Patent GB-1107790-A (GB)
Title: A process and diagnostic agents for the detection of bacteria
IPC: C07D333/44
Filed: 1967-03-23
Abstract: Compounds of the following general formula are useful for the detection of bacteria, especially in urine, milk and drinking water, by a colour reaction:- <FORM:1107790/C6-C7/1> wherein X is an oxygen or sulphur atom or an alkylated nitrogen atom and R is one of the following radicals a, b, c, d or e: <FORM:1107790/C6-C7/2> <FORM:1107790/C6-C7/3> <FORM:1107790/C6-C7/4> <FORM:1107790/C6-C7/5> <FORM:1107790/C6-C7/6> wherein R1 is a saturated or unsaturated aliphatic, aromatic, araliphatic acyl, hydroxy or acyloxy aliphatic radical, R2 is a hydrogen atom or a lower alkyl radical, R3 and R4 are both hydrogen atoms or together form an isopropylidene radical, and n is 2 or 3.  The procedure for the preparation of the following compounds or classes of compounds, is described: (1)N-substituted 2,4-di-(5-nitro-2-furfurylidene)-granatan-3-one and 2,4-di-(5-nitro-2-furfurylidene)-tropinone derivatives of formula 1a, by heating the N-substituted norgranatan -3-one or tropinone in question with 5-nitrofurfural to give compounds wherein X is oxygen and n is 3 (except where stated otherwise), and R1 is -CH2.CH2.OCOCH3, -CH3 (n is 2 or 3), -C2H5, -C6H5, -CH2.C6H5, -CH2.CH=CH2, -CH2.CH2.C6H5, -CO.CH3, -CH3 (X is sulphur), and OH; the preparatio of N-hydroxy-norpseudopelletierin, required as starting material for the preparation of the last compound, from acetone dicarboxylic acid, sodium acetate trihydrate, hydroxylamine hydrochloride and glutaric dialdehyde solution, is also given; (2) 5-nitro-furfurylidene derivatives of homophthalimide of formula 1b from the monophthalimide and 5-nitro-furfural; (3) 3-(5-nitro-2-thienyl)-2-nitro-propane-1, 3-diol and 2, 2-dimethyl-4-(5-nitro-2-thienyl)-5-nitro-1, 3-dioxan of formula 1c from 5-nitro-thiophene-2-aldehyde, nitro-ethanol and triethylamine; (4) 1-(5-nitro-1-methyl-2-pyrryl)-2-(4-quinolyl)-ethylene of formula 1a from 4-methyl-quinoline and 5-nitro-1-methyl-pyrrole-2-aldehyde; and (5) 1-phenyl-3-methyl-4-(5-nitro-2-furfurylidene)-pyrazol-5-one of formula 1e from 1-phenyl-3-methyl-pyrazol-5-one and 5-nitrofurfural.  The material to be investigated is brought together with a solid or liquid nutrient medium (specified) and an amount of the diagnostic compound insufficient to inhibit any microbial growth, and the colour change is evaluated after incubating for 1-18 hours at 20-37 DEG C. Preferably the diagnostic compound is added after an initial incubation has taken place without it.  There are preferably included for the diagnostic test: (1) at least one co-enzyme (nicotinamide-adenine-dinucleotide, nicotinamideadenine-dinucleotide phosphate or a reduction product thereof); (2) a sulphydryl group - containing compound or a corresponding dehydrated disulphide compound (cysteine, reduced glutathione, cysteamine or b -mercapto-ethanol); and/or (3) enzyme-activating heavy metal ions (manganese or copper). The components may be put up in the form of a tablet or impregnated filter paper, which may be sterilized with ethylene oxide and sealed for storage between sterilized foils.  Alternatively, the diagnostic compound may be in the form of a stable solution, containing a solubilizer (dimethyl formamide and polyoxyethylene sorbitan laurate) if necessary, with the other components formulated in a gelatine capsule from which they can be removed and dissolved in water prior to use. Examples are directed to the detection by colour reaction of Escherichia coli, Streptococcus faecalis, Proteus mirabilis, Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa.  In certain cases, semi-quantitative determinations are made based upon the time to achieve an anticipated change or graduation in colour.ALSO:Compounds of the following general formula are useful for the detection of bacteria, especially in urine, milk and drinking water, by a colour reaction:-  <FORM:1107790/C2/1> wherein X is an oxygen or sulphur atom or an alkylated nitrogen atom and R is one of the following radicals a, b, c, d, or e:- <FORM:1107790/C2/2> <FORM:1107790/C2/3> <FORM:1107790/C2/4> <FORM:1107790/C2/5> <FORM:1107790/C2/6> wherein R1 is a saturated or unsaturated aliphatic, aromatic, araliphatic, acyl, hydroxy or acyloxy aliphatic radical, R2 is a hydrogen atom or a lower alkyl radical, R3 and R4 are both hydrogen atoms or together form an isopropylidene radical, and n is 2 or 3.  The procedure for the preparation of the following compounds or classes of compounds, is described: (1) N-substituted 2,4 - di - (5 - nitro - 2 - furfurylidene) - granatan-3-one and 2,4-di-(5-nitro-2-furfurylidene)-tropinone derivatives of Formula 1 (A), by heating the N-substituted norgranatan-3-one or tropinone in question with 5-nitrofurfural to give compounds wherein X is oxygen and n is 3 (except where stated otherwise), and R1 is -CH2.CH2.OCOCH3, -CH3 (n is 2 or 3), -C2H5, -C6H5, -CH2.C6H5, -CH2.CH = CH2, -CH2.CH2.C6H5, -CO.CH3, -CH3\t (X is sulphur), and -OH; the preparation of N-hydroxy-norpseudopelletierin, required as starting material for the preparation of this last compound, from acetone dicarboxylic acid, sodium acetate trihydrate, hydroxylamine hydrochloride and glutaric dialdehyde solution, is also given; (2) 5-nitro-furfurylidene derivatives of homophthalimide of Formula 1 (B) from the monophthalimide and 5-nitro-furfural; (3) 3-(5-nitro - 2 - thienyl - 2 - nitro - propane - 1,3 - diol and 2,2 - dimethyl - 4 - (5 - nitro - 2 - thienyl) -5-nitro-1,3-dioxan of Formula 1(C) from 5-nitro-thiophene-2-aldehyde, nitro-ethanol and triethylamine; (4) 1-(5-nitro-1-methyl-2-pyrryl)-2-(4-quinolyl)-ethylene of Formula 1 (D) from 4 - methyl - quinoline and 5 - nitro - 1 - methyl - pyrrole - 2 - aldehyde; and (5) 1 - phenyl - 3 - methyl - 4 - (5 - nitro - 2 - furfurylidene) - pyrazol - 5-one of Formula 1 (E) from 1-phenyl-3-methyl-pyrazol-5-one and 5-nitrofurfural.