other post-translational derivatizations, including salts of these peptides and peptide derivatives.
Es können natürliche, beispielsweise aus Blut, Lymphe, Urin oder humanen oder tierischen Geweben isolierte Peptide oder Peptidgemische, die aufgereinigt seien sollten, oder synthetische oder gentechnisch gewonnene (rekombinante) Peptide eingesetzt werden.Natural peptides or peptide mixtures isolated from blood, lymph, urine or human or animal tissues, which should be purified, or synthetic or genetically engineered (recombinant) peptides can be used.
Bei dem Peptid handelt es sich insbesondere um wenigstens eines der als Guanylin, Uroguanylin und Lymphoguanylin bezeichneten Peptide oder um ein hitzebeständiges Enterotoxin. Diese Peptide sind als solche bekannt. Es kann auch ein zu den genannten Peptiden homologes Peptid mit im wesentlichen gleicher Funktion verwendet werden. Unter den Homologen werden hier solche Peptide verstanden, die weitgehend mit den nachfolgend noch beschriebenen Sequenzen übereinstimmen und vom Fachmann aufgrund ihrer Funktion und Sequenzhomologie noch den Guanylin-Peptiden zugerechnet werden. Dem Fachmann ist bekannt, dass z.B. Punktmutationen, Deletionen und Insertionen die Funktion eines Peptids nicht beeinträchtigen müssen. Derartig veränderte Peptide würden daher zu den Homologen gerechnet.The peptide is in particular at least one of the peptides called guanylin, uroguanylin and lymphoguanylin or a heat-resistant enterotoxin. These peptides are known as such. A peptide homologous to the peptides mentioned and having essentially the same function can also be used. The homologs here are understood to mean those peptides which largely correspond to the sequences described below and are still attributed to the guanylin peptides by the person skilled in the art on the basis of their function and sequence homology. It is known to those skilled in the art that e.g. Point mutations, deletions and insertions do not have to affect the function of a peptide. Peptides modified in this way would therefore be classified as homologues.
Bevorzugt wird derzeit ein Guanylin-Peptid mit 15 Aminosäuren in folgender Sequenz:
Seq. ID 1 (Guanylin, 15 AS): PGTCEICAYAACTGC Pro-Gly-Thr-Cys-Glu-Ile-Cys-Ala-Thr-Ala-Ala-Cys-Thr-Gly-CysA guanylin peptide with 15 amino acids in the following sequence is currently preferred: Seq. ID 1 (Guanylin, 15 AS): PGTCEICAYAACTGC Pro-Gly-Thr-Cys-Glu-Ile-Cys-Ala-Thr-Ala-Ala-Cys-Thr-Gly-Cys
Ein 115 Aminosäuren langes Vorläufermolekül, das die vorstehende Sequenz enthält, wird häufig ebenfalls als "Guanylin" bezeichnet. Beide Peptide sind im Sinne der Erfindung geeignet, bevorzugt ist das Peptid mit Seq. ID 1 , das sich als relativ kleines Peptid gut über die Inhalation zuführen lässt.A 115 amino acid precursor molecule containing the above sequence is also often referred to as "guanylin". Both peptides are suitable for the purposes of the invention; the peptide with Seq is preferred. ID 1, which is a relatively small peptide that can be delivered via inhalation.
Ein 15-AS-Peptid mit der Sequenz PGTCEICAYAACTGC wurde zunächst aus Darmextrakten der Ratte isoliert und als "Guanylin" bezeichnet. Nach der Klonierung und Charakterisierung der cDNA für das menschliche Guanylin war es offensichtlich, dass das Guanylin als Vorläufer-Molekül mit 115 AS (Seq. ID 4: MNAFLLFALC LLGAWAALAG GVTVQDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKP LCKEPNAQEI LQRLEEIAED PGTCEICAYAACTGC) synthetisiert wird. Inzwischen ist bekannt, dass nicht das Vorläufer-Molekül als bioaktives Protein im Blut zirkuliert, sondern das Guanylin mit 94 AS (Proguanylin 22-A 15-AS peptide with the sequence PGTCEICAYAACTGC was first isolated from rat intestinal extracts and referred to as "guanylin". After cloning and characterization of the cDNA for the human Guanylin it was obvious that the Guanylin as a precursor molecule with 115 AS (SEQ ID 4. MNAFLLFALC LLGAWAALAG GVTVQDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKP LCKEPNAQEI LQRLEEIAED PGTCEICAYAACTGC) is synthesized. It is now known that it is not the precursor molecule that circulates in the blood as a bioactive protein, but the guanylin with 94 AS (Proguanylin 22-
115: VTVQDGNFS PGTCEICAYA ACTGC). Der in der Literatur etablierte Begriff115: VTVQDGNFS PGTCEICAYA ACTGC). The term established in literature
"Guanylin" umschreibt sowohl das 15-AS-Peptid als auch das längere 94-AS-Peptid."Guanylin" describes both the 15-AS peptide and the longer 94-AS peptide.
Humanes Uroguanylin ist ein Peptid, dem folgende Aminosäuresequenzen zugeordnet wurde:Human uroguanylin is a peptide to which the following amino acid sequences have been assigned:
Seq. ID 2 (Uroguanylin, 16 AS): NDDCELCVNVACTGCL Asn-Asp-Asp-Cys-Glu-Leu-Cys-Val-Asn-Val-Ala-Cys-Thr-Gly-Cys-LeuSeq. ID 2 (Uroguanylin, 16 AS): NDDCELCVNVACTGCL Asn-Asp-Asp-Cys-Glu-Leu-Cys-Val-Asn-Val-Ala-Cys-Thr-Gly-Cys-Leu
und wurde ursprünglich aus menschlichem Urin isoliert, worauf die Namensgebung beruht. Die US 5 489 670 beschreibt die Isolierung und Synthese von humanem Uroguanylin und sieht eine Verwendung als Laxans gegen Obstipationen vor.and was originally isolated from human urine, which is where the name comes from. US 5,489,670 describes the isolation and synthesis of human uroguanylin and provides use as a laxative against constipation.
Das Uroguanylin wurde zunächst als ein 16-AS-Peptid (NDDC ELCVNVACTG CL) aus dem Harn isoliert. Die Klonierung und Charakterisierung der cDNA für menschliches Uroguanylin ergab ein Uroguanylin Vorläufer-Molekül mit 112 AS (Seq. ID 5: MGCRAASGLL PGVAWLLLL LQSTQSVYIQ YQGFRVQLES MKKLSDLEAQ WAPSPRLQAQSLLPAVCHHP ALPQDLQPVC ASQEASSI FKTLRTIA NDDC ELCVNVACTG CL). Nach Abspaltung des Signalpeptids entsteht ein 86 AS- Uroguanylin (unterstrichene Sequenz). Das 16-AS- und das 86-AS-Peptid werden als Uroguanylin bezeichnet.
Lymphoguanylin ist ein in Lymphgeweben exprimiertes Guanylin-Peptid, das von Forte et al. gefunden wurde (Forte et al. Endocrinology 1999, 140, 1800-1806). Es handelt sich um ein 15 Aminosäuren langes Peptid mit folgender Aminosäuresequenz:The uroguanylin was first isolated from the urine as a 16-AS peptide (NDDC ELCVNVACTG CL). The cloning and characterization of the cDNA for human uroguanylin resulted in a uroguanylin precursor molecule with 112 AS (Seq. ID 5: MGCRAASGLL PGVAWLLLL LQSTQSVYIQ YQGFRVQLES MKKLSDLEAQ WAPSPRLQAQSLLPAVLHVVDCLCLQEQHVVGLCLCQQHVVDCLCLQQHQVVGLCLQQHVVDCLQQQHVVDCLQQQHVVGCLCQLQHVVGQLQQHQVVGLCLQQHVVGQLCQQHVVGGLCLQQHVVGLCL After the signal peptide has been split off, an 86 AS-uroguanylin (underlined sequence) is formed. The 16-AS and 86-AS peptides are called uroguanylin. Lymphoguanylin is a guanylin peptide expressed in lymphoid tissues, which has been described by Forte et al. was found (Forte et al. Endocrinology 1999, 140, 1800-1806). It is a 15 amino acid peptide with the following amino acid sequence:
Seq. ID 3 (Lymphoguanylin, 15 AS): QEECELCINMACTGY Gln-Glu-Glu-Cys-Glu-Leu-Cys-Ile-Asn-Met-Ala-Cys-Thr-Gly-TyrSeq. ID 3 (lymphoguanylin, 15 aa): QEECELCINMACTGY Gln-Glu-Glu-Cys-Glu-Leu-Cys-Ile-Asn-Met-Ala-Cys-Thr-Gly-Tyr
Das Vorläufer-Molekül für Lymphoguanylin umfasst 109 Aminosäuren (Seq. ID 6: MKVLALPMAV TAMLLIL AQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNIFRAL RYIN QEECELCINMACTGY).The precursor molecule for lymphoguanylin contains 109 amino acids (SEQ ID 6: MKVLALPMAV TAMLLIL AQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNEGRELCINM
Die für Lymphoguanylin angegebenen Sequenzen stammen aus dem Opossum. Die menschliche Sequenz ist bisher nicht bekannt. Das 15-Aminosäuren- Lymphoguanylin aktiviert ebenso die menschliche Guanylat Cyclase C.The sequences given for lymphoguanylin come from the opossum. The human sequence is not yet known. The 15-amino acid lymphoguanylin also activates human guanylate cyclase C.
Von den vorgenannten Peptiden ist seit längerer Zeit bekannt, dass sie Guanylat- Cyclase stimulieren oder aktivieren, einen G-Protein-gekoppelten Rezeptor, der die Bildung von zyklischem Guanosinmonophosphat (cGMP) aus Guanosintriphosphat (GTP) katalysiert. Es wurden nacheinander mehrere Guanylat-Cyclase aktivierende Peptide entdeckt, die als endogene Liganden für die Guanylat Cyclase C betrachtet werden. Das erste dieser Peptide wurde Guanylin genannt (Currie, H.G. et al. Proc. Natl. Acad. Sei. USA 1992, 89, 947-951 ).The aforementioned peptides have long been known to stimulate or activate guanylate cyclase, a G protein-coupled receptor that catalyzes the formation of cyclic guanosine monophosphate (cGMP) from guanosine triphosphate (GTP). Several guanylate cyclase activating peptides were discovered in succession, which are considered to be endogenous ligands for guanylate cyclase C. The first of these peptides was named guanylin (Currie, H.G. et al. Proc. Natl. Acad. Sei. USA 1992, 89, 947-951).
Im Darm rufen hitzstabile Enterotoxine - kleine Peptide, die u.a. von pathogenen Escherichia coli Stämmen produziert werden - sekretorische Diarrhöen hervor. Auch diese Toxine entfalten ihre Wirkung durch Stimulation der Guanylat Cyclase C, die von Darmepithellzellen exprimiert wird. Wie die hitzestabilen Enterotoxine führen die Guanylin-Peptide zu einer erhöhten Elektrolyt/Wasser-Sekretion an der Darmschleimhaut. Damit fungiert die Guanylat Cyclase C nicht nur als Rezeptor für die hitzestabilen Enterotoxine, sondern sie stellt den genuinen Rezeptor der endogenen Guanylin-Peptide dar.Heat-stable enterotoxins - small peptides that produced by pathogenic Escherichia coli strains - secretory diarrhea. These toxins also exert their effect by stimulating the guanylate cyclase C, which is expressed by intestinal epithelial cells. Like the heat-stable enterotoxins, the guanylin peptides lead to an increased electrolyte / water secretion on the intestinal mucosa. The guanylate cyclase C thus not only acts as a receptor for the heat-stable enterotoxins, but it also represents the genuine receptor of the endogenous guanylin peptides.
Eine im Rahmen der Erfindung geeignete Sequenz eines hitzestabilen Enterotoxins ist:A sequence of a heat-stable enterotoxin which is suitable in the context of the invention is:
Seq. ID 7 (hitzestabiles Enterotoxin): N S S N Y C C E L C C N P A C T G C Y (19Seq. ID 7 (heat-stable enterotoxin): N S S N Y C C E L C C N P A C T G C Y (19
AS) aus enteropathogenen E. coli.
Gemeinsamer Wirkmechanismus der hitzestabilen Enterotoxine. Guanylin. Uroguanylin und Lymphoguanylin an der Darmschleimhaut.AS) from enteropathogenic E. coli. Common mechanism of action of heat-stable enterotoxins. Guanylin. Uroguanylin and lymphoguanylin on the intestinal mucosa.
In der Darmschleimhaut führen diese oben aufgelisteten Guanylin-Peptide und die hitzestabilen Enterotoxine über die Aktivierung des gemeinsamen Rezeptors zu einem Anstieg von cGMP in den Enterozyten. Durch den erhöhten cGMP-Spiegel wird in den Enterozyten die cGMP-abhängige Proteinkinase II (cGKII) aktiviert. Diese aktivierte Proteinkinase phosphoryliert und öffnet dadurch den CFTR-Chloridkanal in der apikalen Membran der Enterozyten. Dadurch kommt es zu einer Sekretion von Chlorid-Ionen und Wasser in das Lumen des Darms. Der CFTR-Chloridkanal gilt heute als der finale Effektor der Signaltransduktionskette der Guanylin-Peptide. Damit stellen diese Peptide einen direkten Regulator des CFTR-Chlorid-Kanals dar.In the intestinal mucosa, these guanylin peptides listed above and the heat-stable enterotoxins lead to an increase in cGMP in the enterocytes via the activation of the common receptor. The increased cGMP level activates the cGMP-dependent protein kinase II (cGKII) in the enterocytes. This activated protein kinase phosphorylates and thereby opens the CFTR chloride channel in the apical membrane of the enterocytes. This results in the secretion of chloride ions and water into the lumen of the intestine. The CFTR chloride channel is now considered the final effector of the signal transduction chain of the guanylin peptides. These peptides are therefore a direct regulator of the CFTR chloride channel.
Besonderes Augenmerk gilt der Sekretion von Bikarbonat, die auch durch die Guanylin-Peptide vermittelt wird. Nach den bisherigen Erkenntnissen erfolgt die Bikarbonat-Sekretion über einen spezifischen CI7HCO3- - Austauscher (AE-2). Aufgrund bisheriger Befunde kann gefolgert werden, dass das über CFTR luminal se- zernierte CI" wieder in die jeweiligen Zellen aufgenommen und durch HCO3- ausgetauscht wird. Damit kann festgehalten werden, dass die Guanylin-Peptide in den genannten Enterozyten eine zentrale Rolle in der Regulation von CI" und HCO3- spielen. Der Wirkmechanismus der Guanylin-Peptide ist in Abbildung 1 dargestellt.Particular attention is paid to the secretion of bicarbonate, which is also mediated by the guanylin peptides. According to previous knowledge, bicarbonate secretion takes place via a specific CI7HCO 3 - exchanger (AE-2). Based on previous findings, it can be concluded that the CI " luminally secreted via CFTR is taken up again in the respective cells and replaced by HCO 3 -. It can thus be stated that the guanylin peptides play a central role in the enterocytes mentioned Regulation of CI " and HCO 3 - play. The mechanism of action of the guanylin peptides is shown in Figure 1.
Die genannten Peptide zirkulieren als endogene Aktivatoren im Blut. Sie können auch aus Blut bzw. Haemofiltrat gewonnen werden. So wird in der DE 195 28 544 ein Guanylin-Peptid beschrieben, das aus menschlichem Blut gewonnen wurde und für die diagnostische, medizinische und gewerbliche Verwendung als Arzneimittel vorgesehen ist. Dieses Peptid wurde als GCAP-II bezeichnet. Auf Grund der bekannten Wirkung der Guanylin-Peptide auf Guanylat Cylase C (s.o.) wurde GCAP-II speziell für die Behandlung von Erkrankungen, die mit Störungen des Elektrolyttransportes in den Zellen einhergehen vorgesehen. Die Anwendung soll vorzugsweise per Injektion erfolgen.The peptides mentioned circulate in the blood as endogenous activators. They can also be obtained from blood or haemofiltrate. DE 195 28 544 describes a guanylin peptide which was obtained from human blood and is intended for diagnostic, medical and commercial use as a medicament. This peptide was named GCAP-II. Due to the known effect of the guanylin peptides on guanylate cyclase C (see above), GCAP-II was specially designed for the treatment of diseases which are associated with disorders in the electrolyte transport in the cells. The application should preferably be by injection.
Der endogene Aktivator Guanylin wird an verschiedenen Orten im Körper gefunden. Nachgewiesen wurde Guanylin z. B. in der menschlichen Bauchspeicheldrüse (Ku- laksiz et al, Histochem Cell Biol. (2001) 115, 131-145), in der Niere (Forte et al, Annu Rev. Physiol 2000, 62, 673-695) , im Intestinaltrakt (Quian et al, Endocrinology 2000, 141 , 3210-24) und in der Lunge (Cetin et al, Proc. Natl. Acad. Sei. USA, 92, 5925 - 5929, 1995).
Durch die Anmelder konnte nun gefunden werden, dass der gemeinsame Rezeptor für hitzestabile Enterotoxine und Guanylin-Peptide, die Guanylat Cyclase C, in der Schleimhaut der Luftwege lokalisiert ist und dort in hohem Maße auf der apikalen Membran (Luftseite) der jeweiligen Epithelzellen exprimiert wird, nicht jedoch auf der basolateralen Membran (Blutseite). Der in der Lunge lokalisierte Rezeptor kann daher nicht über die Blutbahn, sondern ausschließlich über die Luftwege stimuliert werden.The endogenous activator guanylin is found in various places in the body. Guanylin has been proven e.g. B. in the human pancreas (Kulaksiz et al, Histochem Cell Biol. (2001) 115, 131-145), in the kidney (Forte et al, Annu Rev. Physiol 2000, 62, 673-695), in the intestinal tract (Quian et al, Endocrinology 2000, 141, 3210-24) and in the lungs (Cetin et al, Proc. Natl. Acad. Sci. USA, 92, 5925 - 5929, 1995). It has now been found by the applicants that the common receptor for heat-stable enterotoxins and guanylin peptides, the guanylate cyclase C, is located in the mucosa of the airways and is expressed there to a high degree on the apical membrane (air side) of the respective epithelial cells. but not on the basolateral membrane (blood side). The receptor located in the lungs can therefore not be stimulated via the bloodstream, but only via the airways.
Der Wirkungsmechanismus auf zellulärer und molekularer Ebene wird in der Figur 1 dargestellt, die schematisch die Signaltransduktion der Guanylin-Peptide an Epithelzellen zeigt.The mechanism of action at the cellular and molecular level is shown in FIG. 1, which shows schematically the signal transduction of the guanylin peptides on epithelial cells.
Guanlyat Cyclase (GC-C) ist ein Enzym-Rezeptor-Komplex, der als Membranprotein ausschließlich in der apikalen, zur Atemwege-Lichtung hin gerichteten Zelldomäne lokalisiert ist. Er fehlt an der basolateralen Membran der Zellen (Blutseite), die bekanntlich in Kontakt mit dem zirkulierenden Blut steht.Guanlyate cyclase (GC-C) is an enzyme-receptor complex that is localized as a membrane protein exclusively in the apical cell domain, which is directed towards the airway clearing. It lacks the basolateral membrane of the cells (blood side), which is known to be in contact with the circulating blood.
Guanylin-Peptide, die über die Lichtung der Atemwege an den Rezeptor (GC-C) binden, setzen einen spezifischen intrazellulären Mechanismus in Gang, der verschiedene Proteinmodule enthält. Die durch die Guanylin-Peptide von außen aktivierte GC-C bildet intrazellulär in hohen Mengen cGMP aus GTP. Dieser second messenger (cGMP) aktiviert eine membranassoziierte cGMP-abhängige Proteinkinase Typ II (cKGII), die die Phosphorylierung und damit Aktivierung des CFTR-Proteins an seiner regulatorischen (R-) Domäne vornimmt. CFTR ist ein Membranprotein in der apikalen Membran der Epithelzellen und ist ein wichtiger Chlorid-Kanal, der nach Aktivierung Chlorid-Ionen aus der Zelle in Richtung Lichtung der Atemwege sezerniert. Aufgrund des so entstandenen ionischen Gradienten folgt das Wasser den sezer- nierten Chlordi-Ionen und fließt in die Lichtung der Atemwege. Das Wasser stammt aus den Epithelzellen und aus den Zwischenräumen zwischen den Zellen (parazellulär). Ein Teil der in die Lichtung sezernierten Chlorid-Ionen wird erneut in die Zellen aufgenommen; dafür werden Bikarbonat-Ionen aus den Zellen sezerniert. Dieser Austauch von Ionen wird durch den Anionen-Austauscher Typ II (AE2) bewerkstelligt. Auch das AE2-Protein ist in der apikalen Membran der Epithelzellen lokalisiert. Intrazellulär werden die Bikarbonat-Ionen durch das Enzym Carboanhydrase Typ II (CAM) aus Wasser und Kohlendioxid hergestellt.Guanylin peptides that bind to the receptor (GC-C) via the airway clearing set in motion a specific intracellular mechanism that contains various protein modules. The GC-C activated externally by the guanylin peptides forms cGMP from GTP in high amounts intracellularly. This second messenger (cGMP) activates a membrane-associated cGMP-dependent protein kinase type II (cKGII), which carries out the phosphorylation and thus activation of the CFTR protein on its regulatory (R) domain. CFTR is a membrane protein in the apical membrane of the epithelial cells and is an important chloride channel that, after activation, secretes chloride ions from the cell towards the airway clearing. Because of the ionic gradient created in this way, the water follows the secreted chlorine ions and flows into the clearing of the respiratory tract. The water comes from the epithelial cells and from the spaces between the cells (paracellular). Some of the chloride ions secreted into the clearing are taken up again in the cells; for this, bicarbonate ions are secreted from the cells. This exchange of ions is accomplished by the Type II (AE2) anion exchanger. The AE2 protein is also located in the apical membrane of the epithelial cells. The bicarbonate ions are produced intracellularly by the enzyme carbonic anhydrase type II (CAM) from water and carbon dioxide.
Damit ist die luftseitige Membran der Epithelzellen der Schleimhaut die entscheidende Stelle der Signal-Rezeption, regulatorischen Aktivität und Elektrolyt/Wasser- sezernierenden Kapazität in den Atemwegen.
Insgesamt werden aufgrund dieses Wirkmechanismus der Guanylin-Peptide Ionen und Flüssigkeit in die Lichtung der Atemwege sezerniert, die die Qualität und Fließeigenschaften des Bronchialschleims maßgeblich beeinflussen und bestimmen.This makes the airside membrane of the epithelial cells of the mucous membrane the crucial point for signal reception, regulatory activity and electrolyte / water-secreting capacity in the respiratory tract. Overall, due to this mechanism of action of the guanylin peptides, ions and liquid are secreted into the airway clearing, which significantly influence and determine the quality and flow properties of the bronchial mucus.
In der Figur werden folgende Abkürzungen verwendet: GC-C = Guanylat Cyclase C; cGKM = cGMP-abhängige Proteinkinase Typ II; CFTR = cystic fibrosis transmembra- ne conductance regulator; AE-2 = Anionenaustauscher Typ 2; CAM = Carboanhydrase Typ II.The following abbreviations are used in the figure: GC-C = guanylate cyclase C; cGKM = cGMP-dependent protein kinase type II; CFTR = cystic fibrosis transmembrane conductance regulator; AE-2 = anion exchanger type 2; CAM = carbonic anhydrase type II.
Die Aufklärung des der Erfindung zugrundeliegenden Wirkmechanismus wurde veröffentlicht in "Kulaksiz, H., Schmid, A., Hönscheid, M., Ramaswamy, A., Cetin, Y., PNAS, May 2002, Vol. 99, Seiten 6796-6801", "Kulaksiz et al., Histochem Cell Biol. (2001 115,131-145",The elucidation of the mechanism of action on which the invention is based was published in "Kulaksiz, H., Schmid, A., Hönscheid, M., Ramaswamy, A., Cetin, Y., PNAS, May 2002, Vol. 99, pages 6796-6801" , "Kulaksiz et al., Histochem Cell Biol. (2001 115, 131-145",
Eine zentrale Erkenntnis des erfindungsgemäßen Konzepts ist, dass die Aktivierung des Rezeptors durch Applikation der endogenen Liganden gezielt über die Luftwege zu erfolgen hat. Der Fachmann muss daher die Zuführung des Peptids oder des Arzneimittels, das das Peptid enthält, so einstellen, dass das Peptid - möglichst ausschließlich - auf der Luftseite zur apikalen Membran der Atemwege zugeführt wird und nicht etwa in größerem Ausmaß in die Blutbahn gelangt. Gerade hierdurch wird die gezielte lokale therapeutische Anwendung im Atemtrakt ermöglicht, zumal der Rezeptor in den Atemwegen ausschließlich luftseitig lokalisiert ist.A central finding of the concept according to the invention is that the activation of the receptor by application of the endogenous ligands has to be carried out specifically via the airways. The person skilled in the art must therefore adjust the supply of the peptide or the medicament which contains the peptide in such a way that the peptide is fed - if possible exclusively - on the air side to the apical membrane of the respiratory tract and does not reach the bloodstream to a greater extent. This enables targeted local therapeutic use in the respiratory tract, especially since the receptor in the respiratory tract is located exclusively on the air side.
Bei der Zuführung der erfindungsgemäßen Peptide, nämlich der Guanylat Cyclase C- Liganden über die Luftwege handelt es sich um eine gerichtete und unmittelbare Zuführung zu dem luftseitig gelegenen Rezeptor. Eine Erhöhung der Blutkonzentration des Peptids durch Aufnahme über die Lunge, wie bei der Inhalation andere Peptide (die systemisch wirden, z.B. Insulin) angestrebt, soll hier gerade strikt vermieden werden.The supply of the peptides according to the invention, namely the guanylate cyclase C ligand via the airways, is a directed and direct supply to the receptor located on the air side. An increase in the blood concentration of the peptide by ingestion via the lungs, as is the case with inhalation other peptides (which are systemic, e.g. insulin), should be strictly avoided here.
Dem Fachmann stehen hierfür die geeigneten Mittel zur Verfügung. Er kann die gerichtete Zuführung zur Luftseite über die Einstellung der Peptidkonzentration in der Arzneimittelformulierung, die Dosierung und die Einstellung der Partikel/Tröpfchengröße innerhalb der Formulierung oder des Inhalationsmittels so beeinflussen, dass praktisch kein Peptid zur Blutseite der Atemwege (zur basolateralen Membran) und damit in die Blutbahn durchtritt. Die optimalen Bedingungen können für jedes gewählte Peptid in gezielten Vorversuchen ermittelt werden.
Die Erfindung ermöglicht eine Therapie mit Dosen, die sehr viel geringer sind als solche, die für die Erhöhung der Blutkonzentration erforderlich wären, unter Minimierung bis Ausschaltung der systemischen Nebenwirkungen der jeweiligen Peptide.The appropriate means are available to the person skilled in the art for this. It can influence the directed supply to the air side via the adjustment of the peptide concentration in the pharmaceutical formulation, the dosage and the adjustment of the particle / droplet size within the formulation or the inhalation agent in such a way that practically no peptide on the blood side of the respiratory tract (to the basolateral membrane) and thus in enters the bloodstream. The optimal conditions for each selected peptide can be determined in targeted preliminary tests. The invention enables therapy with doses that are much lower than those that would be required to increase the blood concentration, while minimizing or eliminating the systemic side effects of the respective peptides.
Nur bei einer Applikation über die Luft führen die hitzestabilen Enterotoxine und die genannten Guanylin-Peptide zu einer ausreichenden Aktivierung des Rezeptors Guanylat Cyclase C und dadurch zu einer erhöhten Flüssigkeitssekretion in den Atemwegen. Bei einer systemischen Applikation wäre außerdem mit unerwünschten Nebenreaktionen zu rechnen, beispielsweise führt das Enterotoxin zu sehr unangenehmen sekretorischen Durchfallerkrankungen.The heat-stable enterotoxins and the guanylin peptides mentioned only lead to sufficient activation of the guanylate cyclase C receptor and thus to increased fluid secretion in the respiratory tract only when administered via the air. With a systemic application, undesirable side reactions would also be expected, for example the enterotoxin leads to very unpleasant secretory diarrheal diseases.
Weiterhin wirken die erfindungsgemäßen Peptide als Stimulantien im Sinne einer Sekretolyse durch Auflösen des in den Luftwegen vorliegenden zähen Schleims, wobei die Ionen-Zusammensetzung und der pH-Wert der Flüssigkeit unmittelbar auf den Epithelzellen ("Mikroklima") so eingestellt werden, dass der zähe Schleim sich zunehmend "verflüssigt".Furthermore, the peptides according to the invention act as stimulants in the sense of secretolysis by dissolving the viscous mucus present in the airways, the ion composition and the pH of the liquid being adjusted directly on the epithelial cells (“microclimate”) in such a way that the viscous mucus increasingly "liquefied".
Der Abtransport von Schleim und Mikropartikeln aus den Atemwegen wird durch Epithelzellen ermöglicht, die auf ihrer apikalen Seite (Luftseite) Flimmerhärchen (Zilien) tragen. Die "reinigende" Funktion wird durch Schlagen (rachenwärts) der Zilien erreicht.The removal of mucus and microparticles from the respiratory tract is made possible by epithelial cells that have cilia on their apical side (air side). The "cleaning" function is achieved by beating the cilia.
Da die Guanylin-Peptide nebst ihrer Funktion, die Elektrolyt- und Wasser-Sekretion zu erhöhen, insbesondere auch die Zilien-tragenden Epithelzellen aktivieren, kommt es an diesen Zellen zu einer erhöhten Schlagfrequenz der Zilien. Damit wird im Sinne einer konzertierten Aktion das Sekret und kleinste Partikel auf der Schleimhaut der Atemwege wesentlich effizienter abtransportiert, was die physiologische und therapeutische Bedeutung der Guanylin-Peptide unterstreicht.Since the guanylin peptides, in addition to their function of increasing the electrolyte and water secretion, in particular also activate the cilia-bearing epithelial cells, these cells have an increased beat frequency of the cilia. In the sense of a concerted action, the secretions and tiny particles on the mucous membrane of the respiratory tract are removed much more efficiently, which underlines the physiological and therapeutic importance of the guanylin peptides.
Weiterhin ist anzuführen, dass die genannten Substanzen relaxierend auf die glatte Muskulatur in der Wand der Bronchien und Bronchioli wirken. Dies führt insgesamt zu einer wesentlich verbesserten Atmung.It should also be mentioned that the substances mentioned have a relaxing effect on the smooth muscles in the wall of the bronchi and bronchioli. Overall, this leads to significantly improved breathing.
Die vorgenannten neugefundenen Eigenschaften der erfindungsgemäßen Peptide wirken synergistisch im Sinne der Erfindung zusammen und führen zu der sehr guten Wirkung der durch die Luftwegen zugeführten Peptide zur Behandlung der eingangs genannten Störung und Erkrankungen.
Die erfindungsgemäßen Peptide können auf Basis dieser Erkenntnisse zusätzlich für die Herstellung von Diagnostika für Atemwegserkrankungen und Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, verwendet werden.The aforementioned newly found properties of the peptides according to the invention act synergistically in the sense of the invention and lead to the very good effect of the peptides supplied by the airways for the treatment of the disorder and diseases mentioned at the beginning. On the basis of these findings, the peptides according to the invention can additionally be used for the production of diagnostics for respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion.
Zunächst sind hierfür die Peptide selbst als Referenzsubstanzen für die Diagnostik geeignet. Ein Fehlen/Mangel oder ein Überschuss dieser Peptide beispielsweise in Bronchialschleim, Exsudat oder Lavage kann das Vorhandensein behandlungsbedürftiger Störungen anzeigen. Der Nachweis der Peptide kann mit den üblichen und bekannten Mitteln, wie spektrokopisch, chromatographisch oder chemisch geschehen.First of all, the peptides themselves are suitable as reference substances for diagnostics. A lack / deficiency or an excess of these peptides, for example in bronchial mucus, exudate or lavage, can indicate the presence of disorders requiring treatment. The peptides can be detected using the customary and known means, such as spectrocopically, chromatographically or chemically.
Weiterhin können für diesen Nachweis vom Fachmann mit Hilfe dafür üblicher Verfahren und Mittel Antikörper gegen die erfindungsgemäßen Peptide hergestellt werden, die dann innerhalb molekularbiologischer bzw. enzymatischer Assays eingesetzt werden können.For this detection, the person skilled in the art can use conventional methods and means to produce antibodies against the peptides according to the invention, which can then be used within molecular biological or enzymatic assays.
Zur Lösung der Aufgabe der Erfindung trägt daher auch ein Verfahren zur Diagnose der genannten Erkrankungen bei, bei welchem wenigstens eines der Peptide, das Guanylat Cyclase C aktiviert, nachgewiesen wird, und zwar vorzugsweise im Bronchialschleim, Exsudat, Lavage, Nasensekret oder Speichel.To achieve the object of the invention therefore also contributes to a method for diagnosing the diseases mentioned, in which at least one of the peptides that activate guanylate cyclase C is detected, preferably in bronchial mucus, exudate, lavage, nasal secretions or saliva.
Der Nachweis kann durch Nachweis einer der Sequenzen zu Seq. ID 1 bis ID 6The detection can be carried out by detecting one of the sequences for Seq. ID 1 to ID 6
Seq. ID 1 (Guanylin): PGTCEICAYA ACTGCSeq. ID 1 (Guanylin): PGTCEICAYA ACTGC
Seq. ID 4 (Guanylin-Vorläufer-Molekül): MNAFLLFALC LLGAWAALAG GV QDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKPLCKEPNAQEI LQRLEEIAED PGTCEICAYA ACTGCSeq. ID 4 (Guanylin precursor molecule): MNAFLLFALC LLGAWAALAG GV QDGNFS FSLESVKKLK DLQEPQEPRV GKLRNFAPIP GEPWPILCS NPNFPEELKPLCKEPNAQEI LQRLEEIAED PGTCEICAYA ACTGC
Seq. ID 2 (Uroguanylin): NDDC ELCVNVACTGCL Seq. ID 5 (Uroguanylin-Vorläufer-Molekül): MGCRAASGLLPGVAWLLLL LQSTQSVYIQ YQGFRVQLES MKKLSDLEAQ WAPSPRLQAQ SLLPAVCHHPALPQDLQPVC ASQEASSIFK TLRTIAN DDC ELCVNVACTG CLSeq. ID 2 (Uroguanylin): NDDC ELCVNVACTGCL Seq. ID 5 (Uroguanylin precursor molecule): MGCRAASGLLPGVAWLLLL LQSTQSVYIQ YQGFRVQLES MKKLSDLEAQ WAPSPRLQAQ SLLPAVCHHPALPQDLQPVC ASQEASSIFK TLRTIAN DDC ELCVNVACTG
Seq. ID 3 (Lymphoguanylin): QEECELCINMACTGYSeq. ID 3 (lymphoguanylin): QEECELCINMACTGY
Seq. ID 6 (Lymphoguanylin-Vorläufer-Molekül): MKVLALPMAVTAMLLILAQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNIFRAL RYIN QEECELCINMACTGY
Seq. ID 7 (hitzestabiles Enterotoxin): N S S N Y C C E L C C N P A C T G C Y (19 AS) aus enteropathogenen E. coli.Seq. ID 6 (lymphoguanylin precursor molecule): MKVLALPMAVTAMLLILAQN TQSVYIQYEG FQVNLDSVKK LDKLLEQLRG FHHQMGDQRD PSILCSDPALPSDLQPVCEN SQAVNIFRAL RYIN QEECELCINM Seq. ID 7 (heat-stable enterotoxin): NSSNYCCELCCNPACTGCY (19 AS) from enteropathogenic E. coli.
erfolgen. Als positives Testergebnis für den Nachweis einer Störung wird gewertet, wenn eine von Vergleichsproben gesunder Probanden abweichende Konzentration wenigstens eines der Peptide, die Guanylat Cyclase C aktivieren, gefunden wird.respectively. A positive test result for the detection of a disorder is evaluated if a concentration of at least one of the peptides that activate guanylate cyclase C that deviates from comparison samples of healthy volunteers is found.
Die erfindungsgemäße Verwendung der Peptide besteht weiter darin, dass ein Arzneimittel formuliert wird, welches über die Luftwege zugeführt wird und wenigstens ein Peptid enthält, das Guanylat Cyclase C aktiviert. Diese Peptide wurden oben bereits ausführlich beschrieben.The use of the peptides according to the invention further consists in formulating a medicament which is supplied via the airways and contains at least one peptide which activates guanylate cyclase C. These peptides have already been described in detail above.
Neben dem Peptid oder dem Peptidgemisch kann wenigstens ein weiterer Wirkstoff sowie gegebenenfalls Hilfs- und Zusatzstoffe in dem Arnzeimittel enthalten sein. Als weitere Wirkstoffe kommen hier beispielsweise muskelrelaxierende Mitteln, Lokalantibiotika, vorwiegend für die Behandlung gleichzeitig aufgepfropfter bakterieller Infektionen, oder auch zusätzliche Mukolytika, Sekretolytike, Antitussiva oder bronchodi- latierende Substanzen in Betracht. Die Auswahl wird der Fachmann auf Basis der jeweiligen Bedürfnisse bei der Behandlung der eingangs genannten Erkrankungen treffen.In addition to the peptide or the peptide mixture, at least one further active ingredient and optionally auxiliaries and additives can be present in the drug. Other active ingredients here are, for example, muscle relaxants, local antibiotics, primarily for the treatment of simultaneously grafted bacterial infections, or also additional mucolytics, secretolytics, antitussives or bronchodilating substances. The specialist will make the selection on the basis of the respective needs in the treatment of the diseases mentioned at the beginning.
Das Arzneimittel kann in fester oder flüssiger Form zubereitet werden und wird vom Benutzer in geeigneter Weise über die Luftwege zugeführt. Hierfür kann es mit einem handelsüblichen Zerstäuber oder Inhalationsgerät verabreicht werden.The drug can be prepared in solid or liquid form and is conveniently supplied by the user via the airways. For this purpose it can be administered with a commercially available nebulizer or inhalation device.
In bevorzugter Ausführungsform liegt das Arzneimittel als Inhalationsmittel vor und enthält wenigstens ein Treibmittel. Als Treibmittel eignen sich besonders Fluorchlorkohlenwasserstoffe. Geeignete Treibmittel sind dem Fachmann auf diesem Gebiet bekannt. Allgemein können alle geeigneten Aerosolbildner oder auch Rauchbildner verwendet werden. Je nach Hilfsstoff wird ein Aerosol oder ein Rauch inhaliert, wobei ein Aerosol bevorzugt ist.In a preferred embodiment, the medicament is present as an inhalation agent and contains at least one propellant. Chlorofluorocarbons are particularly suitable as blowing agents. Suitable blowing agents are known to those skilled in the art. In general, all suitable aerosol formers or smoke formers can be used. Depending on the excipient, an aerosol or smoke is inhaled, with an aerosol being preferred.
Zur Lösung der Aufgabe ist schließlich eine Inhalationsvorrichtung vorgesehen, die das Arzneimittel enthält, d.h. dass das Arzneimittel in der Inhalationsvorrichtung fertig konfektioniert vorliegt. Eine solche Inhalationsvorrichtung kann aus einer Sprühvorrichtung, insbesondere einer Dosier-Sprühvorrichtung oder einem Dosier-Inhalator (englisch: MDI, metered dose inhaler) bestehen. Geeignete Inhalatoren sind dem Fachmann bekannt und beispielsweise beschrieben in US 3 915 165, EP 166476 und US 6 099 517. Geeignet sind auch Ultraschallvernebler.
Die erfindungsgemäßen Peptide sollten für die Verabreichung zunächst in eine feindisperse Form überführt werden. Hierfür können sie zunächst in Lösung oder Suspension gebracht und gegebenenfalls mit pharmazeutischen verträglichen Zusätzen in dieser Form stabilisiert werden. Zur Stabilisierung können verträgliche Tenside, z.B. Tween ®. verwendet werden. Geeignet sind je nach Inhalationsverfahren auch handelsübliche als Lebensmittel zugelassene Emulgatoren, z.B. Lecithin. Als weitere Zusatzstoffe können Salze, Puffer, Zucker, Sorbitol, Aminosäuren u.a.m. vorhanden sein. Die Gesamtzubereitung sollte isotonisch sein. Zur Stabilisierung der Feinverteilung kann ebenfalls eine Mikroverkapselung der betreffenden Peptide oder eine Verkapselung in Liposome vorgesehen sein.Finally, to achieve the object, an inhalation device is provided which contains the medicament, ie that the medicament is present in the inhalation device ready-made. Such an inhalation device can consist of a spray device, in particular a metering spray device or a metering inhaler (English: MDI, metered dose inhaler). Suitable inhalers are known to the person skilled in the art and are described, for example, in US Pat. No. 3,915,165, EP 166476 and US Pat. No. 6,099,517. Ultrasonic nebulizers are also suitable. The peptides according to the invention should first be converted into a finely dispersed form for administration. For this purpose, they can first be brought into solution or suspension and, if necessary, stabilized in this form with pharmaceutically acceptable additives. Compatible surfactants such as Tween ® can be used for stabilization. be used. Depending on the inhalation process, commercially available food-grade emulsifiers, for example lecithin, are also suitable. Salts, buffers, sugar, sorbitol, amino acids and others can be present as further additives. The overall preparation should be isotonic. To stabilize the fine distribution, microencapsulation of the peptides in question or encapsulation in liposomes can also be provided.
Die zu verabreichenden Peptide können auch im festen Zustand pulverisiert, beispielsweise aus Lösung gefriergetrocknet, sprühgetrocknet oder kristallisiert, vorliegen und werden dann bevorzugt mit trockenen Fluorchlorkohlenwasserstoffen als Treibmittel und Aerosolbildner gemischt. Bei pulverförmiger Verabreichung können feste Zusätze, insbesondere Stabilisatoren, beispielsweise Zucker oder zuckerartige Stoffe, Lactose und dergleichen, zugesetzt sein.The peptides to be administered can also be pulverized in the solid state, for example freeze-dried, spray-dried or crystallized from solution, and are then preferably mixed with dry chlorofluorocarbons as propellants and aerosol formers. In the case of powdery administration, solid additives, in particular stabilizers, for example sugar or sugar-like substances, lactose and the like, can be added.
Es sind auch Inhalationsvorrichtungen bekannt, in denen die Aerosolbildner oder Treibmittel einerseits und die eigentliche Arzneimittelzubereitung andererseits in verschiedenen Kammern aufbewahrt und gemeinsam in vorgegebener Dosierung abgegeben werden. Dies vermeidet ungenaue Dosierung durch Entmischung bei Lagerung.Inhalation devices are also known in which the aerosol formers or propellants, on the one hand, and the actual pharmaceutical preparation, on the other hand, are stored in different chambers and released together in a predetermined dosage. This avoids inaccurate dosing due to segregation during storage.
Die Größe der zu inhalierenden Partikel ist weniger kritisch als bei vielen anderen Anwendungen, da die erfindungsgemäßen Peptide nicht transmembran ins Blut transportiert werden sollen, sondern lediglich den in der Lunge apikal lokalisierten Rezeptor Guanylat Cyclase C erreichen müssen. Teilchengrößen zwischen 0,5 und 10 μm erscheinen geeignet.The size of the particles to be inhaled is less critical than in many other applications, since the peptides according to the invention should not be transported into the blood transmembrane, but only have to reach the guanylate cyclase C receptor located apically in the lungs. Particle sizes between 0.5 and 10 μm appear suitable.
Im folgenden wird die Erfindung anhand eines Beispiels erläutert:The invention is explained below using an example:
Die Anwendung der Peptide soll am Beispiel der "obstruktiven und restriktiven Ventilationsstörungen" erläutert werden. Diese Atemwegserkrankungen sind gekennzeichnet durch eine endobronchiale Obstruktion mit Bronchospasmus, Schleimhautödem und durch eine Hypersekretion eines zähen Schleims (Dyskrinie). Diese Erscheinungen führen dazu, dass der betroffene Patient durch vermehrte und insuffizi- ente Atemarbeit regelrecht erschöpft. Als restriktive Komponente wird der Gasaus-
tausch durch das Schleimhautödem wesentlich verschlechtert, die Sauerstoffaufnahme der Lungen deutlich vermindert.The use of the peptides will be explained using the example of "obstructive and restrictive ventilation disorders". These respiratory diseases are characterized by endobronchial obstruction with bronchospasm, edema of the mucous membrane and by hypersecretion of viscous mucus (dyscrine). These phenomena lead to the patient being exhausted by increased and insufficient breathing work. As a restrictive component, gas emissions exchange due to the mucosal edema significantly deteriorated, the oxygen uptake of the lungs significantly reduced.
Die Anwendung der Peptide zielt auf eine diesen Pathomechanismen entgegenstehende Wirkung ab. Die inhalative Applikation führt zu einer Relaxierung der glatten Atemwegs-Muskulatur, so dass der bronchiale Widerstand und damit die Atemarbeit des Patienten abnimmt. Mit der Erleichterung der Atemarbeit wird eine Erschöpfung des Patienten gemindert bzw. verhindert.The use of the peptides is aimed at an action which counteracts these pathomechanisms. The inhalation application relaxes the smooth airway muscles, so that the bronchial resistance and thus the breathing work of the patient decreases. The exhaustion of the patient is reduced or prevented by making breathing work easier.
Aufgrund von Elektrolyt/Wasser-sezemierenden Wirkungen dieser Peptide wird eine vermehrte Wasserausschwemmung aus der Schleimhaut der Atemwege induziert, die im Sinne einer Abnahme des Schleimhautödems (Abschwellung) wirkt und damit zu einer verbesserten Atmung führt. Durch den vermehrten Wasseraustritt aus der Schleimhaut wird die Dyskrinie vermindert, der zähe Schleim verflüssigt und der Abtransport des Sekretes durch erhöhten Zilienschlag verbessert.Due to the electrolyte / water-secreting effects of these peptides, an increased outflow of water from the mucous membrane of the respiratory tract is induced, which works in the sense of a decrease in the mucosal edema (swelling) and thus leads to improved breathing. Due to the increased water leakage from the mucous membrane, the dyscrine is reduced, the viscous mucus is liquefied and the discharge of the secretion is improved by increased ciliary beat.
Somit üben die Peptide unterschiedliche Funktionen aus, die in ihrer Kombination und Synergie zu einer deutlichen Verbesserung der Atmung führen.
Thus, the peptides perform different functions, which in their combination and synergy lead to a significant improvement in breathing.
Claims
1. Verwendung eines Peptids, welches Guanylat Cyclase C aktiviert, für die Herstellung eines Arzneimittels zur Behandlung von Atemwegserkrankungen und Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen, über die Luftwege, wobei das Arzneimittel so formuliert ist, dass die Zuführung des Peptids auf der Luftseite der Atemwege, nämlich zur apikalen Membran der Schleimhaut-Epithelzellen gerichtet, erfolgt.1. Use of a peptide which activates guanylate cyclase C for the manufacture of a medicament for the treatment of respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion, via the airways, the medicament being formulated in such a way that the supply of the Peptide on the air side of the airways, namely directed towards the apical membrane of the mucosal epithelial cells.
2. Verwendung nach Anspruch 1 , dadurch gekennzeichnet, dass das Peptid ein natürliches oder rekombinantes Guanylin, Uroguanylin, Lymphoguanylin oder hitzebeständiges Enterotoxin ist, oder ein zu diesen homologes, im wesentlichen funktionsgleiches Peptid, insbesondere eine solche Peptidvariante mit durch Deletion, Insertion oder Austausch einzelner und/oder mehrerer Aminosäuren, sequenzverlängerndes Anfügen von einzelnen und/oder mehreren Aminosäuren und/oder chemischer Derivatisierung insbesondere der terminalen Aminosäuren verbundener Sequenz- Modifikation.2. Use according to claim 1, characterized in that the peptide is a natural or recombinant guanylin, uroguanylin, lymphoguanylin or heat-resistant enterotoxin, or a peptide which is homologous and essentially has the same function, in particular such a peptide variant with by deletion, insertion or exchange of individual and / or several amino acids, sequence-extending addition of single and / or several amino acids and / or chemical derivatization, in particular of the terminal amino acids linked sequence modification.
3. Verwendung nach Anspruch 1 oder 2, dadurch gekennzeichnet, dass das Peptid eine der Sequenzen zu Seq. ID 1 bis Seq. ID 7 umfasst.3. Use according to claim 1 or 2, characterized in that the peptide is one of the sequences to Seq. ID 1 to Seq. ID 7 includes.
4. Verwendung eines Peptids, wie in einem der Ansprüche 1 bis 3 angegeben, für die Herstellung eines Diagnostikums für Atemwegserkrankungen und Erkrankungen, die mit Ventilationsstörungen und/oder Störungen der Schleimsekretion einhergehen.4. Use of a peptide, as specified in one of claims 1 to 3, for the production of a diagnostic for respiratory diseases and diseases which are associated with ventilation disorders and / or disorders of mucus secretion.
5. Arzneimittel in einer Zubereitung, welche über die Luftwege an der apikalen Membran zugeführt wird, dadurch gekennzeichnet, dass es wenigstens ein Peptid enthält, das Guanylat Cyclase C aktiviert.5. Medicament in a preparation which is supplied to the apical membrane via the airways, characterized in that it contains at least one peptide which activates guanylate cyclase C.
6. Arzneimittel nach Anspruch 5, dadurch gekennzeichnet, dass das Peptid Guanylin, Uroguanylin, Lymphoguanylin oder ein hitzebeständiges Enterotoxin ist, oder ein zu diesen homologes, im wesentlichen funktionsgleiches Peptid, insbesondere ein solcher Peptidvarianten mit durch Deletion, Insertion oder Austausch einzelner und/oder mehrerer Aminosäuren, sequenzverlängerndes Anfügen von einzelnen und/oder mehreren Aminosäuren und/oder chemischer Derivatisierung insbesondere der terminalen Aminosäuren verbundener Sequenz-Modifikation, oder ein wenigstens eines dieser Peptide enthaltendes Peptidgemisch.6. Medicament according to claim 5, characterized in that the peptide is guanylin, uroguanylin, lymphoguanylin or a heat-resistant enterotoxin, or a homologous, essentially functionally identical peptide, in particular such a peptide variant with by deletion, insertion or exchange of individual and / or several amino acids, sequence-extending addition of individual and / or several amino acids and / or chemical derivatization, in particular of the terminal amino acids linked sequence modification, or a peptide mixture containing at least one of these peptides.
7. Arzneimittel nach Anspruch 5 oder 6, dadurch gekennzeichnet, dass wenigstens eines der Peptide eine der Sequenzen zu Seq. ID 1 bis Seq. ID 7 umfasst.7. Medicament according to claim 5 or 6, characterized in that at least one of the peptides one of the sequences to Seq. ID 1 to Seq. ID 7 includes.
8. Arzneimittel nach einem der Ansprüche 5 bis 7, dadurch gekennzeichnet, dass das Arzneimittel neben dem wenigstens einen Peptid als Wirkstoff wenigstens einen weiteren Wirkstoff enthält, sowie gegebenenfalls Hilfs- und Zusatzstoffe.8. Medicament according to one of claims 5 to 7, characterized in that the medicament contains in addition to the at least one peptide as an active ingredient at least one further active ingredient, and optionally auxiliaries and additives.
9. Arzneimittel nach einem der Ansprüche 5 bis 8, dadurch gekennzeichnet, dass das Arzneimittel in Form eines Inhalationsmittels vorliegt und wenigstens ein Treibmittel, wenigstens einen Aerosolbildner oder wenigstens einen Rauchbildner enthält.9. Medicament according to one of claims 5 to 8, characterized in that the medicament is in the form of an inhalation agent and contains at least one propellant, at least one aerosol generator or at least one smoke generator.
10. Inhalationsvorrichtung, enthaltend das Arzneimittel nach einem der Ansprüche 5 bis 9.10. inhalation device containing the medicament according to any one of claims 5 to 9.
11. Inhalationsvorrichtung nach Anspruch 10, dadurch gekennzeichnet, dass sie eine Sprühvorrichtung, insbesondere eine Dosier-Sprühvorrichtung oder einen Dosier- Inhalator umfasst.11. Inhalation device according to claim 10, characterized in that it comprises a spray device, in particular a metering spray device or a metering inhaler.
12. Verfahren zur Diagnose von Erkrankungen, die mit Ventilationsstörungen und Störungen der Schleimsekretion in den Atemwegen einhergehen, durch Nachweis wenigstens eines Peptids, das Guanylat Cyclase C aktiviert.12. A method for the diagnosis of diseases which are associated with ventilation disorders and disorders of mucus secretion in the respiratory tract, by detecting at least one peptide which activates guanylate cyclase C.
13. Verfahren nach Anspruch 12, dadurch gekennzeichnet, dass der Nachweis auf wenigstens eine der Sequenzen zu Seq. ID 1 bis Seq. ID 7 gerichtet ist.13. The method according to claim 12, characterized in that the detection of at least one of the sequences to Seq. ID 1 to Seq. ID 7 is directed.
14. Verfahren nach Anspruch 12 oder 13, dadurch gekennzeichnet, dass das Peptid in Exsudat, Bronchialschleim oder Lavage nachgewiesen wird.14. The method according to claim 12 or 13, characterized in that the peptide is detected in exudate, bronchial mucus or lavage.
15. Verfahren nach einem der Ansprüche 12 bis 14, dadurch gekennzeichnet, dass eine von Vergleichsproben gesunder Probanden abweichende Konzentration wenig- stens eines der Peptide, die Guanylat Cyclase C aktivieren, als positives Testergebnis für den Nachweis einer Störung gewertet wird. 15. The method according to any one of claims 12 to 14, characterized in that a concentration deviating from comparison samples of healthy test persons At least one of the peptides that activate guanylate cyclase C is considered a positive test result for the detection of a disorder.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE10127119 | 2001-06-05 | ||
| DE10127119 | 2001-06-05 | ||
| PCT/DE2002/002040 WO2002098912A2 (en) | 2001-06-05 | 2002-06-05 | Guanylate-cyclase c ligand, administered via the airways, for the treatment of respiratory airway problems |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| EP1392729A2 true EP1392729A2 (en) | 2004-03-03 |
Family
ID=7687161
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EP02745124A Withdrawn EP1392729A2 (en) | 2001-06-05 | 2002-06-05 | Use of a peptide which activates guanylate-cyclase c for the treatment of respiratory airway problems via the airways, medicament, inhalation devices and method of diagnosis |
Country Status (6)
| Country | Link |
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| US (1) | US20050032684A1 (en) |
| EP (1) | EP1392729A2 (en) |
| AU (1) | AU2002316771A1 (en) |
| CA (1) | CA2464511A1 (en) |
| DE (1) | DE10292434D2 (en) |
| WO (1) | WO2002098912A2 (en) |
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| CA2810243C (en) | 2010-09-15 | 2021-04-20 | Synergy Pharmaceuticals Inc. | Formulations of guanylate cyclase c agonists and methods of use |
| US9708367B2 (en) | 2013-03-15 | 2017-07-18 | Synergy Pharmaceuticals, Inc. | Agonists of guanylate cyclase and their uses |
| CN114404588A (en) | 2013-08-09 | 2022-04-29 | 阿德利克斯公司 | Compounds and methods for inhibiting phosphate transport |
| EP3972599B1 (en) | 2019-05-21 | 2025-10-22 | Ardelyx, Inc. | Combination for lowering serum phosphate in a patient |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE19543628A1 (en) * | 1995-11-24 | 1997-05-28 | Forssmann Wolf Georg | Human circulating peptide with insulinotropic activity (GCAP-II- (89-112), (guanylyl cyclase C activating peptide II) and its GCAP analogues, in particular GCAP-I- (99-115), its use as a pharmacological Active ingredient and use of its principle of action to provide new GC-C-dependent insulinotropic active ingredients |
-
2002
- 2002-06-05 EP EP02745124A patent/EP1392729A2/en not_active Withdrawn
- 2002-06-05 WO PCT/DE2002/002040 patent/WO2002098912A2/en not_active Ceased
- 2002-06-05 DE DE10292434T patent/DE10292434D2/en not_active Expired - Fee Related
- 2002-06-05 AU AU2002316771A patent/AU2002316771A1/en not_active Abandoned
- 2002-06-05 US US10/479,606 patent/US20050032684A1/en not_active Abandoned
- 2002-06-05 CA CA002464511A patent/CA2464511A1/en not_active Abandoned
Non-Patent Citations (1)
| Title |
|---|
| See references of WO02098912A2 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO2002098912A3 (en) | 2003-07-31 |
| US20050032684A1 (en) | 2005-02-10 |
| AU2002316771A1 (en) | 2002-12-16 |
| DE10292434D2 (en) | 2004-07-29 |
| CA2464511A1 (en) | 2002-12-12 |
| WO2002098912A2 (en) | 2002-12-12 |
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