Azithromycin derivatives with epithelial barrier enhancement properties

Abstract

The invention provides novel compounds that are derivatives of Azithromycin and that have been found by the current inventors to have low antimicrobial activity but significant epithelial barrier enhancement properties. The invention further provides use of the compounds as a medicament, in particular in the treatment or prophylaxis of a disease or condition that is caused by a defect in epithelial cells or tissue, or a disease or condition that benefits from enhancement or restoration of epithelial barrier function, for example diseases of the respiratory tract.

Claims

1. A compound according to compound according to formula (I) ##STR00044## wherein R.sup.1 is selected from the group consisting of carbamoyloxy, NC.sub.1-6-alkylcarbamoyloxy, N(C.sub.6-14-aryl-C.sub.1-6-alkyl)carbamoyloxy, N,N-di-C.sub.1-6-alkylcarbamoyloxy, N,N-di-alkylcarbamoyloxy with the two alkyl substituents together forming a 5- to 8-membered heterocycle together with the nitrogen atom of the carbamate moiety, and C.sub.1-6-alkylcarboxy; and R.sup.2 is according to formula (III) ##STR00045## wherein R.sup.7 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.6-14-aryl-C.sub.1-6-alkyl, C.sub.3-6-alkylcarbonyl, C.sub.6-14-arylcarbonyl, C.sub.1-6-alkyl-C.sub.6-14-arylcarbonyl, C.sub.6-14-arylsulfonyl, C.sub.1-6-alkyl-C.sub.6-14-arylsulfonyl, C.sub.6-14-aryl-C.sub.1-6-alkylcarbonyl, C.sub.6-14-aryl-OC.sub.1-6-alkylcarbonyl, C.sub.6-14-aryl-C.sub.1-6-alkyl-OC.sub.1-6-alkyl-carbonyl, HOOC(CH.sub.2).sub.m-(CO)- with m being from 0 to 6, a substituent of formula (V.1) ##STR00046## with Ar being C.sub.6-14-aryl and n, p and q being independently from 0 to 6, C.sub.6-14-aryl-C.sub.1-6-alkyl-OCONHC.sub.1-6-alkyl-CO, and C.sub.6-14-arylsulfonyl, and C.sub.1-6-alkyl-C.sub.6-14-arylsulfonyl, whereby alkyl and/or aryl in R.sup.7 is optionally substituted by 1 to 6 halogen and/or CN; and R.sup.8 is selected from the group consisting of H; C.sub.6-14-arylcarbonyl optionally substituted with 1 to 5 groups selected from halogen atoms, C.sub.1-3-alkyl sulfonyl groups, C.sub.1-3-alkyl groups and/or C.sub.1-3-alkoxy groups; C.sub.3-6-alkylcarbonyl; HOOC(CH.sub.2).sub.m-(CO)- with m being from 0 to 6, a substituent of formula (V.1) as depicted hereinbefore with Ar being C.sub.6-14-aryl and n, p and q being independently from 0 to 6; and heteroarylcarbonyl having a 5- to 10-membered ring containing 1, 2 or 3 heteroatoms selected from the group consisting of O, N and S, the ring of the heteroarylcarbonyl optionally being substituted by 1 to 3 substituents selected from the group consisting of C.sub.1-4-alkyl, C.sub.1-4-alkoxy, C.sub.2-4-alkenyl, C.sub.2-4-alkenoxy, halogen and CN, whereby alkyl and/or aryl in R.sup.8 is optionally substituted by 1 to 6 halogen and/or CN; R.sup.3 is H; and R.sup.4 and R.sup.5 are independently selected from H and C.sub.1-6-alkylcarbonyl, whereby alkyl is optionally substituted by 1 to 6 halogen and/or CN; and or a pharmaceutically acceptable ester, amide, carbamate, solvate or salt thereof, including a salt of such an ester, amide or carbamate, and a solvate of such an ester, amide, carbamate or salt.

2. The compound according to claim 1, wherein R.sup.1 is selected from the group consisting of C.sub.1-3-alkylcarboxy, and a moiety according to formula (IV) ##STR00047## wherein R.sup.9 and R.sup.10 are independently selected from the group consisting of H, C.sub.1-3-alkyl and C.sub.6-10-aryl-C.sub.1-3-alkyl or are together alkyl forming a 5- or 6-membered aliphatic heterocycle together with the nitrogen atom they are both bonded to, whereby said heterocycle optionally contains one or two further heteroatom(s) selected from the group consisting of N, O and S, or said heterocycle is selected from the group consisting of piperidine, piperazine and morpholin, whereby alkyl, aryl and/or the heterocycle in R.sup.1 is optionally substituted by 1 to 6 halogen and/or CN.

3. The compound according to claim 1, wherein R.sup.7 is selected from the group consisting of C.sub.1-3-alkyl, C.sub.6-10-aryl-C.sub.1-3-alkyl, linear or branched C.sub.3-4-alkylcarbonyl, C.sub.6-10-arylcarbonyl, C.sub.6-10-aryl-C.sub.1-3-alkylcarbonyl, C.sub.6-10-aryl-OC.sub.1-3-alkylcarbonyl, C.sub.6-10-aryl-C.sub.1-3-alkyl-OC.sub.1-3-alkyl-carbonyl, HOOC(CH.sub.2).sub.m-(CO)-with m being from 0 to 3, a moiety according to formula (V.2) ##STR00048## wherein Ar is C.sub.6-10-aryl and n, p and q each are independently from 0 to 3, C.sub.6-10-aryl-C.sub.1-3-alkyl-OCONHC.sub.1-3-alkyl-CO, C.sub.6-10-arylsulfonyl, and C.sub.1-3-alkyl-C.sub.6-10-arylsulfonyl; and R.sup.8 is selected from the group consisting of H, C.sub.6-10-arylcarbonyl, C.sub.6-10-arylcarbonyl substituted with 1 to 3 halogen atoms, C.sub.1-2-alkyl groups and/or C.sub.1-2-alkoxy groups, linear or branched, C.sub.3-4-alkylcarbonyl, HOOC(CH.sub.2).sub.m-(CO)- with m being from 0 to 3, a moiety according to formula (V.2) as depicted hereinbefore wherein Ar is C.sub.6-10-aryl and n, p and q each are independently from 0 to 3, C.sub.1-3-alkylsulfonyl-bi-C.sub.6-10-aryl-carbonyl, and heteroarylcarbonyl having a 5-, 6- or 10-membered ring containing 1, 2 or 3 heteroatoms, with the heteroatom(s) in each case being selected from the group consisting of N, O and S, the ring of the heteroarylcarbonyl optionally being substituted by 1 or 2 substituents selected from the group consisting of C.sub.1-2-alkyl, C.sub.1-2-alkoxy, C.sub.2-3-alkenyl, C.sub.2-3-alkenoxy, halogen and CN; whereby alkyl, aryl and/or the heterocycle in R.sup.8 is optionally substituted by 1 to 6 halogen and/or CN.

4. The compound according to claim 1, wherein R.sup.1 is N(C.sub.6-14-aryl-C.sub.1-6-alkyl)carbamoyloxy, N,N-di-alkylcarbamoyloxy with the two alkyl substituents together forming a 5- to 8-membered heterocycle together with the nitrogen atom of the carbamate moiety and C.sub.1-6alkylcarboxy; R.sup.7 is selected from the group consisting of C.sub.1-3-alkyl, a moiety according to formula (V.2) wherein Ar is phenyl, n and p each are the same and 1 or 2 and q is 2 or 3, phenyl-C.sub.1-2-alkyl-OCONHC.sub.1-2-alkyl-COand phenyl-OC.sub.1-3-alkylcarbonyl, whereby alkyl, aryl and/or the heterocycle in R.sup.7 is optionally substituted by 1 to 3 halogen and/or CN; and R.sup.8 is selected from the group consisting of H, HOOC(CH.sub.2).sub.m-(CO)- with m being from 1 to 3, benzoyl, methylbenzoyl, ethylbenzoyl, methoxybenzoyl, ethoxybenzoyl, methylsulfonylphenylbenzoyl and naphthylcarbonyl, whereby alkyl, aryl and/or the heterocycle in R.sup.8 is optionally substituted by 1 to 3 halogen and/or CN.

5. The compound according to claim 1, wherein R.sup.1 is N(C.sub.6-14-aryl-C.sub.1-6-alkyl)carbamoyloxy, N,N-di-alkylcarbamoyloxy with the two alkyl substituents together forming a 5- to 8-membered heterocycle together with the nitrogen atom of the carbamate moiety and C.sub.1-6alkylcarboxy; R.sup.2 is according to formula (III) ##STR00049## wherein R.sup.7 is C.sub.1-6-alkyl; and R.sup.8 is selected from the group consisting of C.sub.6-14-arylcarbonyl optionally substituted with 1 to 5 groups selected from C.sub.1-3-alkyl sulfonyl groups and/or C.sub.1-3-alkyl groups; HOOC(CH.sub.2).sub.m-(CO)- with m being from 0 to 6, a substituent of formula (V.1) ##STR00050## with Ar being C.sub.6-14-aryl and n, p and q being independently from 0 to 6; R.sup.3 is H; and R.sup.4 and R.sup.5 are both H; or a pharmaceutically acceptable ester, amide, carbamate, solvate or salt thereof, including a salt of such an ester, amide or carbamate, and a solvate of such an ester, amide, carbamate or salt.

6. The compound according to claim 1, which is selected from the group consisting of (3aR,4R,7R,8S,9S, 10R, 11S, 13R, 16R, 16aR)-10-{[(2S,3R,4S,6R)-3-(benzoyloxy)-4-(dimethylamino)-6-methyloxan-2-yl]oxy}-4-ethyl-11-hydroxy-3a, 7,9,11, 13, 15, 16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-8-yl morpholine-4-carboxylate (Example 3); (2S,3R,4S,6R)-2-{[(3aR,4R,7R,8S,9S, 10R, 11S, 13R, 16R, 16aR)-8-[(benzylcarbamoyl)oxy]-4-ethyl-11-hydroxy-3a,7,9,11,13,15,16-heptamethyl-2,6-dioxo-tetradecahydro-2H-[1,3]dioxolo[4,5-c]1-oxa-6-azacyclopentadecan-10-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-yl benzoate (Example 4); (2R,3S,4R,5R,8R, 10S, 11R, 12S, 13S, 14R)-11-{[(2S,3R,4S,6R)-4-[benzyl(methyl)amino]-3-hydroxy-6-methyloxan-2-yl]oxy}-2-ethyl-3,4,10-trihydroxy-13-{[(4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8, 10, 12, 14-heptamethyl-1-oxa-6-azacyclopentadecan-15-one (Example 5); N-[(2S,3R,4S,6R)-2-{[(2R,3S,4R,5R,8R,10S,11R,12S, 13S, 14R)-2-ethyl-3,4, 10-trihydroxy-13-{[(4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12, 14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-11-yl]oxy}-3-hydroxy-6-methyloxan-4-yl]-N-methylbenzamide (Example 6); N-[(2S,3R,4S,6R)-2-{[(2R,3S,4R,5R,8R, 10S, 11R, 12S, 13S, 14R)-2-ethyl-3,4,10-trihydroxy-13-{[(4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl] oxy}-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-11-yl]oxy}-3-hydroxy-6-methyloxan-4-yl]-N-methylnaphthalene-2-sulfonamide (Example 8); N-[(2S,3R,4S,6R)-2-{[(2R,3S,4R,5R,8R, 10S, 11R, 12S, 13S, 14R)-2-ethyl-3,4,10-trihydroxy-13-{[(4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyloxan-2-yl]oxy}-3,5,6,8,10,12, 14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-11-yl]oxy}-3-hydroxy-6-methyloxan-4-yl]-N,4-dimethylbenzene-1-sulfonamide (Example 10); (2S,3R,4S,6R)-2-{[(2R,3R,4R,5R,8R, 10S, 11R, 12S, 13S, 14R)-4, 13-bis(acetyloxy)-2-ethyl-3,10-dihydroxy-3,5,6,8,10,12,14-heptamethyl-15-oxo-1-oxa-6-azacyclopentadecan-11-yl]oxy}-4-(dimethylamino)-6-methyloxan-3-yl benzoate (Example 11).

7. A pharmaceutical composition comprising at least one compound of Formula (Ia) and at least one pharmaceutically acceptable excipient: ##STR00051## wherein R.sup.1 is selected from the group consisting of carbamoyloxy, NC.sub.1-6-alkylcarbamoyloxy, N(C.sub.6-14-aryl-C.sub.1-6-alkyl)carbamoyloxy, N,N-di-C.sub.1-6-alkylcarbamoyloxy, N,N-di-alkylcarbamoyloxy with the two alkyl substituents together forming a 5- to 8-membered heterocycle together with the nitrogen atom of the carbamate moiety, and C.sub.1-6-alkylcarboxy; and R.sup.2 is according to formula (III) ##STR00052## wherein R.sup.7 is selected from the group consisting of C.sub.1-6-alkyl, and R.sup.8 is selected from the group consisting of H; C.sub.6-14-arylcarbonyl optionally substituted with 1 to 5 groups selected from halogen atoms, C.sub.1-3-alkyl sulfonyl groups, C.sub.1-3-alkyl groups and/or C.sub.1-3-alkoxy groups; C.sub.3-6-alkylcarbonyl; HOOC(CH.sub.2).sub.m-(CO)- with m being from 0 to 6, a substituent of formula (V.1) as depicted hereinbefore with Ar being C.sub.6-14-aryl and n, p and q being independently from 0 to 6; and heteroarylcarbonyl having a 5- to 10-membered ring containing 1, 2 or 3 heteroatoms selected from the group consisting of O, N and S, the ring of the heteroarylcarbonyl optionally being substituted by 1 to 3 substituents selected from the group consisting of C.sub.1-4-alkyl, C.sub.1-4-alkoxy, C.sub.2-4-alkenyl, C.sub.2-4-alkenoxy, halogen and CN, whereby alkyl and/or aryl in R.sup.8 is optionally substituted by 1 to 6 halogen and/or CN; R.sup.3 is H; and R.sup.4 and R.sup.5 are independently selected from H and C.sub.1-6-alkylcarbonyl, whereby alkyl is optionally substituted by 1 to 6 halogen and/or CN; or a pharmaceutically acceptable ester, amide, carbamate, solvate or salt thereof, including a salt of such an ester, amide or carbamate, and a solvate of such an ester, amide, carbamate or salt.

8. A method for the treatment of a disease or condition that benefits from enhancement or restoration of epithelial barrier function, said method comprising administering to a mammal a therapeutically effective amount of the compound of claim 1.

9. The method according to claim 8, wherein the disease or condition is an airways disease and the epithelial tissue is in the respiratory tract epithelial tissue.

10. The method according to claim 8 wherein the condition is asthma, chronic obstructive pulmonary disease (COPD), Cystic Fibrosis (CF), non-CF Bronchiectasis, chronic rhinosinusitis, diffuse panbronchiolitis (DPB), chronic bronchitis, Bronchiolitis Obliterans Organizing Pneumonia (BOOP) primary or secondary to chemotherapy or post-transplantation status, infantile respiratory distress syndrome (IRDS) and its long term complication, bronchopulmonary dysplasia, neuromuscular respiratory depression and/or failure, pneumonia and conditions caused by and associated with Respiratory Syncytial Virus (RSV) and related viruses.

11. The method according to claim 8 wherein the treatment of a disease or disorder is systemic inflammatory distress syndrome (SIRS), adult respiratory distress syndrome (ARDS), inflammatory bowel disease, ulcerative colitis or Crohn's disease.

12. A method for the treatment or prophylaxis of a disease or condition that benefits from enhancement or restoration of epithelial barrier function, which comprises administering to the mammal a therapeutically effective amount of a compound or composition as defined in claim 1.

Description

Example D: Barrier Integrity Enhancement in the Lung

(1) The effect of compounds of the invention on maintenance of barrier integrity in the lungs of mice under sulphur dioxide insult was investigated.

(2) Method:

(3) 1. 15 female mice were randomised into 3 groups of 5 mice. The three groups were: a) Control group b) Azithromycin treatment group c) Investigational compound treatment group

(4) 2. The mice were pre-treated for three days with: a) Placebo (Phosphate buffered salinePBS) given at a dose of 10 mg/kg in an intravenous injection of 100 ?l three times per week. b) Azithromycin: given at a dose of 10 mg/kg in an intravenous injection of 100 ?l three times per week. c) The compound of Example 2: given at a dose of 10 mg/kg in an intravenous injection of 100 ?l three times per week.

(5) 3. One mouse in each group was set to one side as a control. Four mice were then exposed to sulphur dioxide gas at a concentration of 200 ppm in air for a period of 60 minutes. The mice were in a cage that was inserted into a 45 L treatment box with a removable lid, a gas feed line and a gas exhaust vent. Gas was fed into the box at a rate that resulted in roughly 7 volume changes per hour. The gas was premixed from bottled air and bottled 200 ppm SO.sub.2 in a first box and then fed into the treatment box. The level of SO.sub.2 being fed into the treatment box can be varied by adjusting the relative flow of air and SO.sub.2 from the two source cylinders. As the sulphur dioxide concentration in this experiment was 200 ppm, it was used neat, with no additional air being mixed with it. The mice had access to food and water for the duration of the treatment.

(6) 4. 24 hours after the end of the treatment, 1 mg of human serum albumin (HSA) in 100p saline was injected into the tail vein of each mouse, and fluorescent dextrans (FDs) were injected into the airways. The mice were then kept sedated for 90 minutes.

(7) 5. After 90 minutes, the mice were sacrificed via exsanguination. Plasma and bronchoalveolar lavage (BAL) were collected and the lungs were processed for paraffin embedding.

(8) 6. The level of HSA in the BAL was quantified by ELISA. The ELISA test used was the one commercially available from abcam (Human Serum Albumin kit ab179887). In line with the kit manufacturer's instructions, absorbance at 600 nm was measured. Fluorescent dextrans are quantified using a fluorescent plate reader.

(9) The level of HSA that leaks into the lungs, as found in the BAL, is a measure of the integrity of the lung epithelial barrier.

(10) Results:

(11) In the placebo group, there were experimental difficulties with two of the four mice; in the Azithromycin group, there were experimental difficulties with one of the four mice; and in the investigational compound group, there were experimental difficulties with two of the four mice. Therefore, results were obtained from two mice in each of the placebo and investigational compound groups, and from three mice in the Azithromycin group. The HSA data read outs were as follows:

(12) TABLE-US-00004 HSA level- HSA level- absorbance at absorbance 600 nm relative Group at 600 nm to control PBS Control 0.584 ? 0.175 1.000 ? 0.300 Azithromycin 0.346 ? 0.031 0.593 ? 0.053 Example 2 0.373 ? 0.037 0.640 ? 0.058

(13) The data are shown in the bar chart in FIG. 1.

(14) It is seen that for both azithromycin and the compound of Example 2, the leakage of HSA into the lung following SO.sub.2 insult, was reduced compared with the control. This indicates that azithromycin and the compound of Example 2 enhance the resilience of the barrier function of epithelial cells in the lung.