DE2815157A1 - 3- (2-AMINOAETHYLTHIO) -6-AETHYL-7-OXO-1- AZABICYCLO SQUARE CLAMP ON 3.2.0 SQUARE BRACKET TO HEPT-2-EN-2-CARBONIC ACID - Google Patents

Description

, .- ING. WALTSK ABITZ,. DIETER F. MORF PL.-PHYS. M. GRITSCHNEDER

ntairwälte

April 7, 1978

Postal address Postfach 8ΘΟ1Ο9, 80OO Munich 86

Pienzenauerstraße 28 Telephone 98 32 22

Telegrams: Chemindus Munich Telex: COJ 523992

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MERCK & CO., INC. Rahway, New Jersey, V.St.A.

3- (2-Aminoethylth.io) -6-ethyl-7-oxo-1-azabicyclo / 3.2.Q7! HLept-2-en-2-carboxylic acid

809842/0898

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"■ -. S- ' " ..28T5157

The invention relates to 3- (2-aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo / 3.2.0 / hept-2-en-2-carboxylic acid with the formula I.

6 '

SCH ₂ CH ₂ NH ₂ .: - ";. COOH ί

and their pharmacologically acceptable salts, which compounds are useful as antibiotics. The invention also relates to Methods for the preparation of the compounds (I) and Medicines containing the compounds mentioned.

"3- (2-Aminoethylthio) -e-ethyl-T-oxo-i-azabicyclo / 3.2.O / hept-2-en-2-carboxylic acid (I) "is expediently derived from the antibiotic thienamycin with the formula II

SCH ₂ CH ₂ NH ₂ j (H)

COOH "!

produced by splitting off the 8-hydroxyl group.

Thienamycin is described in U.S. Patent 3,950,357. To this Patent is expressly incorporated herein by reference as thienamycin as a starting compound for the synthesis of the invention can serve according to connections.

Thienamycin and all of its isomers (in pure form or as Mixtures) can also be obtained by the US patent application Ser. 833,210 (September 15, 1977) (corresponds to German patent application P 27 51 597.5) described total synthesis getting produced. Reference is made to this registration here insofar as it connects all of the output

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fertilize to produce the compounds according to the invention Serving isomers of (I) and their preparation describes.

The invention also relates to those for making the compounds (I) suitable intermediates with the general formulas III and IV

in which R and R 'each represent a hydrogen atom or a Easily split off protecting groups mean (R and R * are defined in more detail below).

There is a constant need for new antibiotics. There is namely, no static effectiveness of certain antibiotics, because a constant and extensive use of any one "Antibiotic selectively leads to the appearance of resistant strains of pathogenic microorganisms. The known antibiotics also have the disadvantage that they are only effective against certain microorganisms. It will therefore constantly researching new antibiotics.

The invention is therefore based on the object of a new antibiotic and its pharmacologically acceptable salts to make available which compounds are suitable for human and veterinary medicine as well as for the treatment of inanimate Systems. Such antibiotics have activity against a variety of pathogenic microorganisms, to which both gram-positive bacteria (such as S. aureus, Strep, pyogenes or B. subtilis) as well as gram-negative bacteria (such as E.coli, Proteus morganii, Serratia or Klebsiella). Furthermore, the invention is intended to provide methods for the production of the antibiotics mentioned and their non-toxic, pharmaceutical

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macologically compatible salts as well as these antibiotics containing drugs are created.

The compounds of the invention are generally made according to prepared according to the following reaction scheme:

CH ₃ CH;

base

(Hb)

reduction

N IL-COOH

(HI)

COOH

The rest of the OR of the connection (lib) is easily done with the help of a The base is split off, the enlactam (III) being obtained. This is converted into the compound according to the invention by reduction (I) received. The OR radical is typically a sulfate or sulfonate group.

In a more specific respect, the 3- (2-aminoethylthio) -e-ethyl-T-oxo-i-azabicyclo / 3.2.07hept-2-en-2-carboxylic acid according to the invention is used (I) suitably prepared according to the following reaction scheme:

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OH

-N LL COOR

·? ■

Sulfonation

(Ila)

COOR

(lib)

ase

CH ₃ CH

J N LL COOR

⁰ (III)

H ₂ / Cat _v

S chut ζ groups split off

COOR

(IV)

COOH

(D

In the above reaction scheme, R "means CHlV ¹ H (where M is a
Hydrogen atom, or any harmless cation,
such as an alkali metal ion or an organic base or an ion derived therefrom) or an alkyl or aryl radical (such as a methyl or toluyl group), R an easily removable blocking or protective group for the carboxyl group and R 'an easily removable N- Blocking or protection group. Such protected or blocked thienamycins (Ila) are in the
USA patent application serial no. 733.65p (filed on October 18, 1976). Reference is made to this application here with regard to the production of (ila). Preferred carboxyl protecting groups R are, for example, the benzyl group and ring-substituted benzyl groups of the general formula

-X

in the X is.B. a nitro group or a lower alkoxy radical, such as is a methoxy group. The benzyl group is used as a protecting group particularly preferred. Specific examples for suitable +) = German patent application P 27 24 56Ο.9

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nete N-protective groups R 'are the bromo-tert.-butoxycarbonyl-, Chloro-tert-butoxycarbonyl-, bromoethoxycarbonyl-, benzyloxycarbonyl- and p-nitrobenzyloxycarbonyl group. One special preferred Ν protecting group is the bromo-tert.-butoxycarbonyl- group.

As illustrated by the above reaction scheme, a suitably protected thienamycin (Ha) is converted into the corresponding sulfonate or sulfate ester (lib) by methods described in the aforementioned US patent application Ser. 733,655 (filed October 18, 1976), which are incorporated herein by reference. Typically, the intermediate (lib) is prepared as sulfonate by reacting (Ila) with a sulfonating agent (such as methanesulfonyl chloride or toluenesulfonyl chloride) in a solvent (such as methylene dichloride, tetrahydrofuran, dioxane or chloroform) in the presence of a base (such as triethylamine, pyridine, 4-dimethylaminopyridine or sodium hydride) for 1 to 10 hours at temperatures of -15 "prepared 25 ⁰ C. Sulfatestervarianten of intermediate (lib) (R" = to -0®M®) are also suitable as starting compounds and have the general formula

COOR
Ö '

in which M is a hydrogen or alkali metal ion or an organic base or an ion derived therefrom and R ^f each Weil have the meaning given above. Such sulfate or sulfuric acid esters are also described in the aforementioned US patent application Serial No. 733,655.

As far as the reaction of the intermediate (lib) is concerned, the radicals R and R ^{f can} also represent hydrogen atoms in addition to the aforementioned protective groups. Such intermediates

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(lib) are protected by splitting off the protective groups from the Form of (lib) prepared by conventional methods described below.

The sulfate ester variants of the intermediate products (lib) (R "= OH) are also available as natural products. Such products are in the US patent applications Serial Kos. 767,723 and 767,920 (each filed on February 11, 1977) (which the German Patent applications P 28 05 701.1 and P 28 05 724.1). The above-mentioned registrations are insofar here Reference is made to the preparation and isolation of the sulfate ester intermediates which can be used in the process according to the invention (lib) describe.

As can be seen from the reaction scheme above, the enlactam intermediate (III) is obtained from (Hb) by reaction with a base (such as with NaHGO · ,, EpHPO *, triethylamine or 1,5-diazabicyclo / B ^ .Q / undec-5 -en) in a solvent (such as methanol, tetrahydrofuran or dioxane) for 1 to 20 hours at temperatures of -20 to 25 ° C.

The 6-ethyl intermediate product (IV) is produced from (III) by reduction. (III) is preferably in a solvent (such as ethanol, ethyl acetate, dioxane or benzene) in the presence of a hydrogen catalyst (such as PtO ₂ , RuO ₂ or Pt / C) for 30 minutes to 5 hours at temperatures from 0 to 25 ° C and a hydrogen pressure of 1 to 4 atm. (1.01 to 4.05 bar). The intermediate (IV) can be converted into (i) by splitting off protective groups in the course of the hydrogenation described above; Depending on the nature of the protective groups R * and R, the splitting off of the protective groups can also be carried out by separate methods, such as by hydrolysis or hydrogenation, under different conditions. It should be noted that the

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Removal of protective groups according to the reaction scheme can also be carried out earlier on the intermediate (lib). The splitting off of protective groups can also take place as a one-step reaction in the conversion of (III) into (IV). Separate deprotection (IV-J · I) can be carried out by any of a number of known methods, such as hydrolysis or hydrogenation. The carboxyl-protecting group R is preferably by hydrogenation in a solvent (such as a lower-alkanol such as ethanol) in the presence of a hydrogenation catalyst (such as palladium, platinum or oxides thereof) for 1 to 10 hours at 0 to 5O ⁰ C and a hydrogen pressure from 1 to 40 atm. (1.01 to 40.5 bar) split off. The N-Schutζgruppe R * can also be split off by hydrogenation; when using the preferred bromo-tert-butoxycarbonyl group, however, deblocking is expediently achieved by heating a solution of (IV) or the compound (IV) freed from the carboxyl protective group in a solvent (such as ethanol, isopropanol or water) for 10 minutes up to 3 hours at 25 to 80 ⁰ C.

The compounds (I) according to the invention form with inorganic compounds and organic bases a variety of pharmacological compatible salts. Examples are metal salts, which are derived from alkali or alkaline earth metal hydroxides, carbonates or! -! bicarbonates derive, and Salzey which differ from primary, secondary or tertiary amines, e.g. monoalkyl- ■ amines, dialkylamines, triallcylamines, lower alkanolamines » Di-lower ~ alkanolamines, lower alkylene diamines, NjIT-diaralkyl-lower alkylene diamines, Aralkylamines, amino-substituted Hieder-alkanolens N ^ -di-lower-alkyiaminosubstitaiierten ITlower alkanols, amino, polyamino or guanidino-substituted Hxeder-al] ran {carbon) acids or nitrogen-containing heterocyclic amines.

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Typical examples are salts which, τοη! Sodium hydroxide, Sodium carbonate, i-sodium bicarbonate, potassium carbonate, potassium hydroxide? Calcium carbonate trimethylamine triethylamine, Piperidine, morpholine, quinine, lysine, protamine, arginine, Procaine, ethanol arain, morphine? Benzylamine, ethylenediamine, HjF-dibenzylethylenediamine, diethanolamine, piperazine damsel thylaminoethanol, 2-amino-2-methyl-1-propanol, theophylline or N-methylglucamine. Primary amino group salts of (i) with pharmacologically acceptable organic and inorganic acids are also included. Specific Examples of such salts are methanesulfonate, 2-naphthalene sulfonate, pamoate, 3-phenylpropionate, trimethyl acetate, tert-butyl acetate, p-toluenesulfonate, maleate, lactate, Cyclamate, pumarat, tartrate, oxalate, benzoate, acetate, succinate, citrate, glutamate, hydrochloride, hydrobromide, sulfate, Phosphate, n-acetylglycinate, benzenesulfonate, hexanoate, p-chlorobenzenesulfonate, Cyclopentane propionate, 1,2-ethane disulfonate, Gluroheptanoate, ethanesulfonate, o- (4-hydroxybenzoyl) benzoate and 2-hydroxyethanesulfonate; the salts can be used according to conventional Methods are made.

The salts can be monosalts, such as the reaction of 1 equivalent of sodium hydroxide with 1 equivalent of the product (I) be obtained lionosodium salt * or mixed disalts » The latter can be achieved by reacting 1 equivalent of a base with a divalent cation (e.g. calcium hydroxide) 1 equivalent of the product (I) can be generated. The inventive Salts are pharmacologically acceptable, non-toxic derivatives of the compounds (i), which as • active ingredients in suitable single dosage forms of drugs can be used. One can combine these salts with other active ingredients to make medicines with a broad spectrum of action.

ORIGINAL JNSPECTED

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The compounds "" of the invention (I and salts) are valuable antimicrobial substances that are resistant to various graih-positive and gram-negative pathogenic microorganisms are effective. For example, the free acid and, in particular, its salts, such as the amine or metal salts (especially the alkali metal or alkaline earth metal salts), valuable bactericides and can be used to eliminate susceptible

pathogenic microorganisms from medical and dental equipment, for the separation of microorganisms and for therapeutic purposes in human and veterinary medicine. I ^{1 For} the latter purpose, pharmacologically acceptable salts with inorganic and organic bases, such as are customary for the administration of penicillins and cephalosporins, can be used. Salts, such as alkali metal and alkaline earth metal salts or salts

of primary, secondary and / or tertiary amines are e.g. suitable for this purpose. These salts can be mixed with pharsa ecologically compatible liquid and solid diluents or carriers for suitable single dosage forms » like pills, tablets, capsules? Suppositories, syrups or. Elixirs, the production of which is carried out in the usual manner can be combined. . · '■

The new connections are - as mentioned - valuable, opposite various gram-positive and gram-negative bacteria effective antibiotics, which accordingly. in the Find human and veterinary medicine use. One can the compounds according to the invention are therefore considered to be antibacterial Use active ingredients to treat infections caused by gram-positive or gram-negative bacteria. ■ will call, e.g. against Staphylococcus aureus, Escherichia coli-, .Klebsiella_pneumoniae, Bacillus subtilis, Salmonella typhosa, Pseudomonas and Bacterium proteus. the antibacterial compounds according to the invention can also

■■ "'■ / ■ - ■; ■: .. ■:; ■■. ■ .-. -G -

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Not to be used as animal feed additives, food preservatives and disinfectants »Man. can the compounds e.g. in the form of -aqueous preparations with Concentrations of Q, 1 to 100 parts of antibiotic per Million parts! Solution to destroy or inhibit the Growth of harmful bacteria on medical and dental equipment or as technical or industrial bactericides, e.g. in aqueous paints or in wastewater, of paper mills to inhibit the growth of harmful ' insert bacteria.

The compounds according to the invention can be used in a wide variety of medicinal products as single active ingredients or can be used in combination with other active ingredients. The antibiotics (I) and the corresponding salts can e.g.

in porra of capsules? Tablets, powders or powder preparations, such as solutions, suspensions or elixirs can be used. Administration can be oral, "intravenous or intramuscularly »

The medicinal preparations preferably have one for Absorption in the gastrointestinal tract in a suitable form. Orally administrable tablets and capsules can be in unit dosage form exist and conventional auxiliary or Packaging agents, such as binding agents, e.g. syrup, gum arabic, gelatin, sorbitol, tragacanth or polyvinylpyrrolidone, ! Fillers, such as! Lactose, sugar (cane sugar), Corn starch, calcium phosphate, sorbitol or glycine, lubricants such as magnesium stearate, talc, polyethylene glycol or silicon dioxide, disintegrants such as potato starch, or compatible wetting agents, such as sodium lauryl sulfate, contain. The tablets can also be coated using conventional methods. The oral! Liquid preparations can be in enormous amounts of watery or oily suspen-

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sions, solutions, emulsions, syrups or elixirs exist or are offered as dry products »which are reconstituted or diluted with water or other suitable media before use. Such liquid preparations can contain conventional additives - such as suspending agents, eg sorbitol, syrup, methyl cellulose, glucose / sugar syrup * gelatine, hydroxyethyl cellulose? Carboxymethylcellulose, Aluminiunistearatgel or hydrogenated edible oils, for example almond oil, fractionated coconut oil, oily esters _9, propylene glycol or ethanol, or preservatives ,. For example p-Eydro ^ ybenzoic acid methyl ester or propyl ester or sorbic acid contain. Suppositories contain conventional suppository bases such as cocoa butter or another glyceride. . ..

Injection preparations can be in single dosage form in ampoules or in multi-dose containers with added preservatives. These preparations can as suspensions ^ solutions or emulsions in oily or aqueous media are present and packaging agents, like suspending agents, stabilizers and / or dispersing agents » contain. Optionally, the active ingredient can be in lOrm of a powder, which before use with a suitable medium, e.g. sterile, pyrogen-free water will. ■. ·,

The medicinal preparations can also be prepared in a Porm 'j

that are responsible for absorption through the mucous membranes:

the nose and throat or the bronchial tissues; .]

you can then expediently use the ΙΌπα of powdery J-

or liquid sprays or. Inhalants, lozenges -. j

(lozenges) or neck tinctures, etc.; eye and ''

Ear preparations can be used as individual capsules or in liquid form ^:

lighter or semi-solid or in the form of drops!

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. ° ^ AL _INSP 809842/0898 - ^t0

■ be turned. Topical (local) preparations can be found under Use of hydrophobic or hydrophilic bases formulated as ointments, creams, lotions, tinctures, powders, etc. will.

The medicaments according to the invention can be taken from a carrier other ingredients such as stabilizers, binders, antioxidants, preservatives, lubricants, Suspending agents, viscosity regulators or flavoring or odor correcting agents and coloring agents (flavoring agents) contain. Furthermore, the drugs can contain other active ingredients to achieve a broader spectrum of antibiotic activity contain. '; - "-" -'.

Veterinary medicine can use the drugs e.g. as intraniammary preparations using τοπ bases that are for provide a long-term effect or a rapid release, formulated will.

The dose to be administered depends to a large extent on the condition and weight of the patient or animal to be treated as well as the route and frequency of administration. The parenteral route is preferred for general infections, while the oral method is preferred for intestinal infections. will. An oral daily dose generally consists of about 15 to about 600 mg of active ingredient / kg of body weight for one or several application (s) per day. Daily doses become pure adults from about 80 to 120 mg active ingredient / kg body weight is preferred. - - -

The agents according to the invention can be administered in several individual doses sen, e.g. as solid or liquid, orally tolerated preparations, the preparations can be

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rate - (liquid or solid) 0.1 to 99 ? ° (preferably about:

10 Ms 60 ψ) contain active ingredient. The preparations contain ■ \

generally about 15 to about 1500 mg of active ingredient; preferred ;

however, doses will range from about 250 to 1000 mg. ·; In the case of parenteral administration, there is a single dose

form usually in a solution of the pure compound. · ■ '

in weakly acidified »sterile water or in a for ι

the! solution provided, soluble powder. !

The examples below are intended to illustrate the compounds that Explain the methods and the medicament of the invention in more detail, without any intention of limitation.

- Example 1

Production of IT — bromine tert. -butvloxy carbonyl-thi enamycin sodium as

Method A:

A solution of 190 mg of thienamycin in 15 ml of 0.1 M phosphate buffer and 15 ml of dioxane is kept at ⁰ ° C. Man. provides the solution with -In IT sodium hydroxide solution. to a pH of 8.5 to. 9 »0 and keeps it in this pH range, while it is - ■ added within 5 minutes with 480 mg of chloroformic acid bromo-tert-butyl ester. The mixture is 30 Hin. stirred, then neutralized with 1N HCl, (up to pH 7.0) and extracted with ether. The aqueous layer is separated off, evaporated to 10 ml and chromatographed on a Dowex 50x8 (urodium form) containing column (3.81 χ 25.4 cm or ϊ.5 χ Ϊ0 in.). The column is eluted with water, T13 mg of the desired product being obtained after freeze-drying.

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Method B:. - '. I.

A solution of 95 mg thienamycin in 10 ml 0.1 M phosphate buffer and 10 ml Diosan is kept at ⁰ ° C. One adjusts ^: the solution to a pH value of 8.5 to 9.0 and maintains it. ^; keep them in this pH range while getting 240 mg. Chloroformic acid bromobutyl ester added. The mixture is stirred for 30 minutes and then acidified with phosphoric acid (to pH 2.0). The acidified solution is extracted twice with 25 ml of ethyl acetate each time. The organic layer is separated off and back-extracted with 10 ml of aqueous sodium bicarbonate solution containing 30 mg of NaHCO ^. The aqueous layer, after freeze-drying, gives 30 mg of the desired product; NMR (60 MHz, DGO): $ ^1, 26 (d), 1.60 (s) 2.65 to 3.50 (m), 3.70 (s), and 3.90 to 4.20 (m); UY λ ° | _χ 303

example

Production of H-bromo-tert-butyloxycarbonyl-thienamycin-p-nitrobenzyl ester j

The lyophilized N-bromo-tert-butyloxyearbonyl-thieno, amycin-Natriumsals (100 mg) for 1 hour '. At 26 ⁰ C with 300 p-Uitrobenzylbromid day in 2 ml of hexamethylphosphoramide. touched. . The mixture is then diluted with 10 inl ethyl acetate and washed thoroughly with water. The organic layer is separated, dried over sodium sulfate and chroniatographed on two 250 μ-Sj.eselgel-GI 'thin-layer chromatography plates using ethyl acetate as the solvent (R ^ = 0.45), 50 mg of the desired product being obtained; IR (CDCl ₃ ): 1777 (β-lactam) and 1711 cm " ¹ (ester); UV λ ^ J ^{82101 2} ? ° ² ^" ¹ ^ ^{522 2211} J ^miR ( ⁰ BCl ₃ , 6Q IiHz): cT 1.38 (d), 1.58 (s), 2.60 to 3.80 (m), 3.78 (s), 3.90. "to 4.20 (yes), 5.30 (s), 7.55 (d) and 8.30 ppm (d).

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The benzyl ester is according to Example 2, but using the equivalent amount of benzyl bromide instead of p-nitrobenzyl bromide.

example

Preparation of N-benzyloxycarbonyl-thienamycin-benzyl ester

NHC-O-CH ₂ C ₆ H ₅

CO ₂ CH ₂ C ₆ H ₅

Dissolve 214 mg (0.79 mmol) of thienamycin in 22 ml of ice-cold deionized water, stir the solution while cooling with an ice bath, add 995 mg (11.85 mmol) NaHCO3 at once and stir the Approach for another 5 minutes. 17.6 ml of dioxane are then added and the solution is stirred for a further 5 minutes. Then add within 16 min. an ice-cold solution of 242 ing (1.42 mmol) Benzyl chloroformate in 5 ml of tetrahydrofuran in the form of five equal portions. The mixture is stirred for a further 15 min. in the cold, the reaction mixture is covered with 40 ml of ice-cold diethyl ether and acidified with cold 5% aqueous Η, ΡΟ ^ up to pH 6.8. You separate the layers from each other and extracted the aqueous portion with further cold diethyl ether (2 χ 30 ml). One layered on top the aqueous phase in 35 ml of ice-cold ethyl acetate and the mixture is stirred rapidly while cooling with an ice bath, the pH Set value to 2.4 with cold 10biger aqueous Η, ΡΟ ^. Separate the layers and extract the aqueous portion with more cold ethyl acetate (15 ml). The combined ethyl acetate solution is then washed with 20 ml

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ice cold sodium chloride solution, dry it in the cold over sodium sulfate and filtered. The piltrate contains N-benzyloxycarbonyl-thienamycin.

The ethyl acetate solution is mixed with ice-cold ethereal phenyldiazomethane (15 ml of a 0.25 M solution) and leave the resulting solution in the refrigerator for 1 hour. After evaporation of the solvent in vacuo, the residue becomes diluted twice with ethyl acetate to separate off the dioxane and exposed to a vacuum. Then you dissolve the residue in 5 ml of ethyl acetate and slowly dilute the solution with 5 ml of diethyl ether. The mixture is then left in the Stand in the refrigerator and then filtered off the precipitate formed. The filter residue is washed with cold diethyl ether and dry it in a vacuum. 16O mg of N-benzyloxycarbonyl-thienamycin-benzyl ester are obtained in the form a 3 gray-white solid.

The mother liquors and wash water are combined and evaporated in vacuo. The residue is rubbed with hexane (3 × 50 ml), dissolve it in ethyl acetate and pass the solution through a sintered glass funnel containing 2 g of silica gel to run. The silica gel layer is washed out with more ethyl acetate. The ethyl acetate solutions that have flowed through are combined, concentrated in vacuo to about 1 ml, diluted to 5 ml with diethyl ether and 2 days in the refrigerator ditched. The precipitate formed is then filtered off, washed with cold diethyl ether and dry it in a vacuum. This gives further N-benzyloxycarbonyl-thienamycin-benzyl ester (48 mg) in the form of white flakes.

The first product yield has the following characteristics: melting point 125 to 126 ^° C. (heatable microscope);

IR (CH ₂ Cl ₂ ) 3608, 3424, 1780, 1720, 1514, 1332, 1206 and 1137 cm ""¹;

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UV (dioxane) 317 (S 11,800) nm;

NMR (CDCl ₃ CT 1.34 (d, 3, CH ₃ ), 2.70-3.55 (m, 7, CH ₂ , SCH ₂ , NCH ₂ , H ₆ ), 4.16 (m, 2, H5, H8), 5.10 (s, 2, NCO ₂ CH ₂ ^), 5.29 (ABq, 2, CO ₂ CH ₂ ^) and 7.32 (m, 10, ArH) (0 = phenyl) .

Example 4 N-Benzyloxycarbonyl-thienamycin-benzyl ester

798 mg (2.93 mmol) of thienamycin are dissolved in 200 ml of ice-cold water and the resulting solution is stirred while cooling with an ice bath. Then 2.462 g (29.3 mmol) of solid NaHCO ₃ and, after a few minutes, 200 ml of dioxane are added all at once. The mixture obtained is stirred in the cold for 3 minutes and then treated dropwise over 10 minutes with a solution of 750 mg (4.4 mmol) of benzyl chloroformate in 12 ml of anhydrous dioxane. The mixture is then stirred for a further 10 minutes in the cold and the reaction mixture is then extracted twice with 120 ml of ice-cold diethyl ether each time. The aqueous portion is covered with 100 ml of cold ethyl acetate and stirred vigorously, acidifying with cold 1 m H ₂ SO ^ (up to pH 2.3). The layers are then separated from one another and the aqueous phase is extracted with further, cold ethyl acetate (2 × 30 ml). The combined ethyl acetate solutions are washed with 50 ml of ice-cold sodium chloride solution and then extracted with 58.6 ml of ice-cold 0.05N LiOH. The aqueous LiOH extract is evaporated on a rotary evaporator in vacuo to separate off the ethyl acetate and then freeze-dried. This gives 1 g of crude N-benzyloxycarbonyl-thienamycin-idthium salt in the form of a pale yellow solid.

A sample (864 mg) of the aforementioned lithium carboxylate is suspended in 15.7 ml of anhydrous hexamethylphosphoramide and the suspension is mixed with 1.15 ml of benzyl bromide.

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■ Sa · 28 151 S?

The mixture obtained is stirred for 2 hours at 25 ^° C. under nitrogen. The mixture is then diluted with 350 ml of ethyl acetate and washed with water (2 × 300 ml), 5% sodium bicarbonate solution (150 ml), water (2 × 150 ml) and sodium chloride solution (150 ml). The ethyl acetate solution is then dried over magnesium sulfate and filtered. The filtrate is evaporated in vacuo to a gummy residue. To separate off the excess benzyl bromide, this is rubbed with hexane (5 × 10 ml) and then dried in vacuo, 817 mg of a pale yellow solid being obtained. This crude product is dissolved in 15 ml of warm ethyl acetate. The solution is diluted with 15 ml of diethyl ether, inoculated and left to stand in the freezer overnight. The precipitate formed is then isolated, washed with cold diethyl ether and vacuum-dried. This gives 405 mg of N-benzyloxycarbonyl-thienamycin-benzyl ester in the form of a gray-white solid.

The mother liquors and wash water are evaporated in vacuo to an oily residue which is warmed in about 5 ml Ethyl acetate dissolves. The solution is diluted with about 10 ml of diethyl ether and then in the freezer for several days left to stand, wobej. one a second product yield (106 mg) was obtained as a yellow solid.

example N-Benzyloxycarbonyl-O-methanesulfonyl-thienamycin-benzyl ester

OMs

I_JL

Jl -U-CO ₂ CH ₂ C ₆ H ₅

Dissolve 50 mg (0.1 mmol) of N-benzyloxycarbonyl-thienamycin-

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.23. 2,5157

benzyl ester in 3 ml of anhydrous CHpCl ₂ and stir the solution while cooling with an ice bath in a nitrogen atmosphere. 27.9 μΐ (0.2 mmol) of triethylamine are added to the solution and a solution of 17.2 mg (0.15 mmol) of methanesulfonyl chloride in 0.4 ml of Meüiyüßndichlorid is then added dropwise over the course of 5 minutes. The resulting solution is stirred in the cold for 90 minutes, _{diluted with 10 ml of ice-cold CH 2} Cl ₂ , washed with 5 ml of cold water, 2 ml of cold 0.1 M pH 3 phosphate buffer and 5 ml of cold 2 S sodium bicarbonate solution, dried over magnesium sulfate and filtered. The solvent is evaporated from the filtrate in vacuo, 57 mg of N-benzyloxycarbonyl-O-methanesulfonylthienamycin benzyl ester being obtained in the form of a pale yellow oil with the following characteristics:

NMR (CDCl ₃ ) 1.56 (d, 3, CHCH ₃ ), 2.82-3.52 (m, 7, CH ₂ , SCH ₂ , NCH ₂ , H6), 3.00 (s, 3, CH ₃ SO ₂ ), 4.28 (d of t, 1, H5), 5.08 (d of q, 1, H8), 5.09 (s, 2, NCO ₂ CHjZi), 5.26 (ABq, 2, CO ₂ CH ₂ JZi) and 7.32 (m, 10, ArH).

In an analogous manner, N-bromo-tert-butyloxycarbonyl-O-methanesulfonyl-thienamycin-p-nitrobenzyl ester is obtained, if instead of the N-benzyloxycarbonyl-thienamycin benzyl ester the equivalent amount of N-bromo-tert-butyloxycarbonylthienamycin-p-nitrobenzyl ester begins.

example

Benzyl 6-ethylidene-3- (2-benzyloxycarbonylaminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylate

CH ₃ CH

A solution of 44 mg (0.077 mmol) of N-benzyloxy-

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carbonyl-O-methanesulfonyl-thienamycin benzyl ester in anhydrous methanol with 12.9 mg (0.154 mmol) of powdered sodium bicarbonate and the resulting mixture is stirred for 100 min. at 25 ⁰ C. Thereafter, the mixture is diluted with 15 ml CH ₂ Cl _2, three times washed with 5 ml of water each time, dried over magnesium sulfate and filtered. The filtrate is evaporated in vacuo to an oil (33 mg). The crude oil is purified by thin layer chromatography on a 250 μm × 20 cm × 20 cm silica gel GF plate using ethyl acetate / chloroform (3: 1) as the developing agent. The major UV visible band is separated and eluted with ethyl acetate to give an oil (24 mg). This material is again chromatographed in the manner described; 16 mg of benzyl 6-ethylidene-3- (2-benzyloxycarbonylaminoethyl) -thio-7-oxo-i-azabicyclo [3.2.0] hept-2-ene-2-carboxylate are obtained in the form of a light yellow oil with the following characteristics :

IR (CHCl ₃ ) 3610, 1779, 1715, 1508, 1330, 1271, 1178 and 1121 cm-1; .

NMR (CDCl,) cf 1.80 (d, 3, CH ₃ ), 2.6-3.6 (m, 6, CH ₂ , SCH ₂ , NCH ₂ ), 4.68 (d of t, 1, H5), 5.07 (s, 2, NCO ₂ CH ₂ ^), 5.28 (s, 2, CO ₂ CH ₂ O), 6.36 (d of q, 1, = CH) and 7.3 (m, 10, ArH);

UV (MeOH) 306 (E 6500 nm;

Mass spectrum m / e 478 (M ⁺ ), 410, 343, 269 and 210.

In an analogous manner, p-nitrobenzyl-6-ethylidene-3- (2-bromo-tert-butyloxycarbonylaminoethyl) -thio-7-oxo-azabicyclo [3.2.0] hept-2-ene-2-carboxylate is obtained, if instead of the N-benzyloxycarbonyl-O-methanesulfonyl-thienamycin benzyl ester from Example 6 the equivalent amount of N-bromotert-butyloxycarbonyl-O-methanesulfonyl-thienamycin-pnitrobenzyl ester of Example 5 used.

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example

Benzyl-ö-ethyl-3- (2-benzyloxycarbonylaminoethyl) -thio ^ -oxo- 1 -azabicyclo [3.2.0] hept-2-en-2-carboxylate ___

LJ

.CO ₂ CH ₂ C ₆ H ₅

A mixture of 15 mg of benzyl 6-ethylidene-3- (2-benzyloxycarbonylaminoethyl) -thio-7-oxo-i-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, 10 mg of Bishop PtOp and 1 , 5 ml of ethyl acetate is hydrogenated in a glass bomb for 2 hours at 2.8 bar (40 psi). Another catalyst (5 mg) is then added and the hydrogenation is continued for a further 2 hours. The mixture is then filtered through a packed layer or bed of 1: 1 silica gel G / MgSO ^ (1 g of each material) using 20 ml of ethyl acetate as the eluent. When the ethyl acetate is evaporated in vacuo, the oil (10 mg) remains, which is purified by thin layer chromatography on a 250 μm 15 cm silica gel GF plate using ethyl acetate / chloroform (3: 1) as the developing agent. The main UV-visible band at R _£ 0.67 is separated and eluted with ethyl acetate. This gives 4 mg of benzyl 6-ethyl-3- (2-benzyloxycarbonylaminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate in the form of an oil with the following characteristics:

IR (CHCl ₃ ) 3470, 1776, 1715, 1505, 1325, 1276, 1230 and

NMR (CDCl ₃ ) Cf 1.01 (t, 3, CH ₂ CH ₃ ), 1.77 (m, 2, Cg ₂ , CH ₃ ), 2.8-3.6 (m, 7, CH ₂ , SCH ₂ , NCH ₂ , H6), 4.19 (d of t, H5), 5 _f 09 (s, 2, NCO ₂ CH ₂ JZf), 5.27 (ABq, 2, CO ₂ CH ₂ O) ~ and 7.32 (m, 10, ArH);

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UV (MeOH) 316 mn;

Mass spectrum m / e 480 (M ⁺ ), 410.

P-Nitrobenzyl-6-ethyl-3- [2- (bromo-tert-butoxycarbonylamino) -thio-7-oxo-1-azabicyclo is obtained in an analogous manner [3.2.0] hept-2-en-2-carboxylate, if instead of benzyl-6-ethylidene-3- (2-benzyloxycarbonylaminoethyl) -thio-7-OXO-1-azabicyclo [3.2.0] hept-2 en-2-carboxylate from Example 7 the equivalent amount of p-nitrobenzyl-6-ethylidene-3- (2-bromo-tert-butyloxycarbonylaminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate begins.

Example 8

6-Ethylidene-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.O] -hept-2-en-2-carboxylate

COOH

10 mg of benzyl 6-ethylidene-3- (2-benzyloxycarbonylaminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate are dissolved in 4 ml of ethanol and the solution is mixed 2 ml 0.1 M pH-7 phosphate buffer and 20 mg 10biger palladium-activated carbon (Pd / C). The mixture is then hydrogenated for 1 hour at 25 ^° C. under atmospheric pressure and then filtered. The filtrate is concentrated in vacuo to 2 ml and then extracted with ethyl acetate. The aqueous portion is chromatographed on a Dowex-50 χ 4 (sodium form) containing ion exchange resin using deionized water as the eluent. "The desired fractions are combined and freeze-dried," 6-Ethylidene-3- (2-aminoethyl) -thio is obtained -7-oxo-1-azabicyclo [3.2cO] hept-2-ene-2-carboxylate.

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Example 9

6-Ethyl-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.O] -hept -2-en-2-carboxylic acid

A / V

/ 7

-COOH

A mixture of 10 mg of benzyl 6-ethyl-3- (2-benzyloxycarbonylaminoethyl) thio-7-oxo-i-azabicyclo [3.2.0] hept-2-ene-2-carboxylate and 20 mg of 10% palladium Activated charcoal in 4 ml of dioxane / H ₂ O (1: 1) is hydrogenated for 1 hour at 2.8 bar (40 psi). The catalyst is then filtered off from the mixture and washed with further HpO (4 ml). The filtrate is combined with the washing water, concentrated in vacuo down to 2 ml and the solution is applied to a column containing Dowex 50x4 ion exchange resin (sodium form) with a particle size of 37 to 74 μm (200 to 400 mesh), which one eluted with deionized water. The appropriate fractions are combined and freeze-dried. 6-Ethyl-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] · hept-2-en-2-carboxylic acid is obtained.

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Example 10

Preparation of sodium 6-ethyl-3- [2- (bromo-tert-butoxycarbonylamino) -ethylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

0 CH ₃ ^^ N- rV-SCH-CH-NHCOC CH-Br

CH-

CO-Na

20 mg of p-nitrobenzyl-6-ethyl-3- [2- (bromo-tert-butoxycarbonylamino) -ethylthio] -7-oxo-1-azabicyclo [3.2.0] -hept-2-en-2- carboxylate in 4 ml of ethanol and 2 ml of 0.1 M phosphate buffer (pH 8.0) and 60 mg of 10% palladium-activated carbon are added to the solution. The mixture is stirred for 75 minutes at 25 ^° C. and 1 atm. H ₂ and then filtered off the catalyst. The filtrate is neutralized with 2.5N HCl (up to pH 7.0), concentrated to 3 ml and extracted briefly with 5 ml of ether. The aqueous solution obtained in this way contains sodium 6-ethyl-3- [2- (bromo-tert-butoxycarbonylamino) ethylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylate .

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example

11

Preparation of 6-ethyl-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid

CO ₂ Na

SCH ₂ CH ₂ NH ₂ ,

λ ^{N L C00H}

The solution obtained according to Example 10, which sodium 6-ethyl-3- [2- (bromo-tert-butoxycarbonylamino) ethylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-en-2 Contains carboxylate, is ^{heated to 60 to 65 0} C for 1 hour. The mixture is then chromatographed on an ion exchange column containing Dowex 50x8 (sodium form), eluting with water. This gives the desired 6-ethyl-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid.

example

12th

N- (p-Nitrobenzyloxycarbonyl) -thienamycin-p-nitrobenzyl ester

140 mg (0.51 mmol) of thienamycin are dissolved in 35 ml of ice-cold water, and 428 mg (5.1 mmol) of sodium bicarbonate are added to the solution. 35 ml of dioxane are then added with stirring and cooling with an ice bath. After 3 minutes, a solution of 165 mg (0.765 mmol) of p-nitrobenzyl chloroformate in 2 ml of dioxane is added dropwise over the course of 2 minutes. The mixture obtained is stirred for 10 minutes in the cold and then extracted with ice-cold diethyl ether (2 × 20 ml). The aqueous phase is separated off, covered with 35 ml of ice-cold ethyl acetate and acidified to pH 2.3 _{with 1 m H 2 SO ^ in an ice bath while stirring vigorously.}

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The layers are then separated from one another and the aqueous portion is extracted with more cold ethyl acetate (2-5 ml). The combined ethyl acetate extracts are with washed ice-cold sodium chloride solution and then extracted thoroughly with 10 ml of 0.05N aqueous LiOH. The LiOH extract is evaporated on a rotary evaporator to separate the ethyl acetate and then freeze-dried. You get in this way 288 mg of crude N- (p-nitrobenzyloxycarbonyl) thienamycin lithium salt in the form of a light yellow pest.

A portion (250 mg) of the crude lithium carboxylate and 373 mg (1.73 mmol) of p-nitrobenzyl bromide in 2.65 ml of anhydrous Hexamethylphosphoramide is stirred for 105 minutes at room temperature. The mixture is then diluted with 60 ml of ethyl acetate, washes with water (2 × 50 ml), 5! & Lger sodium bicarbonate solution (25 ml), more water (2 × 25 ml) and sodium chloride solution (25 ml), dry over magnesium sulfate, filtered and the filtrate evaporated in vacuo to a yellow solid (423 mg). This residue will to separate the excess p-nitrobenzyl bromide with Diethyl ether rubbed. The remaining crystals are filtered off and dried in vacuo. One obtains 126 mg of ^ (p-nitrobenzyloxycarbonyl I-thienamycin-p-nitrobenzyl ester in the form of a yellow solid with the following characteristics:

Mp 163.5-165 ° C;

IR (Nujol-Mull) 1773 and 1690 cm "¹;

NMR (DMSO-dg) d "1.15 (d, 3, CH ₃ ), 2.8-3.6 (m, 7, CH ₂ , SCH ₂ , NCH ₂ , H6), 4.0 (m, 2, U5, H8), 5.22 (s, 2, NCO ₂ CH ₂ Ar), 5.40 (ABq, 2, CO ₂ CH ₂ Ar), 7.70 (m, ArH) and 8.27 ( m, ArH).

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Example 13

N- (p-nitrobenzyloxycarbonyl) -0-methanesulfonyl-thienamycin p-nitrobenzyl ester

CO "CH" - // \\ - N0

An ice-cold solution of 186 mg (0.32 mmol) is added N- (p-Nitrobenzyloxycarbonyl) -thienamycin-p-nitrobenzyl ester in 9.5 ml of anhydrous tetrahydrofuran with stirring with 89 μΐ (0.64 mmol) of triethylamine and then dropwise within 2 min. With a solution of 55 mg (0.48 mmol) methanesulfonyl chloride in 1 ml tetrahydrofuran. The received The mixture is stirred in the cold under nitrogen for 90 minutes and then diluted with 50 ml of ice-cold methylene dichloride. The solution obtained is mixed with 20 ml of cold water, 20 ml of cold 0.1 M pH 3 phosphate buffer and 20 ml of cold 5 # iger Washed sodium bicarbonate solution, dried over magnesium sulfate and filtered. When the filtrate is evaporated in vacuo, a yellow foam is obtained, which can be mixed with anhydrous Diethyl ether rubs. 192 mg of N- (p-nitrobenzyloxycarbonyl) -0-methanesulfonyl-thienymycin-pnitrobenzyl ester are obtained in the form of a light yellow solid with the following characteristics:

Mp 131 Ms 133 ° C;

IR (CH ₂ Cl ₂ ) <fi 786, 1727, 1522 and 1350 cm "¹;

NMR (DMSO-d ₆ ) 1.42 (d, 3, CH ₃ ), 2.5-3.4 (m, 7, CH ₂ , SCH ₂ , NCH ₂ , H6), 3.25 (s, 3 , CH ₃ SO ₂ ), 3.93 (m, 1, H5), 5.03 (m, 1, H8), 5.20 (s, 2, NCO ₂ CH ₂ Ar), 5.43 (m, 2, CO ₂ CH ₂ Ar), 7.63 (m, ArH) and 8.25 (m, ArH);

Mass spectrum m / e 664 (M ⁺ ), 578, 500, 415, 372, 304.

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0.9

Analysis C ₂₇ H ₂₈ N ^ O ₁₂ S ₂ :

C HNS

calc .: 48.79 4.25 8.43 9.65

found: 49.19 4.36 8.09 9.61

Example 14

p-Nitrobenzyl-6-ethylidene-3- [2- (p-nitrobenzyloxycarbonylamino) ethylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate

A solution of 50 mg (0.075 mmol) of N- (p-nitrobenzyloxycarbonyl ) -O-methanesulfonyl-thienamycin-p-nitrobenzyl ester in 0.75 ml of anhydrous dioxane, 2.5 ml of anhydrous methanol and 12.6 mg (0.15 mmol) of sodium bicarbonate are added. The mixture obtained is stirred for 17 hours at room temperature. The precipitate formed is then filtered off, washed out with several portions of water and vacuum dried. 32 mg of p-nitrobenzyl-6-ethylidene-3- [2- (p-nitrobenzyloxycarbonylamino) ethylthio] -7-oxo-1-azabicyclo are obtained [3.2.0] hept-2-en-2-carboxylate in the form of a white solid with the following characteristics:

Mp 160-161 ° C;

IR (CH ₂ Cl ₂ ) 1779, 1730, 1524 and 1351 cm "¹;

NMR (DMSO-d ₆ ) 1.86 (d, 3, CH ₃ ), 2.8-3.8 (m, 6, CH ₂ , SCH ₂ , NCH ₂ ); 4.85 (m, 1, H5), 5.22 (s, 2, NCO ₂ CH ₂ Ar), 5.42 (ABq, 2, CO ₂ CH ₂ Ar), 6.44 (m, 1, = CH), 7.68 (m, ArH) and 8.25 (m, ARH); Mass spectrum m / e 500 (M ⁺ -68), 415, 304, 256, 239, 223, 209.

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example

. 33

6-Ethyl-3- (2-aminoethyl) -thio-7-oxo-1-azabicyclo [3.2.O] -hept -2-en-2-carboxylic acid

A mixture of 37 mg p-nitrobenzyl-6-ethylidene-3- [2- (pnitrobenzyloxycarbonylamino) ethylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate, 40 mg PtO _2, 4 ml of tetrahydrofuran and 2 ml of ethanol is hydrogenated for 2 1/2 hr. at 25 "C and a pressure of 3.5 bar (psi 50). It is then filtered off the catalyst and rinse with 5 ml of 0.1M pH The filtrate combined with the washing water is evaporated in vacuo to separate off most of the organic solvents. The aqueous residue is chromatographed on a column containing Dowex 50 χ 4 ion exchange resin (sodium form), the product being treated with deionized water The correct fractions are combined and freeze-dried, giving 6-ethyl-3- (2-aminoethyl) -thio-7-oxo-T-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid.

example

16

O-sulfo-thienamycin

Th

0-SO ₃ -Na

NH ₂

COOH

A solution of 50 mg of N- (p-nitrobenzyloxycarbonyl) -thienamycin-p-nitrobenzyl ester in 0.5 ml of pyridine is mixed with 20 mg of pyridine-sulfuric anhydride. The mixture is left to ^{react for 4 hours at 25 °} C. and the excess pyridine is then evaporated off under reduced pressure. One stirs the

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Residue 30 min. With 20 ml each of methylene dichloride and 0.1% sodium bicarbonate solution. Then the watery ge layer separated and freeze-dried. The solid residue, which is the sodium salt of O-sulfo-N- (p-nitrobenzyloxycarbonyl) -thienamycin-p-nitrobenzyl ester contains, is dissolved in 20 ml of aqueous dioxane (1: 1). It is then hydrogenated for 4 hours in the presence of 5 mg of platinum oxide as a catalyst at a pressure of 2.8 bar (40 psig). The catalyst is then filtered off and the filtrate extracted twice with ethyl acetate. The watery ge layer is concentrated to 5 ml and chromatographed on 40 ml of XAD-2 resin. The column is eluted with water and that containing the O-sulfo-thienamycin sodium salt Fraction recovered and freeze-dried.

Example 17
Manufacture of medicines

Such a single dosage form is obtained by adding 120 mg of 3- (2-aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.Ojhept-2-en-2-carboxylic acid mixed with 20 mg lactose and 5 mg magnesium stearate and the mixture (145 mg) in a Fill in gelatine capsule number 3. Similarly, using a higher amount of active ingredient and a lower amount of lactose other preparations can be put in gelatin capsules no.3. If required, more than 145 mg Mixing the components together, you can also make larger capsules as well as compressed tablets and pills. the The following examples are intended to explain the manufacture of pharmaceuticals;

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Tablet formulation ^ mg per tablet

3- (Aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid 125

Corn starch, U.S.P. 6th

Dicalcium phosphate 192

Milk sugar, U.S.P. 190

The "active ingredient" is mixed with the dicalcium phosphate, the milk sugar and about half of the corn starch. The mixture
is granulated with 6 mg 15-6 corn starch paste and that
Granules are roughly ^{sieved out, dried at 45 °} C. and sieved again through No. 16 sieve. Then you add the rest of the corn starch and the magnesium stearate and press them
Mixture into tablets, each 1.27 cm (0.5 in.) In diameter and 800 mg in weight.

Parenteral., Solution

ampoule

3- (2-Aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.0 jhept-2-en-2-carboxylic acid 500 mg

Diluent: sterile water for

Injections 2 ml

Eye solution

3- (2-Aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.0] hept-2-en-2-carboxylic acid 100 mg

Hydroxypropyl methyl cellulose 5 mg

sterile water. . 1 ml

Ear solution

3- (2-Aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.0 Jhept-2-en-2-carboxylic acid 100 mg

Benzalkonium chloride 0.1 mg

sterile water. . 1 ml

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Local ointment

3- (2-Aminoethylthio) -6-ethyl-7-oxo-1-azabicyclo [3.2.0Jhept-Z-en-Z-carboxylic acid

Polyethylene Glycol 4000 U.S.P. Polyethylene Glycol 400 U.S.P.

100 mg 400 mg 1.0 g

The active ingredient of the above formulations can be used alone or in combination with other biologically active substances, e.g. other antibacterial agents such as lincomycin, a Penicillin, Streptomycin, Novobiocin, Gentamicin, Neomycin, Colistin or Kanamycin, or with other therapeutic Means such as probationary oath may be administered.

For the availability of microorganisms to which reference is made in the patent (registration) citations mentioned above:

Quote

P 27 24 56O
(USSN
733 655)

P 28 05 701
(USSN
767 723)

P 28 05 724
767 ^S 92O)

US Γο
3,950,357
= German
P 25 52 638

description

Streptomyces cattleya

Protaminobacter ruber (Merck name MB-3528)

Streptomyces flavogriseus

Deposit number / position

NRRL 8057 / ARS Culture Collection, Peoria, 111., U.S.A.

NRRL B-8143 / ARS

NRRL 8139 / ARS

Streptomyces NRRL 11 020 / ARS

flavogriseus

(MA-4638)

MB-3528 as above-

Streptomyces NRRL 814O / ARS

flavogriseus

(MA-46OOa)

MA-4638 as above

MB-3528 as above

MA-46OOa as above

Streptomyces

cattleya

(MA-4297) NRRL 8057 as above

End of description

809842/0898

Claims (9)

DR.-ING. "WALTER ABITZ BR. DIETER F. MORF DIFL.-PHYS. M. GRITSCHNEDER Patentanwälte ΙΛϋηϋιβη, Postal Address Postfach 86Ο1Ο9, 8ΟΟΟ Munich 86 Fienzenauerstraße 28 Telephone 98 32 22 Telegrams: Chemindua Munich Telex: CO) 523993 16011Y PATENT CLAIMS 1. Compound of formula. .N. SCH ₂ CH ₂ NH ₂
COOH and their pharmacologically acceptable salts. 2. Compounds of the general formula CH ₃ CH. s? .N- .COOR in which R ¹ and R each represent a hydrogen atom or an easily removable protective group. 3. A compound according to claim 2, characterized in that R ¹ and R each represent a hydrogen atom. 4. Compounds of the general formula ' - 1 - 809847/0898 RfGlMAL INSPECTED 16011Y α in which R 'and R each represent a hydrogen atom (but R ¹ and R are not simultaneously hydrogen atoms) or an easily removable protective group. 5. Process for the preparation of the compounds according to claim 1, characterized in that a compound is used the general formula CH ₃ CH μ I COOR in which R and R ¹ each represent a hydrogen atom or an easily cleavable protective group, is reacted with a reducing agent. 6. Process for the preparation of the compounds according to claim 2, characterized in that a compound the general formula OR " " * _ COOR in the OR "is an easily removable group and R ¹ and R each represent a hydrogen atom or an easily removable protective group, is reacted with a base. 7. The method according to claim 6, characterized in that a compound is used in which R "is a SuIfo- or organosulfonyl group. - 2 - 16011Y 8. Medicament containing a compound according to claim 1 in addition to the usual auxiliaries and / or packaging agents. 9. A medicament containing, in unit dosage form, a therapeutically effective amount of a compound according to claim 1 in addition to the usual auxiliary and / or assembly aids. 0 RB ι, 2/0 ζ «1 8