DK144177B - METHOD OF ANALOGUE FOR THE PREPARATION OF 7- (ALFASULFOACETAMIDO) -3- (HETEROCYCLYL-THIOMETHYL) -CEP-3-EM-4-CARBOXYLIC ACID DERIVATIVES OR PHARMACEUTICAL ACCEPTABLE SALTS. - Google Patents

Description

i 144177 o

The invention relates to an analogous process for the preparation of novel 7- (α-sulfoacetamido) -3-freterocyclyl-·-thiomethyl) -ceph-3-em-4-carboxylic acid derivatives of the general formula 5 g

r1-chconh _S

I I 2 SO 3 H L_N Λ — CH 2 SR (i)

10 COOH

wherein R 2 represents a hydrogen atom, phenyl or thienyl, and 2 R represents a pyridyl, pyrimidyl, imidazolyl, thiazolyl, thiadiazolyl, or tetrazolyl group which may be substituted by methyl, amino, thio, mercapto, methylthio, phenyl and / or acetamido, or pharmaceutically acceptable salts thereof.

From the specification of Danish patent application No. 606/71 20 antimicrobially active cephalosporanoic acid derivatives of the formula are known.

S

R-CHCONH —_N

25 SO-, Μ -N -CH2X

J o COOM 'where R may mean hydrogen, alkyl or phenyl, M and M' e.g.

30 may mean hydrogen or an alkali metal and X means -COCH 3 or

35 -c- I

ISLAND

ISLAND

2 144177

However, the compounds of the present process have a better effect against e.g. Ps. aeruginosa and E. Coli than these known compounds, as evidenced by comparative experiments described in the experimental part of the specification.

It has been found that the conversion of the 7-position acyl group into cephalosporin C, i.e. a 5-amino-5-carboxyvaleryl group, to an α-sulfoacyl group, provides a compound effective against Pseudomonas aeruginosa against which known cephalosporins such as cephalothin or cephaloridine are ineffective. It has also been found that the compound has a wider antimicrobial spectrum than cephalothin and cephaloridine. Furthermore, it has been found that effects as mentioned above are further enhanced by additional substitution of the acetoxymethyl group at the 3-position with a heterocyclic thiomethyl group.

The cephalosporins of formula (I) can be prepared by converting the 5-amino-5-carboxyvaleryl group at the 7-position in cephalosporin C to the desired α-sulfoacyl group and by converting the acetoxymethyl group at 3-position to the heterocyclic thiomethyl group. The conversion can be performed first in the 3 or 7 position as desired, followed by conversion in the second position.

The process of the invention is characterized in that (a) a sulfocephalosporin of the general formula

S

R1-CHCONH —- ζ ^ 30 lo3H J-AN ^ L-CH2OjCH3 <IX> T °

COOH

wherein R · is as defined above, or a salt and the like thereof, is reacted with a heterocyclic compound of formula 2

ISLAND

R2-SH (III) wherein R is as defined above or a salt thereof, or 5 (b) an amino-cephalosporanoic acid derivative of the general formula

S

m2-I- {10 ._1 Å-CH2 SR2 (iv)

COOH

Wherein R is as defined above or a salt thereof, or a readily cleavable ester thereof, is reacted with a compound of general formula 20

-CHC00H R

I (V)

S0, H

? Wherein the above meaning, or a functional derivative of the carboxyl group, is cleaved, whereupon an ester group optionally present in the reaction product is cleaved and if desired a converted salt is converted to the free acid and / or the free acid is converted to a pharmaceutically acceptable salt thereof .

The two variants of the method are described below.

(a) Method of performing the conversion at the 3 position after prior conversion at the 7 position.

144177

ISLAND

4

The sulfocephalosporin compound (II) used as a starting material can be obtained e.g. by condensation of α-sulfo-carboxylic acids or functional derivatives of the carboxyl group thereof with 7-aminocephalosporanoic acids or easily removable ester derivatives thereof and, when a readily removable ester group is used in the previous step, subsequent removal of the ester group (Belgian patent no. 762,725).

In these sulfocephalosporin compounds (II), the carboxyl group at the 4-position and / or the sulfo group on the side chain 10 thereof can form a salt, e.g. sodium, potassium, magnesium, calcium, aluminum and triethylamine, as long as no detrimental effect is produced in the reaction of the present invention. In some cases, the carboxyl group in 4- "position may be a group which is easily removable, e.g.

A benzyloxycarbonyl, β-methylsulfonylethyloxycarbonyl, benzhydryloxycarbonyl or trimethylsilyloxycarbonyl group.

As mentioned, 2 R of the heterocyclic thiol compound (III) may be pyridyl, pyrimidyl, imidazolyl, thiazolyl, thiadiazolyl or tetrazolyl. These rings may have as substituents e.g. a methyl group, an amino or thio group. As an alternative, they may be partially reduced, e.g. in dihydroimidazole. The nitrogen atom of the heterocyclic thiol compound (III) may be quaternary, such as in N-methylpyridinium.

The reaction between the sulfocephalosporin compound (II) and the heterocyclic thiol compound (III) is usually carried out in a suitable solvent. Eg. For example, acetone, dioxane, chloroform, dimethylsulfoxide, methylene chloride, tetrahydrofuran, ether, ethyl acetate ester, nitrobenzene, dimethylformamide, methanol or ethanol may be used. Generally, other organic solvents that will not affect the reaction as well as water can also be used. Among these solvents, especially those with high polarity are preferred. The hydrophilic solvents can be mixed with 35 water according to the application. The reaction is preferably carried out

ISLAND

5 144177 at a neutral or slightly acidic pH. It can e.g. preferably is carried out in the presence of an alkali metal hydroxide, an alkali metal carbonate, an alkali metal hydrogen carbonate, a trialkylamine or pyridine. The reaction temperature varies depending on the nature of the compounds used as starting materials. Generally, however, the reaction is carried out at a temperature ranging from room temperature to 100 ° C, especially from 40 to 60 ° C. In most cases, the reaction is continued for approx. lr-24 hours or more. The heterocyclic thiol compound (III) is preferably used at a ratio of from approx. 1 to approx. 10 moles per mole of sulfocephalosporin compound (II).

(B). Procedure for Performing the 7-Position Conversion After Prior Conversion at the 3-Position · The aminocephalosporanoic acid derivative (IV) can be obtained by substituting the 3-position acetoxy group in cephalosporin C with the desired heterocyclic group and then removing the acyl group in 7- position. On the other hand, can it manufacture? By deacylating cephalosporin C and then substituting the 3-20 - position of the resulting 7-aminocephalosporanoic acid.

The conversion of the 3-position acetoxy group into cephalosporin C can be carried out according to the procedure described in e.g. U.S. Patent No. 3,225,038 or No. 3,217,000 or German. U.S. Patent No. 1,871,121, The resulting cephalosporin derivative having the heterocyclic group at the 3-position can be readily converted according to the method described in e.g. Dutch patent no.

64.01.421, British Patent No. 1,041,985 or U.S. Patent No. 3,575,970 to a corresponding aminocephalo-sporanoic acid derivative (IV).

The aminocephalosporanoic acid derivative may also be in the form of e.g. salts and esters, as is also the case with the above-mentioned sulfocephalosporanoic acid (II).

The reaction between the aminocephalosporanoic acid derivative 35 (IV) and the sulfocarboxylic acid (V) or a functional derivative thereof can also be carried out according to a known method.

ISLAND

In cases where the α-sulfocarboxylic acid (V) is used as free acid, it is preferred to carry out the reaction in the presence of a condensing agent. The condensing agent is e.g. an N, Ν'-disubstituted carbodiimide, e.g. Ν, Ν'-dicyclohexyl-5 carbodiimide; an azolid compound, e.g. N, N'-thionyl-imidazole; N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline, phosphorus oxychloride or alkoxyacetylene. As a functional derivative of the acid (V), e.g. carboxylic acid halides, anhydrides, azides and active esters are used. The above acylation reaction advantageously and smoothly takes place in a solvent. As a solvent, a conventional solvent such as water, acetone, tetrahydrofuran, dioxane, acetonitrile, chloroform, dichloromethane, dichlorethylene, pyridine, dimethylaniline, dimethylformamide or dimethylsulfoxide is suitably used. The reaction temperature is not very limited. Generally, however, the reaction is carried out under cooling or at room temperature.

The reaction product can be purified and recovered using the properties of the cephalosporin end product (I) 20 with respect to e.g. phase transfer, concentration, chromatography, crystallization and recrystallization.

Thus, the compound prepared by the process of the invention can be obtained as free acid or as a salt which, if desired, can be transferred to another type 23 salt by a known salt exchange reaction. The salt may be of any type of a pharmaceutically acceptable non-toxic salt. Among useful types of salt are such non-toxic metal salts as the salts of sodium, potassium, calcium and aluminum, such non-toxic amine salts as ammonia or substituted ammonium salts, e.g.

triethylamine, procaine, dibenzylamine, N-benzyl-p-phenethylamine, 1-ephanamine or N, N'-bis (dehydroabiethyl) ethylene diamine; and salts of other amines used heretofore to make salts of btenzylpenicillin.

The compound prepared by the process of the invention contains two acidic groups, namely one 7 144177

ISLAND

sulfo group and a carboxyl group, and depending on the relative acidity of these two groups, it is possible to obtain either an acidic salt or a neutral salt. In the 7-position alpha-sulfoacyl group there are also one or two 5 asymmetric carbon atoms. Using optically active starting materials, e.g. α-sulfocarboxylic acids, or by subjecting the reaction product to a conventional technique, such as recrystallization or chromatography, optically active forms of the subject compound can be obtained.

The cephalosporin compounds obtainable in the above manner have potent antimicrobial effects and are effective in the treatment of various infectious diseases caused by gram-positive and gram-negative bacteria, including the bacterial group Pseudomonas. They may be administered by any route including oral, subcutaneous and intravenous routes in the same manner as known cephalosporin preparations.

While the appropriate dose depends on the type of disease, the recommended dosage for Pseudomonas infections is e.g. from approx. 5 to approx. 500 mg / kg / day and preferably 10-200 20 mg / kg / day in portions for 2-4 administrations per day.

The following examples further illustrate the process of the invention.

Example 1 (1) A mixture of 2.5 ml of 1N NaOH solution and 5 ml of water is cooled with ice and stirred well at 0-5 ° C. With stirring, 680 mg of 7-aminocephalosporanoic acid is added and dissolved well. By itself, 585 mg of α-sulfophenylacetic acid chloride are dissolved in 7 ml of diethyl ether and the solution is added 30 drops dropwise to the previously prepared solution over 15 minutes. The aqueous layer is recovered and adjusted to a pH of 1.5 with 1N HCl followed by extraction with two 15 ml portions of n-butanol. The extracts are washed twice with 5 ml water portions and extracted with a saturated aqueous solution of NaHCO 3. The extract is brought to a pH of 6.5 and washed with ether followed by lyophilization. 385 mg of sodium salt of 7- (α-sulfo-phenylacetamido) -cephalosporanoic acid is obtained.

δ 144177 ο (2) 293 mg of disodium 7- (α-sulfophenylacetamido) -cephalosporanate, 188 mg of 2,5-dithio-1,3,4-thiadiazole and 13 mg of NaHCO 3 are dissolved in 25 ml of water. The solution is adjusted to a pH of 7.0 and heated to 40 ° C for 45 hours.

The reaction mixture is brought to a pH of 2.5 and washed with 25 ml of ethyl acetate. The aqueous layer is then extracted with n-butanol. To the extract is added water and adjusted to a pH of 6.5 with NaHCO 3, thereby transferring the intended product to the aqueous phase. The aqueous phase is lyophilized, then the lyophilized product thus purified chromatographically using a (S) resin column ("Amberlite XAD-2

Rohm & Haas Co., USA, 16 x 1000 mm, eluant: water).

The fractions containing the desired cephalosporin are cooled and then lyophilized to give 120 mg of sodium 7- (α-sulfophenylacetamido) -3- [5 '- (2'-thio-1', 3 ', 4'- thiadiazole) thio] methyl-3-cephem-4-carboxylate.

KBz * -1 IR absorption spectrum γ> max (cm): 1750 (β-lactam, C = O), 1670 (-CONH-), 1600 (-COO-), 1530 (-NC = S), 1285 ( CSNH), 1400, 1360, 1215 (SO2), 1045 (-SO3H).

NMR spectrum (60 MC, D 2 O): 3-4 (2H, C 2 -H 2), 5.1 (1H, S, -CH- in side chain), 5.06 (1H, doublet, J = 5 cps,

Cc-H), 5.62 (1H, doublet, J = 5 cps, C - H), 25 ^ 7.5 (5H, broad S, phenyl-H).

Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 209 P 2

Pseudomonas aeruginosa 10490 20 30

Example 2 Dissolve 0.880 g of disodium 7- (α-sulfoacetamido) cephalosporan (2 x 10 mol) in 50 ml of water. Then, 0.75 g (5 x 10 6 mol) of 2,5-dithio-1,3,4-thiadiazole is added to this solution and mixed well. The mixture is adjusted to a pH of 6.5 by means of a dilute aqueous

ISLAND

And then the reaction is carried out at a temperature which is kept constant at 40 ° C for 8 hours. After completion of the reaction, the reaction mixture is concentrated at room temperature under reduced pressure until the volume of liquid is about 5%. 30 ml. The concentrate is subjected to lyophilization. The resulting product is then purified chromatographically using a resin column ("Amberlite XAD-2", 30 x 1000 mm, eluent: water). The fractions containing the desired cephalosporin are collected and then lyophilized to give 500 mg of sodium 7- (α-sulfoacetamido) -3- [5 '- (21-thio-1', 3 ', 4'-thiadiazol) thio] methyl - 3-cephem-4-carboxylate.

IR V (cm -1): 3400 (OH), 3000 (CH), 2850 (NH), ΠΙαΧ 1745 (β-lactam, C = O), 1665 (-CONH-), 1600 (-COO ~), 15 1460 (-CSNH-), 1400, 1260, 1220 (-SO2 ~), 1120, 1100 (CNH-), 1,045, 1025 (-SO, ~), 700 (CS).

II

p

Minimum inhibitory concentration (γ / ml):

Pseudomonas aeruginosa 10490 10 20

Example 3, 246 g of disodium 7- (a-sulfophenylacetamido) cephalo-sporanate, 0.149 g of 2,4-dithio-pyrimidine and 0.13 g of NaHCO3 are dissolved in 21 ml of water. The mixture is heated to 40 ° C for 41 hours. At the end of the reaction, the reaction mixture is adjusted to a pH of 2.0 with HCl. Then the mixture is treated in the same manner as in Example 2 to give 0.10 g of 7- (α-sulfophenylacetamido) -3- [4 '- (2'-thio-pyrimidyl) -thio] -methyl-3-cephem 4-carboxylic acid.

IR ^ ^ (cm -1): 3380 (OH), 3000, 2900 (CH),

ulclX

1750 (β-lactam, C = 0), 1665 (-CONH-), 1600 (~ C00 ~), 1535 (CN, NC = S), 1470 (NCS), 1370 (SO2), 1290 (NCS), 1230 , 1190 (SO2), 1115, 1045 (-SO3_), 700

Minimum inhibitory concentration (γ / ml): Pseudomonas aeruginosa 10490 10 10 144177 o

Example 4 Dissolve 0.217 g of disodium 7- (α-sulfophenylacetamido) cephalo-sporanate with a thiol compound in 15 ml of water.

The mixture is adjusted to a pH of 6.4 and heated to 5 to 40 ° C for 18 hours. After completion of the reaction, the reaction mixture is subjected to lyophilization and then chromatographically purified using cellulose in powder form (eluent: n-propanol: water: trichloroacetic acid = 75: 25: 1). The fractions containing the desired 10 cephalosporins are collected and concentrated under reduced pressure. Then n-hexane is added to the concentrate to precipitate crystals. The crystals are collected and dissolved in water. After the solution is adjusted to a pH of 6.4, it is again subjected to lyophilization to obtain the intended products.

Several experiments have been carried out in this way using different thiol compounds, viz. 4-amino-2-thiopyrimidine and 4,5-dihydro-2-thioimidazole.

20

Substitution in 3-Kgr-1 position IRV / raax (om-) +, m2 3400 (OH), I76O (β-lactam), 1670 (-CONlf), 25 + CH3 + SN__s / 1610 (C00 ~), 1210 (-SO2-) and 1038 (-30 ^) 3450 OH), 1760 (β-lactam), l665 (-CONH ~), ΌΗ23 \ ν_ | 1615 (000 °), 1210 fflOg-) and 1040 (SOjr) 30__5 __-_____

The minimum inhibitory concentration of the products obtained is given in the following table.

35 144177 11 · E 11 --------- 1 1 '"· -----” -------' 'Ί' ~

Substituent in the 3-position Bacteria ___mix (µg / ml), "" Pseudomonas aeruginosa 5.07 nh2. _: - "- 4

Bacillus subtilis 2.9 s -ch9s-L 0 2 '—'— »-"' 1 "I ·" 11 1 I "1111 11 '1' —..." MM.IHI-I "" * -I— II »-!. ...... · - H '/» S! -CH9S-S I' Bacillus subtilis 1 3.9 10 ^ NJ;

H i I

. · ............ ......... — ... I ................... „.. ., .. ....... IU,, ---—

Example 5 517 mg of disodium [7- (α-sulfophenyllaetamido) cephalosporanate, 525 mg of 2-acetamino-5-mercapto-1,3,4-thiadiazalyl and 1.75 g of KSCN are mixed in 2 ml of water. After adjusting the mixture to a pH of 6.5, it is heated to 60 ° C for 6 hours. The heating mixture is purified by column-20 chromatography using Amberlite XAD-2 resin to obtain 240 mg sodium ~ 7- (α-sulfophenylacetumido) -3- (2'-acetamino-1 ', 3', 4'-thiadiazolylthio) methylceph-3-, em-4-carboxylate.

IR ν + (cin) 1: 1760 (B-lactam), 1670 (-CONH-), ΓΠαΧ 1610 (-CO2-), 1040 (SO3 KMR (D20): 2 (3H, S, - CH3), 5.10 (1H, S, éP-CHO, 5 »16 (1H, d, J = 5 cps, Cg-H), 5.64 (1H, d, J = 5 cps, C7-H) , 7.52 (broad S, phenyl).

30 '

Example 6 514 mg of disodium alpha (sulfophenylacetamido) cephalosporanate and 291 mg of KSCN are dissolved in 2.0 ml of water. To this mixture is added an additional 792 mg of 2,5-dimercapto-35 o

-3-phenyl-l, 3,4-thiadiazole. The mixture is stirred at 60 ° C

12 144177

ISLAND

for 8 hours. 10 ml of water is added to the reaction mixture.

The resulting precipitate is filtered to obtain 300 mg of crude crystals. The substance dissolved in the filtrate is, for the most part, 2,5-dimercapto-1,3,4-thiadiazole, which is used as starting material. The crude crystals are purified by column chromatography using "Amberlite XAD-2" resin to give 200 mg of sodium N- (7-ct-sulfo-phenylacetamido) -ceph-3-em-3- (2, -thio-3) , -phenyl-l ,, 3 ', 4' - thiadiazoline-5'-thio) -methyl-4-carboxylate.

7Dr 10 IRVmax (cm 1758 (β-lactam), 1676 (-CONH-), 1600 (-COONa), 1493, 1042 (-SO3Na)).

S H

NMR δ (D 2 O): 3.15 (2H, XH), 4.0 (2H,

H H

15 CH2-S-), 5.15 (2H, y-CHi S), 5.7 (1H, θ '-N-7.27 (5H, Ag), 7.5 (5H, -CHC).

Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 0.5

Bacillus subtilis 5

Sarcina lutea 2 Example 7 514 mg of disodium a-sulfophenylacetamido-cephalo-sporanate, 300 mg of KSCN and 500 mg of 2-mercapto-3-methyl-5-thio-1,3,4-thiadiazoline are dissolved in 2 ml of water . The mixture is adjusted to a pH of 5-6 and the reaction 30 is carried out at 60 ° C for 6 hours. The reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin to give 300 mg (50%) disodium N- (7-a-sulfophenylacetamido) -ceph-3-em-3- (21-thio) -31-methyl-1,3,3,4-thiadiazoline-51-thio) -methyl-4-carboxylate.

35

ISLAND

13 (1717 (IR-Vfftcm1): 1760 (β-lactam), 1677 (-CONH-),.

ΙΠ3.Χ 1604 (C02 ~), 1350, 1116, 1037 (-SC> 3Na)

S H

NMR (D2 O): 3.4 (2H, Xr), 3.70 (3H, S, 5 N-CH 3), 4.13 (2H, -> - CH 2 -S), 5.00 (1H, d, J = 4.6 cps,

H

- + - S), 5.20 (1H, S ir -CH 2), 5.75 (1H, d, J = 4.6 cps, _H_f_i), 7.4 - 7.6 (5H, phenyl) // ^ 10 o

Example 8 514 mg of disodium oc- (sulf ophenylacetamido) - cepha losporanate and 300 mg of KSCN are dissolved in 1 ml of water. Another solution prepared by dissolving 500 mg of 1- ^ phenyl-5-mercapto-tetrazole- in 1 ml of DMF is mixed with this solution. The mixture is adjusted to a pH of approx. 7th

The reaction is then carried out at 60 ° C for 6 hours with stirring. The reaction mixture is purified by column chromatography using "Affiberlite XAD-2" resin to give 150 mg (23%) of sodium N- (7-a-sulfophenylacetamido) -ceph-3-em-3- (1'- 51-phenyl-tetrazolyl-thio) -methyl-4-carboxylate.

IR ν 'KBr (cm -1): 1756 ((3-lactam), 1680 (-CONH-), max 1604 (COONa), 1496, 1357, 1040 (-SO, Na)).

25 J

S \ / H X

NMR S (D₂O): 3.2 (2H, Y), 4.15 (2H, CE CE₂ ~), 5.1 (2Ε-CEC, 4—S), 5.65 (1H, ), 7.35 (5H, NO / 30 + -CHIl), 7.52 (5H, N

Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 2.

Sarcina lutea 2 35 144177

ISLAND

14

Example 9 514 mg of disodium α- (sulfophenylacetamido) cephalo-sporanate and 291 mg of KSCN are dissolved in 1.0 ml of water. On the other hand, 6Q6 mg of 2-methylmercapto-5'-mercapto-5 -1,3,4-thiadiazole is dissolved in 0.8 ml of DMF. Both solutions are mixed and the mixture thus prepared is allowed to stand at 60 ° C for 6 hours. . The reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin to give 260 mg (42%) of sodium N- (7-a-10-sulfophenylacetamido) -ceph-3-em-3- (2 ') -methylmercapto-1 ', 3', 4'-thiadiazole-5'-thio) -methyl-4-carboxylate.

IR K K®r (cm -1): 1755 (β-laetam), 1675 (-CQNH-),

IttaX

1600 (-C00-), 1035 (-SO3Na) N

15 NMR 6 (DpO); 2.74 (35, S, 1), 3.37 - / NSCH3 3.54 (25, 3.8 - 4.4 (2H, 3 CH 2 -S-), 5.1 (25, HSH 4) W-mC., 4- S), 5-7 (15,), 7.5 (5Η, / -) -N cr 20 Minimum inhibitory concentration (γ / ml);

Staphylococcus apreus 2

Bacillus subtilis 5

Proteus vulgaris Eb51 5 Example 10 514 mg of disodium N- (7-a-sulfophenylacetamido) - cephalosporanate is dissolved in 2.5 ml of DMF. To this solution are added 630 mg of N-amino-2-mercaptopyridinium and 291 mg of KSCN. 2 ml of water is further added to homogenize the solution. The solution is allowed to stand at 6 ° C for 8 hours. The reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin.

The fractions containing sodium N- (7- [x-sulfophenyl-acetamido) -ceph-3-em-3- (N-aminopyridinium-2 L -thio) -35-methyl-4-carboxylate are lyophilized.

144177

ISLAND

IRV KBr (cm -1): 1760 O-lactam), 1673 (-CONH-), max 1610 (-CO2), 1575, 1038 (-SO3).

NMR ξ (D₂O): 3.37 (2H, C ^-H), 3.75 - 4.3 (2H,> -CH 2 -), 5.00 (1H, C , S, -CHU), 5.63 (1H, 5

H

C7-H), 7.5 (8H, phenyl and) 8.4 (1H, λ

-SAt © -s' ^ H

Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 5

Example 11 514 mg of disodium N- (7-α-sulfophenylacetamido) cephalo-sporanate, 350 mg of 1-phenyl-N-amino-5-mercaptotetrazolium and 200 mg of KSCN are dissolved in 1 ml of water. The solution is allowed to stand at 50 ° C for 10 hours. The reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin followed by lyophilization to give 320 mg of sodium N- (7-a-sulfophenylacetamido) - ceph-3-em-3- [51 - (1 '-phenyl-N-aminotetrazolium) thio) - methyl-4-carboxylate.

IR V> K 1 (cm-1): 1762 (β-lactam), 1677 (-CONH-), ΪΏα, Χ 1600 (-CO2)), 1498, 1395, 1222, 1039 (-SO ^). .

NMR ((D₂O): 3.3 (2H, Cj-H), 4.2 (2H ^ -CH₂-), 4.9 (1H, Cg-H), 5.0 5.6 (1H, C?-H), 7.63 (10H, 2 x phenyl)

Minimum inhibitory concentration (γ / ml): Escherichia coli 2 144177

ISLAND

16

Example 12 514 mg of disodium N- (7-a-sulfophenylacetamido) - cephalosporanate, 609 mg of 4-amino-2-mercapto-1,3,4-thiadiazolin-5-thione and 291 mg of KSCN are dissolved in 1 ml water. The mixture 5 is allowed to stand at 50 ° G for 23 hours. The reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin. The resulting fractions are lyophilized to give sodium N- (7-α-sulfophenylacetamido) -ceph-3-em-3- [5'- (2 '-thio-31-amino-1'; 3 ', 4' - 10-thiadiazoline) -thio] -methyl-4-carboxylate.

IR v'KBr (cm 1758 (β-lactam), 1675 (-CONH-),

IHEX

1600 (-CO2 "), 1353, 1218, 1039 (-SO3").

NMR S (D 2 O): 3.5 (2H, C 2 -H), 4.0 and 4.35 (2H, 1 -CH 2 -), 5.15 (2H, -CH <1 and Cg-H), 5 , 70 (1H, C?-H), 7.5 (5H, phenyl).

Minimum inhibitory concentration (γ / ml):

Escherichia coli 5 Example 13 514 mg of disodium α- (sulfophenylacetamido) cephalo-sporanate, 759 mg of 2-mercaptopyridine methiodide and 485 mg of KSCN are dissolved in 1.5 ml of water. This solution is adjusted to a pH of 6-7 by the addition of solid NaHCO 3.

The mixture is allowed to stand in a thermostat at 60 ° C

for 12 hours. The reaction mixture is purified directly by column chromatography using "Amberlite XAD-2" resin (eluent: water) to give sodium N - (7-a-sulfophenylacetamido) -ceph-3-em-3- (N-methyl) 30-pyridinium-2'-thio) methyl-4-carboxylate.

U4177 17

ISLAND

VTDv · —1 IR V! ” (can): 1760 (β-lactam), 1674 (-CONH>

• IIlciX

C = N), 1610 (-CO 2 ", C = C), 1037 (-SO 3 Na).

NMR <f (D20): 2.83 (3H, S, N + -CH3), 5.05 (1H, S, fiP-CHO, 7.5 (phenyl), 7.5 - 8.5 ( Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 2

Bacillus subtilis 5

Pseudomonas aeruginosa 2 Example 14 514 mg. disodium α-sulfophenylacetamido-cephalo-sporanate, 757 mg of 4-mercaptopyridine methiodide and 485 mg of KSCN are dissolved in 1.5 ml of water. The solution is adjusted to a pH of 6-7 by the addition of solid NaHCO 3. The mixture 20 is then allowed to stand in a thermostat at 60 ° C for 12 hours. After completion of the reaction, the reaction mixture is purified by column chromatography using "Amberlite XAD-2" resin (eluent: 10% aqueous ethanolic solution) to give 200 mg of crystals of sodium N- (7-a-2 $ sulfophenylacetamido ) -ceph-3-em-3- (N-methyl-4'-pyridinyl) -21-thio) methyl-4-carboxylate.

VT3r IR (can): 1765 (β-lactam), 1673 (CONH-,

_i_ lucLX

C = N) 1629 (C00-, C =), 1500 (C = C), 1102, 1034 (-SO3Na).

NMR ((DgO): 2.65 (3H, S, N + -CH3), 3.25 (2H,>> ^ H), 5.08 (1H, S, tP- CH <J, 5.2 ( 3H, - + - + - -CH 2 -S-), -H ___ 5.68 (1H,), 7.45 (5H, phenyl), 7.7 (2H, broad

H J

35 -S- ^ N + -CH3), 8'4 (2h 't> red - {9 ^ + - CH3).

Η H

U4177 18

ISLAND

Minimum inhibitory concentration (γ / ml):

Staphylococcus aureus 1

Pseudomonas aeruginosa 5 Example 15 517 mg of disodium 7- (α-sulfophenylacetamido) cephalosporanate, 522 mg of 2-mercapto-2-thiazoline methiodide and 1.75 g of KSCN are dissolved in 2 ml of water. The solution is adjusted to a pH of 6.5-7 and heated to 60 ° C for 7 hours. The reaction mixture is purified by column chromatography using "Amber li te XAD-211 resin to give sodium 7- (a-sulfophenylacetamido) -3 - (3'-methyl-2'-thiazolium-2'-thio) -methylceph-3-em-4 - carboxylate.

IR V (cm -1): 1760 (& lactam), 1665 (-CONH-),

TtlclX

1610 (002 "), 1530, 1390, 1355, 1320, 1210, 1040 (-SO3"), 700 NMR S (D, 0): 2.90 (3H, S, N-CH3), 3 - 4, 2 (6H, Æ

m, s yH

2 X 5 V, H, X), 5.10 (1H, S O-CHO, 5.17 (1H,

H 'nH

d, J = 4.2 cps, Cg-H), 5.74 (1H, d, J = 4.20 cps, Cy-H), 7.52 (5H, broad, phenyl).

Minimum inhibitory concentration (γ / ml): 25

Staphylococcus aureus 2

Bacillus subtilis 5

Sarcina lutea 5

Pseudomonas aeruginosa 5 30

Example 16 (1) 1.84 g (0.0069 mol) of 7-aminocephalosporanoic acid is suspended in 25 ml of water with stirring while cooling on ice. To this suspension is added dropwise 6.8 ml of 1 N 35 aqueous NaOH solution to complete solution herein.

Then 10 ml of ethyl acetate solution of α-sulfo-3-thiophene-144177 is added

ISLAND

19 acetyl chloride is added dropwise to the mixture, this solution being prepared by the following method: 1.5 g (0.0068 mol) of alpha-sulfo-3-thiophenacetic acid is suspended in 10 ml of ether, 5.5 ml of thionyl chloride is added dropwise thereto at room temperature with three drops of dimethylformamide and the resulting solution is stirred for 5 hours. After the mixture is stirred for 30 minutes, the aqueous layer is recovered, adjusted to a pH of 6.5 by saturated aqueous NaHCO 3 solution and lyophilized to give 3.6 g of crude crystals. These crude crystals are purified by "Amberlite XAD-2" column chromatography to give 1.6 g (42%) of disodium 7- (α-sulfo-3'-thiophenacetamido) cephalosporanate crystals.

IR1 max (cltl_1) 1750 (3-lactam, -COCH), 1677 (-CONH-), T605 (-COONa), 1225 (-SO3Na, -OAc), 1043 (-SO3Na).

NMR § (D20): 2.13 (3H, S, -OCOCH3), 3.54 (2H,

Sv Η H

b, X), 5.10 (1H, d, J = 4.7 cps, H-S), 5.24 (1H, S,

20 H N

-CH₂), 5.70 (1H, d, J = 4.7 cps), 7.38-0.67 (3H, m, O-25 -3 (2) 600 mg (4 x 10 mole) 2,5-dimercapto-1,3,4-3-thiadiazole, 260 mg (0.5 x 10 mole) disodium 7- (α-sulfo-3-thienylacetamido) cephalosporanate and 194 mg -3 (2 x 10 mol) KSCN is dissolved in 1.5 ml of water. The solution is left at 60 ° C for 8 hours. The reaction mixture is purified by column chromatography with "Amberlite XAD-21" resin to give sodium N- (7- α-sulfo-3-thienylacetamido) -ceph-3-em-3- (5'-mercapto-1 ', 3', 41-thiadiazolyl-2-thio) methyl-4-carboxylate.

ISLAND

144177 IR V max (citl_1): 1755 (3-lactam), 1660 (-CONH-), 1605 (-COONa), 1040 (-SO2Na).

S Η H

NMR δ (DO): 3.5 (2H, X), 5.2 (1H, -j-S),

'5 N

5.2 (1H, S, -CH <3 SO) Na, 5.65 (1H - **), 7.4 - 7.6 (3H, thiophene). ^ 0

Example 17 0.406 sodium 7-aminoceph-3-em-3- [5- (2'-thio-11,3 ', 4'-thiadiazolyl) -thio] methyl 4-carboxylate and 0.17 dissolve sodium bicarbonate in 7 ml of water. To the resulting solution, 3 ml of a chloroform solution containing 0.234 g of a-sulfophenylacetyl chloride is added dropwise with cooling.

After completion of the drop, stirring is continued under cooling for 40 minutes to complete the reaction. After removal of the organic layer, the aqueous layer is adjusted to a pH of 6 and purified chromatographically using a resin column "Amberlite XAD-2" ® 20 (trade name of Rohm & Haas Co., USA, 16 x 700 mm, water as eluent, control with a UV spectrometer (240 µl)). The fractions containing the desired cephalosporin are collected and then lyophilized to give 500 mg of 7- (α-sulfophenylacetamido) -3- [5 (2'-thio-25-1,3,3-41-thiadiazole) -thio] - methyl-3-cephem-4-carboxylate.

IR ^ (cm -1): 1750 (β-lactam, C = O), 1670 (-CONH-), 1600 (-C00-), 1530 (-NC = S), 1285 (CSNH), 1400, 1360 , 1215 (SC> 2), 1045 (-S03H) NMR. (6OMc, D20): 3-4 (2H, C2-H2), 5.1 (1H, S, side-chain -CH-), 5.06 (1H, doublet, J = 5 cps,

Cg-H), 5.62 (1H, doublet, J = 5 cps, C7 ~ H), 7.5 (5H, broad S, phenyl -H).

35 21

ISLAND

HA 177 In the above analyzes in the examples, S means singlet, d doublet, m multiplet, 5 b wide.

A representative selection of the compounds prepared by the process of this invention has been compared with respect to the antimicrobial action with compounds known from the specification of Danish Patent Application No. 606/71. The results are summarized in the following table, which shows that the compounds of the present invention are clearly superior to the known compounds.

Table 15 - - R'-CHCONH __ ^ \ kNa J — v ^ L-ch2r COONa 20 1.1 '' 1-111 '! 1 '1 II' 1 '· "ί · H' * I i 1 '<I I III" 1) MIK (pg / ml) vs. Ps. aeruginosa NCTC 10490

R 'R MIK

25 rivers. no.

606/71 H -OCOCH3 ^ 50 1 "·“ · "1 i 111 * l" I. II I I ^ .IJ I - ... I '. "»! 'II p 1 C4Hg- -OCOCH3 50 O- -C? 0

According to N-N 35 invention- -S- [l JLsNa 20 '-' s -S-l- <"10 ^ \ = /

ISLAND

22 144177 ao i-ο-, ι ~ o -p CH3 10 ^) - -S- <f ^ -CH3 5 CH-, O -> 4h 15 _____ i / —tv N - N-NH9 O -u .

20 N N

H -S- lLs Ji-SNa 10 '. ....... IL si— -s-lsi-5 · 13 20 2b

2) MIK µg / ml) against E. coli NIHJ

30:

R 'R MIK

Ans. No. -0C0CH-. 50 606/71 \ _ / ~

up

island

ISLAND

23 144177 I o- I ajP F ~ 5__________

Ι £? ^ Β / Γ ~ \ Ψ-N — CH

£ 2 = O YES,

10 N-N

-S-U N

O 1 o O -G-

O ~ Q

nh2 25 / γ-λ -s-ifi ~ NH2 α 6 1 '30

Claims (3)

Analogous Process for Preparation of 7- (α-Sulfoacetamido) -3- (Heterocyclyl-Thiomethyl) -Ceph-3-Em-4-Carboxylic Acid Derivatives of General Formula 5 R1-CHCONH -. CH2SR2 (D COOH wherein R · is a hydrogen atom, phenyl or thienyl, 2 and R is a pyridyl, pyrimidyl, imidazolyl, thiazolyl, thiadiazolyl or tetrazolyl group which may be substituted by methyl, amino, thio, mercapto , methylthio, phenyl and / or acetamido, or pharmaceutically acceptable salts thereof, characterized in that (a) a cephalosporin of the general formula 20 R 1 -CHCONH-r__v // X SO, H J-Nv LcH90CCH, (II) / ooi COOH 25 wherein Rx has the above meaning or a salt thereof is reacted with a heterocyclic compound of the formula R 2 -SH (III) 2 wherein R is as defined above, or a salt thereof, or (b) an amino-cephalosporanoic acid derivative of the general formula