NEW CLASS OF ANTIBIOTICS HAVING LOW MIC-VALUES TOWARDS DIFFERENT STRAINS OF BACTERIA
20220313656 · 2022-10-06
Inventors
- Jørn B. Christensen (Virum, DK)
- Rikke Heidemann Olsen (Hvidovre, DK)
- Søren Wedel Svenningsen (København K, DK)
Cpc classification
C07D335/20
CHEMISTRY; METALLURGY
C07D279/28
CHEMISTRY; METALLURGY
C07D417/06
CHEMISTRY; METALLURGY
C07D279/26
CHEMISTRY; METALLURGY
Y02A50/30
GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
International classification
A61K31/5415
HUMAN NECESSITIES
C07D279/28
CHEMISTRY; METALLURGY
Abstract
The present invention relates to a composition comprising a compound of formula (I) wherein X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl; Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl; each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; d is selected from 0, 1, 2, and 3; each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S-C3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; e is selected from 0, 1, 2, 3, and 4; R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl; R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3); each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where; Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl; Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl; Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl; where A is selected from any pharmaceutical relevant/acceptable anion/counterion; wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl; wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl. The invention also relates to anti-microbial composition for use as a medicament and for use in treating a microbial infection in a human subject.
##STR00001##
Claims
1. A composition comprising a compound of formula I: ##STR00047## wherein: X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 and C.sub.0-2-alkyl; Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, and COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl; each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; d is selected from 0, 1, 2, or 3; each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; e is selected from 0, 1, 2, 3, or 4; R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl; R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3); each W is individually selected from linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where; Y.sup.1 is selected from C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl; Y.sup.2 is selected from C.sub.1-12-alkyl; Y.sup.3 is selected from linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl; where A is selected from any pharmaceutical acceptable anion or counterion; wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl; and wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl.
2-32. (canceled)
33. The composition according to claim 1 wherein the compound of formula I is a phenothiazine derivative selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof or wherein the compound of formula I is a chlorprothixene derivative.
34. The composition according to claim 1 wherein Y.sup.3 is a linear or branched C.sub.5-25-alkyl.
35. The composition according to claim 1 wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl.
36. The composition according to claim 1 wherein Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl.
37. The composition according to claim 1 wherein Y.sup.1 and Y.sup.2 are both C.sub.1-alkyl.
38. The composition according to claim 1 wherein Y.sup.1 together with a carbon being part of the W and the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl.
39. The composition according to claim 1 wherein Z is selected from hydrogen, Cl or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl.
40. A method of inhibiting a microbial infection in a subject comprising: administering the composition of claim 1 to a subject that has a microbial infection.
41. The method according to claim 40, wherein the compound of formula I is a phenothiazine derivative selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof or wherein the compound of formula I is a chlorprothixene derivative.
42. The method according to claim 40, wherein Y.sup.3 is a linear or branched C.sub.5-25-alkyl.
43. The method according to claim 40, wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl.
44. The method according to claim 40, wherein the microbial infection comprises a resistant strain of bacteria selected from Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria, Escherichia or Salmonella.
45. The method according to claim 40, wherein composition provides a minimum inhibitory concentration (MIC) below 16 μg/mL.
46. The composition of claim 34 wherein Y.sup.3 is a linear or branched C.sub.8-15-alkyl.
47. The composition of claim 34 wherein Y.sup.3 is selected from the group consisting of a linear or branched C.sub.8-alkyl, a linear or branched C.sub.10-alkyl, a linear or branched C.sub.12-alkyl, a linear or branched C.sub.14-alkyl, and a linear or branched C.sub.15-alkyl.
48. The composition of claim 35 wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl and pentyl.
49. The method of claim 42 wherein Y.sup.3 is a linear or branched C.sub.8-15-alkyl.
50. The method of claim 42 wherein Y.sup.3 is selected from the group consisting of a linear or branched C.sub.8-alkyl, a linear or branched C.sub.10-alkyl, a linear or branched C.sub.12-alkyl, a linear or branched C.sub.14-alkyl, and a linear or branched C.sub.15-alkyl.
51. The method of claim 43 wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl and pentyl.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0077] The invention is further illustrated by the drawings, wherein:
[0078]
[0079]
[0080]
[0081]
[0082]
[0083]
[0084]
DETAILED DESCRIPTION OF THE INVENTION
[0085] The inventors have conducted intensive studies on a large range of compounds in an attempt to find new antibacterial drugs and has found unprecedented findings that compounds of the below given formula shows promising results in regards to having properties relevant for antimicrobial activity.
[0086] A first aspect of the invention defines a composition comprising a compound of formula I
##STR00005##
[0087] wherein
[0088] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl;
[0089] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl;
[0090] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0091] d is selected from 0, 1, 2, and 3;
[0092] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0093] e is selected from 0, 1, 2, 3, and 4;
[0094] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl;
[0095] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3);
[0096] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where;
[0097] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the Wand the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl;
[0098] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl;
[0099] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl;
[0100] where A is selected from any pharmaceutical relevant/acceptable anion/counterion; wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl;
[0101] wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl.
[0102] In one embodiment, X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.2 or C.sub.0-2-alkyl.
[0103] It is surprising that the inventors have found out that this specific group of compounds all have antimicrobial activity towards different strain of bacteria including resistant strains.
[0104] The inventors managed to synthesize several modifications of thioridazine, and in addition, the same modifications were done to other phenothiazine-derivatives and to chlorprothixene.
[0105] The results shows that by modulating the chemical structure of thioridazine and of the other phenothiazine-derivatives and of chlorprothixene, the compounds were less permeable to the blood-brain barrier. Further the compounds showed between 2-128 fold higher antibiotic activities (defined by the lowest concentration of the compound that allow inhibition of bacterial growth, also known as the minimal inhibitory concentration (MIC) value) than the original hydrochloride-state of the same compound.
[0106] The compounds are quaternized on the side-chain nitrogen with different alkyl groups. The length of the chain has surprisingly shown to be important.
[0107] Where nothing else is mentioned or written in the definitions of the formulas herein, it is implicit that there is a hydrogen atom attached. For example; if d is selected from 0, 1 or 2, so that a R.sup.2 group is positioned at 0, 1 or 2 places—there will be a hydrogen atom attached to the remaining places. Also in the formula definition for R.sup.5: N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3), it is implicit that there is a hydrogen atom attached to “fill up”, so that there won't be any radicals.
[0108] In one embodiment the compound of formula I is a phenothiazine derivative. The phenothiazine derivative can be selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof.
[0109] In one embodiment the compound of formula I is a chlorprothixene derivative.
[0110] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0111] In one or more embodiment, there is at least three carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0112] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a branched C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH(CH.sub.3))—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0113] In one or more embodiment, there is at least three carbon atoms between the carbon atom in the ring structure and the nitrogen atom. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then C═CH—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0114] In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.2-25 alkenyl. In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25 alkynyl. In one embodiment Y.sup.3 is a linear or branched C.sub.3-25 aliphatic group.
[0115] In one embodiment, Y.sup.3 is a linear or branched C.sub.5-25-alkyl. In one embodiment, Y.sup.3 is a linear or branched alkyl with a side chain higher than C.sub.5.
[0116] In one embodiment Y.sup.3 is a linear or branched C.sub.2-6-alkyl. Y.sup.3 can be selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl.
[0117] In one embodiment Y.sup.3 is a linear or branched C.sub.8-15-alkyl. Y.sup.3 can be selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl.
[0118] In one embodiment Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. In one embodiment Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl.
[0119] In one embodiment Y.sup.1 together with a carbon being part of the Wand the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl. Y.sup.2 can then in one embodiment be selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl.
[0120] In one embodiment X is S. in one embodiment Z can be selected from the group consisting of hydrogen, Cl or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl.
[0121] The pharmaceutical relevant/acceptable anion/counterion can be any suitable known relevant/acceptable anion/counterion and in one embodiment it can be selected from the group consisting of I, Br or Cl, MS or TS.
[0122] In one embodiment the compound of formula I is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethylpentan-1-aminium bromide.
[0123] In one embodiment the compound of formula I is a promethazine derivative such as N-isopropyl-N,N-dimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium bromide.
[0124] In one embodiment the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0125] In one embodiment the compound is a promethazine derivative such as N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyldodecan-1-aminium bromide.
[0126] In one embodiment the compound of formula I is a thioridazine derivative such as 1-ethyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment the compound is a thioridazine derivative such as 1-isopropyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment the compound is a thioridazine derivative such as 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment the compound is a thioridazine derivative such as 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide. In one embodiment the compound is a thioridazine derivative such as 1-dodecyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium.
[0127] In one embodiment the compound of formula I is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethylpentan-1-aminium bromide. In one embodiment the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0128] In one embodiment the compound is N-(3-(10H-phenothiazin-10-yl)propyl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0129] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyltetradecan-1-aminium bromide.
[0130] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0131] A second aspect of the invention defines an anti-microbial composition comprising a compound of formula I
##STR00006##
[0132] wherein
[0133] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl;
[0134] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl;
[0135] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0136] d is selected from 0, 1, 2, and 3;
[0137] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0138] e is selected from 0, 1, 2, 3, and 4;
[0139] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl;
[0140] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3);
[0141] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where;
[0142] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl;
[0143] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl;
[0144] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl; linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl;
[0145] where A is selected from any pharmaceutical relevant/acceptable anion/counterion; for use as a medicament.
[0146] In one embodiment X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.2 or C.sub.0-2-alkyl.
[0147] In one embodiment of the second aspect, the compound of formula I in the anti-microbial composition is a phenothiazine derivative. The phenothiazine derivative can be selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof. In one embodiment the compound of formula I is a chlorprothixene derivative.
[0148] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0149] In one or more embodiment, there is at least three carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0150] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a branched C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH(CH.sub.3))—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0151] In one or more embodiment, there is at least three carbon atoms between the carbon atom in the ring structure and the nitrogen atom. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then C═CH—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0152] In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.2-25 alkenyl. In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25 alkynyl. In one embodiment Y.sup.3 is a linear or branched C.sub.3-25 aliphatic group.
[0153] In one embodiment, Y.sup.3 is a linear or branched C.sub.5-25-alkyl. In one embodiment, Y.sup.3 is a linear or branched alkyl with a side chain higher than C.sub.5.
[0154] In one embodiment Y.sup.3 is a linear or branched C.sub.2-6-alkyl. Y.sup.3 can be selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. In one embodiment Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. In one embodiment Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.2-6-alkyl. Y.sup.3 can be selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. Y.sup.1 and Y.sup.2 can be individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl in one embodiment.
[0155] In one embodiment Y.sup.3 is a linear or branched C.sub.8-15-alkyl. Y.sup.3 can be selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl. In one embodiment Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. In one embodiment Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.8-12-alkyl. Y.sup.3 can be selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.11-alkyl or linear or branched C.sub.12-alkyl. Y.sup.1 and Y.sup.2 can be individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl in one embodiment.
[0156] In one embodiment the anti-microbial composition comprise that Y.sup.1 together with a carbon being part of the Wand the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl and Y.sup.2 can be selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl.
[0157] In one embodiment of the anti-microbial composition X is S. In one embodiment Z is selected from the group consisting of hydrogen, C.sub.1 or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl.
[0158] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against resistant strains of bacteria (resistant towards conventional antimicrobial). These resistant strains of bacteria can be selected from the genus of Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria, Escherichia or Salmonella. The resistant strains of bacteria can also be selected from the genus of Clostridium, Cutibacterium or Campylobacter.
[0159] Hence, the anti-microbial composition for use as a medicament according to the second aspect can be used to treat an infection, which is caused by the selected bacteria.
[0160] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against Gram-positive bacteria. These Gram-positive bacteria can be selected from the genus of Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria or Cutibacterium, such as selected from Staphylococcus aureus, Bacillus cereus, Enterococcus faecium, Enterococcus faecalis, Staphylococcus pseudintermedius, Staphylococcus epidemidis, Streptococcus equi, Cutibacterium acnes, Clostridium difficile, Listeria monocytogenes strains.
[0161] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against certain Gram-negative bacteria. These Gram-negative bacteria can be selected from the genus of Campylobacter, Escherichia or Salmonella such as selected from Campylobacter jejuni, Escherichia coli or Salmonella Enteritidis strains.
[0162] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against Staphylococcus epidermidis.
[0163] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against Campylobacter jejuni.
[0164] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against Cutibacterium acnes.
[0165] The anti-microbial composition for use as a medicament according to the second aspect can have antibacterial properties against multi-resistant strains of bacteria. The multi-resistant strains of bacteria can be selected from Staphylococcus aureus Bacillus cereus, Staphylococcus epidermidis, Clostridium difficile, Enterococcus faecalis, or Enterococcus faecium.
[0166] The anti-microbial composition for use as a medicament according to the present disclosure can have a minimum inhibitory concentration (MIC) below 16 μg/mL, such as below 8 μg/mL, such as below 4 μg/mL or such as below 2 μg/mL.
[0167] In one embodiment of the second aspect the compound of formula I is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment of the second aspect the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethylpentan-1-aminium bromide. In one embodiment of the second aspect the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0168] In one embodiment of the second aspect the compound of formula I is a promethazine derivative such as N-isopropyl-N,N-dimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium bromide. In one embodiment of the second aspect the compound is a promethazine derivative such as N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyldodecan-1-aminium bromide.
[0169] In one embodiment of the second aspect the compound of formula I is a thioridazine derivative such as 1-ethyl-1-methyl-2-(2-(2-(methylthio)-1-OH-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the second aspect the compound is a thioridazine derivative such as 1-isopropyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the second aspect the compound is a thioridazine derivative such as 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the second aspect the compound is a thioridazine derivative such as 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide. In one embodiment of the second aspect the compound is a thioridazine derivative such as 1-dodecyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide.
[0170] In one embodiment of the second aspect the compound of formula I is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment of the second aspect the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethylpentan-1-aminium bromide. In one embodiment of the second aspect the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0171] In one embodiment the compound is N-(3-(10H-phenothiazin-10-yl)propyl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0172] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyltetradecan-1-aminium bromide.
[0173] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0174] A third aspect of the invention defines an anti-microbial composition comprising a compound of formula I
##STR00007##
[0175] wherein
[0176] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl;
[0177] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl;
[0178] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0179] d is selected from 0, 1, 2, and 3;
[0180] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy;
[0181] e is selected from 0, 1, 2, 3, and 4;
[0182] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl;
[0183] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3);
[0184] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where;
[0185] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl;
[0186] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl;
[0187] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, Hear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl;
[0188] where A is selected from any pharmaceutical relevant/acceptable anion/counterion; for use in treating a microbial infection in a human subject.
[0189] In one embodiment X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.2 or C.sub.0-2-alkyl.
[0190] In one embodiment of the third aspect the compound of formula I in the anti-microbial composition is a phenothiazine derivative. The phenothiazine derivative can be selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof. In one embodiment the compound of formula I is a chlorprothixene derivative.
[0191] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0192] In one or more embodiment, there is at least three carbon atoms between the two nitrogen atoms. In the case where the W is a C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0193] In one or more embodiment, there is at least two carbon atoms between the two nitrogen atoms. In the case where the W is a branched C.sub.2-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then N—(CH.sub.2CH(CH.sub.3))—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0194] In one or more embodiment, there is at least three carbon atoms between the carbon atom in the ring structure and the nitrogen atom. In the case where the W is a C.sub.1-alkyl group attached to both the CH group and the N(Y.sup.1)(Y.sup.2)(Y.sup.3) group —R.sup.5 is then C═CH—(CH.sub.2CH.sub.2)—N(Y.sup.1)(Y.sup.2)(Y.sup.3).
[0195] In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.2-25 alkenyl. In one embodiment, Y.sup.3 is a linear or branched C.sub.2-25 alkynyl. In one embodiment Y.sup.3 is a linear or branched C.sub.3-25 aliphatic group.
[0196] In one embodiment, Y.sup.3 is a linear or branched C.sub.5-25-alkyl. In one embodiment, Y.sup.3 is a linear or branched alkyl with a side chain higher than C.sub.5.
[0197] In one embodiment Y.sup.3 is a linear or branched C.sub.2-6-alkyl. Y.sup.3 can be selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. In one embodiment Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. In one embodiment Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.2-6-alkyl. Y.sup.3 can be selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. Y.sup.1 and Y.sup.2 can be individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl in one embodiment.
[0198] In one embodiment Y.sup.3 is a linear or branched C.sub.8-15-alkyl. Y.sup.3 can be selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl. In one embodiment Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. In one embodiment Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl. In one embodiment Y.sup.3 is a linear or branched C.sub.8-12-alkyl. Y.sup.3 can be selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.11-alkyl or linear or branched C.sub.12-alkyl. Y.sup.1 and Y.sup.2 can be individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. Y.sup.1 and Y.sup.2 can both be C.sub.1-alkyl in one embodiment.
[0199] In one embodiment the anti-microbial composition comprise that Y.sup.1 together with a carbon being part of the W and the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl and Y.sup.2 can be selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl.
[0200] In one embodiment of the anti-microbial composition X is S. In one embodiment Z is selected from the group consisting of hydrogen, C.sub.1 or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl.
[0201] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against resistant (towards conventional antimicrobials) and non-resistant strains of bacteria. The resistant strains of bacteria can be selected from the genus of Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria, Escherichia or Salmonella. The resistant strains of bacteria can also be selected from the genus of Clostridium, Cutibacterium or Campylobacter.
[0202] Hence, the anti-microbial composition for use as a medicament according to the third aspect can be used to treat an infection, which is caused by the selected bacteria.
[0203] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against Gram-positive bacteria. These Gram-positive bacteria can be selected from the genus of Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria or Cutibacterium, such as selected from Staphylococcus aureus, Bacillus cereus, Enterococcus faecium, Enterococcus faecalis, Staphylococcus pseudintermedius, Staphylococcus epidemidis, Cutibacterium acnes, Streptococcus equi, Clostridium difficile, Listeria monocytogenes strains.
[0204] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against certain Gram-negative bacteria. These Gram-negative bacteria can be selected from the genus of Campylobacter, Escherichia or Salmonella such as selected from Campylobacter jejuni, Escherichia coli or Salmonella Enteritidis strains.
[0205] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against Staphylococcus epidermidis.
[0206] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against Campylobacter jejuni.
[0207] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against Cutibacterium acnes.
[0208] The anti-microbial composition for use as a medicament according to the third aspect can have antibacterial properties against multi-resistant strains of bacteria. The multi-resistant strains of bacteria can be selected from Staphylococcus aureus, Bacillus cereus, Staphylococcus epidermidis, Clostridium difficile, Enterococcus faecalis, or Enterococcus faecium.
[0209] The anti-microbial composition for use as a medicament according to the present disclosure can have a minimum inhibitory concentration (MIC) below 16 μg/mL, such as below 8 μg/mL, such as below 4 μg/mL or such as below 2 μg/mL.
[0210] In one embodiment of the third aspect the compound of formula I is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment of the third aspect the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethylpentan-1-aminium bromide. In one embodiment of the third aspect the compound is a chlorpromazine derivative such as N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0211] In one embodiment of the third aspect the compound of formula I is a promethazine derivative such as N-isopropyl-N,N-dimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium bromide. In one embodiment of the third aspect the compound is a promethazine derivative such as N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyldodecan-1-aminium bromide.
[0212] In one embodiment of the third aspect the compound of formula I is a thioridazine derivative such as 1-ethyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the third aspect the compound is a thioridazine derivative such as 1-isopropyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the third aspect the compound is a thioridazine derivative such as 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide. In one embodiment of the third aspect the compound is a thioridazine derivative such as 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide. In one embodiment of the third aspect the compound is a thioridazine derivative such as 1-dodecyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide.
[0213] In one embodiment of the third aspect the compound of formula I is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N,3-trimethylbutan-1-aminium bromide. In one embodiment of the third aspect the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethylpentan-1-aminium bromide. In one embodiment of the third aspect the compound is a chlorprothixene derivative such as (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethyldodecan-1-aminium bromide.
[0214] In one embodiment the compound is N-(3-(10H-phenothiazin-10-yl)propyl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0215] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyltetradecan-1-aminium bromide.
[0216] In one embodiment the compound is N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate.
[0217] In one embodiment the microbial infection can be a topical or intestinal infection. In one embodiment, the topical infection can be acne.
[0218] As example 7 shows, the compound 43 was evaluated for efficacy in an MRSA skin infection model and showed to be highly effective in decreasing the bacterial load of MRSA.
EXAMPLES
[0219] All analytical values specified as ratio and in percent are by weight.
Example 1
[0220] General procedure for removal of HCl salt:
##STR00008##
[0221] The tertiary amine hydrochloride salt of Chlorpromazine, Thioridazine, Promethazine or Promazine (2.00 g; 4.91 mmol-6.25 mmol) was added to a round bottomed flask equipped with a magnetic stirring bar. Et20 (40 ml) and NaOH (2M) (40 ml) was added and the two phase system was stirred vigorously overnight. The two phases were separated and the water phase was extracted with ether (50 ml). The combined ether phase was dried over MgSO.sub.4, filtered and evaporated in vacuo giving the free tertiary amine.
[0222] Chlorpromazine: Yield: 1.55 g (87%)
[0223] Thioridazine: Yield: 1.71 g (94%)
[0224] Promethazine: Yield: 1.65 g (93%)
[0225] Promazine: Yield: 1.68 g (95%)
Example 2
[0226] General synthesis procedure for the quaternization of the tertiary amine:
##STR00009##
[0227] The free tertiary amine was added to a round bottomed flask equipped with a magnetic stirring bar. A solution of the alkyl halide in acetonitrile was added and refluxed in the dark overnight. The solution was evaporated in vacuo and dissolved in a min. of methanol. The methanol solution was dropped out in diethyl ether with stirring. The suspension was stirred until clear ether phase, and the ether was decanted off. Residues of ether were removed from the precipitate by high vacuum.
[0228] All the synthesized compounds are verified by either NMR and/or by HRMS (not shown).
[0229] Chlorpromazine hydrochloride (S1) (see table 1) is the starting material bought from commercial supplier.
[0230] 3-(2-chloro-10H-phenothiazin-10-yl)-N,N,N-trimethylpropan-1-aminium iodide (S2):
[0231] Chlorpromazine (1.00 g; 3.14 mmol), Methyl iodide (2.0 ml; 32 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.44 g (99%)
[0232] 3-(2-chloro-10H-phenothiazin-10-yl)-N-ethyl-N,N-dimethylpropan-1-aminium bromide (S3):
[0233] Chlorpromazine (1.00 g; 3.14 mmol), Ethyl bromide (2.0 ml; 27 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.25 g (93%)
[0234] 3-(2-chloro-10H-phenothiazin-10-yl)-N-isopropyl-N,N-dimethylpropan-1-aminium bromide (S4):
[0235] Chlorpromazine (0.50 g; 1.6 mmol),2-Bromopropane (4.0 ml; 43 mmol), Acetonitrile (16 ml), Diethyl ether (200 ml), Yield: 0.69 g (100%)
[0236] N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N,3-trimethylbutan-1-aminium bromide (S5):
[0237] Chlorpromazine (0.48 g; 1.15 mmol), 1-bromo-3-methylbutane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.54 g (76%)
[0238] N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethylpentan-1-aminium bromide (S6):
[0239] Chlorpromazine (0.38 g; 1.19 mmol), 1-bromopentane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.16 g (29%)
[0240] N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethyldodecan-1-aminium bromide (S7):
[0241] Chlorpromazine (0.56 g; 1.76 mmol), 1-bromododecane (3.0 ml; 13 mmol), Acetonitrile (17 ml).
[0242] After reflux, the solution was concentrated in vacuo and purified by a silica plug (Silica: 60 Å, 15-40 μm). (Eluent: Heptane, released with DCM/MeOH (1:1)). The solution was evaporated in vacuo giving the wanted compound. Yield: 0.91 g (91%)
[0243] Promethazine hydrochloride (S12) (see table 2) is the starting material bought from commercial supplier.
[0244] N,N,N-trimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium iodide (S13):
[0245] Promethazine (1.00 g; 3.52 mmol) Methyl iodide (2.0 ml; 32 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.48 g (99%)
[0246] N-ethyl-N,N-dimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium bromide (S14):
[0247] Promethazine (1.00 g; 3.52 mmol) Ethyl bromide (2.0 ml; 27 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.03 g (94%)
[0248] N-isopropyl-N,N-dimethyl-1-(10H-phenothiazin-10-yl)propan-2-aminium bromide (S15):
[0249] Promethazine (0.50 g; 1.8 mmol) 2-Bromopropane (4.0 ml; 43 mmol), Acetonitrile (16 ml), Diethyl ether (200 ml), Yield: 0.10 g (14%)
[0250] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N,3-trimethylbutan-1-aminium bromide (S16):
[0251] Promethazine (0.57 g; 2.00 mmol), 1-bromo-3-methylbutane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.16 g (68%)
[0252] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethylpentan-1-aminium bromide (S17):
[0253] Promethazine (0.64 g; 2.25 mmol), 1-bromopentane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.91 g (93%)
[0254] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyldodecan-1-aminium bromide (S18):
[0255] Promethazine (0.78 g; 2.74 mmol), 1-bromododecane (3.0 ml; 13 mmol), Acetonitrile (17 ml). After reflux, the solution was concentrated in vacuo and purified by a silica plug (Silica: 60 Å, 15-40 μm). (Eluent: Heptane, released with DCM/MeOH (1:1)). The solution was evaporated in vacuo giving the wanted compound. Yield: 1.37 g (94%)
[0256] Thioridazine hydrochloride (S23) (see table 3) is the starting material bought from commercial supplier.
[0257] 1,1-dimethyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium iodide (S24):
[0258] Thioridazine (1.00 g; 2.70 mmol) Methyl iodide (2.0 ml; 32 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.37 g (99%)
[0259] 1-ethyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S25):
[0260] Thioridazine (1.00 g; 2.70 mmol) Ethyl bromide (2.0 ml; 27 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 1.24 g (96%)
[0261] 1-isopropyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S26):
[0262] Thioridazine (0.50 g; 1.3 mmol) 2-Bromopropane (4.0 ml; 43 mmol), Acetonitrile (16 ml), Diethyl ether (200 ml), Yield: 0.09 g (13%)
[0263] 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S27):
[0264] Thioridazine (0.55 g; 1.48 mmol), 1-bromo-3-methylbutane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.16 g (27%)
[0265] 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide (S28):
[0266] Thioridazine (0.62 g; 1.67 mmol), 1-bromopentane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (200 ml), Yield: 0.44 g (51%)
[0267] 1-dodecyl-1-methyl-2-(2-(2-(methylthio)-1-OH-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S29):
[0268] Thioridazine (0.90 g; 2.43 mmol), 1-bromododecane (3.0 ml; 13 mmol), Acetonitrile (17 ml). After reflux, the solution was concentrated in vacuo and purified by a silica plug (Silica: 60 Å, 15-40 μm). (Eluent: Heptane, released with DCM/MeOH (1:1)). The solution was evaporated in vacuo giving the wanted compound. Yield: 1.50 g (99%)
[0269] Chlorprothixene (S34) (see table 4) is the starting material bought from commercial supplier.
[0270] (Z)-3-(2-chloro-9H-thioxanthen-9-ylidene)-N,N,N-trimethylpropan-1-aminium iodide (S35):
[0271] Chlorprothixene (0.15 g; 0.47 mmol), Methyl iodide (2.0 ml; 32 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.22 g (100%) (Alkyl halide: methyl iodide)
[0272] (Z)-3-(2-chloro-9H-thioxanthen-9-ylidene)-N-ethyl-N,N-dimethylpropan-1-aminium bromide (S36):
[0273] Chlorprothixene (0.15 g; 0.47 mmol), Ethyl bromide (2.0 ml; 27 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.20 g (100%) (Alkyl halide: Ethyl bromide)
[0274] (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N,3-trimethylbutan-1-aminium bromide (S37):
[0275] Chlorprothixene (0.15 g; 0.47 mmol), 1-bromo-3-methylbutane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.22 g (100%) (Alkyl halide: Pentyl bromide)
[0276] Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethylpentan-1-aminium bromide (S38):
[0277] Chlorprothixene (0.15 g; 0.47 mmol), 1-bromopentane (2.0 ml; 16 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.22 g (100%) (Alkyl halide: 1-Bromo-3-methylbutane)
[0278] (Z)—N-(3-(2-chloro-9H-thioxanthen-9-ylidene)propyl)-N,N-dimethyldodecan-1-aminium bromide (39):
[0279] Chlorprothixene (0.05 g; 0.16 mmol), 1-bromododecane (1.0 ml; 4.2 mmol), Acetonitrile (9 ml). After reflux, the solution was concentrated in vacuo and purified by a silica plug (Silica: 60 Å, 15-40 μm). (Eluent: Heptane, released with DCM/MeOH (1:1)). The solution was evaporated in vacuo giving the wanted compound. Yield: 0.09 g (100%)
[0280] N-(3-(2-chloro-10H-phenothiazin-10-yl)propyl)-N,N-dimethylprop-2-en-1-aminium bromide (S40):
[0281] Chlorpromazine (0.20 g; 0.63 mmol), allyl bromide (2.0 ml; 23 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.27 g (96%).
[0282] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethylprop-2-en-1-aminium bromide (S41):
[0283] Promethazine (0.20 g; 0.70 mmol), Allyl bromide (2.0 ml; 23 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.25 g (86%)
[0284] 1-allyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S42):
[0285] Thioridazine (0.20 g; 0.54 mmol), Allyl bromide (2.0 ml; 23 mmol), Acetonitrile (18 ml), Diethyl ether (100 ml), Yield: 0.23 g (85%).
[0286] N-(3-(10H-phenothiazin-10-yl)propyl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate (S43):
[0287] Promazine (1.0 g; 3.3 mmol), 3,7-Dimethyloct-6-en-1-yl 4-methylbenzenesulfonate (1.2 g; 3.8 mmol) (synthesized by the procedure of P. H. G. Zarbin, A. Rreckziegel, E. Plass, M. Borges, W. Francke, Journal of Chemical Ecology, 26, 2737-2746 (2000)), Acetonitrile (25 ml). Heated to +40° C. for 5 days. Removed the acetonitrile in vacuo and added diethyl ether (50 ml) to the residue. Yield: 1.2 g (58%).
[0288] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N-dimethyltetradecan-1-aminium bromide (S44):
[0289] Promethazine (1.1 g; 3.7 mmol); 1-Bromotetradecane (0.91 g; 3.3 mmol); Acetonitrile (25 ml). Heated to +40° C. for 7 days. Removed the acetonitrile in vacuo and added petroleum ether Bp<50° C. (50 ml) to the residue. Yield: 1.2 g (60%).
[0290] N-(1-(10H-phenothiazin-10-yl)propan-2-yl)-N,N,3,7-tetramethyloct-6-en-1-aminium 4-methylbenzenesulfonate (S45):
[0291] Promethazine (0.97 g; 3.4 mmol); 3,7-Dimethyloct-6-en-1-yl 4-methylbenzenesulfonate (1.2 g; 3.8 mmol) (synthesized by the procedure of P. H. G. Zarbin, A. Rreckziegel, E. Plass, M. Borges, W. Francke, Journal of Chemical Ecology, 26, 2737-2746 (2000)), Acetonitrile (25 ml). Heated to +40° C. for 7 days. Removed the acetonitrile in vacuo and added diethyl ether (50 ml) to the residue. Yield: 1.7 g (83%).
[0292] N-(2-(10H-phenothiazin-10-yl)ethyl)-N,N-dimethyldodecan-1-aminium bromide (S46):
[0293] N,N-Dimethyl-2-(10H-phenothiazin-10-yl)ethan-1-amine (1.0 g; 3.69 mmol) (synthesized by the procedure of M. Blaess, N. Bibak, R. A. Claus, M. Kohl, G. A. Bonaterra, R. Kinscherf, S. Laufer, H.-P. Deigner, European Journal of Medicinal Chemistry 153, 73-104 (2018)), 1-bromododecane (1.0 g; 4.1 mmol), acetonitrile (25 ml). Heated to +50° C. for 5 days. Removed acetonitrile in a nitrogen stream and added diethylether to the residue. Yield: 1.4 g (75%).
TABLE-US-00001 TABLE 1 Molecular Compound CAS Molecular weight No. Structure Chemical name No. formula CLog P* [g/mol] S1
TABLE-US-00002 TABLE 2 Molecular Compound CAS Molecular weight No. Structure Chemical name No. formula CLog P* [g/mol] S12
TABLE-US-00003 TABLE 3 Com- Molecular pound Molecular weight No. Structure Chemical name CAS No. formula CLog P* [g/mol] S23
TABLE-US-00004 TABLE 4 Molecular Compound Molecular weight No. Structure Chemical name CAS No. formula CLog P* [g/mol] S34
TABLE-US-00005 TABLE 5 Molecu- Com- lar pound CAS Molecular weight No. Structure Chemical name No. formula CLog P* [g/mol] S40
Example 3
[0294] The aim of the study was to determine the Minimal Inhibitory Concentration (MIC) of four original compounds (thioridazine hydrochlorid, promethazine hydrochlorid, chlorpromazine hydrochlorid and chlorprothixene hydrochlorid) and 36 derivatives hereof, for ten different bacterial strains.
[0295] Ten different bacterial strains, representing 8 different bacteria species, were included in the strain collection. For all strains, a MIC value for each compound was established.
TABLE-US-00006 TABLE 6 Bacterial strains for which MIC values were determined for all 38 compound. Strain Genus Species name Gram Ref. Comments Staphylococcus aureus JE2 + [1] MRSA strain Staphylococcus aureus CC398 + MRSA strain Bacillus cereus ATCC_11778 + America-type culture collection Enterococcus faecium Un-named + Enterococcus faecalis Un-named + Staphylococcus pseudintermedius DK729 + Streptocococcus equi JTR204 + Listeria monocytogenes Un-named + Escherichia coli APEC_O2 − [2] Salmonella enteritidis Fagtype 6 − [1] Baek K T, Thogersen L, Mogenssen R G, Mellergaard M, Thomsen L E, Petersen A, Skov S, Cameron D R, Peleg A Y, Frees D. Stepwise decrease in daptomycin susceptibility in clinical Staphylococcus aureus isolates asso ciated with an initial mutation in rpoB and a compensatory inactivation of the clpX gene. Antimicrob Agents Chemother. 2015; 59: 6983-6991. [2] Jorgensen S L, Kudirkiene E, Li L, Christensen J P, Olsen J E, Nolan L, Olsen R H. Chromosomal features of Escherichia coli serotype O2:K2, an avian pathogenic E. coli. Stand Genomic Sci. 2017; 12: 33.
TABLE-US-00007 TABLE 7 Bacterial strains for which MIC values were determined for chlorpromazine hydrochloride and dodecyl. Strain Genus Species name Gram Ref. Comments Escherichia coli ESBL1 − [3] Tetracycline resistant Escherichia coli ESBL10 − [3] Tetracycline resistant Escherichia coli ESBL19 − [3] Tetracycline resistant Escherichia coli ESBL28 − [3] Tetracycline resistant Escherichia coli ESBL37 − [3] Tetracycline resistant Escherichia coli ESBL46 − [3] Tetracycline resistant Escherichia coli E2 − [4] ESBL.sup.1 Escherichia coli E4 − [4] ESBL Escherichia coli E5 − [4] ESBL Escherichia coli E6 − [4] ESBL Escherichia coli E24 − [4] ESBL Escherichia coli E38 − [4] ESBL .sup.1Extended spectrum beta-lactamase [3] Olsen R H, Bisgaard M, Lohren U, Robineau B, Christensen H. Extended-spectrum beta-lactamase-producing Escherichia coli isolated from poultry: a review of current problems, illustrated with some laboratory findings. Avian Pathol. 2014; 43: 199-208. [4] Ahmad A, Zachariasen C, Christiansen L E, Graesboll K, Toft N, Matthews L, Damborg P, Agerso Y, Olsen J E, Nielsen S S. Pharmacodynamic modelling of in vitro activity of tetracycline against a representative, naturally occurring population of porcine Escherichia coli. Acta Vet Scand. 2015; 57: 79.
TABLE-US-00008 TABLE 7b Bacterial strains for which MIC and MBC values were determined for S18 and S43. Strain Genus Species name Gram Ref Cutibacterium acnes X10 + Clinical, multiresistent isolate Staphylococcus Epidermidis RP62A + Clinical, multiresistent isolate Campylobacter jejuni B31C6 − Clinical isolate
[0296] Method:
[0297] The MIC for each compound for each bacteria strain was determined by the serial dilution method following the CLSI guidelines (CLSI Clinical and Laboratory Standards Institute: Performance Standards for Antimicrobial Susceptibility Testing: Twenty-First Informational Supplement M100-S25. Wayne, Pa., USA; 2015.) All compounds were diluted in autoclaved H.sub.2O and stored at 4° C. For each compound, the concentration tested ranged from 0.0125 to 256 mg/L by two-fold increase. Bacterial inoculums were prepared by inoculating 9 ml of H.sub.2O with bacterial colonies from agar plate (Oxoid, Roskilde Denmark) supplemented with bovine blood to a final yield a final density of 10.sup.8 colony forming unit (CFU)/mL using a Sensititre™ Nephelometer (Thermo Scientific, Roskilde, Denmark). These inoculums were each in diluted 1:100 in Müller Hinton (MH) broth (Sigma, Copenhagen, Denmark). From the diluted inoculums 100 μl was transferred to each well in a 96-well plate and mixed with the compound to be MIC determined. The plate was incubated for 24 hours at 37° C. (without shaking). The MIC value was determined as the lowest concentration in which no bacterial growth could be visually detected. All MIC determinations were done in duplicates. If growth was observed in the wells contained 256 mg/L, the MIC value was set to 256 mg/L although the true value may be significantly higher. Since all derivatives are new, no control strains (with reference MIC values) were available.
[0298] The results are presented in table 8, 9 and 10 and in
TABLE-US-00009 TABLE 8 Thioridazine Promethazine Chlorpromazine Chlorprothixene S. aureus USA300 Hydrochloride 32 128 64 32 Methyl 16 128 32 256 Ethyl 2 128 16 32 1-Isopentyl 2 16 16 4 1-Pentyl 2 8 64 4 Isopropyl 8 64 16 1-Dodecyl 4 0.5 0.5 4 3-Ethoxy-3- 32 64 oxo-1-propyl 6-Methoxy-6- 64 8 oxo-1-hexyl Benzyl 2 16 8 4 2-Propene-1-yl 4 64 16 16 3,7-dimethyloct- 4 6-en-1-yl 1-Tetradecyl 8/16 3,7-dimethyloct- 2 (Promazine) 6-en-1-yl E. faecalis Hydrochloride 32 128 64 32 Methyl 128 256 128 256 Ethyl 8 256 128 128 1-Isopentyl 4 64 32 16 1-Pentyl 4 32 128 16 Isopropyl 32 128 128 1-Dodecyl 4 0.5 0.5 3-Ethoxy-3- 32 64 oxo-1-propyl 6-Methoxyy-6- 256 128 oxo-1-hexyl Benzyl 8 128 16 16 2-Propene-1-yl 16 256 64 128 E. coli APEC O2 Hydrochloride 128 128 64 64 Methyl 256 256 256 256 Ethyl 256 256 256 256 1-Isopentyl 128 64 128 64 1-Pentyl 64 64 256 64 Isopropyl 128 128 128 1-Dodecyl 64 64 4 3-Ethoxy-3- 256 128 oxo-1-propyl 6-Methoxy-6- 256 128 oxo-1-hexyl Benzyl 128 128 64 64 2-Propene-1-yl 256 256 256 128 Staphylococus pseudintermedius Hydrochloride 16 64 32 32 Methyl 8 64 8 256 Ethyl 2 32 8 32 1-Isopentyl 2 8 8 4 1-Pentyl 4 4 32 2 Isopropyl 8 32 8 1-Dodecyl 2 0.25 0.25 3-Ethoxy-3- 16 32 oxo-1-propyl 6-Methoxy-6- 32 8 oxo-1-hexyl Benzyl 2 16 2 2 2-Propene-1-yl 4 64 8 8 Streptococcus equi Hydrochloride 16 64 16 256 Methyl 8 32 16 256 Ethyl 2 32 16 256 1-Isopentyl 2 8 8 4 1-Pentyl 2 4 32 2 Isopropyl 8 256 32 1-Dodecyl 2 1 1 3-Ethoxy-3- 8 32 oxo-1-propyl 6-Methoxy-6- 32 8 oxo-1-hexyl Benzyl 2 16 2 2 2-Propene-1-yl 4 32 8 8 S. aureus CC398 Hydrochloride 32 128 32 32 Methyl 16 128 32 256 Ethyl 4 64 16 32 Isopentyl 2 16 16 4 Pentyl 2 32 64 4 Isopropyl 8 16 16 Dodecyl 2 0.25 0.25 3-Ethoxy-3- 32 64 oxo-1-propyl 6-Methoxyyl-6- 64 8 oxo-1-hexyl Benzyl 2 16 4 2 2-Propene-1-yl 4 64 16 16 E. faecium Hydrochloride 16 256 32 32 Methyl 64 128 32 256 Ethyl 8 128 32 128 Isopentyl 4 64 32 16 Pentyl 2 16 64 16 Isopropyl 16 128 64 Dodecyl 2 0.25 0.25 3-Ethoxy-3- 32 32 oxo-1-propyl 6-Methoxyyl-6- 128 64 oxo-1-hexyl Benzyl 8 64 16 16 2-Propene-1-yl 16 128 64 64 Salmonella enteritidis Hydrochloride 256 256 64 128 Methyl 128 128 256 256 Ethyl 256 256 256 256 Isopentyl 128 128 128 64 Pentyl 64 64 256 64 Isopropyl 128 128 128 Dodecyl 64 64 16 3-Ethoxy-3- 128 64 oxo-1-propyl 6-Methoxyyl-6- 256 128 oxo-1-hexyl Benzyl 64 128 64 64 2-Propene-1-yl 128 256 128 128 Listeria monocytogenes Hydrochloride 32 128 32 32 Methyl 32 256 64 256 Ethyl 2 32 32 128 Isopentyl 2 32 32 8 Pentyl 2 8 64 8 Isopropyl 8 128 32 Dodecyl 2 0.25 0.25 3-Ethoxy-3- 4 64 oxo-1-propyl 6-Methoxyyl-6- 128 8 oxo-1-hexyl Benzyl 4 32 8 32 2-Propene-1-yl 32 256 32 32 Bacillus cereus Hydrochloride 32 256 64 64 Methyl 16 128 32 128 Ethyl 2 256 32 64 Isopentyl 4 16 64 8 Pentyl 2 8 64 8 Isopropyl 8 128 32 Dodecyl 2 0.25 0.25 3-Ethoxy-3- 4 128 oxo-1-propyl Metyl hexanoate 128 8 Benzyl 2 32 4 16 2-Propene-1-yl 64 256 64 32
TABLE-US-00010 TABLE 9 Strain Chlorpromazine name Hydrochl Dodecyl ESBL1 32/64 4/4 ESBL10 64/64 8/8 ESBL19 32/32 4/4 ESBL28 32/32 4/4 ESBL37 32/32 4/4 ESBL46 32/32 4/8 E2 64/32 8/4 E4 64/64 4/4 E5 32/32 4/4 E6 32/32 4/4 E24 64/32 4/4 E38 64/32 4/4
TABLE-US-00011 TABLE 10 N-(3-(10H- phenothiazin- 10-yl)propyl)- N,N,3,7- tetramethyloct- 6-en-1-aminium 4- Promethazine ethylbenzenesulfonate dodecyl Original strain (S43) mg/L (S18) mg/L of strain Staphylococcus 2 (MIC) 0.25 (MIC) Multiresistant epidermidis 2 (MBC) 0.25 (MBC) clinical isolate Campylobacter 1 (MIC) Not Clinical jejuni 1 (MBC) evaluated isolate Cutibacterium 1 (MIC) 0.5 (MIC) Multiresistant acnes 1 (MBC) 0.5 (MBC) clinical isolate
Example 4
[0299] To assess the permeability of the compounds, the apical to basolateral flux/permeability of different experimental compounds across monolayers of IPEC-J2 MDR1 cells seeded and cultured on Transwell supports was investigated.
[0300] Materials and Methods:
[0301] Materials
[0302] Bovine serum albumin (BSA), Hank's balanced salt solution (HBSS), Fetal bovine serum (FBS), 2-[4-(2-hydroxyethyl) piperazin-1-yl] ethanesulfonic acid (HEPES), Ultima Gold Scintillation Fluid, Transwell® Permeable supports (1.13 cm2, 0.4 μm pore size), Cytotoxicity Detection Kit (LDH).
[0303] Experimental Compounds [0304] Compound 1: 10-(2-(1-methylpiperidin-2-yl)ethyl)-2-(methylthio)-10H-phenothiazine hydrochloride (S23); [0305] Compound 2: 1-ethyl-1-methyl-2-(2-(2-(methylthio)-1-OH-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S25); [0306] Compound 3: 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S27); [0307] Compound 4: 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide (S28).
[0308] The compounds were dissolved in ultra pure water (MilliQ) to a concentration of 1 mM and further diluted in HBSS supplemented with 0.05% BSA and 10 mM HEPES, pH 7.4 to a final concentration of 10 μM.
[0309] For transport experiments, IPEC-J2 cells were seeded onto to permeable supports (T12, Corning cci-3401) and cultured for 15-17 days. Culturemedium was changed every other day in both the apical and basolateral chamber. On the day of transport, the cells were equilibrated to ambient temperatures and the transepithelial electrical resistance (TEER) was measured across the cell monolayers. Subsequently, the cell layers were washed twice with HBSS and the TEER was measured again. The transport experiment was started by replacing the HBSS in the apical chamber with HBSS solutions of the four experimental compounds. Over a period of two hours samples of 100 μL where taken from the basolateral chamber at t=15, 30, 45, 60, 90 and 120 minutes. All samples were stored at −20° C. until analysis by means of HPLC-MS. All four compounds were tested in triplicate. After the transport study was stopped, the cell layers were washed twice with HBSS and the TEER was measured.
[0310] In a parallel transport experiment, .sup.14C-mannitol (0.5 μCi/mL) was added to all four compound solutions. The experiment was performed as described above, with the exceptions that the four compounds were tested in duplicate and that samples were mixed with 2 mL of Ultima Gold Scintillation Fluid and analysed by liquid scintillation counting.
[0311] Lactate Dehydrogenase (LDH) assay
[0312] LDH-release from cell monolayers was measured in samples of the donor solutions taken from the apical compartment after 120 minutes of exposure on IPEC-J2 MDR1 cell monolayers. Cell lysate of cells treated with ultrasound was added to the analysis as a positive control. Samples were analysed according to manufacturer's protocol in duplicate.
[0313] Quantification of Experimental Compounds
[0314] Lc-Specific:
[0315] Column: Unknown. Phenomenex EVO C18. Column temp: 40° C. Mobilphases: A: MilliQ+0.1% Formic acid; B: Acetonitrile+0.1% Formic acid. Flow: 0.5 mL/min. Injection: 5 uL. UV Detection: 254 nm.
[0316] Gradient:
TABLE-US-00012 Time (minutes) Solvent B (%) 0 20 0.5 20 2 40 8 65 14 20
[0317] Runtime 15 min.
[0318] MS-specific:
[0319] Ionization Mode: MM-ESI. Polarity: Positive. Spray Chamber having Gas temp of 250° C., Vaporizer of 200° C., Drying gas of 12 L/min and Neb. pres. at 35 psig. VCap: 4000 V.
[0320] Corona: 4 A.
TABLE-US-00013 SIM Ion Fragmentor 371.2 70 Compound 1 399.2 70 Compound 2 441.3 70 Compound 3 441.2 70 Compound 4
[0321] Results:
[0322] Apical to Basolateral Transport Across IPEC-J2 MDR1 Cells.
[0323]
[0324]
[0325] Mannitol Transport
[0326]
[0327] LDH Assay
[0328]
[0329] So, the apical to basolateral transport experiment with 10 μM solutions of the four compounds showed that only Compound 1 was able to permeate through the cell monolayer. The appearance of Compound 2, 3, and 4 in the basolateral chamber was barely detectable. Increased mannitol transport was observed for one replicate treated with supplemented HBSS and for one replicate treated with 10 μM solution of Compound 1, which could indicate a disruption of the barrier properties. However, this was not reflected in the LDH release assay, which showed no negative effect of HBSS or the drug solutions on the integrity of the cell membranes.
[0330] Overall the findings of these experiments show that Compound 1 diffuses through IPEC-J2 MDR1 cell monolayers with a high permeability, while the permeability of Compound 2, 3 and 4 is low.
Example 5
[0331] This experiment is conducted to assess the ability of four experimental compounds to lyse red blood cells in vitro. Since quaternary ammonium compounds tends to be toxic due to lysis, the inventors have tested the compounds against red blood cells.
[0332] Materials and Methods:
[0333] Materials
[0334] Bovine whole blood supplemented with citrate, Ultrapure water (MilliQ), Phosphate buffered Saline, 0.1% TritonX in PBS and Isotonic saline.
[0335] Experimental Compounds [0336] Compound TH: 10-(2-(1-methylpiperidin-2-yl)ethyl)-2-(methylthio)-10H-phenothiazine hydrochloride (S23); [0337] Compound TE: 1-ethyl-1-methyl-2-(2-(2-(methylthio)-1-OH-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S25); [0338] Compound TI: 1-isopentyl-1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)piperidin-1-ium bromide (S27); [0339] Compound TP: 1-methyl-2-(2-(2-(methylthio)-10H-phenothiazin-10-yl)ethyl)-1-pentylpiperidin-1-ium bromide (S28).
[0340] The experimental compounds were dissolved in PBS to an initial concentration of 80 mg/L and treated with ultrasound. The initial stock solution was further diluted in PBS to produce solutions with concentrations of 1, 2 and 4 mg/L.
[0341] Red Blood Cell Lysis Assay
[0342] To investigate the ability of drug solutions to lyse red blood cells, 300 μL bovine whole blood was mixed with 1200 μl sample solution. The mixture of whole blood and sample solution was agitated for 30 minutes by end-over-end rotation at ambient temperatures.
[0343] Subsequently, the mixture was centrifuged for 10 minutes at 13000 RPM. The supernatant was diluted 20-fold in ultrapure water and the absorbance of the diluted supernatant at 540 nm was measured (LabSystems Multiscanner plate reader). All samples were measured in duplicate. PBS and isotonic saline was included as negative controls where no lysis of red blood cells is expected. Ultrapure water and a solution of 0.1% TritonX in PBS was included as positive controls where complete lysis of red blood cells is expected.
[0344] Results:
[0345] The results of the red blood cell lysis assay is shown in
[0346] None of the experimental compounds, at concentrations of 1 mg/L, 2 mg/L and 4 mg/L, caused any increase in measured absorbance relative to phosphate buffered saline or isotonic saline (negative controls). It can therefore be concluded, that the tested compounds do not cause lysis of red blood cells in concentrations up to 4 mg/L.
Example 6
[0347] This experiment is conducted to assess in vivo toxicity of the compounds. Compound S43 was selected for this study to evaluate for the highest tolerable dosage in mice. The study was performed at Statens Serum Institute. The Maximum Tolerated Dose (MTD) was investigated after intravenous (iv), intraperitonalt (ip) and peroral (po) dosing ranging from 5 mg/kg to 100 mg/kg. Mice were observed for clinical signs of discomfort for 4-6 hrs after injection. Compound S43 was tolerated after iv and ip dosing up to 10 mg/kg. Mice dosed twice po with 100 mg/kg showed no signs of discomfort as was thus considered well tolerated.
Example 7
[0348] This experiment is conducted to assess the effect of topical treatment of the compounds against MRSA. Compound S43 was evaluated for efficacy against Staphylococcus aureus MRSA 43484 in an murine skin infection model. Treatment with compound S43 formulations at 1% and 2% resulted in significant reduction of the bacterial loads compared to vehicle treatment in the skin lesion.
[0349] Treatment was performed twice daily for 3 days and sampling of skin biopsies was performed the day after last treatment. Treatment with Fucidic acid was included as a positive control and treatment with vehicle was included as a negative control.
[0350] Materials and Methods
[0351] 48 Balb/C female mice—18-22 g—Taconic, Denmark, Staphylococcus aureus MRSA43484, Test compound A at 1 and 2% formulations, Test compound B at 2% formulation, Fucidin, 0.9% saline (sterile, SSI), 0.9% saline/Triton-x (sterile, SSI), Sterile water (SSI), 5% Horse Blood Agar plates (SSI), MRSA Brilliance agar plates, Nurofen® Junior (20 mg/ml, Novartis), Zoletil mix/Torbugesic (SSI/QC-BIO), Dermal curette—Miltex 335710.
[0352] Laboratory Animal Facilities and Housing of Mice:
[0353] The temperature and humidity were registered daily in the animal facilities. The temperature was 22° C.+/−2° C. and can be regulated by heating and cooling. The humidity was 55+/−10%. The air changes per hour were approximately 8-12 times (70-73 times per hours inside racks), and light/dark period was in 12-hours interval of 6 a.m.-6 p.m./6 p.m-6 a.m. The mice had free access to domestic quality drinking water and food (Teklad Global diet 2916C-Envigo) and occasionally peanuts and sunflower seeds (Koge Korn A/S). The mice were housed in Type 3 macrolone cages with bedding from Tapvei. Further, the animals were offered Enviro-Dri nesting material and cardboard houses (Bio-serv).
[0354] Preparation of inoculum: Fresh overnight colonies from a 5% Horse Blood Agar plate were suspended in saline to approximately 109 CFU/ml.
[0355] Preparation of anaesthetic (Zoletil mix): Zoletil mix was diluted before use (total of 10 ml): 2 ml Zoletil mix+5.2 ml sterile saline+2.8 ml Torbugesic 1:100
[0356] Inoculation of Mice, Day 0
[0357] Approximately 1 hour before inoculation, mice were treated orally with 45 μl nurofen (20 mg ibuprofen/ml corresponding to approximately 30 mg/kg) as pain relief. The mice were anaesthetized with 0.15 ml s.c. of Zoletil mix. The fur was removed from a 2×3-cm area on the back of each mouse by use of an electric shaver. Next, a razor was used to remove all the hair and hereafter the outer most layer of the skin was scraped off with a dermal curette to obtain a 1 cm.sup.2 superficial skin lesion. 10 μl inoculum containing approximately 107 CFU was spread on the lesion. After the applied inoculum had dried, the mouse was placed in the cage and kept in a warming cabinet until fully awake.
[0358] Dermal Treatment of Mice, Day 1-3
[0359] Topical treatment of was initiated the day after inoculation, Day 1. Mice were treated twice daily (9 a.m and 3 p.m.) for three days. A volume of 50 μl was spread on the inoculated skin area.
[0360] The mice were observed all days during study and scored 0-4 based on their behaviour and clinical signs.
TABLE-US-00014 Score 0 Healthy Score 1 Minor clinical signs of infection (slower movements) Score 2 Moderate signs of infection (lack of curiosity or changed activity) Score 3 Severe signs of infection (reduced movements, lightly pinched eyes) Score 4 Severe signs of infection (stiff movements, pinched eyes, cold,). Sacrificed.
[0361] Sampling, Day 1 and Day 4
[0362] Colony counts in the skin lesions were determined on day 1 (start of treatment) and day 4 (the day after completed treatment). The mice were sacrificed day 1 and 4 according to Table 1. The affected skin area was removed by a pair of scissors and tweezers, and collected in a tube for Dispomixer with 1 ml saline. The skin sample was homogenized in a Dispomixer. Each sample was serial diluted in saline/Triton-x and 20-μl spots were applied on MRSA Brillince agar plates. All agar plates were incubated 20-48 hrs at 35° C.
[0363] Treatment Schedule.
TABLE-US-00015 Day 1 Day 4 Day 1-3 sampling sampling Day 0 Treatment of skin of skin Inoculation 0.05 ml/mouse lesions lesions Compound B 2% 1-2-3-4- 5-6-7-8 MRSA Compound A 2% 9-10-11-12- 13-14-15-16 Compound A 1% 17-18-19-20- 21-22-23-24 Fucidic acid 25-26-27-28- 29-30-31-32 Vehicle 33-34-35-36- 37-38-39-40 Start of treatment 41-42-43-44- 45-46-47-48
[0364] The colony count in the inoculum was determined to 8.76 log.sub.10 CFU/ml, corresponding to 6.76 log.sub.10 CFU/mouse. Colony counts in skin lesion were performed at day 1 (start of treatment) and day 4 post inoculation. The CFU counts are shown in
[0365] Treatment with 1 and 2% test compound formulations resulted in a significant (p<0.0001; Anova; multiple comparisons) reduction of the bacterial loads of 3.0 and 3.7 log.sub.10 CFU respectively compared to vehicle treatment in the skin lesion. Treatment with fucidic acid 2%, resulted in a 1.5 logo reduction of the CFU levels compared to the vehicle control (p<0.05).
[0366] A carry over effect was observed when counting colonies in spots of 10 fold dilutions of the skin samples. This indicates that a high concentration of active compound was still present in the skin biopsies at the time of sampling. This may in turn underestimate the number of viable bacteria detected on the agar plates for the samples where counting was performed in 10-100 fold dilutions.
Example 8
[0367] The compound S43 was also assessed for mutagenicity. The results showed that S43 did not show indications of mutagenic potential (data not shown).
[0368] The invention is further described in the following non-limiting items.
[0369] Items [0370] 1. A composition comprising a compound of formula I
##STR00044## [0371] wherein [0372] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl; [0373] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl; [0374] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0375] d is selected from 0, 1, 2, and 3; [0376] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0377] e is selected from 0, 1, 2, 3, and 4; [0378] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl; [0379] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3); [0380] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where; [0381] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl; [0382] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl; [0383] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl;
[0384] where A is selected from any pharmaceutical relevant/acceptable anion/counterion;
[0385] wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl;
[0386] wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl. [0387] 2. A composition according to item 1 wherein the compound of formula I is a phenothiazine derivative. [0388] 3. A composition according to item 2 wherein the phenothiazine derivative is selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof. [0389] 4. A composition according to item 1 wherein the compound of formula I is a chlorprothixene derivative. [0390] 5. A composition according to item 3 wherein the compound of formula I is a thioridazine derivative. [0391] 6. A composition according to item 3 wherein the compound of formula I is a promethazine derivative. [0392] 7. A composition according to item 3 wherein the compound of formula I is a Chlorpromazine derivative. [0393] 8. A composition according to any of the preceding items wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl. [0394] 9. A composition according to any of the preceding items wherein Y.sup.3 is selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. [0395] 10. A composition according to any of the preceding items 1 to 7 wherein Y.sup.3 is a linear or branched C.sub.8-15-alkyl. [0396] 11. A composition according to any of the preceding items 1 to 7 or 10 wherein Y.sup.3 is selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl. [0397] 12. A composition according to any of the preceding items wherein Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. [0398] 13. A composition according to any of the preceding items wherein Y.sup.1 and Y.sup.2 are both C.sub.1-alkyl. [0399] 14. A composition according to any of the preceding items 1 to 9 wherein Y.sup.1 together with a carbon being part of the W and the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl. [0400] 15. A composition according to item 12 wherein Y.sup.2 is selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl. [0401] 16. A composition according to any of the preceding items wherein X is S. [0402] 17. A composition according to any of the preceding items wherein Z is selected from the group consisting of hydrogen, Cl or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl. [0403] 18. An anti-microbial composition comprising a compound of formula I
##STR00045## [0404] wherein [0405] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl; [0406] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12-alkyl; [0407] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0408] d is selected from 0, 1, 2, and 3; [0409] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0410] e is selected from 0, 1, 2, 3, and 4; [0411] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl; [0412] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3); [0413] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where; [0414] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl; [0415] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl; [0416] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl; [0417] where A is selected from any pharmaceutical relevant/acceptable anion/counterion; for use as a medicament. [0418] 19. An anti-microbial composition for use according to item 16 wherein the compound of formula I is a phenothiazine derivative. [0419] 20. An anti-microbial composition for use according to item 17 wherein the phenothiazine derivative is selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof. [0420] 21. An anti-microbial composition for use according to item 17 wherein the compound of formula I is a chlorprothixene derivative. [0421] 22. An anti-microbial composition for use according to item 18 wherein the compound of formula I is a thioridazine derivative. [0422] 23. An anti-microbial composition for use according to item 18 wherein the compound of formula I is a promethazine derivative. [0423] 24. An anti-microbial composition for use according to item 18 wherein the compound of formula I is a Chlorpromazine derivative. [0424] 25. An anti-microbial composition for use according to any of the preceding items 18 to 24 wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl. [0425] 26. An anti-microbial composition for use according to any of the preceding items 18 to 25 wherein Y.sup.3 is selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. [0426] 27. An anti-microbial composition for use according to any of the preceding items 18 to 24 wherein Y.sup.3 is a linear or branched C.sub.8-15-alkyl. [0427] 28. An anti-microbial composition for use according to any of the preceding items 18 to 24 or 27 wherein Y.sup.3 is selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl. [0428] 29. An anti-microbial composition for use according to any of the preceding items 18 to 28 wherein Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. [0429] 30. An anti-microbial composition for use according to any of the preceding items 18 to 29 wherein Y.sup.1 and Y.sup.2 are both C.sub.1-alkyl. [0430] 31. An anti-microbial composition for use according to any of the preceding items 18 to 28 wherein Y.sup.1 together with a carbon being part of the Wand the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl. [0431] 32. An anti-microbial composition for use according to item 31 wherein Y.sup.2 is selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl. [0432] 33. An anti-microbial composition for use to any of the preceding items 18 to 32 wherein X is S. [0433] 34. An anti-microbial composition for use according to any of the preceding items 18 to 33 wherein Z is selected from the group consisting of hydrogen, Cl or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl. [0434] 35. An anti-microbial composition for use according to any of the preceding items 18 to 34 where the medicament is having antibacterial properties against resistant strains of bacteria. [0435] 36. An anti-microbial composition for use according to item 35 where the resistant strains (resistant towards conventional antimicrobial) of bacteria is selected from Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria, Escherichia or Salmonella. [0436] 37. An anti-microbial composition for use according to any of the preceding items 18 to 36 where the medicament is having antibacterial properties against Gram-positive bacteria. [0437] 38. An anti-microbial composition for use according to item 37 wherein the Gram-positive bacteria is selected from Staphylococcus, Bacillus, Enterococcus, Streptococcus or Listeria. [0438] 39. An anti-microbial composition for use according to item 38 wherein the Gram-positive bacteria is selected from Staphylococcus aureus, Bacillus cereus, Enterococcus faecium, Enterococcus faecalis, Staphylococcus pseudintermedius, Streptococcus equi, Listeria monocytogenes. [0439] 40. An anti-microbial composition for use according to any of the preceding items 18 to 39 where the medicament is having antibacterial properties against multi-resistant strains and/or antimicrobial sensitive strains of bacteria. [0440] 41. An anti-microbial composition for use according to item 40 where the multi-resistant strains of bacteria is selected from Staphylococcus aureus or Bacillus cereus. [0441] 42. An anti-microbial composition for use according to any one of items 18 to 41 where the medicament has a minimum inhibitory concentration (MIC) below 16 μg/mL, such as below 8 μg/mL, such as below 4 μg/mL or such as below 2 μg/mL. [0442] 43. An anti-microbial composition for use according to any one of items 18 to 42 wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl; and/or wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl. [0443] 44. An anti-microbial composition comprising a compound of formula I
##STR00046## [0444] wherein [0445] X is selected from the group consisting of S, Se, P, PO, SO, NR.sup.1, CR.sup.1, CR.sup.1R.sup.1 or C.sub.0-2-alkyl; [0446] Z is selected from the group consisting of hydrogen, a halogen, SR.sup.4, OR.sup.4, COR.sup.4 where R.sup.4 is a C.sub.1-12alkyl; [0447] each R.sup.2 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0448] d is selected from 0, 1, 2, and 3; [0449] each R.sup.3 is independently selected from the group consisting of C.sub.1-6-alkyl, halogen, C.sub.3-8-cycloalkyl, OH, NH.sub.2, NHR.sup.1, N(R.sup.1).sub.2, O—C.sub.1-6-alkyl, O—C.sub.3-8-cycloalkyl, NH—C.sub.1-6-alkyl, NH—C.sub.3-8-cycloalkyl, S—C.sub.1-6-alkyl, S—C.sub.3-8-cycloalkyl, aryl, heteroaryl, aryloxy, heteroaryloxy, arylamino, heteroarylamino, arylalkyl, heteroarylalkyl, arylalkyloxy and heteroarylalkyloxy; [0450] e is selected from 0, 1, 2, 3, and 4; [0451] R.sup.1 is selected from the group consisting of C.sub.1-6-alkyl, C.sub.3-8-cycloalkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl; [0452] R.sup.5 is N—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3) or C═CH—(CHW)—N(Y.sup.1)(Y.sup.2)(Y.sup.3); [0453] each W is individually selected from the group consisting of linear or branched C.sub.1-6-alkyl or together with the nitrogen atom —N(Y.sup.1)(Y.sup.2)(Y.sup.3)— to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl together with Y.sup.1 where; [0454] Y.sup.1 is selected from the group consisting of C.sub.1-12-alkyl or together with the W and the nitrogen atom to which it is attached forms an optionally substituted nitrogen-containing heteroaryl or optionally substituted nitrogen-containing heterocyclyl; [0455] Y.sup.2 is selected from the group consisting of C.sub.1-12-alkyl; [0456] Y.sup.3 is selected from the group consisting of linear or branched C.sub.2-25-alkyl, linear or branched C.sub.2-25 alkenyl or linear or branched C.sub.2-25 alkynyl; [0457] where A is selected from any pharmaceutical relevant/acceptable anion/counterion; for use in treating a microbial infection in a human subject. [0458] 45. An anti-microbial composition for use according to item 44 wherein the compound of formula I is a phenothiazine derivative. [0459] 46. An anti-microbial composition for use according to item 45 wherein the phenothiazine derivative is selected from the group consisting of chlorpromazine derivatives, promethazine derivatives and thioridazine derivatives, and salts thereof. [0460] 47. An anti-microbial composition for use according to item 45 wherein the compound of formula I is a chlorprothixene derivative. [0461] 48. An anti-microbial composition for use according to item 46 wherein the compound of formula I is a thioridazine derivative. [0462] 49. An anti-microbial composition for use according to item 46 wherein the compound of formula I is a promethazine derivative. [0463] 50. An anti-microbial composition for use according to item 46 wherein the compound of formula I is a Chlorpromazine derivative. [0464] 51. An anti-microbial composition according to any of the preceding items 44 to 50 wherein Y.sup.3 is a linear or branched C.sub.2-6-alkyl. [0465] 52. An anti-microbial composition for use according to any of the preceding items 44 to 51 wherein Y.sup.3 is selected from the group consisting of ethyl, propyl, methyl-butyl, iso-propyl or pentyl. [0466] 53. An anti-microbial composition for use according to any of the preceding items 44 to 50 wherein Y.sup.3 is a linear or branched C.sub.8-15-alkyl. [0467] 54. An anti-microbial composition for use according to any of the preceding items 44 to 50 or 53 wherein Y.sup.3 is selected from the group consisting of linear or branched C.sub.8-alkyl, linear or branched C.sub.10-alkyl, linear or branched C.sub.12-alkyl, linear or branched C.sub.14-alkyl, linear or branched C.sub.15-alkyl. [0468] 55. An anti-microbial composition for use according to any of the preceding items 44 to 54 wherein Y.sup.1 and Y.sup.2 are individually selected from C.sub.1-6-alkyl, such as from C.sub.1-3-alkyl. [0469] 56. An anti-microbial composition for use according to any of the preceding items 44 to 55 wherein Y.sup.1 and Y.sup.2 are both C.sub.1-alkyl. [0470] 57. An anti-microbial composition for use according to any of the preceding items 44 to 54 wherein Y.sup.1 together with a carbon being part of the Wand the nitrogen atom to which it is attached forms a six-membered nitrogen-containing heterocyclyl. [0471] 58. An anti-microbial composition for use according to item 57 wherein Y.sup.2 is selected from C.sub.1-3-alkyl, such as C.sub.1-alkyl. [0472] 59. An anti-microbial composition for use to any of the preceding items 44 to 58 wherein X is S. [0473] 60. An anti-microbial composition for use according to any of the preceding items 44 to 59 wherein Z is selected from the group consisting of hydrogen, Cl or a SR.sup.4 where R.sup.4 is a C.sub.1-alkyl. [0474] 61. An anti-microbial composition for use according to any of the preceding items 44 to 60 where the medicament is having antibacterial properties against resistant strains of bacteria. [0475] 62. An anti-microbial composition for use according to item 60 where the resistant strains (resistant towards conventional antimicrobial) of bacteria is selected from Staphylococcus, Bacillus, Enterococcus, Streptococcus, Listeria, Escherichia or Salmonella. [0476] 63. An anti-microbial composition for use according to any of the preceding items 44 to 62 where the medicament is having antibacterial properties against Gram-positive bacteria. [0477] 64. An anti-microbial composition for use according to item 63 wherein the Gram-positive bacteria is selected from Staphylococcus, Bacillus, Enterococcus, Streptococcus or Listeria. [0478] 65. An anti-microbial composition for use according to item 64 wherein the Gram-positive bacteria is selected from Staphylococcus aureus, Bacillus cereus, Enterococcus faecium, Enterococcus faecalis, Staphylococcus pseudintermedius, Streptococcus equi, Listeria monocytogenes. [0479] 66. An anti-microbial composition for use according to any of the preceding items 44 to 65 where the medicament is having antibacterial properties against multi-resistant strains of bacteria. [0480] 67. An anti-microbial composition for use according to item 66 where the multi-resistant strains of bacteria is selected from Staphylococcus aureus or Bacillus cereus. [0481] 68. An anti-microbial composition for use according to any one of items 44 to 67 where the medicament has a minimum inhibitory concentration (MIC) below 16 μg/mL, such as below 8 μg/mL, such as below 4 μg/mL or such as below 2 μg/mL. [0482] 69. An anti-microbial composition for use according to any one of items 44 to 68 wherein if X is S and Z is a halogen then Y.sup.3 cannot be a C.sub.2-alkyl or a branched C.sub.3-alkyl; and/or wherein if X is S and Z is hydrogen then Y.sup.3 cannot be C.sub.2-alkyl or linear or branched C.sub.5-alkyl.