CA2265765A1 - New amino alcohol derivatives, process for the production thereof and medicaments and reagents containing these compounds - Google Patents

Abstract

The present invention relates to new amino alcohol derivatives, process for the production thereof and medicaments and reagents containing these compounds. The subject of the invention is medicaments having the general formula (I) wherein A can mean hydrogen, a group NR1R2, a group NR1(CH2)pNR3R4, a group (C=NH)NH2 or a pyridinyl radical, B and D can be identical or different and stand each for a binding, a C1 to C6-alkylene radical or a group NR5-C2-C6-alkylene, C can mean piperidindiyl or piperazindiyl, W and X are identical or different and stand each for a carbonyl group, Y and Z can be identical or different and stand each for a saturated or unsaturated hydrocarbon radical with 7-24 carbon atoms, R1 to R5 can be identical or different and stand each for hydrogen or a C1 to C6 alkyl radical, m stands for a whole number 0, 1, or 2 and if m = 2 both radicals C
independently from another can also mean piperidindiyl and piperazindiyl, n and o can be identical or different and stand each for a whole number 2, 3 or 4 and p means a whole number from 2 to 6, as well as their physiologically compatible salts, with the proviso that no hydrazine derivatives are comprised and that m be not 0, when A means hydrogen or a group (C=NH)NH2 and B and D
are identical or different and represent a binding or an alkylene radical.

Description

CA 02265765 l999-03- 102New amino alcohol derivatives, process for theirproduction and pharmaceutical preparations and reagentscontaining these compoundsThe present invention concerns new amino alcoholderivatives, a process for their production as well aspharmaceutical preparations and reagents which containthese substances.The invention concerns pharmaceutical agents of thegeneral formula I(CH2)n-O-W-YA—B-Cm-D-N\(CH2)o-O-X-Z (I),in whichA denotes hydrogen, a group NR1R2, a groupNR1(CH2)PNR3R4, a group (C=NH)NH2 or a pyridinylresidue,B and D are the same or different and each denotes abond, a C1 to C6 alkylene residue or a groupNR5-C2 to C6 alkylene,C denotes piperidinediyl or piperazinediyl,-... ... ................-.m......u-s................ ...... .,CA 02265765 l999-03- 10W and X are the same or different and each denotes abond or a carbonyl group,4-—Y and Z are the same or different and each denotes asaturated or unsaturated hydrocarbon residuewith 7 to 24 carbon atoms,R1 to R5are the same or different and each representshydrogen or a C1 to C6 alkyl residue,m is an integer 0, 1 or 2 and if m equals 2 bothresidues C can be the same or different,n and o are the same or different and each denotes theintegers 2, 3 or 4 andp denotes an integer from 2 to 6as well as physiologically tolerated salts thereof,provided that hydrazine derivatives are not included andthat m cannot be 0 if A denotes hydrogen or a group(C=NH)NH2 and B and D being the same or differentrepresent a bond or an alkylene residue.The alkyl, alkylene and hydrocarbon residues encompassedby the meanings of B, D, Y, Z and R1 to R5 can bestraight—chained or branched. A pyridinyl residue isunderstood as an unsubstituted pyridine or a pyridineoptionally substituted severa1—fo1d with straight-chained or branched C1-C6.CA 02265765 l999-03- 10Values of 1 or 2 are preferred for m. Compounds of thegeneral formula I are just as preferred which containmore than 2 nitrogens or, if Aidenotes (C=NH)NH2, thosethat contain more than 3 nitrogens.The invention in addition concerns new amino alcoholderivatives of the general formula I(CH2)n-O-W-YA—B-Cm-D-N(CH2)o - O - X - Z (I),in whichA denotes hydrogen, a group NR1R2, a groupNR1(CH2)pNR3R4, a group (C=NH)NH2 or a pyridinylresidue,B and D are the same or different and each denotes abond, a C1 to C6 alkylene residue or a groupNR5-C2 to C6 alkylene,C denotes piperidinediyl or piperazinediyl,W and X are the same or different and each denotes abond or a carbonyl group,Y and Z are the same or different and each denotes asaturated or unsaturated hydrocarbon residuewith 7 to 24 carbon atoms,R1 to R5 are the same or different and each representsCA 02265765 l999-03- 10hydrogen or a C1 to C6 alkyl residue,m is an integer 0, 1 or 2’and if m equals 2 bothresidues C can be the same or different,n and o are the same or different and each denotes theintegers 2, 3 or 4 andp denotes an integer from 2 to 6as well as physiologically tolerated salts thereof,provided that hydrazine derivatives are not included andthat m cannot be 0 if A denotes hydrogen or a group(C=NH)NH2 and B and D being the same or differentrepresent a bond or an alkylene residue and that it doesnot include the compoundsoctadecanoic acid-[(3-diethylamino-propyl)imino]-bis(methyl-2,1-ethanediyl)—ester-hydrochlorideoctadecanoic acid—[(3—dimethylamino-propyl)imino]di—3,1-propanediyl ester-dihydrochlorideoctadecanoic acid-[(3—dimethylamino—propyl)imino]di—3,1-propanediyl esteroctadecanoic acid-[(3-dimethylamino-propyl)imino]di-2,1-ethanediyl ester-hydrochloridedocosanoic acid-2—[(3-dimethylamino-propyl)—[2-[(1-oxododecyl)oxy]ethyl]amino]ethyl esterhexadecanoic acid-[[2—(ethylmethylamino)ethyl]imino]di-2,1-ethanediyl esteroctadecanoic acid-[[3-(dimethylamino)propyl]imino]di-2,1-ethanediyl esteroctadecanoic acid-[[3-(dimethylamino)propyl]imino]di—2,1-ethanediyl ester—dihydrochlorideoctadecanoic acid-[[3-(dimethylamino)propyl]imino]di—CA 02265765 l999-03- 102,1-ethanediyl esterN,N-Bis[3-(dodecyloxy)propyl]-1,2-ethanediamineoctadecanoic acid—[[2-[(2-aminoethyl)amino]ethyl]imino]—di-2,1-ethanediyl esterstearic acid—iminobis-(ethyleneiminoethylene)-ester-monoacetate.Within the sense of the present invention the followingmeanings in compounds of formula I are preferredindependently of one another. This applies to compoundsas well as to medicaments that contain such compoundsand likewise also to the respective therapeuticapplications of these compounds.A denotes NH2 or N(CH3)2 orB and D are the same or different and denote abond, a C1-C3 alkylene residue or if m=0 anN(CH3)C3 to C4 alkylene residue orC denotes piperidinediyl orm denotes 0 or 1 orW and X each denote CO orY denotes Cl3H27 or C17H33 orZ denotes C13H27 or C17H33 orn and o each denote 2.Compounds are especially preferred which fulfil all theabove-mentioned meanings simultaneously.The compounds of formula I have valuable pharmacologicalproperties and in particular they can facilitate thetransport of biologically active molecules intoprokaryotic or eukaryotic cells. They are thereforeparticularly suitable for introducing proteins, nucleicacids such as e.g. DNA, cDNA, mRNA, PNA, antisenseCA 02265765 l999-03- 10polynucleotides and therapeutically active low molecularcompounds such as peptide hormones, cytostatic agentsand antibiotics into target cells within or outside ofthe organism. The new compounds according to theinvention are therefore particularly suitable for theefficient treatment of mammals by gene therapypreferably of human patients. These compounds are alsosuitable for the production of drug combinations incancer therapy, antiviral therapy, infection therapy andin diseases caused by dysregulation. In contrast toviral carriers for gene constructs, non-viral geneferries often have only a low immunogenicity. Theefficiency and persistency of the gene expressionmediated by non—viral gene ferries has, however, not yetbeen satisfactory. In addition to improving geneexpression, the compounds of the general formula I havethe advantage that they can be degraded relativelyeasily due to the C2-C4—alkyl-0 chains on the tertiarynitrogen N of formula I.Apart from the compounds listed in the examples, theinvention concerns in particular all substances whichhave all possible combinations of the meanings of thevariables mentioned in the examples.The process according to the invention for theproduction of compounds of formula I is characterized inthat a compound of the general formula II(CH2)n-O-HA-B—Cm-D-N(CH2)o ‘ 0 “ H (II).in which A, B, C, D, m, n and o have the above-mentionedCA 02265765 l999-03- 10meaning is reacted with a compound of the generalformula III and a compound of the general formula IVE - W - Y G - X - Z(III) (IV)in which W, X, Y and Z have the above-mentioned meaningand E and G represent reactive residues,and subsequently if desired a protecting group containedin A, B or D is cleaved—off, a hydrogen atomrepresenting A is converted into a group (C=NH)NH2, acompound present as an acid addition salt is convertedinto the free base or a compound present as a base isconverted by neutralization with a non-toxic acid into aphysiologically tolerated salt.The reactive residues E and G are nucleofuge groups suchas for example halogen atoms, sulfonate or sulfategroups or acidic residues of activated esters,anhydrides or mixed anhydrides.It is expedient to react compounds of formula II withcompounds of formulae III and IV in an inert solventsuch as an ether, for example tetrahydrofuran or anamide such as dimethylformamide or in pyridineoptionally in the presence of a base such astriethylamine or ethyldiisopropylamine or an alkalialcoholate, however, the reagents of the generalformulae III or IV can be used undiluted or, if W and Xrepresent a carbonyl group, an acid such as acetic acidor trifluoroacetic acid can be used as the solvent.CA 02265765 l999-03- 10Cleavage of a protecting group contained in A, B or D iscarried out depending on the chemical characteristics ofthis group, for example by acidic or basic hydrolysis orhydrogenolysis. An acid cleavable protecting group isfor example the tert.-butoxycarbonyl residue.A hydrogen atom representing A can for example beconverted into an amidino group by reaction withcyanamide or pyrazol—1—carboxamidine.The majority of the starting compounds of the generalformula II are new (especially if m represents 1 or 2)and are also a subject matter of the invention. They canbe produced from known starting materials by methodsknown in the literature.Potential pharmacologically acceptable salts are inparticular salts with non-toxic inorganic or organicacids such as for example hydrochloric acid, sulfuricacid, phosphoric acid, hydrobromic acid, acetic acid,trifluoroacetic acid, lactic acid, citric acid, malicacid, benzoic acid, salicylic acid, malonic acid, maleicacid, succinic acid or diaminocaproic acid.The salts are obtained in the usual manner for exampleby neutralizing the compounds of formula I with thecorresponding acids.In order to produce pharmaceutical preparations ortransfer reagents the compounds according to theinvention are combined individually or as a combination,if desired using co-lipids, with a biologically activemolecule, for example a polynucleotide, in a suitableratio and administered in vivo or in vitro in a liquidpreferably aqueous, or solid preferably lyophilizedCA 02265765 l999-03- 10form. The in vivo administration can be carried outorally, parenterally, topically, transmucosally or byintroduction into a body cavity-of the patient. Adelayed release from a biologically degradable matrix oradministration as an aerosol or inhalable powderapplication is also possible.The administered dose depends on the age, health andweight of the recipient, the extent of the disease, thetype of other treatments which may be carried out at thesame time, the frequency of the treatments and the typeof the desired effect and can be determinedexperimentally by a person skilled in the art.The following compounds are preferred within the senseof the invention in addition to the substances mentionedin the examples:1. Oleic acid-2-[(2-oleoyloxy—ethy1)-piperidin-4-yl-methyl)-amino]-ethyl ester2. Tetradecanoic acid-2—[(2-tetradecanoyloxy-ethyl)-piperidin—4-yl—methyl)-amino]-ethyl ester3. Dodecanoic acid-2-[(2-dodecanoyloxy-ethyl)-piperidin-4-yl—methyl)-amino]-ethyl ester4. Oleic acid-2-[(2—oleoyloxy-ethy1)-[1,4']bipiperidinyl—4-yl-amino]-ethyl ester5. Tetradecanoic acid—2-{(2-tetradecanoyloxy-ethyl)-[3-[1,4']bipiperidinyl-4-ylamino)—propyl]-amino}-ethyl ester..s.....................u. .~......-..._. 10.ll.12.13.14.CA 02265765 l999-03- 10-10..Dodecanoic acid-2-{(2—dodecanoyloxy-ethyl)—[3-([1,4']bipiperidinyl-4-ylamino)-propyl]—amino}-rethyl ester -Oleic acid—2-{(2—oleoyloxy-ethyl)-[3~(3,4,5,6-tetrahydro-2H-[1,4']bipyridiny1-4-ylamino)—propyl]—amino}—ethy1 esterDodecanoic acid—2—[{3-[3—dimethylamino—propyl)—methyl—amino]—propyl}—(2-dodecanoyloxy-ethyl)-amino]—ethyl esterTetradecanoic acid-2—[{3-[(3—dimethylamino-propyl)—methyl—amino]-propyl}-(2-tetradecanoyloxy—ethy1)-amino]—ethy1 esterDodecanoic acid-2—[{3-[(3-diethylamino-propyl)—methyl—amino]—propyl}-(2-dodecanoyloxy-ethyl)-amino]-ethyl esterTetradecanoic acid-2—[{3-[(3—diethylamino—propy1)—methyl—amino]-propyl}-(2-tetradecanoyloxy-ethyl)-amino]-ethyl esterDodecanoic acid-2-{[3—(4-amino-butylamino)-propyl]-(2-dodecanoyloxy-ethyl)-amino}—ethyl esterDodecanoic acid-2-{(2-dodecanoyloxy-ethyl)-[1-(2-amino-ethyl)—piperidin—4-yl]—amino}-ethyl esterOleic acid—2-{(2-oleoyloxy-ethyl)-[1-(2-amino-ethyl)-piperidin—4-yl]-amino}—ethyl ester15.16.17.18.19.20.21.22.23.24.CA 02265765 l999-03- 10-11..Dodecanoic acid-2—{(2-dodecanoyloxy-ethyl)-[1—(2-amino-ethyl)-piperidin-4-yl-methyl]—amino}—ethy1ester ,-Oleic acid—2-{(2-oleoyloxy—ethy1)—[l-(2-amino-ethyl)-piperidin-4—yl-methyl]-amino}—ethyl esterDodecanoic acid—2-{[2~(4-amino-piperidin-1—y1)-ethyl]-(2-dodecanoyloxy-ethyl)-amino}-ethyl esterDodecanoic acid-2-{[4-(4-amino-piperidin—1-yl)—butyl]-(2-dodecanoyloxy-ethyl)—amino}—ethy1 esterTetradecanoic acid-2-{[2-(4-amino-piperidin-1-yl)-ethyl]—(2—tetradecanoyloxy-ethyl)—amino}-ethylesterTetradecanoic acid—2-{[4-(4-amino—piperidin-1-yl)-butyl]—(2—tetradecanoy1oxy-ethyl)-amino}-ethylesterOleic acid—2-{[2-(4—amino—piperidin-1-yl)—ethyl]—(2—oleoy1oxy—ethyl)—amino}-ethyl esterOleic acid—2—{[4-(4—amino—piperidin-1-yl)-butyl]-(2-oleoyloxy-ethyl)—amino}-ethyl esterOleic acid-2-{(2-oleoyloxy-ethyl)-[1-(2—amino—ethyl)-piperidin-4-yl]—amino}—ethyl esterOleic acid-2-{(2-oleoyloxy-ethyl)-[l-(3-amino-propyl)-piperidin-4-yl—methyl]—amino}—ethyl ester. .,-........._....—._._.CA 02265765 l999-03- 1025. Oleic acid-2—{(2—oleoyloxy-ethyl)-[1-(2-amino-ethyl)-piperidin-4-yl—methyl]—amino}-ethyl esterf26. 4-Dimethylamino-1—{3-[bis-(2-tetradecyloxy-ethyl)-amino]—propyl}-piperidine27. 4'—{[Bis-(2—tetradecanoyloxy-ethyl)]-amino}-[4,1']bipiperidine-1-carboxamidineExample 1Decanoic acid-2—{(2-decanoyloxy-ethyl)-[3—(4-methyl-piperazin—1-yl)-propyl]-amino}—ethyl ester-hydrochloride2.1 ml (15 mmol) triethylamine is added to a solution of1.35 g (5.5 mmol) 2—{(2-hydroxy-ethyl)-[3-(4-methyl-piperazin-1-yl)-prpyl]-amino}—ethanol in 30 mltetrahydrofuran, a solution of 2.3 ml (11 mmol) decanoylchloride in 20 ml tetrahydrofuran is added dropwise andit is heated for 20 h to reflux. After cooling it isfiltered, the filtrate is concentrated by evaporationand the residue is chromatographed on silica gel. 1.28 gof the desired compound is eluted as an oil with ethylacetate/methanol 1:1, this is dissolved in ethyl acetateand admixed with excess ethereal hydrogen chloridesolution. After concentrating the solution byevaporation the precipitate is removed by filtration and1.3 g of the title compound (42 % of theory) of meltingpoint 208-212°C is isolated.The 2-{(2-hydroxy—ethyl)-[3-(4-methyl-piperazin-1-yl)—propyl]-amino}-ethanol used as the starting material canbe obtained as follows:CA 02265765 l999-03- 10-13-50 mg potassium iodide is added to a solution of 4.1 g(40 mmol) diethanolamine and 7.8 g (44 mmol) 3—(4—methyl-piperazin-1-yl)-propyl.chloride in 40 mldimethyl-formamide and heated for 5 h to 60°C. Afterconcentrating in a vacuum, the residue ischromatographed on silica gel. 2.7 g (27 % of theory) ofthe desired compound is eluted as an oil using ethylacetate/methanol 1:1.Example 2Oleic acid-2—{(2-oleoyloxy-ethyl)—[3-(4—methyl-piperazin-1—yl)-propyl]-amino}-ethyl ester-hydrochlorideThe title compound is obtained as an oil in an analogousmanner to that described in example 1 from 2-{(2-hydroxy-ethyl)-[3-(4-methyl-piperazin-1-yl)-propy1]—amino}-ethanol and oleoyl chloride in a yield of 35 %.Example 3Oleic acid-2-[(2-oleoyloxy-ethyl)-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl—4-yl-amino]—ethyl ester-hydrochlorideThe title compound is obtained as a viscous oil in ananalogous manner to that described in example 1 from2-hydroxyethyl-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl)-amino]-ethanol and oleoyl chloride in a yield of53o\°The 2—hydroxy-ethyl-(3,4,5,6-tetrahydro—2H—[1,4']bipyridinyl-4—yl)-amino]-ethanol used as theCA 02265765 l999-03- 10-14-starting material can be obtained as follows:a) A mixture of 62.9 g (0.33 mol) 1-benzyl-piperidin-4-one, 34.8 g diethanolamine and 300 ml toluene is heatedfor 2 h to reflux on a water separator. After 5 ml waterhas been separated, it is concentrated and the residueis distilled in a vacuum. 73.2 g 2-(8-benzyl-1-oxa-4,8-diaza-spiro[4.5]-dec-4—yl)-ethanol (80 % of theory) ofb.p.o_o8 185-190°C is isolated.b) A solution of 87.6 g (0.32 mol) of the previouslydescribed compound in 900 ml methanol is hydrogenatedover 2 g platinum dioxide and subsequently over 2 g10 percent palladium carbon at 4 bar hydrogen pressure.After the calculated amount of hydrogen has been taken55.0 g (91 % oftheory) 2-[(2-hydroxy-ethyl)-(piperidin—4-yl)-amino]-up, it is filtered and concentrated.ethanol remain as an oil.c) A mixture of 24 g (127 mmol) of the previouslydescribed compound and 7.2 g (64 mmol) 4—chloro-pyridineis heated for 2 h to 150°C, subsequently taken up in10 N sodium hydroxide solution and extracted withdichloro methane and methanol. After concentrating theextract and triturating with ethyl acetate, 14.3 g (91 %of theory) 2-[2-hydroxy-ethyl-(3,4,5,6-tetrahydro-2H-[1,4'jbipyridinyl-4—y1)—amino]—ethanol of m.p. 126—128°Cremain.Example 4Tetradecanoic acid-2—[(2-tetradecanoyloxy—ethyl)-(3,4,5,6-tetrahydro-2H—[1,4'jbipyridinyl-4-yl)-amino]-ethyl ester—hydrochlorideCA 02265765 l999-03- 10-15-The title compound is obtained in an analogous manner tothat described in example 1 in a 51 % yield as anamorphous powder of melting range 100—120°C from 2-[(2-hydroxy-ethyl)-(3,4,5,6—tetrahydro—2H-[1,4']bipyridinyl—4-yl)-amino]—ethanol and tetradecanoyl chloride.Example 5Dodecanoic acid~2-[(2—dodecanoyloxy-ethyl)-3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl)-amino]—ethylester-hydrochlorideThe title compound is obtained as an amorphous powder inan analogous manner to that described in example 1 from2-[(2—hydroxy-ethyl)—(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl—4-yl)-amino]—ethanol and dodecanoylchloride in a yield of 48 %.Example 6Oleic acid—2-[(2-oleoyloxy-ethyl)-(3,4,5,6—tetrahydro—2H-[1,4']bipyridinyl-4-yl-methyl)-amino]-ethyl ester-hydrochlorideThe title compound is obtained as an oil in an analogousmanner to that described in example 1 from 2-[(2-hydroxy-ethyl)-(3,4,5,6—tetrahydro-2H-[1,4']bipyridinyl-4—yl-methyl)-amino]-ethanol and oleoyl chloride in ayield of 37 %.The 2—hydroxyethyl—(3,4,5,6—tetrahydro-2H-[1,4']bipyridinyl-4-yl)—amino]-ethanol used as thestarting material can be obtained as follows:CA 02265765 l999-03- 10-16-a) A mixture of 23.8 g (0.1 mol) 1—benzoyl-4-chloromethyl-piperidine and 21.0 g (0.1 mol)diethanolamine is heated for l,h to 150°C, subsequently10 N sodium hydroxide solution is added and it isextracted with dichloromethane. After concentration byevaporation, 30.4 g (99 % of theory) 2-[(2-hydroxy-ethyl)—(1-benzoyl-piperidin—4-yl-methyl)-amino]-ethanolremain as an oil.b) 30.3 g of the previously described compound is heatedwith 200 ml 6 N hydrochloric acid for 5 h to reflux.After cooling it is washed with diethyl ether, theaqueous phase is concentrated, it is made alkaline with10 N sodium hydroxide solution, extracted with dichloro-methane, dried and concentrated by evaporation. 16.0 g(79 % of theory) 2-[(2-hydroxy—ethyl)—(piperidin-4-yl-methyl)-amino]-ethanol remain as an oil.c) 2-[(2-Hydroxy-ethyl)-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4—yl-methyl)-amino]-ethanol isobtained as an oil analogously to the process describedunder 3c) from the previous compound and 4 chloro-pyridine in a 44 % yield.Example 7Tetradecanoic acid-2—[(2-tetradecanoy1oxy~ethy1)—(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl-methyl)-amino]-ethyl ester-hydrochlorideThe title compound is obtained as an oil in an analogousmanner to that described in example 1 from 2-[(2-hydroxy—ethyl)—(3,4,5,6—tetrahydro-2H-[l,4']bipyridinyl—4—yl-methyl)—amino]-ethanol and tetradecanoyl chlorideVA ..l..._..........-—....-........CA 02265765 l999-03- l0_l7._in an 86 % yield.Example 8 ’Dodecanoic acid-2—[(2-dodecanoyloxy-ethyl)-(3,4,5,6—tetrahydro-2H—[1,4']bipyridinyl-4-yl-methyl)-amino]-ethyl ester-hydrochlorideThe title compound is obtained as an oil in an analogousmanner to that described in example 1 from 2-[(2-hydroxy-ethyl)-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl—4-yl-methyl)—amino]-ethanol and dodecanoyl chloride inan 81 % yield.Example 9Oleic acid—2-[(2—oleoyloxy-ethyl)-(3—piperidin-1-yl-propyl)-amino]-ethyl esterThe title compound is obtained as an oil in an analogousmanner to that described in example 1 from 2-[(2-hydroxy-ethyl)—(3-piperidin-1—yl-propyl)-amino]-ethanol(J. Organomet. Chem. gfil, 289 (1983)) and oleoylchloride in a 39 % yield.Example 10Dodecanoic acid-2—{[3-(4-amino-piperidin-1-yl)-propyl]-2—dodecanoy1oxy-ethyl)-amino}-ethyl ester1.0 g (5 mmol) dodecanoyl chloride is added dropwise toa solution of 0.7 g (2 mmol) 2—{[3—(4-amino-piperidin-1-CA 02265765 l999-03- 10-18-yl)-propyl]—2—hydroxy-ethyl)-amino}-ethanol-hydrochloride in 10 ml trifluoroacetic acid and it isstirred for 18 h at room temperature. It is concentratedby evaporation, admixed with 10 ml cold 1 N sodiumhydroxide solution, extracted with ethyl acetate,concentrated by evaporation and chromatographed onsilica gel. 0.87 g (71 % of theory) of the desiredcompound is eluted as an oil with ethyl acetate/methanol4:1.The 2-{[3-(4-amino-piperidin-1—yl)-propyl]-(2-hydroxy-ethyl)-amino}-ethanol-hydrochloride used as the startingmaterial can be obtained as follows:a) A mixture of 56.2 g (0.2 mol) 4-benzamido-1-(3-chloro—propyl)—piperidine, 19.8 g (0.19 mol)diethanolamine, 16.6 g (0.12 mol) potassium carbonateand 500 ml n-propanol are heated for 32 h to reflux. Itis filtered, the filtrate is concentrated by evaporationand chromatographed on silica gel. 33.2 g (48 % oftheory) 2-{[3-(4-benzamido-piperidin—1-yl)-propyl]-(2-hydroxy—ethyl)—amino}-ethanol is eluted with ethylacetate/methanol 9:1 as an amorphous solid substance.b) 17.2 g (0.05 mol) of the previous compound isrefluxed for 18 h with 190 ml 6 N hydrochloric acid. Itis allowed to cool, washed with dichloromethane and theaqueous phase is concentrated by evaporation. 17.0 g(96 % of theory) 2-{[3-(4-amino-piperidin—1-yl)-propy1]-(2-hydroxy—ethy1)—amino}-ethanol-hydrochloride remain asan oil.CA 02265765 l999-03- 10-19..Example 11Tetradecanoic acid-2—{[3-(4-amino-piperidin-1-yl)-propyl]—(2-tetradecanoyloxy—ethyl)—amino}—ethyl esterThe title compound is obtained as an oil in a 64 % yieldin an analogous manner to that described in example 10from 2—{[3-(4-amino-piperidin-1—yl)-propyl—(2-hydroxy—ethyl)-amino}-ethanol-hydrochloride and tetradecanoylchloride.Example 12Oleic acid-2—[(2-oleoyloxy—ethyl)-(2-piperidin-4-yl-ethyl)—amino]-ethyl ester-hydrochloride3.1 ml (9.5 mmol) oleoyl chloride is added to a solutionof 0.95 g (3.8 mmol) 2-[(2-hydroxy-ethyl)-(2-piperidin—4—yl-ethyl)-amino]-ethanol—hydrochloride in 20 mldimethylformamide and heated for 4 h to 50°C. It isconcentrated by evaporation and chromatographed onsilica gel. 0.4 g (14 % of theory) of the title compoundis eluted as an oil with ethyl acetate/methanol 9:1.The 2—[(2—hydroxy—ethyl)-(2-piperidin—4-yl—ethyl)-amino]-ethanol used as a starting material can beobtained as follows:1.0 g activated ruthenium oxide is added to a solutionof 57 g (0.27 mol) 2-[(2-hydroxy-ethyl)-(2-pyridin-4—yl-ethyl)-amino]-ethanol (Chem. Abstr. iggg, 13129) in700 ml methanol and hydrogenated for 16 h at 100°C and150 bar hydrogen pressure. It is filtered, concentrated. ‘.._.................“................._......-_ ..... 41CA 02265765 l999-03- 10-20-by evaporation and chromatographed on silica gel. 47.9 g(82 % of theory) of the desired compound is eluted as anoil with methanol. ,—Example 13Tetradecanoic acid-2-[(2-tetradecanoyloxy-ethyl)—(2-piperidin-4-yl-ethyl)-amino]-ethyl esterThe title compound is obtained as an oil in a 71 % yieldin an analogous manner to that described in example 10from 2—[(2-hydroxy-ethyl)-(2-piperidin-4—yl-ethyl)-amino]-ethanol and tetradecanoyl chloride.Example 14Dodecanoic acid-2—[(2-dodecanoyloxy~ethyl)-(2—piperidin-4-yl-ethyl)-amino]—ethyl esterThe title compound is obtained as an oil in a 90 % yieldin an analogous manner to that described in example 10from 2-[(2—hydroxy—ethyl)-(2-piperidin-4—yl-ethyl)—amino]-ethanol and dodecanoyl chloride.Example 15Tetradecanoic acid-2—{(2—tetradecanoyloxy-ethyl)—[2-(3,4,5,6-tetrahydro—2H—[1,4']bipyridinyl-4—yl)—ethyl]—amino}-ethyl esterThe title compound is obtained as an oil in a 95 % yieldin an analogous manner to that described in example 10CA 02265765 l999-03- 10-21-from 2—{(2-hydroxy-ethyl)-[2-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl)—ethyl]-amino}—ethano1 andtetradecanoyl chloride. ,-Example 16Dodecanoic acid-2-{(2-dodecanoyloxy—ethyl)-[2-(3,4,5,6—tetrahydro-2H-[1,4']bipyridinyl—4—yl)—ethyl]-amino}-ethyl esterThe title compound is obtained as an oil in a 65 % yieldin an analogous manner to that described in example 10from 2-{(2-hydroxy-ethyl)-[2-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl)—ethy1]-amino}—ethanol anddodecanoyl chloride.Example 17Tetradecanoic acid—2-[(2-tetradecanoyloxy-ethyl)-[l,4']bipiperidinyl-4-yl—amino]—ethyl esterThe title compound is obtained as an oil in a 35 % yieldin an analogous manner to that described in example 10from 2-[(2-hydroxy—ethyl)—[1,4']bipiperidinyl-4-yl-amino]-ethanol and tetradecanoyl chloride.The 2-[(2—hydroxy-ethyl)-[1,4']bipiperidinyl-4—y1-amino]-ethanol used as a starting material can beobtained as follows:24 g (90 mmol) of the 2-[2—hydroxy-ethyl—(3,4,5,6-tetrahydro-2H—[1,4']bipyridiny1—4-yl)-amino]-ethanoldescribed under 3c) is hydrogenated on ruthenium oxideCA 02265765 l999-03- 10-22....analogously to the process described in example 12. 17 g(70 % of theory) of the desired compound is isolated asan oil. ,-Example 18Dodecanoic acid-2-[(2-dodecanoyloxy—ethyl)-[1,4']bipiperidinyl-4—yl—amino]-ethyl esterThe title compound is obtained as an oil in a 32% yieldin an analogous manner to that described in example 10from 2-[(2-hydroxy—ethyl)-[1,4']bipiperidinyl-4—yl—amino]-ethanol and dodecanoyl chloride.Example 19Oleic acid-2-{(2-oleoyloxy-ethyl)—[2—(3,4,5,6-tetrahydro-2H-[1,4'jbipyridinyl-4-yl)-ethyl]-amino}-ethyl ester-hydrochlorideThe title compound is obtained as an oil in a 57% yieldin an analogous manner to that described in example 1from 2-{(2—hydroxy-ethyl)-[2-(3,4,5,6-tetrahydro-2H-[1,4']bipyridinyl-4-yl)—ethyl]-amino}-ethanol and oleoylchloride.Example 20Oleic acid-2-[(2-oleoyloxy-ethyl)-(1‘-methyl-[1,4']bipiperidiny1—4—y1)-amino]—ethyl esterThe title compound is obtained as an oil in a 37 % yield .-........-..........................u...... ..CA 02265765 l999-03- 10-23-in an analogous manner to that described in example 1 from2-[(2—hydroxy-ethyl)-(1'-methy1—[1,4']bipiperidinyl—4—yl)-amino]-ethanol and oleoyl chloride.The 2-[(2-hydroxy-ethyl)—(1'-methyl-[1,4']bipiperidinyl-4-yl)-amino]-ethanol used as a starting material can beobtained as follows:a) 5.4 g (20 mmol) of the 2—[(2-hydroxy-ethyl)-[l,4']bipiperidinyl-4-yl—amino]-ethanol described inexample 17 is refluxed for 5 h with 20 ml ethyl formateand 1 ml water and subsequently concentrated byevaporation. 6.2 g (quantitative) 2-[(2-hydroxy—ethyl)-(1'-formyl-[1,4']bipiperidinyl-4-yl)-amino]—ethanolremain as an oil.b) 6.0 g (20 mmol) of the previously described compoundis added dropwise to a suspension of 2.9 g lithiumtetrahydridoaluminate in 150 ml tetrahydrofuran andsubsequently refluxed for 3 h. After standing overnight,ethyl acetate and saturated saline solution are added,it is filtered and the filtrate is concentrated byevaporation. It is chromatographed on silica gel and1.8 g (32 % of theory) 2-[(2-hydroxy-ethyl)-(1'—methy1—[1,4']bipiperidinyl—4-yl)—amino]-ethanol is eluted as anoil with ethyl acetate/methanolic ammonia 9:1.Example 21Dodecanoic acid-2-{(2-dodecanoyloxy-ethyl)-[1-(3-amino-propyl)-piperidin-4-yl]—amino}-ethyl esterThe title compound is obtained as an oil in a 37 % yieldCA 02265765 l999-03- 10-24-in an analogous manner to that described in example 10from 2-{(2-hydroxy-ethyl)-[1—(3-amino—propyl)-piperidin-4-yl]-amino}-ethanol and dodecanoyl chloride.The 2—{(2-hydroxy-ethyl)-[1—(3-amino-propyl)-piperidin—4-yl]-amino}-ethanol used as a starting material can beobtained as follows:a) A mixture of 5.5 g (29 mmol) of the 2—[(2-hydroxy—ethyl)-(piperidin-4—yl)-amino]-ethanol described in 3b),60 ml methanol and 1.9 ml (29 mmol) acrylonitrile isstirred for 24 g at room temperature and subsequentlyconcentrated in a vacuum. 6.7 g (96 % of theory) 2-{(2-hydroxy-ethyl)—[l-(2-cyano-ethyl)-piperidin-4-yl]—amino}-ethanol remain as an oil.b) 6.7 g (28 mmol) of the previously described compoundis hydrogenated over Raney-nickel in 150 ml methanolicammonia at 100°C and 100 bar hydrogen pressure. It isfiltered, concentrated by evaporation and 6.8 g(quantitative) 2-{(2—hydroxy—ethyl)-[1-(3—amino-propyl)—piperidin—4—yl]-amino}-ethanol is obtained as an oil.Example 22Tetradecanoic acid-2—{(2-tetradecanoyloxy-ethyl)-[1—(3-amino—propyl)-piperidin-4-yl]-amino}-ethyl esterThe title compound is obtained as an oil in a 34 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy—ethyl)-[1—(3—amino-propyl)-piperidin-4-yl]—amino}-ethanol and tetradecanoyl chloride.CA 02265765 l999-03- 10-25.-Example 23Dodecanoic acid-2—{(2-dodecanoyloxy-ethyl)-[1—(3-amino-propyl)—piperidin—4—yl-methyl]-amino}-ethyl esterThe title compound is obtained as an oil in a 44 % yieldin an analogous manner to that described in example 10from 2-{(2-hydroxy—ethyl)-[1—(3-amino-propyl)-piperidin—4-yl-methyl]-amino}—ethano1 and dodecanoyl chloride.The starting material used can be obtained from thecompound of example 6b) by reaction with acrylonitrileanalogously to example 21a) and subsequent hydrogenationanalogously to example 21b).Example 24Tetradecanoic acid-2-{(2-tetradecanoyloxy-ethyl)-[1-(3-amino-propyl)—piperidin-4-yl-methyl]-amino}-ethyl esterThe title compound is obtained as an oil in a 38 % yieldin an analogous manner to that described in example 10from 2-{(2—hydroxy-ethyl)—[1—(3-amino—propyl)—piperidin—4-yl-methyl]-amino}-ethanol and tetradecanoyl chloride.Example 25Tetradecanoic acid—2-{(2-tetradecanoy1oxy—ethyl)-[3-(3,4,5,6-tetrahydro—2H-[1,4']bipyridinyl—4—ylamino)-propyl]-amino}—ethyl esterThe title compound is obtained as an oil in a 29 % yieldCA 02265765 l999-03- 10-26-in an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)-[3-(3,4,5,6—tetrahydro-2H-[1,4']bipyridinyl-4-ylamino)-propyl]-amino}—ethanol andtetradecanoyl chloride.The 2—{(2-hydroxy-ethyl)—[3—(3,4,5,6—tetrahydro-2H-[1,4']bipyridiny1-4-ylamino)-propyl]-amino}—ethanol usedas a starting material can be obtained as follows:a) A solution of 46 g (0.4 mol) 4-chloropyridine and123.5 g (0.86 mol) 4—piperidone ethylene ketal isrefluxed for 48 h in 400 ml p-xylene. It is cooled,filtered, the filtrate is concentrated by evaporationand chromatographed on silica gel. 79.7 g (90 % oftheory) 8-pyridin-4-yl—l,4-dioxa-8-aza-spiro[4.5]decanewith a melting point of 65°C is eluted with ethylacetate/ ammoniacal methanol 9:1.b) A solution of 79.7 g of the previously describedketal in 2 l tetrahydrofuran is admixed with 1000 ml 6 Nhydrochloric acid and stirred for 2 h at roomtemperature. It is concentrated by evaporation, madebasic with semi-concentrated ammonia water and extractedwith dichloromethane. After concentrating the extract byevaporation 64.2 g (quantitative) 2,3,5,6—tetrahydro-[1,4']bipyridin-4—one of melting point 102°C remains.c) A mixture of 15 g (85 mmol) of the previouslydescribed ketone, 13.8 g (85 mmol) 2-[3-amino—propyl)-(2-hydroxy-ethyl)-amino]-ethanol (J. Am. Chem. Soc. §§,728 (1944)),toluene is heated for 3 h on a water separator and100 mg 4-toluene sulfonic acid and 200 mlsubsequently concentrated by evaporation. The residue istaken up in 200 ml methanol, 500 mg platinum dioxide isCA 02265765 l999-03- 10-27-.added and it is hydrogenated for 10 h at 1 bar hydrogenpressure. After filtration and concentration byevaporation, it is chromatographed on silica gel and22.4 g 2—{(2-hydroxy-ethyl)-[3-(3,4,5,6—tetrahydro-2H-[1,4']bipyridinyl—4—ylamino)-propyl]—amino}-ethanol iseluted as an oil with ethyl acetate/methanol 1:1.Example 26Dodecanoic acid—2—{(2—dodecanoyloxy-ethyl)-[3-(3,4,5,6-tetrahydro~2H-[1,4']bipyridinyl—4—y1amino)-propyl]—amino}-ethyl ester%The title compound is obtained as an oil in a 34 yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy—ethyl)-[3-(3,4,5,6-tetrahydro—2H—[1,4']bipyridinyl-4-ylamino)-propyl]—amino}-ethanol anddodecanoyl chloride.Example 27Oleic acid—2—{(2-oleoyloxy-ethyl)-[1'-(3-dimethylamino—propyl)-[1,4']bipiperidinyl-4-yl]—amino}-ethyl ester%The title compound is obtained as an oil in a 23 yieldan analogous manner to that described in example 1 from2—{(2-hydroxy—ethyl)-[1'-(3—dimethylamino—propyl)-[1,4']bipiperidinyl-4—yl]—amino}-ethanol and oleoylchloride.The 2-{(2—hydroxy—ethyl)-[1'-(3-dimethylamino-propyl)-[1,4']bipiperidinyl-4-yl]~amino}-ethanol used as thestarting material can be obtained as follows:__.s &..., .CA 02265765 l999-03- 10-28-A mixture of 4.05 g (15 mmol) of the 2-[(2-hydroxy-ethyl)~[1,4']bipiperidinyl-4-yl-amino]-ethanol describedin example 17, 2.0 g potassium,carbonate, 2.2 g 3-dimethylamino-propyl chloride and 25 ml n-propanol isrefluxed for 5 h, cooled, filtered and the filtrated isconcentrated by evaporation. It is chromatographed onsilica gel and 2.0 g (37 % of theory) of the desiredcompound is eluted as an oil with ethylacetate/methanolic ammonia 1:1.Example 28Oleic acid—2—{[3-(4-dimethylamino-piperidin—1-yl)-propyl]-(2-oleoyloxy-ethyl)—amino}—ethyl esterThe title compound is obtained as an oil in a 65 % yieldin an analogous manner to that described in example 1from 2-{[3-(4-dimethylamino—piperidin-1-yl)~propyl]-(2-hydroxy-ethyl)-amino}-ethanol and oleoyl chloride.The 2—{[3-(4-dimethylamino—piperidin-1—yl)—propyl]-(2-hydroxy—ethyl)—amino}—ethanol used as a startingmaterial can be obtained as follows:a) 4-Dimethylamino-piperidine is obtained as an oil in a75 % yield by hydrogenation over ruthenium oxide from 4-dimethylamino-pyridine analogously to the precursordescribed under example 12.b) A mixture of 15.6 g (0.15 mol) diethanolamine, 30.9 g(0.3 mol) 1—bromo—3—chloro-propane, 300 mltetrahydrofuran and 12.6 g potassium carbonate isrefluxed for 5 h, filtered, concentrated by evaporationCA 02265765 l999-03- 10-29-and chromatographed on silica gel. 16.7 g (62 % oftheory) N-(3-chloro-propyl)—diethano1amine is eluted asan oil with ethyl acetate/methanol 9:1.c) A mixture of 1.92 (15 mmol) of the piperidine a),2.72 g (15 mmol) of the halogenide b), 1.2 g potassiumcarbonate and 50 ml n-propanol is refluxed for 6h,filtered, concentrated by evaporation andchromatographed on silica gel. 2.7 g (66 % of theory) 2-{[3-(4-dimethylamino-piperidin-1-yl)-propyl]—(2-hydroxy—ethyl)-amino}-ethanol is eluted as an oil with ethylacetate/methanolic ammonia 1:1.Example 29Oleic acid—2-{(2-oleoyloxy—ethyl)-[1-(3—dimethylamino—propyl)—piperidin-4-yl]-amino}—ethyl esterThe title compound is obtained as an oil in a 24 % yieldin an analogous manner to that described in example 1from 2—{(2-hydroxy-ethyl)-[1-(3-dimethylamino—propyl)—piperidin-4—yl]-amino}-ethanol and oleoyl chloride.The 2-{(2—hydroxy—ethyl)-[1-(3—dimethylamino—propyl)—piperidin—4-yl]-amino}—ethanol used as the startingmaterial can be obtained as follows:A mixture of 5.84 g (30 mmol) of the 2—[(2-hydroxy—ethyl)—(piperidin—4-yl)—amino]—ethanol described under3b), 4.0 g (33 mmol) 3-dimethylamino-propyl chloride,2.5 g potassium carbonate and 20 ml n-propanol isrefluxed for 5 h, filtered, concentrated by evaporationand chromatographed on silica gel. 4.4 g (54 % ofCA 02265765 l999-03- 10-30-theory) of the desired compound is eluted as an oil withethyl acetate/methanolic ammonia 1:1..——Example 30Oleic acid-2-{[3-(4-amino-piperidin-1-yl)-propyl]-(2-oleoyloxy-ethyl)-amino}-ethyl esterA suspension of 4.5 g (4 mmol) of the 2—{[3—(4-amino-piperidin-1—yl)-propyl]-(2-hydroxy-ethyl)-amino}-ethanol—hydrochloride described in example 10 in 100 mldichloromethane is admixed with 3.5 g N—ethyl—diisopropylamine and 0.87 g (4 mmol) pyrocarbonic aciddi-t-butyl ester, it is refluxed for 18 h, a solution of2.4 g (8 mmol) oleoyl chloride in 40 ml dichloromethaneis added dropwise, it is refluxed for 18 h, admixed with15 ml etheric hydrogen chloride solution and stirred for6 h at room temperature. It is concentrated byevaporation, adjusted to pH 9 with N sodium hydroxidesolution, extracted with dichloromethane and methanol,dried and concentrated by evaporation. Afterchromatography on silica gel (eluting agent ethylacetate/methanol 1:1) 1.1 g (36 % of theory) of thetitle compound is isolated as an oil.Example 31Oleic acid-2-{[3-(4-aminomethyl-piperidin-1-yl)—propyl]-(2-oleoyloxy-ethyl)—amino}—ethyl esterThe title compound is obtained as an oil in a 28 % yieldin an analogous manner to that described in example 30from 2-{[3-(4—amino—methyl-piperidin-1—yl)—propyl]—(2-CA 02265765 l999-03- 10-31.-hydroxy-ethyl)-amino}—ethanol and oleoyl chloride.The 2-{[3-(4-aminomethyl—piperidin—1-y1)—propyl]-(2-hydroxy-ethyl)—amino}-ethanol used as the startingmaterial can be obtained from 4—aminomethy1—piperidineanalogously to the compound described under 28c.Example 32Oleic acid-2-[(2—oleoyloxy-ethyl)-(1'-ethyl-[1,4']bipiperidinyl-4-yl)—amino]-ethyl estero\°The title compound is obtained as an oil in a 22 yieldin an analogous manner to that described in example 1from 2-[(2-hydroxy-ethyl)—(1'-ethyl-[1,4']bipiperidinyl-4-yl)—amino]-ethanol and oleoyl chloride.The 2-[(2—hydroxy-ethyl)-(1'-ethyl-[1,4']bipiperidinyl-4-y1)—amino]—ethanol used as the starting material canbe obtained as follows:a) 5.4 g (20 mmol) of the 2-[(2-hydroxy-ethyl)-[1,4']bipiperidinyl—4-yl-amino]—ethanol described inexample 17 is stirred for 5 h at room temperature in20 ml dimethylformamide and 60 ml dichloromethanecontaining 1.6 ml acetyl chloride and 1.7 g sodiumhydrogen carbonate. It is filtered, dried andconcentrated by evaporation. 6.4 g (quantitative) 2-[(2-hydroxy-ethyl)-(1'—acety1-[1,4']bipiperidinyl—4—yl)—amino]—ethanol remains as an oil.b) 5.6 g (18 mmol) of the previously described compoundis reduced analogously to example 20b). 2.1 g (39 % of..... ................._—...—....-....—__....._..~..,.CA 02265765 l999-03- 10-32-theory) 2-[(2-hydroxy-ethyl)-(1'—ethyl-[1,4']bipiperidinyl-4-yl)-amino]-ethanol is obtained asan oil. ,-Example 33Oleic acid-2-{(2—oleoyloxy-ethyl)-[1—(3-amino-propyl)-piperidin-4-yl]-amino}—ethyl esterThe title compound is obtained as an oil in a 25 % yieldin an analogous manner to that described in example 30from 2—{(2—hydroxy—ethyl)—[1—(3-amino-propyl)-piperidin—4-yl]—amino}-ethanol (example 21b) and oleoyl chloride.Example 34Tetradecanoic acid-2-{(2—tetradecanoyloXy-ethyl)—[1-(3-dimethylamino-propyl)—piperidin-4-yl—methyl]—amino}-ethyl esterThe title compound is obtained as an oil in a 31 % yieldin an analogous manner to that described in example 10from 2-{(2-hydroxy-ethyl)-[1-(3-dimethylamino—propyl)-piperidin—4-yl-methyl]-amino}—ethanol and tetradecanoylchloride.The 2-{(2-hydroxy-ethyl)—[1-(3-dimethylamino-propyl)-piperidin-4-yl-methyl]-amino}—ethanol used as thestarting material can be obtained as follows:2-{(2-Hydroxy—ethyl)—[1-(3-dimethylamino-propyl)-piperidin—4-yl—methyl]-amino}—ethano1 is obtained as anoil in a 29 % yield analogously to the process describedCA 02265765 l999-03- 10-33..in example 27 by alkylation of the compound described inexample 6b) with 3-dimethylamino-propyl chloride.rExample 35Oleic acid—2-{(2-oleoyloxy-ethyl)-[1—(3-dimethylamino—propyl)—piperidin-4—yl-methyl]-amino}-ethyl esterThe title compound is obtained as an oil in a 26 % yieldin an analogous manner to that described in example 1from 2—{(2-hydroxy-ethyl)-[1-(3-dimethylamino-propyl)-piperidin-4-yl—methyl]-amino}—ethanol and oleoylchloride.Example 36Tetradecanoic acid-2-{(2-tetradecanoyloxy—ethyl)-2—[[1-(3-amino—propyl)-piperidin—4—yl]-ethyl]—amino}—ethylesterThe title compound is obtained as an oil in a 44 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)-2—[[1-[3-amino-propyl)-piperidin-4—yl]-ethyl]-amino}-ethanol and tetradecanoylchloride.The 2-{(2—hydroxy-ethyl)—2—[[1-(3—amino—propyl)-piperidin-4—yl]-ethyl]—amino}-ethanol used as thestarting material can be obtained analogously to thereaction procedure described in examples 21a) and 21b)from acrylonitrile and the precursor of example 12(yield 47 %).CA 02265765 l999-03- 10_34._Example 37Dodecanoic acid-2—{(2—dodecanoyIoxy-ethyl)-2-[[1-(3-amino-propyl)—piperidin—4-yl]-ethyl]—amino}-ethyl esterThe title compound is obtained as an oil in a 61 % yieldin an analogous manner to that described in example 10from 2-{(2—hydroxy-ethyl)—2—[[1-(3-amino—propyl)-piperidin-4-yl]-ethyl]-amino}—ethanol and dodecanoylchloride.Example 38Tetradecanoic acid-2-{(2—tetradecanoyloxy—ethyl)—2-[[1-(3-dimethylamino-propyl)-piperidin-4—yl]-ethy1]-amino}-ethyl esterThe title compound is obtained as an oil in a 50 % yieldin an analogous manner to that described in example 1from 2-{(2-hydroxy-ethyl)—2-[[1-(3-dimethylamino-propyl)-piperidin-4—yl]-ethyl]-amino}—ethanol andtetradecanoyl chloride.The 2-{(2—hydroxy—ethyl)-2-[[1-(3-dimethylamino—propy1)-piperidin-4—yl]-ethyl]-amino}-ethanol used as thestarting material can be obtained in a 66 % yield fromthe precursor of example 12 and 3—dimethy1amino—propylchloride analogously to the precursor of example 27.Example 39Oleic acid—2-{(2—oleoy1oxy-ethyl)-2—[[1-(3-dimethyl-amino-propyl)-piperidin-4-yl]—ethyl]-amino}—ethyl esterCA 02265765 l999-03- 10_35-OThe title compound is obtained as an oil in a 41 6 yieldin an analogous manner to that described in example 1from 2-{(2-hydroxy—ethyl)—2-[[1:(3—dimethylamino-propyl)-piperidin-4-yl]-ethyl]~amino}-ethanol and oleoylchloride.Example 40Tetradecanoic acid—2—{[3-(4—(2—amino-ethyl)-piperidin—1—yl)—propyl]-(2—tetradecanoyloxy—ethyl)-amino}—ethylester32 % of the theoretical yield of the title compound isobtained as an oil in an analogous manner to thatdescribed in example 10 from 2—{[3-(4-(2-amino-ethyl)-piperidin—1-yl)-propyl]-(2-hydroxy-ethyl)-amino}-ethanoland tetradecanoyl chloride.The 2—{[3-(4-(2~amino-ethyl)-piperidin—1—yl]-propyl]—(2-hydroxy—ethyl)—amino}-ethanol used as the startingmaterial can be obtained as follows:a) Hydrogenation of 4-(2-amino-ethyl)-pyridine (J. Am.Chem. Soc. 78, 4129 (1956)) yields 4-(2-amino—ethyl)-piperidine in a yield of 68 % in an analogous procedureto that described in example 12.b) 9.0 g (70 mmol) of the previous compound is heatedfor 1 h to 150°C with 6.4 g (35 mmol) of the compoundfrom example 28b). It is allowed to cool, taken up in15 ml 2 N sodium hydroxide solution and made stronglyalkaline with 10 N sodium hydroxide solution. Afterextraction with dichloromethane, drying andCA 02265765 l999-03- 10-36-concentrating the extract by evaporation it ischromatographed on silica gel. 6.0 g (63 % of theory)2-{(3—(4-(2-amino—ethyl)—piperidin-1-yl)-propyl]—(2-hydroxy-ethyl)-amino}-ethanol is eluted with ethylacetate/methanol 3:1.Example 41Dodecanoic acid—2-{[3-(4-(2-amino-ethyl)-piperidin-1-yl)-propyl]-(2-dodecanoyloxy-ethyl)-amino}—ethyl esterThe title compound is obtained as an oil in 35 % of thetheoretical yield in an analogous manner to thatdescribed in example 10 from 2-{[3—(4-(2—amino-ethyl)-piperidin-1-yl)-propyl]-(2—hydroxy—ethyl)-amino}-ethanoland dodecanoyl chloride.Example 42Dodecanoic acid—2-{[3-(4-(3-amino-propylamino)—piperidin-1-yl)-propyl]-(2-dodecanoyloxy-ethyl)-amino}-ethyl esterThe title compound is obtained as an oil in 18 % of thetheoretical yield in an analogous manner to thatdescribed in example 10 from 2—{[3-(4—(3—amino-propylamino)-piperidin—l-yl)—propyl]-(2-hydroxy—ethyl)-amino}-ethanol and dodecanoyl chloride.The 2-{[3-(4-(3—amino-propylamino)-piperidin-1-yl)-propyl]-(2-hydroxy—ethy1)-amino}-ethanol used as thestarting material can be obtained as follows:CA 02265765 l999-03- 10...37_Reaction of the 2-{[3-(4—amino—piperidin-1-yl)-propyl]—(2—hydroxy-ethyl)-amino}-ethanol described in example 10with acrylonitrile analogously,to example 21a) andsubsequent hydrogenation analogously to example 21b)yields the desired compound as an oil.Example 43Tetradecanoic acid-2-{[3—(4-(3-amino—propylamino)—piperidin-1-yl)-propyl]-(2-tetradecanoyloxy-ethyl)—amino}-ethyl esterThe title compound is obtained as an oil in 12 % of thetheoretical yield in an analogous manner to thatdescribed in example 10 from 2-{[3—(4-(3—amino-propylamino)—piperidin—1—yl)-propyl]—(2-hydroxy—ethyl)-amino}-ethanol and tetradecanoyl chloride.Example 444-<2—{[Bis-(2—tetradecanoyloxy—ethyl)]-amino}—ethyl>—piperidine-1—carboxamidineA mixture of 1.92 g (3 mmol) of the compound fromexample 13, 0.25 g cyanamide and 5 ml n—butanol isheated for 2 h to 120°C, cooled, the residue is taken upin dichloromethane, washed with a small amount of water,dried and concentrated by evaporation. Afterchromatography on silica gel 0.96 g (47 % of theory) ofthe title compound is eluted as a wax with ethylacetate/methanol 1:1.CA 02265765 l999-03- 10-38-Example 45N-[3—(4-<2[Bis-(2-tetradecanoyI6xy-ethyl)—amino]-ethyl>-piperidin-1—yl)-propyl]-guanidineThe title compound is obtained as an oil in a 64 % yieldin an analogous manner to that described in example 44from the compound of example 36 and cyanamide.Example 46Oleic acid—2—[(2—ethylamino—ethy1)-(2-oleoyloxy-ethyl)—amino]-ethyl ester-hydrochlorideThe title compound is obtained as an oil in a 36 % yieldin an analogous manner to that described in example 1from 2—[(2-ethylamino-ethyl)-(2—hydroxy-ethyl)-amino]-ethanol and oleoyl chloride.The 2—[(2-ethylamino-ethyl)-(2-hydroxy—ethyl)-amino]-ethanol used as the starting material can be obtained asfollows:11.0 g (58 mmol) N-{2—[bis—(2-hydroxy-ethyl)-amino]-ethyl}—acetamide (J. Med. Chem. ;§, 1839 (1993)) isreduced analogously to example 20b). 8.3 g (81 % oftheory) of the desired compound is isolated as an oil.Example 47Oleic acid-2-[(2-diethylamino—ethyl)-(2—oleoyloxy-ethyl)-amino]-ethyl ester-hydrochlorideCA 02265765 l999-03- 10-39-The title compound is obtained as an oil in a 44 % yieldin an analogous manner to that described in example 12from 2-[(2—diethylamino-ethyl);(2-hydroxy-ethyl)—amino]-ethanol and oleoyl chloride.Example 48Oleic acid-2—[(2—amino-ethyl)-(2-oleoyloxy-ethyl)—amino]—ethyl ester-hydrochlorideThe title compound is obtained as an oil in a 25 % yieldin an analogous manner to that described in example 12from 2-[(2-amino—ethyl)—(2—hydroxy-ethyl)-amino]-ethanol(J. Am. Chem. Soc. 81, 3984 (1959)) and oleoyl chloride.Example 49Oleic acid-2-{[3-(3-amino—propylamino)-propyl]-(2-oleoyloxy—ethyl)-amino}-ethyl ester-hydrochlorideThe title compound is obtained as an oil in a 29 % yieldin an analogous manner to that described in example 12from 2—{[3-(3-amino-propylamino)-propyl]-(2-hydroxy-ethyl)-amino}—ethanol and oleoyl chloride.The 2-{[3-(3—amino—propylamino)—propyl]-(2—hydroxy-ethyl)-amino}—ethanol used as the starting material canbe obtained by reaction of 2-[3-amino-propyl)-(2-hydroxy—ethyl)-amino]-ethanol (J. Am. Chem. Soc. §§, 728(1944)) with acrylonitrile analogously to example 21a)and subsequentl hydrogenation analogously to example21b) . b.p.o_o5 176-177°C....... u........-........—.—.——..........._.. .CA 02265765 l999-03- 10-40..Example 50Oleic acid—2—[(3-dimethylamino=propyl)-(2—o1eoyloxy-ethyl)—amino]—ethyl ester-hydrochlorideThe title compound is obtained as an oil in a 65 % yieldin an analogous manner to that described in example 1from 2-[(3-dimethylamino—propy1)-(2-hydroxy—ethyl)-amino]-ethanol and oleoyl chloride.The 2—[(3—dimethylamino-propyl)-(2-hydroxy—ethy1)—amino]-ethanol used as the starting material can beobtained analogously to the procedure of example 27 fromdiethanolamine and 3-dimethylamino-propyl chloride.b.p.1_5 135-136°C.Example 51Oleic acid-2-[(3—diethylamino-propyl)-(2—oleoyloxy-ethyl)-amino]-ethyl esterThe title compound is obtained as an oil in a 65 % yieldin an analogous manner to that described in example 1from 2-[(3-diethylamino-propyl)-(2-hydroxy-ethyl)-amino]-ethanol (Chem. Pharm. Bull. 9, 313 (1961)) andoleoyl chloride.Example 52Tetradecanoic acid-2—[(3—dimethylamino-propyl)-(2-tetradecanoyloxy—ethy1)—amino]-ethyl esterCA 02265765 l999-03- 10-41-The title compound is obtained as an oil in a 78 % yieldin an analogous manner to that described in example 1from 2-[(3—dimethyl—amino—propyl)-(2—hydroxy-ethyl)-amino]-ethanol and tetradecanoyl chloride.Example 53Dodecanoic acid—2—[(3-dimethylamino-propyl)—(2—dodecanoyloxy-ethyl)-amino]-ethyl esterThe title compound is obtained as an oil in a 79 % yieldin an analogous manner to that described in example 1from 2—[(3-dimethylamino-propyl)—(2-hydroxy-ethyl)-amino]—ethanol and dodecanoyl chloride.Example 54Tetradecanoic acid—2-[(3-diethylamino-propyl)—(2-tetradecanoyloxy-ethyl)-amino]-ethyl esterThe title compound is obtained as an oil in a 50 % yieldin an analogous manner to that described in example 1from 2-[(3-diethylamino-propyl)—(2-hydroxy—ethyl)-amino]-ethanol and tetradecanoyl chloride.Example 55Dodecanoic acid—2-[(3-diethylamino-propyl)-(2-dodecanoyloxy-ethyl)-amino]-ethyl estero\°The title compound is obtained as an oil in a 60 yieldin an analogous manner to that described in example 1CA 02265765 l999-03- 10-42.-from 2-[(3—diethylamino-propyl)-(2—hydroxy-ethyl)—amino]-ethanol and dodecanoyl chloride.Example 56Oleic acid-2-[{3—[(3-dimethylamino-propyl)—methyl-amino]—propyl}-(2-oleoyloxy-ethyl)-amino]—ethyl esterThe title compound is obtained as an oil in a 63 % yieldin an analogous manner to that described in example 1from 2-[{3-[(3-dimethylamino-propyl)-methyl-amino]-propyl}—(2-hydroxy-ethyl)-amino]-ethanol and oleoylchloride.The 2-[{3—[(3-dimethylamino-propyl)—methyl-amino]-propyl}-(2-hydroxy-ethyl)-amino]-ethanol used as thestarting material can be obtained analogously to example28c) by alkylation of N,N,N'—trimethyl-propane-1,3-diamine (J. Chem. Soc. (C) iggg, 527) with N-(3-chloro-propyl)-diethanolamine.Example 57Oleic acid-2-[{3-[(3-diethylamino-propyl)-methyl-amino]-propy1}-(2-oleoyloxy-ethyl)-amino]—ethyl esterThe title compound is obtained as an oil in a 59 % yieldin an analogous manner to that described in example 1from 2-[{3—[(3-diethy1amino—propy1)-methyl—amino]—propyl}—(2-hydroxy-ethyl)-amino]-ethanol and oleoylchloride.The 2—[{3—[(3-diethylamino-propyl)-methyl-amino]-CA 02265765 l999-03- 10-43-propyl}—(2-hydroxy—ethyl)-amino]—ethano1 used as thestarting material can be obtained analogously to theprecursor described in example_56 by alkylation of N,N—diethyl-N'-methyl-propane—1,3-diamine ("Monatsh. Chem.11g, 825 (1981)) with N-(3-chloro-propyl)-diethanolamine.Example 58Dodecanoic acid-2-{[4-(3-amino-propylamino)-butyl]-(2-dodecanoyloxy-ethyl)—amino}-ethyl ester-hydrochlorideThe title compound is obtained as a wax in a 31 % yieldin an analogous manner to that described in example 10from 2-{[4—(3-amino-propylamino)-butyl]-(2—hydroxy-ethyl)-amino}-ethanol and dodecanoyl chloride.The 2-{[4-(3-amino-propylamino)—butyl]-(2-hydroxy-ethyl)-amino}-ethanol used as the starting material canbe obtained 2—[(4—amino—butyl)-(2—hydroxy-ethyl)—amino]-ethanol (J. Am. Chem. Soc. §;, 3984 (1959)) and acrylo—nitrile analogously to example 21a) and subsequenthydrogenation analogously to example 21b).Example 59Dodecanoic acid—2—{[3-(3—amino-propylamino)—propy1]—(2-dodecanoyloxy—ethyl)-amino}-ethyl ester-hydrochlorideThe title compound with a melting point of 245-246°C isobtained in a 49 % yield in an analogous manner to thatdescribed in example 10 from 2-{[3-(3—amino—propylamino)-propyl]-(2-hydroxy-ethyl)—amino}—ethanolCA 02265765 l999-03- 10-44-(see example 49) and dodecanoyl chloride.Example 60 "Tetradecanoic acid-2-{[4—(3-amino-propylamino)-butyl]-(2-tetradecanoyloxy-ethyl)-amino}-ethyl esterThe title compound is obtained as an oil in a 27 % yieldin an analogous manner to that described in example 10from 2—{[4-(3—amino—propy1amino)-butyl]-(2-hydroxy-ethyl)—amino}-ethanol and tetradecanoyl chloride.Example 61Tetradecanoic acid-2-[{3—[(3~dimethylamino-propyl)-methyl—amino]—propy1}—(2-tetradecanoyloxy-ethyl)—amino]-ethyl esterThe title compound is obtained as an oil in a 35 % yieldin an analogous manner to that described in example 1from 2-[{3-[(3-dimethylamino-propyl)—methyl—amino]-propyl}-(2-hydroxy-ethyl)-amino]-ethanol andtetradecanoyl chloride.Example 62Tetradecanoic acid-2—[{3—[(3-diethylamino-propy1)-methyl-amino]-propy1}—(2-tetradecanoyloxy-ethyl)-amino]-ethyl esterThe title compound is obtained as an oil in a 61 % yieldin an analogous manner to that described in example 1CA 02265765 l999-03- 10-45..from 2—[{3—[(3-diethylamino—propyl)-methyl-amino]-propyl}-(2-hydroxy-ethyl)-amino]-ethanol andtetradecanoyl chloride.Example 63N-<3-{4-[Bis-(2-tetradecanoyloxy—ethy1)-amino]-piperidin-1—yl}-propyl>-guanidineThe title compound is obtained as an oil in a 91 % yieldin an analogous manner to that described in example 44from the compound of example 22 and cyanamide.Example 64N-[3-(4-<[Bis-(2-tetradecanoyloxy-ethyl)-amino]-methyl>-piperidin-1—yl)-propyl]—guanidineThe title compound is obtained as an oil in an 82 %yield in an analogous manner to that described inexample 44 from the compound of example 24 andcyanamide.Example 65N-[2-(1-<3-[Bis-(2-tetradecanoyloxy—ethyl)-amino]-propyl>-piperidin-4-yl)~ethyl]-guanidine.The title compound is obtained as an oil in a 94 % yieldin an analogous manner to that described in example 44from the compound of example 40 and cyanamide.CA 02265765 l999-03- 10-46..Example 66Tetradecanoic acid-2-{(2—tetradecanoyloxy-ethyl)-[1-(2-amino—ethyl)-piperidin-4-yl]-amino}-ethyl esterThe title compound is obtained as an oil in a 38 % yieldin an analogous manner to that described in example 10from 2—{(2—hydroxy—ethyl)—[1-(2-amino-ethyl)-piperidin-4-yl]-amino}-ethanol and tetradecanoyl chloride.The 2-{(2—hydroxy-ethyl)—[1—(2—amino—ethyl)—piperidin-4-yl]-amino}—ethanol used as the starting material can beobtained as follows:a) A mixture of 8.5 g (45 mmol) of the 2-[(2-hydroxy—ethyl)-(piperidin—4—yl)-amino]-ethanol described inexample 3b), 150 ml dimethylformamide, 3.4 g (45 mmol)chloroacetonitrile, 14.5. g potassium carbonate and100 mg potassium iodide is stirred for 2 h at 60°C. Itis filtered and the filtrate is concentrated byevaporation in a vacuum. 10.5 g (quantitative) 2-{(2-hydroxy—ethyl)-[1-cyanomethyl-piperidin-4-yl]-amino}-ethanol remains as an oily crude product.b) 4.1 g (18 mmol) of the previously described compoundis hydrogenated in 175 ml methanolic ammonia over Raneynickel at 35°C and 100 bar hydrogen pressure. It isfiltered, concentrated by evaporation and afterchromatography on silica gel one obtains 2.3 g (55 % oftheory) 2{(2—hydroxy—ethyl)-[1—(2-amino-ethyl)—piperidin—4-yl]-amino}-ethanol as an oil.CA 02265765 l999-03- 10-47-Example 67Tetradecanoic acid—2—{(2-tetradécanoyloxy-ethyl)-[1-(2-amino-ethyl)-piperidin-4-yl-methyl]-amino}—ethyl esterThe title compound is obtained as an oil in a 29 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)—[1-(2-amino—ethyl)—piperidin-4-yl-methyl]-amino}~ethanol and tetradecanoyl chloride.The 2—{(2—hydroxy-ethyl)-[1-(2-amino-ethyl)-piperidin-4-y1—methyl]-amino}—ethanol used as the starting materialcan be obtained as follows:a) The intermediate product described in example 6b) isreacted with chloroacetonitrile analogously to themethod described in example 66a).b) The crude product obtained above is hydrogenated asin example 66b). The desired 2-{(2-hydroxy—ethyl)—[1-(2-amino-ethyl)-piperidin-4-yl—methyl]—amino}-ethanol isobtained in a yield of 47 % of theory.Example 68Tetradecanoic acid-2-{[3—(4-(aminomethyl)—piperidin-1-yl)-propyl]-(2-tetradecanoyloxy-ethyl)-amino}-ethylesterThe title compound is obtained as an oil in a 27 % yieldin an analogous manner to that described in example 10from 2—{[3—(4—(aminomethy1)—piperidin—1-yl)-propyl]—(2-hydroxy-ethyl)—amino}-ethanol and tetradecanoylCA 02265765 l999-03- 10-48-chloride.The 2-{[3-(4—aminomethyl-piperidin—1-yl)-propyl]-(2-hydroxy-ethyl)—amino}-ethanol used as the startingmaterial can be obtained analogously to the compounddescribed in example 28c from 4-aminomethyl—piperidine.Example 69Tetradecanoic acid-2-{(2—tetradecanoyloxy-ethyl)-[1-(3-dimethylamino-propyl)-piperidin-4-yl]-amino}—ethyl esterThe title compound is obtained as an oil in a 25 % yieldin an analogous manner to that described in example 10from 2-{(2-hydroxy—ethyl)—[1-(3-dimethylamino-propyl)-piperidin—4—yl]-amino}—ethanol and tetradecanoylchloride.The 2-{(2—hydroxy-ethyl)-[1-(3—dimethylamino—propyl)-piperidin-4-yl]-amino}—ethanol used as the startingmaterial is described in example 29.Example 70Tetradecanoic acid-2-{[3-(4-dimethylamino—piperidin-1-yl)-propyl]-(2-tetradecanoyloxy—ethyl)-amino}-ethylesterThe title compound is obtained as an oil in a 62 % yieldin an analogous manner to that described in example 10from 2—{[3-(4—dimethylamino-piperidin-1-yl)-propyl]-(2-hydroxy-ethyl)-amino}—ethano1 and tetradecanoylchloride.CA 02265765 l999-03- 10-49-The 2-{[3-(4-dimethylamino-piperidin-1-yl)-propyl]-(2-hydroxy-ethyl)-amino}-ethanol used as the startingmaterial is described in example 28.Example 71Tetradecanoic acid-2-{(2-tetradecanoy1oxy—ethyl)-[1-(4-amino—butyl)-piperidin-4-yl]—amino}—ethyl esterThe title compound is obtained as an oil in a 22 % yieldin an analogous manner to that described in example 10from 2-{(2—hydroxy—ethyl)—[1-(4—amino-butyl)-piperidin-4-yl]-amino}—ethanol and tetradecanoyl chloride.The 2-{(2-hydroxy—ethyl)-[1-(4-amino—butyl)-piperidin—4—yl]-amino}—ethanol used as the starting material can beobtained as follows:a) A mixture of 18.8 g (100 mmol) of the 2-[(2-hydroxy-ethyl)-(piperidin-4-yl)-amino]-ethanol described inexample 3b), 100 ml dimethyl-formamide, 11 ml (110 mmol)4-bromo-butyronitrile, 18.9 ml N-ethyldiisopropylamineand 100 mg 4-dimethyl-amino-pyridine is stirred for 8 hat 90°C and subsequently concentrated by evaporation ina vacuum. It is taken up in dichloromethane, filteredand the filtrate is concentrated. 15.0 g (59 % oftheory) 2-{(2-hydroxy-ethyl)—[1-(3-cyano~propyl)-piperidin-4-yl]-amino}-ethanol remains as an oil.b) 15.0 g (59 mmol) of the previously described compoundis hydrogenated in 300 ml methanolic ammonia over Raneynickel at 35°C and 100 bar hydrogen pressure. It isfiltered, concentrated by evaporation and 10.9 g (71 °o\CA 02265765 l999-03- 10-50-of theory) 2-{(2—hydroxy-ethyl)-[1-(4-amino-butyl)—piperidin—4—y1]-amino}-ethanol is obtained as an oil.Example 72Tetradecanoic acid-2-{(2—tetradecanoyloxy-ethyl)-[1-(4-dimethylamino-butyl)-piperidin-4-yl]-amino}—ethyl esterThe title compound is obtained as an oil in a 23 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)—[1-(4-dimethylamino-butyl)-piperidin—4-yl]—amino}—ethanol and tetradecanoylchloride.The 2—{(2-hydroxy-ethyl)—[1-(4-dimethylamino-butyl)-piperidin—4-yl]-amino}-ethanol used as the startingmaterial can be obtained as follows:4.7 ml formic acid and 5.6 ml saturated Formalinsolution are added dropwise at 0°C to 6.5 g (25 mmol) 2-{(2-hydroxy-ethyl)-[1-(4—amino-butyl)-piperidin-4—yl]-amino}-ethanol (example 71b) it is heated to 95 - 100°Cand stirred for a further 9 h. It is allowed to cool,admixed with 6.5 ml concentrated hydrochloric acid,refluxed for 3 h, made strongly alkaline with 10 Nsodium hydroxide solution and extracted withdichloromethane. After drying and concentrating theextract by evaporation, 6.0 g (84 % of theory) of thedesired compound remain as an oil.CA 02265765 l999-03- 10-51..Example 73Tetradecanoic acid—2-{(2-tetradécanoyloxy-ethyl)-[1-(4-amino-butyl)-piperidin-4-yl-methyl]-amino}—ethy1 esterThe title compound is obtained as an oil in a 33 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)-[1-(4-amino-butyl)-piperidin-4—y1-methyl]-amino}-ethanol and tetradecanoyl chloride.The 2-{(2—hydroxy-ethyl)—[1-(4-amino-butyl)—piperidin—4-yl-methyl]—amino}—ethano1 used as the starting materialcan be obtained as follows:The desired compound is obtained as an oil by reacting2-[(2—hydroxy—ethyl)-(piperidin-4-yl—methyl)-amino]-ethanol (example 6b) with 4-bromo-butyronitrileanalogously to example 71b) and subsequentlyhydrogenating analogously to example 71b).Example 74Tetradecanoic acid-2-{(2-tetradecanoyloxy-ethyl)-[1-(4-dimethylamino-butyl)~piperidin-4-yl-methyl]-amino}—ethylesterThe title compound is obtained as an oil in a 43 % yieldin an analogous manner to that described in example 10from 2-{(2-hydroxy-ethyl)-[1-(4-dimethylamino—butyl)-piperidin~4—yl—methyl]—amino}-ethanol and tetradecanoylchloride.The 2—{(2-hydroxy-ethyl)-[1-(4-dimethylamino—butyl)-CA 02265765 l999-03- 10-52..piperidin-4-yl—methyl]-amino}-ethanol used as thestarting material can be obtained (yield 69 % of theory)from the 2-{(2-hydroxy-ethyl)ell-(4-amino—butyl)—piperidin-4-yl-methyl]—amino}-ethanol described underexample 73 by the method described in example 72 byreaction with formic acid and Formalin solution.Example 75Tetradecanoic acid—2-{(2-tetradecanoyloxy-ethyl)-2—[[1-(4-amino-butyl)—piperidin-4—yl]-ethyl]—amino}-ethylesterThe title compound is obtained as an oil in a 17 % yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)-2-[[1-(4-amino-butyl)-piperidin—4-yl]-ethyl]-amino}-ethanol and tetradecanoylchloride.The 2—{(2—hydroxy—ethyl)—2—[[1—(4—amino-butyl)—piperidin—4—yl]—ethyl]~amino}-ethanol used as thestarting material can be obtained as follows:The desired compound is obtained as an oil by reactingthe 2-[(2-hydroxy-ethyl)—(2—piperidin—4-yl—ethyl)-amino]—ethanol described in example 12 with 4—bromo-butyronitrile analogously to example 71a) andsubsequently hydrogenating analogously to example 71b).CA 02265765 l999-03- 10-53-Example 76Tetradecanoic acid-2—{(2-tetradécanoyloxy-ethyl)-2-[[1-(4-dimethylamino-butyl)-piperidin-4-yl]-ethyl]-amino}-ethyl ester0The title compound is obtained as an oil in a 33 6 yieldin an analogous manner to that described in example 10from 2—{(2-hydroxy-ethyl)—2—[[1-(4-dimethylamino~butyl)-piperidin-4-yl]-ethyl]-amino}-ethanol and tetradecanoylchloride.The 2-{(2-hydroxy—ethyl)-2-[[1-(4—dimethylamino-butyl)—piperidin-4-yl]-ethyl]-amino}—ethanol used as thestarting material can be obtained (yield 72 % of theory)from the 2—{(2—hydroxy—ethyl)-2-[[1—(4—amino—butyl)—piperidin-4-yl]~ethyl]-amino}-ethanol described inexample 75 by reaction with formic acid and Formalinsolution according to the method described in example72.Example 77 Pharmacological testing1. Test principleThe testing of DOTAP or the compounds of the inventioncomprises transfection of the test cells, proteindetermination by means of the BCA method (Pierce) andcarrying out a CAT Elisa and these are described hereusing DOTAP (Boehringer Mannheim) as an example. Afterultrasonication the cationic lipid DOTAP formsunilamellar vesicles (liposomes) in aqueous solutionwhich spontaneously form stable complexes with the DNACA 02265765 l999-03- 10-54..(pCMV~CAT). These complexes adhere to the cell surface,fuse with the cell membrane and pCMV-CAT is releasedinto the cytoplasm. The transiently expressed CAT isdetected in the cell lysate.1.1 Test cellsHeLa, human cancer epithelial cell line from the cervix,ATCC CCL 2RPMI 1640, human cancer epithelial cell line from thenasal septum, ATCC CCL 30CALU 1, human cancer epithelial cell line from the lung,ECACC 93120818 or another suitable test cell line1.2 Test mediumThe composition of the test medium used for therespective cell line corresponds with that of therespective culture medium, only the content of FCS isreduced by 50 % (only 5 % FCS instead of 10 %).2. Determination procedure2.1 Feeding cells for the test— 2.0 ml cell suspension (1.5 x 105 cells/ml) areplaced into each well of a 6-well multiplate- incubate for 24 h at 37°C, 5 % CO22.2 Transfection mixtureThe DOTAP/DNA complexes are mixed in a sterile 96-wellround-bottom plate.Example;60 ul HBS (Hank's buffered saline) buffer is addedfirst, then 20 pl DOTAP is added. The DNA is diluted1:20 with HBS. Then 40 ul = 2 pg DNA is added to thetransfection mixtures and mixed thoroughly. The mixtureCA 02265765 l999-03- 10-55-is allowed to stand for 15 min at room temperature.Procedure for transfection: ’aspirate TM (transfection medium) from all wellsadd 2 ml TM/wellrapidly add the transfection mixturesthoroughly mix the content of the wells by gentleswirlingincubate the cells for 6 h at 37°C, 5 % CO2afterwards aspirate the DOTAP/DNA mixtureadd 2 ml TM/wellincubate the cells for 40-44 h, 37°C, 5 % CO22.3 Lysing the cellsaspirate the TM and wash the cells twice with 2 mlice-cold PBS/well, aspirate completelyadd 0.5 ml lysis buffer (from the CAT—Elisa kit,Boehringer Mannheim Co.) to the washed cells andallow to stand for 30 min at room temperatureafter 30 min transfer the lysates into Eppendorf cupsand centrifuge for 10 min at 13000 rpm using aBiofugeremove an aliquot from the supernatant for theprotein assay according to the BCA method. Theremainder is shock-frozen with liquid nitrogen andstored at —80°C until the CAT-Elisa is carried out.2.4 Protein determination of the lysates using the BCA(bicinchoninic acid) methodthe BSA (bovine serum albumin) solution contained inthe kit (Boehringer Mannheim Co.) is used to prepareprotein standards by dilution with lysis buffer10 pl of the protein standard solution, the blank(lysis buffer) and the unknown sample are pipettedCA 02265765 l999-03- 10..56_into a microtitre plate. 200 pl working solution isadded to each well and the plate is shaken for 30 minon a shaker. After incubating for 30 min at 37°C theyare measured at 550 nm in an ELISA reader. Thedetermination of the protein concentration is carriedout using an evaluation program.2.5 CAT—ElisaPrinciple:The CAT (chloroamphenicol acetyl transferase)-Elisaserves to quantitatively determine the CAT expression ineukaryotic cells after transfection with a plasmid thatcontains CAT as the reporter gene. The CAT-Elisa is asandwich enzyme immunoassay. Anti—CAT antibodies arebound adsorptively to the walls of the modules. In thefirst step CAT from cell extracts specifically binds tothe coated modules. In the second step the fixed CAT isbound by an ANTI—CAT antibody which is labelled withdigoxigenin (anti—CAT-DIG). Anti-CAT-DIG is detected inthe third step by a peroxidase-labelled antibody againstdigoxigenin (Anti-DIG—POD) and visualized in asubsequent substrate reaction.Procedure:The working steps of the CAT—Elisa are carried outaccording to the working instructions contained in thekit.The microtitre plate is measured at 405 nm and areference wavelength of 492 nm with an Elisa reader.CA 02265765 l999-03- 10-57-The pharmacological data are shown as an example in thefollowing table:relative transfection efficiency in a CAT-Assay (DOTAP=1)compound of exampleHeLa cellsCalu cells343.23.9603.23.4

Claims (18)

Claims

1. Pharmaceutical preparation containing at least one compound of formula I

, in which A denotes hydrogen, a group NR1R2, a group NR1(CH2)p NR3R4, a group (C=NH)NH2 or a pyridinyl residue, B and D are the same or different and each denotes a bond, a C1 to C6 alkylene residue or a group NR5-C2 to C6-alkylene, C denotes piperidinediyl or piperazinediyl, W and X are the same or different and each denotes a bond or a carbonyl group, Y and Z are the same or different and each denotes a saturated or unsaturated hydrocarbon residue with 7 to 24 carbon atoms, R1 to R5 are the same or different and each represents hydrogen or a C1 to C6 alkyl residue, m is an integer 0, 1 or 2 and if m equals 2 both residues C can be the same or different, n and o are the same or different and each denotes the integers 2, 3 or 4 and p denotes an integer from 2 to 6 as well as physiologically tolerated salts thereof, provided that hydrazine derivatives are not included and that m cannot be 0 if A denotes hydrogen or a group (C=NH)NH2 and B and D being the same or different represent a bond or an alkylene residue.

2. Compounds of formula I

, in which A denotes hydrogen, a group NR1R2, a group NR1(CH2)p NR3R4, a group (C=NH)NH2 or a pyridinyl residue, B and D are the same or different and each denotes a bond, a C1 to C6 alkylene residue or a group NR5-C2 to C6 alkylene, C denotes piperidinediyl or piperazinediyl, W and X are the same or different and each denotes abond or a carbonyl group, Y and Z are the same or different and each denotes a saturated or unsaturated hydrocarbon residue with 7 to 24 carbon atoms, R1 to R5 are the same or different and each represents hydrogen or a C1 to C6 alkyl residue, m is an integer 0, 1 or 2 and if m equals 2 both residues C can be the same or different, n and o are the same or different and each denotes the integers 2, 3 or 4 and p denotes an integer from 2 to 6 as well as physiologically tolerated salts thereof, provided that hydrazine derivatives are not included and that m cannot be 0 if A denotes hydrogen, a group NR1R2 or a group (C=NH)NH2 and B and D being the same or different represent a bond or an alkylene residue and in the case that m = 1, the group A-B cannot be hydrogen or C1-C4 alkyl and that they do not include the compounds __ octadecanoic acid-[(3-diethylamino-propyl)imino]-bis(methyl-2,1-ethanediyl)-ester-hydrochloride octadecanoic acid-[(3-dimethylamino-propyl)imino]di-3,1-propanediyl ester-dihydrochloride octadecanoic acid-[(3-dimethylamino-propyl)imino]di-3,1-propanediyl ester octadecanoic acid-[(3-dimethylamino-propyl)imino]di-2,1-ethanediyl ester-hydrochloride docosanoic acid-2-[(3-dimethylamino-propyl)-[2-[(1-oxododecyl)oxy]ethyl]amino]ethyl ester hexadecanoic acid-[[2-(ethylmethylamino)ethyl]-imino]di-2,1-ethanediyl ester octadecanoic acid-[[3-(dimethylamino)propyl]imino]di-2,1-ethanediyl ester octadecanoic acid-[[3-(dimethylamino)propyl]imino]di-2,1-ethanediyl ester-dihydrochloride octadecanoic acid-[[3-(dimethylamino)propyl]imino]di-2,1-ethanediyl ester N,N-Bis[3-(dodecyloxy)propyl]-1,2-ethanediamine octadecanoic acid-[[2-[(2-aminoethyl)amino]ethyl]-imino]-di-2,1-ethanediyl ester stearic acid-iminobis-(ethyleneiminoethylene)-ester-monoacetate.

3. Compounds as claimed in one of the claims 1-2, in which W and X denote CO.

4. Compounds as claimed in one of the claims 1-3, in which Y denotes C13H27 or C17H33.

5. Compounds as claimed in one of the claims 1-4, in which Z denotes C13H27 or C17H33.

6. Compounds of formula II
, in which A, B, C, D, n and o have the meanings as claimed in claim 1 and m = 1 or 2, provided that hydrazine derivatives are not included and in the case that m = 1, the group A-B cannot be hydrogen or C1-C3 alkyl.

7. Compounds as claimed in one of the claims 1-6, in which A denotes NH2 or N(CH3)2.

8. Compounds as claimed in one of the claims 1-7, in which B and D are the same or different and denote a bond, a C1 to C3-alkylene residue or, if m=0, an N(CH3)C3 to C4-alkylene residue.

9. Compounds as claimed in one of the claims 1-8, in which C denotes a piperidinediyl.

10. Compounds as claimed in one of the claims 1-9, in which m denotes 0 or 1.

11. Compounds as claimed in one of the claims 1-10, in which n and o denote 2.

12. Pharmaceutical preparations as claimed in one of the claims 1, 3-5, 7-11 which additionally contain a nucleic acid.

13. Pharmaceutical preparations as claimed in one of the claims 1, 3-5, 7-11, which additionally contain a therapeutic agent.

14. Use of compounds of formula I, , in which A, B, C, D, m, n and o have the meanings as claimed in one of the claims 1-11, provided that hydrazine derivatives are not included and that m cannot be 0 if A denotes hydrogen or a group (C=NH)NH2 and B and D being the same or different represent a bond or an alkylene residue, for the production of gene ferries and transfection vehicles containing a therapeutic agent.

15. Use of compounds as claimed in one of the claims 1-12 for the production of pharmaceutical compositions for gene therapy.

16. Use of compounds as claimed in one of the claims 1-13 for in vitro transfection.

17. Use of compounds as claimed in one of the claims 1-13 for the production of in vivo transfection reagents.

18. Use of compounds as claimed in one of the claims 1 to 13 for the production of drug combinations in cancer therapy, antiviral therapy, infection therapy and in diseases caused by dysregulation.