United States Patent (19) 11 4,206,215 Bailey 45 Jun. 3, 1980

54 ANTIMICROBIAL (56) References Cited BIS-4-(SUBSTITUTED-AMINO)-1- U.S. PATENT DOCUMENTS PYRIDINIUMALKANES 3,055,902 9/1962 Walker ...... 260/296 D 75 Inventor: Denis M. Bailey, East Greenbush, 3,786,058 1/1974 Edwards ...... 260/294.8 R N.Y. 73 Assignee: Sterling Drug Inc., New York, N.Y. OTHER PUBLICATIONS Walker et al., J. Org. Chem., vol. 26, pp. 2740-2747, 21 Appl. No.: 876,031 (1961). 22 Filed: Feb. 8, 1978 Austin, J. Pharm. Pharmacol., vol. 11, pp. 80-93, (1959). Primary Examiner-John D. Randolph Related U.S. Application Data Attorney, Agent, or Firm-Paul E. Dupont; B. Woodrow 63 Continuation-in-part of Ser. No. 721,315, Sep. 7, 1976, Wyatt abandoned, which is a continuation-in-part of Ser. No. 661,101, Feb. 25, 1976, abandoned. 57 ABSTRACT 30 Foreign Application Priority Data Bis-4-(R-amino)-1-pyridiniumalkanes are prepared by Feb. 11, 1977 (GB United Kingdom ...... 5784/77 reacting a 4-(R-amino)pyridine with an appropriate 51 Int. Cl...... C07D 213/02; A61R 31/44 disubstituted alkane. The compounds are useful as anti 52 U.S. C...... 424/263; 546/261; microbial agents. Certain species are also useful as den 546/264; 252/106 tal plaque-preventive agents. 58 Field of Search ...... 260/296 D; 252/106; 424/263; 546/261,264 45 Claims, No Drawings 4,206,215 1. 2 groups are linked through an alkylene group ANTIMICROBIAL bonded to the nitrogen atoms of the pyridine rings. BIS-(4-(SUBSTITUTED-AMINO)-1-PYRIDINIUM ALKANES SUMMARY OF THE INVENTION In a composition aspect, this invention relates to a CROSS-REFERENCE TO RELATED group of bis-(4-(R-amino)-1-pyridiniumlalkanes which APPLICATION are useful as antimicrobial agents. Certain of the com This application is a continuation-in-part of prior pounds are also useful as dental plaque-preventive copending application Ser. No. 721,315, filed Sept. 7 agents. 1976, now abandoned, in turn a continuation-in-part of 1 O In a further composition aspect the present invention application Ser. No. 661,101, filed Feb. 25, 1976 and provides an antimicrobial composition suitable for topi cal administration comprising an effective amount of a now abandoned. bis-(4-(R-amino)-1-pyridiniumlalkane and a compatible, pharmaceutically acceptable carrier. BACKGROUND OF THE INVENTION 15 1. Field of the Invention In another composition aspect, this invention relates The present invention relates to compositions of mat to a skin-cleansing composition comprising an antimi ter classified in the art of chemistry as bis-(4-(R-amino)- crobially effective amount of a bis-(4-(R-amino)-1- 1-pyridiniumalkanes, to a process for the preparation pyridinium-alkane, a compatible, pharmaceutically acceptable surfactant and a compatible, pharmaceuti thereof, to compositions containing these compounds 20 and to methods of using the same for controlling micro cally acceptable carrier. organisms and for preventing the formation of dental In yet another composition aspect, this invention plaque. provides an oral hygiene composition for the preven 2. Prior Art tion of dental plaque comprising an effective amount of Walker U.S. Pat. No. 3,055,902, issued Sept. 25, 1962 25 a dental plaque-preventive bis-(4-(R-amino)-1-pyridini discloses a group of bis-(4-amino-1-pyridinium)alkanes. umlalkane and a compatible, pharmaceutically accept No utility is disclosed for these compounds other than able carrier. as intermediates in the preparation of the corresponding In another composition aspect, the present invention bis-(4-amino-1-piperidino)alkanes which are stated to provides an antimicrobial composition suitable for ap have bacteriostatic and bactericidal effects. 30 plication to inanimate surfaces comprising an antimi G. N. Walker et al., J. Org. Chem. 26, 2740-7 (1961) crobially effective amount of a bis-(4-(R-amino)-1- disclose essentially the subject matter disclosed in the pyridiniumalkane in admixture with a compatible vehi above-noted Walker U.S. patent. cle. W. C. Austin et al., J. Pharm. Pharmacol. 11, 80-93 In a method aspect, the present invention provides a (1959) disclose 1,10-bis-(4-amino-1-pyridinium)decane 35 method for controlling microorganisms which com diiodide and 1,10-bis(4-acetamido-1-pyridinium)decane prises contacting said microorganisms with an antimi diiodide. It is stated that certain species among the crobially effective amount of a bis-(4-(R-amino)-1- large, diverse group of quaternary ammonium com pyridiniumalkane. pounds disclosed possess amebicidal, antibacterial, anti In a further method aspect, this invention is con filarial and trypanocidal activity, but no biological data 40 cerned with a method for treating teeth to prevent the are given for either of the above-named compounds. formation of dental plaque thereon which comprises Edwards U.S. Pat. No. 3,786,058, issued Jan. 15, 1974 contacting the teeth with an effective amount of a den discloses a group of bis(pyridinium quaternary salts) tal plaque-preventive bis-(4-(R-amino)-1-pyridinium al believed to be structurally more remote from the kane. claimed compounds than the above-noted Walker and 45 DETAILED DESCRIPTION INCLUSIVE OF THE Austin et al. compounds and having the formula PREFERRED EMBODIMENTS The invention sought to be patented resides in one of its composition aspects in the bis-(4-(R-amino)-1- pyridinium-alkanes having Formula I hereinbelow: SN n N +2 er eks: W 7 RNH / N-Yann \ NHR A * wherein inter alia: 55 R1 and R2 are alkyl of 6 to 14 carbon atoms; n A1 and A2 are a direct linkage; R R A3 is -NHCH2)NH-, m being 0 to 12; and wherein: X.X represents two monoanions or a dianion. It is Y is an alkkylene group containing from 4 to 18 car stated that the compounds possess antibacterial bon atoms and separating the two 4-(R-NH)-1- properties and that some are useful in dental hy pyridinyl groups by from 4 to 18 carbon atoms; giene for inhibiting the formation of dental plaque. R is an alkyl group containing from 6 to 18 carbon In the above compounds the pyridinium groups are atoms, a cycloalkyl group containing from 5 to 7 linked through an alkylene diamine chain bonded carbon atoms, benzyl, or phenyl substituted by to the carbon atoms of the pyridine rings and thus 65 methylenedioxy or one or two substituents selected constitute a class of compounds which are separate from the group consisting of halogen, lower-alkyl, and distinct from the instantly claimed bis lower-alkoxy, nitro, cyano and trifluoromethyl; (pyridinium)alkanes in which the pyridinium R1 is hydrogen or lower alkyl; 4,206,215 4. A is an anion; 1,10-Bis-(4-(2-ethylhexylamino)-1-pyridinium)-dec m is 1 or 3; ane dichloride, n is 1 or 2; 1,12-Bis-4-(hexylamino)-1-pyridiniumdodecane di x is 1, 2 or 3; and bromide and the corresponding dichloride, wherein (m)(2)=(n)(x). 1,12-Bis-(4-(heptylamino)-1-pyridiniumdodecane di These compounds are useful as antibacterial and anti bromide, fungal agents and are virucidally active against Herpes 1, 12-Bis-4-(octylamino)-1-pyridiniumdodecane di viruses. bromide, Included in the above group are the compounds hav 1,12-Bis-4-(nonylamino)-1-pyridiniumdodecane di ing Formula I wherein Y, A, R1, m, n and x have the 10 bromide, abovegiven meaning and wherein: 1,12-Bis-(4-(decylamino)-1-pyridiniumdodecane di (a) R is an alkyl group containing from 6 to 18 carbon bromide, atoms; 1,12-Bis-4-(dodecylamino)-1-pyridiniumdodecan (b) R is a cycloalkyl group containing from 5 to 7 dibromide, carbon atoms; 15 1,12-Bis-4-(2-ethylhexylamino)-1-pyridinium (c) R is phenyl substituted by methylenedioxy or one dodecane dibromide and the corresponding dichloride, or two substituents selected from the group consisting 1,14-Bis-4-(hexylamino)-1-pyridiniumtetradecane of halogen, lower-alkyl, lower-alkoxy, nitro, cyano and dibromide and the corresponding dichloride, trifluoromethyl; 1,14-Bis-4-(heptylamino)-1-pyridiniumtetradecane (d) R is benzyl. 20 dibromide and the corresponding dichloride, Certain of the compounds of Formula I are also use 1,14-Bis-4-(octylamino)-1-pyridinium tetradecane ful as dental plaque-preventive agents, for example: dibromide, 1,6-Bis-4-(octylamino)-1-pyridinium hexane dibro 1,10-Bis-(4-(octylamino)-1-pyridinium)decane bis(1- mide and the corresponding dichloride, oxo-2-pyridinethiolate), 1,6-Bis-4-(nonylamino)-1-pyridinium hexane dibro 25 mide and the corresponding dichloride, and, as particularly preferred species, 1,6-Bis-4-(decylamino)-1-pyridinium hexane dibro 1,9-Bis-(4-(heptylamino)-1-pyridiniumnonane dibro mide, mide, 1,6-Bis-4-(dodecylamino)-1-pyridinium hexane di 1,10-Bis-(4-(2-ethylhexylamino)-1-pyridinium-dec bromide, 30 ane dibromide, 1,7-Bis-(4-(heptylamino)-1-pyridinium heptane dibro 1,10-Bis-4-(octylamino)-1-pyridinium)decane dichlo mide, ride and the corresponding dibromide, 1,7-Bis-4-(octylamino)-1-pyridinium]heptane dibro 19-Bis-4-(octylamino)-1-pyridiniumnonane dibro mide, mide, 1,7-Bis-(4-(nonylamino)-1-pyridinium]heptane dibro 35 1,2-Bis-4-(heptylamino)-1-pyridiniumdodecane di mide, chloride, and 1,7-Bis-4-(decylamino)-1-pyridiniumheptane dibro 1,8-Bis-4-(octylamino)-1-pyridinium]octane dibro mide, mide. 1,7-Bis-4-(dodecylamino)-1-pyridinium]heptane di In a further composition aspect the invention sought bromide, to be patented resides in an antimicrobial composition 1,8-Bis-4-(heptylamino)-1-pyridinium]octane dibro suitable for topical administration which comprises an mide and the corresponding dichloride, effective amount of a compound having the Formula I 1,8-Bis-4-(octylamino)-1-pyridinium]octane dichlo hereinabove and a compatible, pharmaceutically ac ride, ceptable carrier. 1,8-Bis-4-(nonylamino)-1-pyridinium octane dibro 45 In another composition aspect, the invention sought mide, to be patented resides in a skin-cleansing composition 1,8-Bis-4-(decylamino)-1-pyridinium]octane dibro comprising an antimicrobially effective amount of a mide, compound having the Formula I hereinabove, a com 1,8-Bis-4-(dodecylamino)-1-pyridinium]octane di patible, pharmaceutically acceptable surfactant and a bromide, 50 compatible, pharmaceutically acceptable carrier. 1,8-Bis-4-(2-ethylhexylamino)-1-pyridinium-octane In another composition aspect the invention sought dibromide, to be patented resides in an oral hygiene composition 1,9-Bis-4-(heptylamino)-1-pyridiniumnonane di for the prevention of dental plaque comprising an effec chloride, tive amount of a compound selected from the group 1,9-Bis-4-(nonylamino)-1-pyridiniumnonane dibro 55 consisting of 1,9-bis-(4-(heptylamino)-1-pyridinium no mide, nane dibromide, 1,10-bis-4-(2-ethylhexylamino)-1- 1,9-Bis-4-(decylamino)-1-pyridiniumnonane dibro pyridinium decane dibromide, 1,10-bis-(4-(octylamino)- mide, 1-pyridinium decane dichloride, 1.9-bis-(4-(oc 1,9-Bis-(4-dodecylamino)-1-pyridinium nonane di tylamino)-1-pyridiniumnonane dibromide, 1,12-bis-(4- bromide, , , (heptylamino)-1-pyridiniumdodecane dichloride, 1,8- 1,10-Bis-4-(heptylamino)-1-pyridinium)decane di bis-(4-(octylamino)-1-pyridinium octane dibromide and bromide and the corresponding dichloride, 1,10-bis-4-(octylamino)-1-pyridinium decane dibro 1,10-Bis-4-(nonylamino)-1-pyridinium decane dibro mide and a compatible, pharmaceutically acceptable mide, carrier. - 1, 10-Bis-4-(decylamino)-1-pyridinium)decane dibro 65 In yet another composition aspect; the invention mide, sought to be patented resides in an antimicrobial com 1,10-Bis-4-(dodecylamino)-1-pyridinium decane di position suitable for application to inanimate surfaces bromide, ' ' ... comprising an effective amount of a compound having 4,206,215 5 6 the Formula hereinabove in admixture with a compati -continued ble vehicle. s'. I --2 In one of its method aspects the invention sought to be patented resides in the method for controlling micro RHN Naya-N NHR A * organisms which comprises contacting said microor 5 ganisms with an antimicrobially effective amount of a compound having Formula I hereinabove wherein Y, R R R1, A, m, n and X have the above given meanings, and Ib wherein: 10 In the formulas herein the alkylene group Y is a biva R is hydrogen, an alkyl group containing from 1 to 18 lent saturated aliphatic hydrocarbon radical containing carbon atoms, a cycloalkyl group containing from 5 to from 4 to 18, preferably from 8 to 12, carbon atoms 7 carbon atoms, benzyl, or phenyl substituted by meth arranged in a straight or in a branched chain and sepa ylenedioxy or one or two substituents selected from the rating the two 4-(R-NH)-1-pyridinyl groups by from 4 group consisting of halogen, lower-alkyl, lower-alkoxy, 15 to 18, preferably from 8 to 12, carbon atoms, for exam nitro, cyano and trifluoromethyl. ple: A preferred embodiment of the above-described 1,4-butylene, 1,5-pentylene, 1,6-hexylene, 1,7-hepty method resides in the use therein as antimicrobial agent, lene, 1,8-octylene, 1,9-nonylene, 1,10-decylene, 1,11 a compound having Formula I wherein R is an alkyl undecylene, 1,12-dodecylene, 1,13-tridecylene, 1,14-tet group containing from 1 to 18 carbon atoms, a cycloal 20 radecylene, 1,15-pentadecylene, 1,16-hexadecylene, kyl group containing from 5 to 7 carbon atoms, benzyl, 1,17-heptadecylene, 1, 18-octadecylene, 1-methyl-1,4- or phenyl substituted by methylenedioxy or one or two butylene, 3-methyl-1,5-pentylene, 2-ethyl-1,4-butylene, substituents selected from the group consisting of halo 3-methyl-1,6-hexylene, 2,4-dimethyl-1,5-pentylene, 1 gen, lower-alkyl, lower-alkoxy, nitro, cyano and trifluo methyl-1,7-heptylene, 3-ethyl-1,6-hexylene, 3-propyl romethyl. 25 1,5-pentylene, 4,4-dimethyl-1,7-heptylene, 2,6-dimeth Also included within the ambit of the above method yl-1,7-heptylene, 2,4,4-trimethyl-1,6-hexylene, 2,7- is the use of compounds wherein R is hydrogen. Several dimethyl-1,8octylene, 1-methyl-1,10-decylene, 5-ethyl of these compounds are known in the prior art e.g. the 1,9-nonylene, 3,3,6,6-tetramethyl-1,8-octylene, 3,8- Walker and Austin et al. references cited hereinabove. 30 dimethyl-1,10-decylene, 3-methyl-1,11-undecylene, 6 However, these references disclose no utility for these methyl-1,12-dodecylene, 2-methyl-1, 13-tridecylene, compounds except as intermediates. Accordingly, the 4,9-dimethyl-1,12-dodecylene, 4-methyl-1,14-tetradecy discovery of antimicrobial activity therefor, albeit of lene, 2,13-dimethyl-1,14-tetradecylene, 1,4-dipropyl substantially lower potency and narrower spectrum 1,4-butylene, 3-(3-pentyl)-1,5-pentylene, 2-(4,8-dime 35 thylnonyl)-1,4-butylene, 1-heptyl-1,5-pentylene, and the relative to the novel compounds, is considered within like. the scope of the present invention. It will be appreciated that when Y contains 4 carbon In another of its method aspects the invention sought atoms the latter must, of course, be arranged in a to be patented resides in the method of treating teeth for straight chain and similarly when Y contains 18 carbon preventing the formation of dental plaque thereon atoms and separates the two 4-(R-NH)-1-pyridinyl which comprises contacting the teeth with an effective groups by 18 carbon atoms said carbon atoms must also amount of a compound selected from the group consist be arranged in a straight chain. In all other instances Y ing of 1.9-bis-(4-heptylamino)-1-pyridinium nonane di can be either straight or branched. bromide, 1,10-bis-(4-(2-ethylhexylamino)-1-pyridini In the formulas herein when R is a straight or umldecane dibromide, 1,10-bis-4-(octylamino)-1- 45 branched-chain alkyl group containing from 6 to 18, pyridinium decane dichloride, 1,9-bis-4-(octylamino)-1- there are included, for example: pyridinium nonane dibromide, 1,12-bis-(4-(hep n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-unde tylamino)-1-pyridiniumdodecane dichloride, 1,8-bis(4- cyl, n-dodecyl, n-tridecyl, n-tetradecyl, n-pentadecyl, (octylamino)-1-pyridinium]octane dibromide and 1,10 n-hexadecyl, n-heptadecyl, n-octadecyl, 1-methylpen bis-(4-(octylamino)-1-pyridinium)decane dibromide. 50 tyl, 2,2-dimethylbutyl, 2-methylhexyl, 1,4-dimethylpen Although the compounds of the present invention are tyl, 2-ethylpenty, 3-ethylpentyl, 2-methylheptyl, 1 conveniently represented by Formula I hereinabove, it ethylhexyl, 2-ethylhexyl, 2-propylpentyl, 2-methyl-3- will of course, be appreciated that electron-delocalized ethylpentyl, 3-ethylheptyl, 1,3,5-trimethylhexyl, 1,5- or resonance forms of Formula I (i.e. differing only in dimethyl-4-ethylhexyl, 2-propylhepty, 5-methyl-2- 55 butylhexyl, 2-propylnonyl, 2-butyloctyl, 1,1-dime the position of electrons) such as Formulas Ia and Ib thylundecyl, 2-pentylnonyl, 1,2-dimethyltetradecyl, hereinbelow are equivalent representations of the in 1,1-dimethylpentadecyl and the like. Straight or stant compounds. The actual structure of the com branched-chain alkyl groups containing from 7 to 9 pounds in a resonance hybrid of Formulas I, Ia and Ib. carbon atoms are preferred. 60 When R in the formulas herein is a cycloalkyl group containing from 5 to 7 carbon atoms, there are included 2 cyclopentyl, cyclohexyl, cycloheptyl and the like. / W When R in the formulas herein is phenyl substituted RNH N-Y-N NHR A* by methylenedioxy or one or two substituents selected 65 from the group consisting of halogen, lower-alkyl, low er-alkoxy, nitro, cyano and trifluoromethyl, there are R Rt included p-chlorophenyl, o-chlorophenyl, m a chlorophenyl, p-bromophenyl, m-fluorophenyl, p 4,206,215 7 8 iodophenyl, 2,4-dichlorophenyl, 2,4-difluorophenyl, The reaction is conveniently carried out by reacting 2,5-dibromophenyl, 3,5-dichlorophenyhl, 3-chloro-4- two moles of a 4-(R-amino)pyridine (Formula II) with fluorophenyl, 3,4-methylenedioxyphenyl, p-ethylphe one mole of an appropriately disubstituted alkane (For nyl, p-methoxyphenyl, m-nitrophenyl, o-cyanophenyl, mula III) in an inert solvent such as a lower alkanol, m-(trifluoromethyl)phenyl, 2-methoxy-5-methylphenyl, acetonitrile, N,N-dimethylformamide, N,N-dime and the like. thylacetamide, benzene, toluene, xylene and the like, at In the formulas herein R can also be benzyl. If de a temperature of from about 80 C. to 150° C. for a sired, the phenyl moiety of the benzyl group can be period of from 2 to 24 hours. Usually the reactants are substituted with from one to two substituents such as for heated at 80 C. to 100° C. in acetonitrile or N,N-dime example, halogen, hydroxy, lower-alkyl, lower-alkoxy, O thylformamide or a mixture of these solvents for about nitro, cyano, trifluoromethyl, and the like. . 15 to 20 hours. In Formula I, R is the same in both occurrences. Riis Alternatively the reaction may be carried out in the also the same in both its occurrences in Formula I. absence of a solvent by heating stoichiometric quantities When A in the formulas herein is an anion there are of the reactants at 120° C.-150° C. for about 2 to 5 included anions of both inorganic and organic acids for 15 hours. example: bromide, chloride, fluoride, iodide, sulfate, phosphate, monofluorophosphate, nitrate, sulfamate, The resulting bis-(4-(R-amino)-1-pyridiniumalkanes methanesulfonate, ethanesulfonate, benzenesulfonate, (Formula I) are isolated according to conventional p-toluenesulfonate, naphthalenesulfonate, naph methods, for example by filtration, if the product is thalenedisulfonate, cyclohexylsulfamate, acetate, tri 20 insoluble in the reaction medium, or by dilution of the fluoroacetate, malate, fumarate, succinate, tartrate, tar reaction mixture with a non-polar solvent such as ether, tronate, oxalate, citrate, lactate, gluconate, ascorbate, benzene or hexane in order to precipitate the productor lactate, phthalate, salicylate, benzoate, picrate, me by evaporation of the reaction medium to leave the thanephosphate, ethane-1-hydroxy-1,1-diphosphonate, product as a residue. The isolated crude product can be arsenite, arsenate, thiosulfate, perchlorate, sarcosinate, 25 purified by crystallization from a suitable solvent in the N-lauroylsarcosinate, nitrilotriacetate, 2-hydroxyethyl presence of an adsorbent, e.g. charcoal or diatomaceous nitrilodiacetate, zinc phenolsulfonate, 1-oxo-2-pyridine earth. thiolate The bis-(4-(R-amino)-1-pyridiniumalkanes produced in accordance with the above-described method will, of 30 course, contain an anion (A in Formula I) which corre 21 sponds to the leaving group of the reactant disubstituted alkane (Z in Formula III). However, the anionic moiety of these compounds can is . be varied, if desired, according to conventional ion v 35 exchange methods, for example by passing a solution of O a pyridinium compound in a suitable solvent, e.g. meth anol, ethanol or water through a bed of synthetic ion tetrafluoroborate, hexachloroplatinate, hexa exchange resin containing the desired anion. The sol fluoroaluminate, tetrachloroaluminate, hexafluorostan vent is evaporated and the resulting pyridinium com nate, hexachlorostannate, hexafluorosilicate, fluorozir pound containing the desired anion is purified by re conate and the like. Bromide and chloride are preferred. crystallization from a suitable solvent. The term "halogen' as used herein is intended to Alternatively, a pyridinium compound can be reacted include fluorine, chlorine, bromine and iodine. with a soluble salt containing the desired anion in asso In the terms lower-alkyl and lower-alkoxy, "lower' ciation with a counter-cation which combines with the denotes an alkyl moiety containing from 1 to 4 carbon 45 anion of the pyridinium compound to produce an insol atoms such as methyl ethyl, propyl, isopropyl, butyl, uble precipitate. The latter is separated leaving a solu isobutyl and tert-butyl. tion of the pyridinium compound containing the desired The compounds of Formula I hereinabove are ob anion. For example a bis(4-(R-amino)-1-pyridiniumlal tained by reacting a 4-(R-amino)pyridine having For kane dihalide is reacted with the silver salt of an organic 50 or inorganic acid. The precipitated silver halide is re mula II hereinbelow: moved leaving a solution of the pyridinium compound containing the desired organic or inorganic anion. II The above type of metathetical reaction can also be employed to prepare insoluble pyridinium compounds. 55 Thus a soluble bis-(4-(R-amino)-1-pyridiniumlalkane is reacted with a soluble salt containing the desired anion which combines with the pyridinium cation to afford the desired product as an insoluble precipitate. These with a disubstituted alkane having Formula III herein insoluble pyridinium salts are useful for purposes of below: isolation and purification and when suitably composited as emulsions, creams, pastes, lotions, jellies or powders Z-Y-Z (III) also serve as depot preparations providing slow, sus tained release of the antimicrobial pyridinium com wherein in Formulas II and III R, R1 and Y have the pound. Moreover, insoluble salts derived from certain previously given meanings and Z is selected from the 65 anions such as those described in U.S. Pat. No. group consisting of chloro, bromo, iodo, methanesul 3,937,807, issued Feb. 10, 1976 are effective in reducing fonyloxy, ethanesulfonyloxy, benzenesulfonyloxy and the tooth staining potential of dental plaque preventive p-toluenesulfonyloxy. agents. 4,206,215 10 Thus, should the anionic portion of a given com tive bacteria, several species of fungi and Herpes vi pound accord to that species characteristics such as ruses. The compounds are therefore indicated for use as solubility, stability, molecular weight, physical appear antimicrobial or antiseptic agents which can be applied ance toxicity or the like, which render that form of the topically to effect the degerming of human skin and compound unsuitable for a desired purpose, it can other tissues and to sanitize and disinfect inanimate readily be converted to another, more suitable form. surfaces. Thus, the compounds can be used in topical For use on the skin or other tissues or in the oral cavity, antiseptic solutions for the treatment of wounds, in pharmaceutically acceptable anions such as, fluoride, antibacterial cleansing agents such as surgical hand chloride, bromide, iodide, methanesulfonate, and the scrubs, patient pre-operative skin preparations, soaps, like, are of course employed. 10 and shampoos, or in household and industrial cleaners, The 4-(R-amino)pyridines (Formula II) which are disinfectants and protective coverings such as paints, used as starting materials generally are known or, if varnishes and waxes. The compounds are adapted for specifically new, are prepared according to the pro the above indicated utilities by combining them with ceudres described for preparation of the known com conventional diluents or carriers, compatible cationic, pounds. . 15 anionic or non-ionic surfactants, buffering agents, per Conveniently, the 4-(R-amino)pyridines are prepared fumes and coloring agents, and are applied to a surface by reacting 4-chloro or 4-bromopyridine or N-(4- to be disinfected by conventional methods, such as pyridyl)-pyridinium chloride hydrochloride with an scrubbing, spraying, swabbing, immersion and the like. appropriately substituted amine, The reaction is usually For use as skin-cleansing agents the bis-(4-(R-amino)- carried out by heating the reactants in the absence of a 20 1-pyridiniumalkanes can be prepared as liquids, or, if solvent at 150' C.-225' C. for about 1.5 to 3 hours. The desired, the liquid formulations can be thickened by product is isolated in a conventional manner, for exam certain additives into a gel or paste or molded into a bar pleby extraction from alkaline aqueous medium into an according to methods known in the art. For example, organic solvent such as ether, methylene chloride or the compounds can be formulated with any compatible, chloroform, evaporation of the organic solvent and 25 pharmaceutically acceptable surfactant preferably a crystallization of the residue from an appropriate sol non-ionic surfactant such as the polyoxyethylene poly Vent, oxypropylene copolymers described in U.S. Pat. No. Alternatively, the 4-(R-amino)pyridines are obtained 3,855,140, amine oxides such as stearyl dimethyl amine by catalytic hydrogenation of a mixture containing oxide described in U.S. Pat. No. 3,296,145 and the like 4-amino-pyridine and a carbonyl compound containing 30 or with mixtures of these. The formulations may addi the appropriate carbon content. The reaction is usually tionally contain pharmaceutically acceptable diluents carried out at a temperature of 50'-70' C. in a suitable such as water, lower alkanols and the like, acids, bases, solvent, for example ethanol, under a hydrogen pres or buffering agents so as to maintain a pH of 5.0 to 7.5, sure of 20-60 psi, in the presence of a palladium hydro and, optionally, perfumes and coloring agents. The genation catalyst. A hydrogenation time of 4-10 hours 35 bis-(4-(R-amino)-1-pyridiniumalkane component of is generally satisfactory. The use of a large excess of the such formulations is generally present in a concentra carbonyl compound, i.e. 200% or greater advanta tion of approximately 0.5 to 2.0 percent by weight, geously results in high yields of pure product in a reac preferably 1.0 percent by weight. tion time of 5 hours or less, Following removal of the When prepared as a tincture the bis-(4-(R-amino)1- catalyst the product is isolated by evaporation of the pyridiniumalkanes may be formulated with water, a solvent and either distilling the residue or crystallizing lower alkanone, e.g. acetone, and a lower alkanol such the latter from a suitable solvent. as ethanol. If desired the tincture may be tinted with a Reaction of an aldehyde having the appropriate car coloring agent. The active ingredient is generally pres bon content with 4-aminopyridine in the presence of entin a concentration of about 0.05 to 1.0 percent (w/v) formic acid at elevated temperatures also provides the 45 preferably 0.1 percent (w/v). 4-(R-amino)pyridines. Alternatively, the compounds can be formulated in The 4-(R-amino)pyridines can also be obtained by suitable pharmaceutical vehicles for treating bacterial acylation of 4-aminopyridine with an acyl halide having and fungal infections for example as lotions, ointments, the appropriate carbon content followed by reduction or creams by incorporating them in conventional lotion, of the resulting amide. The acylation is carried out 50 ointment or cream bases, such as alkyl polyether alco according to art-recognized methods, for example by hols, cetyl alcohol, stearyl alcohol and the like, or as reacting 4-aminopyridine with an acyl halide in an inert powders by incorporating them in conventional pow solvent such as methylene dichloride or chloroform in der bases such as starch, talc and the like, or as jellies, by the presence of an acid acceptor such as triethylamine. incorporating them in conventional jelly bases such as The amide so-produced is then reduced with a complex 55 glycerol and tragacanth. They can also be formulated metal hydride such as lithium aluminum hydride in a for use as aerosol sprays or foams. suitable solvent such as tetrahydrofuran, ether or diox When used for sanitizing and disinfecting inanimate ane and the amine product isolated in accordance with surfaces the compounds can be formulated with known known procedures. detergents and builders such as trisodium phosphate, The disubstituted alkanes having Formula III also 60 borax and the like. The bis-(4-(R-amino)-1-pyridinium used as starting materials generally are known com alkane component of such formulations is generally pounds, or if specifically new can be prepared accord present in a concentration up to about 10 percent by ing to the procedures used for preparing the known weight. compounds. . . . . As described in detail hereinbelow certain of the As described more fully hereinbelow the compounds 65 compounds of Formula I are effective in preventing the having Formula I exhibited in vitro antimicrobial activ formation of dental plaque. When intended for such use ity against several species of microorganisms among the compounds can conveniently be applied to the teeth which are included both gram positive and gram nega in the form of a mouthwash or a dentifrice. The com 4,206,215 11 12 pounds can be composited with conventional ingredi C. 4-(Heptylamino)pyridine was also prepared by ents used in mouthwash or dentifrice formulations, for hydrogenating a mixture containing 4-aminopyridine example water, alcohol, glycerin, buffers, thickeners, and heptaldehyde according to the procedure described flavoring and coloring agents. The bis-(4-(R-amino)-1- in Example 9B hereinbelow. pyridiniumalkane is usually present in such formula D. To a stirred warm solution containing 100 g. tions in a concentration of about 0.005 to 0.05 percent (0.052 mole) of 4-(heptylamino)pyridine in 40 ml. of by weight preferably about 0.01 percent by weight. acetonitrile was added dropwise a solution containing It will of course be appreciated that the vehicles, 9.3 g (0.026 mole) of 1,14-dibromotetradecane in 230 diluents, carriers and additives contained in the above ml. of acetonitrile and the resulting mixture was heated described formulations are compatible with the active O 20 hours under reflux. The product which precipitated ingredient, i.e. the antimicrobial effectiveness of the upon cooling the reaction mixture to room temperature bis-(4-(R-amino)-1-pyridiniumalkanes is not vitiated by was collected by filtration, washed with cold acetoni effects ascribable to the nature of the vehicle, diluent, trile and dried affording 13.9 g of 1,14-bis-(4-(hep carrier or other additive. tylamino)-1-pyridinium-tetradecane dibromide, m.p. The molecular structures of the compounds of the 15 88°-90° C. invention were assigned on the basis of study of their infrared and NMR spectra, and confirmed by the corre EXAMPLE 2 spondence between calculated and found values for A solution containing 5.0 g of 1,14-bis-(4-(hep elemental analyses for the elements. tylamino)-1-pyridiniumtetradecane dibromide in 5.0 The invention is illustrated by the following examples 20 ml. of methanol was added to the top of a 3-inch diame without, however, being limited thereto. ter column containing 500 ml. of synthetic anion ex change resin in the chloride form (sold by Rohm and EXAMPLE 1. Haas under the tradename Amberlite IRA 400) packed A. A mixture containing 130 g. (0.67 mole) of 4 in methanol and eluted with five 125-ml. portions of bromopyridine hydrochloride and 152 g (1.0 mole) of 25 methanol. The combined eluate was evaporated to dry n-heptylamine hydrochloride was heated in an oil bath. ness under reduced pressure and the residual oil was When the bath temperature reached 180°-185° C. the redissolved in ethanol, treated with decolorizing carbon reaction mixture began to melt and stirring was begun. and evaporated to dryness. The residual solid was tritu At 190-195 C. melting was complete and the liquid rated with ether containing a few drops of acetonitrile, mixture was stirred and heated at 210-220 C. for 2.5 30 collected by filtration and dried over phosphorus pen hours. The reaction mixture was then cooled to room toxide under vacuum to give 3.94 g. of 1,14-bis-(4-(hep temperature and the resulting solid was dissolved in tylamino)-1-pyridinium tetradecane dichloride, m.p. water, made alkaline with 35% aqueous sodium hydrox 113-116 C. ide and the product extracted with chloroform. The EXAMPLE 3 chloroform extracts were dried over anhydrous sodium 35 sulfate and evaporated to dryness under reduced pres A stirred suspension containing 11.54 g (0.06 mole) sure. The resulting viscous oil was diluted with a small of 4-(heptylamino)pyridine in 75 ml. of acetonitrile was amount of n-hexane and cooled to give a solid which heated under reflux until a clear homogeneous solution was collected by filtration and air-dried to afford 86.6 g. was formed. To the clear solution was then added drop of 4-(heptylamino)pyridine, m.p. 49-51 C. wise a warm solution containing 9.84 g (0.03 mole) of B. Alternatively, 4-(heptylamino)pyridine was pre 1,12-dibromododecane in 75 ml. of acetonitrile. When pared as follows: A mixture containing 229 g. (1.0 mole) the addition was complete, heating under reflux was of N-(4-pyridyl)pyridinium chloride hydrochloride and continued 18 hours. After cooling to room temperature, 228 g. (1.5 moles) of n-heptylamine hydrochloride was the reaction mixture was evaporated to dryness under heated 2 hours with stirring in an oil bath at a bath 45 reduced pressure. The residual solid was slurried in temperature of 215 C. The reaction mixture was cooled ether, collected by filtration and dried 48 hours under to 80 C., diluted with ice-water, made alkaline with vacuum at 60° C. to afford 21.1 g of 1,12-bis-(4-(hep 35% aqueous sodium hydroxide and extracted succes tylamino)-1-pyridiniumdodecane dibromide, m.p. sively with ether and chloroform. The organic extracts 101-103 C. were combined and evaporated to dryness under re 50 duced pressure. The residual viscous oil was dissolved EXAMPLE 4 in ether and the ethereal solution was washed with A. A solution containing 6.6 g. of 1,12-bis-(4-(hep water. The aqueous wash was back-extracted with tylamino)-1-pyridinium)dodecane dibromide in 25 ml. chloroform and the chloroform extracts were combined of methanol was added to the top of a 3-inch diameter with the ethereal solution. The combined organic solu 55 column packed with 1 liter of synthetic anion exchange tions were dried over anhydrous sodium sulfate and resin in the chloride form (sold by Rohm and Haas evaporated to dryness under reduced pressure. Cooling under the tradename Amberlite IRA 400) in methanol the residual oil to -78° C. effected partial solidification. and eluted slowly with 100-ml. portions of methanol The semi-solid was diluted with a small amount of ether until 700 ml. of eluate had been collected. The com and filtered. The solid so-obtained was dissolved in a bined eluate was evaporated to dryness under reduced mixture of acetonitrile and chloroform, the resulting pressure below 25 C. The residual gum was triturated solution treated with decolorizing carbon, filtered, and repeatedly with a 6 to 1 mixture of ether and acetonitrile the filtrate evaporated to dryness under reduced pres and dried under vacuum to afford 5.0 g of 1,12-bis-(4- sure. The resulting semi-solid was diluted with a small (heptylamino)-1-pyridiniumdodecane dichloride, m.p. amount of ether and cooled. The solid thus-produced 65 109-112 C. was collected by filtration and washed with a small B. Alternatively, a mixture containing 76.8 g. (0.4 volume of cold ether to give, after drying, 84.6 g. of mole) of 4-(heptylamino)pyridine and 47.8 g. (0.2 mole) 4-(heptylamino)pyridine, m.p. 50-52 C. of 1,12-dichlorododecane was heated 4 hours at 4,206,215 13 14 125-130' C. After slight cooling 300 ml. of acetonitrile resulting oil crystallized from acetonitrile. The product was added and the resulting mixture was heated at was collected by filtration and dried 48 hours at 70 steam bath temperature to effect complete solution then C./0.1 mm. to give 13.4 g of 1,14-bis-(4-(hexylamino)-1- stored in a refrigerator overnight. The precipitated pyridinium tetradecane dibromide, m.p. 91-93 C. product was collected by filtration, washed with cold acetonitrile and ether, and the hygroscopic product EXAMPLE 6 immediately dried under vacuum to give 112 g of 1,12 Following a procedure similar to that described in bis-(4-(heptylamino)-1-pyridiniumdodecane dichloride, Example 2 but employing 5.0 g. of 1,14-bis-4-(hex m.p. 112-115° C. ylamino)-1-pyridinium tetradecane dibromide there O was obtained 4.33 g of the corresponding dichloride, EXAMPLE 5 m.p. 94-95 C. A. A mixture containing 100.0 g (0.51 mole) of 4 bromopyridine hydrochloride and 110 g (0.8 mole) of EXAMPLE 7 n-hexylamine hydrochloride was heated in an oil bath. Following a procedure similar to that described in When the bath temperature reached 175°-180° C. the 15 Example 5C but employing 10.7 g (0.06 mole) of 4 reaction mixture began to melt and stirring was begun. (hexylamino)pyridine and 9.85 g (0.03 mole) of 1,12 The temperature of the bath was then raised to 227 C. dibromododecane there was obtained 17.6 g. of 1,12-bis and the stirring continued 3.5 hours. After cooling to 4-(hexylamino)-1-pyridiniumdodecane dibromide, room temperature the reaction mixture was dissolved in m.p. 122-124 C. hot water, the resulting solution cooled with ice, made 20 alkaline with dilute aqueous sodium hydroxide and EXAMPLE 8 extracted with chloroform. The chloroform extracts Following a procedure similar to that described in were dried over anhydrous sodium sulfate and evapo Example 2 but employing 5.0 g of 1,12-bis-(4-(hex rated to dryness under reduced pressure. The residue ylamino)-1-pyridiniumdodecane dibromide there was was triturated with ether and cooled. The resulting 25 obtained 3.69 g. of the corresponding dichloride, m.p. solid was collected by filtration and washed with cold 86-88 C. ether. Evaporation of the filtrate afforded a second crop of solid. The crops were combined, dissolved in chloro EXAMPLE 9 form, treated with decolorizing carbon and filtered. A. A mixture containing 183.3 g (0.8 mole) of N-(4- The filtrate was evaporated under reduced pressure, 30 pyridyl)pyridinium chloride hydrochloride and 162 g. and the residue was triturated with cold ether. The (0.98 mole) of n-octylamine hydrochloride was heated product thus-obtained was collected by filtration, in an oil bath to a bath temperature of 225-230 C. washed with cold ether and dried to give 63.6 g of (internal temperature of 188 C.) and the resulting liquid 4-(hexylamino)pyridine, m.p. 66'-68 C. Evaporation of was stirred at that temperature for 2.5 hours. The reac the filtrate afforded an additional 7.0 g, m.p. 65-67 C. 35 tion mixture was then cooled to 70° C., diluted with 1 B. Alternatively, 4-(hexylamino)pyridine was pre liter of ice and water, made alkaline with 35% aqueous pared as follows. A mixture containing 229 g. (1 mole) sodium hydroxide and extracted with chloroform. The of N-(4-pyridyl)pyridinium chloride hydrochloride and chloroform extracts were dried over anhydrous sodium 207 g. (1.5 moles) of n-hexylamine hydrochloride was sulfate, treated with decolorizing carbon and evapo stirred and heated 1.75 hours at 175-185 C. The reac rated to dryness under vacuum. The resulting oil was tion was cooled and diluted with 750 ml. of ice-water. cooled to -78 C. The semi-solid which formed was The resulting solution was made alkaline with 35% triturated with ether and the solid thus-obtained col aqueous sodium hydroxide and after further dilution lected by filtration, washed with cold ether and dried. with 1 1. of water was extracted with ether followed by The filtrate afforded a second crop of 10 g. The crops dichloromethane. The organic extracts were combined, 45 were combined, dissolved in chloroform and, following dried over anhydrous sodium sulfate and evaporated to treatment with decolorizing carbon and filtration (re dryness under reduced pressure. The residue was crys peated 3 times), the chloroform solution was evapo tallized from ether, redissolved in chloroform, the re rated to dryness under reduced pressure. The resulting sulting solution treated with decolorizing carbon and solid was triturated with ether, cooled, collected by filtered. The filtrate was evaporated to dryness under 50 filtration and washed with cold ether-hexane to give reduced pressure and the residue was triturated with a 63.3 g or nearly colorless 4-(octylamino)pyridine, m.p. mixture of ether and acetonitrile. The solid thus 62-63 C. obtained was again dissolved in chloroform and the B. Alternatively 4-(octylamino)pyridine was pre resulting solution treated with decolorizing carbon and pared as follows: A mixture containing 94 g. (1 mole) of filtered. The filtrate was evaporated to dryness under 55 4-aminopyridine, 384 g. (3 moles) of octaldehyde, 7 g. of reduced pressure and the residue was triturated with 10% palladium-on-carbon hydrogenation catalyst and cold ether to give 71.0 g of 4-(hexylamino)pyridine, sufficient absolute ethanol to give a total volume of 1.2 m.p. 68-70° C. l. was hydrogenated 4.5 hours at 70°-90° C. under an C. To a stirred warm solution containing 10.7 g (0.06 initial hydrogen pressure of 45 psi. After cooling the mole) of 4-(hexylamino)pyridine in 50 ml. of acetonitrile mixture the hydrogenation catalyst was removed by was added dropwise a solution containing 10.7 g (0.03 filtration and the filtrate was evaporated to dryness mole) of 1,14-dibromotetradecane in 250 ml. of acetoni under reduced pressure. The residual oil crystallized on trile and the resulting mixture heated 22 hours under standing and the solid product was triturated with hex reflux. The reaction was then evaporated to dryness ane, collected by filtration, washed with fresh hexane under reduced pressure. The residual solid was dis 65 and dried at 40 C. under vacuum to give 182 g of solved in acetonitrile and the resulting solution treated 4-(octylamino)pyridine, m.p. 70-73 C. with decolorizing carbon and filtered. The filtrate was C. In another preparation a finely ground mixture evaporated to dryness under reduced pressure and the containing 500 g. (3.33 moles) of 4-chloropyridine hy 4,206,215 15 16 drochloride and 55 g (3.33 moles) of octylamine hydro chloride was gradually heated to an internal tempera EXAMPLE 12 ture of 180 C, in a nitrogen atmosphere and vigorous Following a procedure similar to that described in stirring was begun as soon as it became possible. The Example 2 but employing 5.0 g of 1,8-bis-(4-(oc temperature was gradually increased to 205' C. over tylamino)-1-pyridinium octane dibromide there was 2-3 hours and then maintained for 7 hours. The reaction obtained 4.30 g. of the corresponding dichloride, m.p. mixture was allowed to cool and approximately 2 liters 210-21 1 C, of boiling water was added as soon as the dark melt EXAMPLE 13 began to crystallize. The resulting solution was cooled, 10 Following a procedure similar to that described in made alkaline with 35% sodium hydroxide and ex Example 9D but employing 11.1 g (0.054 mole) of tracted with dichloromethane. The organic extracts 4-(octylamino)pyridine and 6.6 g (0.027 mole) of 1,6- were washed with water, dried over anhydrous sodium dibromohexane and heating the reaction mixture 9 sulfate and evaporated to dryness. The residual solid hours under reflux there was obtained 15.1 g of 1,6-bis was treated with 700 ml of boiling hexane, cooled, 15 4-(octylamino)-1-pyridinium hexane dibromide, m.p. filtered and washed with cold hexane to give 641 g of 136-138 C, air-dried 4-(octylamino)pyridine, mp. 66'-69 C. D. To a stirred, warm solution containing 100 g. EXAMPLE 14 (0.049 mole) of 4-(octylamino)pyridine in 150 ml of Following a procedure similar to that described in acetonitrile was added dropwise a solution containing 20 Example 2 but employing 5.0 g of 1,6-bis-(4-(oc 7.4 g (0.0245 mole) of 1,10-dibromodecane in 50 ml of tylamino)-1-pyridinium hexane dibromide there was acetonitrile and the resulting mixture heated 18 hours obtained 4.23g of the corresponding dichloride, m.p. under reflux. After cooling and stirring 1 hour at room 189-191 C. temperature, the precipitated solid was collected by EXAMPLE 1.5 filtration, washed with acetonitrile and dried 48 hours at 25 85 C. to give 15.6 g. of 1,10-bis-(4-(octylamino)-1- To a stirred warm solution containing 2.06 g. (0.01 pyridinium)decane dibromide, m.p. 163-164 C. mole) of 4-(octylamino)pyridine in 15 ml. of acetonitrile was added dropwise a solution containing 1.37 g (0.005 EXAMPLE 10 mole) of 1,6-hexanediol dimethanesulfonate in 10 ml. of 30 acetonitrile and the resulting mixture was heated under A. Following a procedure similar to that described in reflux 20 hours. The reaction mixture was evaporated to Example 2 but employing 5.0 g of 1,10-bis-(4-(oc dryness under reduced pressure and the residual gum tylamino)-1-pyridinium decane dibromide there was was triturated with ether to give a colorless hygro obtained 4.31 g of the corresponding dichloride, m.p. scopic solid. This product was redissolved in a mixture 213.214 C. 35 of ethanol, benzene and acetonitrile and the resulting B. Alternatively, a mixture containing 61.8 g. of 4 solution evaporated to dryness under reduced pressure. (octylamino)pyridine and 31.5 g. of 1,10-dichlorodec The residue was triturated with ether and the resulting ane was stirred and heated slowly to 120° C. The heat white solid was quickly collected by filtration, washed source was removed and the temperature of the now with anhydrous ether, and dried 72 hours at 28 C./0.1 exothermic reaction continued to rise to 180 C. As mm. to give 2.65 g. of 1,6-bis(4-(octylamino)-1-pyridini soon as the reaction mixture began to crystallize 250 ml. umhexane dimethanesulfonate as a waxy, white solid. of N,N-dimethylformamide was rapidly added and the resulting mixture was heated to give a clear homogene EXAMPLE 16 ous solution and then cooled to 0° C. The precipitated Following a procedure similar to that described in product was collected by filtration, washed with ether 45 Example 9D but employing 14.24 g (0.08 mole) of and dried 24 hours under vacuum at 60° C. to give 73 g. 4-(hexylamino)pyridine and 8.64 g (0.04 mole) of 1,4- of 1,10-bis-(4-(octylamino)-1-pyridinium decane dichlo dibromobutane there was obtained following trituration ride, m.p. 215-217 C. of the crude product with a mixture of acetonitrile and C. To a stirred, filtered solution containing 9.8 g. acetone 20.45 g. of 1,4-bis-(4-(hexylamino)-1-pyridini (0.0157 mole) of the above dichloride in 200 ml. of 50 umbutane dibromide, mp. 199-201 C. distilled water under nitrogen was added dropwise 14.9 EXAMPLE 17 g. (equivalent to 5.96 g. or 0.04 mole) of 40% sodium Following a procedure similar to that described in 1-oxo-2-pyridinethiolate. The resulting precipitate was Example 9D but employing 10.7 g (0.06 mole) of 4 collected, washed with water, and dried over phospho 55 (hexylamino)pyridine and 7.32 g (0.03 mole) of 1,6- rus pentoxide under vacuum affording 10.7 g of 1,10 dibromohexane there was obtained following tritura bis-(4-(octylamino)-1-pyridinium)decane bis(1-oxo-2- tion of the crude product with a mixture of ether, aceto pyridinethiolate) as a pale yellow solid, m.p. 134-137 nitrile and acetone 14.90 g. of 1,6-bis-(4-(hexylamino)-1- C. pyridinium-hexane dibromide, m.p. 178-179 C. 60 EXAMPLE 11 EXAMPLE 18 Following a procedure similar to that described in Following a procedure similar to that described in Example 9D but employing 11.1 g (0.054 mole) of Example 9D but employing 10.7 g (0.06 mole) of 4 4-(octylamino)pyridine and 7.34 g. (0.027 mole) of 1,8- (hexylamino)pyridine and 7.75 g (0.03 mole) of 1,7- dibromooctane, and heating the reaction mixture 6 65 dibromoheptane there was obtained following tritura hours under reflux there was obtained 15.6 g. of 1,8-bis tion of the crude product with a mixture of acetonitrile 4-(octylamino)-1-pyridinium octane dibromide, m.p. and acetone 16.4 g of 1,7-bis-4-(hexylamino)-1- 174-175° C. pyridiniumheptane dibromide, m.p. 157-158 C. 4,206,215 17 18 there was obtained 4.35g. of the corresponding dichlo EXAMPLE 9 ride, mp. 209-210' C. Following a procedure similar to that described in Example 9D but using 10.7 g (0.06 mole) of 4-(hex EXAMPLE 28 ylamino)-pyridine and 8.6 g (0.03 mole) of 1,9- 5 To a stirred slurry of 30 ml. of synthetic anion ex dibromononane there was obtained 17.15 g of 1.9-bis change resin in the hydroxide form (sold by Rohm and 4-(hexylamino)-1-pyridinium-nonane dibromide, m.p. Haas under the tradename Amberlite IRA 400) in 150 114-115 C. ml. of water was added dropwise 48% aqueous hydro fluoric acid until the mixture was acidic. After stirring EXAMPLE 20 10 an additional 0.5 hour the slurry was poured into a Following a procedure similar to that described in column. The column was drained and the resin washed Example 9D but employing 15.4 g (0.08 mole) of 4 with a solution containing 15 ml. of 48% aqueous hy (heptylamino)pyridine and 8.64 g (0.04 mole) of 1,4- drofluoric acid in 185 ml. of distilled water. The resin dibromobutane there was obtained 23.1 g of 1,4-bis-(4- was then washed with distilled water until the eluate (heptylamino)-1-pyridiniumbutane dibromide, m.p. 15 was weakly acidic, and then successively with 20%, 229'-230 C, 40%, and 50% aqueous methanol and finally with abso lute methanol until the eluate was neutral. To the top of EXAMPLE 2 this column of ion exchange resin now in the fluoride Following a procedure similar to that described in form was added a solution containing 0.25 g (0.000365 Example 9D but employing 10.0 g (0.052 mole) of 20 mole) of 1,10-bis-(4-heptylamino)-1-pyridinium)decane 4-(heptylamino)pyridine and 6.7 g (0.026 mole) of 1,7- dibromide in 1 ml. of methanol. Five fractions of 15 ml. dibromoheptane there was obtained 14.05 g. of 1,7-bis each were collected. The first three fractions were com 4-(heptylamino)-1-pyridinium]heptane dibromide, m.p. bined and evaporated to dryness under reduced pres 142-143' C, sure. The oily residue was dissolved in a mixture of 25 toluene and ethanol and the resulting solution evapo EXAMPLE 22 rated to dryness under reduced pressure. The residual Following a procedure similar to that described in oil was redissolved in a mixture of benzene and acetone Example 9D but employing 11.6 g. (0.06 mole) of 4 and the solution concentrated to a small volume. The (heptylamino)pyridine and 8.2 g (0.03 mole) of 1,8- solid which separated was collected by filtration and dibromoctane there was obtained after recrystallization 30 dried 24 hours at 24' C/0.1 mm. to give 0.11 g of from acetonitrileether 18.6 g. of 1,8-bis-(4-(hep impure 1,10-bis-(4-(heptylamino)-1-pyridinium)decane tylamino)-1-pyridinium-octane dibromide, m.p. difluoride, mp. 85-90 C. 161-162 C. EXAMPLE 29 EXAMPLE 23 35 A. A solid mixture containing 115 g (0.5 mole) of Following a procedure similar to that described in N-(4-pyridyl)pyridinium chloride hydrochloride and Example 2 but employing 5.0 g (0.0076 mole) of 1,8- 119 g (0.66 mole) of n-nonylamine hydrochloride was bis(4-(heptylamino)-1-pyridinium)octane dibromide heated in an oil bath up to a bath temperature of 220 C. there was obtained 4.1 g of the corresponding dichlo (internal temperature of 190-194° C) and the resulting ride, mp. 206-208 C. 40 liquid was stirred at that temperature for 2 hours. The reaction mixture was then cooled to 80 C., diluted with EXAMPLE 24 1,2 liters of ice and water, made alkaline with 35% Following a procedure similar to that described in aqueous sodium hydroxide and extracted with chloro Example 9D but employing 15.4 g (0.08 mole) of 4 form. The chloroform extracts were dried over anhy (heptylamino)pyridine and 11.44 g of 1,9-dibromono 45 drous sodium sulfate, treated with decolorizing carbon, nane there was obtained 21.3 g of 1.9-bis-(4-(hep filtered and evaporated to dryness under reduced pres tylamino)-1-pyridiniumnonane dibromide, m.p. sure. The residual viscous oil was cooled to -78 C. 115-116 C, and triturated with ether. The resulting solid was col lected by filtration and washed with cold ether. The EXAMPLE 25 filtrate afforded a second crop of solid. The combined Following a procedure similar to that described in crops were redissolved in chloroform and the resulting Example, 2 but employing 5.0 g (0.0075 mole) of 1,9- solution treated with decolorizing carbon and filtered. bis(4-(heptylamino)-1-pyridinium)nonane dibromide This was repeated once more and the filtrate was then there was obtained 4.2 g of the corresponding dichlo evaporated to dryness under reduced pressure. The ride, mp. 154'-155.C. 55 residual semi-solid was triturated with ether and cooled to give a nearly colorless solid which was collected by EXAMPLE 26 filtration, washed with cold ether and dried. The solid Following a procedure similar to that described in was taken up in chloroform and the resulting solution Example 9D but employing 15.4 g (0.08 mole) of 4 treated with decolorizing carbon, filtered and evapo (heptyl-amino)pyridine and 12.0 g (0.04 mole) of 1,10 rated to dryness under reduced pressure. Cooling the dibromodecane there was obtained 25.7g of 1,10-bis semi-solid so-obtained and trituration with ether af 4-(heptylamino)-1-pyridinium)decane dibromide, m.p. forded a colorless solid which was collected by filtra 163-165 C...... tion, washed with cold ether and dried to give 51.7 g of 4-(nonylamino)pyridine, mp. 59-60 C. The filtrate EXAMPLE 27 65 afforded a second crop of 11.6 g., m.p. 55-57 C. . Following a procedure similar to that described in B. Alternatively 4-(nonylamino)pyridine was pre Example 2 but employing 5.0 g (0.0073 mole) of 1,10 pared as follows: A mixture containing 39.5 g (0.42 bis-(4-(heptylamino)-1-pyridinium)decane dibromide mole) of 4-aminopyridine, 175 g. (1.24 moles) of nonyl 4,206,215 19 2O aldehyde, 5.0 g of 10% palladium-on-carbon hydroge 4-(hexylamino)-1-pyridinium)decane dibromide, m.p. nation catalyst and sufficient absolute ethanol to give a 148-149 C. total volume of 600 ml. was warmed and hydrogenated under an initial hydrogen pressure of 45 psi until the EXAMPLE 34 absorption of hydrogen ceased. The reaction mixture 5 Following a procedure similar to that described in was filtered to remove the catalyst and the filtrate was Example 29C but employing 13.4 g (0.076 mole) of evaporated to dryness under reduced pressure. The 4-(cyclohexylamino)pyridine and 8.2 g (0.038 mole) of residual oil was then vacuum distilled to remove any 1,4-dibromobutane there was obtained 18.2g. of 1,4-bis nonyl alcohol which may have been produced. A frac 4-(cyclohexylamino)-1-pyridiniumbutane dibromide, tion boiling at 63-64' C/4.5 mm. was collected and set O m.p. 288-290 C. aside. The residue was triturated with ether and cooled at -78 C. The solid which precipitated was collected EXAMPLE 35 by filtration and washed with cold ether. This product Following a procedure similar to that described in was dissolved in chloroform, the resulting solution Example 29C but employing 13.4 g (0.076 mole) of treated with decolorizing carbon, filtered and the fil 15 4-(cyclohexylamino)pyridine and 8.75 g (0.038 mole) of trate evaporated to dryness. The residue was triturated 1,5-dibromopentane there was obtained 19.0 g of 1,5- with ether and cooled to -78 C. The solid thus bis-(4-(cyclohexylamino)-1-pyridinium pentane dibro obtained was collected by filtration, washed with cold mide, m.p. 255-256 C. ether and air-dried to give 27.0 g of 4-(nonylamino)- EXAMPLE 36 pyridine, m.p. 57-59 C. 20 C. A stirred suspension of 11.0 g (0.05 mole) of 4 Following a procedure similar to that described in (nonylamino)pyridine in 150 ml. of acetonitrile was Example 29C but employing 13.4 g (0.076 mole) of warmed until a clear homogenous solution was ob 4-(cyclohexylamino)pyridine and 9.3 g (0.038 mole) of tained. To this clear solution was added dropwise a 1,6-dibromohexane there was obtained 19.1 g of 1,6-bis solution containing 6.8 g. (0.025 mole) of 1,8- 25 4-(cyclohexylamino)-1-pyridinium hexane dibromide, dibromooctane in 50 ml. of acetonitrile and the resulting m.p. 307-308 C. mixture heated 19 hours under reflux during which time EXAMPLE 37 a solid precipitated from solution. After cooling, the Following a procedure similar to that described in solid was collected by filtration, redissolved in metha 30 Example 29C but employing 13.4 g (0.076 mole) of nol, the resulting solution treated with decolorizing 4-(cyclohexylamino)pyridine and 9.8 g. of (0.038 mole) carbon, filtered and evaporated to dryness under re of 1,7-dibromoheptane there was obtained 2.01 g of duced pressure. Trituration of the residual oil with ether 1,7-bis-(4-(cyclohexylamino)-1-pyridiniumheptane di containing a small amount of acetonitrite produced a bromide, m.p. 311-313 C. colorless crystalline solid which was collected by filtra 35 tion and dried to give 15.5g of 1,8-bis-(4-(nonylamino)- EXAMPLE 38 1-pyridiniumloctane dibromide, m.p. 178-179' C. Following a procedure, similar to that described in Example 29C but employing 13.4 g (0.076 mole) of EXAMPLE 30 4-(cyclohexylamino)pyridine and 10.34 g (0.038 mole) Following a procedure similar to that described in 40 of 1,8-dibromooctane there was obtained 19.1 g of Example 29C but employing 3.54 g (0.026 mole) of 1,8-bis-(4-(cyclohexylamino)-1-pyridinium.octane di 4-(propylamino)pyridine and 3.90 g. (0.013 mole) of bromide, m.p. 270-271 C. 1,10-dibromodecane there was obtained 5.19 g, of 1,10 bis-(4-(propylamino)-1-pyridinium decane dibromide, EXAMPLE 39 m.p. 206-207 C. 45 Following a procedure similar to that described in Example 29C but employing 13.4 g (0.076 mole) of EXAMPLE 31 4-(cyclohexylamino)pyridine and 10.8 g. (0.038 mole) of Following a procedure similar to that described in 1,9-dibromononane there was obtained 16.3 g of 1,9- Example 29C but employing 14.24 g (0.08 mole) of bis-(4-(cyclohexylamino)-1-pyridiniumnonane dibro 4-(hexylamino)pyridine and 9.20 g (0.04 mole) of 1,5-50 mide, m.p. 149-151 C. dibromopentane there was obtained following recrys tallization from acetonitrile-acetone 12.3 g of 1,5-bis-(4- EXAMPLE 40 (hexylamino)-1-pyridinium pentane dibromide, m.p. Following a procedure, similar to that described in 155-156 C. . . . Example 29C but employing 13.4 g (0.076 mole) of 55 4-(cyclohexylamino)pyridine and 11.4g. (0.038 mole) of EXAMPLE 32 1,10-dibromodecane there was obtained 19.0 g of 1,10 Following a procedure similar to that described in bis-(4-(cyclohexylamino)-1-pyridinium decane dibro Example 29C but employing 10.7 g (0.06 mole) of 4 mide, m.p. 226-227 C. l (hexylamino)pyridine and 8.2 g (0.03 mole) of 1,8- dibromooctane there was obtained 16.3 g of 1,8-bis-(4- EXAMPLE 41 (hexylamino)-1-pyridinium octane dibromide, m.p. A. A mixture containing 298.0 g (1.33 moles) of N-(4- 180-181 C. pyridyl)pyridinium chloride hydrochloride and 322 g. (2 moles) of 2-ethylhexylamine hydrochloride was EXAMPLE 33 heated 2 hours with stirring in an oil bath at a bath Following a procedure similar to that described in 65 temperature of 215 C. The mixture was cooled to 60' Example 29C but employing 12.5 g. (0.07 mole) of 4 C., diluted with 500 ml. of water and kept cold by the (hexylamino)pyridine and 10.5 g (0.035 mole) of 1,10 addition of ice while, being made alkaline with 35% dibromodecane there was obtained 16.0 g of 1, 10-bis aqueous sodium hydroxide. The alkaline mixture was 4,206,215 21 22 extracted with ether and the ethereal extracts were cold acetonitrile. Two recrystallizations from acetoni dried over anhydrous sodium sulfate and evaporated to trile afforded 970 g. of 1,12-bis-(4-(2-ethylhexylamino)- dryness. The residual oil was distilled under reduced 1-pyridiniumdodecane dichloride, m.p. 168-171 C. pressure to give 101.0 g of 4-(2-ethylhexylamino)pyri B. Alternatively, by following a procedure similar to dine b.p. 145-150° C./0.9 mm. B. Alternatively 4-(2-ethylhexylamino)pyridine was that described in Example 2 but employing 3.0 g. prepared as follows: (0.00405 mole) of 1,12-bis-(4-(2-ethylhexylamino)-1- To a stirred solution containing 800 g. (8.4 moles) of pyridinium-dodecane dibromide there was obtained 2.0 4-aminopyridine and 1500 ml. of triethylamine in 6.41. g. of the corresponding dichloride, m.p. 172-173° C. of dichloromethane there was added over 3 hours a 10 EXAMPLE 43 solution containing 1610 g. (10.0 moles) of 2-ethylhex anoyl chloride in 1.6l. of dichloromethane. Throughout Following a procedure similar to that described in the addition the temperature was maintained at 15 C. Example 41C but employing 10.3 g (0.05 mole) of 4 When the addition was complete the mixture was (octylamino)pyridine and 6.45 g (0.025 mole) of 1,7- warmed on a steam bath 2 hours. After cooling the 15 dibromoheptane there was obtained 14.85g. of 1,7-bis reaction mixture was washed thoroughly with water, 4-(octylamino)-1-pyridiniumheptane dibromide, m.p. dried over anhydrous sodium sulfate, treated with de 129-131 C. colorizing carbon and filtered. Evaporation of the fil trate afforded 1843 g of N-(4-pyridyl)-2-ethylhexana EXAMPLE 44 mide. 20 A. Following a procedure similar to that described in To a mixture containing 100 g. (2.63 moles) of lithium Example 41C but using 11.1 g (0.054 mole) of 4-(oc aluminum hydride in 2 l. of tetrahydrofuran was added tylamino)pyridine and 7.72 g (0.027 mole) of 19 at a sufficient rate to maintain gentle reflux a solution dibromononane there was obtained 17.0 g of 1,9-bis-(4- containing 570 g. (2.62 moles) of N-(4-pyridyl)-2-ethyl (octylamino)-1-pyridiniumnonane dibromide, m.p. hexanamide in 4 l. of tetrahydrofuran. When the addi 25 117-119 C. tion was complete (approximately 3 hours) the reaction mixture was heated under reflux 7 hours. After cooling, B. The corresponding dichloride prepared according the mixture was treated successively with 100 ml. of to the procedure of Example 2 had a m.p. 161-162 C. water, 100 ml. of 15% aqueous sodium hydroxide and EXAMPLE 45 300 ml. of water. The solids were removed by filtration 30 and the solvent was evaporated from the filtrate under Following a procedure similar to that described in reduced pressure. The residual oil was combined with Example 41C but employing 10.3 g (0.05 mole) of 4 the product of a duplicate run and vacuum distilled to (octylamino)-pyridine and 8.2 g (0.025 mole) of 1,12 give 837 g. of 4-(2-ethylhexylamino)pyridine b.p. dibromododecane there was obtained 15.2 g of 1,12-bis 135-160 C/0.2 mm. 35 4-(octylamino)-1-pyridiniumdodecane dibromide, C. To a stirred, warm solution containing 10.3 g (0.05 mp. 119-120° C. mole) of 4-(2-ethylhexylamino)pyridine in 50 ml. of acetonitrile was added dropwise a solution containing EXAMPLE 46 8.2 g (0.025 mole) of 1,12-dibromodecane in 170 ml. of Following a procedure similar to that described in acetonitrile and the resulting mixture was heated 20 40 Example 41C but using 5.8 g. (0.028 mole) of 4-(oc hours under reflux. Upon cooling to 0° C. a first crop of tylamino)pyridine and 5.0 g (0.014 mole) of 1,14 product precipitated and was cooled by filtration. dibromotetradecane there was obtained 9.3 g of 1,14 Evaporation of the filtrate and trituration of the residue bis-(4-(octylamino)-1-pyridiniumtetradecane dibro with ether afforded a second crop. The combined crops mide, m.p. 113'-115 C. were dissolved in methanol, the resulting solution 45 treated with decolorizing carbon, filtered and the fil EXAMPLE 47 trate evaporated to dryness under reduced pressure. Following a procedure simolar to that described in The residual oil was cooled and triturated with ether to Example 41C but employing 11.0 g (0.05 mole) of 4 give a slightly discolored solid. Recrystallization from (nonylamino)-pyridine and 6.1 g (0.025 mole) of 1,6- acetonitrile-ether followed by trituration of the product 50 with ether followed by acetonitrile afforded, after dry dibromohexane there was obtained 15.2 g of 1,6-bis-(4- ing 72 hours under vacuum at 60 C., 14.7 g of 1,12-bis (nonylamino)-1-pyridinium-hexane dibromide, m.p. 4-(2-ethylhexylamino)-1-pyridinium)dodecane dibro 152-154 C. mide as colorless granules, m.p. 146'14 147 C. EXAMPLE 48 EXAMPLE 42 55 Following a procedure similar to that described in A. A mixture containing 353.5 g. (1.72 mole) of 4-(2- Example 4 but employing 6.5 g. (0.0095 mole) of 1,6-bis ethylhexylamino)pyridine and 205 g (0.86 mole) of 4-(nonylamino)-1-pyridinium hexane dibromide there 1,12-dichlorododecane was heated to 120' C. The heat was obtained 4.95 g. of the corresponding dichloride, source was removed and the temperature of the now 60 m.p. 194-195 C. exothermic reaction continued to rise to 180-190° C. When the temperature dropped to 135° C. one liter of EXAMPLE 49 acetonitrile was carefully added and the mixture heated Following a procedure similar to that described in under reflux to give a clear solution. The hot acetoni Example 41C but employing 8.8 g. (0.04 mole) of 4 trile solution was combined with similar solutions from 65 (nonylamino)pyridine and 5.2 g (0.02 mole) of 1,7- two other runs, treated with decolorizing carbon and dibromoheptane there was obtained 12.2 g of 1,7-bis-(4- filtered. The filtrate was cooled and the precipitated (nonylamino)-1-pyridinium heptane dibromide, m.p. product was collected by filtration and washed with 132-134' C. 4,206,215 23 24 EXAMPLE 50 EXAMPLE 57 Following a procedure similar to that described in Following a procedure similar to that described in Example 41C but employing 11.0 g (0.05 mole) of 4 Example 53 but employing 12.4 g (0.06 mole) of 4-(2- (nonylamino)pyridine and 7.15 g (0.025 mole) of 1,9- 5 ethylhexylamino)pyridine and 8.6 g. (0.03 mole) of 1,9- dibromononane there was obtained 15.7g of 1.9-bis-(4- dibromononane there was obtained 15.2 g of 1.9-bis-(4- (nonylamino)-1-pyridinium nonane dibromide, m.p. (2-ethylhexylamino)-1-pyridiniumnonane dibromide, 121-122 C. mp. 158-159 C. EXAMPLE 5 10 EXAMPLE S8 Following a procedure similar to that described in Following a procedure similar to that described in Example 41C but employing 11.0 g (0.05 mole) of 4 Example 53 but employing 12.4 g (0.06 mole) of 4-(2- (nonylamino)pyridine and 7.5 g. (0.025 mole) of 1,10 ethylhexylamino)pyridine and 9.0 g (0.03 mole) of 1,10 dibromodecane there was obtained 15.63 g of 1,10-bis dibromodecane there was obtained 17.4 g of 1,10-bis 4-(nonylamino)-1-pyridinium)decane dibromide, m.p. 15 4-(2-ethylhexylamino)-1-pyridinium)decane dibro 172-173° C. mide, m.p. 162-163 C, EXAMPLE 52 EXAMPLE 59 Following a procedure similar to that described in Following a procedure similar to that described in Example 41C but employing 10.12 g (0.046 mole) of 20 Example 2 but employing 5.0 g of 1,10-bis-(4-(2-ethyl 4-(nonylamino)pyridine and 7.54 g (0.023 mole) of 1,12 hexylamino)-1-pyridinium decane dibromide there was dibromododecane there was obtained 16.4g. of 1,12-bis obtained 4.1.1 g of the corresponding dichloride, m.p. 4-(nonylamino)1-pyridiniumdodecane dibromide, m.p. 191-192 C. 105-106 C. EXAMPLE 60 EXAMPLE 53 25 To a stirred warm solution containing 12.0 g (0.063 To a stirred, warm solution containing 12.4 g (0.06 mole) of 4-(heptylamino)pyridine in 100 ml. of acetoni mole) of 4-(2-ethylhexylamino)pyridine in 100 ml. of trile there was added dropwise a solution containing 7.4 acetonitrile was added dropwise a solution containing g. (0.032 mole) of 1,5-dibromopentane in 25 ml. oface 6.9 g (0.03 mole) of 1,5-dibromopentane in 25 ml. of 30 tonitrile and the resulting mixture heated 19 hours acetonitrile and the resulting solution heated 20 hours under reflux. The reaction mixture was cooled in ice under reflux. The reaction mixture was cooled and and ether was gradually added until a colorless solid diluted with ether until slightly cloudy. Further cooling precipitated. The solid was collected by filtration re and stirring afforded a solid precipitate which was col crystallized from acetonitrile-ether and dried 48 hours lected by filtration and washed with a cold mixture of 35 at 90° C./1 mm. to give 15.9 g of 1,5-bis-(4-(hep acetonitrile and ether. The solid thus-obtained was dis tylamino)-1-pyridiniumpentane dibromide, m.p. solved in ethanol and the resulting solution treated with 153-154 C. decolorizing carbon and filtered. Evaporation of the filtrate provided a pale yellow viscous oil which was EXAMPLE 61 crystallized from acetonitrile-ether. The resulting solid Following a procedure similar to that described in was collected by filtration, washed with cold acetoni Example 60 but using 19.2 g (0.1 mole) of 4-(heptyl)- trile-ether and dried 24 hours under vacuum at 90 C. to amino)pyridine and 12.2 g (0.05 mole) of 1,6- give 13.6 g. of 1,5-bis-4-(2-ethylhexylamino)-1- dibromohexane there was obtained 27.5 g. of crude pyridiniumlpentane dibromide, m.p. 150-151 C. product. Recrystallization of a 15-gram sample from 45 acetonitrile-ether afforded 11.45 g. of 1,6-bis-4-(hep EXAMPLE 54 tylamino)-1-pyridinium hexane dibromide, m.p. Following a procedure similar to that described in 149-150 C. Example 53 but employing 12.4 g (0.06 mole) of 4-(2- ethylhexylamino)pyridine and 7.32 g (0.03 mole) of EXAMPLE 62 1,6-dibromohexane there was obtained 16.1 g of 1,6-bis- 50 A. A suspension of 24.0 g of crude 1,6-bis-4-(hep 4-(2-ethylhexylamino)-1-pyridinium hexane dibro tylamino)-1-pyridinium hexane dibromide in 500 ml. of mide, m.p. 208-209 C. water was made alkaline with 3 N aqueous sodium hydroxide and extracted with chloroform. The chloro EXAMPLE 55 form extracts were dried over anhydrous sodium sulfate Following a procedure similar to that described in 55 and evaporated to dryness under reduced pressure. The Example 53 but employing 12.4 g (0.06 mole) of 4-(2- residual oil was dissolved in methanol and the resulting ethylhexylamino)pyridine and 7.75 g (0.03 mole) of solution acidified with methanesulfonic acid and evapo 1,7-dibromoheptane there was obtained 17.9 g of 1,7- rated to dryness under reduced pressure. The residue bis-4-(2-ethylhexylamino)-1-pyridiniumheptane dibro was redissolved in methanol, treated with decolorizing mide, m.p. 219–220 C. 60 carbon, filtered and evaporated to dryness under vac uum leaving an oily residue which upon trituration with EXAMPLE 56 ether followed by acetonitrile provide 18.2 g of gummy Following a procedure similar to that described in solid. The crude solid was dissolved in water and the Example 53 but employing 12.4 g (0.06 mole) of 4-(2- resulting solution make alkaline with 35% aqueous so ethylhexylamino)pyridine and 8.2 g (0.03 mole) of 1,8- 65 dium hydroxide and extracted with chloroform. The dibromooctane there was obtained 15.9 g, of 1,8-bis-(4- chloroform extracts were dried over anhydrous sodium (2-ethylhexylamino)-1-pyridinium octane dibromide, sulfate and evaporated to dryness under reduced pres m.p. 160-161° C. sure. The resulting solid was triturated successively 4,206,215 25 26 with ether and acetonitrile, collected by filtration and was cooled by the addition of ice and made alkaline dried to give 16.3.g. of tan solid, m.p. 105-108' C. The with 200 ml. of 35% aqueous sodium hydroxide. The solid so-obtained was dissolved in 100 ml. of methanol dark solid which separated was collected by filtration and the resulting solution acidified with methanesul and washed with cold water. This material was dis fonic acid. Evaporation to dryness under reduced pres solved in 1 1. of chloroform and the resulting solution sure produced a viscous oil which was taken up in 20 was treated with decolorizing carbon and filtered. The ml. of methanol and treated portionwise with 150 ml. of filtrate was evaporated under reduced pressure and the water. The resulting suspension was cooled in ice and residue was triturated with 150 ml. of ether. The solid the suspended solid was collected by filtration and thus-obtained was redissolved in chloroform and the washed with cold water. The solid was then slurried in 10 resulting solution treated with decolorizing carbon and hot acetone, cooled, collected by filtration, washed filtered. The filtrate was again treated with decolorizing with cold acetone and dried 48 hours at 95 C/1 mm. to carbon and filtered. The filtrate was evaporated to dry give 10.8 g. of tan solid, m.p. 163'-165 C. This product ness under reduced pressure and the residual solid was gave a positive chloride ion test with silver nitrate and triturated with ether. Recrystallization from acetoni the nmr spectrum.thereof indicated the absence of the trile afforded after drying under vacuum 96.1 g of methanesulfonate group. 4-(decylamino)pyridine, m.p.71-73' C. B. A suspension of 14.0 g of 1,6-bis-(4-(heptylamino)- B. To a stirred, warm solution containing 12.2 g. 1-pyridinium hexane dibromide in 500 ml. of water was (0.052 mole) of 4-(decylamino)pyridine in 150-170 ml. made alkaline with 35% aqueous sodium hydroxide of acetonitrile was added dropwise a solution contain and extracted with chloroform. The chloroform ex 20 ing 6.35 g (0.026 mole) of 1,6-dibromohexane in 60 ml. tracts were dried over anhydrous sodium sulfate and of acetonitrile and the resulting mixture heated approxi evaporated to dryness under reduced pressure. The mately 20 hours under reflux. The reaction mixture was partially solid residue was dissolved in 1 l. of acetoni then cooled in ice and the precipitated solid was col trile-benzene and the resulting solution evaporated to lected by filtration and washed with cold acetonitrile. dryness under vacuum. This process was repeated three 25 The product thus-obtained was dissolved in methanol times. The final residue was dissolved in 50 ml. of aceto and the resulting solution treated with decolorizing nitrile and the resulting solution diluted with 50 ml. of carbon and filtered. The filtrate was evaporated to dry benzene and cooled in ice to give after filtering and ness under reduced pressure and the residue was tritu drying 5.3 g of tan solid. The filtrate afforded a second rated with 50 ml, of cold ether, collected by filtration crop of 4.8 g. The crops were combined and dissolved 30 and dried 2 days over phosphorus pentoxide at 70 in 50 ml. ofacetonitrile. Dilution with ether precipitated C/0.1 mm. to give 14.6 g. of 1,6-bis-(4-(decylamino)-1- a solid which was collected by filtration and dried 48 pyridiniumhexane dibromide, mp. 138-140 C. hours at 95 C/1 mm. to give 8.85g. of tan solid, m.p. 174'-176' C. This product also gave a positive chloride EXAMPLE 6S ion test with silver nitrate. 35 Following a procedure similar to that described in C. The products of parts A and B above were com Example 64B but employing 12.2 g (0.052 mole) of bined, mixed with 300 ml. of water and heated until a 4-(decylamino)pyridine and 6.7 g (0.026 mole) of 1,7- homogeneous solution was obtained. The solution was dibromoheptane there were obtained 16.0 g of 1,7-bis filtered and the filtrate treated with 50 ml. of 12 N hy 4-(decylamino)-1-pyridiniumheptane dibromide, m.p. drochloric acid. The resulting suspension was concen 145-147 C. trated under reduced pressure and the residue was treated with ice. The solid so-produced was collected EXAMPLE 66 by filtration, washed with cold water and dried. The Following a procedure similar to that described in product was then dissolved in acetone, the resulting Example 64B but employing 11.7 g (0.050 mole) of solution diluted with a mixture of benzene and ethanol, 45 4-(decylamino)pyridine and 6.8 g. (0.025 mole) of 1,8- and the whole evaporated to dryness under vacuum. dibromooctane there was obtained 16.9 g of 1,8-bis-(4- The solid residue was slurried in 100 ml of acetone and (decylamino)-1-pyridinium.octane dibromide, m.p. the slurry diluted with ether and filtered to give after 180°-182 C. drying 48 hours at 105 C/1 mm. and 72 hours at 115 C/1 mm. 16.3 g of crude 1,6-bis-(4-(heptylamino)-1- SO EXAMPLE 67 pyridinium hexane dichloride as an off-white solid, m.p. Following a procedure similar to that described in 176-178 C. Example 64B but employing 11.7 g (0.050 mole) of 4-(decylamino)pyridine and 7.15 g. (0.025 mole) of 1,9- EXAMPLE 63 dibromononane there was obtained 15.1 g of 1.9-bis-(4- Following a procedure similar to that described in 55 (decylamino)-1-pyridiniumnonane dibromide, m.p. Example 60 but employing 10.30 g. (0.050 mole) of 124-126 C. 4-(2-ethylhexylamino)pyridine and 5.4 g (0.025 mole) of 1,4-dibromobutane there was obtained 14.9 g, of EXAMPLE 68 1,4-bis-(4-(2-ethylhexylamino)-1-pyridiniumbutane di Following a procedure similar to that described in bromide, m.p. 245'-246' C. 60 Example 64B but employing 11.2 g (0.048 mole) of 4-(decylamino)pyridine and 7.2 g (0.024 mole) of 1,10 EXAMPLE 64 dibromodecane there was obtained 14.6 g. of 1,10-bis A. A mixture containing 229 g (1.0 mole) of N-(4- 4-(decylamino)-1-pyridinium)decane dibromide, m.p. pyridyl)pyridinium chloride hydrochloride and 244 g. 173-174 C, (1.26 moles) of n-decylamine hydrochloride was stirred 65 and heated approximately 2.5 hours at 190-195' C. The EXAMPLE 69 reaction mixture was then allowed to cool slowly to 40 Following a procedure similar to that described in C. and diluted with 2.l. of water. The resulting solution Example 64B but employing 11.2 g (0.048 mole) of 4,206,215 27 28 4-(decylamino)pyridine and 7.9 g (0.024 mole) of 1,12 mixture was heated 2.5 hours on a steam bath. The dibromododecane there was obtained 16.5 g. of 1,12-bis reaction mixture was cooled to room temperature, di 4-(decylamino)-1-pyridiniumdodecane dibromide, luted with ether and the product allowed to crystallize. m.p. 102-106 C. The precipitated solid was collected by filtration, washed with a mixture of acetonitrile and ether and EXAMPLE 70 air-dried to give 27.4 g of 1,4-bis-(4-(p-chloro To a stirred warm solution containing 13.6 g. (0.052 phenylamino)-1-pyridinium)butane dibromide, mp. mole) of 4-(decylamino)pyridine in 140 ml. of acetoni 182-189 C. trile was added dropwise a solution containing 6.34 g. B. A suspension of the above product in 500 ml. of (0.026 mole) of 1,6-dibromohexane in 50 ml. of acetoni O water was treated with ice and 2 N aqueous sodium trile and the resulting mixture heated under reflux over hydroxide and the resulting mixture extracted thor night. The reaction mixture was cooled and the precipi oughly with chloroform. The chloroform extracts were tated solid was collected by filtration. The collected dried over anhydrous sodium sulfate and evaporated to solid was dissolved in absolute ethanol and the resulting dryness under reduced pressure to give 21.0 g of the solution treated with decolorizing carbon and filtered. 15 corresponding anhydro base as an oil which solidified The filtrate was evaporated to dryness under reduced on standing. pressure affording a residual white solid which was C. A solution containing the above anhydro base in slurried in ether, collected by filtration and dried at 60 250 ml. of methanol was acidified with a solution of C./0.1 mm. to give 17.63 g. of 1,6-bis-(4- methane-sulfonic acid in methanol. Evaporation to dry (dodecylamino)-1-pyridiniumhexane dibromide, m.p. 20 ness under reduced pressure afforded an oil which was 164-165 C. crystallized from methanol-ether. The solid thus obtained was recrystallized from methanol-ether to give EXAMPLE 71 after drying 48 hrs. at 92 C./0.1 mm. 18.95g. of 1,4-bis Following a procedure similar to that described in 4-(p-chlorophenylamino)-1-pyridiniumbutane dime Example 70 but employing 13.6 g. (0.052 mole) of 4- 25 thanesulfonate, m.p. 245-247 C. (dodecylamino)pyridine and 6.71 g (0.026 mole) of 1,7-dibromoheptane there was obtained 18.03 g. of 1,7- EXAMPLE 77 bis-(4-(dodecylamino)-1-pyridinium)heptane dibromide, A. To a stirred, warm solution containing 21.0 g. m.p. 148-150 C. (0.102 mole) of 4-(p-chlorophenylamino)pyridine in 200 30 ml. of N,N-dimethylformamide was added dropwise a EXAMPLE 72 solution containing 12.45 g (0.051 mole) of 1,6- Following a procedure similar to that described in dibromohexane in 50 ml. of acetonitrile and the result Example 70 but employing 12.59 g (0.048 mole) of ing mixture was heated 4 hours on a steam bath. Upon 4-(dodecylamino)pyridine and 6.53 g (0.024 mole) of cooling to room temperature a solid began to precipi 1,8-dibromooctane there was obtained 16.18 g. of 1,8-35 tate. The reaction mixture was diluted with 150 ml of bis-(4-(dodecylamino)-1-pyridinium octane dibromide, acetonitrile and cooled further in ice. The precipitated m.p. 184-185° C. solid was collected by filtration and air-dried to give 27.2 g of 1,6-bis-(4-(p-chlorophenylamino)-1-pyridini EXAMPLE 73 umhexane dibromide, m.p. 148-150° C. Following a procedure similar to that described in 40 B. A methanol solution of the corresponding anhydro Example 70 but employing 12.59 g (0.048 mole) of base prepared from the above dibromide according to 4-(dodecylamino)pyridine and 6.86 g. (0.024 mole) of the procedure of Example 76B was acidified with meth 1,9-dibromononane there was obtained 19.35 g. of 1,9- anesulfonic acid and then evaporated to dryness under bis-(4-(dodecylamino)-1-pyridiniumnonane dibromide, reduced pressure. The residual oil was crystallized from m.p. 134-135 C. 45 acetone-methanol to give after drying 48 hrs. at 75 C/0.1 mm. 25.2 g of 1,6-bis-(4-(p-chlorophenylamino)- EXAMPLE 74 1-pyridinium hexane dimethanesulfonate, m.p. Following a procedure similar to that described in 108°-110° C. - Example 70 but employing 12.59 g (0.048 mole) of 4-(dodecylamino)pyridine and 7.2 g (0.024 mole) of 50 EXAMPLE 78 1,10-dibromodecane there was obtained 18.08 g. of 1,10 A. To a stirred, warm solution containing 21.0 g. bis-(4-(dodecylamino)-1-pyridinium decane dibromide, (0.102 mole) of 4-(p-chlorophenylamino)pyridine in 200 mp. 178°-180 C. ml. of N,N-dimethylformamide was added dropwise a solution containing 13.9 g (0.051 mole) of 1,8- EXAMPLE 75 55 dibromooctane in 50 ml. of acetonitrile and the resulting Following a procedure similar to that described in mixture heated 1.5 hours on a steam bath. The reaction Example 70 but employing 11.54 g. (0.044 mole) of mixture was then diluted with 100 ml. of acetonitrile 4-(dodecylamino)pyridine and 7.22 g (0.022 mole) of and heating was continued another 2.5 hours when an 1,12-dibromododecane there was obtained 17.68 g. of additional 100 ml. of acetonitrile was added. After heat 1,12-bis-(4-(dodecylamino)-1-pyridiniumdodecane di- 60 ing another 2 hours the reaction mixture was cooled and bromide, mp. 75-77 C. the solid which had precipitated was collected by filtra tion, washed with a mixture of acetonitrile and ether EXAMPLE 76 and dried to give 30.1 g of 1,8-bis-(4-(p-chloro A. To a stirred warm solution containing 21.0 g. phenylamino)-1-pyridinium octane dibromide, m.p. (0.102 mole) of 4-(p-chlorophenylamino)pyridine in 150 65 245-247 C. - ml. of N,N-dimethylformamide was added dropwise a B. A methanol solution of the corresponding anhydro solution containing 11.02 g (0.051 mole) of 1,4- base prepared from the above dibromide according to dibromobutane in 50 ml. of acetonitrile and the resulting the procedure of Example 76B was acidified with meth 4,206,215 29 30 anesulfonic acid and then evaporated to dryness under under reduced pressure and the residual oil crystallized reduced pressure. The residual oil was dissolved in from ethanol-acetonitrile. The solid so-produced was acetonitrile and the resulting solution treated with ace dissolved in methanol and the resulting solution treated tone until cloudy followed by clarification with a small with decolorizing carbon and filtered. The filtrate was amount of methanol and cooled. The crude solid which evaporated to dryness under vacuum and the residual precipitated was collected and recrystallized from ace oil again crystallized from ethanol-acetonitrile. The tonitrile acetone to give after drying 36 hours at 80 product was collected by filtration and dried 36 hours at C./0.1 mm., 23.5 g. of 1,8-bis-(4-(p-chloro 105' C/0.1 mm. to give 10.4 g of 1,7-bis-(4-(p-chloro phenylamino)-1-pyridinium octane dimethanesulfonate, phenylamino)-1-pyridinium]heptane dibromide, m.p. m.p. 164-165 C. 10 202-204' C. EXAMPLE 79 EXAMPLE 82 A. To a stirred, warm solution containing 21.0 g. Following a procedure similar to that described in (0.102 mole) of 4-(p-chlorophenylamino)pyridine in 200 Example 81 but employing 10.0 g (0.049 mole) of 4-(p- ml. of N,N-dimethylformamide was added dropwise a 15 chlorophenylamino)pyridine and 7.15g. (0.025 mole) of solution containing 15.3 g (0.051 mole) of 1,10 1,9-dibromononane there was obtained 11.2 g of 1,9- dibromodecane in 50 ml. of acetonitrile and the result bis-(4-(p-chlorophenylamino)-1-pyridiniumnonane di ing mixture was heated 3.5 hours on a steam bath. The bromide, mp. 178-179 C. reaction mixture was then evaporated to dryness under reduced pressure and the residual oil was crystallized 20 EXAMPLE 83 from methanol-acetonitrile. The solid so-produced was A. A mixture containing 64.0 g (0.43 mole) of p collected by filtration and washed with a cold mixture fluoroaniline hydrochloride and 4-chloropyridine hy of acetonitrile and ether to give 28.0 g of 1,10-bis-4-(p- drochloride was heated gradually to an internal temper chlorophenylamino)-1-pyridinium)decane dibromide, ature of 170° C. whereupon the solid mixture began to m.p. 225-230 C. 25 melt and stirring was begun. Upon continued heating, B. A methanol solution of the corresponding anhydro the melt began to resolidify. Stirring was stopped and base prepared from the above dibromide according to heating was continued until no further change was evi the procedure of Example 76B was acidified with meth dent. The mixture was cooled, dissolved in 1 liter of anesulfonic acid and then evaporated to dryness under water and ice, and the solution made alkaline with 35% reduced pressure. The residual oil was crystallized from 30 sodium hydroxide. The resulting precipitate was col acetonitrile-ether. The crude solid thus-obtained was lected, washed with water and redissolved in hot meth dissolved in methanol and the resulting solution was anol. The methanol solution was treated with decoloriz treated with decolorizing carbon and filtered. Evapora ing carbon, filtered and evaporated to dryness. The tion of the filtrate afforded an oil which was suspended residue was recrystallized twice from chloroform in a mixture of acetonitrile and acetone to give a crude 35 acetonitrile-ether to give after drying 47.3 g of 4-(p- solid. Recrystallization from methanol-ether afforded fluorophenylamino)pyridine, m.p. 195'-198 C. after drying 48 hours at 95 C/0.1 mm. 18.4 g of 1,10 B. To a warm, stirred solution containing 7.5 g (0.04 bis-(4-(p-chlorophenylamino)-1-pyridinium)decane mole) of 4-(p-fluorophenylamino)pyridine in a mixture dimethanesulfonate, m.p. 202-204 C. of 75 ml. of acetonitrile and 25 ml. of N,N-dimethylfor mamide was added dropwise over 0.25 hour 4.9 g, EXAMPLE 80 (0.020 mole) of 1,6-dibromohexane. The resulting mix To a stirred warm solution containing 10.0 g (0.049 ture was heated 18 hours under gentle reflux, and then mole) of 4-(p-chlorophenylamino)pyridine in a mixture cooled to 0°C. The solution was seeded with crystals of of 275 ml. of acetonitrile and 100 ml. of N,N-dimethyl the desired product (previously obtained by diluting a formamide was added dropwise a solution containing 45 small aliquot of the reaction mixture with ether) and the 5.75 g (0.025 mole) of 1,5-dibromopentane in 25 ml. of resulting precipitate was collected and washed with acetonitrile and the resulting mixture heated 24 hours cold ether. The solid was dissolved in methanol and the under reflux. The reaction mixture was then evaporated resulting solution was treated with decolorizing carbon, to dryness under reduced pressure and the residue tritu filtered and evaporated to dryness. The residue was rated with a mixture of ether and acetonitrile. The pale 50 recrystallized from methanol-acetonitrile-ether to give yellow solid so-obtained was redissolved in ethanol and 9.3 g of 1,6-bis-(4-(p-fluorophenylamino)-1-pyridini the resulting solution treated with decolorizing carbon umhexane dibromide, m.p. 220-222 C. and filtered. The filtrate was evaporated to dryness under reduced pressure and the residual oil was crystal EXAMPLE 84 lized from ether-acetonitrile. The colorless solid was 55 Following a procedure similar to that described in recrystallized from methanolacetonitrile and dried 48 Example 83B but employing 7.5 g (0.04 mole) of 4-(p- hours at 115 C/0.1 mm. to give 8.1 g of 1,5-bis-(4-(p- fluorophenylamino)pyridine and 5.4 g (0.020 mole) of chlorophenylamino)-1-pyridinium)pentane dibromide, 1,8-dibromooctane there was obtained 8.65g. of 1,8-bis m.p. 166-168 C. 4-p-fluorophenylamino)-1-pyridinium octane dibro 60 mide, m.p. 226-228 C. EXAMPLE 81 By following procedures similar to those described in To a stirred, warm solution containing 10.0 g (0.049 the foregoing examples, the following bis-(4-(R-amino)- mole) of 4-(p-chlorophenylamino)pyridine in a mixture 1-pyridiniumalkanes are obtained: of 125 ml. of N,N-dimethylformamide and 50 ml. of 3-ethyl-1,6-bis-(4-(tetradecylamino)-1-pyridinium acetonitrile was added dropwise a solution containing 65 hexane dibromide by reacting 1,6-dibromo-3-ethylhex 6.45 g. of 1,7-dibromoheptane in 25 ml. of acetonitrile ane with 4-(tetradecylamino)pyridine which is in turn and the resulting mixture heated 19 hours under reflux. obtained by reacting N-(4-pyridyl)pyridinium chloride The reaction mixture was then evaporated to dryness hydrochloride with tetradecylamine hydrochloride, 4,206,215 31. 32 1,5-bis-(4-(octadecylamino)-1-pyridiniumlpentane obtained by reacting N-(4-pyridyl)pyridinium chloride dibromide by reacting 1,5-dibromopentane with 4-(oc hydrochloride with p-anisidine hydrochloride, tadecylamino)pyridine which in turn is obtained by 1, 11-bis-(4-(m-nitrophenylamino)-1-pyridinium reacting N-(4-pyridyl)pyridinium chloride hydrochlo undecane dibromide by reacting 1,11-dibromoundecane ride with octadecylamine hydrochloride, with 4-(m-nitrophenylamino)pyridine which in turn is 1, 11-bis-(4-(2,2-dimethylbutylamino)-1-pyridinium obtained by reacting N-(4-pyridyl)pyridinium chloride 3-methylundecane dibromide by reacting 1,11-dibromo hydrochloride with m-nitroaniline hydrochloride, 3-methyl-undecane with 4-(2,2-dimethylbutylamino)- 1,12-bis-(4-(o-cyanophenylamino)-1-pyridinium pyridine which in turn is obtained by reacting N-(4- dodecane dibromide by reacting 1,12-dibromododecane pyridyl)pyridinium chloride hydrochloride with 2,2- 10 with 4-(o-cyanophenylamino)pyridine which in turn is dimethylbutylamine hydrochloride, obtained by reacting N-(4-pyridyl)pyridinium chloride 1, 18-bis-(4-(1,4-dimethylpentylamino)-1-pyridinium)- hydrochloride with o-cyanoaniline hydrochloride, octadecane dibromide by reacting 1,18-dibromoocta 1,16-bis-<4-m-(trifluoromethyl)phenylamino)-1- decane with 4-(1,4-dimethylpentylamino)pyridine pyridinium) hexadecane dibromide by reacting 1,16 which in turn is obtained by reacting N-(4-pyridyl)- 15 dibromohexadecane and 4-m-(trifluoromethyl)- pyridinium chloride hydrochloride with 1,4-dimethyl phenylaminolpyridine which in turn is obtained by re pentylamine hydrochloride, acting N-(4-pyridyl)pyridinium chloride hydrochloride 4,9-dimethyl-1,12-bis-(4-(1,5-dimethyl-4-ethyl-hex with m-(trifluoromethyl)aniline hydrochloride, ylamino)-1-pyridiniumdodecane dibromide by reacting 1,7-bis-(4-(2-methoxy-5-methylphenylamino)-1- 1,12-dibromo-4,9-dimethyldodecane with 4-(1,5- 20 pyridinium]heptane dibromide by reacting 1,7- dimethyl-4-ethylhexylamino)pyridine which in turn is dibromoheptane with 4-(2-methoxy-5-methyl obtained by reacting N-(4-pyridyl)pyridinium chloride phenylamino)pyridine which in turn is obtained by hydrochloride with 1,5-dimethyl-4-ethylhexylamine reacting N-(4-pyridyl)pyridinium chloride hydrochlo hydrochloride, ride with 2-methoxy-5-methylaniline hydrochloride, 1,10-bis-(4-(cyclopentylamino)-1-pyridinium-decane 25 1,10-bis-2-methyl-4-(octylamino)-1-pyridinium-dec dichloride by reacting 1,10-dichlorodecane with 4-(cy ane dibromide by reacting 1,10-dibromodecane with clopentylamino)pyridine which in turn is obtained by 2-methyl-4-(octylamino)pyridine which is in turn ob reacting N-(4-pyridyl)pyridinium chloride hydrochlo tained by reacting 4-chloro-2-methylpyridine hydro ride with cyclopentylamine hydrochloride, chloride with n-octylamine hydrochloride, 1,12-bis-(4-(cycloheptylamino)-1-pyridinium-dodec 30 1,8-bis-(3-ethyl-4-(heptylamino)-1-pyridinium ane dibromide by reacting 1,12-dibromododecane with octane dibromide by reacting 1,8-dibromooctane with 4-(cyclopheptylamino)pyridine which in turn is ob 3-ethyl-4-(heptylamino)pyridine which is in turn ob tained by reacting N-(4-pyridyl)pyridinium chloride tained by reacting 4-chloro-3-ethylpyridine hydrochlo hydrochloride with cycloheptylamine hydrochloride, ride with n-heptylamine hydrochloride, 1,6-bis-(4-(p-bromophenylamino)-1-pyridinium-3- 35 1,10-bis-(4-(octylamino)-1-pyridinium-1-methyldec methylhexane dibromide by reacting 1,6-dibromo-3- ane dibromide by reacting 1,10-dibromo-1-methyldec methylhexane with 4-(p-bromophenylamino)pyridine ane with 4-(octylamino)pyridine. which in turn is obtained by reacting N-(4-pyridyl)- The following are further illustrative examples of pyridinium chloride hydrochloride with p-bromoani bis-(4-(R-amino)-1-pyridinium)alkanes of Formula I line hydrochloride, which are obtained in accordance with the above 1,10-bis-4-(m-fluorophenylamino)-1-pyridinium described procedures: decane dibromide by reacting 1,10-dibromodecane with 3-methyl-1,5-bis-(4-(undecylamino)-1-pyridinium 4-(m-fluorophenylamino)pyridine which in turn is ob pentane dibromide, 3-propyl-1,5-bis-(4-(tridecylamino)- tained by reacting N-(4-pyridyl)pyridinium chloride 1-pyridinium pentane dichloride, 4,4-dimethyl-1,7-bis hydrochloride with m-fluoroaniline hydrochloride, 45 4-(pentadecylamino)-1-pyridinium]heptane sulfate, 2,6- 1,4-bis-(4-(3-chloro-4-fluorophenylamino)-1-pyridini dimethyl-1,7-bis-(4-(hexadecylamino)-1-pyridinium um)-2-ethylbutane dibromide by reacting 1,4-dibromo heptane disulfamate, 1-methyl-1,4-bis-(4-(hep 2-ethylbutane with 4-(3-chloro-4-fluorophenylamino)- tadecylamino)-1-pyridiniumbutane dibenzenesulfonate, pyridine which in turn is obtained by reacting N-(4- 2,4,4-trimethyl-1,6-bis-(4-(2-methylhexylamino)-1- pyridyl)pyridinium chloride hydrochloride with 3 50 pyridinium hexane bis-cyclohexylsulfamate, 1,17-bis-(4- chloro-4-fluoroaniline hydrochloride, (3-ethylpentylamino)-1-pyridinium-heptadecane dibro 1,10-bis-4-(benzylamino)-1-pyridinium decane di mide, 1,16-bis-4-(1-ethylhexylamino)-1-pyridinium chloride by reacting 1,10-dichlorodecane with 4-(ben hexadecane dinitrate, 1,15-bis-4-(heptylamino)-1- zylamino)pyridine, pyridinium)pentadecane dibromide, 1,13-bis-(4-(oc 1,8-bis-(4-(3,4-methylenedioxyphenylamino)-1- 55 tylamino)-1-pyridiniumtridecane di-2-naphthalenesul pyridinium]octane dibromide by reacting 1,8- fonate, 1,11-bis-4-(3-ethylheptylamino)-1-pyridinium dibromooctane with 4-(3,4-methylenedioxy undecane 2,6-naphthalenedisulfonate, -4-methyl-1,14 phenylamino)pyridine which in turn is obtained by bis-4-(2-propylpentylamino)-1-pyridiniumtetradecane reacting N-(4-pyridyl)pyridinium chloride hydrochlo dichloride, 5-ethyl-1,9-bis(4-(1,3,5-trimethylhex ride with 3,4-methylenedioxyaniline hydrochloride, ylamino)-1-pyridinium nonane dibromide, 3,3,6,6,-tet 1.9-bis-4-(p-ethylphenylamino)-1-pyridinium nonane ramethyl-1,8-bis-(4-(heptylamino)-1-pyridinium-octane dibromide by reacting 1,9-dibromononane with 4-(p- dibromide, 2,13-dimethyl-1,14-bis-4-(1,2-dimethyl-tet ethylphenylamino)pyridine which in turn is obtained by radecylamino)-1-pyridinium tetradecyl dibromide, 1,6- reacting N-(4-pyridyl)pyridinium chloride hydrochlo bis(4-(1-methylpentylamino)-1-pyridinium hexane di-p- ride with p-ethylaniline hydrochloride, 65 toluenesulfonate, 1,8-bis-(4-(2-methylheptylamino)-1- 1,10-bis-(4-(p-methoxyphenylamino)-1-pyridinium pyridinium octane diiodide, 1,8-bis-(4-(2-methyl-3- decane dichloride by reacting 1,10-dichlorodecane with ethylpentylamino)-1-pyridinium]octane diethanesulfon 4-(p-methoxyphenylamino)pyridine which is in turn ate, 1,9-bis-(4-(o-chlorophenylamino)-1-pyridinium no 4,206,215 33 34 nane dibromide, 1,8-bis-4-(p-iodophenylamino)-1- cup of the Autotray is added 0.1 ml. of the test solution. pyridinium)octane diiodide, 1,12-bis-(4-(2,4-difluoro Activation of the Autotiter initiates a sequence of oper phenylamino)-1-pyridiniumdodecane dibromide, 1,11 ations by which 0.05 ml. of the test compound solution bis-(4-(2,5-dibromophenylamino)-1-pyridinium undec is withdrawn from the cup by a Microtiter transfer loop ane, dibromide, 1,10-bis-(4-(3,5-dichlorophenylamino)- and diluted in 0.05 ml, of sterile water. Following this 1-pyridinium-decane dichloride, 1,8-bis-(4-(2- operation, 0.05 ml, of inoculated double-strength semi fluorobenzylamino)-1-pyridinium]octane dichloride, synthetic medium (glucose) is added automatically to 1,18-bis-(4-(3,4-dichlorobenzylamino)-1-pyridinium)oc each cup. The overall operation results in final drug tadecane dibromide, 1,4-bis-(4-(4-hydroxy-3-methox concentrations ranging from 500 to 0.06 mcg/ml. in ybenzylamino)-1-pyridinium)butane dibromide, 1,12 10 twofold decrements. The Autotray is incubated for bis-(4-(4-methylbenzylamino)-1-pyridiniumdodecane 18-20 hours at 37 C. at which time the trays are exam dibromide, 1,12-bis-3-isopropyl-4-(hexylamino)-1- ined visually for growth as evidenced by turbidity, and pyridinium)dodecane dichloride, 1.9-bis-(2-n-butyl-4- the concentration of the last sample in the series show (nonylamino)-1-pyridinium nonane dibromide, 1,12-bis ing no growth (or no turbidity) is recorded as the mini 4-(2-ethylhexylamino)-2-methyl-1-pyridiniumdodec 5 mal inhibitory concentration (MIC) in mcg/ml. The ane dibromide, 1,8-bis-(4-(cyclohexylamino)-2-methyl compounds were thus tested as solutions against a vari 1-pyridinium octane dibromide and 1.9-bis-(4-(p- ety of gram positive and gram negative bacteria includ chlorophenylamino)-2-methyl-1-pyridiniumnonane di ing Staphylococcus aureus, Proteus mirabilis, Escherichia bromide. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, As indicated hereinabove and exemplified in Table A 20 Streptococcus pyogenes, and against such fungi as Asper hereinbelow, the novel compounds of Formula I have gillus niger, Candida albicans and Trichophyton menta useful antibacterial and antifungal activity being effec grophytes, tive against both gram positive and gram negative bac Many of the compounds of Formula I were also teria, and several species of fungi. Of particular impor found to be antibacterially effective against Streptococ tance is the fact that the compounds are highly active 25 cus mutans and Actinomyces A-viscosis, against Pseudomonas aeruginosa and Proteus mirabilis, The dental plaque preventive activity of certain of two ubiquitous gram negative bacteria which are often the compounds of the present invention was determined the pathogens associated with hospital-acquired infec by measuring the ability of these compounds to inhibit tions and which are often resistant to other currently the production of dental plaque by Streptococcus mutans used antibacterial agents. Thus the present compounds 30 OMZ-6 as follows: are true broad spectrum antibacterial agents. Moreover, A culture medium for the plaque-producing Strepto these compounds are virucidally effective against Her coccus mutans OMZ-61 containing 1.5 g. of BBL beef pes viruses. extract, 5 g. of sodium chloride, 10 g. of dehydrated By comparison, the most closely related prior art trypticase, 5.g. of sucrose and sufficient distilled water compounds (compounds I-IX in Table A), i.e. 35 to give a total volume of 1000 ml. is adjusted to pH 7.0 I. 1,8-bis-(4-amino-1-pyridinium)octane dibromide, and sterilized by membrane filtration. The medium is II. 1,10-bis-(4-amino-1-pyridinium)decane dibromide, dispensed aseptically in 10-ml. aliquots into 150x16 III. 1,4-bis-(4-amino-1-pyridinium)butane dibromide, mm. test tubes and stored at refrigeration temperature IV. 1,6-bis-(4-amino-1-pyridinium)hexane dibromide, until used. V. 1,12-bis-(4-amino-1-pyridinium)dodecane dibro Two concentrations of the compound to be tested are mide, prepared by dissolving 100 mg. of the compound in 1 VI. 1,10-bis-(4-amino-1-pyridinium)-1-methyldecane ml. of distilled water with the aid of sufficient 0.1 N dibromide, sodium hydroxide, 10% dimethylsulfoxide or 10% VII. 1,9-bis-(4-amino-1-pyridinium)nonane dibro N,N-dimethylformamide and diluting the resulting solu mide, 45 tion to 10 ml, with distilled water. This 1.0% solution VIII. 1,8-bis-(4-amino-2-methyl-1-pyridinium)octane and a 1:10 dilution in distilled water (0.1%) are steril dibromide, and ized by membrane filtration before use. IX. 1.9-bis-(4-amino-2-methyl-1-pyridinium)nonane Asterile piece of plaque-free natural tooth enamel or dibromide, synthetic hydroxylapatite is suspended in each concen are relatively weakly active, have a narrow spectrum of 50 tration of compound for two 1-minute periods, each activity and are essentially inactive against both Pseudo followed by a 1-minute air drying period. Each piece is monas aeruginosa and Proteus mirabilis. Nevertheless, then suspended and agitated for 5 minutes in individual the discovery of this relatively weak and specific anti test tubes containing sterile distilled water (rinse). They microbial activity is considered a part of the present are then suspended in 10 ml. of liquid beef extract me invention and accordingly the method of using the 55 dium to which has been added 0.3 ml of a 24-hr. anaero known compounds as antimicrobial agents is claimed bic culture of Streptococcus mutans. The tubes contain herein. However, it is clear that the novel claimed com ing the "treated” hydroxylapatite and the Streptococcus pounds of Formula I possess marked advantages over mutans are then incubated anaerobically at 37 C. for 24 the known compounds both in terms of potency and hours. The same process of two 1-minute soaks in the spectrum of activity and are accordingly believed unob solution of compound, each followed by 1 minute of air vious over and patentably distinct from the known drying and the final 5-minute rinse is repeated before compounds. once again suspending the hydroxylapatite in a fresh The antibacterial and antifungal activity of represen tube containing 10 ml. of beef extract medium and 0.3 tative examples of the compounds of Formula I was ml. of inoculum. At the end of the second 24-hour per determined using a modification of the Autotiter 65 iod, each piece of hydroxylapatite is rinsed for 1 minute method described by Goss, et al. Applied Microbiology in 3 successive tubes of distilled water. These are then 16 (9) 1,414-1,416 (1968) in which a 1,000 mcg/ml. suspended for 1 minute in a solution of F, D & C Red solution of the test compound is prepared. To the first No. 3 dye. This staining procedure is used in order to 4,206,215 35 36 identify more easily the development of plaque after the ing concentrations for representative examples are re 48-hour period of exposure to the plaque-producing ported in Table Chereinbelow. organism. The staining period is followed by another Antibacterial effectiveness in the presence of serum 10-second rinse to remove excess dye. Any plaque for was determined for representative examples by the mation stains a brilliant pink. Test results are read as standard serial tube dilution test wherein paired com plaque inhibition (active) or no plaque inhibition (inac parisons of minimum inhibitory concentrations (MIC) tive) at the percent concentration tested. Active com were determined in the absence and in the presence of pounds are tested at successively lower doses in order heat inactivated (30 min. at 56' C.) horse serum (40%) to determine the minimum effective concentration. On in the test medium. The results are expressed as the fold some occasions, the interference of plaque production is 10 increase in MIC (mcg/ml.) versus Staphyloccocus au caused by the inhibition of growth of the organism in reus and Escherichia coli caused by the presence of the culture medium because the compound has been serum and are presented in Table B hereinbelow. leached from the hydroxylapatite to produce an anti Dental plaque-preventive effectiveness in the pres bacterial level in the medium. When this occurs, the ence of sterile human saliva (50%) was determined for compound is tested at lower concentrations until the 15 representative examples. Test results are read as mini growth of the organism in the surrounding medium is mum effective concentration and are presented in Table equal to that in the nonmedicated control culture. Chereinbelow. Plaque may or may not be formed at these lower con Virucidal activity of representative examples against centrations. Herpes simplex type 2 virus was determined in vitro Substantivity of the test compound to the tooth sur 20 employing a procedure similar to the plaque inhibition face which is of course necessary to prevent the forma test for detection of specific inhibitors of DNA-contain tion of dental plaque thereon, may, however, potentiate ing viruses published by E. C. Herrmann, Jr., Proc. Soc. the accumulation of staining agents, e.g., coffee, to Exp. Biol. Med. 107, 142 (1961) wherein 2 mg. of the bacco, food colorants, etc., on the tooth surface. There test compound was placed on the surface of the growth fore the above procedure was also utilized to estimate 25 medium. The compounds corresponding to Examples staining potential, i.e., the ability of the test compound 1D, 3, 5C, 7, 9D, 11, 13, 17, 18, 21, 22, 24, 31, 33, 35, 36, to promote adherence of staining agents to the tooth 37, 53, 54, 55,56, 57,58, 60, 61, 63, 81 and 82 were found surface. Thus, a residual, uniform pale pink color adher active and the compounds of Examples 16, 34, 39, 40 ing to the test surface following the final rinse is indica and 79B were found inactive. tive of staining potential at the precent concentration 30 The acute oral toxicity (ALD50) of several represen tested and is reported as staining concentration. The tative examples was determined as follows: Groups of 3 uniform pale pink color caused by the test compound is young adult male Swiss-Webster strain albino mice readily distinguished from the localized, brilliant pink were fasted approximately 4 hours before medication color caused by plaque formation. In general the pre and then medicated once orally by stomach tube. All ferred species advantageously have a staining concen 35 mice were observed for 7 days after medication. Au tration considerably higher than the minimum effective topsy of all mice in the study revealed no gross tissue plaque-preventive concentration. changes except for the group receiving 1000 mg/kg. of Numerical antibacterial, antifungal, and dental the compound of Example 25 wherein congestion of the plaque-preventive test data for the compounds are pres glandular portion of the stomachs of two of the three ented in Table A hereinbelow. Corresponding data for mice was observed. The results are presented in Table the above-identified known compounds I-IX are also D hereinbelow. included in Table A for purposes of comparison. Stain TABLE A In Vitro Antibacterial and Antifungal Activity MIC mcg/ml Plague Preventive Activity Concentration Cpd. of S. E. K. P. Ps, E. coli S. A. C. albi. C. albi- T. Tested (%) Ex. Aureus coli pneum. mirab. aerug. E. coli 100/ pyogen. niger s cans menta, Ac- Inac No. Smith Vogel 396.45 MGH-1 MGH-2 AB1932-1 B22 C203 6404 10231 Wisc. 9129 tive tive 76C 7.8 15.6 >250 >250 > 125 7.8 15.6 1.95 250 125 12.5 62.5 1.0 77B 3.9 15.6 500 500 S00 3.9 15.6 95 62.5 62.S. 2S 15.6 1.0 78B 0.4 3.1 3.3 125 125 1.6 3. 0.2 31.3 62.5 62.5 313 10 79B 0.4 1.6 3.1 > 125 62.5 0.8 .6 0.4 25 3.13 6.25 6.25 1.0 6 0.2 3.1 6.3 >250 125 1.6 1.6 0.4 25 6.25 6.25 : 3.13. O 62C 0.24, 3.9 3.9 > 25 >62.5 1.95 1.95 0.98 7.8 3.9 3.9 3.9 10 21 0.24, 1.95 3.9 125 125 .95 1.95 0.98, 5.6 3.9 3.9 3.9 10 60 0.49 31.3 3.3 125 125 3.9 7.8 1.95 15.6 15.6 5.6 7.8 1.0 22 0.39 .95 195 62.5 313 0.98 0.98 0.49 15.6 0.98 1.95 1.95 0.1 17 0.49 31.3 125 >250 > 25 7.8 7.8 78 250 7.8 15.6 O 8 0.39 5.6 15.6 > 25 > 125 7.8 7.8 0.49 15.6 6.25 7.8 5.6 O 23 0.5 1.95 .95 62.6 3.3 0 .95 1.0 7.8 1.0 . 19S 1.95 0. 32 0.24 3.9 7.8 >250 125 1.95 3.9 0.49 7.8 3.9 ; 3.9 7.8 O 19 0.39 1.95 3.9 25 125 1.95 .95 0.49 15.6 1.95 3.9 3.9. 1.0 33 0.195 1.95 3.9 31.3 31.3 0.98 1.95 0.49 3.9 95 3.9 1.95 10 31 .95 125 > 125 >250 > 125 3.3 62.5. 0.98. 31.3 313 62.5 31.3 10 16 7.8 125 > 125 > 125 >250 62.5 25 1.95 125 125 25 15.6 10 20 0.98 31.3 > 125 >250 >250 7.8 15.6 0.49 15.6 S.6 15.6 3.9 1.0 26 0.37 0.75 1.49 6.0 .9 0.75 0.75 1.95 2.98 0.75 1.49 0.75 0.01 24 0.25 1.0 1.95 5.6 62.5 O .95 0.5 3.9 1.0 1.0 0.5 0.003 4,206,215 37 38 TABLE A-continued In Vitro Antibacterial and Antifungal Activity MIC (ncg/ml Plague Preventive Activity Concentration Cpd. of S. E. K. P. Ps. E. coli S. A. C. albi. C. albi- T Tested (%) Ex. Aureus coli pneum, mirab. aerug. E. coli 100/ pyogen, niger Cas cans menta. Ac- Inac No. Smith Vogel 396.45 MGH-1 MGH-2 AB1932-1 B22 C203 6404 1023 Wisc. 929 tive tive 25 0.25 .0 1.95 31.3 31.3 10 1.0 0.5 1.95 1.0 .0 1.0 0004 63 0.98 62.5 > 125 > 125 125 7.8 31.3 1.25 31.3 31.3 31.3 15.6 1.0 53 0.49 15.6 > 125 > 125 125 7.8 15.6 0.625 31.3 31.3 15.6 7.8 10 54 0.49 7.8 125 > 125 >250 1.95 3.9 0.625 15.6 7.8 7.8 .95 1.0 55 0.49 3.9 7.8 > 125 125 3.9 3.9 0.49 15.6 7.8 7.8 1.95 1.0 57 0.31 0.98 7.8 125 25 0.98 0.98 0.625 5.6 3.9 3.9 0.98 1.0 27 0.25 0.5 1.0 7.8 7.8 1.0 0.5 0.5 3.9 1.96 1.0 1.0 0004 58 0.24 1.5 3.0 12.1 48.4 0.37 0.76 1.95 6.0 3.0 1.5 0.76 0.003 59 0.25 .0 .95 7.8 31.3 1.0 0.5 1.0 15.6 1.95 1.95 1.0 0003 56 0.3 3.9 7.8 125 125 0.98 1.95 0.6 5.6 3.9 3.9 1.95 0.004 81 0.98 3.9 > 125 >125 > 125 125 3.9 0.49 31.3 31.3 31.3 7.8 1.0 82 0.625 0.98 7.8 > 125 125 0.98 1.95 0.24 15.6 3.9 3.9 3.9 1.0 34 125 125 >500 >500 >500 125 125 31.3 500 500 500 125 1.0 35 125 >500 >500 >500 >500 d500 >500 15.6 500 500 500 125 1.0 36 15.6 & 125 > 125 > 125 > 125 125 > 125 15.6 25 125 125 5.6 1.0 37 7.8 & 125 > 125 > 125 25 125 > 125 7.8 > 125 25 125 5.6 1.0 80 7.8 125 > 25 > 125 > 125 3.3 125 1.9S 62.5 62.5 62.5 15.6 1.0 38 3.9 62.5 > 125 > 125 250 31.3 62.5 0.93 15.6 31.3 31.3 3.9 1.0 40 0.24, 15.6 > 125 > 125 > 125 7.8 15.6 3.9 7.8 3.9 7.8 1.95 1.0 39 0.98 31.3 125 >500 500 31.3 31.3 3.9 7.8 15.6 15.6 3.9 1.0 3 0.39 156 3.13 3.13 6.25 6.25 3.13 0.39 3.13 1.56 3.13 1.56 0004 2 10 1.95 3.9 3.9 1.95 1.0 .0 1.0 7.8 3.9 7.8 1.0 001 4. 0.25 0.5 1.95 1.95 3.9 1.0 0.5 1.0 7.8 1.95 .95 0.5 0.003 8 0.25 1.0 1.0 7.8 5.6 1.0 0.5 1.0 7.8 1.0 1.0 10 00 1D 0.77 1.54 0.77 54 3.0 3.0 0.77 1.95 3.0 1.54 1.54 0.77 0.1 10A,B 0.5 1.95 1.95 3.9 3.9 1.0 1.0 1.0 7.8 1.95 1.95 1.95 0.00S 7 0.15 0.37 1.5 6.0 23.8 0.37 0.37 1.0 3.0 0.74 0.74 0.37 0.01 SC 0.38 0.76 0.76 1.5 6.0 0.76 0.76 .0 3.0 1.5 0.76 0.76 0.005 6 0.5 1.0 1.0 3.9 3.9 0.5 0.5 1.95 3.9 .0 1.95 0.5 0.004 13 0.5 1.95 3.9 31.3 31.3 1.0 1.95 0.5 3.9 195 1.95 0.5 0.004 14 0.25 1.0 1.95 15.6 31.3 0.5 0.5 0.5 15.6 1.9S 1.95 1.0 0004 0.25 0 195 7.8 15.6 1.0 1.95 1.0 3.9 1.0 1.0 0.5 0.004 12 0.25 1.0 1.0 3.9 7.8 1.0 0.5 0.5 7.8 .0 1.0 1.0 0005 9D 0.5 .95 3.9 3.9 3.9 1.0 1.95 0.5 1.95 1.0 1.0 1.0 0004 45 0.5 1.95 1.0 .95 1.95 1.95 1.95 1.0 7.8 1.0 3.9 10 0005 43 0.5 1.95 1.0 7.8 3.3 1.0 1.0 10 7.8 1.95 1.95 0 0005 46 10 7.8 7.8 3.9 3.9 3.9 3.9 1.95 15.6 7.8 7.8 1.95 0005 44A 1.0 1.0 1.95 15.6 3.9 1.0 1.0 0.5 3.9 1.0 1.95 1.0 0003 47 0.5 1.0 3.9 3.9 15.6 1.95 1.95 1.0 15.6 3.9 1.95 10 0004 48 1.0 .95 3.9 7.8 15.6 1.95 1.95 1.0 3.9 1.95 195 1.0 00 29C 1.0 3.9 1.95 3.9 7.8 1.95 1.95 1.95 7.8 3.9 3.9 1.95 0004 50 1.95 1.95 3.9 3.9 3.9 3.9 1.95 1.95 7.8 3.9 3.9 1.95 0004 51 1.95 1.95 3.9 3.9 3.9 3.9 3.9 1.95 7.8 3.9 3.9 1.95 0.004 52 1.95 3.9 3.9 3.9 3.9 7.8 3.9 1.95 15.6 7.8 7.8 1.95 0.01 72 7.8 31.3 >62.5 62.5 >62.5 62.5 > 62.5 15.6 62.5 31.3 31.3 7.8 0. 73 7.8 62.5 >62.5 62.5 >62.5 - d. 62.5 62.5 15.6 d62.5 d 62.5 31.3 7.8 0.1 70 7.8 3.3 >62.5 & 62.5 > 62.5 & 62.5 62.5 7.8 62.5 15.6 31.3 7.8 0,1 71 3.9 62.5 >62.5 62.5 62.5 62.5 62.5 7.8 62.5 62.5 62.5 7.8 0.1 74 15.6 62.5 62.5 62.5 >62.5 62.5 >62.5 15.6 62.5 62.5 62.5 15.6 0.1 75 31.3 62.5 62.5 d62.5 >62.5 d62.5 >62.5 15.6 62.5 & 62.5 62.5 15.6 1.0 65 1.0 3.9 7.8 3.9 7.8 3.9 3.9 1.0 7.8 15.6 15.6 1.95 0.01 66 1.0 .95 3.9 7.8 7.8' 7.8 3.9 95 . 15.6 7.8 15.6 1.95 0.01 67 1.0 , 1.9S 15.6 7.8 7.8 7.8 3.9 1.9S 31.3 3.3 15.6 3.9 0.01 68 1.95 7.8 31.3 7.8 15.6 15.6 15.6 3.9 15.6 15.6 15.6 3.9 0.01 69 1.95 7.8 31.3 15.6 15.6 31.3 15.6 3.9 15.6 15.6 5.6 3.9 0.1 49 0.5 1.0 1.95 3.9 15.6 1.95 1.95 10 7.8 1.95 1.95 1.0 0004 64B 10 1.95 3.9 1.95 7.8 .95 7.8 1.0 7.8 1.95 3.9 1.0 001 4C 0.5 10 1.95 3.9 3.9 1.0 1.0 0.5 3.9 1.0 1.0 1.0 0.1 30 1.95 & 62.5 & 62.5 250 >62.5 62.5 > 62.5 3.9 3.3 15.6 7.8 1.0 1.0 10C 1.0 10 1.95 1.95 3.9 --- - 0.5 1.95 1.0 - 1.0 0005 83B 15.6 31.3 > 125 > 125 >62.5 15.6 15.6 7.8 31.3 31.3 31.3 31.3 1.0 84 1.95 15.6 >62.5 & 125 >62.5 7.8 15.6 7.8 15.6 15.6 7.8 15.6 1.0 I 31.3 125 >500 >500 >500 125 250 31.3 125 25 125 125 1.0 I 7.8 62.5 250 >500 500 31.3 62.5 7.8 125 250 62.5 15.6 1.0 III 125 250 >500 >500 >500 - - 62.5 500 >500 - 500 1.0 IV 125 250 >500 >500 >500 - 62.5 500 >500 - >500 .0 V 1.9S 62.5 > 62.5 d 125 >62.5 - -- 1.0 3.3 31.3 - 3.9 1.0 VI 15.6 250 500 >500 >500 - - 15.6 250 31.3 - 5.6 1.0 VII 31.3 250 500 >500 >500 - - 7.8 250 31.3 - 15.6 10 VIII 31.3 250 500 >500 >500 - - 7.8 125 25 - 5.6 1.0 IX 3.9 31.3 250 >500 >500 - - 3.9 31.3 31.3 - 7.8 1.0 4,206,215 39 40 TABLE B Table D-continued - Effect of Serum on Bacteriostatic Activity Acute Oral Toxicity (ALDs) - MIC (meg/ml) vs. Indicated Bacteria Cpd, of Solution or Dose, mg/kg, of Mortality S. Aureus ATCC 6538 E. Col ATCC 8739 5 Ex, No. Suspension Total Material 24 Hrs 7 Days Cpd. of 0% 40% Fol 0% 40% Fold 7-Day ALD50 = 1000 mg/kg Ex, No. Serum Serum Increase Serum Serum Increase 25 Aqueous 250 79B 0.25 7.8 32 0.5 5.6 32 Solution 500 22 0.25 3.9 16 . 0.5 3.9 8 000 3/3 3/3 33 0.125 1.9S 16 0.25 3.9 16 7-Day ALD50 = 500 mg/kg 26 0,25 1.9S 16 0.5 7.8 16 10 3 Suspension 125 Ov3 O/3 24 0,25 1.9S 16 0.25 7.8 32 in 19 G.T. 250 O/3 Ov3 58 0.25 3.9 16 0,5 S.6 32 500 O/3 3 0,39 3,2 8 0,78 25.0 32 1000 1 0,95 3,12, 16 1.56 25.0 16 7-Day ALD50 = 1000 mg/kg 7 O,O 156 16 0.39 12.5 32 4. Suspension 25 O/3 0/3 SC 0,10 3.2 32 0.78 25.0 32 15 in 12, G.T. 1250 O/3 O/3 3 0.25 31.2 128 10 31.2 32 500 O/3 O/3 11 O.S 7.8 16 OS 3.2 64 1000 0/3 0/3 9D 0.5 7.8 16 O 62.5 64 7-Day ALD50 = > 1000 mg/kg 29C O.S 7.8 16 3.9 62.5 6 2 Suspension 125 O/3 O/3 41C 0.25 7.8 32 O 62.S 64 in 12 G.T. 250 O/3 20 500 0/3 0/3 1000 0/3 7-Day ALD50 = 1000 mg/kg TABLE C 3 Aqueous 125 O/3 O/3 Staining Potential and Effect of Saliva on Solution 250 Dental Plaque Preventive Activity is: s 3. P Dental Plaque. 25 7-Day ALD50 = 315 mg/kg reventive Activity 5C S Cpd. of in 50% saliva Stainin Suspension 125 O/3 0/3 p ng in 2, G.T. 250 Ov3 0/3 Ex. No, Minimum Effective Conc. (%) Concentration (%) 500 O/3 0/3 27 0.05 1000 24 Ca. 0.005 0.05 7-Day ALD50 = > 1000 mg/kg 25 0.05 30 6 Aqueous 12S 58 Ca. 0.005 0.05 Solution 250 O/3 59 0.05 500 S6 0.1 0.05 1000 3/3 3/3 3 0.1 0.05 7-Day ALD50 = 315 mg/kg 4. Ca. 0.005 0.005 1D Suspension 125 O/3 O/3 5C 0.051 0.005 35 in 2, G.T. 250 0/3 0/3 6 0.005 500 : 13 0.005 1000 14 0, 0.05 7-Day ALD50 = 750 mg/kg 11 0.05 0.05 1. Suspension 125 O/3 O/3 12 0.05 in 12 G.T. 250 Ov3 0/3 9D Ca, 0.005 0.05 40 500 0/3 10 A, B 0.005 0.05 1000 O/3 0/3 44A 0.005 0.05 7-Day ALD50 = 1000 mg/kg 47 0.004 2 Aqueous 25 Ov3 Ov3 29C 0.004 Solution 250 O/3 0/3 50 0.004 500 51 0.004 45 1000 49 0.005 7-Day ALD50 = 750 mg/kg A. 9D Suspension 25 Ov3 0/3 No bacterial growth at this concentration, plaque formed at 0.005% in 19, G.T. 250 0/3 0/3 500 0/3 0/3 Table D 000 0/3 0/3 50 7-Day ALD50 = > 1000 mg/kg - Acute Oral Toxicity (ALD50) 10A,B Aqueous 125 O/3 O/3 Cpd. of Solution or Dose, mg/kg, of Mortality Solution 250 0/3 O/3 Ex. No. Suspensiono Total Material 24 Hrs 7 Days 1000500 O/3 0/3 23 Aqueous 125 0/3 Ov3 7-Day ALD50 = c 1000 mg/kg Solution 250 0/3 O/3 58 Suspension in 125 Ov3 O/3 500 55 12 G.T. 250 O/3 O/3 1000 3/3 3/3 500 Ov3 0/3 7-Day ALD50 = 625 mg/kg 1000 O/3 O/3 26 Suspension 25 Ov3 O/3 7-Day ALD50 = > 1000 mg/kg in 1% G.T. 250 0/3 Ov3 59 Aqueous 125 0/3 O/3 500 0/3 0/3 Solution 250 0/3 O/3 000 0/3 0/3 60 500 7-Day ALD50 = >1000 mg/kg 1000 27 Aqueous 25 Ov3 7-Day ALD50 = 750 mg/kg Solution 250 O/3 O/3 S6 Suspension in 125 O/3 Ov3 500 1% G.T. 250 O/3 0/3 000 3/3 500 O/3 O/3 7-Day ALD50 = 250 mg/kg 65 1000 24 Suspension 25 O/3 O/3 7-Day ALD50 = > 1000 mg/kg in 12 G.T. 250 0/3 O/3 14 Suspension in 125 O/3 0/3 500 O/3 12, GT. 250 O/3 0/3 1000 O/3 O/3 500 O/3 O/3 4,206,215 41 42 Table D-continued x is 1, 2 or 3; and wherein Acute Oral Toxicity (ALDSO) (m)(2)=(n)(x). Cpd. of Solution or Dose, mg/kg, of Mortality 3. A compound according to claim 2 wherein R1 is Ex. No. Suspension Total Material 24 Hrs 7 Days 5 hydrogen, Y is (CH2) and w is from 4 to 18. 1000 0/3 7-Day ALD50 = x 1000 mg/kg 4. A compound according to claim 3 wherein w is 44A Suspension in 125 0/3 0/3 from 8 to 12. 1% G.T. 250 0/3 0/3 5. A compound according to claim 4 wherein R is an 500 0/3 0/3 alkyl group containing from 7 to 9 carbon atoms. 1000 0/3 0/3 10 7-Day ALD50 = 1000 mg/kg 6. A compound according to claim 5 wherein A is 29C Suspension in 125 0/3 0/3 chloride or bromide. 1%. G.T. 250 0/3 0/3 7. A compound having the formula 500 Ov3 0/3 1000 O/3 0/3 7-Day ALD50 = 1000 mg/kg 15 41C Suspension in 125 0/3 O/3 W 7 --2 1% G.T. 2SO 0/3 0/3 RNH / Ne-Y-N W NHR 500 0/3 0/3 1000 O/3 0/3 - A* 7-Day ALD50 = x 1000 mg/kg R R G.T. = Gum Tragacanth 20 I claim: wherein: 1. A compound having the formula Y is an alkylene group containing from 4 to 18 carbon atoms and separating the two 4-(R-NH)-1- 25 pyridinyl groups by from 4 to 18 carbon atoms; 7 --2 W R is a cycloalkyl group containing from 5 to 7 carbon RNH / N-Y-N V NHR atoms; R1 is hydrogen or lower alkyl; : , , A* A is an anion; R R n 30 m is 1 or 3; n is 1 or 2; wherein: x is 1, 2 or 3; and Y is an alkylene group containing from 4 to 18 carbon wherein atoms and separating the two 4-(R-NH)-1- (m)(2)=(n)(x). pyridinyl groups by from 4 to 18 carbon atoms; 35 8. A compound according to claim 7 wherein R is R is an alkyl group containing from 6 to 18 carbon hydrogen, Y is (CH2) and w is from 4 to 18. atoms, a cycloalkyl group containing from 5 to 7 9. A compound according to claim 8 wherein w is carbon atoms, benzyl, or phenyl substituted by from 8 to 12 and R is cyclohexyl. methylenedioxy or one or two substituents selected 10. A compound having the formula from the group consisting of halogen, lower-alkyl, lower-alkoxy, nitro, cyano and trifluoromethyl; 7 --2 R1 is hydrogen or lower alkyl; W A is an anion; RNH / N-Y-N W NHR m is 1 or 3; Ain' n is 1 or 2; 45 x is 1, 2 or 3; and R R wherein (m)(2)=(n)(x). wherein: 2. A compound having the formula Y is an alkylene group containing from 4 to 18 carbon 50 atoms and separating the two 4-(R-NH)-1-pyridi nyl groups by from 4 to 18 carbon atoms; R is phenyl substituted by methylenedioxy or one or two substituents selected from the group consisting of halogen, lower-alkyl, lower-alkoxy, nitro, cyano and trifluoromethyl; R1 is hydrogen or lower alkyl; A is an anion; wherein: m is 1 or 3; Y is an alkylene group containing from 4 to 18 carbon 60 n is 1 or 2; atoms and separating the two 4-(R-NH)-1- x is 1, 2 or 3; and pyridinyl groups by from 4 to 18 carbon atoms; wherein R is an alkyl group containing from 6 to 18 carbon (m)(2)=(n)(x). atoms; 11. A compound according to claim 10 wherein R1 is R1 is hydrogen or lower alkyl; 65 hydrogen, Y is (CH2) and w is from 4 to 18. A is an anion; 12. A compound according to claim 11 wherein w is m is 1 or 3; from 8 to 12 and R is p-chlorophenyl. n is 1 or 2; 13. A compound having the formula 4,206,215 44 27. A skin-cleansing composition comprising an an

timicrobially effective amount of a compound accord ing to claim 5 wherein A is a pharmaceutically accept able anion, a compatible pharmaceutically acceptable 5 surfactant and a compatible, pharmaceutically accept able carrier. 28. An oral hygiene composition for the prevention of dental plaque comprising an effective amount of a wherein: compound according to claim 21 wherein A is a phar Y is an alkylene group containing from 4 to 18 carbon 10 maceutically acceptable anion and a compatible, phar atoms and separating the two 4-(R-NH)-1-pyridi maceutically acceptable carrier. nyl groups by from 4 to 18 carbon atoms; R is benzyl; 29. An antimicrobial composition suitable for applica R1 is hydrogen or lower alkyl; tion to inanimate surfaces comprising an effective 15 amount of a compound according to claim 1 in admix A is an anion; ture with a compatible vehicle. m is 1 or 3; n is 1 or 2; 30. A method for controlling microorganisms which x is 1, 2 or 3; and comprises contacting said microorganisms with an an wherein timicrobially effective amount of a compound of the (m)(2)=(n)(x). 20 formula 14. A compound according to claim 13 wherein R1 is hydrogen, Y is (CH2) and w is from 8 to 12. 15. 1,12-Bis-(4-(heptylamino)-1-pyridiniumdodecane W 7 --2 dibromide according to claim 6. RNH / Non-Y-N \ NHR 16. 1,12-Bis-(4-(heptylamino)-1-pyridinium)dodecane 25 An dichloride according to claim 6. 17. 1,10-Bis-(4-(octylamino)-1-pyridinium decane di R R1 n bromide according to claim 6. 18. 1,10-Bis-4-(octylamino)-1-pyridinium decane di wherein: chloride according to claim 6. 30 Y is an alkylene group containing from 4 to 18 carbon 19. 1, 12-Bis-4-(2-ethylhexylamino)-1-pyridinium atoms and separating the two 4-(R-NH)-1-pyridi dodecane dibromide according to claim 6. nyl groups by from 4 to 18 carbon atoms; 20. 1,12-Bis-4-(2-ethylhexylamino)-1-pyridiniumdo R is hydrogen, an alkyl group containing from 1 to 18 decane dichloride according to claim 6. carbon atoms, a cycloalkyl group containing from 21. A compound selected from the group consisting 35 5 to 7 carbon atoms, benzyl, or phenyl substituted of: by methylenedioxy or one or two substituents se 1.9-bis-(4-(heptylamino)-1-pyridiniumnonane dibro lected from the group consisting of halogen, lower mide, alkyl, lower-alkoxy, nitro, cyano and trifluoro 1,10-bis-(4-(2-ethylhexylamino)-1-pyridinium-dec methyl; ane dibromide, R1 is hydrogen or lower alkyl; 1,10-bis-(4-(octylamino)-1-pyridinium)decane dichlo A is an anion; ride, m is 1 or 3; 1.9-bis-(4-(octylamino)-1-pyridinium nonane dibro n is 1 or 2; mide, 45 x is 1, 2 or 3; and 1,12-bis-4-(heptylamino)-1-pyridiniumdodecane di wherein chloride, (m)(2)=(n)(x). 1,8-bis-(4-(octylamino)-1-pyridinium]octane dibro 31. A method according to claim 30 wherein R is mide and hydrogen. 1,10-bis-(4-(octylamino)-1-pyridinium decane dibro 50 mide. 32. A method according to claim 30 wherein R is an 22. 1.9-Bis-4-(heptylamino)-1-pyridinium nonane alkyl group containing from 1 to 18 carbon atoms, a dibromide according to claim 21. cycloalkyl group containing from 5 to 7 carbon atoms, 23. 1, 10-Bis-4-(2-ethylhexylamino)-1-pyridinium benzyl, or phenyl substituted by methylenedioxy or one decane dibromide according to claim 21. 55 or two substituents selected from the group consisting 24. 1, 10-Bis-4-(propylamino)-1-pyridinium decane of halogen, lower-alkyl, lower-alkoxy, nitro, cyano and dibromide. trifluoromethyl. 25. An antimicrobial composition suitable for topical 33. A method for controlling microorganisms which administration comprising an effective amount of a comprises contacting said microorganisms with an an compound according to claim 1 wherein A is a pharma timicrobially effective amount of a compound accord ceutically acceptable anion and a compatible, pharma ing to claim 3. " ceutically acceptable carrier. 34. A method for controlling microorganisms which 26. A skin-cleansing composition comprising an an comprises contacting said microorganisms with an an timicrobially effective amount of a compound accord timicrobially effective amount of a compound accord ing to claim 1 wherein A is a pharmaceutically accept 65 ing to claim 5. y able anion, a compatible, pharmaceutically acceptable 35. A method according to claim 34 wherein the surfactant and a compatible, pharmaceutically accept antimicrobially effective agent is 1,12-bis-(4-(hep able carrier...... ; tylamino)-1-pyridinium)dodecane dibromide. 4,206,215 45 46 36. A method according to claim 34 wherein the 41. A method of treating teeth for preventing the antimicrobially effective agent is 1,12-bis-(4-(hep formation of dental plaque thereon which comprises contacting the teeth with an effective amount of a com tylamino)-1-pyridiniumdodecane dichloride, pound according to claim 21. 37. A method according to claim 34 wherein the 42. A method according to claim 41 wherein the antimicrobially effective agent is 1,10-bis-(4-(oc dental plaque-preventive agent is 1.9-bis-(4-(hep tylamino)-1-pyridinium)decane dibromide. tylamino)-1-pyridiniumnonane dibromide, 38. A method according to claim 34 wherein the 43. A method according to claim 41 wherein the antimicrobially effective agent is 1,10-bis-(4-(oc dental plaque-preventive agent is 1,10-bis-(4-(2-ethyl tylamino)-1-pyridinium)decane dichloride, 10 hexylamino)-1-pyridinium decane dibromide. 44. A method according to claim 41 wherein the 39. A method according to claim 34 wherein the dental plaque-preventive agent is 1,10-bis-(4-(oc antimicrobially effective agent is 1,12-bis-(4-(2-ethyl tylamino)-1-pyridiniumdecane dichloride. hexylamino)-1-pyridiniumdodecane dibromide, 45. A method according to claim 41 wherein the 40. A method according to claim 34 wherein the dental plaque-preventive agent ... is 1.9-bis-4-(oc antimicrobially effective agent is 1,12-bis-(4-(2-ethyl tylamino)-1-pyridiniumnonane dibromide, hexylamino)-lpyridiniumdodecane dichloride,

25

35

45

55

65