CH629500A5 - Processes for preparing novel cephalosporins. - Google Patents

Description

The present invention relates to a process for the preparation of new synthetic cephalosporins which are useful as antibacterial agents by both oral and parenteral administration in the treatment of infectious conditions in humans and animals caused by gram-positive and gram-negative bacteria. These compounds can also be used as feed additives in cattle feed and as agents for the treatment of mastitis in cattle.

These products mentioned are condensation products of aldehydes with 3-thiolated cephalosporins which have a 7-hour a-amino-phenylacetamido substituent, which substituent in the benzene ring is substituted with one or two particular groups which groups do not react with aldehydes wise but with a para-hydroxy group to react. The term “3-thiolated cephalosporin” is understood to mean a derivative of cephalosporanic acid in which the acetoxy group has been displaced by a thiol, the 3-acetoxmethyl group having been converted; in these thiols the mercapto group is 1,2,3-triazol-5-yl, tetrazol-5-yl, l, 2,4-thiadiazoi-5-yl, 1,3,4-thiadia-zol-2-yl , l, 3,4-oxadiazol-2-yl or 1,2,4-triazol-5-yl substituted, the ring being optionally substituted with one or two lower alkyl groups.

The condensation products resulting from the process in the form of their sodium or potassium salts have a desired solubility, stability and absorption. The preferred types of compounds hydrolyze rapidly and completely in the body, regenerating the original amphoteric 3-thiolated cephalosporin; this is not the case with corresponding condensation products with acetaldehyde, formaldehyde or acetone, which hydrolyzes completely but at an undesirably low rate.

By the process according to the invention, the free acid or the sodium or potassium salt of a condensation product of a) an aldehyde of the formula R2-CHO, in which R2 is carboxyl or 2-furyl or an aliphatic, aromatic or heterocyclic radical, on which a strong Acid group is bound in the form of a free acid or as a sodium or potassium salt, with b) an amphoteric 3-thiolated cephalosporin which has a 7-position, in the a-position by 4-hydroxyphenyl. which optionally contains hydroxy, methyl or methoxy in the 3-position, contains substituted a-aminoacetamido group and whose zwitterion form has an aqueous solubility of less than 125 mg / ml. In one embodiment of this process, an aldehyde-5-formyl-2-furansulfonic acid, o-benzaldehyde sulfonic acid, 4-methoxybenzaldehvd-3-sulfonic acid, 4-hydroxybenzaldehyde-3-sulfonic acid, o- and p- Formylphenoxyacetic acid, 5-formyl salicylic acid, p-formyl cinnamic acid, glyoxalic acid, phthalaldehyde carboxylic acid or p-formyibenzoic acid, sodium salt of acetaldehyde sulfonic acid or of acctaldehyde disulfonic acid, are used.

Derivatives of various a-amino-cephalosporins with nitro-substituted heterocyclic aldehydes are described in US Pat. No. 3,647,781. Reaction products of various a-amino-cephalosporins with formaldehyde are disclosed in South African patent 72/8475, with acetaldehyde in South African patent 72/8474 and with various aldehydes and ketones in South African patent 72/8476.

Derivatives of cephalosporins with a 7-position a-amino group in the acylamido group which have been reacted with an aldehyde, but which have been limited to methyl or acetoxy-methyl 3-position, are disclosed in U.S. Patent 5,880 842 and 3 887 546 as well as in the Farmdoc 49804W.

The reaction products of acetone with various a-amino-cephalosporins are described in the following patent literature:

io (1) with cephaloglycine in U.S. Patent 13,303,193;

(2) with cephalexin in U.S. Patent 3,714,146 and UK Patent 1,314,758 and U.S. Patent 3,780,028;

(3) with 7- [a-amino- (2'-thienyl) acetamido] cephalosporanic acid in U.S. Patent 3,311,621;

15 (4) with certain ring-substituted cephaloglycins, in U.S. Patent 3,464,985, and

(5) with certain ring-substituted cephalexins and cephaloglycins, in U.S. Patents 3,489,750,3,489,751 and 3,489,752.

2o In a more distant relationship are the intermediate substances that are produced if a cephalosporin core, e.g. B. 7-ACA or 7-ADCA, is aeylated with a reactive derivative of an a-amino acid, in which a-amino acid the a-amino group previously with a ß-diketo compound, such as methyl acetoacetate, 25 methyl acetoacetamide or acetylacetone was protected. These intermediate substances are listed, for example, in Farmdoc 22850W and 60669V.

In the field of penicillins, a cc-amino group in penicillins containing 7 acylamido substituents, e.g. B. Ampi-30 cillin, with ketones and aldehydes apparently were first developed by Johnson and Mitarb. (U.S. Patent 3,198,804) and Granatek (U.S. Patent 3,198,788). Similar reaction products made from various such penicillins by reacting the same or different aldehydes and ketones 35 were later reported in U.S. Patents 3,230,214,3,316,247 (Diketone), 3,325,479 (Diketone), 3,489,746.3,549 746, 3 558 602.3 635 953.3 641 000.3 647 781 (which include some cephalosporins), 3 725 389.3 780 028.3 784 562 (diketones), 3 886 140.3 888 848.3 905 966 and 3 904 604 and io UK 1 267 936.

The present process according to the invention thus relates to the preparation of the compound of the formula I.

"O-i

50

55

-sr wherein A is hydrogen, hydroxyl, methyl or methoxy, R1 is hydrogen, sodium or potassium, R2 carboxyl or 2-furyl or an aliphatic, aromatic or heterocyclic radical on which a strongly acidic group, in the form of the 60 free acid or is present as the sodium or potassium salt, and R-11.2,3-triazol-5-yl, tetrazol-5-yl, 1,2,4-thiadiazol-5-yl, l, 3,4-thiadiazol- 2-yl. 1, 3,4-oxadiazol-2-yl or 1,2,4-triazol-5-yl, each of which is optionally substituted with one or two lower alkyl groups, preferably 1-4 65 carbon atoms. In a preferred embodiment, compounds are obtained in which the carbon atom on the benzene ring, para to the hydroxyl group, has the D configuration.

629 500

S

The present process according to the invention is based on amphoteric cephalosporins of the formula II water-soluble, pharmaceutically acceptable derivatives of ch2-s-e3

II

where A, R1 and R3 have the above meaning. In a preferred embodiment, the carbon bearing the a-amino group has the D configuration. These are derivatives which (1) give real solutions for parenteral administration after the addition of water, (2) have acceptable thermal stability in the solid state, (3) show a useful lifespan of at least several hours at room temperature in aqueous solution and (4) cause little or no muscle or vein irritation when injected intravenously or intramuscularly. The present invention particularly leads to sodium and potassium salts of the reaction products called amphoteric cephalosporins with aldehydes and preferably with furfural dvd or an aldehyde which contains an acid group, e.g. B. 2-furansulfonic acid group contains.

The compounds produced according to the invention have a desired solubility, stability and absorption. The preferred species hydrolyze rapidly and completely in the body, regenerating the original amphoteric cephalosporin of Formula II. This is not the case with corresponding derivatives, made from formaldehyde, acetaldehyde or acetone, which hydrolyze completely but at an undesirably slow rate. The compounds produced according to the invention thus overcome the disadvantages and the problems which are posed by the instability towards a high pH and by an often occurring relative insolubility in their zwitterion form of the amphoteric cephalosporins of the formula II.

In particular embodiments of the present method according to the invention, the aldehyde with an acid function is selected from the following group of compounds:

CHO

0

H

H-C-

ß

"LT

S03 Well

5-Formvl-2-furansulfonic acid sodium salt.

CHO-

SN SCUH

15

25th

30th

35

40

45

50

60

65

o3h

OCH

4-methoxvbenzaldehyde-3-sulfonic acid,

HO

s ° 3h

4-hydroxybenzaldehyde-3-sulfonic acid,

CHO CHO

OCHoCOOH

o- and p-formylphenoxyacetic acid,

coca

GH

I.

0CH2C00H

CHO

5-formyl salicylic acid,

o-Benzaldehvd-sulfonic acid.

CH = CHC00H

p-formyl cinnamic acid.

629 500

OCH - COOH

Glyoxalic acid,

CHO

COOH

Phthalaldehyde acid,

CHO

, 0

or or

COOH

p-formylbenzoic acid,

OHC - CH, S03Na acetaldehyde sulfonic acid sodium salt and OHC - CH (S03Na) 2 acetaldehyde disulfonic acid sodium salt.

By means of further preferred embodiments of the method according to the invention, compounds of the formula I in which A is hydrogen, R2 is selected from the aldehydes mentioned above and preferably

N_S0, well

J is and R 'for ri

SI N stands for

H

The compounds produced according to the invention thus solve the problem of formulating relatively water-insoluble, ie. H. less than 125 mg / ml amphoteric 3-thiolated cephalosporins for intravenous injections, where real solutions with a concentration of about 250 mg in only 1 or 2 ml for bolus injections are required. Furthermore, such a dosage form can be made by dry filling, i.e. Vials containing only powder are provided, which is prepared, especially for use by adding sterile water, but, in order to be usable in such a case, it only has to be completely soluble in a few minutes in that period.

In addition, if a larger volume of infusion liquid is added during the preparation for intravenous drip enema up to a concentration in the range of 10-25 mg / ml, the compound must not exceed 10 Cc of its bioactivity within the 4-6 hours required for the infusion to lose.

Amphoteric 3-thiolated cephalosporins, which in their zwitterion form have an aqueous solubility of less than about 125 mg / ml, are of course not suitable for this purpose and, moreover, in contrast to their non-amphoteric counterpart, such as cephalothin, etc., they can be used are not converted into customary, soluble sodium salts after the required pi I is so high that it leads to decomposition, and moreover the amino groups free from this pi I are such that they catalyze decomposition.

The above-mentioned process according to the invention for the preparation of a compound of the formula I defined above is characterized in that (1) a suspension in water or an inert organic solvent of an amphoteric cephalosporin of the formula 11 or a solvate thereof, with an aldehyde or Formula R2-CHO, wherein R: has the above meaning, and treated with a sufficiently soluble sodium or potassium base or triethylamine in water or an inert organic solvent to adjust the pH of the reaction mixture between 5.5 to 8 and preferably 6, Bring 2-7.2 into solution to form the salt of formula I, and thereafter (2) 15 from this solution of the salt or the acid of formula I recover.

The compounds of the compound I mentioned have an asymmetric center on the carbon linked to two nitrogen atoms. Such compounds of the formula I can be present in form 20 of the DL mixture or their individual D or L isomers.

The cephalosporin used in step (1) as a starting material in the above process can be any form of the amphoteric cephalosporin of formula II, including the free zwitterionic acid or a zwitterion called hydrate or solvate.

The concentration of amphoteric cephalosporin starting material is not critical and good results have been obtained with concentrations between about 25-300 mg of cephalo-30 sporin starting material per ml of solvent. The starting material is preferably ground and sieved to a finely divided state, but most preferably ground to a particle size of less than 200 mesh to increase the surface area and hence the reaction rate.

The amphoteric cephalosporin starting material is slurried in water or an inert organic solvent. The organic solvent is a solvent for the alkali metal salt end product, soluble with the aldehyde, chemically inert to the cephalosporin starting material and end product and, moreover, easily removable from the end product, such as by mild drying. Examples of such organic solvents that can be used are dimethyl sulfoxide and dimethylformamide. Because of the difficulty in removing the residual organic solvent 45 from the final alkali metal salt product, the starting material is advantageously supplied as an aqueous suspension.

After the cephalosporin starting material is obtained in suspension, the desired alkali metal salt of formula I is dissolved in solution by adding the aldehyde and an amount 50 of water-soluble sodium or potassium base in an amount sufficient to bring the pH of the reaction mixture to between 5.5- 8 to bring up. As soon as the pH was adjusted within this range, the alkali metal salt of the reaction product forms from the amphoteric cephalosporin and the aldehyde and goes into solution.

The temperature at which step (1) is carried out is not critical. The reaction can be carried out at room temperature; however, higher or lower temperatures can be used and preferred are temperatures in the range of 50-60 ° C.

About 1 mole of the aldehyde is required per mole of cephalosporin starting material, but the aldehyde is advantageously somewhat in excess of the necessary theoretical amount, i.e. H. in a slight molar excess, 65 is added to ensure a complete reaction. The most preferred ratio of aldehyde to cephalosporin starting material is about 1.3-1.4: 1 and often the 1: 1 ratio is preferred.

35

40

629 500

10th

The alkali metal base is one that is capable of

Form sodium or potassium ions and adjust the pH of the reaction mixture to between 5.5 and 8, most preferably between about 6.2 to 7.2. Preferred bases are sodium or potassium hydroxide because of their desired solubility properties. The R'-S portion of Compound 1 can be cleaved off at a higher pH. For this reason, the base is added to the reaction mixture in such a way that the pH does not exceed 8. Preferably the base is used in the form of an aqueous solution and is slowly added to the reaction mixture with stirring until the reaction is complete, which can be determined by pH measurement and by formation of a solution or a fast solution. The amount of base used is not critical, but preferably 1 mole base per mole of the cephalosporin starting material is used.

For best results, the solution obtained after completing step (1) is filtered to filter solid contaminants before extraction (2). Before filtering, the solution may optionally be carbon treated with activated carbon to aid in the removal of any colored contaminant.

The desired product of formula I is then obtained from aqueous or non-aqueous solution, preferably either by precipitation or by lyophilization. The alkali metal salt can be precipitated by adding an organic solvent in which the desired salt is insoluble. d. H. of a counter solvent. Examples of such counter solvents are isopropanol, n-propanol, t-butanol and acetonitrile.

The solvent to be used in the precipitation is usually said to be one which can be conveniently removed from the final product under conditions which do not result in significant decomposition of the alkali metal salt. The most preferred anti-solvent is isopropanol. The anti-solvent can be added to the solution resulting from step (1) or, according to an alternative, and preferably the solution containing the desired alkali metal salt is added with stirring to a large excess of the anti-solvent. The formula alkali metal salt can then be collected by filtration, with a suitable organic solvent, e.g. B. isopropanol. washed and dried in the usual way, e.g. B. in vacuum at 50-56 ° C for 24-48 hours or in air at 60 ° C for 48 hours. According to an alternative method for obtaining the end product by precipitation, the salt of the formula I can also be obtained by lyophilizing the solution prepared in stage (1). _

An alternative embodiment of the process for the preparation of the compounds of Formula I involves (1) forming a suspension of the amphoteric cephalosporin or a solvate or hydrate thereof in a suitable inert organic solvent, said solvent being a solvent for the triethylamine salt of the aldehyde reaction product of the amphoteric Cephalosporins and a nonsolvent for the alkali metal salt of Formula I is: (2) treating the suspension with the aldehyde and enough triethylamine to form in solution the triethylamine salt of the alde hyd reaction product of the amphoteric cephalosporin; (3) Precipitation of the desired alkali metal salt of formula I from the solution by adding a solvent-soluble sodium or potassium base.

The starting material is conveniently in an inert organic solvent which is a solvent for the triethylamine salt or the aldehyde reaction product of the amphoteric cephalosporin, but which is a non-solvent for the desired alkali metal salt of formula I. suspended. The selected solvent for stage (1)

10th

should advantageously be easily removable from the end product under conditions. which do not lead to any significant decomposition of the end product. Suitable mare solvents (1) can be determined using a simple test sample.

The suspension formed in step (I) is then conveniently treated with an aldehyde, preferably with a molar excess and most advantageously with about 1.3-1.4 moles of the aldehyde per mole of the cephalosporin starting material, and with sufficient triethylamine. treated for formation in solution of the triethylamine salt of the cyclic reaction product of the aldehyde and the amphoteric cephalosporin. The reaction mixture is advantageously stirred for at least about 30 minutes to effect a complete reaction. The amount of triethylamine used is not critical, but advantageously about 1 mole per mole of cephalosporin starting material is used. The reaction of stage (2) is best carried out at room temperature, although higher or lower temperatures than this 20 can be chosen in anticipation of an increased or weakened reaction time.

After formation of a solution or a fast solution in step (2), the reaction mixture is advantageously carbon-treated and filtered, as was stated in the first-mentioned process according to the invention.

The desired alkali metal salt of formula I can then be obtained from the solution of step (2) by adding a solvent-soluble potassium or sodium salt. The preferred salts are potassium or sodium salts of organic acids with between about 2 and 18 carbon atoms, e.g. B. Solvent-soluble salts of acids such as 2-ethylcaproic, capronic, oleic, glycolic, propionic, acetic acid, etc. Preferred salts for the methanol solvent system are sodium or potassium 2-ethylhexanoate and the most preferred solutions thereof Salts in a metal-miscible organic solvent such as isopropanol. The most preferred alkali metal salts are solutions of sodium or potassium 2-ethylhexanoate in isopropanol. The alkali metal salt is added, preferably slowly and with stirring, in an amount sufficient to obtain the maximum amount of precipitate from the solution. After complete precipitation, the reaction mixture is stirred, advantageously for at least about 1 h and then filtered. The precipitate is washed with a suitable organic solvent, e.g. B. methanol, washed and dried in a conventional manner, e.g. B. by vacuum drying at 50-56 ° C for 24-48 hours or by air drying at 60 ° C for 48 hours.

However, the second method can also be carried out in stage 2 without using the triethylamine. In this modified process, the suspension of the amphoteric cephalosporin or a solvate or hydrate thereof, preferably the methanol or propylene glycol solvate or hydrated forms, and most preferably the methanol solvate. in an inert organic solvent which is a nonsolvent for the product of Formula I. preferably methanol, and the suspension is then treated with aldehyde, preferably in a molar excess, and with a solvent-soluble sodium or potassium base, said base being added in an amount sufficient to bring the pH of the reaction mixture to between about 5.5 and 8 set. It is preferred to use the above-mentioned sodium or potassium salts as bases, since these are also advantageous in the second process. In this modified process, the cephalosporin starting material goes into solution and the insoluble alkali metal salt mostly precipitates instantaneously. Since a solution is not obtained after the reaction has ended, the reaction mixture is advantageously stirred and brought to about 45-50UC over a period of time.

25th

30th

35

• 40

45

50

55

60

65

11 629 500

range from up to about several hours for maximum acidity, solubility, etc. Generally it was found

to secure male yield of final product. The test obtained obtained that the desired pH was about 5 to 6 after contact with

The product is then filtered, washed and dried and water then occurs if the solid organic acid in one obtains the desired salt of formula 1. Amount of about 4-6% by weight of the compound of formula I

The alkali metal salts produced according to the invention can be used for 5.

pharmaceutical formulations of the amphoteric cephalosporin are processed, which are characterized by an acceptable The compounds of formula I as well as those with these thermal stability in the solid state, a high solubility in the above-mentioned compositions are strong water or a satisfactory aqueous stability, antibacterial agents, which cause oral as well as small or no muscle or vein irritation after intra- io parenteral administration in the treatment of infectious venous or untramuscular injection and suffering in humans and animals caused by numerous excellent in vivo and in vitro antibacterial effectiveness gram-positive and gram-negative bacteria. The compounds and compositions mentioned in relation to a variety of gram-positive and gram-negative are the same bacteria. if as feed supplements for animals and as a means for the

The sodium and potassium salts of formula I can be used in treatment of mastitis in cattle of high value.

Dissolve water, forming relatively concentrated salts and mixtures of

Solutions of at least 250 mg / ml effectiveness. Concentrated salts and organic acids can be formulated as pharmaceutical ions of 250 mg / ml effectiveness with pH 5.7-6.8 of these salts. Compositions which, in addition to the active, have an acceptable aqueous stability. Component a pharmaceutically acceptable carrier or

The diluents according to the invention hydrolyze in dilute aqueous solution. The compounds mentioned can be administered either orally or parenterally, but because of the compounds produced to the amphoteric parent

Cephalosporin of formula I. In acidic aqueous solutions of their higher solubility in water, they are particularly useful for any concentration, the compounds hydrolyze rapidly parenteral administration. In the treatment of cephalosporin called stem. This property makes bacterial infections in humans the link

these are useful for the purposes of purifying the amphoteric genes and compositions in an amount of about 5-20 mg /

Stock antibiotics, e.g. B. in solid form they can with non-kg per day in divided doses, for. B. three or four times aqueous solvents for the purpose of removing which are administered daily in such.

Soluble contaminants are washed. The starting compounds of formula II mentioned, counter

and after such treatment they can easily be converted into their D-form, too, by common and often amphoteric strain antibiotics. 30 produced by specific processes, which are described in the following

The activities of the compounds of the formula I are disclosed in the following:

substantially equivalent to that of the amphoteric parent cepha- US3 641 021, US3 899 394, US3 855 213, US3 867 380,

losporins. South Africa 73/4055, Belgium 776 222 (Farmdoc 38983T), Belgium

The compounds produced according to the invention can be 810 477 (Farmdoc 57268V), FRG 2 364 192 (Farmdoc 49048V),

pharmaceutical compositions processed, which are 35 BRD 2 500 386 (Farmdoc 49692W), Belgium 814 727 (Farmdoc after combination with water as parenterally administrable 82562V), BRD 2 404 592 (Farmdoc 57268V).

Antibiotic formulation are suitable, called composition. An alternative method for the preparation of the amphoteric cephalosporins used as starting material contains a compound of formula I and a solid pharmaceutical material consists of table-acceptable water-soluble organic acid, whereby the suitable p-hydroxy-2-phenylglycine, which said organic acid is present in such an amount, 40 may contain an additional substituent and wherein the a - that the pH of the formulation after contact with water between amino group during the acylation is suitably about 5.5 and 7.5. is protected for the side chain acid, e.g. B.2-phenylglycine to substitute the above mixture of salt of formula I and the or tetrazole-acetic acid, which method organic acid in the above composition can be previously mixed with water in U.S. Patent Nos. 3,813,388 and 3,759,904 to include a higher one Konzentra- «And3 850 916 was used to provide either 3-thiolated ion, i. H. up to at least 250 mg / ml for parenteral cephalosporins or 7-substituted cephalosporal administration of suitable solution. A composition rans acids, in which the 3-acetoxy group can then be prepared by that, so that one can displace a compound of the desired thiol, as for example formula I with a sufficient amount of a pharmaceutically in US Pat. No. 3,757 012 and 3 757 015 is included. In the following examples, the starting substances are mixed with, so that the pH of the composition is named after contact with code names such as BL-S640, BL-S643 and BL-S689. BL water is between about 5 and 6. S1640 refers to 7- [D-cx-amino-a- (p-hydroxyphenyl) -acet-

The organic amido] -3- (1,3,3-triazol-5-ylthiomethyl) -3-cephem-4-carbon used in this composition

Acid can be any non-toxic, water-soluble, solid organic acid and such acid is also called cefatrizine. BL-S643

see be acid. Examples of such suitable acids include 55 refers to 7- [D-a-amino-a- (p-hydroxyphenyl) acetamido] -

Citric acid, tartaric acid, glycine hydrochloride, ascorbin 3- (2-methyl-l, 3,4-thiadiazol-5-yl-thiomethyl) -3-cephem-4-car-

acid, succinic acid and the like. The most preferred bonic acid and also called cefaparole. BL-S689 is the 7- [D-a-

Acid for use in the above composition is amino- (3'-methoxy-4'-hydroxyphenyl) acetamido] -3- (1,2,3-

the citric acid. triazol-4-ylthiomethyl) -3-cephem-4-carboxylic acid.

The exact amount of the compound of formula I and the 60 The inventive method described above is in solid organic acid depends on the physical and the following examples as preferred embodiments closer chemical properties of the selected acid, e.g. B. set out by the.

629 500

12th

example 1

CKg-S

To 0.34 ml of furfural (2-furfuralde-hyd) dissolved in 10 ml of water was added 2 g (1 equivalent) of the methanol solvate of 7- [Da-amino-a- (p-hydroxyphenyl) acetamido] - with stirring 3- (1,2,3-triazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid added. Thereafter, 4N sodium hydroxide solution was poured in with vigorous stirring to maintain a pH of 6.2-6.5. The resulting solution is then held at room temperature for 1 h and then lyophilized to give the sodium salt of the product as a solid. Examination of the material showed that the IR-NMR spectra 2S correspond to the product, that the β-lactam and triazole ring are intact.

COgNa

Example 2

The procedure of Example 1 was repeated except that the starting methanol solvate was replaced with an equivalent amount of 7- [Da-amino-a- (p-hydroxyphe-nyl) acetamido] -3-cephem-4-carboxylic acid sesquihydrate. A sodium salt identical to that of Example 1 was produced.

Example 3 Preparation of BL-S1052 of Formula ch2s-

COONa

I.

H

.h2o this is the sodium salt of the reaction product of 2-furfural aldehyde and BL-S640. 45

Method:

1) 2.7 / 10 ml (3.13 g, 1.1 equivalents) of 2-furfuraldehyde are dissolved in 50 ml of rapidly stirred water at 20-30 ° C.

2) 15 g of the methanol addition product of BL-S640 are added within a 10-minute period with associated or simultaneous addition of 40% sodium hydroxide to a pH of 5.5-6.0, the latter a pH of over 6.5 must not be reached. A slightly orange-colored solution or a fast solution is obtained. 55

3) Filter this solution through suitable filters to remove particles. Fever-producing substances and bacteria. Levels 1 and 2 should be completed within 2 hours.

4) Lyophilize for 48 hours and then maintain the vacuum on the solids at 50-56 ° C for 24 hours. The 60 solid that results is BL-S1052; this can also be obtained from the level 3 solution by precipitation from 15-20 volumes of sterile isopropanol.

Properties of BL-S1052 65

1) Bio testing is = 775-800 mcg / mg.

2) IR-NMR '= a - well defined, solid, b) at about 60 mg / ml in DiO two products appear, namely a 40% cyclic adduct and a 60% non-cyclic adduct. c) ß-lactam and 3-triazole rings intact.

3) Solubility is greater than 400 mg / ml.

4) Paper strip chromatography shows a single zone at Rf of BL-S640 at a concentration of 0.2 mg / ml.

5) Liquid chromatography at concentrations of 1 mg /

ml.

Time in h

% free BL-S640 present

0

69.2

1

74.2

2nd

94.2

6) Analytical data

Found based on theory

Dry

substance

7c ILO KF

5.35

-

-

r-c C

47.45

50.2

49.2

c / c H

3.72

3.4

3.39

Cr N

14.45

15.29

14.95

7c p

10.34

10.92

.11.4

c / c ash as Na

2.18

2.51

4.1

13

629 500

Antibiotic spectrum in nutrient broth organism MIC (incg / ml)

BL-S640 BL-S 1052 '

S. pneumoniae5 '

(10 •) ''

A9585

0.06

0.06

St. pyogenes *

(10 ') **

A9604

0.03

0.03

S. aureus Smith

(10 •)

A9537

0.13

0.13

S. aureus + 50% serum

(10 ')

A9537

2nd

2nd

S. aureus BX1633

(10 ")

A9606

0.25

0.25

S. aureus BX1633

(10;)

A9606

4th

2nd

S. aureus Meth-Res

(10_>)

A15097

8th

4th

Sai. enteritidis

(10-)

A9531

0.25

0.13

E. coli Juhl

(10-)

A15119

0.5

1

E. coli

(10-)

A9675

2nd

2nd

K. pneumoniae

(10-)

A9977

0.5

0.5

K. pneumoniae

(10-)

A15130

1

1

Pr. Mirabilis

(10-)

A9900

0.5

0.5

Pr. Morganii

(10-)

A15153

32

32

Ps. Aeruginosa

(10-)

A9843A

> 125

> 125

Ser. marcescens

(10-)

A20019

> 125

> 125

Ent. cloacae

(10-)

A9656

> 125

> 125

Ent. cloacae

(10-)

A9657

0.5

0.5

Ent. cloacae

(10-)

A9659

32

32

* 45% AAB + 5% serum + 50% named broth ** Dilution of overnight broth 'Adjusted to a 79.5% content of BL-S640. The numerical values are thus reduced, i. H. improved. This correction was also made in the tests below or in the alternatives, and a greater amount of weight was used to achieve an equivalent dosage.

Blood levels in mice after IM administration of 10 mg / kg body weight

Urine decrease after IM administration of 50 and 10 mg / kg at

Rats

Association No. of mice

Blood level (mg / ml)

Hours after administration 0.25 0.5 l 1.5

BL-S1052 16 BL-S640 32

15.1 15.7

15.1

13.2

11.5 9.5

7.8 6.8

Compound dose number of 45 (mg / kg) rats

Percent recovered dose administered hours after administration 0-6 6-24 0-24

The compounds were prepared in 0.01% phosphate buffer.

BL-S640 was used as the standard for all connections.

Blood levels in mice after PO administration of 100 mg / kg body weight

BL-S1052

50

4th

40.3

2.1

42.4

10th

3rd

21.7

2.4

24.1

BL-S640

50

7

44.5

2.7

47.2

10th

7

24.3

1.4

25.7

connection

No. of

Blood spouts (ms / ml)

Mice

Hours after administration

0.5 1 2

3.5

BL-S 1052

16

45.2 48.1 31.9

14.4

BL-S640

32

53.4 45.4 27.2

10.7

The compounds were prepared in Tween-CMC. BL-S640 was used as the standard for all connections.

The compounds were prepared in 0.0 Kf phosphate buffer. BL-S640 was used as the standard for all Verbin-55 fertilizers.

Paper chromatograms were prepared with rat urine collected between 0 and 2 and between 2 and 4 hours following IM administration (intramuscularly) of BL-S1052 and BL-S640, for the determination of antibiotic active metabolites 60 using systematic declining chromatography No. 9 (butyl acetate: n-butanol: acetic acid: HiO = 80: 15: 40: 24). Two identically located spots were observed in all cases, except that from the standard, which was not administered to the rats and gave a single identical spot for each of BL-S640 and BL-S 1052. This indicated complete hydrolysis of the derivative to the parent compound BL-S640 and its suspected metabolite.

629 500

14

Example 4

Connections of the following formulas were created:

• n N

—1 Jk

TVT

N H

-s-cj

N-H

•and

CH, 0

3rd

ho jf 0-

H

i-

\ / »

hn.

0

II

• c »

Po cVS ^ ':

i?

N

COOIIa by substituting an equimolar weight of the corresponding amphoteric cephalosporin for the cefatrizine according to the procedure of Example 3. HO

Example 5

There are compounds of the following formulas

■ Oil 11

X — J HNv.

C-h

[4th

0

No. ch2-s

■ N-

N

CH ^

COONa

COONa

15

629 500

H

0

1

II

—C—-

—C

1

1

/ N

C.

A

-H

jÇr

N-

CH2-S

ÏÏ-CH ^

And

COONa

N-C ^

COOha.

prepared by substituting an equimolar weight Example 6

of the corresponding amphoteric cephalosporin for the cefatric 30 The corresponding compounds are tin according to the method of Example 3.

H 0

629 500

16

CH, 0

5 \ H

generated by substituting an equimolar weight of the corresponding amphoteric cephalosporin for the cefatrizine in the method of Example 3.

15 Example 7

Reaction product of BL-S640 and 5-formyl-2-furansulfonic acid sodium salt (BL-S1027) of the formula rs-C3

1) There is a slurry of 9.0 g of 5-formyl-2-furansulfonic acid sodium salt of the formula 30

6) Oral and intramuscular rat blood levels correspond to that of BL-S640.

S03Na)

35

40

45

prepared in 75 ml of water at 60 ° C.

2) 20 g (1 equivalent) of a 1,2-propylene glycol adduct BL-S640 is sprayed onto the sludge with rapid stirring within 10 min while simultaneously adjusting the pH to 6-7.0.

3) The solution is then stirred at 55-60 ° C, pH 6-7.0 for 10 minutes and then cooled to 4-8 ° C. A small amount of precipitation is formed.

4) The mixture is then stirred at 4-8 ° C for 15 min and then filtered.

5) The filtrate is added to a liter of rapidly stirred isopropanol at 5 minute intervals. A heavier precipitate is formed and the mixture is slurried for 5 min.

6) The solids are filtered, washed with 200 ml of isopropanol and dried in vacuo at 56 ° C. for 24 h.

7) The solids are dissolved in 60 ml of water at 4-6 ° C, 55 a small amount of insoluble particles being removed by filtration. The filtrate is lyophilized for 24 h. The yield is 23 g; the bio-yield is 89%.

'N-t

H

Properties of BL-S1027

1) Bio-examination as BL-S640 = 724 mcg / mg (theory = 750 mcg / mg).

2) Solubility = greater than 500 mg / ml.

3) IR-NMR: corresponds to the structure. At 70 mg / ml the compound is present as an 80% cyclic and 20% non-cyclic derivative.

4) It is found by means of paper strip and liquid chromatography that the product is present as a water solution at 0.2 and 1.0 mg / ml as free BL-S640.

5) MIC bacterial spectrum corresponds to that of BL-S640.

7) A solution of this product in water at a concentration of 25.00 mcg / ml was stable, had full bioactivity for at least 3 days at room temperature and for 14 days at 6 ° C in a cold room and thus corresponded to a solution that was suitable is for oral use.

Analvtical data:

Found

Based on dry weight

theory

% H-> 0

% C

5.14

42.1

43.0

% H

40.15

2.9

2.96

% N

3.10

May 13

13.1

% S

12.39

14.35

14.95

Ashes as na

13.6

6.16

6.8 for Di-Na

5.86

17th

629 5

Antibiotic activity in the nutrient broth

Organism MIC (mcg / ml)

Sodium salt Cpd of Cap. V

BL-S640-

Cephalothin

St. pneumoniae *

(10 - ') **

A9585

0.03

0.13

0.03

St. pyogenes *

(10 - *) **

A9604

0.03

0.06

0.03

S. aureus Smith

(10-)

A9537

0.27

0.5

0.13

S. aureus + 50% serum

(10-)

A9537

2nd

4th

0.5

S. aureus BX1633

(10-)

A9606

0.5

0.5

0.13

S. aureus BX1633

(10-)

A9606

4th

4th

0.25

S. aureus Meth-Res

(10 ")

A15097

4-8

8-16

1-16

Sai. enteritidis

(io-)

A9531

0.25

0.25

0.25

E. coli Juhl

(10-)

A15119

1

1

16

E. coli

(10-)

A9675

- 4th

4th

32

K. pneumoniae

(10-)

A9977

0.5

0.5

1

K. pneumoniae

(10-)

A15130

1

2nd

32

Pr. Mirabilis

(10-)

A9900

0.5

0.5

1

Pr. Morganii

(10-)

A15153

32

63

> 125

Ps. Aeruginosa

(10-)

A9843A

> 125

> 125

> 125

Ser. marcescens

(10-)

A20019

> 125

> 125

> 125

Ent. cloacae

(10-)

A9656

> 125

> 125

> 125

Ent. cloacae

(10-)

A9657

1

1

8th

Ent. cloacae

(10-)

A9659

32

63

> 125

* 45% A AB + 5% serum + 50% above broth ** Dilution of broth culture kept overnight 'These numbers were calculated back based on where only 65% BL-S640 was present in the sample; in other words the numerical values have been reduced, i. H. improved.

- The chemical name for BL-S640 has already been given above.

Blood levels in mice after oral administration of 100 mg /

kg body weight 35

connection

Blood level ([ig / ml)

Hours after administration

0.5

1

2nd

3.5

Connection of Example 7

45

36.6

20.4

10.4

BL-S6401

50.5

39

24.5

9.6

Connection of Example 7

54.6

53.5

27.6

8.2

BL-SÓ40 '

57.4

47

29.5

10.9

be stored for at least 24 hours with no significant loss of activity.

Blood levels in mice after intramuscular administration of 10 mg / kg body weight

connection

Hours after administration

45

These compounds were prepared in tween carboxymethyl cellulose and 8 mice were used for each compound.

1 BL-S640 was used as standard, the chemical name of which was given above.

BL-S1027 is the water-soluble derivative formed when B1-S640 is reacted with furfural 50 sodium sulfonate. The bio strength is 724 mcg / mg of BL-S640 effectiveness. Aqueous solutions with 250.25 and 10 mg BL-S640 activity / ml can be used as BL-S1027 with cooling at 4 ° C or at room temperature.

0.25

0.5

1

1.5

Connection of Example 7

16.1 14.6

14.8 15.4

11.9 11.7

8th

8.3

BL-S6401

16.3

13.5

9.4

6.3

1 It was used as a standard BL-S640, the chemical name of which is given above.

The compounds were dissolved in 0.01% phosphate buffer.

Example 8

Reaction product BL-S1048 of the formula

H

0

H

I.

629 500

18th

obtained from BL-S640 and o-formylphenoxyacetic acid. 1) 375 mg of o-formylphenoxyacetic acid

CHO

was slurried in 10 ml of water and 4N NaOH was added to pH 7.0 with rapid stirring. A solution was obtained.

2) 2 g (1 equivalent) of BL-S640-methanol adduct were added with rapid stirring at 10 minute intervals and simultaneous addition of 4N NaOH to a pH of 6-6.5. A solution or a fast solution was formed.

3) The solution was filtered and kept at ambient temperature for 0.5 h.

4) The solution was lyophilized for 24 hours and the product was obtained as a solid.

Properties of the solid product

1) IR-NMR (70 mg / ml) corresponded to the structure, the triazole and | 3-lactam ring is intact.

5 2) Pupierstreifen- and liquid chromatography at 0.2 and 1 mg / ml showed a BL-S640 completely free of aldehyde.

Analytical data OK

Found

Based on dry weight

theory

% Water

4.06

_

_

% C

47.15

49.2

50.1

% H

3.79

3.5

3.86

% N

11.44

11.93

13.0

% S

8.87

9.27

9.91

Ashes as na

5.55

5.78

6.47

3) Solubility in water => 400 mg / ml.

Antibiotic spectrum of the nutrient broth

Organism MIC (mcg / ml)

BL-S640

BL-S 1027 '

BL-S10482

S. pneumoniae *

(IO "3) **

A9585

0.06

0.03

0.06

St. pyogenes *

(10 - ') **

A9604

0.03

0.03

0.03

S. aureus Smith

(10-)

A9537

0.13

0.25

0.13

S. aureus + 50% serum

(10-)

A9537

2nd

2nd

2nd

S. aureus BX1633

(io-1)

A9606

0.25

0.5

0.25

S. aureus BX1633

(io-2)

A9606

4th

4th

2nd

S. aureus Meth-Res

(IO-3)

A15097

8th

4th

4th

Sai. enteritidis

(10-)

A9531

0.25

0.25

0.13

E. coli Juhl

(10-)

A15119

0.5

1

0.5

E. coli

(10-)

A9675

2nd

4th

2nd

K. pneumoniae

(io-)

A9977

0.5

0.5

0.5

K. pneumoniae

(io-)

A15130

1

1

1

Pr. Mirabilis

(io-)

A9900

0.5

0.5

0.5

Pr. Morganii

(io-)

A15153

32

32

32

Ps. Aeruginosa

(io-)

A9843A

> 125

> 125

> 125

Ser. marcescens

(io-)

A20019

> 125

125

> 125

Ent. cloacae

(io-)

A9656

> 125

> 125

> 125

Ent. cloacae

(io-)

A9657

0.5

1

0.5

Ent. cloacae

(10-)

A9659

32

32

32

* 45% AAB + 5% serum + 50% broth mentioned above ** Dilution of culture broth left overnight. 'Set to 68% BL-S640 content - Set to 67% BL-S640 content

The numerical values were thus reduced, i. H. improved. This correction was also made in the test trials listed below or, according to an alternative, a larger weight was used to provide equivalent dose levels.

Blood levels in mice after IM administration of 10 mg / kg body weight. Mice after PO administration of 100 mg / kg body weight

connection

No. of the mice

Blood level (mcg / ml)

Hours after administration 0.25 0.5 1

1.5

60

connection

No. of the mice

Blood level (mcg'ml)

Hours after administration 0.5 1 2

3.5

BL-S1027

16

15.4

15.1

11.8

8.2

BL-S 1027

16

49.8

45.1

24th

9.3

BL-S 1048

16

14.8

14.1

10.3

7

65 BL-S 1048

16

46.5

44.6

28.1

12, p

BL-S640

32

15.7

13.2

9.5

6.8

BL-S640

32

53.4

45.4

27.2

10.7

The compounds were generated in residual phosphate buffer. BL- The compounds were prepared in Tween-MCM. As standard for S640 was used as standard for all connections. all connections were used BL-S640.

19th

629 500

Urine collection after IM administration of 50 and 10 mg / kg

Rats

Compound dose number of (mg / kg) rats

Recurrence of the administered dose in '/ p hours after administration uim 0-6 6-24 (> -24

BL-S 1027

50

4th

39.6

2.6

42.2

10th

4th

30.1

1.7

31.8

BL-S1048

50

4th

32.3

2.4

34.7

10th

4th

21.7

2nd

23.7

BL-S640

50

7

44.5

2.7

47.2

10th

7

24.3

1.4

25.7

The compounds were prepared in 0.0 1% phosphate buffer. BL-S640 served as the standard for all connections.

H

Paper chromatograms were performed on rat urine collected between 0 and 2 and between 2 and 4 hours after IM administration, supplement 0 mgm / kl and at 50 mgm / kg, of BL-S1027,1048 and 640, for the purpose of monitoring -5 white of antibiotic active metabolites using the descending chromatography with system No. 9 and using butyl acetate: n-butanoI: glacial acetic acid: water in the ratio 80: 15: 40: 24. Two identical spots were observed in all cases other than the standard, i.e. H. not originating from the animal, which had a single identical spot. This indicates complete hydrolysis of the two derivatives to the parent compound BL-S640.

Example 9

15 Preparation of the reaction product of BL-S640 and o-benzaldehyde sulfonic acid

I?

; -F S

T

H

1) 870 mg of sodium salt of o-benzaldehyde 30 4) Bioassay = 700 mcg / mg. sulfonic acid dissolved in 10 ml of water.

2) 2 g of BL-S640-methanol adduct (1 equivalent) were added to this solution with rapid stirring within 10 min with simultaneous addition of 4N NaOH to a pH of 6.5. A solution was obtained.

3) The solution obtained was filtered, left to stand at room temperature for 1 h and lyophilized for 24 h,

and the desired product was obtained as a solid.

5) IF-NMR spectra show an intact β-lactam and triazole part and the compound does not appear to be a cyclic adduct.

Properties of the product

1) IR-NMR spectra are well defined and correspond to the structure.

2) Solubility in water is greater than 400 mg / ml.

3) Analytical data

Found based on dry matter theory

40

6) Paper strip chromatography showed that there appears to be a double zone of the Rf of BL-S1640.

7) Liquid chromatography:

Time in hours

% of free BL-S640

45

64.0 70.5 88.0

% Water

3.49

-

_

% C

41.88

43.4

43.3

% H

3.41

3.3

3.25

% N

11.53

12.1

12.35

% S

13.74

14.25

14.24

Ashes as na

5.45

5.63

6.77

50

8) Solubility in water is greater than 400 mg / ml.

9) This product is called BL-S 1055.

organism

55

Antibiotic spectrum in nutrient broth MIC (mcg / ml)

BL-S 1055 '

BL-S640

S. pneumoniae *

(10 ') **

A9585

0.06

0.03

St. pyogenes *

(10 - ') **

A9604

0.016

0.03

S. aureus Smith

(10 ')

A9537

0.13

0.13

S. aureus + 50% serum

(io- ')

A9537

2nd

2nd

S. aureus BX1633

(10-0

A9606

0.5

0.13

S. aureus BX1633

(10 -)

A9606

2nd

2nd

S. aureus Meth-Res

(10 ')

A15097

4th

2nd

Sai. enteritidis

(10 years)

A9531

0.35

0.13

E. coli Juhl

(10 ')

A15119

1

0.5

629 500 20

Antibiotic spectrum in nutrient broth organism MIC * (mcg / ml)

BL-S 1055 'BL-SMO

E. coli

(10 0

A9675

4th

2nd

K. pneumoniae

(10 ")

A9977

0.5

0.5

K. pneumoniae

(10-)

A15130

1

1

Pr. Mirabilis

(io-)

A9900

0.5

0.5

Pr. Morganii

(10 o

A15153

32

16

Ps. Aeruginosa

(10-y)

A9843A

> 125

. > 125

Ser. marcescens

(10- ')

A20019

> 125

63

Ent. cloacae

(10-0

A9656

> 125

63

Ent. cloacae

(lo-o

A9657

0.5

0.5

Ent. cloacae

(10-0

• "A9659

63

63

* 45% AAB + 5% serum + 50% broth as stated above "Dilution of culture broth held overnight" Set to 65% content of BL-S640. The numerical values were thus reduced, ie improved. This correction was also carried out in the tests listed below carried out or according to an alternative, a larger weight was used to obtain an equivalent dosage.

Blood levels of mice after IM administration of 10 mg / kg urine after IM administration of 50 mg / kg in rats

body weight

Compound no. Of recovery in% of.

Compound blood level (mcg / ml) dose administered to 25 rats

Hours after administration hours after administration 0.25 0.5 1 1.5 0-6 6-24 0-24

BL-S1055 '10.3 9.96 7.6 6.5 BL-S10551 3 36.6 2.4 39

BL-S640 12.7 11.5 8 5.7 BL-S640 4 44.2 3.3 47.5

30th

The compounds were generated in 0.01% phosphate buffer. The compounds were generated in 0.01% phosphate buffer. Values correspond to the average of 3 attempts. 'BL-S640 served as the standard.

'BL-S640 was used as standard.

Blood levels of mice after PO administration of 100 mg / kg

Body weight 35

Paper chromatograms were performed using rat

Compound blood levels (mcg / ml) only collected between 0 and 2 and between 2 and 4 hours after

Hours after administration ^ _ IM administration of BL-S1055 and 640, namely for the

: Detection of antibiotic active metabolites using

BL-S1055 '38.2 35.7 22.0 11.3 40 descending chromatography with System No. 9, the

BL-S640 48.7 42.4 24.0 10.1 is with butyl acetate: n-butanol: glacial acetic acid: water in the ratio 80: 15: 40: 24. A single identical spot was observed in all cases. This shows a complete hydro

'BL-S640 lysed the derivatives to the parent compound BL-S640 as standard.

45 x

Comparative oral PD5o values of BL-S640 derivatives in mice

organism

PD51, (mg / kg / load) *

BU-S640

BL-S 1027

BL-S 1048

BL-1055

Cephalexin

E. coli

A15119

1.4

-

3.1

1.8

13

4.1

-

1.6

1.6

13

P. mirabilis

A9900

7.2

6.3

4.7

8.2

66

9.6

2.7

3.6

8.2

55

P. mirabilis

A9707

2.4

1.8

2.4

1

50

K. pneumoniae

A9977

1.6

-

0.7

1.6

43

S. pyogenes

A9604

0.4

-

0.2

0.4

11

"Mice were treated at 1 and 3.5 h after excitation

Comparative IM PD ^ values of BL-S640 derivatives in mice organism PD. "(Mg / kg / treatment) *

: BL-S640 BL-S 1027 BL-SI048 BL-1055 Cephalexin

P. mirabilis A9900 - - - _ _

• 1.6 1.6 1.6 1.6 50

21st

629 500

Example 10

Production of BL-S 1027 of the following structural formula

0

II

- c

1

N

by reacting BL-S640 and 5-formyl-2-furansulfonic acid

\ r = / hw

(D, L)

rf.

W

H2 °

COONa

SO ^ Well? _

1) 9 g of the sodium salt of 5-formyl-2-furansulfonic acid of the formula were filtered and lyophilized directly as described in steps 8-9 above.

(oh4jL

S0oNa stirred rapidly in 75 ml of water at 60-65 ° C, whereby most of the solid dissolved.

2) 20 g of BL-S640-propylene glycol adduct were sprayed onto this solution with rapid stirring within 10 min, while the pH was adjusted to 6.5-7.0 with 40% NaOH, the pH being 7. Should not exceed 2. A solution or a fast solution was formed.

3) The solution is stirred at pH 6.5-7.0 at 55-60 ° C for 15 minutes and then cooled to 4-8 ° C. Stirring at 4-8 ° C is continued for 10 min.

4) It is then filtered to remove small amounts of the insoluble.

• 5) The filtrate (see footnote 1) is then added to 11 isopropanol with rapid stirring. An amorphous precipitate is formed; stirring is continued for 5 minutes

6) The solid formed is filtered off, washed with 100 ml of isopropanol and dried in vacuo at 50-56 ° C. for 18 hours.

7) The solid mentioned is dissolved in 60 ml of water and cooled to 4-6 ° C. A very small amount of precipitate can form and can be filtered off.

8) This filtrate is filtered through suitable filters at ambient temperature in order to remove particles, febrile substances and bacteria.

Levels 7 and 8 should take place within 4 hours.

9) It is then sterile lyophilized for 48 hours and then heated at 50-56 ° C under vacuum for 18-24 hours. The yield of BL-S1027 is approximately 22-24 g.

Note 1 from Step 5: If 18.7 g of BL-S640-methanol adduct is substituted for the 20 g of BL-S640-propylene glycol adduct used, the filtrate can be cooled to 4-6 ° C, then

20th

Properties of BL-S1027

1) Bio-test resulted in 724 mcg / mg.

2) Bio yield is 85-95%.

3) IR-NMR spectra corresponded to the structure and there was about 80% cyclic, 20% non-cyclic structure in D20

found at 70 mg / ml.

Finally 0.05 mol of isopropanol was found.

4) Paper strip chromatography at 0.2 mg / ml gave 100% 30 BL-S640 and no sign of the presence of a derivative.

5) Liquid chromatography 1 mg / ml in pH 7 phosphate buffer

. showed 85% free BL-S640 at 0 time, 94% free BL-S640 after 0.5 h and 100% free BL-S640 after 1.0 h.

6) Water solubility is greater than 400 mg / ml. 35 7) Analytical data

40

45

Found

Based on dry basis

theory

% HiO KF '

5.14

-

_

% C

40.15

42.1,

43.0

% H

3.10

2.9

2.96

% N

12.39

13.05

13.1

% S

13.6

14.35

14.95

Ashes as na

5.86 '

6.16

6.5

8) Blood levels in mice, namely levels during oral and intramuscular treatment, roughly equivalent to BL-S640-

50 propylene glycol adduct. BL-S640 appears in the urine without evidence of the derivative.

9) a) Testing the stability of the solids showed a loss of less than 10% during 1 month at 56 ° C.

b) When testing the stability of solutions, it was found that a loss of less than 10% occurs at 250.25 and 10 mg / ml BL-S640 effectiveness for at least 24 h at room temperature.

2 TEA +

629 500

22

° 18H18H6 ° 5S2 U62-50

OHC

n

+ KEH

SO ^ Na 3

h0 "(o) —t

N f H-N-

0

II

CH N-I

f oh2-S

H

1 1?

G

<5

_! . C02K

SO_, Na 5

C23H17ïï608S2NaX

35

Materials:

1 g (0.00217 mol) of BL-S640 429 mg (0.0217 mol) of the sodium salt of 5-furfural sulfone

acid 40

217 mg (0.0434 mol) triethylamine 1.2 g (0.0434 mol) potassium 2-ethylhexanoate (KEH)

Procedure: 27 mg of triethylamine (TEA) and 429 mg of 5-45 furfural sulfonic acid were added to a suspension of 1 g of BL-S640 in 8 ml of methanol. The mixture was heated to reflux, and the solids soon dissolved. The solution was then heated for 3 minutes and 1.2 g KEH was added. A yellow salt separated, it was collected and washed with methanol. This salt was then dried over P2Os 50 in a desiccator and the yield was 800 mg. Analysis for C23Hi7Nfi08SiNaK-4H ^ 0 calcd .: C 37.54 H 3.41 N 11.42 found: C 37.30 H 3.31 N 10.18

F greater than 140 ° C (dec.). 55

The same reaction as described above was carried out, except that sodium 2-ethylhexanoate was used instead of KEH. The salt did not precipitate from methanol, so the solution was filtered and the filtrate was diluted with isopropyl alcohol. The precipitate formed was collected and washed with isopropyl alcohol. The salt was hygroscopic, so it had to be vacuum dried over P2Os. The yield was 800 mg, F> 140 ° C dec.

Analysis for Ci3HI7N60 «S.ìNa calc .: C 42.65 'H 2.78 N 13.65« 5

Found: C 41.03 H 3.94 N 10.54

The sample was soluble in water and the solution showed a pH of 6.5. '

A paper strip test was carried out in buffer and in human serum. The sample showed only one spot corresponding to BL-S640. Both samples had a solubility of> 100 mg / ml at pH 6.5.

Paper strip chromatography showed that the above compound was completely hydrolyzed to BL-S640 in pooled human serum at pH 7 and 0.1M phosphate buffer at pH 7. This paperstrip bioautograph was carried out in the following manner. Samples were dissolved in 0, IM pH 7 phosphate buffer and pH 7 human serum at 500 y / ml at the concentration of 500 y / ml. A 10 (xl sample of BL-S640 and the above product were spotted on a 12.5 mm strip of What # 1 paper and placed in a system of butyl acetate 80, n-butanol 15, at 27 ° C for 16 h. Acetic acid 40 and water 24 were developed and the strips were then air dried and applied to an agar plate inoculated with B. subtilis ATCC 6633. A single zone of Rf 0 was seen at 30 min, 1 hour, 3 h and 6 h intervals , 3 for all samples.

High pressure liquid chromatography at a concentration of 1 mg / ml showed complete hydrolysis to the parent BL-S640 compound after half an hour. Samples were chromatographed on an nBonda-pak C18 (Waters Associates) column using a 0.01M sodium acetate pH 4 / methanol 8/2 mobile phase at a flow rate of 0.52 ml / min. The detection was carried out by UV at 354 (.im. Samples were dissolved at a concentration of 1 mg / ml in the mobile phase, 2 n were injected and BL-S640 was eluted at 18 min under aldehyde at 6 min. The quantification of BL -S640 and 5-formylfuransulfonic acid were made using p-toluenesulfonic acid as an internal standard.

23

629 500

Example 12

There were compounds of the formula

COONa, in which R2 is phenyl-2-sulfonic acid, 4-methoxyphenyl-3-, in which the 5-formyl-2-furansulfonic acid is

sulfonic acid, 4-hydroxyphenyl-3-sulfonic acid, 2-carboxymethers of Example 7 by an equimolar weight of the corresponding

xyphenyl, 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxyphe- xing aldehyde of the formula R2-CHO was replaced, nyl, 4- (2'-carboxy) vinylphenyl, carboxyl, 2-carboxyphenyl, 3-

Carboxyphenyl, methanesulfonic acid or methane disulfonic acid

Example 13 There were compounds of the formulas

COONa and where R2 is ch, 0

? '

HO

COONa

■ 04

ML

0

II

• c \

c l

R

0-CH2-S "C

a 0 I H

in !

N

I.

COOIÏa

S03Na stands, manufactured, namely the amphoteric cephalosporins for that in Example 7

used cefatrizine.

55

by substituting an equimolar weight of the corresponding

Example 14 There were compounds of the formulas

COONa

629 500

24th

"OK

No

CH2-S

-n-cel I 3

: N

COONa and where R2 for-

'0,

GH,

-b-j

HN.

^ C-K

»2

R

n-

ch2-s

COONa

• N-CH-, l 3 "N

N-N-CH ^

I I

chg-s-il

COOfta

■ S03Na stands, generated by 40 amphoteric cephalosporins for cefatrizine in the process of

Example 7.

Example 15

Substitute an equimolar weight of the corresponding compounds of the formulas

-p ^ l_ JPM

Ii n ^ n I

COONa CH,

j5

oJ-S ^ -ch2-S klK N,

COONa CH,

5

and wherein R2

0

I I

25th

629 500

CH,

, ~ S

COONa

W

I.

CH-

N I

N

2 0 '

J-N ^^ - CH „-S JC int /

N

i

✓ N

COONa

N '

I.

CH-;

S03Na is made by 25 amphoteric cephalosporins for that in the procedure of Example 7

used cefatrizine.

Example 16

Substitute an equimolar weight of the corresponding compounds of the formulas

30th

HO HO

V = / TTW

c I

• R

^ JXÌch2-SJT7

2 0 ^ Y '■ N

COONa H

-N 11

COONa

2 0 'I H

R

N I

N

I.

COONa and prepared, wherein R: phenyl-2-sulfonic acid. 4-methoxyphenyl-3-xyphenyl, 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxyphe-sulfonic acid, 4-hydroxyphenyl-3-sulfonic acid. 2-carboxymethonyl, 4- (2'-carboxy) vinylphenyl. Carboxyl, 2-carboxyphenyl, 3-

629 500

26

Carboxyphenyl, methanesulfonic acid or methanedisulfonic acid-containing aldehyde of the formula R2-CHO has been replaced, specifically by the 5-formyl-2-furansulfonic acid in the process of Example 7 having an equimolar weight of the corresponding Example 17

jj q Connections of the formulas ■

V il TT!

COONa

- HO

COONa and

COONa is produced, in which R2 is phenyl-2-sulfonic acid, 4-methoxyphenyl-3-sulfonic acid, 4-hydroxyphenyl-3-sulfonic acid, 2-carboxymethoxy-xyphenyl, 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxyphenyl, 4- ( 2'-carboxy) vinylphenyl, carboxyl, 2-carboxyphenyl, 3-carboxyphenyl, methanesulfonic acid or methane disulfonic acid, respectively, by replacing those in the process of Example 7

H

\ 'w

HN>

COONa used 5-formyl-2-furansulfonic acid due to an equimola-55 res weight of the corresponding aldehyde of the formula R2-CHO.

Example 18 Compounds of the formulas fiO

C.

1

R

CH,

a y— ■

, -s-c

N

II

COONa

N "

I.

CH.

N

27th

629 500

H0 H

and

CH-0

■ * \ H

f

CH,

, -S-X

COONa

N-

I.

CH-

ì?

✓N

generated in which R2Phenyl-2-sulfonic acid, 4-methoxyphenyI-3-sulfonic acid, 4-hydroxyphenyl-3-sulfonic acid, 2-carboxymethoxy-xyphenyl, 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxyphenyl, 4- (2 ' Carboxy) vinylphenyl, carboxyl, 2-carboxyphenyl, 3-carboxyphenyl, methanesulfonic acid or methanedisulfonic acid, respectively, by replacing those in the process of Example 7

40

used 5-formyl-2-furansulfonic acid by an equimolar weight of the corresponding aldehyde of the formula R2-CHO.

Example 19

Preparation of the sodium salt of BL-S1057 of the formula

H

• 2h20

by reacting BL-S643 and 2-furaldehyde.

1) Prepare a slurry of 1 g BL-S643-3HiO in 10 ml water at 40-45 ° C.

2) 0.25 g (1.25 equivalents) of 2-furalde-hyd are added to this.

3) It is then added to pH 7-7.5 in rapid stirring in sodium hydroxide solution and a solution or fast solution is obtained in less than 5 minutes.

4) This solution is cooled to 22-24 ° C and treated with a suitable filter to remove particles, bacteria and fever-producing substances.

5) This solution is then lyophilized under sterile conditions for 48 h and BL-S1057 is obtained as a powder; this substance can also be obtained by precipitating stage 4 sterile solution 60 with 15-20 volumes of sterile isopropanol.

Properties of BL-S 1057

a) Bioassay using cefatrizine as standard 65 gave 759 mcg / mg.

b) IR spectrum showed an intact, well-defined β-lactam ring.

c) NMR gave a well-defined corresponding structure that

629 500

28

about 75% corresponded to the cyclic adduct (60 mg / ml in D20). d) Solubility is greater than 400 mg / ml in water.

4) Paper strip chromatography primarily showed a zone at Rf of BL-S643 at 37 ° C at 0 and 1 h and at a concentration of 0.2 ms / ml.

at a concentration of 1 mg / ml.

g) F at 180 ° C (shrinkage) at 200 ° C decomposition.

Analytical data

Found

Related to

Drying

Base

theory

% H, 0 KF

5.18

-

_

% C

45.91

48.4

48.4

% H

4.05

3.7

3.37

% N

10.81

11.4

11.8

% S

14.92

15.73

16.48

Ashes as na

'3.84

4.03

3.88

Example 20 Generation of BL-S1058 of Formula n n jj-ch-

• 5H2O

coon_

by reacting BL-S643 and sodium 5-formyl-2-furan-sulfonic acid.

1) There is a slurry of 0.45 g of 5-formyl-2-furansulfonic sodium salt

SOglTa

30th

35

SO, Na b) Investigations with IR and NMR showed a well-defined, expected structure with an intact ß-lactam and 3-side chain and a 90% cyclic adduct was found at about 60 mg / ml in D20.

c) Using paper strip chromatography, a zone was primarily found for Rf of BL-S643 at 0 and 1 h at 37 ° C and with a concentration of 0.2 mg / ml. There is evidence of a second zone.

d) Liquid chromatography with 1 mg / ml in pH 6 buffer.

prepared in 10ml water at 40-45 ° C. You get a solution or a fast solution.

2) Add BL-S643-3H2O within 10min under 40 rapid stirring and at the same time add IN pH 7-7.5 in sodium hydroxide solution. A solution or fast solution is formed within 5 minutes.

3) The solution is cooled to 20-23 ° C and filtered through suitable filters to remove particles, bacteria and pyrogenic substances 43.

4) It is then lyophilized for 48 hours and the powder obtained is the above-mentioned BL-SI058; Sterile BL-S1058 can also be obtained from the step 3 solution by precipitation from 15-20 volumes of sterile isopropanol. 50

Properties of BL-S1058

a) Bioassay using cefatrizine as standard gave 685 mcg / mg.

Time in hours

% hydrolyzed

0

25.4

1

46.5

2nd

59.0

Analytical data

Found

Dry

theory

Base

% H, 0 KF

10.17

-

-

% C

36.5

40.5

41.6

% H

2.89

2.4

2.74

% N

8.52

9.49

10.01

% S

17.09

18.98

18.6

Ashes as na

7.04

7.93

6.63

29

629 500

organism

Antibiotic spectra of BL-S643 derivatives of the nutrient broth

MIC (mcg / ml)

BL-S10571

BL-S 1058-

BL-S643

S. pneumoniae *

(10 ') **

A9585

0.016

0.016

0.03

St. pyogenes *

(io y

A9604

0.008

0.008

0.016

S. aureus Smith

(io-4)

A9537

0.5

0.25

0.5

S. aureus + 50% serum

(10 "0

A9537

8th

8th •

8th

S. aureus BX1633

(10- ')

A9606

1

0.5

1

S. aureus BX1633

(10-0

A9606

4th

4th

4th

S. aureus Meth-Res

(10-0

A15097

2nd

2nd

4th

Sai. enteritidis

(10-0

A9531

0.25

0.13

0.25

E. coli Juhl

(10-0

A15119

1

1

1

E. coli

(lo-O

A9675

4th

2nd

4th

K. pneumoniae

(lo-o

A9977

0.5

0.5

0.5

K. pneumoniae

(lo-o

A15130

4th

2nd

4th

Pr. Mirabilis

(lo-o

A9900

0.5

0.5

0.5

Pr. Morganii

(lo-o

A15153

16

16

16

Ps. Aeruginosa

(io-)

A9843A

> 125

> 125

> 125

Ser. marcescens

(lo-o

A20019

125

125

125

Ent. cloacae

(lo-o

A9656

> 125

> 125

> 125

Ent. cloacae

(lo-o

A9657

0.5

0.5

4th

Ent. cloacae

(lo-o

A9659

16

16

32

R =

JJL

SO ^ Na

* 45% A AB + serum + 50% broth mentioned above

** Dilution of overnight culture broth 'Adjusted to 80% BL-S643 2 Adjusted to 59% BL-S643

The numerical values were thus reduced, i. H. improved. This correction was also made in the tests below or in the alternative, using a larger weight in order to achieve an equivalent dosage.

N-

■ N

-S — II YY-

CH-

COONa

Paper chromatograms were prepared using rat urine collected between 0 and 2 and between 2 and 4 hours after IM administration of BL-S1057,1058 and 643 for the detection of antibiotic active metabolites using decreasing chromatography with the System No. 9, i.e. H. with butyl acetate: n-butanol: glacial acetic acid

50

rerHjO in the ratio of 80: 15: 40: 24. A single, identical spot was observed in all cases. This indicates complete hydrolysis of the two derivatives to the parent compound BL-S643.

Example 21 Compounds of formula

N,, N

-s ji-

CH ^

COONa produces, wherein R2 is phenyl-2-sulfonic acid, 4-methoxyphenyl-3-sulfonic acid, 4-hydroxyphenyl-3-sulfonic acid, 2-carboxymethoxy-xyphenyl. 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxyphenyl, 4- (2'-carboxy) vinyl Ipheny 1. carboxyl, 2-carboxyphenyl, 3-

65 carboxyphenyl, methanesulfonic acid or methanedisulfonic acid is prepared by replacing the 5-formyl-2-furansulfonic acid in the process of Example 20 with an equimolar weight of the corresponding aldehyde of the formula R: -CHO.

629 500

30th

Example 22

There were connections of the formulas

N: N

n n ch „-s —Ji— ch5

© the ch5 ° x h 0

i 'n ho - // \ vc c

"== / hn ^ n n

-N ^ ch2-S —11 ^ ^ JJ — CH ^

where R2 is -Q or "nf irso? times;

generated by substitution of an equimolar weight of the corresponding amphoteric cephalosporin for the cefaparole in procedures of Examples 19 and 20.

31

629 500

Example 23

There were compounds of the formula

T

rtl

'Hv -CH0 ~ S-C

COONa

1f

.N

N

i

CH-,

HO

or where R:

■■ -A ° dcr ~ if ° "ir

SO, Na is by substituting an equimolar weight of the corresponding amphoteric cephalosporin for the cefaparole in the methods of Examples 19 and 20.

629 500

32

Example 24

There were compounds of the formula

HO

H

0

1

II

-C -

- C

1

1

HN ^

HO

H <

* 0: *

N -N

CH0 ~ S-Ii JJ-CH- * 0

COONa

ç c

I 1 /

HN> ^ / N — i f

C-H 1 1

CH-

H <j = V

| a a

H 0

[II

Xh

N. N

CH,

0

CH-,

COONa

N f HN ^ ^ N — i ^

C-H

H2

Y-N

CH.

N jN

»-S — iL JL CH ^ or

COONach, 0

5 \ H 0

H0 ~ Q-M w "^ 77

I.

-N

R

0.

2 0 '

ch2-s

N N

II I

CH-

where R: for

"qor t ìt

COONa is produced by substituting an equimolar SO jNa weight of the corresponding amphoteric cephalosporin for the cefaparole in the methods of Examples 19 and 20.

Example 25 Compounds of formula

COONa

33

629 500

COONa c ecHO '

H

I.

C-I

HN.

C.

t

H'

-C f?

2 <y * N

N or

COONa

ïrQ-

ch,

i3

N-

-N

ch2-s.

COONa is generated by substituting an equimolar in R2 for f | |) or fj j | S03Na weight of the corresponding amphoteric cephalosporin for

|| 11 11 11 40 the cefaparole in the process of Examples 19 and 20.

Example 26 There were compounds of the formulas

N '

ch2-s—

-c-ch, l 3 jn

COONa

COONa

629 500

34

COONa

CH, 0

5 \ H • 0

H0-O-7- "

7 HNv. / N

H — fl-CB,

where R- for

~ f ° ì * fvso '

COONa

25th

is produced, by substituting an equimolar 3Na weight of the corresponding amphoteric cephalosporin for the cefaparole in the method of Examples 19 and 20.

30th

Example 27 There were compounds of the formulas

35

HO-

H

0

1

II

-C-

C.

1

HNV

x yJÏ-

C-H

E2 0

CH ^ S

COONa

COONa

N N

I.

N-CH-j

• r

N N

k N-CH, or N

COONa

35

629 500

^, 0 «.

where R: for - »y Jj or —jj j

■ SO yes

N-

CH2-S

-N

JS-

• r

CH ^

COONa is generated by substituting an equimolar weight of the corresponding amphoteric cephalosporin for 15 the cefaparole in procedures of Examples 19 and 20.

Manufacture of raw materials

Preparation of 7- [Da-amino-a- (p-hydroxyphenyl) aceta-mido] -3- (l, 2,3-triazol-5-ylthiomethyl) -3-cephem-4-carboxylic acid methanol solvate (BL -S640)

6.5 g (11.55 mmol) 7- [Dat-butoxycarbonylamino-a- (p-hydroxyphenyl) acetamido] -3- (1,2,3-triazol-5-ythiomethyl) -3-cephem-4- carboxylic acid was dissolved in 175 ml of 98-100% formic acid under anhydrous conditions. The mixture was stirred at room temperature for 2J4 h. A portion of the solution, 125 ml, was evaporated under reduced pressure to an amber oil. The oil was then azeotroped three times with 70 ml of toluene under reduced pressure. The residue was suspended in 700 ml of an 80:20: H2O-CH3OH solution and stirred for 0.5 h until most of the solid went into solution and then filtered. The filtrate was treated with 1.5 g of "Darko" activated carbon for about 20 minutes. The activated carbon was then filtered through a "Celite" layer, the solution was then freeze-dried in 9 separate 100 ml round-bottom flasks and 2.415 g were obtained. This material was then crystallized in 0.200 g amounts and a total of 0.923 g of 7- [Da-amino-a- (p-hydroxyphenyl) acetamido] -3- (1,2,3-triazole-5-) was obtained. ylthiomethyl) -3-cephem-4-carboxylic acid (BL-S640). NMR met expectations and showed the presence of Vi Mol of CH3OH. Analysis for CISH, RN60 <; S2-H, 0. <A CH3OH:

calc .: C 44.83, H 4.48, N 17.10, S 13.09,

Found: C45.77.44.36, H4.44.4.34, N 16.61.16.52, S 13.01.13.01.

Production of crystalline methanol sulfate from BL-S640

1) 50 g of BL-S1640 was slurried in 250 ml of 95% vol / vol methanol / water, 95% methanol solution at 22-25 ° C.

2) Concentrated hydrochloric acid was then added with rapid stirring to a pH of 1.3-1.5. A solution or a fast solution was formed.

3) This solution was adjusted to a pH of 1.7 with triethylamine.

4) 7.5 g of activated carbon (“Darco G-60”) were then added and the mixture was stirred to form a slurry for half an hour.

5) The activated carbon was then filtered and washed with 75 ml of methanol, which was then added to the filtrate. Steps 2, 3 and 4 should be completed within 5 hours.

6) The combined wash liquors and the filtrate from step 5 were stirred rapidly. Triethylamine was then added to pH 4.5 within 5 min. Crystallization started in about 1-3 minutes. The mixture was then slurried for 1 hour.

7) The crystals were collected by filtration, washed with 100 ml of methanol and dried in vacuo at 56e C for 24 h. The organic yield was 75-90%; the bio-test showed

850-900 mcg / mg; NMR-IR corresponded to one mole of methanol; % H20, KF = 2-4.0.

20th

Production of crystalline 1,2-propylene glycol solvate from BL-S640

1) 25 g of the BL-S640 methanol solvate prepared above was dissolved in 150-200 ml of a solution of 75% vol / vol propylene.

• Glycol water slurried at 20-25 ° C.

2) To this, concentrated hydrochloric acid was added to a pH of 1-1.2, whereby a solution or a fast solution was obtained.

3) Triethylamine (TEA) was added slowly with rapid stirring to pH 1.7-1.8.

4) 5 g of "Darco G-60" were added and the mixture was slurried for half an hour. The carbon was filtered off with slow filtration, suggesting 18.5 cm SS No. 576 paper. The

35 carbon filter cakes were washed with 40 ml of a solution of 75% vol / vol propylene glycol water and the wash water was added to the filtrate. Levels 2, 3 and 4 should be completed within 5 hours.

5) Trimethylamine was then added to a pH of 4.5 within 40 of 10 min to the rapidly stirred filtrate / wash water mixture from stage 4. Crystals formed in about 1-3 minutes. The mixture is then slurried for 1 hour.

6) The crystals of the propylene glycol solvate of BL-S640 45 were filtered. Filtering is slow, with a

12.5-15.0 cm SS No. 604 paper is proposed. The crystals were then washed successively with 50 ml of 75% propylene glycol, 50 ml of methanol, 50 ml of acetone and then dried in vacuo at 56 ° C for 24 h. The biological yield was 80-95%.

Properties of the Propylene Glycol Solvate of BL-S640 Obtained:

55 a) Bio-test corresponded to 850-900 mcg / ml.

b) IR-NMR corresponded to a structure with 1.0-1.5 moles of propylene glycol (14-19% propylene glycol). No loss of the 3-triazole side chain could be demonstrated.

c)% water, K.F. = 1-3.0.

60 d) Crystal morphology = 100% crystalline (microcrystals, triangular shaped).

e) F = 182-184 ° C (D, hot step).

f) [a]: d5 (C = 1%, IN HCl) = + 53 ° C.

g) Water solubility corresponded to about 10 mg / ml in water

65 23 ° C.

h) Loss of bioactivity when stored at elevated temperatures: 10 ° C, 24h = <6%, 48h = <12%; 56 ° C, 1 month = <10%.

629 500 36

Preparation of crystalline methanol solvate from crystalline of the proposed structure and indicated that the product propylene glycol solvate contained no methanol but a trace of propylene glycol

50 g of the BL- propylene glycol solvate obtained above.

S640 were dissolved in 250 ml of a solution of 95 c / c vol / vol methanol

Slurried water at 22-25 ° C. Concentrated production of crystalline BL-S640 sesquihydrate and formation of HCl was then added to a pH of 1.1-1.5 other crystalline hydrates with rapid stirring, after which a solution or a Fast solution obtained 10.0 g of one Methanol solvate of BL-S640 (200 mesh) that was. The pH was adjusted to 1.7 with triethylamine and essentially free of propylene glycol, 7.5 g of activated carbon was added in 30-40 ml with slurry during 0.5 h of deionized water at ambient temperature, 20-25 ° C . The activated carbon was filtered and slurried with 75 ml of methanol 10 and an aqueous suspension was obtained from the. The wash solution was then added to the Fitrat. pH 3-4. After that, 40% NaOH was slowly brought to pH. The steps from the addition of HCl to this point should be within 6.3-6.7. The mixture was then finished at pH 5 h. The combined washing solution and the filtrate from 6.3-6.7 were stirred for 2 h. The crystals were filtered, stirred rapidly and washed with water within 5 min of triethyl and added at room temperature over a period of 24 h to a pH of 4.5. Crystallization 15 air dried and a 75-80% weight gain was obtained.

starts in about 1 -3 min. The mixture is then slurried for 1 h carrying 950-1000 mcg / mg of dihydrate crystals of BL and the crystals removed by filtration with S640. IR and NMR analyzes corresponded to the proposed 100 ml of methanol washed and in vacuo at 56 ° C during structure and indicated that the product did not dry any methanol for 24 h. The organic yield is 75-90%; the bioassay lasted but showed a trace of propylene glycol. H20,

corresponded to 850-900 mcg / mg; NMR-IR corresponded to 1 mol of Metha-20 K.F. = 6.56.

noi; % HiO, K.F. = 2-4.0. A sample of the crystalline dihydrate was at 37 ° C

- Air dried for 24 h and gave a crystalline sesquihydrate of BL-S640. H20 K.F. = 4.26.

Preparation of crystalline BL-S640 sesquihydrate A second sample of the dihydrate was kept at 45 ° C for 24

15 g of a methanol solvate of BL-S640 was air dried in 60 ml for 25 h and the crystalline sesquihydrate was obtained. Slurried water. The pH was adjusted to 6.5 by adding H2O, K.F. = 5.5.

4N NaOH was increased and the mixture was passed through a 200 A sample of the dihydrate was air dried at 56 ° C

Mesh sieve passed through. Thereafter, the reac- was slurried for 24 h and the crystalline monohydrate of ion mixture was slurried at room temperature for 2 h BL-S640. H20, K.F, = 4.38 (theory% water for monohy and the pH was maintained at 6.5 during this period. 30 drat = 3.75).

The crystals were filtered, washed with 20 ml of water and a sample of the dihydrate was air-dried in vacuo over P2Os at 37 ° C. for 24 h and pjan was dried at room temperature for 24 h to give the desired compound. The bioassay corresponded to 924 mcg / mg (in the crystalline hemihydrate of BL-S640. H20, K.F. = 2.63 (Theo-

% Water, K.F. = 5.26). NMR and IR corresponded to% water for hemihydrate = 1.91).

35

Claims (14)

629 500 PATENT CLAIMS 1. Process for the preparation of a new compound of formula I. h-n n 'C TLC in which A is hydrogen, hydroxyl, methyl or methoxy, R1 is hydrogen, sodium or potassium, R2 carboxyl or 2-furyl or an aliphatic, aromatic or heterocyclic radical on which a strongly acidic group, in the form of the free acid or as sodium or potassium salt is present, and E3 l, 2,3-triazol-5-yl, tetrazol-5-yl, 1,2,4-thiadia ch2-s-r- coor zol-5-yl, l, 3,4-thiadiazol-2-yl, l, 3,4-oxadiazol-2-yl or 1,2,4-triazoi-5-yl, each of these groups optionally including one or two lower alkyl groups is substituted, characterized in that one 1) a suspension in water or an inert organic solvent of an amphoteric cephalosporin of the formula II ch-c-wh coor chg-s-r3 ii 35 or a solvate thereof, in which A, R1 and R3 have the above definition, with an aldehyde of the formula R2-CHO, in which R2 was mentioned above, and a sodium or potassium base or triethylamine, sufficiently soluble in water or inert organic solvent, to adjust the pH Bring the value of the reaction mixture to between 5.5 and 8, treated and in solution the corresponding salt of the acid of formula 140 and 2) the sodium or potassium salt or the acid of the formula I is obtained from the solution. 2. The method according to claim 1, characterized in that the lower alkyl groups have 1-4 carbon atoms as substituents. 3. The method according to claim 1, characterized in that the amorphous starting cephalosporin of formula II is in the form of the methanol or propylene glycol solvate or the ses quihydrate. 4. The method according to claim 1, characterized in that sodium or potassium hydroxide is used as the base. 45 50 5. Procedure according to claim 1, characterized in that the compound of the formula I is lyophilized from the solution or precipitated from the solution by adding an organic solvent, preferably isopropanol, in which the salt is insoluble. 6. The method according to claim 1, characterized in that R2 for 2-furyl, 5-sulfo-2-furyl, 2-sulfophenyl, 4-methoxy-3-suIfophenyl, 4-hydroxy-3-sulfo-phenyl, 2-carboxy -methoxyphenyl, 4-carboxymethoxyphenyl, 3-hydroxy-4-carboxy-xyphenyl, 4- (2'-carboxy) vinylphenyl, carboxyl, 2-carboxyphenyl, 3-carboxyphenyl or sulfomethyl. 7. The method according to claim 6, characterized in that R2 is 2-furyl or 5-sulfo-2-furyl. 8. The method according to claim?, Characterized in that R3 stands for l, 2,3-triazol-5-yl. 9. The method according to claim 1 for the preparation of a compound of formula IV coor w i h IV 3rd 629 500 wherein R1 is sodium or potassium, characterized in that 13. The method according to claim 12, characterized in one draws that the solution obtained in stage 2) before 1) filtered a suspension of 7- [Du-amino-a- (p-hydroxyphenyl) - precipitation in stage 3), preferably before filtering with acetamido] -3- (l, 2,3-triazol-5-ylthiomethyl) - 3-cephem-4-carbon activated carbon treated. acid or a solvate or hydrate thereof in a suitable s 14. Process according to claim 1 for the preparation of an inert organic solvent, said compound having the formula IV given in claim 9, Solvent is a solvent for the triethylamine salt characterized in that a suspension of the corresponding Furfural reaction product of the aforementioned 3-C-ephem-4-car-hurrying starting 3-cephem-4-earbonic acid or a solvate acid and a non-solvent for the alkali metal salt or hydrate thereof in a suitable inert organic Formula IV is, io solvent used, said solvent being a 2) this suspension with furfural and sufficient triethylamine is nonsolvent for the alkali metal salt of the formula IV, which is treated, with formation in solution of the triethylamine salt of the suspension with furfural and a 3-cephem-4-carbon product called solvent-furfural reaction product Chen treated sodium or potassium base, wherein said base acid and is used in sufficient amount to the reaction mixture 3) Bring the alkali metal salt of the formula IV from the solution to the pH value between 5.5 and 8 by 15, and the product addition of a solvent-soluble sodium or potassium base of the formula IV is obtained from the reaction mixture. fails. 10. The method according to claim 9, characterized. 15. The method according to claim 14, characterized in that the starting 7- [Da-amino-a- (p-hydroxy- that the starting 3-cephem-4 -carboxylic acid in the form of phenyl) -acetamido] -3- (l, 2,3-triazol-5-ylthiomethyl) -3-cephem-2o of the methanol or propylene glycol solvate or sesquihydrate 4-carboxylic acid in the form of the methanol or propylene glycol - Is present, the inert organic solvent methanol and the solvate or sesquihydrate used. Base used in stage 1) sodium or potassium 2-ethylhexa- 11. The method according to claim 10, characterized thereby. characterized in that the solvent of the suspension methanol and 16. Method according to claim 14, characterized the sodium or potassium base in stage 3) sodium 2-ethylhexa- 25 indicates that one is a suspension of the methanol solvate of noat or potassium 2-ethylhexanoate. Starting 3-cephem-4-carboxylic acid used in methanol, the 12. The method according to claim 10, characterized in that the suspension treated with a molar excess of furfural is characterized in that the suspension in stage 2) is obtained with a molar and the product of the formula IV is filtered. Treated excess of furfural and the alkali metal salt of Formula IV from the solution by adding a solution of 30 17. The method according to claim 1, for the preparation Sodium or potassium 2-ethylhexanoate in isopropanol fails. a compound of the formula h < / ^ h c- hn / ii c4i Rc ch2-s • n-ch- COOR wherein R1 is hydrogen, sodium or potassium and R2 carboxyl radical, to which a strongly acidic group in the form of or 2-furyl or an aliphatic, aromatic or heteroey sodium or potassium salt is linked. 18. Ancestors according to patent claim I for the preparation of a document; of the formula r2 n- chp-s- <Tï * coor1 • ch ' ch- coor HNX X-H r2 hn ii- xc4i r2 6y é ' n- • c-ch- Y coor1 ch2-s- II n * / N- -N> -cho-s 2-ö ~ X or n i n-ch COORJ 629 500 where A water t. 1 lydroxv. Mollivi or Metlioxy. R 'hydrogen. Sodium or potassium uikI RJ CarboxyWnlor2-Furvl or an aliphatic, aromatiselier or heterocyclic residue ho c- 0 ÌI -c N- H2 ÏÏN .11 % c4i is to which a strongly acidic Ci group is bound in the form of the sodium or potassium salt. I'J. Verrahrcn according to claim 1 for the production of a Yerbindmm of the formula É2 0 </ -n < ch2-s- coor ,1 ch2 ~ s COOR ch- -n II N hît. .il r2 or coor1 hn, k- \ 4i RS 0 ^ S \ N- —Ch ^ -s- ç-ch- K c00rj wherein R 'is hydrogen. Is sodium or potassium and R-carboxyl or 2-furvl or an aliphatic, aromatiselier or heteroeyclic radical. to which a strongly acidic group is bound in the form of the sodium or potassium salt. 629 500 6 20. The method according to claim 1 for the production of a compound of I'orinel h 0 ho fVj ii hm m H2 N- -ÏÏ J Ì COOR hn m- xc4 E2 & coor1 h ho -O- 0 II ■ c 1 , N- N- CT sy ch2-s- ch < COOE1 where R is hydrogen. Sodium or potassium and Rr carboxyl sodium or potassium salt is bound. or 2-furyl or an aliphatic, aromatic or heteroev. 21. Process according to claim 1 for the preparation of a clic radical, in which a strongly acidic group in the form of the compound of formula a. or hn n- Xc4î f r2 0 ' & -N ch2-s- n / ch- COORJ 629 50 wherein A is hydrogen, hydroxy, methyl or methoxy, R1 is hydrogen, sodium or potassium and R "carboxyl or 2-furyl or an aliphatic, aromatic or heterocyclic radical to which a strongly acidic group is attached in the form of the sodium or potassium salt.