JPS6225153B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a new penicillin, and more specifically, it has the general formula (1). The present invention relates to penicillanic acid represented by the formula (wherein, Y represents a hydrogen atom or a hydroxyl group), physiologically acceptable salts and esters thereof, and antibacterial agents containing these as active ingredients. The penicillin of the present invention has a broad antibacterial spectrum and has strong antibacterial properties against Gram-negative bacteria, particularly Pseudomonas and Escherichia coli. To give a more detailed explanation of the penicillin of the present invention, penicillanic acid represented by the general formula () is 6-[D-(-)-α-(4-hydroxy-5,6,7,8- Tetrahydro-5-oxo-3-quinolinecarboxyamido)-phenylacetamide] penicillanic acid and 6-[D-
(-)-α-(4-hydroxy-5, 6, 7, 8-
Tetrahydro-5-oxo-3-quinolinecarboxamide)-α-(4-hydroxyphenyl)acetamide] penicillanic acid can be mentioned. Physiologically acceptable salts and esters of these penicillanic acids are also included within the scope of the penicillin of the present invention. The physiologically acceptable salts mentioned above include alkali metal salts such as lithium salts, sodium salts, potassium salts, etc.; alkaline earth metal salts such as calcium salts, magnesium salts, etc.; lysine salts, arginine salts, ornithine salts. Basic amino acid salts such as salts; common organic base salts such as triethylamine salts, benzylamine salts, procaine salts, etc. can be mentioned. In addition, examples of the above-mentioned physiologically acceptable esters include penicillanic acid and lower alcohol represented by the general formula (), and ordinary esters containing an ester group represented by the following formula. -CHR ₁ -X-CO-R ₂ (In the formula, R ₁ represents a hydrogen atom or a lower alkyl group, and R ² represents a lower alkyl group. X represents an oxygen atom or a sulfur atom.) Penicillin of the present invention is the general formula (2) (wherein, Y represents the same meaning as in general formula (1)), a salt or ester thereof, and formula (3) 4-hydroxy-5,6,7,8- represented by
It can be produced by reacting a reactive derivative of tetrahydro-5-oxo-3-quinolinecarboxylic acid. In addition, 6-aminopenicillanic acid, its salt or ester has the general formula (4). (In the formula, Y represents the same meaning as in general formula (1).) It can also be produced by reacting a reactive derivative of a carboxylic acid represented by the following. The compound represented by general formula (2) is a compound known as ampicillin or amoxicillin.
The quinoline carboxylic acid represented by formula (3) is a new compound and can be produced by the method shown in the following formula and the reference examples below.

【table】

[Table] Examples of the reactive derivative of quinoline carboxylic acid represented by formula (3) include acid halides and mixed acid anhydrides. The reaction of the compound represented by general formula (2), its salt or ester with the reactive derivative of quinoline carboxylic acid represented by formula (3) is usually carried out in a reaction solvent. As a reaction solvent, tetrahydrofuran, methylene chloride, chloroform, dioxane,
Acetate ester and the like are suitable. The reaction between 6-aminopenicillanic acid, its salt or ester and the reactive derivative of the carboxylic acid represented by the general formula (4) can be carried out in an aqueous solvent or a non-aqueous organic solvent. The carboxylic acid represented by general formula (4) can be synthesized by reacting phenylglycine or 4-hydroxyphenylglycine with a reactive derivative of quinoline carboxylic acid represented by formula (3). As a reactive derivative of the carboxylic acid represented by the general formula (4), the formula
As in the case of the reactive derivative of quinoline carboxylic acid represented by (3), acid halides, mixed acid anhydrides, etc. are suitable. Hereinafter, the method for producing penicillin of the present invention will be specifically explained using reference examples and synthesis examples. Reference Example 1 Synthesis of 4-hydroxy-5,6,7,8-tetrahydro-5-oxo-3-quinolinecarboxylic acid (1) N-(3-oxo-1-cyclohexene-1
Synthesis of diethyl-yl)-aminomethylene malonate 11.1 g of 3-amino-2-cyclohexenone,
24.2 g of diethyl ethoxymethylenemalonate and 0.12 g of p-toluenesulfonic acid were mixed and reacted on an oil bath at 120-130°C for 2 hours. The product was purified by column chromatography and N
-(3-oxo-1-cyclohexen-1-yl)-aminomethylene diethyl malonate 19.8g
was obtained as a pale yellow viscous oil. Nuclear magnetic resonance spectrum ( _d6 -DMSO, 60Mc,
TMS) is δ: 1.24 (3H, t, J = 7Hz),
1.26 (3H, t, J=7Hz), 1.7~2.75 (6H,
m), 4.10 (2H, q, J=7Hz), 4.18 (2H,
q, J=7Hz) 5.75 (H, s), 8.05 (1H,
d, J = 13.8Hz) and 10.2 (1H, d, J =
It had a signal at 13.8Hz). (2) Synthesis of ethyl 4-hydroxy-5,6,7,8-tetrahydro-5-oxo-3-quinolinecarboxylate Heat 20 ml of diphenyl ether to 260°C or higher, add N-(3-oxo- 6.87 g of diethyl 1-cyclohexen-1-yl)-aminomethylenemalonate was dissolved in 5 ml of diphenyl ether and added dropwise, followed by refluxing for 15 minutes. After cooling,
The reaction mixture was poured into 300 ml of n-hexane, and the precipitate was collected. This precipitate was purified to obtain 4.37 g of a pale yellow powder, which was the desired product. The melting point of this product is 221-224℃ (decomposition), and its infrared absorption spectrum (KBr tablet) is 3340,
2960, 1740, 1690, 1635, 1605, 1560, 1510,
1285, 1170, 1150, 1090, 1035, 920 and 815
It had maximum absorption at cm ^-1 . Also,
The nuclear magnetic resonance spectrum (d ₆ −
DMSO, 60Mc, TMS), δ: 1.31 (3H,
t, J=7Hz), 1.9-3.1 (6H, m), 4.27
(2H, q, J=7Hz) and had signals at 8.55 (1H, s). (3) Synthesis of 4-hydroxy-5,6,7,8-tetrahydro-5-oxo-3-quinolinecarboxylic acid 4-hydroxy-5,6,7,8-
4.0 g of ethyl tetrahydro-5-oxo-3-quinolinecarboxylate and 2.9 g of caustic soda were added, and the mixture was stirred at 90 to 95°C for 2 hours. cooling,
The reaction solution was adjusted to pH=2 with 6H hydrochloric acid, and the precipitated crystals were collected and washed with water and ethanol.
Dry under reduced pressure at 120°C for 4 hours to obtain the target compound.
3.3g was obtained. The melting point of this substance is 278-279℃
(decomposition), and its infrared absorption spectrum (KBr tablet) is 3420, 1740, 1690,
It had maximum absorption at 1640, 1500 and 820 cm ^-1 . Synthesis example 1 6-[D-(-)-α-(4-hydroxy-5.
6,7,8-tetrahydro-5-oxo-3-
Synthesis of penicillanic acid (quinolinecarboxamide)-phenylacetamide 4-hydroxy-5, 6,
1.13 g of 7,8-tetrahydro-5-oxo-3-quinolinecarboxylic acid and 1.1 g of triethylamine
After cooling to -15°C, 1.18 g of ethyl chloroformate dissolved in 5 ml of methylene chloride was added dropwise. After stirring at -15°C for 30 minutes, ampicillin trihydrate was mixed with 20 ml of methylene chloride and 5 ml of dioxane.
ml of triethylamine and 1.0 g of triethylamine. After reacting at -15 to 0°C for 1.5 hours, the solvent was distilled off, and 1.37 g of potassium carbonate dissolved in 90 ml of water was added to the residue, followed by stirring at room temperature for 1 hour.
After the reaction, the pH of the reaction solution was adjusted to 2 with 40% phosphoric acid, the solution was saturated with common salt, and the precipitated crystals were collected.
Next, this was purified using a silica gel packed column to obtain 0.85 g of the target compound. Infrared absorption spectrum of this product (KBr tablet)
are 3060, 2960, 1795, 1760, 1690, 1620,
It has maximum absorption at 1505, 1410, 1340, 1300, 1200, 1090, 1040 and 810 cm ^-1 , and the nuclear magnetic resonance spectrum ( _d6 -DMSO, 60Mc,
TMS) is δ: 1.42 (3H, s), 1.57 (3H,
s), 1.75-3.1 (6H, m), 4.20 (1H, s), 5.25
~5.70 (2H, m), 5.91 (1H, d, J=8Hz),
7.1-7.65 (5H, m), 8.35 (1H, s), 9.17
(1H, d, J = 8Hz) and 10.99 (1H, d, J
= 8 Hz). Synthesis example 2 6-[D-(-)-α-(4-hydroxy-5.
6,7,8-tetrahydro-5-oxo-3-
Synthesis of penicillanic acid (quinolinecarboxamide)-α-(4-hydroxyphenyl)acetamide] 4-hydroxy-5, 6,
1.14 g of 7,8-tetrahydro-5-oxo-3-quinolinecarboxylic acid and 1.1 g of triethylamine
After cooling to -15°C, 1.20 g of ethyl chloroformate dissolved in 5 ml of methylene chloride was added dropwise. After stirring at -15℃ for 30 minutes, amoxicillin・
2.10 g of the trihydrate was dissolved in a mixture of 10 ml of methylene chloride, 10 ml of dimethylformamide and 1.0 g of triethylamine and added. After reacting at -10 to 0°C for 1.5 hours, the reaction solution was poured into ether petroleum benzine mixture 1. The precipitate was collected and purified using a silica gel packed column. Next, it was dissolved in a small amount of water, the pH was adjusted to 2 with 40% phosphoric acid, the precipitated crystals were collected, and the target compound was 0.28
g was obtained as a white powder. The infrared absorption spectrum (KBr tablet) of this product is 3400-3240, 1785,
It has maximum absorption at 1760, 1685 and 1505 cm ^-1 , and the nuclear magnetic resonance spectrum (d ₆ -
DMSO, 60Mc, TMS), δ: 1.40 (3H,
s), 1.54 (3H, s), 1.75-3.1 (6H, m), 4.08
(1H, s), 5.3-5.8 (3H, m), 6.63 (2H, d,
J=8Hz), 7.16 (2H, d, J=8Hz), 8.27
(1H, s), 9.02 (1H, d, J=8Hz) and
It had a signal at 10.85 (1H, d, J=8Hz). The antibacterial action of the penicillin of the present invention can be demonstrated by the following compound A.
and the results measured for Compound B are described below. Comparative control drugs include the following known compounds, aminobenzylpenicillin sodium (compound AB-PC), carbenicillin sodium (compound CB-PC), and 6-[D-(-)-α-(4-hydroxy-1.5 -naphthyridine-3-carboxyamide)-phenylacetamide] Sodium penicillanate (compound C) was used. Compound A Compound B Compound AB-PC Compound CB-PC Compound C Antibacterial activity against Gram-positive and Gram-negative bacteria is determined by the minimum inhibitory concentration method (MIC) using conventional methods.
method). The experimental procedure was as follows. As a measurement medium and an enrichment medium for test bacterial strains,
Commercially available heart infusion agar medium (abbreviated as HIA medium) and trypto soy broth medium (abbreviated as TSB medium) were used, and the test compound was serially diluted with molten HIA medium and dispensed into sterile petals. A drug plate was prepared by cooling and solidifying it. The test bacteria were enriched in TSB medium at 37℃ for 18 hours, and then the culture solution was grown in freshly prepared TSB medium for 50 to 50 minutes.
The mixture was diluted 500 times and inoculated one platinum loop onto the drug plate described above. Next, the drug plate was incubated at 37°C for 18 hours, and the growth status of bacteria on the plate was examined to determine the minimum inhibitory concentration (MIC). The results are in the table-
As shown in 1. As is clear from Table 1, the compound of the present invention has a broad antibacterial spectrum and has a high level of antibacterial activity, especially against Pseudomonas aeruginosa compared to compound CB-PC and other control drugs. It can be said that it is clearly superior in comparison. Next, experimental examples will be shown below regarding acute toxicity in mice, blood concentration and tissue concentration in mice, and serum protein binding rate of Compound A and Compound B of the present invention. Experimental Example 1 Regarding the above-mentioned compound A and compound B of the present invention,
The 50% lethal dose (abbreviated as LD ₅₀ value) was estimated by intravenous administration using 10 male and female ICR mice each with a volume of 25 g. Both doses of compound A and compound B are 500 mg/Kg,
Two groups of 1000 mg/Kg were administered to mice by the usual method, followed by follow-up observation for 2 weeks after administration into the tail vein. As a result, the LD ₅₀ values of Compound A and Compound B were estimated to be 1000 mg/Kg or more in both males and females. Experimental Example 2 The blood and tissue concentrations of Compound A of the present invention when subcutaneously administered to mice were measured in comparison with Compound C, a control drug. Each group of 5 ICR male mice weighing around 25 g was used, and both drugs were injected subcutaneously into the mice at 50 mg/Kg using a conventional method. 15 minutes, 30 minutes, 60 minutes after injection
Blood was collected by cutting the carotid arteries and veins, and pooled serum from one group was used for blood concentration measurement. In addition, for both drug administration groups 15 minutes after injection, the heart, lungs, liver, and spleen were removed, homogenates of these organs were prepared by grinding in a mortar, and the centrifuged supernatants were used to measure tissue concentrations. The blood and tissue concentrations of both drugs are
The test was carried out by the thin layer cup method using Micrococcus luteus ATCC 9341 strain as the test bacterium. The measurement results of blood concentration and tissue concentration are shown in Table 2 and Table 3, respectively. The blood and tissue concentrations of Compound A of the present invention when administered subcutaneously to mice were always as high as those of Compound C, a control drug. Experimental Example 3 The blood concentration of Compound B of the present invention when subcutaneously administered to mice was compared with that of Compound C, a control drug compound CB-PC, and Compound C. One ICR male mouse weighing around 35 g
A group of 5 mice were used, and each of the three drugs was injected subcutaneously at a dose of 50 mg/Kg using a conventional method. Example 2 above
Blood was collected at 15, 30, and 60 minutes after injection in the same manner as above, and the pooled serum of Group 1 was used for blood concentration measurement. Blood concentration is
Bioassay was carried out by the thin layer cup method using Micrococcus luteus ATCC 9341 strain as the test bacterium. The measurement results are shown in Table 4, and the blood concentration of Compound B of the present invention was superior to Compound C and almost equal to Compound CB-PC. Experimental Example 4 The rat serum protein binding rate of Compound A of the present invention was determined by the method of Gomi et al. (Chemotherapy, 17, p. 1363, 1969).
Comparative measurements were made with the control drug Compound C using an equilibrium dialysis method according to . Pour 4 ml of 80% rat serum containing drugs as an internal solution into a cellulose tube, add 26 ml of M/15 phosphate buffer solution (PH7.0) as an external solution, and
Equilibrium dialysis was performed at ℃ for 48 hours, and the drug concentration in the internal and external fluids was calculated by bioassay using the thin-layer cup method using Micrococcus luteus ATCC 9341 strain as the test bacterium. The results are shown in Table 5, and the rat serum protein binding rate of the compound A of the present invention was lower than that of the control drug compound C.

【table】

【table】

【table】

【table】

【table】

[Table] As is clear from the above description, the penicillin of the present invention has extremely excellent antibacterial activity and is useful as an antibacterial agent, and is used for the treatment and prevention of bacterial infections in mammals including humans. Not only can it be used, but it can also be used as a disinfectant. When administered to humans, the single dose is 100 mg to 1500 mg, preferably 250 mg to 1000 mg, and it is desirable to administer it several times a day. The dosage form of the preparation containing the compound of the present invention as an active ingredient may be a solid preparation such as a tablet, capsule, or powder, or a liquid preparation such as an injection solution or suspension.
As excipients, stabilizers, wetting agents, etc., those commonly used in the field may be used.

Claims (1)

[Claims] 1 General formula () (In the formula, Y represents a hydrogen atom or a hydroxyl group.) Penicillanic acid, physiologically acceptable salts and esters thereof. 2. The chemical substance according to claim 1, wherein Y in the general formula () is a hydrogen atom. 3. The chemical substance according to claim 1, wherein Y in the general formula () is a hydroxyl group. 4 General formula () An antibacterial agent containing penicillanic acid represented by the formula (wherein Y represents a hydrogen atom or a hydroxyl group), a physiologically acceptable salt or ester thereof, as an active ingredient. 5. The antibacterial agent according to claim 4, wherein Y in the general formula () is a hydrogen atom. 6. The antibacterial agent according to claim 4, wherein Y in the general formula () is a hydroxyl group.