JPH0368035B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to surun 2β-chloromethyl-2α-methylpenam-3α-carboxylic acid, a pharmaceutically acceptable salt or easily hydrolysable ester thereof. These compounds are useful as inhibitors of β-lactamases. Extensive research has been conducted for β-lactamase inhibitors based on the presumed association between the resistance exhibited by β-lactam antibiotics to certain bacteria and the ability of these bacteria to produce β-lactamase. It's here. Clavulanic acid is an example of this type of compound that is currently being extensively studied. Another β-lactamase inhibitor, in its acid form, has the formula European Patent Application No. 2927
(issued July 11, 1979). formula No. 4,036,847, No.
Nos. 4009159, 3993646, 3989685 and 3954732
Disclosed in the issue. U.S. Patent No. 4,155,912 has the formula 2-penem-3-carboxylic acid compounds and their esters and salts have been described (Farmdoc Abstracts 82090A, 10336B and
44337B). formula (CP-45899) is an irreversibly acting β-lactamase inhibitor with excellent solution stability. It has weak antimicrobial activity and enhances the in vitro and in vivo activity of ampicillin against β-lactamase producing bacteria [Antimicrobial Agents and
Chemotherapy, 14 , 414-419 (1978);
Aswapokee et al. J. Antibiotics 31(12), 1238−1244
(December 1978) and Derwents Farmdock
See Abstracts 89627A and 73866B]. B. J. Antibiotics, 33(10), 1183−1192 (1980).
Baltzer et al.'s Mutual Pro−Dragsof β−Lactam
In a paper titled Antibiotics and β-Lactamase Inhibitor, the principle of the combination of β-lactam antibacterial and β-lactamase inhibitory properties in one molecule, which act as precursor drugs for the two active elements, states that ampicillin and mecillinam, respectively, are - The case of conjugation to the lactamase inhibitor penicillanate sulfone is illustrated by its conjugated esters 3 and 4. It has been shown that these esters are very well absorbed from the human gastrointestinal system and then undergo hydrolysis to release the active ingredients. The result is high blood and tissue concentrations of antibiotic and β-lactamase inhibitor in a balanced ratio. This advantage of mutual precursor drugs over simple combinations is discussed. Esters 3 and 4 described herein have the following structures. British Patent No. 2044255 (issued October 15, 1980)
General formula 1 that was not known until now [In the formula, R ₁ is phenyl, 4-hydroxyphenyl, 1,4-cyclohexadienyl or 3-
Represents thienyl; _R2 represents a primary amino group or carboxy group; _R3 is a hydrogen atom, or lower alkyl, aryl, or aralkyl; A includes a β-lactam ring and a carboxy group, and is bonded via a carboxy group Compounds having the following β-lactamase inhibitor group are described and are the subject of this British patent. The above-described compounds are useful in the treatment of bacterial infections and have particularly potent activity against β-lactamase producing bacteria. See also Farmdoc Abstractg 60773C and 60776C. The present invention is based on the formula [R ⁰ is hydrogen, a salt-forming group, or a physiologically hydrolyzed ester-forming group] or a pharmaceutically acceptable salt of the acid or an easily hydrolyzable ester of the acid. . Pharmaceutically acceptable salts include non-toxic metal salts such as sodium, potassium, calcium and magnesium salts; ammonium and substituted ammonium salts such as trialkylamines (e.g. triethylamine), procaine, dibenzylamine, N-benzylamine, β-phenethylamine,
1-Ephenamine, N,N'-dibenzyl-ethylenediamine, dehydroabiethylamine, N,
N'-bis(dehydroabiethyl)ethylenediamine, N-(lower)alkylpiperidines (e.g. N-ethylpiperidine) and other amines that have been used to form pharmaceutically acceptable salts of penicillins and cephalosporins. There are ammonium salts of non-toxic amines. The most preferred salts are the alkali metal salts (ie, sodium and potassium salts) and ammonium salts. Easily hydrolyzable esters or physiologically hydrolyzable esters refer to pharmaceutically acceptable esters known in the art that hydrolyze in vivo to form free acids. Examples of these esters are phenacyl, acetoxymethyl, hivaloyloxymethyl, α-acetoxyethyl, α-
Acetoxybenzyl, α-hyvaloyloxyethyl, phthalidyl (3-phthalidyl), indanyl (5-indanyl), methoxymethyl, benzoyloxymethyl, α-ethylbutyryloxymethyl, propionyloxymethyl, valeryloxymethyl, isobutylene An example is ryloxymethyl. Preferred esters are acetoxymethyl, pivaloyloxymethyl, methoxymethyl, phthalyl, and 5-indanyl. The invention also provides formulas as desired products. [R ⁰ is hydrogen, a salt-forming group, or a physiologically hydrolyzable ester-forming group] A method for producing an acid having the following formula, a pharmaceutically acceptable salt of the acid, or a physiologically hydrolyzable ester of the acid. The method is a Formula (A) (wherein R ¹ is benzyl or substituted benzyl) is catalytically hydrogenated using a noble metal catalyst such as palladium, and then b the hydrogenated product is oxidized to form the desired acid or salt thereof. and when producing an ester of the target compound in which c R ⁰ is a physiologically hydrolyzable ester-forming group, esterifying the acid or a salt thereof to obtain an ester of the acid that is easily hydrolyzable; It consists of a continuous process. The invention also provides formulas as desired products. [R ⁰ is hydrogen, a salt-forming group or a physically hydrolyzable ester-forming group] or a pharmaceutically acceptable salt of the acid or a physiologically hydrolyzable ester of the acid. It provides one method, which is a formula Oxidation of the ester with KM _o O ₄ , H ₂ O ₂ or other similar peroxides or peracids gives the formula b reacting the ester sulfoxide with a metal in an acid, such as zinc in glacial acetic acid, to produce the desired acid or salt thereof; and c R ⁰ is a physiological hydration The production of the ester of the target compound, which is a degradable ester-forming group, consists of a continuous process in which the above-mentioned acid or its salt is esterified to obtain an ester of the acid that can be easily hydrolyzed. A wide range of oxidizing agents known in the art can be used to oxidize sulfides to sulfones, including alkali metal permanganates such as potassium permanganate and organic peracids such as
-Chloroperbenzoic acid is a particularly convenient reagent. Particularly useful protecting groups for R ¹ are benzyl and substituted benzyl, with 4-nitrobenzyl being particularly good. Benzyl and substituted benzyl groups can be conveniently removed by catalytic hydrogenation. in this case,
A catalytic amount of the compound of formula (A) (R ¹ is benzyl or substituted benzyl) dissolved in an inert solvent is added under an atmosphere of hydrogen or under an atmosphere of hydrogen diluted with an inert gas such as nitrogen or argon. Stir or stir in the presence of hydrogenation catalyst. Convenient solvents for this hydrogenation are lower alkanols such as methanol; ethers such as tetrahydrofuran and dioxane; low molecular weight esters such as ethyl acetate and butyl acetate; water; and mixtures of these solvents. However, it is useful to choose conditions that will allow the raw material to dissolve. Hydrogenation is normally carried out at temperatures ranging from 0 DEG to 60 DEG C. and pressures ranging from about 1 to about 100 kg/cm ^@2 .
The catalysts used in this hydrogenation reaction are of the type well known in the art for this type of transformation, and include, for example, noble metals such as nickel, palladium, platinum and rhodium. The catalyst is about 0.01 to about
It is present in an amount of 2.5% by weight, preferably about 0.1-1.0% by weight. In many cases, the catalyst is supported on an inert support, and a particularly good catalyst is palladium on an inert support such as carbon.
The reaction mixture is normally used as a buffer and the reaction is carried out at a pH in the range of about 4 to 9, preferably 6 to 8. Borate and phosphate buffers are commonly used. Typically the reaction is run for about 1 hour. The following examples are provided to illustrate the preparation of representative compounds of the invention and are not intended to limit the invention. Example 1 Potassium 2β-chloromethyl-2α-methylpenam-3α-carboxylate (BL-
P2013) manufacturing 6α-Bromopenicillanic acid S-dulfoxide (1) 1 6α-Bromo-penicillanic acid N,N'-dibenzylethylenediamine salt [G. Signarella et al., J.Org.Chem.27, 2668] in 330 ml of methylene chloride.
(1962) and E. Eppard's Nature201,
1124 (1964)] Dissolve 30 g (37.5 mmol).
Cool to 0°C and stir. 2. When 13 ml (156 mmol) of concentrated hydrochloric acid is slowly added to a methylene chloride solution, precipitation of dibenzylethylenediamine HCl salt (DBED.HCl) occurs within 1 minute. Stir the slurry for 10 minutes at 0-5°C. 3 Remove the DBED-HCl precipitate by passing through pre-coated diatomaceous earth (“Dicalite”) and wash the cake with 150 ml of methylene chloride. The process must be completed as quickly as possible. It is better to avoid soaking in acidic methylene chloride solution for a long time. There is a problem because the precipitate is too fine. It is convenient to add a super-aid to the slurry. 4. Wash the combined methylene chloride solution and wash with 60ml of cold water. Stir for 5 minutes and discard the aqueous phase. The pH of the washing liquid is
It is 2.0−2.3. 5. Concentrate the methylene chloride solution containing 6α-bromopenicillanic acid under reduced pressure to a volume of 65-80 ml. Cool and stir the solution to 5°C. 6 While stirring vigorously, add 13 ml of 40% peracetic acid solution (86.9
(mmol) carefully added over 30 minutes.
The reaction is exothermic. Cool with ice melt to bring temperature to 15-18℃
Keep it. After adding 10 ml of peracetic acid, dulphoxide begins to crystallize. Stir the slurry at 0-5°C for 2 hours. 7. Wash the filtered white cake in the following order: 5℃ water.
10ml, 0-5℃ methylene chloride 10ml, and finally heptane 15ml. 8. Dry the cake in a 45°C air oven to constant weight, approximately 6-10 hours is sufficient. A pink color develops when heated for a long time. The weight of 1 is approx.
16.26g (yield 73.24%). 9 The reaction mixture and final product were 15 toluene/
Can be tested by TLC using 4 acetone/1 acetic acid (HAC) or 8 acetone/8 methanol/3 toluene/1 HAC solvent system. The final product needs to be analyzed by NMR and IR. p-Nitrobenzyl 6α-bromopenicillanate s-dulfoxide (2) 6α-bromopenicillanate s in 100 ml of acetone
-7.5 g (0.041 mol) of potassium 2-ethylcaproate were added to a solution of 12 g (0.04 mol) of dulfoxide. This salt was collected by excess, washed with cold acetone, and air-dried to obtain a total amount of 10 g. The crystalline potassium salt was dissolved in 75 ml of dimethylacetamide and 7.8 g (0.04 mol) of p-nitrobenzyl bromide was added. Solution at 23℃
The mixture was stirred for 24 hours, diluted with 500 ml of water, and extracted with ethyl acetate. The ethyl acetate layer was washed with water four times and dried over anhydrous magnesium sulfate. Solvent at 35 °C (15
mm) evaporated to give an oil, which crystallized. Slurry light brown crystal 2 with ether and collect 9 g (70
g) was obtained. Melting point 124-125°C (decomposition). Analytical values for C ₁₅ H ₁₅ BrN ₂ O ₆ S: Calculated values: C, 41.98; H, 3.05; N, 6.52. Measured values: C, 42.00; H, 3.48; N, 6.98. IR (KBr): 1800 ( s), 1740(s), 1610(w),
1520(s), 1450(m), 1350(s), 1060(m), 74
0
(m) cm ^-1 .H-NMR (60mHzDMSO): δ1.22 (s, 3H), 1.6 (s, 3H), 4.67 ((s, 1H)
,
5.2 (d, J~1-5Hz, 1H), 5.45 (s, 2H),
5.68 (d, J~1-5Hz, 1H), 7.5-8.5 (m,
4H) p-nitrobenzyl 2β-chloromethyl-2α-
Methyl-6-bromopenam-3α-carboxylate (3) p-nitrobenzyl in 120 ml of anhydrous dioxane
6α-bromopenicillanate s-dulfoxide (2)
A solution of 5 g (0.012 mol) of quinoline 1.5 g (0.012 mol) and benzoyl chloride 1.6 g under nitrogen
(0.012 mol) and heated under reflux for 4 hours. solution
It was diluted with 600ml of water and extracted with ethyl acetate. The extract was washed with 5% sodium bicarbonate solution, 5% phosphoric acid solution and finally water. The organic layer was dried over anhydrous magnesium sulfate and evaporated at 35°C (15 mm) to give an oil. Collect the crystals that crystallized from the oil, wash with ether, and then with cold toluene.
%) Got. Melting point 130-135°C (decomposition) Analysis for C ₁₅ H ₁₅ ClBrN ₂ O ₅ S: Calculated: C, 40.06; H, 3.14; N, 6.23. Measured: C, 40.19; H, 3.12; N, 6.75. IR (KBr): 1792 (s), 1740 (s), 1610 (w),
1520 (s), 1353 (s), 1280 (m), 1025 (w),
990 (w), 750 (w) cm ^-1 . NMR (60 mHz, DMSO): δ1.45 (s, 3H), 3.5
-4.3 (m, 2H), 5.05 (s, 1H), 5.42 (s,
2H), 5.5 (d, J ~ 1.5Hz, 1H), 5.62 (d,
J ~ 1.5Hz, 1H), 7.5-8.5 (m, 4H). p-Nitrobenzyl 2β-chloromethyl-2α-
Methylpenam-6α-carboxylate sulfoxide (4) p-nitrobenzyl 3β in 50 ml of methylene chloride
-Chloromethyl-2α-methyl-6α-bromopenam-3α-carboxylate (3) 1 g (0.0022 mol)
A solution of 473 mg of m-chloroperoxybenzoic acid
(0.0022 mol) at 23°C for 3 hours. 33℃
The methylene chloride was evaporated to 20 ml using (15 mm) and the concentrate was diluted with 50 ml of heptane ("Skellysorb B").
After decanting the solvent and slurrying the residue with ether, 250 mg of (4) was crystallized. (Yield 24%) Melting point
Analysis for C ₁₅ H ₁₄ BrClH ₂ O ₆ S: Calculated: C, 38.68; H, 3.02; N, 6.02. Measured: C, 39.14; H, 3.13; N, 5.96. IR (KBr): 1800 (s), 1760 (s), 1520 (s),
1350 (s), 1200 (s), 1050 (m), 830 (w),
740 (w) cm ^-1 .H−NMR (60 mHz・DMSO):
δ1.32 (s.3H), 3.8−4.5 (m, 2H), 4.97 (s.
1H), 5.25 (d, J ~ 1.5Hz, 1H), 5.45 (s,
2H), 5.6 (d, J~1.5Hz, 1H), 7.5−8.5
(m, 4H) Potassium 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone (5) (BL-
P2013) p-Nitrobenzyl 2β- in 150 ml of ethyl acetate
Chloromethyl-2α-methyl-6α-bromo-penam-3α-carboxylate sulfoxide (4) 7g
(0.015 mol) was added a suspension of 4 g of 30% palladium on diatomaceous earth ("Ceride") and 2.8 g of sodium bicarbonate in 150 ml of water. Mixture at 50psi
Hydrogenation was carried out for 3 hours. The catalyst was separated, the aqueous layer was separated, and 1.5 g of potassium permanganate was added to 50 ml of water. The mixture was stirred for 1 hour and 250 mg of sodium bisulfite was added. The mixture was filtered, the pH of the solution was adjusted to 2 with concentrated hydrochloric acid, and the solution was freeze-dried in vacuum to obtain a white amorphous powder. The solids were extracted with ethyl acetate and evaporated to a volume of 20 ml with 100 ml of heptane (“Skellysorb B”).
diluted in ml. A white hygroscopic solid, 2β-chloromethyl-2α-methylpenam-3α-carboxylic acid sulfon, was collected. Dissolve acid in acetone to form solid 2-
Potassium ethyl caproate was added. Filtration of the precipitated crystalline white salt gave 170 mg of 5. Melting point＞
140℃ (decomposition). Analysis for C ₈ H ₇ ClKNO ₅ S・2H ₂ O: Calculated: C, 28.27; H, 3.24; N, 4.12. Measured: C, 28.27; H, 3.69; N, 3.84. IR (KBr): 1790 (s), 1770 (m), 1620 (s),
1460 (m): 1370 (s), 1310 (s), 1200 (s),
1140 (s), 955 (m), 740 (m) cm ^-1 . H-NMR (100 mHz, D ₂ O): δ1.68 (s, 3H),
3.2-3.9 (m, J~2Hz, J~4Hz, J~6
Hz, 2H), 4.0-4.4 (m, 2H), 4.3 (s, 1H),
5.02 (dd, J~4Hz), J~2Hz, 1H). Example 2 Bivaloyloxymethyl 2β-chloromethyl-
2α-Methylpenam-3α-carboxylate sulfone One equivalent of triethylamine was added to 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone in dimethylformamide and stirred to form a solution. A solution of bromomethyl pivalate (1 equivalent) in dimethylformamide was then added, the mixture was stirred at room temperature, and the filtrate was poured into ice water. The separated solid was filtered, washed with water, and dried to give the title ester. If equimolar amounts of chloromethyl acetate, chloromethyl methyl ether, chloroacetone and phenacyl bromide are used in place of the bromo-methyl pivalate used in the above process, respectively, the same acids acetoxymethyl, methoxymethyl, acetonyl and phenacyl can be used. Nasil esters can be produced respectively. Example 3 Pivaloyloxymethyl 2β-chloromethyl-
2α-Methylpenam-3α-carboxylate sulfone BL-P2024 BL-P2013(5) 14.6g in 200ml of acetone
(0.0487 mol) was stirred and added with 4 ml of a 10% aqueous sodium iodide solution, and the mixture was refluxed on a steam bath. Redistilled chloromethyl pivalate (boiling point Hg 7 mm) was added to this refluxing suspension.
℃) 14.8 ml (0.1 mol) was added at once. The mixture was stirred and refluxed for 3 hours and then cooled to room temperature (22°C). The crystalline solid was filtered, washed with 3 x 30 ml of acetone, and the combined liquid was evaporated in vacuo below 22°C to give an oil. Take this oil in 500 ml of ethyl acetate, wash once with 200 ml of water and once with ₄ saturated Na ₂ SO solutions, and remove with carbon 2 for decolorization.
The mixture was cooled in an ice bath while stirring with g. After 20 minutes, the mixture was passed through a layer of diatomaceous earth (Dicalite) and the layer was washed with 4 x 100 ml of ethyl acetate. The combined liquids were concentrated in vacuo at 22°C to obtain an oil. The oil was then further concentrated at 22° C. below 1 mm Hg to remove most of the residual chloromethyl pivalate. Then add 2x50ml of oil
of n-pentane and left in a cool place (approximately 10° C.) under n-pentane over the weekend. The crystalline solid was triturated to a solid powder in a 4:1 mixture of ether-n-pentane and then filtered.
It was washed sequentially with pentane (1:1) and pentane and air-dried. 13.37 g of bivaloyloxymethyl 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone after vacuum drying over P ₂ O ₅ for 4 hours
(yield about 75%). Melting point 93-95°C Analysis for C ₁₄ H ₂₀ ClNO ₇ S: Calculated: C, 44.03; H, 5.27; N, 3.67. Measured: C, 44.11; H, 5.08; N, 3.85. Example 4 Potassium 2β -chloromethyl-2α-methylpenam-3α-carboxylate sulfone (BL-
Recrystallization of P2013) 1 ml of water was added at once to a mixture of 20 ml of n-butanol and 1 g of BL-P2013 (5), and the mixture was shaken in a separating filter to obtain a pale yellow solution. The clear liquid was poured into a long piece of paper, and the flask and paper were washed with 10 ml of a 9:1 n-butanol-H ₂ O solution.The combined liquid was further diluted with 20 ml of n-butanol. The resulting solution was placed in a round bottom flask on a roto-vap and evaporated to about half its original volume under reduced pressure. The pure white crystalline product was filtered, washed with 6 x 10 ml of acetone, and air-dried. The yield was 810 mg, and 800 mg (80%) was obtained by vacuum drying for 6 hours under Hg of less than 1 mm on P ₂ O ₅ . melting point
215℃ (decomposition). Analysis for C ₈ H ₉ ClNO ₅ SK・1H ₂ O: Calculated value: C, 29.67; H, 3.39; N, 4.63; Cl, 10.94; KFH ₂ O, 5.56. Measured value: C, 29.23; H, 3.38; N, 4.49; Cl, 10.74; KFH ₂ O, 5.74. This recrystallization process produces a crystalline monohydrate distinct from the essentially anhydrous starting material. Reference example 1 Chemical Abstracts 27 , 2427 ¹ and 22 , 382 ⁸
and see GB299064. a In a solution of compound 5 (0.1 mol), potassium bicarbonate (0.3 mol) and tetrabutylammonium hydrogen sulfate (0.01 mol) in 200 ml of dichloromethane-water (1:1), dichloromethane was added while keeping the reaction temperature below 30°C. A solution of chloromethyl chlorosulfate (0.115 mol) in 40 ml was added dropwise. After the addition was complete, the mixture was stirred for 30 minutes at room temperature, the organic phase was separated and the aqueous phase was extracted with 50 ml of dichloromethane. The organic phases were combined and diluted with Na ₂ The residue dried with SO and evaporated in vacuo was dissolved in 150 ml of ether. After adding diatomaceous earth, the insoluble matter was separated and the liquid was evaporated in vacuo to give compound 7. b 51.5 g of compound in 12 ml of dimethylformamide 1.6 g of bischloromethyl sulfate was added to the suspension and the mixture was stirred at room temperature for 45 minutes.This was diluted with 50 ml of ethyl acetate and the mixture was washed with water, aqueous sodium bicarbonate, dried and evaporated in vacuo to give compound 7.
was obtained as oil. c Compound 5 in 7.5 ml of dimethylformamide
(0.005 mol) of triethylamine (0.007 mol) and chloroiodomethane (0.030 mol)
mol) was added and the mixture was stirred at room temperature for 4 hours.
After diluting this with 30ml of ethyl acetate, the mixture was
The mixture was washed with 10 ml of water, further washed with 5 ml of saturated aqueous sodium chloride solution, dried, and evaporated in vacuo to obtain compound 7 as an oil. d Compound 5 (0.15 mol) in 750 ml of acetonitrate,
Chloroiodomethane (1.5 moles) was added to a mixture of silver nitrate (0.15 moles) and silver oxide (7.5 g). After stirring at room temperature for 48 hours, the silver salts were separated and the solution was evaporated to dryness in vacuo. Dilute the residue with ethyl acetate
The solution was dissolved in 200 ml, washed with a saturated aqueous sodium chloride solution, dried, and evaporated in vacuo to obtain Compound 7. Compound 7 and other intermediates and final products of the invention were purified, if necessary, on a "Sephadex" LH20 using chloroform-hexane (65:
35) or by column chromatography using, for example, Mallinckrodt CC-7 and hexane-
Ethyl acetate (3:2) or ethyl acetate-petroleum ether (8:2 or 7:3 or 1:9 or 15:85)
or ethyl acetate-n-hexane (4:6 or 3:
1) or ethyl acetate-hexane (1:3 or 1:
1 or 4:1) or by silica gel chromatography using cyclohexane-ethyl acetate (1:1). Thin layer chromatography is also convenient. "Sephadex" is cross-linked dextran 2-(dimethylamino)ethyl 2-[[2-(diethyl-amino)ethyl]diethylammonio]ethyl ether chloride hydrochloride epichlorohydrin.
(See Merck Index, 9th edition, entry 7337) Compound 7 (0.2 mol) and sodium iodide (0.3
mol) was stirred in 150 ml of acetone at room temperature for 18 hours. The resulting suspension was cooled to approximately 0°C, and a saturated aqueous sodium bicarbonate solution was added with stirring to adjust the pH to approximately 0°C.
Adjusted to 7.2. After titrating with 0.5M aqueous sodium sulfate solution to make it colorless, dilute the solution with 150% water while stirring.
ml of acetone-water (1:1) solution,
Washed with x20 ml of isopropanol and 2 x 20 ml of ether and dried. Ampicillin can be converted to compound 9 by the use of methyl acetoacetate in the method of US Pat. No. 3,316,247. Then 1 liter of dimethylformamide
Compound 8 (0.5 mol) was added to the solution of Compound 9 (0.57 mol) at 5° C. while stirring. 5℃
After stirring for 15 minutes, the reaction mixture was poured into an ice-cooled mixture of 4 liters of ethyl acetate and 2 liters of a saturated aqueous calcium chloride solution while stirring. The organic layer was separated, washed with 2 x 500 ml of saturated aqueous calcium chloride solution and evaporated in vacuo to ca. 1 to give a concentrated solution of compound 10. Add water to this liquid
A solution of compound 11 was obtained by adding 500 ml and 500 ml of n-butanol, and adding 4N hydrochloric acid dropwise with stirring until the amino protecting group was removed. After the acid addition was completed, 1 liter of ether and 500 ml of water were added to the mixture while stirring, the aqueous phase was separated, and the organic phase was extracted with 800 ml of water. After washing the combined aqueous extracts with ether 1, add sodium chloride.
640 g and 2 dichloromethane were added and the mixture was stirred for 15 minutes. The organic phase was separated, the aqueous phase was extracted with 1 portion of dichloromethane, and the combined extracts were dried with MgSO ₄ to a
Evaporation under reduced pressure to 600 ml gave a concentrated solution of compound 11.
200 ml of 2-butanone was added thereto and the mixture was cooled to precipitate a solid 6-[(R)-2-amino-2-phenylacetamide]-3.3-dimethyl-7-oxo-4-
Thia-1-azabicyclo[3.2.0]-heptane-2
-Carbonyloxymethyl 2β-chloromethyl-
2-α-Methylpenam-3α-carboxylate sulfone (11) was over-collected. Reference example 2 6-[(R)-2-amino-2-p-hydroxyphenyl-cetamido]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane- 2-Carbonyloxymethyl/2β-chloromethyl-2-α-methylpenam-3α-carboxylate sulfone In the method of Reference Example 1, ampicillin is replaced with amoxicillin to obtain the formula: The title compound represented by was prepared. Example 5 50 g (0.375 mol) of phthalide recrystallized as described in U.S. Pat. No. 3,860,579 and N-bromosuccinimide (0.375 mol) recrystallized as described in U.S. _Pat . Refluxed. The mixture was cooled to about 15° C., the succinimide was removed and washed with about 100 ml CCl ₄ . The combined _four CCl phases were concentrated in vacuo to about 150 ml, and the resulting solid 3-bromophthalide was collected by over-collection to about 50 ml.
It was washed with CCl ₄ and air dried to yield 54 g. This was recrystallized from boiling cyclohexane to give 50 g. Melting point 84-86℃. A part-solution suspension of 2.3 g (0.0075 mol) of compound 5 (BL-P2013) in 20 ml of dimethylformamide (DMF; passed through 3 Å molecular sieves and dried for at least 3 weeks) was mixed with stirring into 3-bromophthalide ( 12) Add 1.7 g (0.008 mol) and heat the mixture at 22°C.
Stir for hours. The resulting mixture was poured into a mixture of 200 ml of ice-cold water and 200 ml of ice-cold ethyl acetate (the flask was washed with a small amount of ethyl acetate) and the mixture was shaken. The organic solvent phase was then separated and diluted with 100 ml of ice-cold water.
I washed it several times. Saturate the ethyl acetate phase with Na ₂ SO ₄
The residual oil, which had been washed once with an aqueous solution, dried over Na ₂ SO ₄ while still cooled, filtered and evaporated to dryness in vacuo, was washed twice with 25 ml of methylcyclohexane as "Skerisorb B" (boiling point 60-68 °C, essentially The mixture was triturated twice with 25 ml of n-hexane and four times with 25 ml of n-hexane, and then air-dried to obtain 132.5 g of an almost white solid compound.
The product was dried over P ₂ O ₅ under 1 mm Hg to yield 132.5 g of compound. Melting point 104℃ (decomposition),
Its purity was estimated to be 85-95%. Analysis _{for C16H14ClNO7S} : Calculated: C, _51.61 ; H, 3.79; N, 3.77; Cl, 9.53 Measured: C, 52.59; H, 4.67; N, 3.21; Cl, 7.73; _KFH2O , 0.27. Example 6 Pivaloyloxymethyl 2β-chloromethyl-
2α-Methylpenam-3α-carboxylate sulfone Potassium 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone hydrate 1 g
(0.0031 mol) and 1 g of 3A molecular sieve was stirred in 15 ml of dimethylacetamide at 23° C. for 2 hours. 470 mg (0.0031 mol) of pivaloyloxymethyl chloride was added to this mixture and stirring was continued for 18 hours. The molecular sieve was separated, the liquid was diluted with 100 ml of water, and extracted with ethyl acetate. Ethyl acetate was washed with water 9 times, dried over anhydrous magnesium sulfate, and dried at 30°C (15
The oil obtained by removing the solvent with
Chromatography using CC-7 (methylene chloride 8, ethyl acetate 2) showed 1 spot Rf.5. The residue was crystallized from heptane ("Scherisorb B") to give 100 mg of pivaloyloxymethyl 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone. Melting point 94-95°. Analysis: Calculated: C, 44.03; H, 5.27; N, 3.67. Measured: C, 44.20; H, 5.24; N, 3.63. NMR and IR spectra were consistent with the structural formula. Example 7 Sodium 2β-chloromethyl-2α-methylpenam-3α-carboxylate sulfone To a solution of 500 ml of BL-P2013 (potassium salt) in 5 ml of water and 10 ml of ethyl acetate was added 2NHCl with stirring until the pH reached 1. (Stir vigorously in an ice bath.)
The mixture was saturated with Na ₂ SO ₄ , the aqueous phase was separated and the organic phase was dried in ice over Na ₂ SO ₄ and then filtered to a wet pH.
50% NaEH in anhydrous n-butanol until paper becomes neutral.
(Sodium 2-ethylcaproate) was added dropwise.
Even with vigorous stirring, no product crystallized. Next, the oil that had been concentrated in vacuo was dissolved in 5 ml of acetone and stirred, but no crystals formed. Ether was added to the cloud point, but no crystals formed, so the oil was concentrated in vacuo on a rotovap, and the resulting oil was dissolved in ethyl acetate and 1 drop of water. was added and stirred, but no crystallization occurred. The residue was then triturated with 5 ml of n-butanol to obtain 200 mg of amorphous white powder.
Wash this with ether, air dry, and vacuum dry on P ₂ O ₅ .
Final sodium 2β-chloromethyl-2α after 24 hours
-Methylpenam-3α-carboxylate sulfone 180mg was obtained. Undefined at decomposition temperature of 100℃ or higher. Analysis for C ₈ H ₉ ClNO ₅ SNa: Calculated: C, 33.10; H, 3.13; N, 4.89. Measured: C, 33.20; H, 3.69; N, 4.44. KFH ₂ O, 4.04 Example 8 Potassium 2β -Chloromethyl-2-methylpenam-3-carboxylate sulfone (BL-
P2013) Water 10, Sodium hydrogen carbonate 130g (1.25mol)
and p-nitrobenzyl 6α-bromo-2β-chloromethyl-2-methylpenam-2 dissolved in ethyl acetate 5 in a mixture of 200 g of 10% Pd on BaSO ₄
A solution of 272 g (0.565 mol) of 3-carboxylate sulfone was added. The mixture was hydrogenated at 40° C. and 1 Kg pressure. After 5 hours, hydrogen absorption became very slow, so hydrogen was added until 200 g of 10% Pd on BaSO ₄ was added until no rapid hydrogen absorption was observed. The slurry was filtered through a layer of diatomaceous earth ("Celide"), the layer was washed with water, and the aqueous phase was washed with 3 portions of ethyl acetate. Add 3 ethyl acetate to the aqueous solution and mix 10
At ℃, 150 ml of 12NHCl was added to adjust the pH to 1.5. The organic phase was separated and the aqueous solution was saturated with Na ₂ SO ₄ .10H ₂ O and 2x
1 and extracted with ethyl acetate. The extracts were combined and dried over magnesium sulfate. 2N 2-ethyl-caproate potassium solution in butanol after removing the desiccant.
260ml was added at 0°C. After stirring for 2 hours at 0 degrees, potassium 2β-chloro-methyl-2-methylpenam-3-carboxylate sulfone (BL-P2013) was collected and vacuum dried at room temperature to obtain a yield of 134.8 g (about 70%). Reference example 3 p-nitrobenzyl 6α-bromopenicillinate sulfoxide Method: - Add 200ml of N,N-dimethylacetamide
6α-Bromopenicillanic acid sulfoxide 44g
(0.148 mol) and further triethylamine 20.5 ml
(0.148 mol) and p-nitrobenzyl bromide 38.2 g
(0.177 mol) was added and stirred at 22° for 20 hours. The reaction mixture was poured into 1 portion of water and extracted with 3×300 ml of methylene chloride. The combined methylene chloride extracts were washed with 200 ml of 5% aqueous sodium bicarbonate solution, passed over sodium sulfate, and dried at 5°C for 30 minutes.
The liquid was filtered and evaporated in vacuo, the resulting residue was diluted with ether, the solid was filtered and dried to give 54 g of p-nitrobenzyl 6α-bromopenicillinate sulfoxide.
I got it. Yield 85%. NMR was consistent with the structural formula.
The yield of this step was the same as in the K-salt esterification step. K-salt production (yield of this process 85-
90%) is not necessary. Reference Example 4 Production of p-nitrobenzyl 6α-bromobenicylanate sulfoxide 6α- to N,N-dimethylacetamide 4375
Bromopenicillanic acid (s) dulfoxide 873.0g
(2.95 mol) and stirred while keeping the internal temperature below 35°C. Triethylamine 293g (2.95 mol)
and further added 764 g of p-nitrobenzyl bromide.
(3.54 mol) was added. The mixture was stirred at room temperature for 5 hours and then left overnight. The reaction mixture was poured into 20ml of water and extracted with 3x7 methylene chloride. Combined organic extracts 5x7
was washed with water and 5% aqueous sodium bicarbonate solution and dried over anhydrous magnesium sulfate. The magnesium sulfate was separated, the liquid was evaporated, the residue was crystallized, diethyl ether from step 4 was added, the crystals were collected, and the crystals were dried at room temperature to give p-nitrobenzyl 6α-.
Bromopenicillanate sulfoxide 1171g (92
%) was obtained. Br18.4% (calculated value 18.53), α _D (0.25% MeOH) + 162°. Reference Example 5 Production of p-nitrobenzyl 6α-bromo-2β-chloromethyl-2-methylpenam-3-carboxylate sulfone Add p-nitrobenzyl 6α-bromo to acetic acid 16
364.6 g (0.812 mol) of 2β-chloromethyl-2-methylpenam-3-carboxylate was added.
Add 282g of KmnO ₄ to 26ml of water while stirring the resulting solution.
(1.78 mol) was added dropwise over 3 hours at room temperature. The mixture was then stirred at room temperature for 1 hour and 37% H ₂ O ₂ was added dropwise until the liquid became colorless. After adding 30 ml of water, the mixture was stirred for 1 hour at room temperature, and the precipitated crystals were collected, washed with 3 x 5 portions of water and 2 x 2 portions of ethanol, and dried under vacuum at room temperature to yield 297 g (76%). α _D (0.5
% _CH2Cl )+75.9゜Example 9 Production of BL-P2013 free acid Add BL− to a mixture of 25 ml of ethyl acetate and 10 ml of water.
800 mg (0.00261 momole) of P2013 potassium salt was added. After all the solids had dissolved, 50% aqueous phosphoric acid solution was added dropwise to the mixture while stirring vigorously until no more precipitate formed from the aqueous layer. The ethyl acetate layer was separated, washed with saturated sodium chloride solution, and dried over anhydrous magnesium sulfate. The desiccant was removed and washed with 10 ml of ethyl acetate. (Washing solution was combined with the original solution) Add "Skerisorb B" to ethyl acetate and the cloud point (approximately 10
ml). Activated carbon (“Darco KB”) in the mixture
500 mg was added and treated, and 15 ml of "Skellysorb B" was added to the separated solution to add the crystal seeds after diluted BL-P2013 was released. After 3 hours at room temperature, the crystalline precipitate of free acid was collected and dried in vacuo over P ₂ O ₅ to give 323 mg (46
%) was obtained. Slowly decomposes at melting points of 100℃ or higher. Analysis for C ₈ H ₁₀ ClNO ₅ S: Calculated: C, 35.89; H, 3.77; N, 5.23; Cl, 13.25 Found: C, 35.88; H, 3.91; N, 5.41; Cl, 13.52 The product is It was found to be unstable when stored at 23°C for several days. Example 10 6α-bromopenicillanic acid sulfoxide 300 g of 6α-bromopenicillanic acid N,N'-dibenzyl-ethylenediamine salt in 3 methylene chloride
(0.75 mol) was added, the suspension was cooled to 5° C., and 130 ml of concentrated HCl was added dropwise over 15 minutes with good stirring. The cake was then filtered through a layer of diatomaceous earth ("ceride") and washed with 3 x 250 ml of methylene chloride. The combined methylene chloride solutions were washed with 2 x 500 ml of water and dried over sodium sulfate for 15 minutes. Sodium sulfate The liquid was separated and the liquid was evaporated under reduced pressure to about 750 ml. This liquid was cooled to 5°C, and 130 ml of 40% peracetic acid was added dropwise with vigorous stirring while maintaining the temperature between 5 and 12°C. The addition was an entirely exothermic reaction. Hot. 5℃ after addition.
The slurry was stirred for 2 hours, and the product was filtered and washed with 100 ml of cold water (5°C) and 100 ml of cold methylene chloride (5°C). Thus, 126 g (57%) of 6α-bromopenicillanic acid sulfoxide was obtained. Melting point: 129°C. IR and NMR spectra were consistent with the desired product. _{Analysis for C8H10BrNO4S} : Calculated: C, 32.44; H, 3.40; N, 4.73. Measured: C, 32.30; H, 3.35; N, 4.71; _H2O , 2.18. Potassium 6α-bromo Penicillinate sulfoxide 126 g (0.43 mol) of 6α-bromopenicillanic acid sulfoxide in acetone 3 and 50% by weight potassium 2-ethylcaproate solution in n-butanol
After adding 162 ml and stirring at 22° C. for 1 hour, the product was filtered, washed with 2×250 ml of acetone, and dried. Thus, 127 g (90°C) of potassium 6α-bromopenicillanate sulfoxide was obtained. Melting point: 185°. IR and
The NMR spectrum was consistent with the desired structural formula. _Analysis for _C8H9BrKNO4S : _Calculated : C, 28.75; H, 2.71; N, 4.19. Measured: C, 29.03; H, 2.78; N, 4.04. p-nitrobenzyl 6α-bromopenicillanate Dulfoxide Potassium in N,N-dimethylacetamide 1
6α-bromopenicillanate sulfoxide 145g
(0.43 mol) was added and set at 22° with stirring.
Add 115 g (0.53 mol) of nitrobenzyl bromide.
Stirred at 22°C for 20 hours. The reaction mixture was poured onto ice 3 and extracted with 3 x 1500 ml of ethyl acetate. Combined extracts 2 x 500
mL of 5% aqueous sodium bicarbonate and dried over sodium sulfate for 30 minutes. The sodium sulfate was removed and the solution was evaporated under reduced pressure, and the residue was treated with dithyl ether 1 to crystallize the product. The crystals were filtered, washed with 2×100 ml of diethyl ether, and dried to yield 162 g (87%) of p-nitrobenzyl 6α-bromopenicillanate sulfoxide. Melting point 111
℃. IR and NMR spectra were consistent with the desired structural formula. Analysis _{for C15H16BrN2O6S} : Calculated: C, 41.78; H, 3.51; N, 6.50; Measured _: C, 41.66; H, 3.45; _N , 6.85; _H2O , 0.69.p- Nitrobenzyl 6α-bromo-2β-chloromethyl-2-methylpenam-3-carboxylate p-dioxane 1 and p-nitrobenzyl 6α
- 70 g (0.16 mol) of bromopenicillanate sulfoside was added, followed by 21.2 ml (0.10 mol) of benzoyl chloride and 21.8 ml (0.19 mol) of quinoline. The reaction mixture was refluxed for 4 hours and then cooled to 22°.
Pour into 2500 ml of H ₂ O and extract with 3 x 800 ml of ethyl acetate. The combined extracts were washed sequentially with 300 ml of 5% aqueous sodium bicarbonate, 300 ml of 5% phosphoric acid, and 300 ml of water. The ethyl acetate solution was passed over sodium sulfate and dried for 30 minutes to remove the sodium sulfate. The residue obtained by evaporating the liquid under reduced pressure was dissolved again in ethyl acetate 1 and evaporated again under reduced pressure to obtain a residue. Add diethyl ether 1 and collect the product to obtain p-nitrobenzyl 6α-bromo-2β-
41 g (57%) of chloromethyl-2-methylpenam-3-carboxylate was obtained. Melting point 132°, IR
The NMR spectrum was consistent with the desired structural formula. Analysis for C ₁₆ H ₁₄ BrClN ₂ O ₅ S: Calculated: C, 40.06; H, 3.14; N, 6.23. Measured: C, 40.62; H, 3.11; N, 6.13. p-nitrobenzyl 6α-bromo- 2β-chloromethyl-2-methylpenam-3-carboxylate sulfoxide p-Nitrobenzyl 6α in 1200ml of methylene chloride
-Bromo-2β-chloromethyl-2-methylpenam-3-carboxylate (51 g (0.11 mol)) was added, followed by 23 g (0.12 mol) of m-chloroperoxybenzoic acid.
mol) was added. The solution was stirred at 22° for 2 hours and evaporated under reduced pressure to give a wet residue. To this, diethyl ether 4
was added, stirred for 1 hour, and left at 10° for 20 hours. The crystallized product was filtered, washed with 2 x 200 ml of diethyl ether, and dried to give p-nitrobenzyl 6α-
Bromo-2β-chloromethyl-2-methylpenam-3-carboxylate sulfoxide 39 g (75
%) Got. Melting point 132°. IR and NMR spectra were consistent with the desired structural formula. Analysis for C ₁₅ H ₁₄ BrClN ₂ O ₆ S: Calculated: C, 38.69; H, 3.03; N, 6.07; Measured: C, 38.98; H, 3.04; N, 5.84; H ₂ O, 0.35. Potassium 2β -Chloromethyl-2-methylpenam-3-carboxylate sulfone (BL-
P2013) To 600 ml of water were added 8 g of 30% Pd on "Celide" and 16 g (0.19 mol) of sodium bicarbonate. p-nitrobenzyl 6α-bromo-2β in 400 ml of ethyl acetate
-Chloromethyl-2-methylpenam-3-carboxylate 32 g (0.69 mol) of sulfoxide was dissolved and added to the slurry. The mixture was hydrogenated on a Parr apparatus at 50p, sci, 22°C for 4 hours. The slurry was filtered through a sintered glass funnel and a layer of "Celide" on top and the layers were washed with 2 x 50 ml of water to separate the combined and aqueous wash layers. Wash the aqueous layer with 200 ml of diethyl ether, cool to 5°, and adjust the pH by adding 40% H ₃ PO ₄ .
12 g of KMnO ₄ in 200 ml of water keeping the temperature between 7.5 and 8.0.
(0.076 mol) was added dropwise over 30 minutes. The addition of KMnO ₄ was stopped when the pink color did not disappear for 5 minutes. The reaction mixture was mixed with a small amount (approximately 50 mg) of sodium bisulfite and after stirring for 30 min the slurry was layered with “celite”. Wash the layer 2x with 50 ml of water. Combine the washing solution with 500 ml of ethyl acetate and stir the layer to adjust the pH.
Adjusted to 1.5 with 2NHCl. The separated aqueous layer was saturated with sodium sulfate. This was extracted again with 2 x 400 ml of ethyl acetate and the combined extracts were dried over sodium sulfate at 5° for 30 minutes. The sodium sulfate was removed and the liquid was evaporated under reduced pressure to obtain a residue. Add this to acetone
To 160 ml were added 160 ml of dissolved diethyl ether and 50% by weight potassium 2-ethyl-caproate in n-butanol so that the solution was neutral to the wet PH paper. The potassium salt of BL-P2013 that crystallized was collected by over-collection, washed with diethyl ether and dried to recover potassium.
2β-chloromethyl-2-methylpenam-3-carboxylate sulfone (BL-P2013) 16g
(76%). The melting point was 202°, and the IR and NMR spectra were consistent with the desired structural formula. Analysis _{for C8H9ClKNO5S} : Calculated: C, 31.42; H, 2.97; N, 4.58; Found _: C, 31.18; H, 2.98; N, 4.51; _H2O , 0.93. Example 11 Piparoyloxymethyl 2β-chloromethyl-
2-Methylpenam-3-carboxylate sulfone (BL-P2024) A suspension of 14.6 g (0.0487 mol) of potassium 2β-chloromethyl-2-methylpenam-3-carboxylate sulfone (BL-P2013) in 200 ml of acetone was mixed with a 10% aqueous sodium iodide solution 4 while stirring.
ml and the mixture was refluxed on a steam bath. To this refluxing suspension, 14.8 ml (0.1 mol) of recrystallized chloromethyl pivalate (boiling point 34° C. at 7 mm Hg) was added at once. The mixture was stirred and refluxed for 3 hours and then cooled to room temperature (22°C). Over-collection of crystallized solids 3
The combined solution was washed with 30 ml of acetone and evaporated under reduced pressure below 22°C to obtain an oil. The oil was taken up in 500 ml of ethyl acetate and washed once with 200 ml of water and once with 200 ml of saturated Na ₂ SO ₄ solution. 2g of decolorizing charcoal while cooling this liquid (ice bath)
It was briefly dried over Na ₂ SO ₄ with stirring. After 20 minutes the mixture was filtered through a layer of "Celide" and the layer was washed with 4.times.100 ml of ethyl acetate. The combined liquid was concentrated under reduced pressure at 22°C to obtain an oil. more oil
Most of the residual chloromethyl pivalate was removed by concentrating at 22° C. under 1 mm of Hg. Then mash the remaining oil with 2 x 50ml of n-pentane for about 10 minutes over the weekend.
C. under n-pentane. The solid crystalline material obtained was then ground to a powder under 40 ml of a 4:1 mixture of diethyl ether-n-pentane. The product was filtered, washed successively with diethyl ether-n-pentane (1:1) and n-pentane, and air-dried.
Dry pivaloyloxymethyl under high vacuum _{over P2O5}
2β-chloromethyl-2-methylpenam-3-carboxylate sulfone (BL-P2024) 13.37g
(approximately 75%). Melting point 93-95℃. Purification of BL-P2024 Approximately 3 g of the crude BL-P2024 obtained above was dissolved in 5 ml of ethyl acetate and placed on a 4.5 x 40 cm silica gel (Marinrot CC-7) tube at 4:1 V/VCH ₂ Cl.
- eluted with ethyl acetate. Separated part with a single spot at Bf0.84 (4 on silica gel plate:
TLC using _1CH2Cl -ethyl acetate, _I2 detection) were combined and concentrated under reduced pressure to obtain 1.38g of crystalline solid. A 900 mg portion of this material was dissolved in 5 ml of ethyl acetate and the resulting solution was filtered and diluted with petroleum ether ("Skellysorb B") to almost the cloud point and then stored at room temperature for 3 days. The generated crystals were collected by excessive collection, washed with petroleum ether, and dried to obtain 560 mg of pure BL-P2024. Melting point 100-101°. Analysis for C ₁₄ H ₂₀ ClNO ₇ S: Calculated: C, 44.03; H, 5.27; N, 3.67. Measured: C, 44.11; H, 5.08; N, 3.85. All temperatures in this application are in degrees Celsius. It is expressed as Example 12 Preparation of BL-P2013 ammonium salt 1 A solution of 250 mg of the free acid of BL-P2013 dissolved in 20 ml of acetone-methanol (volume 1:1) was filtered to obtain a clear liquid. 2. Add 1 ml of ammonium hydroxide (30%, reagent grade) to 10 ml of acetone-methanol (volume 1:1) to make an anhydrous ammonium solution. Add 1 g of anhydrous magnesium sulfate while stirring. Wipe the mixture with paper. "liquid". 3 Add about 2 ounces of “anhydrous ammonium solution” to the solution from method 1.
ml was added slowly and stirred well. 4. 100 ml of diethyl ether was mixed with the mixture of method 3 to precipitate the ammonium salt of BBL-P2013. 5 Separate the white ammonium salt from the solvent and
ml of diethyl ether. 6 The separated powder was dried in a vacuum oven at 35°C overnight. 7 Analysis results as follows: Calculated value: C, 33.7; H, 4.6; N, 9.8. Measured value: C, 33.66; H, 4.63; N, 10.12; Dry by KF. Microscopic examination: Crystalline material. Example 13 Preparation of non-hygroscopic sodium salt of BL-P2013 1 50 mg of the free acid of BL-P2013 was dissolved in 4 ml of acetone-methanol (1:1 volume). It was filtered to obtain a clear liquid. 2 Acetone-methanol (volume 1:1) mixture
A sodium 2-ethylcaproate solution was prepared by dissolving 40mg of sodium 2-ethylcaproate in 10ml. 3. Add 10 ml of the method 2 solution to the method 1 solution and mix well. 4 The mixture of method 3 was mixed with 10 ml of diethyl ether to precipitate the sodium salt of BL-P2013. 5 The white salt was soaked in diethyl ether for 1-2 hours, then separated from the solvent and washed with 3 x 5 ml of diethyl ether. 6 The separated powder was dried in a 30°C vacuum oven overnight. Example 14 Recrystallization of BL-P2013 BL−P2013 400mg with minimum amount of acetone−H ₂ O
(volume 1:1), diluted with 10 ml of acetone, added acetone to make 25 ml, stirred vigorously, and after 30 minutes, collected the crystalline hydrate, washed thoroughly with acetone, air-dried, and vacuum-dried overnight at less than 1 mm of Hg, yielding 280 mg. I got it. Analytical values for C ₈ H ₉ ClNOSK・H ₂ O: Calculated values: C, 29.67; H, 3.39; N, 4.63; Cl, 10.94; H ₂ O, 5.55. Measured values: C, 29.32; H, 3.32; N , 4.44; Cl, 11.31; H ₂ O, 5.90. Example 15 BL-P2013 N,N'-dibenzylethylene-
diamine salt 306 mg (0.001 mol) of BL-P2013 was dissolved in 7 ml of _H2O and added to a solution of 180 mg (0.0005 mol) of N,N'-dibenzyl-ethylenediamine diacetate in 7 ml of water. When the mixture is stirred, the salt crystallizes out, and after about 10-15 minutes, the salt is over-collected and air-dried.
300 mg of N'-dibenzylethylenediamine salt was obtained. This was dissolved in about 10 ml of boiling acetone, diluted with ether to the cloud point, recrystallized, air-dried, and vacuum-dried to obtain 260 mg. Analytical Calculated: C, 51.69; H, 5.42; N, 7.53; Cl, 9.55. Measured: C, 49.39; H, 5.49; N, 7.05; Cl, 8.96; _H2O , 1.23 (KF). Example 16 Chloromethyl ester of BL-P2013 BL−P2013 in a mixture of 50 ml of water and 50 ml of CH ₂ Cl ₂
(5) 15.25g (0.05mol), KHCO ₃ 15g (0.15mol)
and tetrabutyl-ammonium hydrogen sulfate (Aldrich Chemical Company) 1.7 g (0.005 mol)
The mixture was dissolved in 40 ml of CH ₂ Cl ₂ with vigorous stirring.
A solution of 9.5 g (0.0575 mol) of _ClCH2 -O- _SO2Cl was added dropwise. The temperature rose to 26°C and stirring was continued for an additional 30 minutes after the addition (which took approximately 15 minutes). The product crystallized out and about 400 ml more of CH ₂ Cl ₂ was added to bring it into solution.
Separate the _two CH ₂ Cl layers and wash them with 50 ml of CH ₂ Cl ₂ . Combine them, dry over MgSO ₄ and decolorize with carbon (“Darco KB”).
Added 2g. After about 30 minutes, the mixture was filtered and concentrated to about 50 ml, and 150 ml of isopropyl alcohol was added. The remainder of CH ₂ Cl ₂ was removed under reduced pressure. The resulting crystalline precipitate was filtered, thoroughly washed with isopropyl alcohol, and air-dried. Vacuum drying was performed under less than 1 mm of Hg to obtain 8.5 g of chloromethyl 2β-chloromethyl-2-methylpenam-3-carboxylate sulfone (7).
Melting point: 116° (decomposes, darkens above 100°C) Analysis for C ₉ H ₁₁ Cl ₂ NO ₅ S: Calculated value: C, 34.18; H, 3.51; N, 4.43; Cl, 22.43. Measured value: C, 34.16; H, 3.45; N, 4.47; Cl, 22.46; _H2O , 0.33 (KF) Estimated purity range 90-95%. Iodomethyl ester of BL-P2013 To a stirred mixture of 5 g (0.0159 mol) of chloromethyl ester of BL-P2013(7) in 25 ml of acetone, 3 g (0.02 mol) of sodium iodide was added. The resulting slurry was stirred for 17 hours and cooled to about 0°C. Two drops of a saturated aqueous KHCO ₃ solution were added and the mixture was slowly added dropwise to 50 ml of water over a period of 10 minutes. The slurry suddenly changes color from yellow to gray to purple to black, at which point the crystals are immediately collected and cooled with acetone-water (1:
2) Wash with 3 x 10 ml of isopropyl alcohol, diethyl ether and n-pentane sequentially and air dry to obtain 5.55 g of iodomethyl ester of BL-P2013 (8).
(91%). Melting point 118-119°C (decomposed). Estimated purity approximately 90%. Reference example 6 6-[(R)-2-amino-2-phenylacetamide]-3,3-dimethyl-7-oxo-4-
Thia-1-aza-bicyclo[3.2.0]heptane-2-carbonyl-oxymethyl 2β-chloromethyl-2-α-methyl-penam-3α-carboxylate sulfone Deine salt of the above ampicillin (9) (solvated with one molecule of isopropyl alcohol) in 60 ml of acetone.
5.46 g (0.01 mol) of the mixture was cooled in an ice bath, and while stirring, 4.08 g (0.01 ml) of iodomethyl ester (9) of BL-P2013 (8) was added, and the resulting nearly transparent liquid was stirred. The ice bath was removed after 5 minutes and continued for 5 hours. The concentrate, from which most of the acetone had been vacuum removed on a roto-vap, was then dissolved in 200 ml of cold ethyl acetate, 2 x 50 ml of ice-cold water and 2 x 100 ml of saturated
Washed with _Na2SO4 aqueous _solution . Then add ethyl acetate solution
Dry over Na ₂ SO ₄ , filter and remove most of the ethyl acetate in vacuo on a roto-vap. Residue 2x
It was triturated with 200 ml of dry diethyl ether, and the resulting solid was filtered to obtain 5.5 g of pink powder 10. This powder was mixed with a mixture of 50 ml of water, 50 ml of n-butanol and 20 ml of ethyl acetate while stirring.
Add 6NHCl dropwise and keep the pH at 2.5 for about 45 minutes.
Add 2 drops of HCl to maintain pH 2.2-2.5. When the pH no longer rose rapidly, 100 ml of diethyl ether was added and mixed well. The aqueous phase was separated, the organic layer was extracted with 25 ml of water, and the liquids were combined. The aqueous solution was extracted with 50 ml of diethyl ether and the ether was discarded. The aqueous layer was then poured under 100 ml of 2-butanone (methyl ethyl ketone) with vigorous stirring.
Saturated _{with Na2SO4} . The 2-butanone layer was separated and dried over Na ₂ SO ₄ in an ice bath for 30 minutes, filtered and concentrated to near dryness. The residual oil was triturated with n-butanol to form a solid and washed well with ether.
-Wash with pentane, air dry, and vacuum dry over P ₂ O ₅ at less than 1 mm of Hg to obtain crude 6-[(R)-2-amino-2-phenylaceto-amide]-3,3-dimethyl-7.
1.6 g of -oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carbonylmethyl 2β-chloromethyl-2-α-methylpenam-3α-carboxylate sulfone was obtained. IR and
The NMR spectrum matched that of structural formula (11), but the purity was not high. This solid product is at least 40%
Probably about 80% of 6-[(R)-2-amino-2-phenylacetamide]-3,3-dimethyl-7-
Oxo-4-thia-1-azabicyclo [3.2.0]
-heptane-2-carbonyloxymethyl 2β-
It was estimated that it contained chloromethyl-2-α-methylpenam-3α-carboxylate sulfone. Example 17 Improved synthesis method of BL-P2013 This method omits the previous catalytic reduction method and synthesizes BL-P2013.
This is a simplified version of the manufacturing method. (See page 633 of Cephalosporins and Penicillins by Edwin H. Flynn, Academic Press, New York.) 30 g (0.1 mol) of 6α-bromopenicillanic acid sulfoxide (1) are dissolved in 1 ml of dry CH ₂ Cl ₂ followed by 16.2 ml of pyridine ( 0.2 mol) and trichloroethanol
29.8 g (0.2 mol) was added. Next, 20 g (0.1 mol) of dichlorohexylcarbodiimide was added and the mixture was stirred at 22° C. for 16 hours. Dichlorohexylurea began to precipitate and was separated at the end. Add 200 ml of 5% sodium bicarbonate solution, 200 ml of 5% sodium bicarbonate solution,
Washed with 10% phosphoric acid solution and 100 ml of saturated aqueous Na ₂ SO ₄ solution. Pass the organic phase through sodium sulfate at 5°C.
It was dried for 30 minutes, filtered, and evaporated to an oil. Add diethyl ether and stir vigorously to obtain 227 g (63
%) was crystallized. 226.5g (0.062mol) of the compound was added to p-dioxane.
8.5 ml (0.078 mol) of benzoyl chloride dissolved in 500 ml
and 8.75 ml (0.078 mol) of quinoline were added. After the liquid was refluxed for 4 hours, it was poured into 1100 ml of water and the product 3 was extracted into 2 x 400 ml of ethyl acetate. Combine the extracts and add 200 ml of 5% sodium bicarbonate aqueous solution,
Wash with 200 ml of 5% phosphoric acid solution and 200 ml of saturated aqueous sodium sulfate solution and heat over sodium sulfate at 5°C.
The oil (3) obtained by drying and evaporating for 30 minutes was used as it was in the next reaction. Dissolve compound 3 obtained in the previous step in glacial acetic acid 122
Saturate at °C while stirring until the pink color disappears.
KMnO ₄ aqueous solution was added dropwise. (i.e. 1 on the piece of paper
(The droplets turn pink) Then cool until the liquid becomes clear.
30% _H2O2 was added _dropwise . A small amount of white precipitate was formed.
The liquid was poured into 2.5 ml of water and the product 4 was extracted with 3 x 500 ml of ethyl acetate. The ethyl acetate was washed with a 5% aqueous sodium bicarbonate solution until neutral (ie, no rapid foaming occurred upon addition). Drying over sodium sulfate and evaporation gave residue 4. Grind this with “Skerisorb B” at 10℃ every other day to solidify it.
I gained 49.1g. The yield was 28% of the theoretical value for steps 2 and 3 combined. (See US Pat. No. 4,164,497) Slurry 3.75 g of zinc powder in 5 ml of glacial acetic acid.
Cooled to ℃. Add dimethylformamide to this mixture.
Add 43g (0.0057mol) solution to 15ml and 5℃
The mixture was stirred for 2.5 hours. The zinc was removed and the pale yellow liquid was poured into 80 ml of 5% aqueous hydrochloric acid solution. The mixture was extracted with 3 x 25 ml of ethyl acetate, the combined extracts were extracted with 3 x 20 ml of 5% aqueous sodium bicarbonate solution, and the separated ethyl acetate phase was saved. Combine the bicarbonate extracts and place under the ethyl acetate layer.
The pH was adjusted to 1.5 with 2NHCl and saturated with sodium sulfate. Separate the ethyl acetate and add 2x30 ethyl acetate to the aqueous phase.
Extracted in ml. All of the above ethyl acetate was combined, dried over sodium sulfate, and the resulting oil (this is the free acid form of BL-P2013) was dissolved in about 20 ml of acetone and 20 ml of diethyl ether was added. This was supplemented with 50% potassium 2-ethylcaproate (KEH) in dry n-butanol.
was added to make it neutral. Product 5 (BL-P2013) crystallized. After stirring at 22℃ for 30 minutes, filter the
650 mg (37%). Dissolve 50 mg of the sample from step 5 in 0.5 ml of water, add 20 mg of N,
N'-dibenzylethylenediamine (DBED)2
Acetate was added. The crystallized DBED salt of 5 was collected and dried under vacuum over P ₂ O ₅ to give N,N'-dibenzylethylene-diamine-2β-chloromethyl-2-
Methylpenam-3-carboxylate (free acid 5
DBED salt) was obtained. 450 mg of the other sample No. 5 was dissolved in 3 ml of water, and to this was added a solution of 270 mg of DBED2 acetate in 2 ml of water. Upon vigorous stirring, 450 mg of the DBED salt of 5 crystallized. Recrystallization from about 5 ml of boiling acetone yielded 270 mg. Reference example 7 6-[(R)-2-amino-2-p-hydroxyphenyl-acetamide]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane- 2-carbo-oxymethyl
2β-chloromethyl-2-α-methylpenam-
3α-carboxylate sulfone Using the corresponding deine salt of amoxicillin in place of ampicillin used in the method of Reference Example 6, the formula: The above title compound can be prepared. Biological Data Product of Example 1, Formula: Compound 5 having the following will be referred to as BL-P2013. BL-P2013 itself is a very weak antibacterial agent at best, but it inhibits β-lactamases and
It also protects cefuranide and amoxicillin when used in combination with cefuranide and amoxicillin from degradation by β-lactamase producing bacteria in vitro and in vivo.

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[Table] As described above, the compounds of the present invention are useful for improving the effectiveness of β-lactam antibiotics against β-lactamase-producing bacteria by oral and parenteral administration. This amount is from 1/5 to 5 times the amount of the β-lactam antibiotic on a weight basis, preferably the same amount.
By way of example, the compounds of the invention, when used in a 1:1 ratio as shown above, yield B. Fragilis, B.
It significantly improves the activity of cefuranide and amoxicillin against β-lactamase producing strains of anaerobic Bacteroidetes such as Thetaiotaomicron and other species of the genus, and also against resistant Staphylococcus aureus. The compounds of the present invention may be mixed with or simultaneously with a β-lactam antibiotic in the indicated proportions relative to the known common dosage of the antibiotic.
It can be administered either way. Therefore, the ability of the compounds of the present invention to enhance the efficacy of β-lactam antibiotics against certain β-lactamase-producing bacteria makes co-administration with certain β-lactam antibiotics in the treatment of bacterial infectious diseases in mammals, particularly humans. make this compound valuable. For the treatment of bacterial infections, the compounds of this invention can be mixed with beta-lactam antibiotics so that the two drugs can be administered simultaneously. The compounds of the invention can also be administered alone during treatment with β-latatum antibiotics. When the compounds of the present invention or their salts are used to improve the antibacterial activity of β-lactam antibiotics, they are used alone or preferably in combination with standard pharmaceutical carriers and diluents. The compound of the present invention in the acid form or its pharmaceutically acceptable salt form can be administered orally or parenterally. Ester-type compounds of the present invention that are easily hydrolyzed in vivo are most suitable for oral administration. Parenteral administration methods include intramuscular, subcutaneous, intraperitoneal and intravenous administration. When a compound of the invention is used with a carrier or diluent, the carrier or diluent is selected based on the mode of administration. For example, if oral administration is contemplated, the compounds may be used in the form of tablets, capsules, buccal tablets, troches, powders, syrups, elixirs, aqueous solutions and suspensions, etc., according to standard pharmaceutical practices. The proportion of active ingredient to carrier will, of course, depend on the chemical nature, solubility, stability, and potency of the active ingredient and the amount used. However, pharmaceutical compositions may contain from about 5 to about 80% carrier. In tablets for oral administration, carriers commonly used include lactose, sodium citrate and phosphates. Disintegrants such as starch and lubricants such as magnesium stearate, sodium lauryl sulfate and talc are commonly used in tablets. Convenient diluents for oral administration in capsule form are lactose and high molecular weight polyethylene glycols. When aqueous suspensions are required for oral use, the active ingredient is combined with emulsifying and suspending agents. Certain sweeteners and/or flavorings can be added if desired. For parenteral administration for intramuscular, intraperitoneal, subcutaneous and intravenous use, a sterile solution of the active ingredient is usually prepared and the pH suitably adjusted and buffered. For intravenous use, the total concentration of the solution must be adjusted to make the formulation isotonic. The dosage to be used in humans is ultimately determined by the prescribing physician, but the compound of the present invention or its salt and β-
The daily dosage ratio with the lactam antibiotic is about 1:5 to 5:1, preferably 1:1. The daily oral dosage of each ingredient is usually about 10 mg/kg body weight.
and the daily parenteral dosage of each component usually ranges from about 10 to about 100 mg per kilogram of body weight. However, this number is just an example;
In some cases it may be necessary to use doses outside this range. The invention can be used industrially.

Claims (1)

[Claims] 1 formula [R ⁰ is hydrogen, a salt-forming group or a physiologically hydrolyzable ester-forming group] or a pharmaceutically acceptable salt of the acid or a physiologically hydrolyzable ester of the acid. 2 R ⁰ is phenacyl, acetoxymethyl, pivaloyloxymethyl, α-acetoxyethyl, α
- from acetoxybenzyl, α-hyvaloyloxyethyl, 3-phthalidyl, 5-indanyl, methoxymethyl, benzoyloxymethyl, α-ethylbutyryloxymethyl, propionyloxymethyl, valeryloxymethyl, and isobutyryloxymethyl The ester according to claim 1, which is selected from the group consisting of: 3 formulas [R ⁰ is hydrogen, a salt-forming group, or a physiologically hydrolyzable ester-forming group] In a method for producing an acid, a pharmaceutically acceptable salt of the acid, or a physiologically hydrolyzable ester of the acid, ; a The following formula (A) (wherein R ¹ is benzyl or substituted benzyl) catalytic hydrogenation of the benzyl or substituted benzyl ester, b oxidizing the hydrogenated product to produce the desired acid or substituted benzyl ester in which R ⁰ is hydrogen; and in the preparation of an ester of the target compound in which c R ⁰ is a physiologically hydrolyzable ester-forming group, the above-mentioned acid or its salt is esterified to form a readily hydrolyzable acid. A method consisting of continuous steps to obtain an ester. 4. The method according to claim 3, wherein hydrogenation is carried out using a palladium catalyst. 5. The method according to claim 3, wherein the oxidation is carried out using an alkali metal salt of permanganate or an organic peracid. 6 formula [R ⁰ is hydrogen, a salt-forming group, or a physiologically hydrolyzable ester-forming group] In a method for producing an acid having the following formula, a pharmaceutically acceptable salt of the acid, or a physiologically hydrolyzable ester of the acid, a by KMnO ₄ D, H ₂ O ₂ or other similar peroxides or peracids; Oxidizing the ester with the formula b reacting the ester sulfoxide with a metal in an acid, such as zinc in glacial vinegar, to produce an acid or a salt thereof of the target compound in which R ⁰ is hydrogen, and In the production of an ester of a target compound in which c R ⁰ is a physiologically hydrolyzable ester-forming group, the above-mentioned acid or its salt is esterified to obtain an easily hydrolyzable ester of the acid, using a continuous process. How to become. 7 Formula as the active ingredient in the presence of a carrier or diluent A β-lactamase-producing bacterium containing an acid or a pharmaceutically acceptable salt of the acid.
Pharmaceutical composition for enhancing the effect of lactam antibiotics. 8 Formula as active ingredient [However, R ⁰ is phenacyl, acetoxymethyl,
pivaloyloxymethyl, α-acetoxyethyl, α-acetoxybenzyl, α-hyvaloyloxyethyl, 3-phthalidyl, 5-indanyl,
Methoxymethyl, benzoyloxymethyl, α-
ethylbutyryloxymethyl, propionyloxymethyl, valeryloxymethyl, and isobutyryloxymethyl. 9 Formula as the active ingredient in the presence of a carrier or diluent An antibacterial composition comprising a β-lactam antibiotic and a β-lactamase-inhibiting free acid or a pharmaceutically acceptable salt of the acid.