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    • 专利摘要:
    A new family of compounds, method of obtaining them and their uses in the treatment and/or prophylaxis of diseases. The present invention describes compounds of formula (I) and pharmaceutical compositions containing it. It also describes a process for obtaining said compounds of formula (I). Finally, the present invention deals with the use of said compounds and of the pharmaceutical compositions containing them, for preparing a medicament. Especially drugs intended for the treatment and/or prophylaxis of diseases derived from the anomalous transport of anions at cellular level, and more specifically to those intended for the treatment and/or prophylaxis of cystic fibrosis. (Machine-translation by Google Translate, not legally binding)
    公开号:ES2615077A1
    申请号:ES201531749
    申请日:2015-12-02
    公开日:2017-06-05
    发明作者:Roberto QUESADA PATO；Elsa HERNANDO SANTA CRUZ；Ricardo Enrique PÉREZ TOMÁS；Vanessa SOTO CERRATO；Olga L.A. ZEGARRA
    申请人:Universitat Autonoma de Barcelona UAB；Universitat de Barcelona UB；Universidad de Burgos；
    IPC主号:
    专利说明:

    A new family of compounds, procedure for obtaining them and their uses in the treatment and / or prophylaxis of diseases

    DESCRIPTION
     5
    FIELD OF THE INVENTION
    The field of the invention is related to small molecules capable of facilitating the transport of anions through lipid membranes. These molecules are able to replace the lost function in diseases related to deficiencies in the transport of anions at the cellular level. The compounds of the invention are particularly useful in the treatment of diseases such as cystic fibrosis.

    BACKGROUND OF THE INVENTION
    The compounds of the invention have not been described in the state of the art. There are related compounds, such as prodigiosins, natural pigments initially isolated from bacteria of the genus Serratia. These naturally occurring alkaloids contain the 4-methoxy-2,2'-bipyrrole unit and show cytotoxic and immunosuppressive activities (Fürstner, A. Chemistry and Biology of Roseophilin and the Prodigiosin Alkaloids: A Survey of the Last 2500 Years. Angew. Chem Int. Ed. 2003, 42, 3582-3603). Other compounds are also known in the state of the art, such as obatoclax, a member of the family of 20 prodiginins with anticancer properties, which is currently in clinical trials (Nguyen, M. et al. Small molecule obatoclax (GX15 -070) MCL-1 and overcomes MCL-1-mediated resistance to apoptosis antagonists. PNAS, 2007, 104, 19512-19517). A general characteristic of these compounds is the fact that they are cytotoxic even at very low concentrations. On the other hand, it has been shown that some of these compounds facilitate the transport of anions in lipid membranes and this action has been related to their cytotoxicity (Díaz de Greñu, B. et al. Synthetic Prodiginine Obatoclax (GX15-070) and Related Analogues : Anion Binding, Transmembrane Transport, and Cytotoxicity Properties. Chem. Eur. J., 2011, 17, 14074-14083). There are numerous diseases related to dysfunctions of membrane proteins 30 responsible for facilitating the transport of ions through biological membranes or ion channels. These diseases are very diverse and are grouped into what is known as “channelopathies” (F. M. Ashcroft, F. M. From molecule to malady. Nature, 2006, 440, 440–447). The development of molecules or compounds capable of supplying the activity of these ion channels, which is altered in the case of these diseases, would be therapeutically desirable. Cystic fibrosis (CF) is a genetic disease that gives
    This results in a severe reduction in the function of a protein called cystic fibrosis transmembrane conductance regulator (CFTR) that regulates the permeability of anions in the epithelial membranes. This condition results in an imbalance in ion transport and a reduced apical secretion of anions. As a consequence, an accumulation of abnormally dense mucus in the lung occurs and this results in recurrent microbial infections that damage lung function and can ultimately lead to death in patients with cystic fibrosis. In addition, these patients with cystic fibrosis usually suffer from gastrointestinal problems and pancreatic insufficiency. Currently there is no known cure for this disease. In addition to palliative treatments of the 10 symptoms, a corrector for the treatment of cystic fibrosis called kalydeco (ivacaftor) and a combination, called Orkambi (ivacaftor + lumacaftor), of a corrector plus an enhancer is approved in the market. In both cases, these treatments are aimed at patients with specific mutations and are not of general application for all CF patients. The mechanism of action of these compounds 15 always involves an action on the CFTR protein in such a way that the activity of the protein present in the membrane (enhancer effect) is enhanced or defects that occur in the transit of this protein to the protein are corrected. the plasma membrane in which its action develops (corrective effect). The development of drugs capable of supplying the transport function of the CFTR protein would be a promising therapeutic agent to correct the reduced apical conductance of these cells affected by this disease. In the state of the art, no treatment using this strategy to achieve therapeutic action has been described. In addition, unlike current treatments, these compounds would be able to act on the origin of the disease, being potentially applicable to all cystic fibrosis patients 25 regardless of the mutation causing it.

    DESCRIPTION OF THE INVENTION
    The present invention relates to the compound of formula (I)

    where R1, R2, R3, R6, R7 and R8 are independently selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl,
    R4 and R5 are selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, 5 COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl,
    Wherein R9, R10 and R11 are radicals independently selected from the group consisting of: H, OH, halogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl.
    In the present invention, the term "alkyl" refers to aliphatic, linear or branched chains, among which are, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, sec-butyl, pentyl , isopentyl and dodecyl.
    In the present invention, the term "cycloalkyl" refers to cycles of 3 to 6 members, among which are, but are not limited to, cyclobutyl, cyclopentyl and cyclohexyl.
    In the present invention, the term "alkenyl" refers to hydrocarbon groups containing double bonds, among which are but not limited to 2- butenyl, 3-pentenyl and 4-octenyl.
    In the present invention, the term "alkynyl" refers to groups containing triple bonds, among which are, but are not limited to, 2-butynyl and 2-hexinyl.
    In the present invention, the term "aryl" refers to aromatic mono and polycyclic compounds, among which are, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, diphenyl, indenyl, phenanthryl and anthracil.
    In the present invention, the term "arylalkyl" refers to an aliphatic chain in which at least one of the hydrogens has been replaced by an aryl group, as defined above, among those found, but not limited to , benzyl and propylphenyl. 25
    In the present invention, the term "heterocycle" refers to cyclic compounds of 4 to 6 members containing 1 to 4 heteroatoms such as oxygen, sulfur, nitrogen, among which are, but not limited to, pyrrole, pyridyl, pyridazyl, triazolyl , isoxazolidinyl, furyl, thiophenyl and thiazolyl.
    In the present invention, the term "heteroaryl" is a five to six membered aromatic heteroaryl, monocyclic, or a bicyclic seven to eleven aromatic heteroaryl, which in each case contains 1, 2, 3 or 4 heteroatoms selected, independently from each other, from the group consisting of N, S or O
     5
    An embodiment is the compound of the invention, wherein the compound of formula (I) is selected from the group consisting of:
    (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3 chloride -methoxy-2H-pyrrolium (Ia), (Z) Chloride -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H- pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) Chloride -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3- methoxy-2-10 ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl) chloride -4-pentyl-1H-pyrrole-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id).

    Another embodiment of the invention is the preferred synthetic route for the preparation of the compounds of the invention and which is illustrated in the following scheme:


    The process is carried out in two stages from a 5-alkynyl-2H-pyrrole-2-ylidene compound of formula (III). Reaction of this type of compounds of formula (III) with an organic azide R8N3 results in the formation of a 5- (1,2,3-triazol-4-yl) -1H-pyrrole-2-carbaldehyde of formula 20 (II). The condensation of this type of compounds of formula (II) with an unsubstituted pyrrole in the alpha position of formula (IV), in the presence of catalytic amounts of an organic or inorganic acid leads to obtaining the compounds of the invention of formula ( I).

    Therefore, this embodiment of the invention consists in a process for obtaining the compound of formula (I), which comprises:
    (a) the cycloaddition reaction of an R8N3 azide and a 5-alkynyl-2H-pyrrol-2-ylidene compound of formula (III) preferably using a copper catalyst to generate a 5- (1,2,3-triazole-) 4-yl) -1H-pyrrole-2-carbaldehyde of formula (II).
    (b) This compound of formula (II) is condensed with an unsubstituted pyrrole in the alpha position of formula (IV) in the presence of a catalytic amount of acid and an organic solvent.
    (c) The organic solvent is evaporated and the compound of formula (I) desired is obtained by chromatographic column purification. 5
    In the present invention, copper mixtures of copper (II) sulphate and sodium ascorbate are acceptable copper catalysts for carrying out the compound of the invention.
    In the present invention they are acceptable acids for the realization of obtaining the compound of the invention hydrochloric acid and trifluoroacetic acid. 10
    As organic solvents, methanol is preferably used.

    The compounds of the invention of formula (I) and their pharmaceutically acceptable addition salts and / or solvates are particularly useful as chemotherapeutic agents for the treatment of cystic fibrosis and other diseases derived from the anomalous transport of anions at the cellular level.

    Pharmaceutically acceptable addition salts and / or solvates, hydrochloride salts or mesylate are preferred in the present invention.
     twenty
    An embodiment of the present invention is the intermediate of the cycloaddition reaction (a) described in the process of obtaining the compounds of formula (I), characterized by being a triazole-pyrrole-carbaldehyde of formula (II):

     25
    where R4 and R5 are selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl,
    R6, R7 and R8 are independently selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl. 30
    Wherein R9, R10 and R11 are radicals independently selected from the group consisting of: H, OH, halogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl.

    An embodiment of the present invention is the intermediate of the cycloaddition reaction (a) described in the process of obtaining the compounds of formula (I), represented by 5- (1-butyl-1H-1,2, 3-Triazol-4-yl) -3-methoxy-1H-pyrrole-2-carbaldehyde of formula (IIa):

       10
    Another embodiment of the present invention is the intermediate of the cycloaddition reaction (a) described in the process of obtaining the compounds of formula (I), represented by 5- (1-octyl-1H-1,2,3 -triazol-4-yl) -3-methoxy-1H-pyrrole-2-carbaldehyde of formula (IIb):
     fifteen
    An embodiment of the present invention is a pharmaceutical composition comprising a compound of formula (I)

    where R1, R2, R3, R6, R7 and R8 are independently selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl.
    R4 and R5 are selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl. 5
    Wherein R9, R10 and R11 are radicals independently selected from the group consisting of: H, OH, halogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl, together with pharmaceutically acceptable excipients.

    Another embodiment of the present invention is a pharmaceutical composition comprising a compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-) chloride triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrole-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-) chloride octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), chloride of (Z) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl ) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-15 4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-) chloride octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id) together with pharmaceutically acceptable excipients.

    The pharmaceutical composition of the present invention comprising a compound of formula (I) is administered by one of the routes of administration selected from the group consisting of: oral, intravenous, muscular and intramuscular route.

    Another particular embodiment of the present invention is a pharmaceutical composition comprising a compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-) chloride triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrole-2-25 yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1) chloride -octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-) chloride il) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- (1-) chloride octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id) and which is administered by one of the administration routes selected from the group consisting of oral, intravenous, muscular and intramuscular route .

    An embodiment of the invention is the use of the compound of formula (I) to prepare a medicament.
     35
    Another embodiment of the invention is the use of a pharmaceutical composition comprising the compound of formula (I) to prepare a medicament.

    Another embodiment of the invention is the compound of formula (I) for use as a medicament.
     5
    Another embodiment of the present invention is a pharmaceutical composition comprising the compound of formula (I) for use as a medicament.

    An embodiment of the invention is the use of the compound of formula (I) independently selected from the group consisting of: ((Z) -5- (1-butyl-1H-1,2,3-10 triazole-4 chloride) -yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H chloride) -1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) chloride ) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-) chloride 1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament.

    Another embodiment of the invention is the use of a pharmaceutical composition comprising the compound of formula (I) independently selected from the group consisting of: ((Z) -5- (1-butyl-1H-1,2 chloride), 3-Triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-20 2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib ), (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-) chloride 2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament.
     25
    An embodiment of the invention is the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) chloride - 2 - ((3,5-Dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1,2 chloride) , 3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- chloride (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H-30 pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-1,2) chloride , 3-triazol-4-yl) -2H-pyrrolium (Id), for use as a medicine.

    Another embodiment of the invention is a pharmaceutical composition comprising the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-triazole chloride) -4-yl) -2 - ((3,5-dimethyl-1H-pyrrole-2-yl) methylene) -3-methoxy-
    2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrole-2) chloride -yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 chloride - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl) chloride -1H-pyrrole-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), for use as a medicine. 5

    An embodiment of the present invention is the use of the compound of formula (I) to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.
     10
    Diseases derived from the anomalous transport of anions at the cellular level in the context of this invention are cystic fibrosis, Best's disease, Bartter's syndrome, chronic obstructive pulmonary disease or congenital mitonia.

    Another embodiment of the present invention is the use of a pharmaceutical composition comprising the compound of formula (I) to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    An embodiment of the present invention is the compound of formula (I) for use in the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    Another embodiment of the present invention is a pharmaceutical composition comprising the compound of formula (I) for use in the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    An embodiment of the present invention is the use of the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-triazol-4- chloride) il) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), 30 (Z) -5- (1-octyl-1H chloride) -1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) chloride ) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-) chloride 1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    Another embodiment of the present invention is the use of a pharmaceutical composition containing the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2 chloride), 3-Triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-5 dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib ), (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-) chloride 2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level. 10

    An embodiment of the present invention is the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) chloride -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1) chloride, 2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), 15 (Z) chloride - 5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1 H -pyrrol-2-yl) methylene) -2H -pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-1, 2,3-triazol-4-yl) -2H-pyrrolium (Id), for use in the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.
     twenty
    Another embodiment of the present invention is the pharmaceutical composition comprising the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-triazole chloride) -4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl) chloride -1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), chloride (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2-25 ((5-methyl-4-pentyl-1H-pyrrole-2-yl ) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl) chloride -1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), for use in the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.
     30
    An embodiment of the present invention is the use of the compound of formula (I) to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.

    Another embodiment of the present invention is the use of a pharmaceutical composition comprising the compound of formula (I) to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.

    An embodiment of the present invention is the compound of formula (I) for use in the treatment and / or prophylaxis of cystic fibrosis.

    Another embodiment of the present invention is a pharmaceutical composition comprising the compound of formula (I) for use in the treatment and / or prophylaxis of cystic fibrosis.
    An embodiment of the present invention is the use of the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2,3-triazol-4- chloride) il) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-) chloride 1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium 10 (Ib), (Z) chloride ) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-) chloride 1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.
     fifteen
    An embodiment of the present invention is the use of a pharmaceutical composition comprising the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-butyl-1H-1,2 chloride), 3-Triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib) , (Z) -5- (1-Butyl-1H-1,2,3-20 triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-) chloride 2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- ( 1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.
     25
    Another embodiment of the present invention is the compound of formula (I) independently selected from the group consisting of: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) chloride -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1) chloride, 2,3-Triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5 chloride - (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H- 30 pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-1, 2,3-triazol-4-yl) -2H-pyrrolium (Id), for use in the treatment and / or prophylaxis of cystic fibrosis.

    Another embodiment of the present invention is a pharmaceutical composition comprising the compound of formula (I) independently selected from the group formed
    by: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) chloride -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-) chloride 1H-pyrrole-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-butyl-1H-1,2,3-triazol-4-yl) chloride - 3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 chloride ((5- methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id), for use in the 5 treatment and / or prophylaxis of cystic fibrosis.

    An embodiment of the present invention is a method of treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level, which comprises administering to a human subject a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I ).

    Another embodiment of the present invention is a method of treatment and / or prophylaxis of cystic fibrosis, which comprises administering to a human subject a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I).

    An embodiment of the present invention is a method of treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level, which comprises administering to a human subject a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I ) independently selected from the group consisting of: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-) chloride pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2- chloride ((3,5-Dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-butyl-1H-1,2,3) chloride -triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -2H-pyrrolium (Ic), chloride of (Z) -3 -methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2H- pyrrolium (Id).

    Another embodiment of the present invention is a method of treatment and / or prophylaxis of cystic fibrosis, which comprises administering to a subject a pharmaceutical composition comprising a therapeutically effective amount of the compound of formula (I) independently selected from the group formed by: (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) chloride -3-methoxy-2H-pyrrolium (Ia), (Z) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-) chloride 1H-pyrrole-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-butyl-1H-1,2,3-triazol-4-yl) chloride - 3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-35
    il) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- (1-) chloride octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id).

    An embodiment of the present invention is a method of treatment and / or prophylaxis of cystic fibrosis comprising administering the compound of formula (I), in a concentration of 0.1 µM to 50 µM, preferably in a concentration of 15 µM at 40 µM and more preferably, when it comes to combining greater therapeutic efficacy by minimizing the toxicity of the drug administered, when the concentration of the compound of formula (I) to be administered is in the range of 0.1 µM to 20 µM.
      10
    BRIEF DESCRIPTION OF THE FIGURES

    Figure 1. Fluorescence assay used to measure the activity of the compounds. Example of the test used to determine the activity of the compounds. The panel shows the response of cells incubated for 40 minutes with compound Ic at various concentrations and in triplicate. The application of NaI induces a reduction in fluorescence, the speed of which depends on the concentration of the compound: 2 µM (light gray), 8.3 µM (dark gray) and 20 µM (black).

    Figure 2. Dose-response of compounds Ia, Ib, Ic and Id. The maximum rate of fluorescence extinction (VmEF) is obtained from experiments such as the example shown in Figure 1.

    Figure 3. pH dependence of the activity of the compounds Ia, Ib, Ic and Id. The figure shows the maximum fluorescence extinction rate (VmEF) of the four compounds (concentration: 20 µM) at the pH values indicated. As a comparison, the response to two pH values of the CFTR protein is also shown, whose activity is stimulated by the application of an agonist that increases intracellular cAMP (20 µM forskolin) and an enhancer (10 µM genistein).
     twenty
    Figure 4. Kinetics of incorporation of the enhancer compound Ic (12 µM) into the cell membrane. In the continuous presence of the compound in the extracellular solution, the activity of Ic in the cell membrane, measured as VmFE, increases with time.

    Figure 5. Stability of compound Ic (12 µM) in the cell membrane. After incubation for 25 minutes, compound Ic is washed from the extracellular solution to establish its stability in the cell membrane. We see that the activity of Ic in the membrane decreases very slowly, leaving more than 85% of the transport after 60 minutes.
     30

    PREFERRED EMBODIMENTS

    Example 1. Synthesis of intermediate compounds IIa and IIb.

     5

    5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-1H-pyrrole-2-carbaldehyde (IIa). To a mixture of the alkyne III (204 mg, 1 mmol) and 1-butanoazide (99 mg, 1 mmol) dissolved in tBuOH (4 mL) and water (2 mL) are added CuSO4 (0.2 equiv., Dissolved in 2 mL of water), K2CO3 (1 equiv.) and sodium ascorbate (0.4 equiv.). The resulting mixture is stirred vigorously for 3 days at room temperature. Subsequently, the reaction mixture is diluted with 50 mL of NH4OH, and after 10 minutes it is extracted with Et2O (3 x 25 mL). The organic phase is washed with a saturated NaCl solution (1 x 40 mL), dried over Na2SO4 and concentrated under reduced pressure. The solid obtained is recrystallized from a mixture DCM: hexane: Et2O to obtain compound IIa. Yield: 67%. 1H NMR (300 MHz, CDCl3): δ = 10.76 (s, 1H), 9.51 (s, 15 1H), 8.26 (s, 1H), 6.37 (s, 1H), 4.42 (t, J = 7.2 Hz, 2H ), 3.92 (s, 3H, OCH3), 1.99–1.87 (m, 2H), 1.47–1.33 (m, 2H), 0.98 (t, J = 7.4 Hz, 3H); 13C NMR (75 MHz, CDCl3): δ = 174.44 (CHO), 159.79 (C), 140.01 (C), 131.72 (C), 121.25 (CH), 119.10 (C), 93.55 (CH), 58.23 (OCH3) , 50.42 (CH2), 32.31 (CH2), 19.80 (CH2), 13.58 (CH3); HRMS (EI) m / z [M] + calculated for [C12H16N4O2] 248.1273; Found: 248.1282. twenty


    5- (1-Octyl-1H-1,2,3-triazol-4-yl) -3-methoxy-1H-pyrrole-2-carbaldehyde (IIb). To a mixture of the alkyne III (204 mg, 1 mmol) and 1-octanoazide (155 mg, 1 mmol) dissolved in tBuOH (4 mL) and water (2 mL) are added CuSO4 (0.2 equiv., Dissolved in 2 mL of water), K2CO3 (1 equiv.) and 25 sodium ascorbate (0.4 equiv.). The resulting mixture is stirred vigorously for 3 days at room temperature. Subsequently the reaction mixture is diluted with 50 mL of
    NH4OH, and after 10 minutes is extracted with Et2O (3 x 25 mL). The organic phase is washed with a saturated NaCl solution (1 x 40 mL), dried over Na2SO4 and concentrated under reduced pressure. The solid obtained is recrystallized from a mixture DCM: hexane: Et2O to obtain compound IIb. Yield: 76%. 1H NMR (300 MHz, CDCl3): δ = 11.21 (s, 1H), 9.49 (s, 1H), 8.43 (s, 1H), 6.44 (s, 1H), 4.40 (t, J = 6.3 Hz, 2H) , 3.92 (s, 3H, OCH3), 2.09–1.78 (m, 5 2H), 1.47–1.11 (m, 10H), 0.86 (t, J = 6.4 Hz, 3H); 13C NMR (75 MHz, CDCl3): δ = 174.31 (CHO), 160.04 (C), 140.02 (C), 132.12 (C), 121.42 (CH), 119.13 (C), 93.56 (CH), 58.19 (OCH3) , 50.69 (CH2), 31.83 (CH2), 30.34 (CH2), 29.17 (CH2), 29.08 (CH2), 26.57 (CH2), 22.71 (CH2), 14.18 (CH3). HRMS (EI) m / z [M] + calculated for [C16H24N4O2] 304.1899;
     10
    Example 2: Synthesis of compound Ia

    (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3 chloride -methoxy-2H-pyrrolium (Ia). A mixture of carbaldehyde IIa (1 equiv, 0.3 mmol) and 2,4-dimethylpyrrole (2 equiv, 0.6 mmol) is dissolved in 5 mL of MeOH under a nitrogen atmosphere at room temperature. To this is added HCl dissolved in MeOH (2 equiv, 0.6 mmol) dropwise in 15 minutes. Once a TLC plate shows that the starting material has been consumed the solvent is evaporated under reduced pressure. The resulting residue is purified by chromatographic column in aluminum oxide (hexane / AcOEt 3: 1) to obtain compound Ia. 66% yield. 1H NMR (300 MHz, CDCl3): δ = 20 13.55 (s, 1H), 13.24 (s, 1H), 9.69 (s, 1H), 7.24 (s, 1H), 6.66 (d, J = 1.3 Hz, 1H ), 6.12 (s, 1H), 4.44 (t, J = 7.2 Hz, 2H), 4.02 (s, 3H, OCH3), 2.60 (s, 3H), 2.32 (s, 3H), 2.0–1.91 (m, 2H), 1.44–1.32 (m, 2H), 0.95 (t, J = 7.3 Hz, 3H). 13C NMR (75 MHz, CDCl3): δ = 165.61 (C), 153.52 (C), 145.58 (C), 144.73 (C), 138.54 (C), 126.60 (CH), 126.43 (C), 119.16 (C) , 117.19 (CH), 116.91 (CH), 94.58 (CH), 58.99 (OCH3), 50.60 (CH2), 32.17 (CH2), 19.77 (CH2), 25 14.41 (CH3), 13.54 (CH3), 12.17 (CH3 ). HRMS (EI) m / z [M] + calculated for [C18H23N5O] 325.1903; Found: 325.1913.

    Example 3: Synthesis of compound Ib represented by the formula:
    Ib
    (Z) -5- (1-Octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrole-2-yl) methylene) -3 chloride -methoxy-2H-pyrrolium (Ib). A mixture of carbaldehyde IIb (1 equiv, 0.3 mmol) and 2,4-dimethylpyrrole (2 equiv, 0.6 mmol) is dissolved in 5 mL of MeOH under a nitrogen atmosphere at room temperature. To this, HCl dissolved in MeOH (2 equiv, 0.6 mmol) is added dropwise in 15 minutes. Once a TLC plate shows that the starting material has been consumed the solvent is evaporated under reduced pressure. The resulting residue is purified by chromatographic column on aluminum oxide (hexane / AcOEt 3: 1) to obtain compound Ib. 60% yield. 1H NMR (300 MHz, CDCl3): δ = 13.58 (s, 1H), 13.29 (s, 1H), 9.69 (s, 1H), 7.26 (s, 1H), 6.69 (s, 1H), 6.13 (s, 1H), 4.43 (t, J = 10 7.3 Hz, 2H), 4.04 (s, 3H, OCH3), 2.62 (s, 3H), 2.34 (s, 3H), 2.03–1.89 (m, 2H), 1.42– 1.16 (m, 10H), 0.86 (t, J = 6.6 Hz, 3H). 13C NMR (75 MHz, CDCl3): δ = 165.65 (C), 153.55 (C), 145.67 (C), 144.74 (C), 138.59 (C), 126.62 (CH), 126.48 (C), 119.23 (C) , 117.24 (CH), 116.94 (CH), 94.64 (CH), 59.02 (OCH3), 50.97 (CH2), 31.84 (CH2), 30.34 (CH2), 29.18 (CH2), 29.08 (CH2), 26.58 (CH2) , 22.73 (CH2), 14.46 (CH3), 14.21 (CH3), 12.20 (CH3). HRMS (EI) m / z [M] + calculated for [C22H31N5O] 381.2529; Found: 381.2524.
    Example 4: Synthesis of compound Ic represented by the formula:
    Ic
    (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-) chloride il) methylene) -2H-pyrrolium (Ic). twenty
    A mixture of carbaldehyde IIa (1 equiv, 0.3 mmol) and 2-methyl-3-pentylpyrrole (2 equiv, 0.6 mmol) is dissolved in 5 mL of MeOH under a nitrogen atmosphere at room temperature. To this is added HCl dissolved in MeOH (2 equiv, 0.6 mmol) dropwise in 15 minutes. Once a TLC plate shows that the starting material has been consumed the solvent is evaporated under reduced pressure. The resulting residue is purified by chromatographic column in aluminum oxide (hexane / AcOEt 3: 1) to obtain compound Ic. Yield: 57%. 1H NMR (300 MHz, CDCl3): δ = 13.77 (s, 1H), 13.29 (s, 1H), 9.70 (s, 1H), 7.18 (s, 1H), 6.82 (d, J = 2.0 Hz, 1H) , 6.68 (d, J = 1.8 Hz, 1H), 4.45 (t, J = 7.3 Hz, 2H), 4.03 (s, 3H, OCH3), 2.60 (s, 3H), 2.40 (t, J = 7.6 Hz, 2H), 2.02–1.91 (m, 2H), 1.60–1.49 (m, 2H), 1.44–1.28 (m, 6H), 0.97 (t, J = 7.4 Hz, 3H), 0.89 (t, J = 6.9 Hz , 3H). 13C 10 NMR (75 MHz, CDCl3): δ = 165.76 (C), 152.78 (C), 145.75 (C), 138.43 (C), 131.51 (CH), 131.07 (C), 126.61 (CH), 126.46 (C ), 119.91 (CH), 119.68 (C), 94.57 (CH), 58.99 (OCH3), 50.56 (CH2), 32.11 (CH2), 31.43 (CH2), 29.48 (CH2), 25.36 (CH2), 22.51 (CH2 ), 19.72 (CH2), 14.07 (CH3), 13.49 (CH3), 12.81 (CH3). HRMS (EI) m / z [M] + calculated for [C22H31N5O] 381.2529; Found: 381.2524. fifteen

    Example 5: Synthesis of compound Id represented by the formula:
    Id
    (Z) -3-Methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrol-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazole-) chloride 4-yl) -2H-pyrrolium (Id). A mixture of carbaldehyde IIb (1 equiv, 0.3 mmol) and 2-methyl-20 3-pentylpyrrole (2 equiv, 0.6 mmol) is dissolved in 5 mL of MeOH under a nitrogen atmosphere at room temperature. To this is added HCl dissolved in MeOH (2 equiv, 0.6 mmol) dropwise in 15 minutes. Once a TLC plate shows that the starting material has been consumed the solvent is evaporated under reduced pressure. The resulting residue is purified by chromatographic column on aluminum oxide (hexane / AcOEt 25 3: 1) to obtain compound Ib. Yield 63%. 1H NMR (300 MHz, CDCl3): δ = 13.73 (s, 1H), 13.26 (s, 1H), 9.66 (s, 1H), 7.14 (s, 1H), 6.79 (s, 1H), 6.64 (s, 1H), 4.41 (t, J = 7.3 Hz, 2H), 3.99 (s, 3H, OCH3), 2.56 (s, 3H), 2.37 (t, J = 7.6 Hz, 2H), 2.02–1.87 (m, 2H ), 1.59–1.46 (m, 2H), 1.35–1.15 (m, 14H), 0.85 (dt, J = 6.8, 6.9 Hz, 6H). 13C NMR (75 MHz,
    CDCl3): δ = 165.84 (C), 152.86 (C), 145.86 (C), 138.52 (C), 131.57 (CH), 131.15 (C), 126.65 (CH), 126.53 (C), 120.00 (CH), 119.77 (C), 94.66 (CH), 59.05 (OCH3), 50.96 (CH2), 31.82 (CH2), 31.50 (CH2), 30.31 (CH2), 29.57 (CH2), 29.17 (CH2), 29.06 (CH2), 26.57 (CH2), 25.44 (CH2), 22.71 (CH2), 22.59 (CH2), 14.19 (CH3), 14.15 (CH3), 12.90 (CH3). HRMS (EI) m / z [M] + calculated for [C26H39N5O] 437.3155; Found: 437.3159. 5

    Example 6. Toxicity of compounds Ia, Ib, Ic and Id in different cell lines. The following cell lines were used:
    - MCF-7: human breast adenocarcinoma cells.
    - A549: human lung adenocarcinoma cells. 10
    - MCF-10A: normal mammary human epithelium cells.
    The effect of derivatives Ia, Ib, Ic and Id on the viability of different cancer cell lines (MCF-7 and A459) and non-cancer cell lines (MCF-10A) was determined by the MTT assay, where MTT means bromide of 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium, blue thiazole. In this test, 10x103 cells were seeded in plates of 15 96-well culture and after 24 hours they were incubated in the absence (control cells) or in the presence of different concentrations of Ia, Ib, Ic and Id for 24 hours. Subsequently, 10 µM of MTT was added for 4 hours and the absorbances were measured in a spectrophotometer at a length of 570 nm. These tests allowed calculating the concentrations necessary to decrease 50% of cell viability. This value, IC50, 20 is shown in the following table.
     Compound  IC50 (µM) MCF-7 IC50 (µM) A549 IC50 (µM) MCF-10A
     Ia  > 100 92.70 ± 3.54 79.92 ± 9.74
     Ib  > 100> 100> 100
     Ic  46.62 ± 7.35 34.99 ± 11.49 31.16 ± 1.58
     Id  33.86 ± 16.12 54.32 ± 15.45 32.96 ± 5.99

    The results indicate a low to very moderate cytotoxicity of these compounds. All cell anion transport assays have been performed at concentrations below those indicated, such that the viability of the cells is not compromised under those conditions.

    Example 7
    Functional measures of the ability to transport anions of compounds Ia, Ib, Ic and Id. 30
    Cells:
    The FRT (Fisher rat thyroid) cell line, a rat thyroid line with stable expression of the yellow fluorescent protein (YFP), was used. The cells are seeded in 96-well plates at a density of 40,000 cells / well.

    The method: 5
    The assay is based on the diverse ability of the chloride (Cl-) and iodide (I-) anions to extinguish the fluorescence of the YFP protein. The cells were seeded in the presence of an extracellular solution with 130 mM NaCl, a condition in which the fluorescence of the YFP is very bright. Under these conditions, the cells were incubated with the various compounds at various concentrations or with dimethyl sulfoxide (DMSO) as a control. The initial fluorescence is recorded with a fluorescence reader and after a few seconds a solution of NaI is added. If the compound is active, the I- is internalized and, competing with the Cl- for the YFP, binds to it and extinguishes the fluorescence (Figure 1). The adjustment of the curve to an exponential equation and its successive derivative allows to calculate the maximum rate of extinction of the fluorescence (VmEF) for each concentration and this parameter is a direct indication of the activity of the compound.

    Results:
    The VmEF for each concentration allows to reconstruct a dose-response curve (Figure 2) and obtain the following EC50 values: Ia> 40 µM; Ib = 21.83 ± 2.74; Ic = 17.05 ± 1.30; Id 20 = 24.88 ± 3.28. These tests show that the concentrations that allow obtaining 50% activity are lower than the concentrations to reduce the viability of 50%, especially for compounds Ib and Ic.

    It was subsequently established if the transport activity of these compounds varied with pH. For this, buffered NaI solutions were used at pH 7.3, 6.9, 6.6 and 6.2 (Figure 3). The results indicate that the compounds Ib, Ic and Id have an activity that clearly depends on the pH. For compound Ia, it is noted that the activity is very low and is not significantly diverse at the various pH values. It is interesting to note that although at pH 7.3 the activity of these compounds is much lower than that of the CFTR protein, included in the figure as a comparison, at higher acidic values the higher activity of the compounds is very close to the of the CFTR. These results suggest that in conditions of inflammation such as those described in the airways of patients with cystic fibrosis, where the pH is more acidic than in normal conditions, the compounds have an activity 2-3 times greater than that measured at pH 7.3. 35

    Another important information was to establish the kinetics of incorporation in the membrane of this type of compounds. For that, the compound Ic (12 µM) was selected because its greater activity allows a better temporary follow-up.
    The data obtained indicate that the activity of Ic increases very rapidly at the beginning, so that after 2 minutes of incubation the transport corresponds to 50% of the final activity (Figure 5 4). Transport activity continues to increase more slowly in the next 30 minutes until it reaches a more or less constant value.

    Determination of the stability of the compounds in the cell membrane. To obtain this information, the cells were incubated for 45 minutes with compound Ic (12 10 µM) and then the compound was washed from the extracellular solution and the activity was measured every two minutes for 60 minutes (Figure 5). The results show that the transport activity decreases very slowly, so that after 1 hour more than 85% of the initial activity is still detected.
     fifteen
    权利要求:
    Claims (14)
    [1]

    1. A compound of formula (I)
    R8NNNHNR5R6R7 (I) R4NR1R2R3
    where R1, R2, R3, R6, R7 and R8 are independently selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl,
    R4 and R5 are selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl,
    and wherein R9, R10, and R11 are radicals independently selected from the group consisting of: H, OH, halogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl.

    [2]
    2. The compound according to claim 1, characterized in that it is independently selected from the group consisting of:
    (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3 chloride -methoxy-2H-pyrrolium (Ia), (Z) Chloride -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H- pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy- 2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -2H-pyrrole chloride (Ic), (Z) -3-methoxy-2 - ((5-methyl-4) chloride -pentyl-1H-pyrrole-2-yl) methylene) -5- (1-octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id).

    [3]
    3. Method of obtaining a compound of formula (I) according to claims 1 or 2, comprising the following steps:
    (a) the cycloaddition reaction of an R8N3 azide and a 5-alkynyl-2H-pyrrol-2-ylidene compound of formula (III), using a copper catalyst, to generate a triazolyl-pyrrole-carbaldehyde compound of formula (II )
    (b) The condensation of the compound of formula (II) obtained above, with an unsubstituted pyrrole in the alpha position of formula (IV), in the presence of a catalytic amount of acid and an organic solvent.
    (c) Evaporation of the organic solvent and purification by chromatographic column of the compound of formula (I).

    [4]
    4. Pharmaceutical composition comprising the compound of formula (I) of claims 1 or 2, or a pharmaceutically acceptable addition salt or solvate thereof and at least one pharmaceutically acceptable excipient.

    [5]
    5. Pharmaceutical composition according to claim 4 characterized in that the compound of formula (I) is independently selected from the group consisting of: (Z) -5- (1-Butyl-1H-1,2,3) chloride -triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ia), Chloride (Z) -5- (1 -octyl-1H-1,2,3-triazol-4-yl) -2 - ((3,5-dimethyl-1H-pyrrol-2-yl) methylene) -3-methoxy-2H-pyrrolium (Ib), (Z) -5- (1-Butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-) chloride il) methylene) -2H-pyrrolium (Ic), (Z) -3-methoxy-2 - ((5-methyl-4-pentyl-1H-pyrrole-2-yl) methylene) -5- (1-) octyl-1H-1,2,3-triazol-4-yl) -2H-pyrrolium (Id).

    [6]
    6. Use of a compound of formula (I), of claims 1 or 2, to prepare a medicament.

    [7]
    7. Use of a compound of formula (I), of claims 1 or 2, to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    [8]
    8. Use of a compound of formula (I), of claims 1 or 2, to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.

    [9]
    9. Use of the pharmaceutical composition of claims 4 or 5, to prepare a medicament.

    [10]
    10. Use of the pharmaceutical composition of claims 4 or 5, to prepare a medicament for the treatment and / or prophylaxis of diseases derived from the anomalous transport of anions at the cellular level.

    [11]
    11. Use of the pharmaceutical composition of claims 4 or 5, to prepare a medicament for the treatment and / or prophylaxis of cystic fibrosis.

    [12]
    12. Intermediate of the cycloaddition reaction (a) according to claim 3, characterized in that it is a triazole-pyrrole-carbaldehyde of formula (II)
    HNR5R4NNNOR8R7R6 (II)
    where R4 and R5 are selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl,
    R6, R7 and R8 are independently selected from the group consisting of: H, OH, halogen, O-R9, NR10R11, COOR9, CONR10R11, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl,
    and wherein R9, R10 and R11 are radicals independently selected from the group consisting of: H, OH, halogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, arylalkyl, heterocycle or heteroaryl.

    [13]
    13. Intermediate compound according to claim 12, characterized in that it is 5- (1-butyl-1H-1,2,3-triazol-4-yl) -3-methoxy-1H-pyrrol-2-carbaldehyde of formula (IIa) :
    HNOHNNNOC4H9IIa

    [14]
    14. Intermediate compound according to claim 12, characterized in that it is 5- (1-octyl-1H-1,2,3-triazol-4-yl) -3-methoxy-1H-pyrrol-2-carbaldehyde of formula (IIb) .
    HNOHNNNOC8H17IIb
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    同族专利:
    公开号 | 公开日
    WO2017093593A1|2017-06-08|
    ES2615077B1|2018-03-14|
    引用文献:
    公开号 | 申请日 | 公开日 | 申请人 | 专利标题
    WO1995017381A1|1993-12-23|1995-06-29|Pharmacia S.P.A.|Novel 2,2'-bi-1h-pyrrole derivatives with immunosuppressant activity|
    WO1999040069A1|1998-02-09|1999-08-12|Pharmacia & Upjohn S.P.A.|Benzyloxy prodigiosine compounds|WO2020192904A1|2019-03-27|2020-10-01|Universidad De Burgos|2--pyrrole derivatives having ionophoric activity|
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