FI63030C - FOERFARANDE FOER FRAMSTAELLNING AV SOM ANTIBIOTER ANVAENDBARA SUBSTITUERADE N-METYLENDERIVAT AV TIENAMYCIN - Google Patents

Description

1 63030

Process for the preparation of substituted N-methylene derivatives of thienamycin useful as antibiotics

This invention relates to a process for the preparation of substituted N-methylene derivatives of thienamycin useful as antibiotics of the formula:

10 OH

- / N - SCH CH_N = C-R

I 2 2 I 1 2 I

I NR R

- --COOR '

15 O

wherein R is a hydrogen atom, a lower alkyl group, a phenyl, pyridyl, thiazolyl, trifluoromethyl, amino, azido, N-NH, alkyl or aminoalkyl group or a group -C 1-4 is a hydrogen atom, lower alkyl group, lower alkenyl group, lower alkylthioalkyl group, benzyl, dialkylamino, trifluoroethyl, carboxymethyl, alkoxy or nitro group;

2 12 25 and R is a hydrogen atom or a lower alkyl group or R and R

together with the nitrogen atom form a piperidyl ring and R1 is an alkylbenzyl, alkenyl or acyloxyalkyl group.

Thienamycin is described in U.S. Patent No. 30,399,357.

Thienamycin has the following structure:

OH

-SCH_CH_NH_ 35 I I 2 2 2 I II '

-N-U COOH

2 63030

Substituted N-methylenetienamycin derivatives can also be represented as an internal salt of the formula

OH O

5 I Cf) 1 H'— 1 2

- SCH2CH2N2C - ^ NR R

For the sake of simplicity, the esters prepared according to the invention are represented in the examples by the symbol:

QH

15 Th - N = C-X 1 '

Y

* - coor 'where "Th" denotes the bicyclic backbone of thienamycin and wherein the hydroxyl, amino and carboxyl groups of the compound are indicated.

New antibiotics are constantly needed. Unfortunately, there is no static efficacy of a particular antibiotic because the large-scale use of any such antibiotic selectively results in the emergence of resistant strains of pathogens. In addition, the disadvantage of the known antibiotics is that they are effective only against certain species of microorganisms. Thus, research to find new antibiotics 30 continues.

Surprisingly, it has been found that the compounds according to the invention are antibiotics with a broad spectrum of action which can be used in animal and human therapy and in non-living systems. The new antibiotics 35 are very broad-spectrum and are effective against both gram-positive bacteria (such as S. aureus, Strep. Pyogenes and 3 63030 B. subtilis) and gram-negative bacteria (such as E. coli, Proteus morganii, Klebsiella / Serratia and Pseudomonas).

The starting material compound is prepared by reacting a compound of formula

X

-ΓΤ-SCH-CH.NH.

I I 2 2 2 III

10 J-N-COOH

O

or a suitable O- and / or carboxyl derivative thereof for reacting with NR1R2 of formula

«O IV

With an imidoester according to RCX ° R "20, wherein X ° R" is a leaving group, wherein X ° is O or S and R "is lower alkyl 1 2 or the like and R, R and R are as defined above or R1R2N® »CX '£) I x' · v

R

With an imidohalide according to claim 30, wherein R, R1 and 2R are as defined above or in the formula X ° R "

35 r1r2n-c = n-r VI

With a compound of 4,63030, wherein X 0, R ", R, R 1 and 2 R are as defined above.

The overall synthesis of thienamycin is described in parent application No. 763308 (Patent No. 62,306).

Results of Pharmacological Experiments The lowest effective concentration of the new thienamycins and reference compounds was determined using standard gram-positive and gram-negative bacteria. The results are shown in the following tables.

5 63030 om vo • 'Τ n vo ro r- vo' d 'tOtOco ^^^^^ nJlOCQ O T- cm ro o CO 2 OCBPO Cr m Ό 3 m en in 3 ^ co vo ro r "m« n ( 1) tl) CM «· - - - -

10 CO K O r- CM ro O

Λ 2

VO

σι t- cm vo p VO CO CM CO "S '*“ 1) 03 CM »·« · »« * ·

P (0 ffl O O O O O

0 0 2 m co • Λ O

to o P Γ "3 -— · pop * - r ~ oo r- 3 P Cl) VO I— t— CM CM. r- C0 E 4-3 CM *“ »* -

P C CO O O O O O

> tn W 2 P ^

M P

(0 to vo co r- co cm en 3 is CM T- CM ro o C 3 cm ^ »s» v

φ 10 P «O O O O O

C -H rH 2

• H O O

P P O

H Q) -H · CM

<DCUWcor ~ co on ro OP p p ro p vo * - .¾, y (0 cm · * · * »^« · X cd> m o o o o o 3 X! (0 2

P P P

3 P P

«0 3 3

^ ^ P

Ή r * T- i "r * (N

C <0 CO O O O O O

<i)> tn cm - - - - - P 3 ra o o o o o CC Φ 2

P P P

P e 3 to φ (0 m

> 1 P OO ΓΟ P CM P CM

Era · ση o o o o o (0C / 1CM "» ****

C ra O O O O O

Φ 2

P

P

Φ

C P

Cm to cm ro • HP ra ra a a a ra to 3 u 3 Λ ra> 1 -

B

to H

φ φ '"ra ra a a a a p> u

P Φ P (0 M X

P - ro cm e _ a Y Λ a h ra a uija a 63030

C/O

3 a co -h

«H iH

O O r- m o 4J υ - «. - s *. i

• H · O O "Sf O. O

ή ° * M V

2 2> Ή “« e Τί +) \> c + j Cn = 3 a.

Ti os Λί> -

ti cc «H

·% 2 (0 oo «0 <N> CO *.

te a o- o m S U aajroino oor- OJ fsj -H p «- v | .. ..

5 W C 3 o o ** oo 71 U <D m rr

^ CO K -H

$ I CN * W O

s i <y Ä -H __ .¾ -p a e

rH flJ

2 β 'as - / O H

H A s tJ a os co ® tn

g · -H (0 CO CO CO (A

§ cp β C

2 C cd co cd co co o · a O M · Η Jm -h -h 5 «-p υ -p υ υ r |

-P

a 0) • o a ro m en m ro m

7, · sKSSKtCW

2 <D = UCJUUUCJ

g -P OS I I I I I

S W oooooo 2 -H uuuuuu t- · X! > 1 a a a P ro ro -P = a a a O O a

• H OS O O O CO CO., LTD

Λ5 O O

U '' a E-c 7 63030

Thus, the new compounds are at least as active as the known thienamycins.

As is known, thienamycin has an excellent antibacterial effect. Unfortunately, however, its chemical stability is poor and, in addition, it is inversely proportional to the concentration (Kropp, H. et al., "Thienamycin, A new β-lactam Antibiotic. II. In Vitro and In Vivo Evaluation", 16th Interscience Conference on Antimicrobial Agents and Chemotherapy, Chicago, 111, 1976). Amide-10 derivatives of thienamycin have 5-10 times better stability than thienamycin and equally good β-lactamase stability.

The new compounds are thus valuable antibiotics against various gram-positive and gram-negative bacteria and thus can be used as human and veterinary drugs. The compounds of the invention may be used as antibacterial drugs for the treatment of infections caused by gram-positive or gram-negative bacteria, e.g. Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Serratia, Salmonella typhosa, Pseudomonas and Bacterium Proteus. They can also be used as aqueous compositions in concentrations of 0.1 to 100 parts of antibiotic per million parts of solution for the purpose of inhibiting the growth of harmful bacteria in medical and dental equipment as well as as bactericides in industrial use, e.g., water-based paints and paper mill circulating water.

The compounds of the invention may be used alone or in combination as an active ingredient in a variety of pharmaceutical preparations. These antibiotics and their corresponding salts can be used as capsules or tablets, powders or liquid solutions or suspensions or elixirs. They can be administered orally, intravenously or intramuscularly.

The amount to be administered depends largely on the condition of the subject to be treated and the weight, dosage and frequency of the patient, with parenteral administration being preferred for conventional infections and oral administration for intestinal infections being preferred. Generally, a daily oral dose will comprise about 2 to 600 mg of active ingredient / kg of body weight to be administered in one or more doses per day. The preferred daily dose for an adult human is about 15-150 mg of active ingredient / kg of body weight.

The compounds of the invention may be administered in a variety of unit dosage forms, e.g., solid or liquid oral dosage forms. The unit dosage compositions, either liquid or solid, may contain from 0.1 to 99% of active ingredient, preferably from about 10 to 60%. The composition usually contains about 15 to 1,500 mg of active ingredient? however, it is generally preferred to use a dosage amount of about 100 to 1,000 mg. For parenteral administration, unit dosage forms will usually comprise a pure compound in a slightly acidified sterile aqueous solution or a soluble powder to be dissolved before use.

20 Example 1 Γ0Η NH Tr «

Th-NHC ^ BrCH 2 (O 2 -C (CH 3) 3 Th - NHC Preparation of benzyl ester N-Benzimidoyl-thienamycin (3.2 g) is suspended in hexamethylphosphoramide (75 μl) containing p-tert-butyl-benzyl bromide (3.8 μl) and stirred magnetically. At 22 [deg.] C. After 45 minutes, a solution is obtained which is stirred for a further hour, then the product is precipitated with ether solution and the crude product is chromatographed on a 9,630,302 [mu] m thick silica gel plate developed with chloroform / ethanol (7: 3). with an R f value of 0.6 is removed and eluted with ethanol to give p-tert-butylbenzyl ester hydrobromide of N-benzimidoyl-thienarazine Mass spectrometric analysis: m / e 521 (M +) 487, 444, 418 , 341, 323, 297, 226, 147.

Preparation of the starting material, N-benzimidoyl-thienamycin} 10 ™ p * H3C-O-C-0? 2 - = -> Th - N-C ^ 15

Thienamycin (59 mg, 212 μmol) is dissolved in a solution of 33% Ν, Ν-dimethylformamide and 0.05 (pH 7.0) in phosphate buffer (4.5 ml) and the pH is adjusted to 9.5 using 2.5 sodium hydroxide and an automatic dosing burette.

The solution is stirred magnetically at 25 ° C and methyl benzimidate hydrochloride (340 mg, 1981 μmol) is added in one portion. After 30 minutes, the solution is extracted twice with an equal volume of chloroform and the pH is adjusted to 7.0 with dilute phosphoric acid solution. The buffered solution is chromatographed using XAD-2 resin (65 mL). The column is eluted first with water and then with 10% aqueous tetrahydrofuran. This fraction is concentrated to half volume and lyophilized to give 50 mg of product. The electrophoretic mobility (50 V / cm, 30 20 min, pH 7 / 0.1 phosphate buffer) is 1.5 cm per anode. UV λmax = 300 nm (£ 6960), pH 7 / 0.1 phosphate buffer.

10 63030

Example 2 r-OH © 5 ^ m2

Th - NHCV-

Preparation of N-benzimidoyl-tlenamycin 3-methyl-2-buten-1-yl ester 15 N-Benzimidoyl-thienamycin (5.9 mg) is dissolved in hexamethylphosphoramide (100) containing 1-bromo- 3-methyl-2-butene (4.8, ul) and triethylamine (0.5 .mu.l) and stirred magnetically at 22 ° C. After 1 hour, the reaction product is chromatographed on a 250 μm thick silica gel plate developed with chloroform / ethanol (8: 2).

The rhinitis with an Rf value of 0.1 to 0.3 is removed and extracted with ethanol. After precipitation as a solid, benzimidoyl-thienamycin 3-methyl-2-buten-1-yl ester hydrobromide is separated from the ethanol / chloroform solution with hexane.

25 Example 3

r-OH o f-0H

+ ^ NH2 f + .NH2Br

Preparation of p-N-formimidoyl-thienamycin pivaloxymethyl ester hydrobromide Th - NHC BrCH, OCC (CH.,), Th - NHC '30 ~ - -> Thienamycin (10 mg) is dissolved in hexamethylphosphoramide (200 μl) containing bromomethyl pivalate (10 μl) and triethylamine (1 μl) and -! f f 11 63030 is magnetically stirred at 22 ° C. After 2 hours, the hexamethylphosphoramide solution is dissolved in 2 ml of methylene chloride and the product is precipitated with hexane / ether solution (1: 1). The precipitate is dissolved in a 10% aqueous solution of tetrahydrofuran and chromatographed on an XAD-2 resin column. The pivaloxymethyl ester of N-formimidoyl-thienamycin is separated in solid form by eluting the column with tetrahydrofuran and lyophilizing.

Preparation of the starting material, N-formimidoyl-thienamycin 10

OH

—-SCH.CH-N 2 C-NH-I 2 2 | 2

1 I H

^ -N - COOH

15

Thienamycin (517 mg) is dissolved in 0.1 N phosphate buffer (pH 7, 25 mL) and cooled in an ice bath with magnetic stirring. The solution is adjusted to pH 8.5 using 2.5 N sodium hydroxide solution (automatic 20 burettes). While maintaining the pH at 8.5, methyl formimidate hydrochloride (711 mg) is added portionwise over 2-3 minutes. After 10 minutes, adjust the pH of the solution to 7.0 using 2.5 N hydrochloric acid. The solution is chromatographed on an XAD-2 resin column (150 ml) eluting with water. The N-formimidoyl-thienamycin derivative is initialized in 1.5 to 2.0 column volumes (200-300 ml) and lyophilized to a white solid (217 mg). UV (pH 7, 0.1 N phosphate buffer) λmax = 297 nm (8590). IR (Nujol) 1767 cm -1 (β-lactam). NMR (D 2 O) δ 1.37 (d, 30 J = 6 Hz, CH 3 -CH), 3.0-3.75 (m, -CH 2 -), 4.2 - 4.8 (m,

NH

C ^ jj, Cgjj, C ^ jj), 7.86 (s, -C-H).

i

Claims (2)

i2 63030 Claim: A process for the preparation of substituted thienamycin-substituted N-methylene derivatives of the formula OH - / N.-SCH-CH N = C-R 10 [| NR 1 R 2 -N-COOR 10 wherein R is a hydrogen atom, a lower alkyl group, a phenyl, pyridyl, thiazolyl, trifluoromethyl, amino, azido-NH 1 alkyl or aminoalkyl group or a group C, R is OCH3 is a hydrogen atom, a lower alkyl group, a lower alkenyl group, a lower alkylthioalkyl group, a benzyl, dialkylamino, trifluoroethyl, carboxymethyl, alkoxy or nitro group, and 2. R is a hydrogen atom or a lower alkyl group, or R and 2 R together with the nitrogen atom form a piperidyl ring and R 'is an alkylbenzyl, alkenyl or acyloxyalkyl group, characterized in that the compound of formula OH 30 -SCH CH-NCR il kV 11 I-NL-COOH CY is esterified 35 2 2 where R, R and R have the same meaning as above.