SU309520A1 - METHOD OF OBTAINING -) - a-AMINO-p- - Google Patents

Description

The invention relates to a process for the preparation of isomers of penicillanic acid derivatives.

In the literature, there is a known method for producing a-amino-a- (hydroxyphenylacetamio) penicillanic acid of the general formula

/ S y /

T SNSOS - CH - CH С (СНз) 2 Ш2 OC-NSNSNOH

The method is that 6-aminopenicillanic acid or its salt is reacted with a functional derivative of a-amino- / g-oxyphenylacetic acid. Products are isolated in a known manner. According to this method, three isomers of the above mentioned penicillin and derivative derivatives were obtained - o-, m- and pisomer as a mixture of two optically active epimers (dl). Moreover, of these three isomers, the m- and l-isomers have the highest activity compared to the activity of standard polyvalent penicillin, D- (-) -a-aminobenzylpenilin.

Further studies have shown that of these two isomers (p- and m-), only levorotatory epimers p-iso, the measure shows an activity almost 2 times greater than the activity of standard D - (-) - a-aminopenicillin, and 3 times as large than the activity of levovrash, ayuschey epimer l -isomer. Thus, the proposed method allows to obtain epimers with new properties of a wider spectrum of action compared with the properties of known compounds.

The proposed method for the preparation of 6 - (-) - amino-p-hydroxyphenylacetamido - penicillanic acid, based on a known reaction, consists in the fact that 6-aminopenicyl acid is reacted with the (-) - isomer of carboxylic acid of the general formula

e X is the amino group

a benzyloxycarbonylamino group, an azido group, an amino group protected by a proton, a Y is a hydroxy group, a halo, a residue of the anhydride of this acid or another acid,

R is hydrogen, benzyl, benzyloxycarbonyl.

Products are isolated in a known manner. The resulting compound can be isolated by any method commonly used for the isolation of aminopenicillins. It can be obtained in free form or BD as salt. Since the penicillin molecule contains both basic and acidic properties, two types of salts are obtained. Acid salts, among which some are moderately soluble in water and can therefore be used for isolation, include salts of inorganic acids, such as hydrochloric, phosphoric, or thiocyanic acid, as well as salts of strong organic acids, such as naphthalene-p-sulfonic acid. The main salts include alkali and alkaline earth metal salts, ammonium salts and non-toxic amines. All of these compounds can be either anhydrous or containing water. They may be amorphous or crystalline, but crystalline forms are preferred as more stable. At any stage of the process, products can be cleaned to remove traces of high-molecular weight allergenic impurities. The resulting p. Nicillins can be used in a mixture with appropriate pharmaceutical carriers in various medical dosage forms. In addition, penicillins can be used in various synergistic combinations with known penicillin-resistant penicillins, for example methicillin, cloxacillin, diclo, xacillin, flucloxacillin and nafcillin. Example 1. Separation of ss-benzyloxycarbonylamino - “- hydroxyphenylacetic acid. The solution of a-benzyl, xycarbopylamino- / y-o, xyphenylacetic acid (224 g) and trigrada quinop (285 g) in boiling ethanol (2.5 l) is cooled, the precipitated crystals are collected and recrystallized twice from ethyl alcohol. It is obtained with a yield of 76o / o quinone salt of levogyrate acid, ajg - 158.5 ° (C, 1 in MeOH). After treating the quinone salt (68 g) with dilute sodium hydroxide, removing the quinone by ether extraction and acidifying the aqueous solution, a levorotating acid is obtained, which is recrystallized twice from an aqueous solution of ethyl alcohol. This gives 28 g (85%) of (-) - a-benzyloxycar5 nonylamino-p-hydroxyphenylacetic acid with m.p. 159-161 ° C, 0 ° (C, 1 in MeOH). Found,%: C 64.1; H 5.2; N 4.7. C24H2lN07 Calculated,%: C 63.9; H 4.9; N 4.7. rope, which is treated with an aqueous solution of caustic soda; the quinone is removed by ether extraction. Upon acidification of the aqueous layer, a crude degrading acid (108 g) is obtained, which is collected, dried and treated with ephedrine (63 g) in boiling ethyl alcohol (450 ml). When the solution is cooled, the prodrug-reducing ephedrine salt is recovered, which is collected and recrystallized from ethyl alcohol. Output Ills, afo -f46.8 ° (C, 1 in NU). The acid is separated from this salt in the usual way, to obtain (- | -) - a-benzyloxycarbonylamino-p-hydroxyphenylacetic acid (960/0), which, after recrystallization from a 50% aqueous solution of ethyl alcohol, has a m.p. 158-16PC,: -f 120.2 ° (C, 1 in MeOH). Found, C 64.1; H 5.3; N 4.8. C24H2 | NO7. Calculated,%: C 63.9; H 4.9; N 4.7. Example 2. Preparation of two e.mers of os-benzyloxycarbonylamino-p-hydroxybenzyl-nicillin. a) Ethyl chloroformate (6.3 ml) was added dropwise at a temperature of -5 ° C to a vigorously stirred solution of (-) - a-benzyloxy "arbonylamino-p-hydroxyphenylacetic acid (20 g) and 2,6-lutidine (8.3 ml) in pure dry acetone (140 ml). The mixture is maintained at a temperature of from -5 to + 5 ° C for 30 minutes to completely form ethoxyuretic anhydride. Then, an ice solution of 6-aminopenicillanic acid (14.3 g) and 2,6-lutidine (10.5 ml) in water (140 ml) is quickly added, and the mixture is stirred at a temperature of -5 ° C for 30 minutes then another 1 hour at room temperature. The solution is concentrated under reduced pressure and temperature, the acetone is distilled off, then the aqueous concentrate is taken up with methyl isobutyl ketone (100 ml) and the pH is adjusted to 2 by adding Dilute hydrochloric acid. The layers are separated, and the aqueous phase is extracted with additional methyl isobutyl ketone (2 x 50 ml). The organic extracts are washed with water, then neutralized with an aqueous solution of sodium bicarbonate to pH 7. The separation of the layers and evaporation of the neutral aqueous solution at low temperature and pressure gives the solid sodium salt of 6 - (-) - a-benzyloxycarbonylamino-p-hydroxyphenylacetamide-penicillanic acid (32 g, 92%). b) By repeating the procedure described above, but using (+) - isomer a-benzyloxycarbonylamino-p-hydroxyphenylacetic acid, 6 - (+) - abenzyloxycarbonylamino-p-hydroxyphenylacetamido) -penicillanic acid sodium salt is obtained, yield 90%, Example 3. Preparation of 6 - (-) - a-amine-hydroxyphenylacetamido-.-Coenicillanic acid ..

A suspension of 50 / o-palladium catalyst - calcium carbonate (40 g) in water (150 ml) is shaken by passing hydrogen for 1 hour, a solution of sodium salt is added - 6 - (-) a-benzyloxycarbonylamino - hydroxyphenylacetamido - penicillanic acid (16 g) in water (50 ml). The mixture was shaken in hydrogen for 40 minutes, then the catalyst was filtered through kieselguhr. pillow and washed with water (3X50 ml). The combined filtrate and washings were adjusted to pH 1.9 and washed with methyl isobutyl ketone (3 x 50 ml). The aqueous phase is adjusted to pH 5.1 and concentrated at low temperature and pressure to a volume of about 75 ml. The colorless crystalline solid that separated was isolated, washed with a small amount of water, acetone and ether. After drying in a vacuum desiccator, 6.3 g (yield 56%) of aminopenicillin is obtained, which is analyzed as trihydrate.

Found, o / o: C 46.0; H 6.5; N 10.3; S 7.4.

C, 6Hi9N305S-3H20

Calculated,%: C 45.8; H 6.0; N 10.1; S 7.6.

When stored in humid air, the sample absorbs water and turns into tetrahydrate + 24Q, 5 ° (C, 0.1 in H2O).

Found,%: C 44.1; H 6.1; N 10.0; S 7.6; NaO 16.3.

C, bN19Nz055-4N20

Calculated,%: C 43.9; H 6.2; N 9.6; S 7.4; HgO 16.4.

Example 4. 6 - (+) - a-amI | But - “- hydroxyphenyl acetamido-penicillanic acid.

Sodium salt of 6 - (+) - a-benzyloxycarbonylamino - / I-hydroxyphenylacetamido - tsenicillanic acid is hydrogenated in the same way as the epimer in Example 3. The crystalline product (6.1 g, 54 ° / o) after drying in a vacuum desiccator and leveling - air humidity implies -f 154.6 ° (C, 0.1 in HgO) and in the analysis it is tetrahydrate.

Found,%: C 43.7; H 6.5; N 9.3; S 7.6; HgO 15.8.

C, 6H, 9N305S-4H20

Calculated / a: C 43.9; H 6.2; N 9.6; S 7.3; HgO 16.4.

Example 5. The separation of a-benzyloxycarbonylamino-w-hydroxyphenylacetic acid.

The racemic l-hydroxy acid is dissolved with quinidine in boiling ethyl alcohol (1.875 l1 mol}. When cooled, the quinidine salt of the (-) - isomer crystallizes. The crystals are collected, boiled in ethyl alcohol (1,3), the suspension is cooled and filtered. From the salts release free

an acid substantially similar to that described in Example 1 for the p-hydroxy isomer a. By recrystallization of I.H. chloroform or toluene, a yield of 79% (-) - a-benzyloxycarbonylamino-lg-hydroxyphenylacetic acid, -110.8 ° (C, 1 vMeOH) is obtained.

The ethanol filtrate and washings from the indicated crystallization of the quinidine salt are evaporated in vacuo, the remaining syrup

crystallized by trituration with ether. Upon recrystallization from isopropanol, the quinidine salt of (+) -acids is obtained, from which the free acid is isolated in the usual way. When recrystallized from

chloroform or toluene is obtained with a yield of 73% (-f-) -a-benzloxycarbonylamino-llyoxyphenylacetic acid.

 -f 110.5 ° (C, 1 in MeOH).

Example 6. Preparation of two a-benzyloxycarbonylamino-m-hydroxybenzylpenicillin epimers.

These substances are obtained as in example 2.

From (-) -а-benzyloxycarbonylamino-g-hydroxyphenylacetic acid, sodium salt of 6 - (-) - a-benzyloxy-polycarbonylamino-lg-hydroxyphenylacetamido - penicillanic acid is obtained. 90% yield.

From (- | -) -а-benzyloxycarbonylamino-l4-hydroxyphenylacetic acid, sodium salt of 6- (+) - a-benzyloxycarbonylamino-UI - hydroxyphenylacetamido - penicillanic acid is obtained. Yield 90%.

Example 7. Getting 6 - (-) - a-amino. Ioxiphenylacetamido-penicillanic acid.

Catalytic hydrogenation is carried out with 6 - (-) - c-benzyloxycarbonylamino-m-hydroxyphenylacetamido-peptide of nicillanic acid according to the procedure described in Example 3. The aqueous solution of the reduced penicillium is brought to pH 5.5 and concentrated etc. At low temperatures and pressure .. The output of hydrogenated aminopenicillin 50E / o, -f-239 ° (C0.2 in NZO). Karl Fischer test shows that crystals contain 15% water.

Example 8. Preparation of 6 - (+) - a-amino-joxyphenylacetamido-penicillanic acid. The experiment was carried out according to the method described in

Example 7, but the raw material used is the sodium ionic salt of 6- (+) -a-benzyloxycarbonylamino-m-hydroxyphenylacetamido-penicillanic acid. The output of acid bo / o. Crystalline aminopenicilli .n contains 15% water, 4-135 ° (C0.2 in NgO).

Example 9. Getting 6 - (- 4 -) - a-amino. And - (/ g-hydroxyphenyl) - acetamido - penicillanic acid.

a) Suspension of (-) - a-benzyloxycarbonylamino-w- (p-hydroxyphenyl) acetic acid (21.8 g) in water (180 liter) is treated with a dilute sodium hydroxide solution. Get a clear solution with a pH of 8.7. Next, 5% palladium on a calcium carbonate catalyst (2.2 g) is added, the mixture is shaken in hydrogen at atmospheric pressure until the absorption of hydrogen ceases. The catalyst is filtered through a kieselgur pillow and washed with water. The combined filtrate and washings were adjusted to pH 5.0 and the solution was evaporated in vacuo to a small volume. The crude product is separated as a gelatinous precipitate, which is dissolved by heating the mixture to a boil. When the solution is cooled, colorless crystalline (-) - a-amino-a - ("hydroxyphenyl) acetic acid is released, which is isolated, washed with a small amount of cold water and existed in vacuum over phosphoric anhydride. The output of 10.1 g (83%), so pl. 225-226 ° С, and -108 ° (С, 1 in НгО).

Found, in o / o: C 57.2; H 5.4; N 8.3. CsHgNOa

Calculated,%: C 57.5; H 5.4; N 8.4.

Normal magnetic resonance D2O + -4-NaOD: the multiplet is centered on t 3.15 (4H, aromatic), singlet t 5.8 (-CH).

b) A solution of sodium hydroxide in methyl alcohol (60 ml, 0.324 N, 0.01595 mol) is added to (-) - a-amino-o- (l-hydroxyphenyl) - acetic acid (3.34 g, 0, 02 mol) and washed with methanol (20 ml). When the suspension is heated to reflux almost to the boiling point, a clear solution is obtained from which the crystalline sodium salt of the amino acid is separated.

While moving, to the boiling suspension, the p. Solution of methyl acetoacetate Ta (2.4 ml, 0.022 mol) in methyl alcohol (20 ml) is added over 10 minutes while boiling for another 20 minutes. A clear solution is obtained which is boiled for another 20 minutes. Methyl alcohol is distilled off (bath temperature 130 ° C) and a small amount of dry toluene is immediately added. After adding 90 ml of toluene, a white crystalline substance is released. The process continues until the distillation temperature reaches 100 ° C and only 170 ml of toluene has not been introduced. The suspension is left overnight at 5 ° C, then the product is isolated, washed with dry toluene and dried under vacuum at 40 ° C over phosphoric anhydride. The yield of sodium - (-) - Nmethoxycarbonylpropen-2-nl) -a-amino - a- (coxiphenyl) -acetate 4.5 g (95 ° / o) Found: C 54.35; H 4.9; N 4.9.

CiaHnNOsNa BychiCv Eno,%: C 54.4; H 4.9; N 4.9.

Infrastructure (nuzhol) 3300 cm-i (N-H-), 1655 CM-i (C O-), 1560 cl-1 (COO-).

c) A stirred suspension of 6-aminopenicillanic acid (2.16 g, 0.01 mol) in

water (25 ml) is adjusted to pH 7.8 by adding 10%. aqueous solution of caustic soda. It should be ensured that the pH does not exceed 8.0. Dry acetone (40 ml) is then introduced and the solution is cooled to -10 ° C.

Dry acetone (25 ml) is stirred and maintained at a temperature of from -10 to -8 ° C while adding chloroholic acid ethyl ester (1.0 ml, 0.0104 mol). Then lo / 0-th n-methoxycarbonylpropen-2-yl) -a-amino-a - (/ g-hydroxyphenyl) -acetate (2.87 g, 0.01 mol) is added, which is washed in cold, dry acetone (15 ml ). The mixture is moved at -10 ° C for 30 minutes, then it is poured into the sodium-6-aminopenicillinate solution described above and washed in cold dry acetone (20 ml). The mixture was stirred for 20 minutes at -10 ° C, the acetone was distilled off in vacuo at room temperature, and isobutyl methyl ketone (40 ml) was added to the remaining aqueous solution. Next, the mixture is cooled to 5 ° C, stirred vigorously and the pH is adjusted to 0.9 with concentrated hydrochloric acid.

The mixture was maintained at a pH of 0.9 and a temperature of 5-10 ° C for 75 minutes, then the layers were separated. The aqueous layer was washed with isobutyl methyl ketone (10 ml), the residue of the solvent was removed in vacuo. Water solution,

cooled to 5 ° C and adjusted to pH 5.2 U / o-s sodium hydroxide solution. The resulting suspension was stirred at 50 ° C for 5 minutes, then concentrated in vacuo at 20 ° C to Vs of the same volume. White crystalline 6 - (-) - a-amino "- (-oxiphenyl) - acetamido - penicillanic acid is isolated, washed with water (20 ml) and dried in air at 40 ° C.

Infrared spectroscopy and spectroscopy using normal magnetic resonance and fluorescent paper chromatography show that the resulting product (2.8 g) is similar to the product in Example 3.

The intermediate step described in paragraph b can also be carried out as follows, d) A stirred suspension of 6-aminopenicillic acid (4.32 g, 0.02 mol) in water (50 ml) is adjusted to a pH of 7.8, carefully add 10% potassium hydroxide solution. Acetone (80 ml) is then introduced and the suspension is cooled to -10 ° C. Dry acetone (50 ml)

stirred and maintained at a temperature of from -1 to -8 ° C, while entering the ethyl ester of chlorogolic acid (2.0 ml, 0.0208 mol). Next, add 1% n-methylmorpholine in acetone (3 ml), and then sodium amino-a- (p-hydroxyphenyl) acetate (5.74 g, 0.02 mole), which is washed with cold dry acetone (30 ml The mixture was stirred at -10 ° C for 30 minutes until the mixed anhydride was completely formed in the solution, then cooled to -25 ° C and quickly filtered through acid-washed kieselgur. The filter mixture was washed with dry cold acetone (20 ml, and the filtrate with; wash water is added to a cold stirred aqueous acetone solution of the above-described 6-aminopenicillium potassium.

The clear solution is stirred for 30 minutes, then without external cooling for another 30 minutes, then evaporated to dryness under reduced pressure and a temperature below 20 ° C. The gummy residue is dried in vacuo over phosphoric anhydride. The weight of the crude product is 9.7 g. Most of the product (9.5 g) is treated with dry methyl alcohol (70 ml in which most of the product dissolves. A small amount (0.54 g) of the undissolved 6-a. Minepenicillanic acid is removed The yellow filtrate is stirred for 5 minutes with activated carbon (2.3 g), then the mixture is filtered again. The filtrate is diluted with dry isopropyl alcohol (250 ml., causes crystallization and the mixture is left overnight at 5 ° C. The resulting white the solid product is filtered, washed with dry ether, dried in a vacuum . IU over phosphorus pentoxide to give 6 hemihydrate - (-) - N (1-methoxycarbonylpropyl-2-yl) - it - amino- (p-hydroxyphenyl) acetamido -penitsillat potassium.

Found,%: C 49.25; H 5.2; N 8.3; S 6.35; K 7.6.

C2, H24N3O7SK-0,5 NgO

Calculated, o / o: C 49.4; H 4.9; N 8.25; S 6.25; K 7.65.

From the aqueous solution of this salt, acidified to pH 2, methyl acetoacetate and 6 - (-) - a-amino-a - (/ g-hydroxyphenyl) acetamido-penicillanic acid are quickly isolated.

Example 10. Preparation of 6- (-) -a-amino hydrochloride trihydrate (p-hydroxyphenylacetamido-penicillanic acid).

Finely divided 6 - (-) - amino- (p-hydroxyphenyl) acetamido - penicillanic acid trihydrate (2 g suspended in water (5 ml) and 5N hydrochloric acid (2.5 ml) added. The mixture is shaken to dissolve most of the solid material and quickly filter. The filtrate is seeded with crystals and left at 5 ° C for 30 minutes until the beginning of the precipitate of hexagonal crystals. The crystals are filtered, washed with a little cold water and dried in an oven at 35-40 ° C Get trihydrate hydrochloride 6 - (-) a-amino-p-0 | Xyphenylacetamido - penitsi Lanovoi acid (1.6 g) which turns brown with

90 ° C, then dehydrated at 216-218 ° C (with decomposition). Hydroxylamine phosphate 100%. Mol weight. 496.0. The content of NgO 11.50 / 0.

Found,%: C 42.1; H 5.7; N 9.1; S 7.2; C1 7.9.

Calculated for Ci6H2oN3O5Cl-3H2O,%: C, 42.2; H 5.7; N 9.2; S 7.0; From 7.8.

Example 11. Preparation of 6 - (-) - a-amino-g-hydroxyphenylacetamido - penicillanic acid disulfate.

A suspension of finely divided 6- (-) -a-amino-p-hydroxyphenylacetamido-penicillanic acid trihydrate (2 g) in water (200 ml) is prepared and 5N sulfuric acid (2.5 ml) is added. The mixture is stirred at room temperature 5 min, then filtered and evaporated to a small volume (about

50 ml} at low temperature and pressure. The white matter that crystallized out of the solution was filtered and washed with a small amount of cold water and dried in an oven at a temperature of 35-40 ° C. Get

while the disulfate is 6 - (-) - a-amino- / g-hydroxyphenylacetamido-penicillanic acid (1.2 g). T. pl. (turns brown) 210 ° C, decomposition. 230- PC. Hydroxyl.minova sample 227%, H2O content 1.1 o / o.

Found,%: With 46.1; H 4.9; N 10.1; S 11.8.

Calculated for CssHMNeSsOu, / o: C, 46.4; H 4.9; N 10.1; S 11.6.

Example 12. Obtaining (-) - a-benzyloxycarbamido-a- (/ 1 - benzylcarbonyloxyphenyl) -acetic acid.

a) To a strongly stirred ice solution of (-) - a-benzyloxycarbonamide-a - (coxyphenyl) -acetic acid (15 g) and caustic soda (7 g) in water (1500 ml) add chloryl ester benzyl ester (13 g The solution is stirred for 2 hours, then extracted with ether. A small amount of solid precipitate is filtered and discarded. An air stream is passed through the separated aqueous phase to remove the dissolved ester. When acid is added, excess resin precipitates to excess hydrochloric acid. about him Benzene is poured into the stack (200 ml}. When heated in a reflux vessel, the resin dissolves and azeotropic water is removed (Dean-Stark unit). When cooled

get a solution of (-) - a-benzyloxycarbamido-a- (p - benzyloxycarbonyl xyphenyl) acetic acid in benzene in the form of a white crystalline solid (14 g), so pl. 94-95 ° C.

The analysis shows that the compound contains a crystallization benzene molecule. Found,%: C 69.8; H 5.3; N 2.8.

Calculated,%: C 70.2; H 5.3; N 2.7; CDCU, ppm: 10.34 (S, 1H, NH); 7.25 (t, Ar, 2OH); 5.35 (s, 1H, CH); 5.25 (S, 2H, CHg) and 5.05 (S, 2H, SNg).

b) Preparation of (-) - N-carboxy - (rt-benzyloxycarbonyloxyphenyl) -glycyl anhydride.

Thionyl chloride is added to the benzene solvate of D-a-benzyloxy-carbamido-os - ("- benzyloxycarbo-yloxyphenyl) acetic acid (4.35 g) and the solution is refluxed for 2 hours. Excess thionyl chloride is removed under reduced pressure, traces of it are removed by distillation together with dry benzene under reduced pressure, and the residue is kept in a vacuum for 30 hours. Dry ether (15 ml) is added to the resinous precipitate, and a clear solution is obtained in which (-) - a-carboxy- (d-benzyloxycarbonyloxyphenyl) -glycyl anhydride precipitates in 5-10 minutes as a white crystalline solid (1, 33 g) T. pl. 118-120 ° C.

Found,%: C 62.3; H 4.1; N 4.3.

CirHisNOe

Calculated, o / o: C 62.4; H 4.0; 4.3.

c) Preparation of 6 - (-) - a-amino-a- (p-benzyl: xycarbonyloxyphenyl) - acetamido - penicillanic acid.

A suspension of 6-aminopeNicillanic acid (0.66 g) in water (100 ml) was adjusted to pH 5.5 and, with vigorous stirring, added to the suspension at a temperature of 5 ° C in portions every 5 min. (-) - a-carboxy - (p-benzyloxycarbonyloxyphenyl) -glycyl anhydride (0.99 g). It should be ensured that the pH value was an av. 5.5. Stir vigorously to continue e, f for 2 hours at a temperature of 5-10 ° C. The mixture is then filtered through diatomaceous earth, the filtrate is concentrated to 25 ml. It precipitates in the form of a white solid substance 6 - (-) - a-amino-a- (d-benzyloxycarbo-Nyloxyphenyl) -acetamido-penicillic acid (0.09 g).

Analysis by paper chromatography shows that the product contains approximately 5% impurity of 6-a. Aminopenicillanic acid. In the system butanol-ethanol - water 6 - (-) - aa.mino-a - (/ g - benzyloxycarbonyloxyphenyl) -acetamido - penicillanic acid Rf 0.47; 6-amyiopenicillanic acid Rf 0.13.

The product inhibits E. Coli 2 at a concentration of 5 µg / ml and Staiph Oxford at a concentration of 0.1 µg 1 ml.

Hydrolysis of the O-benzyloxycarbonyl group.

The aforementioned penicillin is dissolved in phosphate buffer pH 9 and maintained at this pH. Portions are selected and periodically analyzed by paper chromatography. After three days, at room temperature, the conversion to 6 - (-) - a-amino- (p-hydroxyphenyl) -acetamide-pencylitanic acid occurs almost completely.

Example 13. Preparation of 6- (-) - a-a. Amino- (p-hydroxyphenyl) - acetamido - penicillanic acid.

Phosgene is passed through a suspension of (-) - a-amn} 10-a, - (p-hydroxyphenyl) acetic acid (4 g) in dioxane (150 ml) at a temperature of 50-60 ° C until a clear solution is obtained. Excess phosgene is displaced with nitrogen and the solution is filtered. Dioxane is removed from the filtrate at a temperature of 50 ° C and under reduced pressure; oil remains,

which hardens when kept in vacuum over phosphoric anhydride. The solid is dissolved in boiling acetate ether (50 ml); a small amount of tar remains. Activated charcoal is then added and the mixture is transferred to cool to room temperature. After filtration through kieselguhr, the filtrate is slowly poured into petroleum ether stirred at 40-60 ° C.

(400 ml) for vysalivaniya (-) - cc-carboxy (p-oxyphenyl) -glycyl anhydride in the form of a solid white crystalline substance (3.1 g), so pl. 260 ° C (with decomposition). Found With 56.1; H 3.7; N 7.1.

C9HrN04

Calculated,%: C 56.0; H 3.65; N 7.25.

A suspension of 6-aminopenicillanic acid (1.5 g) in water (100 ml) is adjusted to pH 5.6 by adding caustic soda. To the stirred suspension at 5 ° C is added (-) - a-carboxy- (p-hydroxyphenyl) - glycyl anhydride (1.5 g) in portions every

5 minutes. The mixture is stirred at a temperature of 5-10 ° C for 1 hour and then another 1 hour at room temperature. This causes a strong foaming, which is inhibited by 1-2 drops of n-octyl alcohol. Further

the mixture is filtered through kieselguhr and the pH of the filtrate is adjusted to 5.0. When the concentration of the precipitate does not fall out, so the solution is evaporated to dryness. A nearly white solid is obtained (1.7 g). A paper chromatography analysis shows that the product contains 6-aminopenicillanic acid and the required 6 - (-) - a-amino-a- (p-hydroxyphenyl) -acetamido-pecicillanic acid in approximately equal amounts.

Example 14. The reaction of 6-aminopenicillanic acid with (-) - a-amino-a- (d-hydroxyphenyl) -chloroacetyl hydrochloride. Prepare a suspension of (-) - a-amino- (x- (g-hydroxyphenyl) -acetic acid (1.67 g) in chloroacetyl (50 ml) and cool to 0 ° C, while adding finely powdered phosphoric anhydride (4 , 16 g) with stirring. Then dimethylformamide is added