JPS5919946B2 - 2-aminothiazol-4-ylglycine derivative - Google Patents

Description

Detailed Description of the Invention The present invention is based on the general formula R2NH Page/COORI CHCI] [In the formula, -COORI represents an optionally protected carboxyl group, and R2NH- and R3NH- represent an optionally protected amino group. shows.

] This relates to a 2-aminothiazole-4-ylglycine derivative represented by the following. As a result of various studies, the present inventors found that the general formula R2NH□/COORI C [■] [wherein -COORI and R2NH- have the same meanings as above.

] by reducing the 2-aminothiazole derivative represented by
Protecting groups can be removed if necessary, or general formula XC
H2-C-CH-COORI I)l [■] [In the formula, X is a halogen atom, -COORI and R3
NH- has the same meaning as above.

[In the formula, R2NH- has the same meaning as above.

2-aminothiazol-4-ylglycine derivative [1] can be obtained by reacting with a thiourea compound represented by It has been discovered that penicillins or cephalosporins in which the amino group at the 7-position is acylated have excellent antibacterial activity, and the present invention has been completed based on this finding. In the above formula, -COORl represents an optionally protected carboxyl group, and therefore R1 represents a hydrogen atom or a protecting group for the carboxyl group.

Any type of protecting group may be used as long as it can be removed under mild conditions such as acidic conditions, alkaline conditions, reducing conditions, etc. without affecting the thiazole nucleus; for example, it is generally used as a protecting group for carboxyl groups in peptide synthesis. For example, alkyl groups such as methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, substituted alkyl groups such as β-methylsulfonylethyl, trichloroethyl, diphenylmethyl, phenyl, Aryl groups such as tolyl, substituted aryl groups such as p-tert-butylphenyl, p-nitrophenyl, aralkyl groups such as benzyl, phenethyl, tolbenzyl, p-methoxybenzyl, p-
Substituted aralkyl groups such as nitrobenzyl are used. R2NH- and R3NH- represent an optionally protected amino group, and therefore R2 and R3 represent a hydrogen atom or a protecting group for the amino group. As such protecting groups, protecting groups for amino groups that are easily removed and are commonly used in peptide chemistry are used, such as alkylcarbonyl groups such as formyl, acetyl, and propionyl, alkoxycarbonyl groups such as t-butoxycarbonyl, and methoxycarbonyl groups. Alkoxyalkylcarbonyl groups such as acetyl and methoxypropionyl, substituted alkoxycarbonyl groups such as trichloroethoxycarbonyl, alkoxycarbonylacetyl groups such as methoxycarbonylacetyl, aralkyloxycarbonyl groups such as benzyloxycarbonyl, and substitutions such as p-nitrobenzyloxycarbonyl. Aralkyloxycarbonyl groups are often used. X represents a halogen atom such as chlorine, bromine, iodine, or fluorine.
In the method for producing the 2-aminothiazole-4-ylglycine derivative [1] of the present invention, the 2-aminothiazole derivative [] is first reduced.

For the reduction method, a known reduction method that can lead to the target product [1] is used, such as using a catalyst (for example, Raney-nickel, platinum oxide, palladium carbon, ruthenium carbon, rhodium carbon, copper chromium oxide, etc.) Catalytic reduction method, reduction means using hydrogen generated by the combination of metals such as sodium, sodium amalgam, and aluminum amalgam with water, alcohol, etc., lithium aluminum hydride, diethyl aluminum hydride, sodium aluminum hydride, Reduction means using a complex metal hydride such as sodium borohydride, treatment with a metal such as zinc or iron in acetic anhydride, formic acid or a mixed solvent of water, electrolytic reduction, etc. are conveniently used. The temperature, pressure, type of solvent, time, and other conditions for the reduction reaction are appropriately determined depending on the type of raw material and the reduction means. After completion of the reduction, the protecting group can be removed as necessary, with or without separating the target product [1]. The protective group can be removed using the means normally used for removing each protective group. For example, an alkyl group such as ethyl or propyl can be removed using an acid or alkali, a diarylalkyl group such as diphenylmethyl, or a diarylalkyl group such as p-methoxybenzyl can be removed. Alkoxyaralkyl groups are treated with acids, methylsulfonylalkyl groups such as β-methylsulfonylethyl are treated with alkalis, and halogenoalkyl groups such as β, β, β-trichloroethyl are treated with p-nitrochloride and acid. p-nitroaralkyl groups such as benzyl can be easily reduced by catalytic reduction. The protective groups R2 and R3 of the amino group can be removed, for example, by removing a halogenoalkoxycarbonyl group such as β, β, and β-trichloroethoxycarbonyl with zinc and an acid;
A substituted aralkyloxycarbonyl group such as p-nitrobenzyloxycarbonyl can be easily prepared by catalytic reduction. When both the carboxyl group and the amino group are protected, these protecting groups may be applied at the same time, or one of them may be removed first, taking into account the type of protecting group. The obtained target product [1] can be obtained by known means such as solvent extraction, dissolution, liquid conversion, crystallization, recrystallization, distillation,
It can be isolated and purified by chromatography, ion exchange, etc. The isolated target product [1] is a DL mixture containing an appropriate optically active crystalline salt such as tartaric acid,
It is possible to split it into D-form and L-form by converting it into mandelic acid, malic acid, gamphasulfonate, etc. Moreover, the object compound of the present invention [] is α-amino-β
It can be produced by reacting -keto-γ-halogenobutyric acid esters [] with a thiourea compound [] and removing the protecting group as necessary.

In the reaction, it is preferable to react approximately equivalent amounts of the raw material compounds [] and []. The reaction is usually carried out in a solvent,
As such a solvent, an organic solvent that does not interfere with the reaction is used. For example, methanol, ethanol, propanol, tetrahydrofuran, etc. are frequently used. A base may be added to such solvents to advantageously advance the reaction. As the base, for example, pyridine, triethylamine, N-N-dimethylaniline, etc. are used. These bases are 100~
Add 300%, preferably 100-200%. When R2 is a hydrogen atom, it is not necessary to add a base because [] itself acts as a base. The reaction proceeds suitably at room temperature to reflux conditions. Usually, the reaction is completed in several tens of minutes to several hours. After the reaction is completed, the protective group is removed as needed by using the reaction solution as it is or by separating [1].
As the removal means, the same means as described above can be used.
The target product [1] can be separated by the same isolation and purification means as in the above method. The raw material compound [ ] of the present invention has the general formula [wherein, X and -COO
Rl has the same meaning as above.

It can be produced by reacting α-oximino-β-keto-γ-halogenobutyric acid esters represented by ] with a thiourea compound [ ] and F.

This reaction is carried out in an organic solvent such as ethanol or tetrahydrofuran. A base is preferably added to the solvent, and examples of such bases include tertiary bases such as pyridine, triethylamine, and N.N-dimethylaniline. When R2 is a hydrogen atom, [] itself acts as a base, so there is no need to further add a base. The reaction advantageously proceeds at room temperature to reflux conditions. The obtained raw material compound 2-aminothiazole derivative [] can be isolated and purified by known means, but it can also be used as a raw material for the method of the present invention as a reaction solution without separation. The object compound [1] of the present application obtained in this way is a novel intermediate useful for the production of penicillins or cephalosporins having a 2-aminothiazol-4-ylglycylamide group at the 6th or 7th position and having excellent antibacterial activity. be. Example 1 (1) 19.37 ethyl α-oximino-β-oxo-r-chlorobutyrate and 8.07 ml of urea were mixed with 20 ml of ethanol.
Dissolve in 0mj and heat under reflux for 2 hours.

Ethanol was distilled off under reduced pressure, and the residue was dissolved in 10% hydrochloric acid. In order to remove unreacted ethyl α-oximino-β-oxo-r-chlorobutyrate from this solution, it was washed twice with 200 ml each of ether, and then sodium hydrogen carbonate was added to the solution to pH 7. Adjust the temperature to 0 to 7.5 and extract twice with 100 ml each of chloroform. Wash the chloroform layer with water,
After drying, chloroform was distilled off to give 2-aminothiazole-
Ethyl 4-yl-α-oximinoacetate is obtained. Yield: 6.4370 This product has a melting point of 137-138°C (decomposition point). Elemental analysis value C7H9O3N3S Calculated value: C39. O6; H4.2l; Nl9.52 Actual value: C39.64: H4. O9;Nl9. O2 infrared absorption spectrum (NujOl) is 3430? -1 and 1710c of oxime! Absorption of ester is shown at n-1.

(2) Add 2.15 tons of ethyl 2-aminothiazol-4-yl-α-oximinoacetate obtained in (1) to 20% of 50% formic acid.
ml and 10 ml of methanol.

While stirring under ice-cooling, 1.5 y of zinc powder was added little by little to the mixture, stirring was continued for 3 hours, and then insoluble materials were removed. The solution is concentrated under reduced pressure to obtain a solution containing crude 2-aminothiazol-4-ylglycineethyl. Add this as it is to strong acidic ion exchange resin (H type) Amberlite IR-12.
The formic acid is removed by passing through a column consisting of 0.060 ml and repeated washing with water.

Thereafter, the eluate was eluted with 10% aqueous ammonia, and the eluate was concentrated under reduced pressure to obtain 2-aminothiazol-4-ylglycine. Yield 1.49t (yield 86.4%). This product is purified by recrystallization from aqueous ethanol and has a melting point of 186-190.
Indicates °C (decomposition point). Nuclear magnetic resonance spectrum (60Mc
, in trifluoroacetic acid) shows a single line of side chain methine hydrogen at 5.25 ppm and a single line resonance line of hydrogen on the thiazole ring at 6.75 ppm.

Gives a purple color with ninhydrin reagent. (3) Suspend 3.46 f of 2-aminothiazol-4-ylglycine in 100 m2 of N-N-dimethylacetamide.

While stirring at room temperature, 12.66 V of β·β·β monotrichloroethoxycarbonyl chloride was added dropwise over 30 minutes. After stirring for a further 30 minutes, 250 ml of ethyl acetate are added to the reaction mixture. After washing with 707 n1 of 1N hydrochloric acid, the ethyl acetate layer is separated.

This ethyl acetate layer was diluted with 50 ml of 3% caustic potash aqueous solution.
Extract times. After washing the aqueous layer once with ethyl acetate,
PH2. with normal hydrochloric acid. 100ml of ethyl acetate to precipitate the oily substance! Extract 3 times. Wash the ethyl acetate layer with water,
After drying with magnesium sulfate, the residue was distilled off under reduced pressure. Addition of n-hexane to the residual oil results in crystallization. Recrystallize from a mixture of ethyl acetate and ligroin to obtain 2-(β·β·β-trichloroethoxycarbonylamino)thiazol-4-yl-N-(β·β·β-trichloroethoxycarbonyl)glycine 510W1f. . Nuclear magnetic resonance spectrum of this product (100MHz in monochloroform)
is two overlapping single lines based on trichlorochlorethoxy group hydrogen at 4.80 ppm, a single line of 2-position amino group hydrogen at 4.65 ppm, a wide double line of methine hydrogen at 5.48 ppm, ε 1A simple M7 /7'd-Maz no T4i theory D
The single resonance line of hydrogen at the 5-position of thiazole is shown at 6.95 ppm.

Example 2 Ethyl 2-aminothiazol-4-yl-α-oximinoacetate hydrochloride 503W! f in 50% formic acid aqueous solution 10ml
and 5 ml of ethanol.

Add zinc powder 300rf9 little by little to this while stirring on ice. After adding the entire amount and stirring for another 3 hours, ethanol was distilled off under reduced pressure at 30°C or below, and 1N-NaOH was added to the residue.
Add to adjust the pH to 7.5. After extraction with ethyl acetate, the ethyl acetate layer was washed with water and dried, and the ethyl acetate was distilled off to give 2-
Aminothiazol-4-ylglycine ethyl ester is obtained in crystalline form. Yield: 130 WIf (32.3%).
This product becomes colored and decomposes when exposed to air. 1.04p in nuclear magnetic resonance spectrum (60Mc1 in trifluoroacetic acid)
Triple line of methyl hydrogen at pm, quartet line of methylene hydrogen at 4.18 ppm, single line of methylene hydrogen at 5.35 ppm,
A single resonance line of hydrogen at the 5-position of the thiazole ring is shown at 6.90 ppm. The mass spectrometry spectra shows a theoretical value of 201.0571.
The actual measured value was 201.0549. Example 3 26.6 V of ethyl α-acetamido-β-oxo-γ-bromobutyrate was added to 50 ml of ethanol and 20 ml of ether.
9.14 f7 of thiourea and 15 ml of pyridine were added thereto, and the mixture was stirred at room temperature for 1 hour and then under reflux for 4 hours.

After concentrating the reaction solution, 50a of ethyl acetate was added to the residue, and the mixture was extracted with 3N HCl. pH the aqueous layer with 1N-NaOH
Bring to 1O and extract with ethyl acetate. Wash the ethyl acetate layer with water,
After drying, the mixture is concentrated and chloroform is added to obtain ethyl 2-aminothiazol-4-yl-α-acetamidoacetate in the form of crystals. Yield 7.0y (34.5%). This product has a melting point of 161.1℃. Example 4 34.67 ethyl α-acetamido-β-oxo-γ-bromobutyrate was dissolved in ethanol 50
m1 and 20 m1 of ether.

To this were added 18.9 r of N-acetylthiourea and 15 ml of pyridine, and the mixture was heated under reflux for 4 hours. After distilling off the solvent under reduced pressure, 150 ml of ethyl acetate was added, and the ethyl acetate layer was washed with a 5% aqueous sodium bicarbonate solution, then water, and dried. The oil obtained by concentrating ethyl acetate was purified by passing it through a silica gel column to obtain 2-acetamidothiazole-4.
-yl-α-acetamidoethyl acetate is obtained. Yield 4.
467 (15.6%). This product has a melting point of 148.9-150
．． Example 5 at 1°C
1.87% of N-N-dimethylaniline was added thereto, and the mixture was stirred at room temperature for 24 hours.

After concentrating the reaction solution, 30 ml of chloroform was added to dissolve it, and the chloroform layer was washed with 3N-HCl, then water, and after drying, the chloroform was distilled off. The semi-solid obtained was purified by passing it through a silica gel column to obtain 2-(β・Ethyl .beta..beta.-trichloroethoxycarbonylamino)thiazol-4-yl-.alpha.-acetamidoacetate is obtained in crystalline form.
Yield 1.48f7 (35.3%). Melting point: 161.9°C.
Elemental analysis value Cl2Hl4N3O5SCl3・-f-H
2ON-acetylglycine is obtained as white crystals. Yield 65η (70%). Melting point: 158.0°C o Nuclear magnetic resonance spectrum (100 Mc, in deuterated dimethyl sulfoxide) shows a single line of acetyl hydrogen at 1.89 ppm, a single line of methylene hydrogen at 4.97 ppm, a double line of methylene hydrogen at 5.40 ppm, and a double line of methylene hydrogen at 7.10 ppm. shows the single resonance line of hydrogen at the 5-position of the thiazole ring. Example 7 (1) α-oximino-β-keto-γ-bromobutyric acid ethyl ester 238Tf9 was dissolved in 10 ml of ethanol, and N-(β・β・β-trichloroethoxycarbonyl)thiourea 251rf9 was added thereto for 6 hours. Heat to reflux.

After cooling, the reaction mixture was heated with 50 ml of chloroform, washed with water, and dried over magnesium sulfate. After chloroform is distilled off under reduced pressure, the residue is purified by silica gel column chromatography to obtain 2-(β·β·β-trichloroethoxycarbonylamino)thiazol-4-yl-α-oximinoacetic acid ethyl ester. Yield: 160 pounds. The nuclear magnetic resonance spectrum (100MHz, in deuterium chloroform) of this product is 1
．． The triplet and quartet of ethyl groups are shown at 35 and 4.36 ppm, the single line of trichloroethoxy group is shown at 4.87 ppm, and the single line resonance line of hydrogen at the 5-position of thiazole is shown at 7.94 ppm.

') (2) Dissolve 2.0y of 2-(β・β・β-trichloroethoxycarbonylamino)thiazol-4-yl-α-oximinoacetic acid ethyl ester in 50ml of 10% hydrochloric acid ethanol, and add 50% palladium on carbon. 0.5
Add 7 and absorb hydrogen gas while shaking.

The reaction stops when 90 ml of hydrogen gas is absorbed.
When 1.57 ml of 5% palladium on carbon is added to absorb hydrogen gas, an additional 170 ml is absorbed.

At this point, the reaction is stopped, the reaction mixture is filtered to remove insoluble matter, and the filtered solution is concentrated under reduced pressure.

The residual solid matter is washed with ether and then collected. This solid substance was dissolved in 5 ml of water, neutralized with a 10% aqueous sodium bicarbonate solution, and extracted with chloroform. The chloroform layer was washed with water, dried over magnesium sulfate, and then evaporated under reduced pressure. When the residue was purified by applying it to a silica gel column, 2-(β・β・β-trichloroethoxycarbonylamino)thiazol-4-ylglycine ethyl ester 56
Get 0W9. Nuclear magnetic resonance spectrum of this product (100M
Hz, in deuterium trifluoroacetic acid) are triplet and quartet based on ethyl hydrogen at 1.37 ppm and 4.47 ppm, single line of trichloroethoxy moiety hydrogen at 4.98 ppm, single line of methine hydrogen at 5.82 ppm, 7.74pp
The single resonance line of hydrogen at the 5-position of thiazole is shown in m. (3)
2-(β·β·β-trichloroethoxycarbonylamino)thiazol-4-ylglycine ethyl ester3.
40f7 was dissolved in 50ml of chloroform, 1.2r of triethylamine was added while stirring at room temperature, and then β.
β・β-Trichloroethoxycarbonyl chloride 2.5
Gradually add 0f7.

After 30 minutes, 100 ml of chloroform was added to the reaction mixture, which was then washed with saturated brine, 1N hydrochloric acid, and water, and dried over magnesium sulfate.

After chloroform was distilled off under reduced pressure, n-hexane was added to the residue to solidify it. Ligroin was recrystallized from a mixed solvent of n-hexane.
β·β·β-monotrichloroethoxycarbonyl)glycine ethyl ester 4.117 is obtained. Yield 82%. The nuclear magnetic resonance spectrum (100 MHz, in deuterochloroform) of this product has triplet and quartet based on ethyl groups at 1,21 and 4.60 ppm, and 4.83 and 4.86 ppm.
5. two single wires based on trichloroethoxy groups in m;
A double line of methine hydrogen is shown at 60 ppm, a single line of hydrogen at the 5-position of thiazole is shown at 6.98 ppm, and a double line resonance line of α-amino group hydrogen is shown at 7.38 ppm.

Reference example 1 α-(β・β・β-trichloroethoxycarbonylamino)-α-[2-(β・β・β-trichloroethoxycarbonylamino)thiazol-4yl]acetic acid (237%) was dissolved in 8ml of thionyl chloride. , stir at room temperature for 1.5 hours.

After removing excess thionyl chloride under reduced pressure, 7-acinosephalosporanic acid 183W9 and N-N
Add 5 ml of dimethylacetamide and stir for 4 hours.
After adding 50 ml of ethyl acetate to the reaction mixture, saturated brine was added for decomposition. The ethyl acetate layer was separated, dried over magnesium sulfate, and evaporated under reduced pressure to obtain an oily product of 340Tf1.
Get 9. This oil was dissolved in an excess of 5% aqueous sodium hydrogen carbonate solution, passed through an Amberlite XAD-2 (manufactured by Rohm & Haas) column, and purified by elution with water.
7β-{α-(β·β·β-trichloroethoxycarbonylamino)-α-[2-(β·β·β-trichloroethoxycarbonylamino)thiazol-4-yl]acetamide]cephalosboranate sodium is obtained. The nuclear magnetic resonance spectrum (100 MHz, in heavy trifluoroacetic acid) shows a single line of the acetyl group at the 3-position at 2.25 ppm, and a single line of the acetyl group at the 3-position at 3.7.
2-position methylene hydrogen quartet at 0ppm, 5.00ppm
A single line of the trichloroethyl group is shown at 8.12 ppm, and a single line resonance line of the thiazole ring hydrogen is shown at 8.12 ppm.

Reference example 2 α-(β・β・β-trichloroethoxycarbonylamino)-α-[2-(β・β・β-trichloroethoxycarbonylamino)thiazol-4-yl]acetic acid 1,40
t was dissolved in 25 ml of thionyl chloride and stirred at room temperature for 2 hours.

After removing excess thionyl chloride under reduced pressure, 1.107 g of 7-aminocephalosporanic acid and 25 ml of N--N-dimethylacetamide were added to the residue, followed by stirring at room temperature for 5 hours. After adding 2507n1 of ethyl acetate to the reaction mixture, saturated brine was added and the mixture was shaken.

Separate the ethyl acetate layer, wash with water, and dry with magnesium sulfate. After distilling off the solvent under reduced pressure, 100 ml of 90% formic acid was added to the remaining oil to dissolve it, and 5.0 ml of zinc powder was added while cooling to 0°C.
Add Oy gradually and stir for 3 hours. Insoluble materials are removed by filtration and concentrated under reduced pressure. After adding 30ml of water to the residue,
After passing hydrogen sulfide gas for 5 minutes, the precipitated insoluble matter is removed by filtration. The P solution is concentrated under reduced pressure, and the residue is dissolved by adding 5% sodium hydrogen carbonate, and purified by passing through an Amberlite XAD-2 column. 7β-(
2-aminothiazol-4-ylglycylamide)
Obtain sodium cephalosporanate. Elemental analysis value C
l5Hl6N5O6s2Na・2H20 calculated value: C3
7. ll;H4. l5% Actual value: C37. O9; H3.93%

Claims (1)

[Claims] 1 General formula ▲ Numerical formulas, chemical formulas, tables, etc. Indicates an optional amino group. ] A 2-aminothiazol-4-ylglycine derivative or a salt thereof.