DE2652676A1 - N-ALKYLATED DERIVATIVES OF THIENAMYCINE, THE PROCESS FOR THEIR MANUFACTURING AND MEDICINAL PRODUCTS CONTAINING THESE - Google Patents

Description

Dr. \ ηη. YVr ^ r Abüz

Dr. The ^ rF. Morf _ig> November 1976

Dr. Hsns-A. Brauns 15833Y

8 Künsiiiii SJ, P.'anzonauarstr. 28

MERCK & CO., INC.

126 East Lincoln Avenue, Rahway, N.J. Ο7Ο65,

V. St.A.

N-alkylated derivatives of thienamycin »process too their manufacture and medicinal products containing them

The invention relates to certain N-alkyl, Ν, Ν-Dlalkyl- and Ν, Ν, Ν-trialkyl derivatives of the new antibiotic Thienamycin. Such compounds and derm pharmaceutical acceptable salt, ester and amide derivatives are effective as antibiotics.

The invention also relates to a process for the preparation of these compounds and to pharmaceutical compositions containing these compounds. Also describe processes by which such Verbindun- _F gene and their preparations can be administered when an antibiotic effect is indicated.

Thienamycin (I) is described and claimed in US Pat. No. 3,950,357t) This patent is hereby incorporated into US Pat Revelation included, vreil thienamycin as starting

+) (= DT-OS 25 52 638)

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compound in the preparation of the compounds according to the invention can be used:

OH

<f

COOH

Other convenient starting materials for the N-alkylated Thienamycin of the present invention are shown below (Ia, Ib and Ic):

OR-

JCOOH

Yes

COXR

Ib

OR "

-N-

COXR

Ic

wherein R, X and R have the meaning given below to have. Starting compounds Ia, Ib and Ic, which are also called Antibiotics are useful are described and claimed in US patent application Ser. 634 006,

709822/1020

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298 and 63 ¹ J 29 ^, all filed on November 21, 1975 and the corresponding continuation-in-part filings. These registrations have the internal names 15821Y, 15818Y and 15822Y. These applications are hereby incorporated into the disclosure because they describe useful starting compounds and also processes for converting the N-alkylated thienamycins of the present invention to the carboxyl, 0, and carboxyl and 0-derived forms, which are also of the present invention Invention and which are also useful as antibiotics.

The N-alkylated thienamycin derivatives according to the invention can be characterized by the following general structural formula:

OH

I!

-SCH ₂ CK ₂ NR ⁵ R ⁶ R ⁷ COOM

II

or more simply by the symbol:

ΓΟΗ
Th-J-NR ⁵ R ⁶ R ⁷

L COOM

II

where "Th" symbolizes the bicyclic nucleus of thienamycin and the OH, amino and carboxyl groups of thienamycin are given;

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M is H, a salt cation selected from alkali or alkaline earth metals or an amine salt and R, R and R are independently selected from the group consisting of hydrogen (not all R, R and R are hydrogen at the same time) and substituted and unsubstituted residues:

Lower alkyl of 1 to 10 carbon atoms, alkenyl of 2 to 10 carbon atoms, alkynyl of 2 to 10 carbon atoms; ring-substituted and unsubstituted radicals: cycloalkyl, cycloalkenyl, cycloalkenylalkyl and cycloalkylalkyl with 3 to 6 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain, aryl with 6 to 10 carbon atoms, aralkyl with 6 to 10 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain, mono- and bicyclic heteroaryl and heteroaralkyl containing h to 10 ring atoms, one or more of which is selected from the group consisting of oxygen, nitrogen and sulfur and 1 to 6 carbon atoms in the alkyl chain; and wherein the ring or chain substituent is selected from: halogen such as chlorine, bromine, iodine, fluorine, azido, cyano, amino, mono-, di- and trialkyl-substituted amino, wherein the alkyl has 1 to 6 carbon atoms; Hydroxyl, alkoxyI of 1 to 6 carbon atoms; Alkylthioalkyl of 1 to carbon atoms; Carboxyl; Oxo; Alkoxy / carbonyl having 1 to 6 carbon atoms in the alkoxyl group; Acyloxy of 2 to 10 carbon atoms; Carbamoyl and mono- and dialkylcarbamoyl, in which the alkyl group has 1 to * J carbon atoms; Cyanothio (-SCN) and Nitro. It is understood that the Ν, Ν, Ν-trialkyl derivatives are quaternary ammonium compounds and the counter-ion (anion) is not critical and can be selected from halides, such as chlorine and bromine, phosphates, sulfates and the like.

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The compounds according to the invention also encompass embodiments of the following structure:

rOR ³

Th - NR ⁵ R ⁶ R ⁷
.-COXR
II

where the basic symbols have the meanings given above and X is oxygen, sulfur or NR ¹ (R '= H or R) and R can be selected, for example, from the group consisting of hydrogen, customary blocking groups, such as trialkylsilyl , Acyl and the pharmaceutically acceptable salt, ester and amide groups as they are known for bicyclic β-lactams in antibiotics. The definition of R is given in great detail below.

R means: 1.) Acyl (in general the group OR referred to as an ester group); or 2.) R is selected from alkyl, aryl, alkenyl, aralkyl and the like- (see above, that the group OR can generally be called ether). R can also mean hydrogen. Of the The term "acyl" is intended to include by definition the alkanoyles including derivatives and analogues thereof, such as the thio analogs, in which the carbonyl oxygen is replaced by sulfur, as well as sulfur and phosphoric acyl analogs, such as substituted sulfonyl, sulfinyl and sulfenyl radicals, and substituted P (III and V) radicals, such as substituted phosphorous, phosphorus, hypophosphorous and phosphinig residues. Such acyl groups of the present invention are further defined, as are the groups (2nd, above), which is the ether embodiment of the present Represent invention.

- 5 -709822/102 9

There is a constant need for new antibiotics. Because unfortunately there is no static effectiveness for a given antibiotic because it is ongoing Wide application of such an antibiotic selectively causes the formation of resistant strains and pathogens elevated. In addition, the known antibiotics have the disadvantage that they are only effective against certain types of microorganisms are effective. That is why research into new antibiotics continues.

Unexpectedly, it has now been found that the compounds of the present invention are effective against antibiotics Spectrum that are useful in both animal and human therapy and in inanimate systems.

It is therefore an object of the present invention to provide a new class of antibiotics, which have the basic core structure of thienamycin, but which are characterized by the fact that they Are N-alkylated derivatives thereof. These antibiotics are active against a wide range of pathogens for which well1 Gram-positive bacteria are representative, such as S. aureus, Strep, pyogenes, and B. subtilis, and gram-negative Bacteria such as E. coli, Proteus morganii, Klebsiella, are representative. Further objects of the invention are chemical processes for the production of such To provide antibiotics and their non-toxic pharmaceutically acceptable salts, as well as pharmaceutically acceptable ones Compositions containing such antibiotics. The invention also describes methods of treating in which such antibiotics and compositions are administered when an antibiotic effect is indicated.

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at

With regard to the general description of the present invention (structure II, above), the most preferred embodiments are those in which R, R and R are selected from hydrogen, lower alkyl and alkenyl having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, allyl and like; Benzyl and ring-substituted benzyl such as p-tert-butylbenzyl and the like; Heteroaralkylene, such as ¹ I-pyridylir.ethy 1, 2-purylmethyl, 2-thieny! Methyl and the like and R, X and R have the meaning given above - particularly preferred embodiments are those in which R ^]
Mean hydrogen.

those in which R is hydrogen, X is oxygen and R

The compounds according to the invention are obtained by one thienamycin or a suitable derivative thereof or a sufficiently protected thienamycin species with an N-alkylating agent implements. In the procedure according to the present There are no particularly critical conditions in the invention and a large number of the known N-alkylation processes can be used be applied. The identity of the N-alkylating agent is dependent on the choice within

5 6 7

the limits imposed by the definition of R, R and R given are. The N-alkylation can be carried out in a number of ways of solvent systems which are inert or essentially inert to the desired course of the reaction, made will. Suitable solvents include a polar solvent such as water, lower alkanoyl such as Ethanol, dioxane, tetrahydrofuran (THF), acetonitrile, hexamethylene phosphoramide (HMPA), dimethylformamide (DMF) and the like and mixtures (especially aqueous mixtures) of the above solvents; as well as non-polar solvents such as benzene and halogenated hydrocarbons, such as methylene chloride, chloroform and the like.

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VS

Typischervreise the reaction is carried out at temperatures in the range from - ¹ to +50 JO ⁰ C for 15 minutes to 5 hours. In general, the reaction is carried out in the presence of an acid acceptor such as propylene oxide, magnesium oxide, potassium carbonate and the like. The preferred N-alkylating agents include active halides, sulfate esters and Michael addition reagents. The following reagents are representative of such alkylating agents: methyl iodide, allyl bromide, bromoacetone, phenacyl bromide, benzyl bromide, ethyl chloroacetate, propargyl bromide, 2-bromoethyl ethyl ether, dimethyl sulfate and methyl 1-fluorosulfonate, chloromethylthiocyanate, 2-chloromethyl-sulfonate, chloromethylthiocyanate, chloromethyl-pano-methyl-sulfonate, 2-chloropropyl-2-pyromethyl-2, 4-methylpyromethane, chlorobenzyloiditromethyl , Methyl acrylate, nitroethylene and the like.

The monoalkyl embodiments of the present invention can be made in a variety of different ways. A convenient starting material is tris-trimethylsilyl thienamycin (Th (TMS),). if desired

3 3

is that R, R or R · and Reine have other meanings than hydrogen, then suitable derivatives of the starting materials, such as Ia, Ib or Ic (above), can be used. The reaction is carried out in any of the non-protic solvents enumerated above in the presence of the N-alkylating agent of choice (R ¹ X ', where R ^f = R, R or R' and X 'is halogen or sulfate). The desired product is obtained by aqueous hydrolysis following the N-alkylation stage. The following reaction diagram outlines this process:

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τ-OTMS Th _ _NHTMS - COOTMS

Th (TMS) ο

R ¹ X ¹

hydrolysis

Th -

-OTMS NR ¹ TMS .COOTMS

raw

Th --NHR ¹
-COOH

wherein TMS, R ¹ and X ^{1 have} the meaning given above.

A second scheme for the preparation of the monoalkyl embodiments includes the N-alkylation of N-substituted thienamycin in which the substituent is an easily removable, large group (R ⁰ ), such as an aralkyl group, for example a substituted or unsubstituted group such as : Benzyl, benzylhydryl (-CH (CgHj. ) " And trityl (-C (CsR -) -,) where the ring substituent on the aralkyl is halogen, nitro, lower alkyl, lower alkoxyl, and the like. The following reaction diagram outlines this scheme:

Th--

-OR

NHR °
LCOXR

R ¹ X ¹

Th -

-OR

NR ⁰ R ¹ LCOXR

H2 / cat.

Th

rOR ³ NR ¹ -COXR 3

in which all symbols have the meaning given above.

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With regard to the above diagram, the starting material j., Prepared for example from a reaction of thienamycin or a derivative thereof with an aralkyl halide, reacted with the N-alkylating agent of choice R'X ', as previously described, using the N, N-dialkyl intermediate 2 receives. The aralkyl-N-substituent R ° can be easily removed, obtaining "5" by hydrogenolysis. Suitable conditions for this final cleavage stage include the hydrogenation of 2 in a solvent such as ethanol under hydrogen (1 to ¹ J atmospheres) in the presence of a Catalyst, such as platinum, palladium or oxides thereof. The end product of this reaction is finally 3, the N-N-Ntono-lower alkyl. However, the Ν, Ν-diene-lower alkyl-thienamycin is present together. Such contaminating by-products can be separated by chromatographic processes and the scope of the Contamination can be reduced by using 1 equivalent or less of the alkylating agent R ¹ X '- · - ■

A third process for the preparation of the N-monoalkyl species, particularly the N-lower alkyl species, is similar the procedure described above with the exception, that the starting material la is Ν, Ν-diaralkyl-thienamycin. The preparation of this starting material is described below. The following reaction diagram outlines this Procedure:

"OR ³ rOR ³ T-OR ³

Th -NR ⁰ R ⁰ th - NR ⁰ R ⁰ R ¹ Th.-NR ¹

LCOXR

-COXR

-COXR

JLa
in which all symbols have the meaning given above.

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It should be noted that this scheme for the preparation of N-lower alkyl-thienamycins is not complicated is made by the coproduction of Ν, Ν-di-lower alkyl-thienamine.

A fourth process which is particularly suitable for the preparation of N-lower alkyl thienamycin species includes the N-alkylation of a Schiff's base of thienamycin. The following diagram shows this reaction:

r OR ³ rOR ³ ", -OR ³

¹ X ¹

-COXR

MR

Hydrolysis or

Th - N = C-0 ^> Th - N = C-0 -— ~ - _: ^ Th-.N-R '

T _T Ii λ hydrogenation

.COXR

COXR 0 _ _phenyl

All symbols have the meaning given above and in addition 0 denotes phenyl, R and R can be trimethylsilyl radicals and X can be oxygen. The preferred Schiff base is that obtained by reacting thienamycin with benzaldehyde or a ring-substituted benzaldehyde. In this process it is not critical how the Schiff base was prepared and its preparation is described in US patent application Serial Mo. 63 ^ 297 and its continuation-in-part application, which has the internal file number 15833IA US patent application is included here for disclosure purposes because it describes the preparation of the starting compound H. The reaction of Ji ₁ with an alkylating agent R'X 'gives the intermediate product 5, which in an aqueous hydrolysis or catalytic hydrogenolysis, the desired N-lower alkyl thienamycin species 6 gives.

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A fifth method for producing N-lower alkyl thienamycin includes desulfurization of one N-thioacyl-thienamycins in the presence of a hydrogenation catalyst, such as Raney nickel:

Th-

-OR ³

-L-NKCR ⁸ _Th .

¹ S S

-OR ³

-NHCH ₂ R '

-COXR LCOXR

where mean: X is oxygen, R and R have the before given meaning, but preferably mean water

substance, and R = hydrogen, aryl or a lower alkyl radical with 1 to 5 carbon atoms. The N-thioacylthienamycin starting material is fully described in U.S. patent application serial no. 63 ** 291 of November 21 1975 and the corresponding continuation-in-part registration, which has the internal file number 15775Y and which is hereby incorporated into the disclosure because it is the manufacture of the starting material concerns. The aforementioned desulfurization is typically carried out in a polar protic solvent such as water, lower alkanol, such as ethanol, or aqueous mixtures thereof at temperatures between 0 and 50 C during the period of 30 minutes to 5 hours.

The N, N-dialkyl-thienamycin derivatives according to the invention can be made in a variety of ways. The N-substituents can be identical ("symmetrical") or be different ("asymmetrical"). The symmetrical type can be made from the starting compound 1 with an excess of the alkylating agent used. In this process, only the M, N-disubstituted is obtained Product.

- 12 -

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The hydrogenation of the resulting intermediate to Cleavage of the N-aralkyl substituent gives the desired one N-dialkyl-thienamycin species in that previously described Way.

In general, the N, N-dialkyl-thienamycin species can be prepared by direct alkylation, when the alkylation reaction is carried out in water, a protic solvent, or a mixture thereof, without protecting the carboxyl group. However, if this direct alkylation is carried out in an aprotic solvent such as HI4PA, esterification of the free carboxyl group generally takes place. In those cases in which ester formation is not desired, the carboxyl group is preferably blocked by a customary, easily removable carboxyl blocking group. (Preparation of such carboxy1-blocked thienamycin species is described later.) It should also be noted that direct N-alkylation of the thienamycin generally yields a mixture of products consisting of the mono-, di- and trialkyl species . The relative proportions result from steric reasons, which are determined by the size of R ^{1 of} the alkylating agent and the amount of reagent used. If R ^{1 is} small (less large), such as methyl or ethyl, the Ν, Μ, Ν-trialkyl species predominate. If the size of R 'increases, the mono- and dialkyl species predominate.

The symmetrical type can either be produced by reductive alkylation with an aldehyde (R "CHO,

5 6 7

wherein -CHpR "= R, R or R) or by transalkylation of an ester of thienamycin by means of an alkyl halide

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15 83-3 Y

or sulfate and, if desired, subsequent cleavage the ester group by standard procedures such as a hydrogenation.

The asymmetric types can be prepared by reductive alkylation of a monoalkylthieneamycin species with a suitable aldehyde or by alkylation of tris-trimethylsilyl-N-monoalkylthienamycin with an alkylating agent of choice in the presence of an acid acceptor such as propylene oxide, K ₃ CO ^ or MgO . Again, any starting materials Ia, Ib or Ic can be used to obtain the corresponding 0-, carboxyl or 0- and carboxyl derivatives. The following diagram describes the aforementioned reactions:

OTMS

Th - "R ¹ TMS -COOTMS

R ¹¹ X ¹

Acid acceptor

hydrolysis

-OTMS Th __NR'R "

-COOTMS -OH

Th - NR ¹ R "_COOH

wherein all symbols have the meaning given above and R ^{f can be the} same or not the same R "and where R 'and R" are selected from R ⁵ , R ⁶ and R ⁷ ;

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15833

Th.

r OR

■ ^ΝΗ

R ¹ X '

MgO

_Hv drifting H ₂ / Pd

-OR

Th __N (R ') ₂

-COOR 3

-OR Th - N (R ^f } -COOH

Herein, R has the meaning given previously and provides in the case of arr-circuit takes place cleaving a readily removable blocking group such as benzyl or p-nitrobenzyl; R has the meaning given above, R '= R ⁵ , R ⁶ or R ⁷ ;

-OR

Th J-NH

1.) R ¹¹ CHO

2.) E ₂ / Pt

^L -COOR

-OR

Th - N (CH ₂ R ") ^ COOR

Herein -CH ₂ R "has the meaning R ⁵ , R or R ⁷ and R and R have the meaning given above.

-OR

Th-. NH

L COOR

R ¹ X ¹

HMPA

r OR

Th - N (R ') -COOR

or

ρ OR Th-- NH

2 L COOH

R ¹ X '

HMPA

OR

Th-JN (R ') -, LCOOR ¹

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Here, R has the meaning given above; R '=

CD 7

R, R or R '; X ¹ = halide or sulfate; R has the meaning given - the special case in which R = H has been described here. Again, it should be noted here that ester formation can be avoided when working with the free acid if the alkylation is carried out in water or a protic solvent.

Ν, Ν, Ν-trialkyl-thienamycin derivatives of the present invention may be selected from thienamycin, an O-, carboxyl- or O- and carboxyl derivative thereof, or from the N, N-dialkyl species by alkylation with an alkyl halide or sulfate. The carboxyl group can are protected by a common biocide group such as Benzyl or p-nitrobenzyl. It should be recorded here, however that R- and R-substituted N, N-dialkylthienamycins Valuable substrates are, if desired, compounds of the present invention of the following Making structure:

Th

-OR "

Wr ⁵ R ⁶ R ⁷

-COXR

where the non-critical counter anion A was previously has been identified. The following diagram reflects this last procedure:

Th-

OTMS KR ⁵ R ⁶

L-COOTMS

1.) R ⁷ X '

2 ·) hydrolysis

-OH

3 5 6 7 ThXNR R R

_COC

wherein all substituents are given above.

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IH

Identification of the remainder -COX'R

In the general description of the compounds (Ha) according to the invention, the radical denoted by the group -COX ¹ R means, inter alia, -COOH (X is oxygen and R is hydrogen) and all radicals which are known to be effective pharmaceutically acceptable esters , Anhydrides (R is acyl) and amide residues in bicyclic ß-lactaman antibiotics, as in the case of cephalosporins and penicillins and their essentially analogues.

Suitable radicals (R) include customary protective or Carboxy blocking groups. The term "blocking group", as used herein is applied in the same manner as that disclosed in US Pat. No. 3,697,515, which is hereby incorporated by reference with respect to the disclosure is used. Pharmaceutically acceptable thienamycin derivatives

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of the present invention falling under this class are given below. Suitable blocking esters include probes such as those in the following List can be given, which is representative but by no means an exhaustive list of the possible Represents ester groups in which X = O and R are given:

(i) R = CR ^a RR ^C , where at least one of R ^a , R and R is an electron donor, for example p-methoxyphenyl, 2,4,6-trimsthylphenyl, 9-anthryl, methoxy, CH "SCH" tetrahydrofuran 2-yl, tetrahydropyran-2-yl or

from c

Fur-2-yl. The remaining R, R and R groups can be hydrogen or organic substitution groups. Suitable Estergruppen this type include pT * ethoxybenzyloxycarbonyl and 2, ⁱ i, 6-Triitiethylbenzyloxycarbonyl.

(ii) R = CR ^a RR ^c , wherein at least one of the R ^a , R and R groups is an electron attractive group, for example benzoyl, p-nitrophenyl, JJ-pyridyl, trichloromethyl, T rib r ο nine thy 1, iodomethyl , Cyanonethyl, ethoxycarbonylmethyl, arylsulphonylir.ethyl, 2-dimethy1sulphoniummethyl, o-nitrophenyl or cyano. Suitable esters of this type include benzoyl methoxycarbonyl, ρ-Ni t rob en zyloxy carbonyl, ¹ I-pyridyl methoxy carbonyl, 2,2,2-trichloroethoxycarbonyl and 2,2,2-tribromethoxycarbonyl 1.

(iii) R = CR R R, viorin at least two of the R, R and R radicals are hydrocarbons, such as alkyl, for example

709822/1029

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i (o OR ζ 9 R 7 PJ

wise Ksthy1 or ethyl or aryl, for example phenyl and the remaining R, R and R ^c group, if present, is hydrogen. Suitable esters of this type include tert-butyloxycarbonyl, tert-amyloxycarbonyl, diphenyl methoxycarbonyl and triphenylmethoxycarbonyl.

(iv) R = R, where R is adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl or tetrahydropyran-2-yl.

Silyl esters of this category of blocking groups can can be produced in a simple manner from halosilanes or silazanes of the formula:

R ² LSiX ¹ ; R * LsiX '; R ^, Si.Nr \ ,; R ¹ LSi.NH.COR *; R -Si.NH.CO.NH.SiR,; R NH.CO.NH .SiR,; or RC (OSiR,); HN (SiR,) _OJ in which X ¹ denotes halogen, chlorine or bromine and the various groups R, which can be identical or different. Hydrogen atoms or alkyl, for example methyl, ethyl, n-propyl, isopropyl; aryl, for example phenyl; or aralkyl, for example a bensyl group.

More generally, pharmaceutically acceptable carboxy derivatives of the present invention are those derived by reacting N-acylated thienamycin (1) with alcohols, phenols, mercaptans, thiophenols, acylating agents and the like to give compounds (2, above), which are then converted to derivatives to give the R group of the compounds of the present invention (II, above). For example, interesting esters and amides are the compounds of the formula II (above) with the following groups in the 2-position: -COXR, where X is oxygen, sulfur or NR '(R ¹ = H or R),

709822/1029

and R is alkyl with 1 to 10 carbon atoms, straight or branched, such as methyl, ethyl, tert .. butyl, Pentyl, decyl and the like; Carbonylmethyl, including Phenacyl, p-bromophenacyl, tert-butylphenacyl, Acetoxyacetylmethyl, pivaloxyacetylmethyl, carboxymethyl and its alkyl and aryl esters, oC-carboxy-oC-isopropyl; Aminoalkyl including 2-methylaminoethyl, 2-diethylaminoethyl, 2-acetamidoethyl, Ph tha III mi dome thy 1, succinic acid imidone ethyl; Alkoxyalkyl, in which the alkoxy part has 1 to 10 and preferably 1 to 6 carbon atoms and is branched, can be straight-chain or cyclic, and wherein the alkyl part has 1 to 6 carbon atoms, such as methoxymethyl, Ethoxymethyl, isopropoxymethyl, decyloxymethyl, ethoxypropyl, Deeyloxypentyl, cyclohexyloxymethyl and the like; Alkanoyloxyalkyl, wherein the alkanoyloxy part is straight-chain or branched and has 1 to 6 carbon atoms and the alkyl part Has 1 to 6 carbon atoms, such as acetoxymethyl, pivaloyloxymethyl, acetoxyethyl, propionyloxyethyl, acetoxypropyl and the same. Haloalkyl, in which halogen is chlorine, bromine, fluorine or iodine and the alkyl part is straight-chain or is branched and has 1 to 6 carbon atoms, for example 2,2,2-trichloroethyl, trifluoroethyl, 2-bromopropyl, Diiodomethyl, 2-chloroethyl, 2-bromoethyl and the like; Alkenyl with 1 to 10 carbon atoms, which is either straight-chain or branched, for example allyl, 2-propenyl, 3-butenyl, 4-butenyl, i | -pentenyl, 2-butenyl, 3-pentenyl, 3-methyl-3-butenyl, Methallyl, 1,4-cyclohexadien-1-yl-methyl and the same; Alkynyl of 1 to 10 carbon atoms which can be either straight-chain or branched, for example 3-pentenyl, propargyl, ethynyl, 3-butyl-l-yl and the-

- 20 ^

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same; Alkanoyl, which can be either straight-chain or branched, with 1 to 10 carbon atoms ₃ such as pivaloyl, acetyl, propionyl and the like; Aralkyl or heteroalkyl, in which the alkyl has 1 to 3 carbon atoms, and hetero means that 1 to b heteroatoms are present, selected from the group consisting of 0, S or N, such as benzyl, benzhydryl and substituted benzyl, benzhydryl, for example benzyl or benzhydryl substituted with 1 to 3 substituents such as benzyl, phenoxy, halogen, lower alkyl, lower alkanoyloxy with 1 to 5 carbon atoms, lower alkoxy. Hydroxy, nitro, blocked carboxy or combinations thereof, for example p-chlorobenzyl, o-nitrobenzyl, 3,5-dinitrobenzyl, p-methoxybenzyl, m-benzoylbenzyl, p-tert.-butylbenzyl, m-phenoxybenzyl, p-benzoylbenzyl, p- Nitrobenzyl, 3,5-dichloro- ⁱ -hydroxybenzyl, p-methoxycarbonylbenzyl, p-methoxybenzhydryl, p-carboxybenzyl, the latter either being a free acid, an ester or a sodium salt, 2,4,6-trimethylbenzyl, p- Pivaloyloxybenzyl, p-tert-butoxycarbonylbenzyl, p-methylbenzyl, p-benzoyloxybenzyl, p-acetoxybenzyl, p-2-sthylhexanoylbenzyl, p-ethoxycarbonylbenzyl, p-benzoylthiobenzyl, p-benzamidobenzyl, p-benzamidobenzyl, p-benzamidobenzyl, o -benzylloxy, p-benzamidobenzyl, o -benzylloxy-iso-benzamidobenzyl, p-benzamidobenzyl, o -benzylloxy, p-benzamidobenzyl, p-benzamidobenzyl, o -benzylloxy, m-benzamidobenzyl, o -benzylloxy-p-benzamidobenzyl, p-benzoyloxybenzyl, o -benzylloxy , p-tert-butoxybenzyl, as well as the cyclic analogues thereof, 2,2-dimethyl-5-cumaranmethyl, 5-indanylmethyl, p-Trimethylsilylbenzyl, ^{3,5-bis-t-butoxy-2} J-hydroxybenzyl; 2-thienylmethyl, 2-furylmethyl, S-tert-butyl-S-isothiazolmethyl, 6-pivaloyloxy-3-pyridazinylethyl, 5-phenylthio-ltetrazolymethyl or the like (the term lower alkyl or lower alkoxy in this context being a chain with 1 to h is carbon atoms); or phthalidyl; or phenylethyl, 2- (p-methylphenyl) ethyl and the

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Arylthioalkyl analogs, aryloxyalkyl, in which aryl is preferably a phenyl ring with 0 to 3 substituents, preferably 0 or 1 substituents in the ortho or para position and the alkyl has 1 to 6 carbon atoms, for example (I-methoxy) -phenoxymethyl, phenoxymethyl, ( Ij-chloro ^ phenoxymethyl, ( ^J 4-nitro) -phenoxymethyl,

(4-Benzyloxy) -phenoxymethyl, (1-methyl) -phenoxymethyl, (2-methoxy) -phenoxymethyl, (1-phenoxy) -ethyl, (4-amino) -phenoxymethyl, (1-methoxy) -phenyl thiome thy I. , ( ¹ I-ChIOr) -phenylthiomethyl, phenylthioethyl; Aryl, in which aryl is phenyl, 5-indanyl or substituted phenyl with 0 to 3 substituents, preferably 0 to 1 substituents in the ortho or para position, for example (4-methyl) -phenyi, (* 1-hydroxy) -phenyl , (4-tert-butyl) -phenyl, p-nitrophenyl, 3,5-dinitrophenyl or p-carboxyphenyl, v; whether the latter is present either as a free acid or as a sodium salt; aralkenyl, in which aryl is phenyl or alkenyl with Means 1 to 6 carbon atoms, such as 3-phenyl-2-propenyl; Aralkoxyalkyl, in which aralkoxy is benzyloxy and the alkyl has 1 to 3 carbon atoms, such as benzyloxymethyl, (^ -nitrobenzyloxymethyl, (4-chloro) -benzyloxymethyl; alkylthioalkyl, viorin the alkylthio part has 1 to 10 and preferably 1 to 6 carbon atoms, but that branched, straight-chain or cyclic and the alkyl part has 1 to 6 carbon atoms, such as methylthioethyl, ethylthioethyl, cyclohexylthiomethyl, decylthiobutyl, methylthiopropyl, isopropylthioethyl, methylthiobutyl and the like ..

In addition to the esters (and thioesters) that have been listed above, amides are also used by the

709822/1029

- 22 -

lying invention includes, that is, in which X

R ¹
is an -N group. Typical examples of such amides are those in which R ^{1 is} selected from the group e consisting of hydrogen, methyl, ethyl, phenyl, p-methoxyphenyl. Benzyl, carboxyir.ethyl, methylthioethyl and heteroaryl; The grouping -COXr also includes anhydrides in which R is acyl, for example benzyloxycarbonyl, ethoxy carbonyl, benzoyl and pivaloyl.

The most preferred -COXR- radicals of the present invention are those in which, relative to structure II, X is oxygen, sulfur, or NR ¹ (R 'is selected from the group consisting of hydrogen and lower alkyl); and R is selected from the group consisting of: lower alkyl, lower alkenyl such as methallyl, 3-methylbutenyl, 3-butenyl, and the like; Methylthioethyl; Ber.zyl and substituted benzyl, such as p-tert-butylbenzyl, m-phenoxybenzyl, p-pivaloyloxybenzyl, p-nitrobenzyl and the like; Pivaloyloxynethyl, 3-phthalidyl and acetoxymethyl, propionyloxymethyl, acetylthiomethyl, pivaloylthiomethyl, allyl, ^ l-butenyl, 2-butenyl, 3-methyl-2-butenyl, phenacyl, acetoxyacetylmethyl, methoxymethyl, poxyacetylmethyl, poxy-poxy-poxybenzyl, pivalo-acetoxybenzyl, pivalo-acetoxybenzyl , 5-indanylmethyl, 5-indanyl, benzyloxymethyl, ethylthioethyl, methylthiopropyl, methoxycarbonyloxymethyl, xthoxycarbonyloxymethyl, dimethylaminoacetoxymethyl, crotonolacton-3-yl and acetamidomethyl.

709822/1029 - 23 -

MA

Identification of R

In a general presentation of the present invention In structure II, the radical R means except hydrogen 1.) Acyl (in general the group becomes -OR referred to as ester); or 2.) R is selected from alkyl, aryl, aralkyl, and the like, making the group -OR can be called an ether. For the ester version (1) R is selected from the following definitions for acyl radicals (p = 1). In the so-called ether execution (2.) of the present invention, R is selected from the same acyl radicals in which the carbonyl-0 . ν

group -C- or generally -C- is omitted (p = O); so that R is selected from the following radicals, for which all symbols are designated below:

C / - (CH ₂ ) _n ZR * -VC / -CER "

Therefore, in relation to the definition of R, the acyl radical including a substituted or unsubstituted aliphatic, aromatic or heterocyclic, be araliphatic or heterocyclylaliphatic carboxylic acid residue, a substituted or unsubstituted carbamyl radical or a carbothioic acid radical. A group of acyl residues can be represented

709822/1029

are given by the general formula:

-C-R "

where X = O or S and R "means hydrogen; amino; substituted amino, such as alkyl and dialkylamino, in which the alkyl radical comprises 1 to 6 carbon atoms; substituted or unsubstituted radicals, such as: straight-chain or branched-chain alkyl radicals, in which the alkyl radical 1 to Comprises 6 carbon atoms; mercapto, aryloxy, which typically comprises 6 to 10 carbon atoms; alkenyl or alkynyl groups, which typically comprise 2 to 6 carbon atoms; aryl, such as phenyl; aralkyl, such as benzyl; cycloalkyl, which typically contains 3 to 6 carbon atoms or a heteroaryl or heteroaralkyl group (mono- or bicyclic) wherein the alkyl group typically contains 1 to 3 carbon atoms and the heterocyclic ring typically contains H to 10 atoms and the heteroatom or atoms are selected from 0, N and S; the The aforementioned groupings can be unsubstituted or they can be substituted by radicals such as OH, SH, SR (R is low alkyl or aryl, such as phenyl), alkyl or alkoxy groups having 1 to about 6 carbon atoms, halogen, such as Cl, Br, R and J, cyano, carboxy, sulfamino, carb, amoyl, sulfonyl, azido, amino, substituted amino, such as alky Amino including quaternary ammonium, in which the alkyl groups have 1 to 6 carbon atoms, haloalkyl, such as trifluoromethyl, carboxyalkyl, carbamoylalkyl, N-substituted carbamoylalkyl, in which the alkyl group

- 25 _

709822/102 9

15833Y

of the aforementioned four radicals comprises 1 to about 6 carbon atoms, amidino, guanidino, N-substituted guanidino, guanidino-lower alkyl and the like. Typical examples of such acyl groups which can be mentioned here are those in which R "means benzyl, p-hydroxybenzyl, ¹ J-AmLnO- ¹ I-carboxybutyl, methyl, cyanomethyl, 2-pentenyl, n-amyl, n- Heptyl, ethyl, 3 ~ or ^ -nitrobenzyl, phenethyl, ß, ß-diphenylethyl, methyldiphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2,6-dimethoxyphenyl, 2,4,6-trimethoxyphenyl, 3,5-dir _; ethyl- ¹ i-isoxazolyl, 3-butyl-5 ~ methyl-JJ-isoxazolyl, 5-methyl-3-phenyl- ¹ I-isoxazolyl, 3- (2-chlorophenyl) -5-r: ethyl- ⁱ l-isoxazolyl, 3- (2,6-DichlorphenyD-S-methyl - ^ - isoxazolyl, D-il-Amino - ^ - carboxybutyl, D- ^ N-Benzoylamino - ^ - carboxy-n-butyl, p-aminobenzyl, o-aminobenzyl, m- Aminobenzyl, p-dimethylaminobenzyl, (3-pyridyl) methyl, 2-ethoxy-1-naphthyl, 3-carboxy-2-quinoxalinyl, 3- (2,5-dichlorophenyl) -5- (2 ~ furyl) -4- isoxazolyl, 3-phenylil-isoxazolyl, 5-methyl-3 ~ ( ⁱ * -guanidinophenyl) - ⁱ l-isoxazolyl, H- guanidinomethylphenyl, 4-guanidinomethylbenzyl, 4-guanidinobenzyl, 4-guanidinophenyl, 2,6-diinethoxy- 4-guanidi no, o-sulfobenzyl, p-carboxymethylbenzyl, p-carbamoylraethyl- · benzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-haphthylmethyl, 3-isothiazolylmethyl, 4-isothiazolylmethyl, 5-isothiazolyl , ^ -Pyridylmethyl, 5-Isoxazolylmethyl, 4-Methoxy-5-isoxazoly! ^{Methyl, 1} I-Me thy 1-5- is oxazolylmethyl 1, 1-Imidazo IyI-methyl, 2-Benzofuranylmethyl, 2-Indolylmethyl, 2- Phenylvinyl, 2-phenylethynyl, 1-aminocyclohexyl, 2- and 3-thienylaminomethyl, 2- (5-nitrofuranyl) vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl, o-pheny! Phenyl, p-aminomethylbenzyl,

~ 26 - 709822/1029

2657676

1- (5-Cyanotriazolyl) methyl, Di fluorine thy 1, dichloromethyl, dibromomethyl, 1- (3-MethyIimidazοIyI) methyl, 2- or 3- (5-carboxymethylthienyl) methyl, 2- or 3- (4- Carbamoylthienyl) methyl, 2- or 3- (5-methylthienyl) methyl, 2- or 3- (Methoxythienyl) methyl, 2-or 3 ~ (IJ-chlorothienyl) methyl, 2- or 3- (5- Sulfothienyl) methyl, 2- or 3- (5-carboxythienyl) methyl, 3- (1,2,5-thiadiazolyD-methyl, 3- ( ¹ <-methoxy-1,2,5-thiadiazolyl) methyl, 2-Furyimethyl, 2- (5-nitrofuryl) methyl, 3-Furyimethyl, 2-thienylmethyl, 3-thienylmethyl, tetrazolylmethyl, benzamidinomethyl and cyclohexylamidinomethyl.

The acyl group can also be a radical of the formula:

X
I!

-C (CH ₂ ) _n ZR "

wherein X is 0 or S and η is 0 to 4 and Z Is oxygen, sulfur, carbonyl or nitrogen and R ″ has the meaning given above. Typical Members of the substituent

- (CH ₂ ) _n ZR »,

which should be mentioned here are allylthiomethyl, Phenylthiomethyl, Butylmereaptomethy1, oL-Chlorocrotylmercaptomethyl, Phenoxymethyl, phenoxyethyl, phenoxybutyl, phenoxybenzyl, diphenoxymethyl, dimethylmethoxyethyl, Dimethylbutoxymethyl, dimethyl phenoxy methyl, jj-guanidinophenoxymethyl, Ij-pyridylthiomethyl, p- (carboxymethyl) -

709822/1 029 '" ²⁷

15833Y

phenoxymethyl, p- (carboxymethyl) -phenylthiomethyl, 2-thiazolylthiomethyl, p- (sulfo) -phenoxymethyl, p- (carboxymethyl) -phenylthiomethyl, 2-Pyrimidinylthiomethyl, PhenäthyIthiomethy1, l- (5,6,7,8-tetrahydronaphthyl) oxomethyl, N-Kethyl-4-pyridylthio, benzyloxy, methoxy, ethoxy, phenoxy, Phenylthio, amino, methylamino, dimethylamino, pyridinium methyl, Trimethylammonium methyl, cyanomethylthiomethyl, trifluoromethylthiomethyl, ll-pyridylethyl, i | -pyridylpropyl, ^ -Pyridylbutyl, 3-imidazolylethyl, 3-imidazoIyI-propyl, 3-imidazolylbutyl, 1-pyrroloethyl, 1-pyrrolopropyl and 1-pyrrolobutyl.

Alternatively, the acyl group can be a radical of the formula

-C-CHR "
ι

R "'

where R "has the meaning given above and R" ^{1 is} a radical such as amino, hydroxy, azido, carbamoyl, guanidino, amidino, acyloxy, halogen, such as chlorine, fluorine, bromine, iodine, sulfamino, tetrazolyl, Sulfo, carboxy, carbalkoxy, phosphono and the like. Typical members of the substituent

- CHR "

which should be mentioned here are oC-aminobenzyl, oC-amino- (2-thienyl) -methyl, oC- ( methylamino) -benzyl, OG Ami η ome thy 1-

709822/1029

- 28 -

Wed ₀

mercaptopropyl, oC-amino-3- or -ll-chlorobenzyl, ^ 3- or -il-hydroxybenzyl, oC-amino-2,4-dichlorobenzyl, cC-amino-3 _s ⁱ »-dichlorobenzyl, D (-) - OG -Hy droxybenzyl, oC ^ oxybenzyl, tf? -Amino- (3-thienyl) -methyl, D (-) - o? -Ainino-3-chloro - ^ - hydroxybenzyl, oC- amino- (cyclohexyD-methyl, o £ (5-Tetrazolyl) -benzyl, 2-Thieny lea rh oxy line thy I, 3-Thienylcarboxymethyl, 2-Fury! carboxymethyl, 3-Fürylcarboxymethyl, oC-Sulfamlnobenzyl, 3-Thienylsulfaminomethyl, c / - (N- Methy isulfamino) benzyl, D (-) - 2-thienylguanidinomethyl, di-i-of-guanidinobenzyl, oC-guanylureidobenzyl, oC-hydroxybenzyl, cG-azidobenzyl, oC-pluobenzyl, 4- (5-methoxy-l, 3-oxadiazolyl) aminomethyl, ^ - (5-i ^y fethoxy-l, 3-oxadiazolyl) -hydroxymethyl, 4- (5-methoxy, 3-sulfadiazolyl) -hydroxymethyl, 4- (5-chlorothienyl) aminomethyl, 2 - (5-Chlorothienyl) -hydroxymethyl, 2- (5-chlorothienyD-carboxymethyl, 3- (1,2-thiazolyl) -aminomethyl, 3- (1,2-thiazolyl) -hydroxymethyl, 3- (1,2-thiazolyl ) -carboxymethyl, 2- (1,4-thiazolyl) -aminomethyl, 2- (1,4-thia2olyl) - hydroxymethyl, 2- (1,4-thiazolyl) -carboxymethyl, 2-benzroi; hienylaminomethyl, 2-benzothienyl, hydroxymethyl, 2-benzothienylcarboxymethyl, o ^ -sulfobenzyl, oG-phosphonobenzyl, oC-diethylphosphono and oC-monoethylphosphono.

Other interesting acyl groups in this class where X = oxygen are:

»3 it
-CCHR R,

3 4. 3

wherein R and R are defined below. R means hydrogen, halogen, such as chlorine, fluorine, bromine, iodine, amino, Guanidino, Phosphono, Hydroxy, Tetrazolyl, Carboxy, SuIfο or sulfamino and R denotes phenyl, substituted

_ 29 _
709822/1029

Phenyl, a mono- or bicyclic heterocyclyl containing one or more oxygen, sulfur or nitrogen atoms in the ring, such as furyl, quinoxalyl, thienyl, Quinolyl, quinazolyl, thiazolyl, thiothiazolyl, tetrazo IyI, oxadiazolyl, thiadiazolyl and the like, substituted heterocycles, phenylthio, phenyloxy, lower alkyl with 1 to 6 carbon atoms, heterocyclic or substituted heterocyclic thio groups or cyano.

3 ii

The substituents on the groups R and R can be halogen, carboxymethyl, guanidino, guanidinonethyl, carboxyamidomethyl, aminomethyl, nitro, methoxy or methyl. If R is selected from the group consisting of hydrogen, hydroxy, amino or carboxy and R ^ is selected from the group consisting of phenyl or a 5- or 6-membered heterocyclic ring with one or two sulfur, oxygen or nitrogen heteroatoms, such as tetrazolyl , thienyl, furyl and phenyl, the following acyl radicals are as typical examples: phenylacetyl, 3 ^'i -Bromphenylacetyl, p-Aminomethylphenylacetyl, 4-acetyl-Carboxymethylpheny1, h- Carboxyamidomethylphenylacetyl, 2-furylacetyl, 5-nitro-2-furylacetyl, 3 -Furylacetyl, 2-thienylacetyl, 5-chloro-2-thienylacetyl, 5 ~ methoxy-2-thienylacetyl, oC-guanidino-2-thienylacetyl, 3-thienylacetyl, 2- (* J-methy 1-thienyD-acetyl, 3- Isothiazolylacetyl, ^ -Methoxy - ^ - isothiazolylacetyl, il-isothiazolylacetyl, 3-methyl-4-isothiazolylacetyl, 5-isothiazolylacetyl, 3-chloro-5-isothiazolylacetyl, 3-methyl-1,2,5-oxadiazolylacetyl, 1,2, 5-thiadiazolyl-4-acetyl, 3-methy1-1,2,5-thiadiazolylacetyl, 3-chloro-1,2,5-thiadiazolylacetyl, 3-methoxy-1,2,5- thiadiazoly 1-acetyl, phenylthioacetyl, ^ -pyridylthioacetyl, cyanoacetyl, 1-tetrazolylacetyl, oC-fluorophenylacetyl, D-phenylglycyl, ^ l-hydroxy-D-phenylglycyl, 2-thienylglycyl, 3-thienylglycyl,

- 30
70 9 822/1029

Phenylmalonyl, 3-chlorophenylmalonyl, 2-thienylmalonyl, 3-thienylmalonyl, oC-phosphonophenylacetyl, cC-aminocyclohexadieny! Acetyl, oc ^ sulfaminophenylacetyl , oG hydraxyphenylacetyl, o ^ tetrazolylpheny! Acetyl and c £ -Sulfopyl and c £ -Sulfopyl.

The acyl radical can also be selected from sulfur (I) - and phosphorus (2) residues;

(O) m. (x) n

-S-Y

Il I

(O) η γ

rti

wherein with respect to 1, m and η are integers selected between O and 1 and Y ° = CP Pr, -M (R ") ₂ and R"; where Mr ^ is selected from hydrogen, alkali metal cations and organic bases and R "has the abovementioned meaning, for example alkyl, alkenyl, aryl and heteroaryl. With regard to J ^, X = O or; η = 0 or 1; and Y ' and Y "are selected from the groups consisting of 0 ^Θ κΡ, -N (IT) ₂ , R" and ZR ", in which all symbols have the meanings given above, and, for example, R" and ZR "typically have the following meanings: alkyl , Alkenyl, aryl, heteroaryloxy, Y ¹ and Y ", including the R" groups, which can be linked to one another to form cyclic esters, ester-amide and amide functions. Typical examples of I ^ are 0- (methylsulfonyl) -thienamycin, 0- (p-mitrophenylsulfonyl) -thienamycin, 0- (p-chlorophenylsulfinyl) -thienamycin, 0- (o-nitro-

709822/102 9 - ³¹

15833Y ty

phenylsulfenyl) thienainycin, O-sulfamicin-0-sulfonic acid, O-dimethylsulfamicin-thienamycin and the sodium salt of thienamycin-0-sulfonic acid. Typical examples of 2 ^ are O- (Dimethoxyphosphino) -thienamycin, O-dibenzyloxyphosphino) -thienamycin ₉ 0- (Dihydroxyphosphino) -thienamycin disodium salt, _ 0- (Dimethoxyphosphinyl) -thienamycin, 0- (Dihydroxyphosphino) -thienamycin disodium salt, _ 0- (Dimethoxyphosphinyl) -thienamycin, 0- (Dihydroxyphosphino) -0- (Dihydroxyphosphino) -Dienotharnothiny, 0- (Dihydroxyphosphino) -Dienamycin, 0- (Dihydroxyphosphino) -dienamycin -thienamycin and O- (dihydroxyphosphinyl) -thienamycin disodium salt.

One class of acyls of particular interest are those Acyl radicals selected from the group consisting of from customary known N-acyl blocking or protecting groups, such as carbobenzyloxy, ring-substituted carbobenzyloxy, such as o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, Chloroacetyl, bromoacetyl, pheny! Acetyl, tert-butoxycarbonyl, trifluoroacetyl, broinethoxycarbonyl, 9-pluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichloroethoxycarbonyl, bromo-tert-butoxycarbonyl, Phenoxyacetyl; non-acylated protecting groups, vrie tri-lower alkylsilyl, for example Tri me thy lsi IyI and tert-butyldimethylsilyl are also "

The following radicals are preferred in accordance with the aforementioned definition of acyl:

Formyl, propionyl, butyryl, chloroacetyl,

Methoxyacetyl, aminoacetyl, methoxycarbonyl, ethoxycarbonyl, Methylcarbamoyl, ethylcarbamoyl, phenylthiocarbonyl, 3-aminopropionyl, 4-aminobutyryl, N-methyl-

709822/102 9 - 32 -

1583 3Υ

5Ό

aminoacetyl, Ν, Ν-dimethylaminoacetyl, Ν, Ν ₃ N-trimethy1-aminoacetyl, 3- (N, N-dimethyl) -aminopropionyl, 3- (N, N, N-trimethyl) -aminopropionyl, N, N, N- Triäthylaminoacety1, Pyridiniumacetyl, Guanidinoacetyl, 3-Guanidinopropionyl, N -Methylguanidinopropionyl, hydroxyacetyl, 3-hydroxypropionyl, acryloyl, Propynoyl, malonyl, phenoxycarbonyl, Amidinoacetyl, Acefeamidinoacetyl, Amidinopropionyl, Acetamidinopropionyl, Guanylureidoacetyl, Guanylcarbamoyl, Carboxymethylaminoacetyl, Sulfoacetylaminoacetyl, Phosphonoace ty ty laminoace 1, N -Dime thy laininoacet ami dinopropionyl, ureidocarbonyl, dimethylaminoguanylthioacetyl, 3- (l-methyl-4-pyridinium) -propionyl, 3- (5-aminoimidazoll-yl) -propionyl, 3-methyl-l-imidazoliuinacetyl, 3-sydnonylacetyl o-aminomethylbenzoyl, o-aminobenzoyl _a sulfo, Phos phono,

-P (OCH ₃ J ₂ ,

fi 9

Il Il ■ | i

-P [N (CH ₃ ) ₂ ] -PN (CH) -P [N (CH)]

-PN (CH ₃ J ₂

ONa

709822/1029 " ³³

15833.Y

SA

A particularly preferred class of acyl residues are terminal substituted acyls in which the substituent is basic Group that may be substituted or unsubstituted can: amino, amidino, guanidino, guanyl and nitrogen-containing mono- and bicyclic heterocycles (aromatic and non-aromatic) in which the hetero atom or atoms are selected in addition to nitrogen from oxygen and sulfur. Such substituted acyls can be represented by the following formula:

O
-C (CH ₂ ) -A- (CH ^) ₁

are characterized in which m and η are integers from 0 to 5;

A is 0, NR ¹ (R ¹ is hydrogen or lower alkyl of 1 to 6 carbon atoms), S or A is a single bond; and Y is selected from the following groups:

1 ») Amino or substituted amino:

-M (R) ₂ and -N (R) ₃ ,

wherein the values for R are independently selected from: hydrogen; N (R ') ₂ ( ^Rl is hydrogen or lower alkyl of 1 to 6 carbon atoms); Lower alkyl and lower alkoxy 1 having 1 to 6 carbon atoms; Lower alkoxy lower alkyl wherein the alkyl group has 1 to 6 carbon atoms and the alkyl group has 2 to 6 carbon atoms;

709822/1029

■ si ·

Cycloalkyl and cycloalkylalkyl, in which the cycloalkyl group Comprises 3 to 6 carbon atoms and the alkyl group Has 1 to 3 carbon atoms, and where two R groups can be connected to one another via the N atom to which they are bound to form a Ring with 3 to 6 atoms.

2.) Amidino and substituted amidino:

-N = CN (R) ₂
. R.

wherein the values for R are independently selected from the groups: hydrogen; N (R ^r ) _? (R * is hydrogen or lower alkyl of 1 to 6 carbon atoms); Lower alkyl and lower alkoxyl of 1 to 6 carbon atoms, lower alkoxy lower alkyl in which the alkoxy group contains 1 to 6 carbon atoms and the alkyl group contains 2 to 6 carbon atoms (if the lower alkoxy lower alkyl radical is bonded to one carbon atom, the alkyl group contains 1 to 6 carbon atoms); Cycloalkyl and cycloalkylalkyl wherein the alkyl group contains 1 to 3 carbon atoms; two R groups can be bonded to one another with the atom to which they are bonded to form a ring with 3 to 6 atoms;

3.) Guanidino and substituted guanidino:

-NH-CN (R) ₂ ,
NO

where R has the definition given under 2. above.

709822/1029

- 35 -

15833Y

• 5 V

M.) Guanyl and substituted guanyl:

-C = NR
NO )

where R has the definition as given in 2 above.

5.) Nitrogen-containing mono- and bicyclic heterocyclyls (aromatic and non-aromatic) with h to nuclear atoms, in which the heteroatom or the heteroatoms other than nitrogen are selected from oxygen and sulfur. Such heterocyclyls are typically illustrated by the following list of radicals (R ¹ is H or lower alkyl having 1 to 6 carbon atoms)

709822/1029

15833Y

NH ¹

-R

- / tfR

709822/1029

- 37 -

The following specific Acylrecte, aie in this class are also representative and are preferred: ^.

O NH

HI

CCH ₂ CH ₂ NHC-CH ₃

NH

CCH ₂ CH ₂ NHC-H

O NH

Il I

-CCH ₂ CH ₂ NHC-NH ₂

Ii

-CCH ₂ CH ₂ N (CH ₃ )

-CCH ₂ CH ₂ N (CH ₃ ) ₃ O NH

Il I

-CCH-CH-C-NH, O NH

Il I

-CCh ₂ CH ₂ CH ₂ NHC-CH ₃

-CCH ₂ CH ₂ CH ₂ N CCH ₃ ) O

II

-CCH ₂ CH ₂ CH ₂ N (CH ₃ ) O

I -CCH-SC

-CCH ₀ S-CH ₀ -C ^

O l

I ^

-CCH ₂ -O-CH ₂ C

709822 / 1.029 ^

- 38 -

15833Y 265 7 676

It will be understood, however, that any acyl radical can be used in the practice of the invention and can be incorporated into the Invention is included.

Production of the starting materials Ia, Ib and Ic

The previously described starting materials are simply prepared from an N-protected thienamycin (1) such as an N-acylated thienamycin (I) ₅

Τ "OH

L COOH
1

1 2
wherein R and R are selected from hydrogen and the aforementioned acyl radicals. Preferably R is

Hydrogen and R is an easily removable blocking group, such as: carbobenzyloxy, ring-substituted carbobenzyloxy, such as o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, Chloroacetyl, bromoacetyl, phenylacetyl, t-butoxycarbonyl, trifluoroacetyl, bromoethoxycarbonyl, 9-fluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichloroethoxycarbonyl, bromo-t-butoxycarbonyl, Phenoxyacetyl; non-acyl-protected groups, such as tri-lower alkylsilyl, for example trimethylsilyl and Tertiary butyldimethylsilyl are also of interest. The on Most preferred N-blocking groups are substituted and unsubstituted carbobenzyloxy radicals:

R = - C - OCH

709822/102 9

where n is 0 to 2 (η = O, R ¹ = hydrogen) and R 'is lower alkoxy or nitro and bromo-tert-butoxycarbonyl.

The final N-deblocking in the Preparation of Ia, Ib or Ic is carried out by a variety of known methods, including Hydrolysis or hydrogenation; when hydrogenation is used, suitable conditions include solvent one such as a lower alkanoyl in the presence of a hydrogenation catalyst such as palladium, platinum or oxides thereof.

The N-acylated intermediate / 1, above 7 is produced

made by thienamycin (I) with an Acylierungsir.ittel treated, for example an acyl halide or an acyl anhydride, such as an aliphatic, aromatic, heterocyclic, araliphatic or heterocyclic aliphatic carboxylic acid halide or anhydride. Other Acylating agents that can also be used are for example mixed carboxylic acid anhydrides and in particular lower alkyl esters of mixed carboxylic acid anhydrides; also carboxyic acids in the presence of a carbodiimide, such as 1,3-dicyclohexylcarbodiimide, and an activated one Are esters of carboxyic acids such as p-nitrophenyl esters suitable.

These N-acylated thienamycin precursors become full described in US patent application serial number

709822/1020

15833Y

291 of November 21, 1975 and the continuation-in-part US application, which was filed simultaneously with the present application for the same applicant, with the internal designation 15775 ^γ Α. These registrations are hereby included in the description.

The acylation reaction can be carried out at temperatures in the range from about -20 to about 100 C, - but temperatures in the range from -9 ° C. to 25 ° C. are preferably used. Any solvent in which the reactants are soluble and which is essentially inert can be used, for example polar solvents, viie Water, alcohols and polar organic solvents, generally those such as dimethylformamide (DMP), hexamethylphosphoramide (HMPA), acetone, dioxane, tetrahydrofuran (THF), acetonitrile, heterocyclic amines, such as Pyridine, ethyl acetate, aqueous mixtures of the aforementioned, as well as halogenated solvents such as methylene chloride and chloroform- The reaction is carried out for a time ranging from about 5 minutes to a maximum of 3 hours, but generally a reaction time of about 0.5 to about 1 hour will range the end. The following equation describes this process using a carboxylic acid halide; However, it is it goes without saying that by using a carboxylic acid anhydride or another functional equivalent Acylating agent can obtain the same products:

709822/102 9

15833Y

Acy! Halogeni d

CH ₂ CH ₂ NRIR COOH

Generally when in the acylation described above reaction an acid halide (suitable halides are chlorides, iodides or bromides) or an acid anhydride is used, the reaction is carried out in water or a aqueous mixture of a polar organic solvent such as acetone, dioxane, THP, DMF, acetonitrile and the like in the presence of a suitable basic acceptor, such as NaHCO ,, MgO, NaOH, KpHPO ^ and the like.

In carrying out the reaction described here, it is generally not necessary to use the 2-carboxy group or to protect the 1'-hydroxy group; in the cases in which the acylating agent is extremely sensitive to water it is sometimes advantageous to carry out the acylation in a non-aqueous solvent system. TriorganosiIyI (or tin) derivatives of thienamycin result quickly the Tris-Triorganosilylderivate, for example Tris-TrimethylsiIy! thienamycin Th (TMS) -:

Th__

- OTMS

NHTMS
L COOTMS

7098 22/10 29

15833Y

These derivatives, which are easily soluble in organic solvents, are easily prepared by one thienamycin with an excess of hexamethyldisilazane and a stoichiometric amount of trimethylchlorosilane at 25 ° C. with vigorous stirring under an N atmosphere implements. The resulting NH Cl is removed by centrifugation and the solvent is passed through Evaporation removed, leaving the desired SiIylderivat receives.

The intermediate starting product Ic is produced according to the following scheme; It should be noted, however, that a direct esterification without protection of the amino group also is possible.

Γ OH

■ V

_IR ^X R ^A > Th-j-HR R

• COOH

1 Ib

raw

Ent-, 1-D ² blocking

raw

-NH ₂ i-COX

in which all symbols have the meaning given above.

In general, the transition (1- ^ Ib) is effected according to customary prior art procedures. These procedures include:

1.) Reaction of 1 (or I) with a DAazoalkane, such as diazomethane, pheny! Diazomethane, diphenyldxazomethane,

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and the like in a solvent such as dioxane, ethyl acetate, acetonitrile and the like, at temperatures between 0 ⁰ C and. Reflux for a period of a few minutes to 2 hours.

2.) Reaction of an alkali salt of 1 with an activated alkyl halide, such as methyl iodide, benzyl bromide or m-phenoxybenzyl bromide, p-tert-butylbenzyl bromide, pivaloyloxymethyl chloride and the like. Suitable reaction conditions include solvents such as hexamethylphosphoramide and the like and temperatures from ⁰ ° C. to 60 ° C. for a period of a few minutes up to 4 hours.

3 ·) Reaction of 1 with an alcohol such as methanol, ethanol, benzyl alcohol and the like. This reaction can be carried out in the presence of a carbodiimide condensing agent such as dicyelohexylcarbodiimide and the like. Suitable solvents at temperatures between 0 C and the reflux temperature and reaction times of 15 minutes to 18 hours include CHCl ^, CIUCl, CHpCl and the like.

k. ) Reaction of an N-acylated acid anhydride of 1, prepared by reacting the free acid 1 with an acid chloride, such as ethyl chloroformate, benzyl chloroformate and the like, with an alcohol such as those listed under 3.), under the same reaction conditions as under 3.) called- The anhydride is made by reacting 1 and the acid chloride in a solvent such as tetrahydrofuran (THP) ₅ CH ₂ Cl ₂ and the like _f

709822/102 »

at temperatures from 25 C to the reflux temperature for 15 minutes to 10 hours.

5.) Reaction of labile esters of 1, such as trimethylsilyl ester, dimethyl-tert-butylsilyl ester or the like, with RX ¹ in which X 'is halogen, such as bromine or chlorine, and R has the meaning given above, in a solvent, such as THP, CHpCl_ and the like at temperatures between 0 C and reflux for 15 minutes to 16 hours. This implementation takes place, for example, according to the following scheme:

rOTMS

Th ₁ -NHTMS N-acylation p ^ _Th _ [. _NR l _TM3 esterunp ^ _Th COOTtfS

-OTKS

Ver

Γ OTiOS

-NR ^- TMS .COOR

mild hydrolysis y

Th- -IiHR ¹ -COOR

wherein TMS is triorganosilyl, such as trimethylsilyl and all other symbols have the meanings given above.

The amides of the present invention are readily prepared by reacting the acid anhydride of Ib (X = O, R = acyl) with ammonia or with the selected amine, for example the aforementioned alkyl, dialkyl, Aralkyl or heterocyclic amines.

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The schemes for the esterification given above are known for the related bicyclic β-lactan antibiotics and in general with organic syntheses and it should be noted here that no special critical conditions with regard to the reaction parameters in the preparation of the N-acylated and carboxy! derivatives Ib, which are used as starting compounds for the present invention are suitable, are required.

The starting compounds Ia and Ic are easier Way prepared by any of the known esterification or etherification reactions of the secondary Alcohol group from Ib. Such procedures include:

"OH J ₀ T?

Th - NR ¹ R ² > Th - NR ¹ R ²

-COXR LcOXR

Ib ■ 2

1.) For the preparation of the ether embodiments of the present invention, the acid-catalyzed reaction of Ib is carried out with a diazoalkane such as diazomethane, phenydiazomethane, diphenydiazomethane and the like in an inert solvent such as dioxane, tetrahydrofuran (THP), halogenated hydrocarbons such as CH ₂ Cl ₃ , ethyl acetate and the like in the presence of a catalytic amount of a strong acid or Lewis acid such as toluenesulfonic acid, trifluoroacetic acid, fluoroboric acid, boron trifluoride and the like ', at temperatures between ~ 78 C and "25 ° C for a few minutes to 2 hours.

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2.) For the preparation of the ether embodiments of the present invention, the reaction of Ib is carried out with an alkylating agent, such as an active halide, for example methyl iodide, benzyl bromide, m-phenoxybenzyl bromide and the like; Alkyl sulfonate such as dimethyl sulfate, diethyl sulfate, methyl fluorosulfonate and the like in the presence of a strong base capable of forming an alcoholate anion of Ib. Suitable bases include alkali and alkaline earth oxides and hydroxides, alkali alkoxides such as potassium tert-butoxide, tert. Amines such as triethylamine, alkali alkyls and aryls such as phenyl lithium, and alkali amides such as sodium amide. Suitable solvents include an inert anhydrous solvent such as tert-butanol, dimethylformamide (DMF) ₃ THP, hexamethylphosphoramide (HMPA), dioxane and the like, at temperatures from -78 ° C. to 25 ° C. for a period of a few minutes to U hours.

3.) For the preparation of the ester embodiments of the present invention, the reaction of Ib with any of the acyl radicals listed above takes place in its acid form. This reaction can be carried out in the presence of a carbodiimide condensing agent such as a dicyclohexylcarbodiimide and the like. Suitable solvents include an inert solvent such as CHCl ,, CH ₂ Cl ₂ , DMF, HMPA, acetone, dioxane and the like, at temperatures from 0 ⁰ C to 6 '
from 15 minutes to 12 hours.

at temperatures from 0 C to 60 C for a period of time

¹ J.) For the preparation of the ester embodiments of the present invention, Ib is reacted with an acyl halide or an acid anhydride, the acyl groups having been previously described. In general one uses at

709822/102

the aforementioned acylation reaction an acid halide (suitable halides are chlorides, iodides or Bromides or acid anhydrides) and the reaction is carried out in an anhydrous organic solvent such as acetone, Dioxane, methylene chloride, chloroform, DMF or the like, in the presence of a suitable basic acceptor, such as NaHCO. ,, MgO, triethylene, pyridine and the like, at temperatures between 0 C to 40 C for one hour to 4 hours.

Suitable acyl halides and anhydrides include: Acetic anhydride, bromoacetic anhydride, propionic anhydride, benzoyl chloride, phenylacetyl chloride, azidoacetyl chloride, 2-thienylacetyl chloride, 2-, 3- and 4-nicotinyl chloride, p-Mitrobenzoylchlorid, 2,6-Dimethoxybenzoylchlorid, 4-guanidinophenylacetylchlorid, Hydrochloride, methanesulfonyl chloride, Dibenzyl phosphorochloridate, dimethylthiophosphorochloridate, 2-furoylethylcarboxylic acid anhydride, methyl chloroformate, Bis (p-nitrobenzyl) phosphorochloridate and the like.

5.) For the preparation of the ester embodiments of the present invention, Ib is reacted with a suitable substituted ketene or isocyanate, such as ketene, dimethyl ketene, methyl isocyanate, methyl isothiocyanate, chlorosulfonyl isocyanate and the like. Suitable solvents include dioxane, tetrahydrofuran, chloroform and the like, at temperatures from -70 ⁰ C to 60 ° C over a period of 15 minutes to 18 hours.

Intermediate ^ j is then N-deblocked as previously described to give starting compounds Ic.

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15833Υ

fcfr

Ia is made from Ic by deblocking the carboxyl group:

-OR ³

Th - 3

-cox R

-> Th-I-NH

^L COXR

Ic

JS «ph JL '

"OR"

^ ₂

.COOK

Yes

The starting material Ia becomes simple and preferable obtained when X is oxygen and R is an easily removable carboxy protecting or blocking group (see above). The starting material Ia is produced by deblocking using a large number of known processes including hydrolysis and hydrogenation. The preferred carboxy blocking groups are used (below) so is the preferred deblocking procedure the hydrogenation, in which the intermediate (Ic or 2) in a solvent such as a lower alkanoyl in The presence of a hydrogenation catalyst such as palladium, platinum or oxides thereof.

In this context it should be noted that suitable "blocking groups" R include the subgroups which are defined as above as aralkyl, haloalkyl, alkanoyloxyalkyl, alkoxylalkyl, alkenyl, substituted Alkyl or aralkoxyalkyl and also alkylsilyl in which the alleyl group has 1 to 10 carbon atoms. Suitable "blocking groups" R include, for example Benzyl, phenacyl, p-nitrobenzyl, methoxy methyl,

709822/1029

Trichloroethyl, trimethylsilyl, tributyltin, p-methoxybenzyl and benzhydryl. These blocking groups are preferred because they are generally recognized as being lightweight removable blocking groups in the prior art relating to the cephalosporins and penicillins.

The preferred carboxy 1 blocking groups are benzyl and substituted benzyl:

where η = 0 to 2 (n = 0, R '= H) and R ^{1 is} lower alkoxyl or nitro.

According to an alternative procedure, the compounds of the present invention can also be obtained by working on the N-alkylated thienamycin to to obtain the derivatives by introducing the group R and / or -COXR. Such a procedure is carried out in the same way as previously described with the exception that the N-alkylated species replaces the N-acylated species, and, of course, there is no need to perform N deblocking.

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The products according to the invention (II) form a large one Number of pharmacologically acceptable salts with inorganic and organic bases, including, for example Include metal salts derived from alkali and alkaline earth hydroxides, carbonates or bicarbonates and Salts derived from primary, secondary or tertiary amines, such as monoalkylamines, dialkylarcines, trialkylamines, Lower alkanolamines, di-lower alkanolardnene, Nie dri galkylenedi amines, N, N-di aralkylnie dri galkylenedi amines, Aralkylamines, amino-substituted lower alkanols, Ν, Ν-di-lower alkylamino-substituted lower alkanols, Amino-, polyamino- and guanidino-substituted Niertrig - alkanoic acids and nitrogen-containing heterocyclic amines. Typical examples include salts that are derived from sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium hydroxide, calcium carbonate, Trimethylamine, triethylamine, piperidine, morpholine, quinine, lysine, protamine, arginine, procaine, ethanolamine, Morphine, benzylamine, ethylenediamine, N, N'-dibenzylethylenediamine, diethanolamine, piperazine, dimethylaminoethanol, 2-amino-2-methyl-1-propanol, thiophylline, N-methylglucamine and the same. Acid addition salts, for example hydrofluoric acid, hydrobromic acid, sulfuric acid, Nitric acid, p-toluenesulfonic acid and methanesulfonic acid can be used in cases where the acyl residue contains a basic group.

The salts can be monosalts, such as the monosodium salts, obtained by reacting one equivalent of sodium hydroxide with one equivalent of product II, or mixed Disalts. Such salts are obtained by adding an equivalent of a base with a divalent cation, such as Calcium hydroxide, with an equivalent of product II

709822/102 9 _ ₅₁

implements. The salts according to the invention are pharmacological acceptable non-toxic derivatives which can be used as active ingredients in suitable single-dose formulations for pharmaceutical products can be used. They can also be combined with other medicines to achieve a wider range of activities.

The new thienamycin derivatives of the invention are valuable Antibiotics with activity against various gram-positive and gram-negative pathogens such as Bacillus subtilis, Salmonella schottmuelleri and Proteus vulgaris. The free acids and especially their salts, such as the Amine and metal salts, especially the alkali and alkaline earth salts, are viable bactericides and can be used to treat harmful bacteria on medicinal products and dental equipment to separate microorganisms and for therapeutic use in humans and animals. For the latter use pharmaceutically acceptable salts min inorganic or organic bases are used, whereby one uses such, which are already known from their use in the administration of penicillins and cephalosporins. For example, salts such as alkali and alkaline earth salts and primary, secondary and tertiary amine salts can be used for this purpose. These salts can be used with pharmaceutically acceptable liquid or solid carriers processed into suitable single-dose formulations such as pills, tablets, capsules, suppositories, syrups, elixirs and the like, these being Administration forms can be prepared according to known methods of the prior art.

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To

The new compounds are new valuable antibiotics which are active against various gram-positive and gram-negative bacteria and which therefore find use in human and veterinary medicine. The compounds of the invention can therefore be used as antibacterial medicaments for the treatment of infections caused by gram-positive and gram-negative bacteria, for example against Staphylococcus aureus, Escherischia coli, Klebsiella pneumoniae, Bacillus subtilis, Salmonella typhosa, Pseudomonas and Bacterium proteus. The bactericides according to the invention can also be used as additives for animal feed and for preserving food and as disinfectants. For example, they can be used in aqueous compositions at concentrations ranging from 0.1 to 100 parts of the antibiotic per million parts of solution to destroy or inhibit the growth of harmful bacteria on medical and dental equipment

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and also as bactericides in industrial applications, for example in paints based on water, in white water from paper mills, to the growth of inhibit harmful bacteria.

The products of the present invention can be used alone or in combination with active ingredients in a variety of pharmaceutical preparations. These Antibiotics and their corresponding salts can be in the form of capsules or tablets, as a powder or as a liquid Solutions or as suspensions and elixirs can be used. You can take it orally, intravenously, or intramuscularly administered.

The compositions are preferably formulated in such a form that they will be absorbed in the gastrointestinal tract will. Tablets and capsules for oral administration may be in unit dosage form and may be conventional Examples such as binders contain, for example, syrup, gum arabic, gelatin, sorbose, tragacanth or polyvinylpyrrolidone; Fillers, for example lactose, sugar, corn starch, calcium phosphate, sorbose or glycerine; Lubricants, for example magnesium stearate, talc, polyethylene glycol, silicon dioxide; Disintegrants, for example Potato starch or suitable humectants such as sodium lauryl sulfate. The tablets can be known in Way to be coated. Oral liquid preparations can be in the form of aqueous or oily suspensions, Solutions, emulsions, syrups, elixirs, and the like, or can be prepared as a dry product so they can then be mixed with water or another

709822/1029 _ 54

suitable carrier material can be prepared before use. Such liquid preparations can be customary Additives, such as suspending agents, for example sorbose, syrup, methyl cellulose, glucose / sugar syrup, Gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, Contain aluminum stearate gel or hydrogenated edible oils, contain, for example, almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; Preservatives, for example methyl or propyl p-hydroxybenzoate or sorbic acid. Contain suppositories the usual substances suitable for this, for example cocoa butter or other glycerols.

Compositions for injections can be given in single doses manufactured in the form of ampoules or in containers with added preservatives for multiple doses. The compositions can be in the form of suspensions, solutions or emulsions in oil or aqueous carrier materials and they can be formulation agents, such as suspending agents, stabilizing agents and / or dispersing agents. Alternatively, the ak tive ingredients are in the form of powder so that mixed them with a suitable carrier material, for example sterile, aseptic water, before use can be.

The compositions can also be in suitable forms be prepared for absorption through the mucous membranes of the nose and throat and bronchial tissues and them can easily be in the form of powdery or

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liquid sprays are prepared for inhalation or in the form of lozenges or preparations for throat brushing. For medical use on the eyes or ears, the preparations can be used as special capsules in liquid or semi-liquid form, or they can be used as drops and the like. For topical Applications, hydrophobic or hydrophilic bases can be formulated as ointments, creams, lotions, anointing agents or powders.

In addition to a carrier material, the inventive Compositions also include other ingredients, such as stabilizers, binders, antioxidants, preservatives, Lubricants, suspending agents, viscosity agents or flavoring agents and the like. Furthermore Other active ingredients may also be present in cen compositions to provide a wider range of antibiotic Effecting activity.

For example, the compositions can be used in veterinary medicine as breast-insertable preparations with either a rapid or a slow release effect be formulated.

The dosages to be administered depend to a considerable extent on the condition of the patient being treated and on the patient's weight, the route of administration and the frequency of administration, the parenteral route being preferred for general infections and the oral route for infections of the bowels. In general

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a daily oral dosage contains about 15 to about 600 mg of active ingredient per kg of body weight of the Patients with one or more administrations per Day. A preferred daily dosage for adults is in the range of about 50 to 120 mg of active ingredient per kg of body weight.

The present compositions can be administered in various unit dosage forms, such as solid or liquid orally ingestible dosage forms. The compositions per single dose, whether liquid or solid, should contain from about 0.1% to 99% active material, the preferred range being between about 10 and 60 % . The compositions generally contain from about 15 mg to about 1500 mg of the active ingredient; however, in general, it is preferred to use a dosage level in the range of about 250 to 1000 mg. In the case of parenteral administration, the unit dose is generally the pure compound in a slightly acidified aqueous sterile solution or in the form of a soluble powder which is intended to be dissolved.

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τ «Ι

The examples below are intended to illustrate the compounds that Further explain the methods and medicaments of the invention; however, they are not to be interpreted in a restrictive sense.

B ice ρ i e 1

Production of N, N-di-p-tert-butylbenzyl-thienamycin-p-tert. butyl benzyl ester

-OH
Th - N (R),

^U CO ₂ R

40 mg of thienamycin are stirred for 3 1/2 hours in 0.6 ml of hexamethylphosphoramide (HMPA) with 0.0275 ml of p-tert-butylbenzyl bromide. The thienamycin goes into solution within 30 minutes. The reaction mixture is then diluted with ethyl acetate and washed successively with aqueous K ₂ HPO., Water (twice) and sodium chloride solution. The ethyl acetate layer is dried over magnesium sulfate, filtered and evaporated. The residue is chromatographed by thin-layer chromatography on silica gel, eluting with 5 ° methanol in CHCl. This gives 4 mg of the pure desired compound (R _f about 0.6); IR (µ, PiIm): 3 »0, OH; 5 »62, β-lactam; 5 * 90, ester; Nuclear Magnetic Resonance (NMR) ( S _{CDCl 5} ): 1.29 (a, t-Bu), 2.6 to 3.3 (m, CH ₂ groups), 3.57 (s, NCH ₂ ), 3.8 to 4.4 (m, β-lactam CH groups), 5.23 (s, OCH ₂ ), 7.29 (s, aryl); MS: m / e 710, 666, 624, 367, 335, 322 (t-Bu = = tert-butyl).

Using the same method, but instead of p-tert.-70 9822/10 2 9 - 58 -

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Butylbenzyl bromide the equivalent amount of p-methoxybenzyl bromide uses, one obtains ΙΙ, Ν-bis-p-methoxybenzyl-thienaniycin-p-methoxybenzyl ester.

example

N-p-nitrobenzyl-thienamycin-p-nitrobenzyl ester and Ν, Ν-bisp-nitrobenzyl-thienamycin-p-nitrobenzyl ester

A solution of 71 mg of thienamycin in 2 ml of dimethyl sulfoxide is mixed with a solution of 27 mg of triethylamine in 0.27 ml
Methylene dichloride and then 56 mg of p-nitrobenzyl bromide were added. The mixture is stirred 30 min. At 25 ⁰ C and then treated it with 7 ml of methylene dichloride and 7 ml of 0.1 N
pH 9 buffer. The organic phase is separated off with
Washed water and sodium chloride solution and evaporated.
The residue is collected at a 20.32 χ 20.32 cm (8 χ 8 in.)
large 1000 μ-310 ₂ -Ρ ^^ β chromatographed, the development being carried out with ethyl acetate. The band or the strip at 0.5 to 2.5 cm is extracted with ethyl acetate. One obtains
the Np-nitrobenzyl-thienamycin-p-nitrobenzyl ester;

267 πιμ Sh (shoulder) 320 ιημ; relative absorbance

ΠΙα, Χ

3: 2. The band at 4.5 to 7 cm provides the N, N-bis-pnitrobenzyl-thienamycin-p-nitrobenzyl ester; UV ^ l 267 πιμ Sh 320 πιμ; relative absorbance 2: 1.

example

Production of N, N-dimethyl-thienamycin

ρ OH
Th - N

-CO ₂ H 12.7 mg of thienamycin are hydrogenated, which is dissolved in 2 ml of 0.07% aqueous

709822/102 9 -59-

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ger formaldehyde solution was dissolved "at a pressure of 2.81 kp / cm (40 psi) in the presence of 14 mg platinum oxide as Catalyst. After 4 hours, the solution is adjusted to a pH of 7 * 0 and it is chromatographed on a 15 ml column containing XAD-2 resin. The column is eluted with water. You catch a faction, which one Contains mixture of thienamycin and N, N-dimethyl-thienamycin. Analytical high pressure liquid chromatography on Cjg-Porasil with 10% tetrahydrofuran in water as solvent gives two peaks "with retention times of 1.75 and 2.5 minutes.

Example 4

0 (TMS)
Th-I-NH (TMS) or Th (TMS)

COO (TMS)
TMS = trimethylsilyl group

Production of silylated thienamycin

80 mg of thienamycin are suspended under nitrogen in 40 ml of tetrahydrofuran, the suspension is concentrated to 10 ml and 1 ml of hexamethyldisilazane and 300 μl of trimethylchlorosilane are added. The mixture is allowed to ^{react for 20 minutes at 25 °} C. with vigorous stirring. The ammonium chloride is then centrifuged off from the suspension and the supernatant is evaporated in a stream of nitrogen to give an oil which is used for the subsequent reaction.

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18 26-S2676

Example 5

Manufacture of N-methyl-thienamine

Th (TMSK prepared from 20 mg of thienamycin is dissolved in 1 ml of tetrahydrofuran, 15 mg of dimethyl sulfate are added to the solution and the mixture is stirred for 2 hours at 23 ^° C. Then 1 ml of phosphate buffer (pH 7) is added and the mixture is made up The mixture is then adjusted to pH 7 and extracted with ethyl acetate, the aqueous layer is separated and applied to a column (40 ml) of XAD-2 resin The column is eluted with water and the outflow is monitored by determining the refractive index and the UV absorbance. The first fractions, which contain inorganic salts, are discarded. The portions containing the N-methylthienamycin are strongly concentrated and freeze-dried.

example N-Me thy 1-thi enamyc in

A mixture of 272 mg of thienamycin and 280 mg of trimethylsilylimidazole in 20 ml of tetrahydrofuran is ^{stirred at 25 °} C. for 30 minutes. Then 120 mg of p-nitrobenzaldehyde and 0.5 g of powdered, anhydrous magnesium sulfate are added and the solution is evaporated to 2 ml under reduced pressure. Then the solution is stirred 4 hr. At 25 ⁰ C. The obtained solution of O-trimethylsilyl-Np-nitrobenzylidene thienamycin trimethylsilyl is treated with 150 mg of dimethyl sulfate and the resulting mixture stirred for 1 hr. At 25 ⁰ C. The mixture is then poured into 10 ml of 0.1N pH 6 phosphate buffer, the mixture is stirred for 15 minutes and extracted with ether. The aqueous phase is chromatographed on 200 g of XAD-2 resin. A das is obtained by elution with water

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15835Y .. _?

Fraction containing N-methyl-thienamycin, which is evaporated and freeze-dried.

Example 1

Production of thienamycin benzyl ester

LEVEL A: N-Cp-Nitrobenzyloxycarbonyl ^ thienamycin sodium salt

43 mg of thienamycin are mixed with 10 ml of tetrahydrofuran / water (1: 1) ^{at 0 ° C.} 102 mg (10 equivalents) of sodium bicarbonate are added to the mixture while stirring rapidly, and then 4 equivalents of o-nitrobenzyl chloroformate are added dropwise over the course of 2 minutes while stirring. After 30 minutes, the pH is adjusted to 7 with 25% aqueous phosphoric acid and the solution is extracted three times with ether. The aqueous portion is ^{adjusted to a pH of 2.2 at 0} ° C. and mixed with 500 mg of solid NaCl. The cold, acidic solution is extracted three times with cold ethyl acetate. The combined extracts are quickly backwashed with cold sodium chloride solution, dried over magnesium sulfate and filtered. The filtrate is back-extracted with 10 ml of water to which 1.75 equivalents of solid NaHCO _{5 have been} incorporated. The extract is ^{freeze-dried at 20 °} C. in vacuo; this gives you the connection you want.

LEVEL B: N-Cp-Nitrobenzyloxycarbonyl-I-thienamycin-benzyl ester

The product from Step A) in 7.5 ml of ethyl acetate during 2 h. At 4 ⁰ C with an excess of phenyldiazomethane (4 ml of a solution containing 20 mg / ml ether) was reacted. The mixture is then evaporated at 20 ^° C. under reduced pressure to give a wet residue. The desired compound is determined by thin layer chromatography

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isolated on elution with ethyl acetate / ether (9: 1); you get 17 »5 mg of N- (p-nitrobenzyloxycarbonyl) -thienamycin-" benzyl ester.

LEVEL C: Thienamycin Benzyl Ester

The compound from stage B) is dissolved in ethanol, 35 mg of PtO _{0 are} added and the mixture is hydrogenated for 45 min

2
Pressure of 3 »51 kp / cm (50 lbs.). The reaction mixture is then centrifuged. The liquid is decanted and, after evaporation, is subjected to preparative thin-layer chromatography on silica gel, eluting with methanol / chloroform (1: 4)

Example 8

Production of Ν, Ν-dimethyl-thienamycin

A mixture of 18 mg of thienamycin benzyl ester, 14 mg of methyl iodide and 4 mg MgO in 2 ml of hexamethylphosphoramide is 1 hr, "stirred at 25 ⁰ C and then poured into hexane. The resulting precipitate of crude Ν, Ν-dimethyl-thienamacinbenzylester is in Added ethyl acetate and purified by thin layer chromatography on silica gel.

A solution of 10 mg Ν, Ν-dimethyl-thienamycin-benzyl ester in 1 ml of dioxane / water (4: 1) is for 2 hours. At a Hydrogenated pressure of 2.81 kp / cm (40 psi) in the presence of 10 mg palladium oxide. The catalyst is then filtered and the filtrate is shaken with a mixture of 5 ml of ethyl acetate and 5 ml of 0.01N pH 7 phosphate buffer. The watery one Phase is separated off, strongly concentrated and freeze-dried. This gives Ν, Ν-dimethyl-thienamycin.

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Example 9

N-benzyl- and Ν, Ν-dibenzyl-thienamycin

A mixture of 150 mg thienamycin, 300 ml benzyl bromide and 200 mg sodium bicarbonate in 10 ml 80% aqueous ethanol is ^{stirred at 23 °} C. for 5 hours. The resulting solution is evaporated to 2 ml under reduced pressure, diluted with 5 ml of water and extracted with ether. The aqueous layer is chromatographed on a column containing 100 ml of XAD-2 resin. The unreacted thienamycin is removed by elution with water. On elution with increasing concentrations of tetrahydrofuran, a fraction containing N-benzyl-thienamycin and then a fraction containing N, N-dibenzylthienamycin are recovered. The products are isolated by freeze drying.

Using the same method, but instead of benzyl bromide If the equivalent amount of allyl bromide is used, N-allyl- and Ν, Ν-diallyl-thienainycin are obtained.

Example 10

N-ethyl-N, N-bis-p-methoxybenzyl-thienamycin-p-methoxybenzyl ester iodide

A solution of 100 mg Ν, Ν-bis-p-methoxybenzyl-thienamycinp-methoxybenzyl ester and 0.5 ml of ethyl iodide in 1 ml of dimethylformamide is stirred 2 hrs. At 25 ⁰ C. The unreacted reaction components and the excess solvent are then evaporated off under reduced pressure and the residue is rubbed with ether. The solid product is obtained

- fiU -

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Example 11
N-ethyl-thi enamyc in

A solution of 120 mg of N-ethyl-l ^ lT-bis-p-methoxybenzylthienamycin-p-methoxyethyl ester iodide Rinse in 5 ml of 80% aqueous solution Ethanol containing 20 mg of sodium bicarbonate

is hydrogenated for 4 hr. at 25 ⁰ C and a pressure of 2.81 kp / cm (40 psig) in the presence of 100 mg palladium. The catalyst is then filtered off, the filtrate is evaporated down to 2 ml, it is diluted with 5 ml of water and extracted with ether. The aqueous phase is chromatographed on 100 ml of XAD-2 resin. On elution with water, N-ethyl-thienamycin is obtained, which is isolated by freeze-drying.

Example 12

Ν, Ν-dimethyl-N-benzyl-thienamycin

A solution of 200 mg of N-benzyl-thienamycin in 5 ml of 50% aqueous dioxane is adjusted to a pH of 8.4 (titrated). Then add over 10 min. With stirring Add 0.5 ml of dimethyl sulfate in 2 ml of dioxane. The pH value is set by adding 1N sodium hydroxide solution using an automatic Titration device held at 8.4. The mixture is then stirred for an additional hour, then with Diluted 10 ml of water, adjusted to pH 7 and extracted with ether. The aqueous phase is increased to 5 ml concentrated and chromatographed on 200 ml of XAD-2 resin. The column is eluted with water and then with 20% aqueous tetrahydrofuran. The N, N-dimethyl-ir-benzylthienamycin is isolated from the tetrahydrofuran eluate by freeze-drying.

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Example 13
N, .N-dimethyl-thi enamyc in

Ν, Ν-Dimethyl-N-benzyl-thienamycin comes in the form of a solution hydrogenated according to Example 11; this gives N, N-dimethylthi enamyc in.

Example 14
N-salicylidene thienamycin benzyl ester

115 mg of thienamycin are dissolved in 4 ml of 50% aqueous dioxane. The solution is ^{cooled to 0} ° C. and adjusted to a pH of 5 with 1N sulfuric acid (titrated). Then, while stirring vigorously, 60 mg of phenyldiazomethane in 0.9 ml of dioxane are added over the course of 5 minutes, the pH being kept at 5 to 5.5 while monitoring with a pH controller. After a further 5 minutes of reaction, the mixture is extracted with ether. The aqueous layer is adjusted to pH 8.3 with sodium bicarbonate solution and extracted with ethyl acetate. The solution containing thienamycin benzyl ester is then mixed with 35 μl salicylaldehyde and anhydrous magnesium sulfate. The solution is concentrated to 1 ml on a rotary evaporator and left to stand ^{at 25 ° C. for 1 hour.} The course of the reaction is followed by thin-layer chromatography on silica gel in 20% methanol / chloroform. A new spot appears at R "0.8. The product is isolated by preparative thin-layer chromatography in 50 io ethyl acetate / chloroform and appears as a yellow band at Ή. _Ψ 0.32; UV) 259 πιμ and 322 πιμ with the same intensity; d (CH ₃ CHOH); 6 * 7 to 7 and 5.7 to 6.4 (multiplets); 4.72, s, (OCH ₂ ); 2.5 to 7.5

(Multiplets, aromatic H) and 1.75, S (C = N).

709822/10 29 ~ ⁶⁶ ~

The same method, but using diphenyldiazomethane instead of phenyldiazomethane, gives salicylidene-thienamycin-benzhydryl ester;

NMR 8.67 d (CH ₃ COH); 5.65 to 7.1 (aliphatic multiplets);

2.5 to 3.2 (aromatic multiplets), 1.67 (C = N).

Furthermore, using the same method, but using p-nitrobenzaldehyde instead of salicylaldehyde, p-NitiObenzyliden-thienamycin-benzyl ester and the corresponding Benzhydryl esters. Thin-layer chromatography using CHCl, / ethanol (5: 1) gives a R-p value of 0.8. In an analogous manner, one obtains using of benzaldehyde, p-bromobenzaldehyde, p-dimethylaminobenzaldehyde, 5 »5-dimethyl-1,3-cyclohexanedione, dimethylaminoacetaldehyde or isobutyraldehyde instead of salicylaldehyde, the corresponding benzhydryl and benzyl esters.

Example 15

N-ethyl-Np-bromobenzylidene-thienamycin-benzyl ester-ethyl sulfate .

200 mg of p-bromobenzylidene-thienamycin-benzyl ester are dissolved in 2 ml of methylene dichloride, and 0.2 ml of diethyl sulfate is added to the solution. Then the solution is stirred for 5 h. At 40 ⁰ C, adding 20 ml ether and the precipitated N-ethyl-Np-bromobenzylidene thienamycin benzyl ester from ethosulphate.

Example 16

N-ethyl-thienamycin

A solution of 200 mg of N-ethyl-N-p-bromobenzylidene-thienamine-

709822/1029 ~ ^6? "

ethyl sulfate in f ml of 80% ethanol with a content of 40 mg Sodium bicarbonate is blown for 4 hours at 2.81 kgf / cm (40 psig) in the presence of 0.2 g of palladium oxide hydrogenated. The catalyst is then filtered off. The filtrate is evaporated to 2 ml and diluted with 5 ml of water and extracted with ether. The aqueous phase is chromatographed on 200 ml of XAD-2 resin. Elute the column with water. The fractions containing the N-ethyl-thienamycin are freeze-dried.

Example- 17
N-methyl-thienamycin benzyl ester

A solution of 50 mg of ET-methyl-thienamycin in 1 ml of water and 1 ml of dioxane is ^{cooled to 0} ° C. and adjusted to a pH of 5 with 1N sulfuric acid. 37 mg of phenyldiazomethane in 0.5 ml of dioxane are then added over a period of 5 minutes, the pH value being maintained at 5-5-5 with the aid of an automatic titration device. The mixture is then diluted with 5 inl V / water and extracted with ether. The aqueous phase is covered with a layer of ethyl acetate, cooled and adjusted to a pH of 2.5. The ethyl acetate phase is centrifuged off, the aqueous phase is adjusted to pH 8.0 with sodium bicarbonate and extracted twice with ethyl acetate. The extracts are combined and evaporated. The product is isolated by preparative thin-layer chromatography on silica gel using chloroform / methanol (5: 1) as the solvent.

Using the same method, however, from N, N-dimethylthienamycin goes out, one receives Ν, Η-Dimethyl-thienamycinbenzylester.

709822/102 9

- 68 -

Example 18

N-o-Hydroxyl enzyl-th.ien.amyc in-b enzyl e s t er

40 mg of N-salicylidene-thienamycin benzyl ester in the form of a solution in 1 ml of dioxane for 2 hours. "Reported at 23 ⁰ C and a pressure of 2.81 kp / cm (40 psig) was hydrogenated in the presence of 10 mg of platinum oxide. Then The catalyst is filtered off and the filtrate is evaporated in. The residue is taken up in chloroform and chromatographed on silica gel, essentially pure n-hydroxybenzyl-thienamycin-benzyl ester being obtained.

Using the same method »but instead of N-salicylidene-thienamycin-benzyl ester in each case the equivalent Amount of N-benzylidene-thienamycin-benzyl ester, M-p-bromobenzylidene-thienamycin-benzyl ester, N-dimethylaminobenzylidene thienamycin benzyl ester, N-dimethylaminoethylidenthienamycin benzyl ester or N-2-methylpropylidene-thienamycin-benzyl ester uses, one receives N-benzyl-thienamycin-benzyl ester, N-p-bromobenzyl-thienamycin-benzyl ester, N-p-dimethylaminobenzyl-thienamycin-benzyl ester, N-dimethylaminoethyl-thienamycin-benzyl ester or N-isobutylthienamycin benzyl ester.

Example 19

N, N-trimethyl-thienamycin

A solution of 150 mg thienamycin in 7 »5 ml 0.1N pH 7 phosphate buffer and 7 »5 ml of dioxane is brought to a pH of 8.4 adjusted, treated with 1 ml of dimethyl sulfate and stirred rapidly for 40 minutes, the pH being adjusted by adding 1N sodium hydroxide solution is kept at 8.4. The solution is then extracted twice with ether. The aqueous phase is up to

709822/102 & -69-

4 ml evaporated and onto a 200 ml XAD-2 resin containing Pillar abandoned. The column is eluted with water and the course of the separation is followed by high-pressure liquid

1 row
Chromatography on C -Bondapak resin using tO ^ strength aqueous acetonitrile. The product has 1.2 times the retention time of thienamycin. The fractions containing the product are combined, evaporated and freeze-dried. 41 mg of Ν, Ν, Ν-trimethyl-thienamycin are obtained. The electrophoresis (50 V / cm, 2 hours, pH 7 buffer) results in a bioactive zone which migrates by 6 cm in the direction of the cathode. The KMR spectrum (60 MHz, DpO) shows a strong methyl singlet at 6.78? with an integral 3X side chain methyl doublet at 8.7 ?; UVA _ma x ² 9 & ^m V * * # 185.

Using the above method, but using 0.25 ml of dimethyl sulfate begins and the reaction is allowed to proceed for 15 minutes, a mixture is obtained which contains N-methyl- and N, N-dimethylthienamycin and is separated by column chromatography.

Example 20

Production of Ν, ΪΓ, Ιί-trimethyl-thienamycin

0.2 ml of hexamethyldisilizane and 0.1 ml of trimethylchlorosilane are added to a suspension of 20 mg of Ν, Ν-dimethylthienamycin in 10 ml of tetrahydrofuran in a nitrogen atmosphere while stirring. The mixture is ^{stirred vigorously at 23 °} C. for 20 minutes and then centrifuged. The supernatant solution is evaporated under reduced pressure. The remaining oil is dissolved in 1 ml of tetrahydrofuran, 0.05 ml of methyl iodide is added with vigorous stirring and the mixture is stirred for 30 minutes. Then 5 ml of ethyl acetate and 5 ml of 0.1N pH 4 phosphate buffer are added and the mixture is stirred for a further 15 minutes

709822/1029 " ⁷ °"

25 ⁰ C and then adjusts it to a pH of 7. The aqueous layer is then separated off, concentrated to 1 ml and applied to a column containing 20 ml of XAD-2 resin. It is eluted with water and then with 10 tetrahydrofuran. A fraction containing Ν, Ν, Ν-trimethyl-thienamycin is obtained which, on freeze-drying, gives a solid product.

Example 21
^ N-Dimethyl-thienamycin-pivaloyloxymethyl ester

A solution of 30 mg Ν, Ν-dimethyl thienamycin and 25 mg of pivaloyloxymethyl bromide in 0.2 ml of hexamethylphosphoramide is stirred for 1 hr. At 23 ⁰ C. 5 ml of ethyl acetate are then added and the mixture is extracted successively with aqueous sodium bicarbonate solution, water and saturated sodium chloride solution. The organic phase is dried, concentrated strongly and chromatographed on a 20.32 χ 45 »72 cm (8 χ 18 in.) 1000 μ-SiOp plate using chloroform / methanol (5: 1) as the solvent. The strip containing the N, N-dimethyl-thienamycin-pivaloyloxymethyl ester is scraped off and eluted with ethyl acetate.

Example 22

NjN-dimethyl-thienamycin ^ -methyl ^ -butenylester-hydrochloride

A solution of 30 mg of N, N-dimethylthienamycin in 0.5 ml of 3-methyl-2-butenyl alcohol, which contains 3 »6 mg of HCl, is mixed with 21 mg of dicyclohexylcarbodiimide and ^{stirred at 23 °} C. for 1 hour. The dicyclohexylurea is then filtered off, the piltrate is evaporated and the residue is rubbed

709822/1029 ~ ^{? 1} ~

15833Y

with ether on. A solid material remains, which is N ^ -dimethyl-thienamycin ^ -methyl ^ -butenyl ester hydrochloride contains.

Using the above method, but using methylthioethanol if 2-methyl-2-butenol is used instead of 2-methyl-2-butenol, N, N-dimethyl-thienamycin methylthioethyl ester is obtained.

Example 23

0-Acetyl-N, N-dimethyl-thi enamyc in

100 mg of Ν, Ν-Mmethyl-thienamycin are added to a mixture of 0.3 ml of acetic anhydride and 1 ml of pyridine. The mixture is allowed to ^{react for 3 hours at 23 °} C. and then evaporated to dryness in vacuo. The solid residue is dissolved in water and the solution is chromatographed on 100 ml of XAD-2 resin. After elution with water, the product is eluted with 10% tetrahydrofuran. The fractions containing the O-acetyl-N, N-dimethylthienamycin are combined, evaporated and freeze-dried.

Example 24 Thienamycin benzyl ester

A solution of 47 mg of thienamycin in 1 ml of water and 1 ml of dioxane is ^{cooled to 0} ° C. and adjusted to a pH of 5 with 1N sulfuric acid. 37.2 mg of phenyldiazomethane in 0.5 ml of dioxane are then added over the course of 2 minutes, the pH being kept at 5 with the aid of an automatic titration device. After a further 5 minutes, 5 ml of water are added and the mixture is extracted with ether. The aqueous phase is covered with a layer of ethyl acetate, cooled and then on

709822/102 9

adjusted to a pH of 2.5. The ethyl acetate phase is separated off, the aqueous phase is adjusted to pH 8 with sodium bicarbonate and extracted twice with ethyl acetate r The two extracts are combined and dried over anhydrous magnesium sulfate. Thin layer chromatography on silica gel using methanol / chloroform (1: 5) gives a single ninhydrin positive spot at R- 0.24. The UY spectrum of the ethyl acetate solution shows a maximum () J at 318 πιμ with an optical density from 250.

Example 25

ff ^ I-Dimethyl-O-sulfo-thienamycin-benzyl ester Man. a solution of 39 mg of N, N-dimethylthienainycinbanzyl ester in 0.3 ml of pyridine is mixed with 17 mg of sulfur trioxide / pyridine. The mixture is stirred at 25 ° C. for 3 hours. The excess pyridine is then evaporated off under reduced pressure, the residue is taken up in 5 ml of water, into which 10 mg of sodium bicarbonate have been incorporated, and extracted once with ethyl acetate. The aqueous solution is concentrated to 2 ml and chromatographed on 50 g of XA.D-2 resin. The fractions containing the Ν, ΙΤ-dimethyl-O-sulfo-thienamycin-benzyl ester are combined, evaporated and freeze-dried.

7Q9822 / 1Q29

- 73 -

^15853Ϊ ^. 2652576

Example 26
! ", IT-dimethyl- ^ O-sulfo-thienamycin sodium salt

24 mg NfN-dimethyl-O-sulfo-thienaniyein-benzyl ester in the form of a solution in 1 ml V / water with a content of 5 mg sodium bicarbonate are for 2 hours at 23 ⁰ C and a pressure of 1 atm in the presence of 20 mg of palladium oxide hydrogenated. The catalyst is then filtered off and the filtrate is chromatographed on 20 g of XAD-2 resin. The fractions containing the H> N-dimethyl-0-sulfo-thienamycin-1 · sodium salt are combined, evaporated and freeze-dried.

Example 27

O-formyl-N, H-dimethyl-thienamycin-benzyl ester

A solution of 100 mg NiN dimethyl thienamycin benayl ester in 1 ml pyridine is mixed with a mixture of 100 mg formic acid and 200 mg acetic anhydride. The resulting mixture is stirred for 2 h at 25 ⁰ C and then removed under reduced pressure of the excess reagents, the residue is nimrit up in ethyl acetate and the product is isolated by Dümischichtchroraatographie on silica gel using ethyl acetate / chloroform. (1: 1) as Solvent.

Example 28

0-formyl -Ή , Η-dimethyl -t hienamyci n

50 mg of O-formyl-HtN thienamycin benzyl ester in the form of a solution in 2 ml of 90 $ strength of ethanol for 4 hr. At 23 ⁰ C

709822/1029

15833Y

and a pressure of 1 atm in the presence of 50 mg of 1Obiger Palladium-activated carbon hydrogenated. The catalyst is then filtered off, the filtrate is evaporated and the the residue containing O-iOrmyl-UjlT-dimethyl-thienamycin chromatographically on XAD-2 resin.

Example 29

Production of N-thioformyl-thienamycin

raw
S.

Th- -NH-CH

A stoppered flask is placed under positive nitrogen pressure silylated thienamycin [Th (TMS ^), obtained from 100 mg of thienamycin according to Example 4-3 dissolved in 9 ml of dichloromethane. The magnetically stirred solution is added a solution of 60 μΐ triethylamine in 1 ml of dichloromethane and then with 100 μΐ ethyl thioate. Do The reaction solution is quickly added to a solution of a 0.1N pH 4 phosphate buffer (20 ml) for 1 hour while stirring. The mixture is stirred for 5 minutes and then made up with 1N NaOH to a pH of 7 »0. The aqueous phase is separated off, washed twice with 20 ml of ethyl acetate each time and chilled in an ice bath. Then the solution is coated with 15 ml of ethyl acetate and the pH of the mixture is adjusted Stir with 1N phosphoric acid to 3 »5. The organic phase is separated off and the buffered aqueous solution is washed twice with 15 ml of ethyl acetate each time. The combined ethyl acetate washes are concentrated to half the volume and sublayer et with 10 ml of water. Then you set the pH of the mixture with solid sodium bicarbonate to 7 »0

. - 75 709822/102 9

15833Y

a. The aqueous phase is separated off and freeze-dried! whereby the sodium al ζ of N-thioformyl-t-menamyoin is obtained.

Example 30

i ~ OH

· —COOH
N- (thiobenzoyl) thienamycin

A solution of 55.8 mg (0.2 mmol) of thienamycin in 18 ml of dioxane / water (1: 1) is treated with 263 mg (3.1 mmol) of sodium bicarbonate and cooled to 0 ⁰ C. Two 200 μl aliquots of a solution of 100 mg of thiobenzoyl chloride in 0.6 ml of dry dioxane are then added to the vigorously stirred solution at 15 minute intervals. Each aliquot of solvent contains 0.2 mmoles of thiobenzoyl chloride. 15 minutes after the second addition, the reaction solution is washed twice with 8 ml of ether each time, 8 ml of ethyl acetate are added to the aqueous phase and, while stirring rapidly at ⁰ ° C., it is brought to a pH of 2 with 20% phosphoric acid, 2 a. The layers are separated and the aqueous layer is washed with 3 ml of ethyl acetate. The combined ethyl acetate layers are dried over magnesium sulfate. After the desiccant has been separated off, 10 ml of water are added to the ethyl acetate solution and the product is extracted into the aqueous phase by adding 50 mg (0.62 mmol) of sodium bicarbonate (pH 7.4) ^{at 0 ° C. with stirring.} The layers are again separated from one another, an aqueous phase containing the N- (thiobenzoyl) thienamycin sodium salt being obtained which is freeze-dried; IiMR in D ₉ O: cT 7 »3 to 7.9» characteristic of the

7Q9822 / 1029 - 76 "-

15833Y

Example 31

Production of N-methyl-thienaraycin (I) and E-benzylthienamycin (II)

A mixture of 100 mg Έ- (thioformyl) -thienamycin and 0.5 g of neutral Raney-nickel in 100 ml of 90 $ aqueous ethanol is stirred for 2 hr. At 23 ⁰ C, inschließend is filtered from the nickel and chromatograph ert! The Piltrate on a column (100 ml) with Dowex 50x4 resin (Na ⁺ form; 37 / U · or 400 mesh). The column is eluted with water. The fractions containing the N-methyl-thienamycin are combined and freeze-dried.

J Same method, but instead of H- (lhioformyl) -thienamycin the equivalent amount of "- (! Ehiobenzoyl) -thienamycin uses, one gets.IT-Benzyl-thienamycin.

70982 2/1029

- 77 -

1-5833Y '

Example 32

Subject to those in the preceding examples and in the description The following compounds according to the invention are produced in the methods explained:

> 3

5 6 7 C ^)

SCH ₂ CH ₂ NR RR -A ^w

COXR

X E ³ & r5 E ⁶ R? A. Ver
am
manure 0 H R. CH ₃ CH ₃ CH ₃ Cl;
I. .1) 0 CH ₃ -CH ₀ -OC-CH-,
Z ti O
0 CH ₃ CH ₃ CH ₃ - ^: 2) 0 SO ₃ H - CH ₃ CH ₃ CH ₃ - ■ 3) 0 H N / A CH ₃ CH ₃ ^C 2 ^H 5 - 4) 0 H - ^C 2 ^H 5 CH ₃ - 5) 0 H H ^C 2 ^H 5 C ₂ H ₅ - - 6) 0 H H , "CH ₂ CH = CH ₂ H H Cl 7) 0 H -CH ₂ -O-CH ₂ CtCH ₃ ). "" -Cxi ^ C H «CIH ο H - - 8th) 0 H H -CH0 ₂ H - - 9) 0 H H -CH02 CH ₃ - - 10) 0 H H -C-0 3 CH ₃ - - - 11) 0 H H CH0 ₂ CH ₃ CH ₃ - 12) 0 H - -CH ₂ -CH = CH ₂ H H Cl 13) 0 H -CH ₂ -CH = CH ₂ OTT _O OtT
L-Ii _n V, -UrI _n
Zt Z H H Cl 14) -CH ₂ -C = CH ₂ CH ₃ CH ₃ -

7 0 9 3 2 2/1029 - 78 -

15833Y

Ver -τ. 5 6 am- χ R ^ R Ε? R ^D R 'A dung

15) OH -CH ₂ CH ₂ N (C ₂ H ₅ I ₂ CH ₃ HHH SO

16) 0 -CH ₂ OCH ₃ p-Hitrobenzyl -CH ₂ OCH ₃ -CH ₂ OCH ₃ -

-CH ₂ CH ₂ CH ₃ -CH ₂ CH = CHCH ₃ HH Cl H -CH ₂ CH ₂ CH = CH H - -

. NH

TT "^ f ^ tl II TJ

ο Ii ^. ^ 2 ^. ^ H

17) S. H 18) 0 H 19) 0 H 20) 0 -COCH,

21) 0 H H

£ ■ έ. Ζ / L

22) 0 H -CH ₂ CH ₂ N- (CH ₃ J ₂ - -CH ₂ CH ₂ -N- (CH ₃ J ₃ H H-HPO ₄

23) 0 HH -CH ₀ -C-CH ₇ H

* 11 -3

24) 0 HH -CH ₂ CH ₂ (OCH ₃ J ₂ H

25) 0 HH -CH ₂ -C = CH H

26) 0 HH -CH ₂ CH ₂ O-CH ₂ CH ₂ -H

27) 0 H -CH-C-0 -CH-C-0 CH,

i « ί Il J

0 0

28) 0 H CH ₂ S-CH ₃ -CH ₂ SCH ₃ -CH ₂ SCH ₃ -

29) 0 H -CH ₀ -OC (CH-,), -CH _-3 -OC (CH,), H.

ο ο

30) 0 Si (CH ₃ J ₃ Si (CH ₃ J ₃ -CH ₂ CH ₂ C ^ n H

31) OHH-CH ₉ CH ₉ -C-OCH, H.

* O ^J

709822/1029

- 79 -

2 ö b 7 b 7 b 15833Y

Connection ^ 5 6 χ r ^j RR ~ __R R

32). OHH _ <f ~ \ _NO ₂

33) O -C-CH ₂ NH ₂ H ~ ^C 2 ^H 5 ~ ^C 2 ^H 5 ^H CH ₃ COO O

CH

34) OHH -CH ₉ - / j

^V CH

39) O H 40) O H 41) O H 42) O H 43) O H 44,) O H 45) O H

46) O H

-CH.

35) O H H -CH (CH ₃ ) ₂ H 36) O H H -CH ₂ CH ₂ CH ₂ CH ₃ H 37.). O H H -CH ₉ CH-CH ₀
λ ι 3 H CH ₃ 38) O H H -CH CH ₂ CH ₃ H CH.

H -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ H

H -CH-CH ₂ CH ₂ CH ₃ H CH ₃

H -CH ₂ CH-CH ₂ CH ₃ H CH ₃

H -CH ₂ CH ₂ CH-CH ₃ H CH ₃

H -CH-CH-CH ₃ H CH ₃ CH ₃

H -CH ₂ -C (CH ₃ J ₃ H

H -CHCH ₂ CH (CH ₃ ) ₂ H CH,

47) OHH -CH ₂ CH = CH ₂ H

48) OEH -CH ₂ CH-CH ₃ H

7 0 9 8 2 2/1029 ^CH 3

- 80 -

2652678

Connect
manure X R ⁵ R. R ⁵ -ο R ⁶ R ⁷ A 49) 0 H H -CH-CH = CH ₂ -OH ₂ OH ₂ -Q H - - CH ₂ 50) 0 H H -CHpCH = CH-CH ^ -CH ₂ -Q TT- - 51) 0 H H -CH ₂ -CH ₂ -CH = CH ₂ -CH ₂ -CH = /] H 52) 0 H H -CH-CH = CHp H 53) 0 H H -CH ₂ -CH = CH-CH ₂ CH ₅ ίΓΊΊ H 54) 0 H H —C Hp C H = C H-C Hp C Hp C Η ·? H - - 55) 0 H H -CH ₂ -C = CH-CH ₂ CH ₃ H OH ₃ 56) 0 H H -CH ₂ -CH ₂ -CH = CH-CH ₅ H 57) 0 H H ' CH ₂ CH = CH-CH-CH ₅ H 58) 0 H H CH ₅ H 59) 0 H H H 60) 0 H H H- - 61) 0 H H H - - 62) 0 H H H- 63) 0 H H H 64) 0 H H H 65) 0 H H H

66) 0 H H _ (j fl H

709822/10 2981 -

158331

Link

X R-

R ⁷ A

67)

H H

68) ο H H 69) 0 H H 70) 0 H H 71) 0 H H 72) 0 H H 73) τι TT

74) 75)

76)

77)

O H

O H

O H

0 H

-CH ₂ -CH = CH-0 OCH-

-CH

-CH-

-CH.

-CH-, -CH ₀ CH = CH.

-CH -CH

-CH

-CH

-CH ₂ CH = CH-CH ₃

-CH-CH = CH-ι *

CH,

-CH = CH-CH ₂ CH ₃

-CH ₇ -C = CH ₂ -CH ₃ CH ₃

, -CH ₉ -CH = CH-CH-CH, ■ j -, j

-CH

78)

H H -CH

3 CH ₀ -J

79)

O H

-CH ₃ -CH ₂ -CH = CH-0

0 = phenyl

7 0 9 8 2 2/1029 - 82 -

15833Y

B ice ρ i e 1 33

λοο

Manufacture of medicines

Such a single dosage form is obtained by adding 120 mg Ν, ίί, Τί-trimethyl-thienainycin-chloride with 20 mg of milk sugar and 5 mg Magnesium stearate mixed and the mixture (145 mg) poured into a # 3 gelatin capsule. Analog can be done using a higher Active ingredient and a smaller amount of lactose in other preparations

ITr.3 can be entered in gelatin capsules. If necessary is to mix more than 145 mg of the components together » You can also get larger capsules as well as compressed tablets and pills produce. The following examples are intended to explain the manufacture of pharmaceuticals:

Tablet formulation

N, N, N-trimethyl-thi enamycin chloride

Corn starch, U.S.P. Dicalcium phosphate Milk sugar, U.S.P.

mg per tablet

125 6

192 190

The active ingredient is mixed with the dicalcium phosphate, the milk sugar and about half of the corn starch. The mixture is granulated with 15 ^ 6mg sodium Maize starch paste and coarsely sieved out the granules, dried at 45 ⁰ C and again sieved through sieves ITr.16. The remainder of the corn starch and magnesium stearate are then added and the mixture is compressed into tablets each about 0.5 in. In diameter and 800 mg in weight.

709822/1029

- 83 -

15Θ33Τ

AO λ

Parenteral solution

ampoule

N, N, N-T rime thyl-thienamyc in-

chloride 500 mg

Diluent: sterile water for Injections 2 ml

Eye solution

N, N, N-trimethyl-thienamycin-

chloride 100 mg

Hydroxypropylmethylcellulose ^ν · "5

sterile water 4.8. to 1 ml

Ear solution

N, N, N-trimethyl-thienamycin-

chloride 100 mg

Benzalkonium chloride 0.1 mg

sterile water qs to 1 ml

Local ointment

N, N, N-trimethyl-thienamycin-

chloride 100 mg

Polyethylene glycol 4000 U.S.P. 4OO mg

Polyethylene glycol 400 U.S.P. 1.0 g

The active ingredient of the above formulations can be administered alone or in combination with other biologically active substances, for example other antibacterial agents such as lincomycin, a penicillin, streptomycin, novobiocin, gentamicin, neomycin, colistin and kanamycin, or with other therapeutic agents such as probing oath .

709822/1029

Manufacture of alternative raw materials

In addition to thienamycin, it is evident to the person skilled in the art that its various isomers can be used alone or as mixtures as starting materials in the preparation of the compounds according to the invention. Some of these isomers are available from natural products of fermentation (see, below). However, by means of total synthesis, all isomers can be obtained (see below) in the form of a mixture of H diastereoisomers which have bactericidal activity and which can be separated by customary technical processes. The H diastereoisomers (2 cis, 2 trans) can be separated by chromatography. The separation of a given d / 1 pair with optically active acids or bases takes place according to conventional methods. It should be noted that the absolute configuration of the first identified starting material (I) is 5R 6S 8R.

- 85 709822/102 9

Production of thienamycin by total synthesis

OH

J_J

COOH

Level A:

Preparation of 4- (2-acetoxyvinyl) azetidin-2-one

H ₀ C = CH-CH = CHOC-CH. + O = C = N-SO ₀ Cl

J- ti -J J-

CH = CHOCCH,

.CH = CHOCCH-

SO ₂ Cl

-NH

A solution of 1.0 ml of distilled chlorosulfonyl isocyanate (1.65 g> 11.7 mmol) in 2.5 ml of anhydrous diethyl ether is cooled under Np in a bath of -20 C.

A solution of 2.5 g of L-acetoxybutadiene (22 mmoles) in 2.5 ml anhydrous ether is similarly cooled under nitrogen in a -20 ⁰ C bath.

The chlorosulfonyl isocyanate solution is added dropwise to the Acetoxybutadiene solution added by means of a polytetrafluoroethylene tube immersed in the CSI solution and

709822/10 29

- 86 -

is pressurized with N _2. The addition takes 10 minutes. Fenig or no color is seen and the reaction is stirred for 0.5 hours at -20 ⁰ C. The solution is clear and light yellow in color.

A solution of 2 g of sodium sulfite and 5 g of K ₂ HPOn in 20 ml of H ₃ O is prepared during the aforementioned 0.5 hour reaction time and cooled on an ice bath; 20 ml of ether are added to the mixture with vigorous stirring on an ice bath. At the end of the 30-minute reaction time, the reaction mixture, again using Np pressure and a polytetrafluoroethylene tube, is transferred from the reaction flask, which is kept in a bath at -20 ° C., to the vigorously stirred hydrolysis mixture. The rapid, dropwise addition is complete after 5 minutes. The hydrolysis is allowed to proceed for an additional 5 minutes. The hydrolysis mixture has a pH of 6-8, preferably pH 8.

The phases are separated, with a yellow-orange resin remaining in the aqueous phase. The ether phase is dried directly over MgSO ₁ . The aqueous / resin phase is extracted three times with 50 ml portions of ether, each portion being added to the starting ether / KgSOr mixture.

The dried extracts are filtered and concentrated to 5 ml under a stream of Np; part of the product -is crystallizes at this stage.

A column of 10 g of Baker's Kiselgel packed in Ether, here and the ether concentrate is given on top and allowed to flow through. The residue contained in the flask is rinsed three times with 2 ml of ether, each time pipetted off and applied to the column. Then it is eluted with ether.

. - 87 -709822/1029

The first 25 ml are mostly empty volumes. The next 5 10 ml fractions are collected, followed by 3 50 nl fractions and all are reduced in volume under a stream of Np. The product crystallizes from fractions H to 6 with traces in 3 and 7- Fractions 1 to 3 contain a yellowish, pungent-smelling material which becomes resinous on standing. Yield: 100 mg as a mixture of the cis and trans isomers.

Level B;

Production of * l- (2-acetoxyethyl) -2-acetidinone

, OCCH ₃

A solution of ⁱ i- (2-Acetoxyvinyl) -2-azetidinone (10.0 g, 0.065 mol) in 200 ml of ethyl acetate containing 100 mg of 10% palladium / C catalyst is hydrogenated in a Parr bulk, ler hydrogenated at 25 C under a pressure of 2.8 kg / cm in the course of 15 minutes. The mixture is filtered through a bed of Supercel and washed with additional ethyl acetate. The combined filtrate is washed in vacuo, M- (2-acetoxyethyl) -2-azetidinone (10.0 g) being obtained as a crystalline solid. When recrystallizing from ether, white crystals are obtained: F. HH to H7 ° C; IR (CHCl ₃ ) 5.66, 5.7 **; KMR (CDCl ₃ ) T3, Hk

(broad s, 1, NH), 5.82 (m, 2, CH ₀ OCOCH-), 6.29 (m,

- * - .0

1, C-HE, 6.87 (1/2 AB pattern continues to be quadruple

709822/1029

divided by C-4H and NH, 1, J = 12.8Hz,

* * gladly

J = 4.5 HH _NH = 1.9 Hz, 7.38 (1/2 AB pattern is still divided four times by C-4H and NH, 1, J _em = 12.8 Hz, J = 2.3 Hz, J _NH = 1.0 Hz). 7.93 and 8.02 (s an m, total 5, OCOCH, and CH _n CH _n OCOCH,., Respectively).

Level C:

Production of 4- (2-hydroxyethyl) -2-azetidinone

Under nitrogen at 0 ⁰ C a solution of 4- (2-acetoxyethyl) -2-azetidinone (2.24 g, 0.014 mol) in 25 ml of anhydrous methanol (77 mg, 1.4 mmol) with a solution of sodium methylate in Treated 5 ml of anhydrous methanol. After stirring for one hour, the solution is neutralized with glacial acetic acid. Removal of the methanol in vacuo gives the crude 4- (2-hydroxyethyl) -2-azetidinone as an oil. The product is purified by silica gel chromatography , eluting with 10% methanol / CHCl 2, to give 1.55 g of the alcohol: mp 50 ° C; IR (CHCl ₃ ) JU 5.67; KMR (CDCI ₃ ) - ^ 3.20 (broad s, 1, NH), 6.24 and 6.28 (m at t, total 3, C-4H and CH ₂ OH respectively), 6.90 (broad s an 1/2 AB pattern further splits into four by C-4H and NH, total 2, OH and C-3H accordingly, ^J gem ^{= 13} · ^0Ηζ » ^J vic = MHz, J ^ = 1.6Hz), 7, 42 (1/2 AB pattern further splits into four by C-4H and NH, 1, C-3H, ^J gem ^{= 1} ^ ⁰¹¹ Z * J _vic = 2.2Hz, J _NH = 1.1Hz), 816 ( m, 2, CH ₂

-Rq-

709822/102 9 ^y

Level D-:

Preparation of 8-oxo-2,2-dimethyl-3-oxa-1-azabicyclo- / 3,2.07-octane

OH

-NH

A solution of ⁱ t- (2-hydroxyethyl) -2-azetidinone (1.87 g, 0.016 mol) and 2,2-dimethoxypropane (1.69 g, 0.016 mol) in 25 ml of anhydrous methylene chloride is treated with boron trifluoride etherate (0201 ml , 0.002 mol) at 25 ° C. The resulting solution is stirred for 10 minutes. Removal of the solvent under reduced pressure gives an oil (2.5 g). Chromatography of the crude product on a silica gel column using 2: 1 ethyl acetate / benzene as the eluent gives 8-oxo-2,2-dimethyl-3 -oxa-1-azabicycl3yii, 2.07-octane (1.59 g) as a crystalline solid. Recrystallization from ether / hexane gives a product with a melting point of 60 to 6l C. IR (CHClJp: 5.73 (ß-lactam)

KMR (CDC1 ₃ ) 7: 6.02-6.28, m, 2H, C-4 methylene 6 ₅ 22-6.62, m, IH, C-6 methine 6.90, dd, IH, J _{7 7} = llJHz, J ^ _v = ^, 5 Hz C-7 proton cis to C-6H 7, ^ 7, dd, IH, J _{7 7} = 14 Hz, Jr _v = C-7 proton trans to D-6H 7, 82 ~ 8.68, m, 2H, C-5 methylene 8.23, s, 3H | _c _ _{2 methyls} 8.57, s, 3Hj

709822/1029 " ⁹ °"

408

Level E:

Production of 8-oxo-2,2-dimethyl-7 ^clC - and ß- (1-hydroxy ethyI) - (3-oxa-1-azabicyclo - /%, 2.07-octane

To a solution of 1.1 equivalents of freshly prepared lithium diisopropylamide in anhydrous tetrahydrofuran under a stickst off atmosphere at -78 ⁰ C a solution liird of 8-oxo-2,2-dimethyl-3-oxa-l-azabicyclo / 5, 2, o7-octane in anhydrous tetrahydrofuran, which has been cooled to -78 ° C., was added. After 2 minutes, the lithium enolate obtained is treated with excess acetaldehyde. The solution is stirred for 30 minutes at -78 ⁰ C and then poured into water. The aqueous phase is saturated with sodium chloride and extracted with ethyl acetate. The combined ethyl acetate solutions are dried over magnesium sulfate and filtered. The filtrate is evaporated under reduced pressure to give the crude product. Purification by means of chromatography over silica gel with the blending of ethyl acetate / benzene gives 8-oxo-2,2-dimethyl ^ ci'- and β- (l-hydroxyethyl) ~ 3-oxa-l-azabicyclo- / 3, 2, o7-octane.

Data for 8-oxo-2,2-dimethyl-7ß- (l-hydroxyethyl) -3-oxal-azabicyclo-A, 2, o7-octane:

IR (CH ₂ Cl ₂ ) ^ i: 5.72p. (ß-lactam)

5.53-6, ^ 3, m, ^ H, C-H methylene +

C-6 methine + C-9 methine

KMR (CDCl ₃ ) T '

709822/1029

- 91 -

December 28, 1976 15833Y

6.90, dd on wide s, 2H, ^J ₇ ο =

J _{6 7} = 5.5 Hz, C-7 methine + OH

7.70-8.83, m, 2H, C-5 methylene

8.27, s, 3H)

8.60, s, 3h] ^C - ^{2 meth} ^

8.78, d, 3H, J _{9 10} = 6.5 Hz, C-IO methyl

Data for 8-oxo-2,2-dimethyl-7c ( ^: - (l-hydroxyethyl) -3-oxa-lazabicyclo- / 4,2.07-octane:
IR (CHCl ₃ ) p: 2.9 broad O-H

5.73 β-lactam

KMR (acetone - d-Oe ^: 4.23-3.33, m, C-9 methine + C-4

Methylene + C-6 methine

3.33, broad s, OH

2.83, dd, J = 2Hz, 6HzV _c _

2.67, dd, J = 2Hz, 8HzJ

1.93-1.63, m, C-5 methylene

1.63, s

1.40, s

1.23, d, H = 6.5Hz, C-IO methyl

Level F;

Preparation of 8-oxo-2 _J 2-diinethyl-7ö ^(c: - (lp-nitrobenzyl carboriyldioxyäthyl) -3-oxa-l-a2abicyclo- / 4.2, 7-t oc of

_ _{2 methyls}

N D

x:

70982 2/1020

new page 92

Under anhydrous conditions, a solution of δ-Οχο-2,2-dimethyl-7 «C- (l-hy droxyäthy I) -3-oxa-l-azabicy GlO-A, 2, o7-octane (60 mg, 0.302 mmol) in 0.6 ml of ether with treated with powdered potassium hydroxide (19 mg, 0.332 mmol), After 15 minutes, p-nitrobenzyl chloroformate (65 mg, 0.302 mmol) is added to the reaction mixture. One stirs another 15 hours at 25 ° C. The mixture is divided between pH 7 phosphate buffer and more ether. the The ether phase is washed with water and brine, dried over magnesium sulfate and filtered. When evaporating 67 mg of the filtrate under reduced pressure are obtained of a colorless oil. Purification by preparative thick layer chromatography over silica gel and developing with 1: 9 ethyl acetate / benzene gives 8-oxo-2,2-dimethyl-7 ^ C- (1-pnitrobenzylcarbonyldioxyethyl) -3-oxa-1-azabicyclo - / ^, 2,07-octane (40 mg) as a mixture of the diastereomers.

IR (CH ₂ Cl ₂ ) u: 5.68 (ß-lactam and carbonate), 6.19 and 6.54 (nitro)

KMR (CDCl ₃ ): 1.67, d, 2H, ArH

2.37, d, 2H, ArH

4.67, s, 2H, ArCH ₂

4.67-5.22, m, CH, CH

5.98-6.25, m, 2H, C-4 methylene

6.25-6.62, m, IH, C-6 methine

7.75-8.83, m, 2H, C-5 methylene

8.22, s, 3H, C-2 methyl

8.50-8.59, m, 5H, C-2 methyl + CH, CH

The 7 [beta] diastereoisomers or the 7 [deg.] C. and 7 [beta] mixtures are obtained in an analogous manner.

709822/1029

λλΑ

Level G:

Production of cis- and trans-3- (l-p-nitrobenzylcarbonyldioxyethyl) - * H (2-hydroxyethyl) -2-azetidinone

OR

tsf O

OH i

. "NH

8-Oxo-3-oxa-2,2-dimethy -1-jog (1-p-nitrobenzy lcarfconyldioxyäthyl) -l-azabicyclo / 3,2,07-oetan (I ₅ O g) v / ith 8 ml of acetic acid and 2 ml of water are dissolved and heated to 65 ° C. for 1.25 hours. The acetic acid and the water are removed under reduced pressure and the residue is taken up in benzene and evaporated, trans-3- (lp-Nitrobenzylcarbonyldioxyäthyl) - ^l i- (2-hydroxyethyl) -2-azetidinone as a mixture of the Obtains diastereoisomers.

KMR (CDCl ₃ )

5.67 Cβ-lactam), 5.72 shoulder, ", 2O and

6.57 (nitro)

1.73, d, 2H, H = 8.5 Hz, ArH

2, l »3, d, 2H, J = 8.5 Hz ₃ ArH

3.63, broad s, IH, NH

^, 37-5.13, m, IH, CH ₃ CH

4.72, s, 2H, ArCH ₂

6.07 - 6.53, m, IH, CH methine

6.23, t, 2H, J = 5.5 Hz, CH ₂ OH

6.73-6.93, m, .IH, C-3 methine

From 7.63 to 8.97, m, 3H, CH ₂ CH ₂ OH

8.53, d, J = 6.5 Hz, CH ₃ CH

709822/1029

AKi,

The cis-diastereoisomers or the cis-trans mixture is obtained in an analogous manner.

Steps D ', E', F ¹ and G ¹ as an alternative to steps D ₅ E ₃ P and G for the production of 3- (lp-Nitrobenzylcar bonyIdioxyäthyl) - * t- (2-hydroxyethyl) -azetidinone

OR

ti

R = -COCH

Levels D ', E ^j , F' and G *

Preparation of l- (2-tetrahydropyranyl) ^-1 i- / 2-tetrahydropyranyl ) -oxya'thyl7-2-azetidinone

Under nitrogen and at 25 C a solution of ⁱ i- (2-hydroxyethyl) -2-azetidinone (62 mg, 0.539 mmol) in 0.5 ml of anhydrous p-dioxane with 2 ₃ 3-dihydropyran

- 95 -709822 / 102Ö

(0.98 ml, 1.08 mmol) and p-toluenesulfonic acid monohydrate (19 mg, 0.10 mmol). The resulting solution is stirred for 60 minutes and then divided between 10 ml of a 0.5m pH 7 phosphate buffer and 10 ml of ethyl acetate. The aqueous phase is extracted a second time with ethyl acetate. The combined ethyl acetate solutions are washed with salt water, dried over magnesium sulfate and filtered. The filtrate is evaporated under reduced pressure, giving 216 mg of the crude product. Purification by preparative thick layer chromatography and developing with ethyl acetate gives 80 mg of 1- (2-tetrahydropyranyl) -il- / 2- (2-tetrahydropyranyl) -oxyethyl / 2-azetidinone as an oil. KIiR (CDCl ₃ ) y. 5.13-5.60, m, OCH

5.83-6.85, m, C-iJH + OCH ₂

6.95, dd, J = 5Hz and 15 Hz

7.35, dd, J = 3Hz and 15 Hz

7.62-8.95, m, CHCH ₂ CH ₂ CH ₂ CH ₂ +

C-3 methylene

Production of cis and trans-l- (2-tetrahydropyranyl) -3- (l-hydroxyethyl) -W2- (2-tetrahydropyranyl) -oxyethyl7-2-

azetidinone

If one works in the for the preparation of 8-oxo-2,2-dimethyl-7o £ - and ß- (1-hydroxyethyl) -3-oxa-1-azabicyclo- / i, 2.07-octane from 8-oxo-2,2-dimethyl-3-oxa-1-azabicyclo- / 3.2.0 / -octane described manner and uses l- (2-tetra-

- 96 -

709822/102 9

hydropyranyl) -4- / 2- (2-tetrahydropyranyl) -oxyethyl7-2-acetidinone, a diastereomeric mixture of both CIs- and trans-1- (2-tetrahydropyranyl) -3- (l-hydroxyäthy ¹ f- / 2 ~ (2-tetrahydropyranyl) -oxyethyl7-2-azetidinone.

Preparation of CIs- and trans-1- (2-tetrahydropyranyl) -3- (1-p-nitrobenzylcarbonyldioxyethyI) -4- / 2- (2-tetrahydropyranyl ) -oxyethyl7-2-azetidinone

/ SS

COCH,

■ HO.

If you work like in the production of 8-0xo-2,2-

dir.ethy ICC 1- (1-p-nitrobenzylcarbony ldioxyäthy l) -8-oxa-l- azabicyclo ^v / i, 2 ₃ 7-octane from 8-oxo-2 _J 2-dimethyl-7 - (1- hydroxyethyl) -3-oxa-l-azabicyclo- / i, 2, o7-octane and uses trans-1- (2-tetrahydropyrany1) -3- (1-hydroxyäthy1) -4- / 2- (2-tetrahydropyranyl) - oxyäthyl7-2-azetidinone, trans-1- (2-tetrahydropyranyl) -3- (lp-nitrobenzylcarbonyldioxyethyl) ^-1 } - / ^ - (2-tetrahydropyranyl) -oxyethyl7-2-azetidinone is obtained. The cis diastereoisomer is obtained in an analogous manner.

- 97 -

709822/1029

λλζ

Production of cis- and trans-3- (l-p-nitrobenzylcarbonyldioxyethyl) -4- (2-hydroxyethyl) -2-azetidinone

"R = COCH., - //.

A solution of trans-1 - ^ - tetrahydropyranyl) ^ - (1-pnitrobenzylcarbonyldioxyethyl) -4- / 2- (2-tetrahydropyranyl) -oxyethyl7-2-azetidinone in methanol at 25 ° C is with a 0.1 molar equivalent of Treated p-toluenesulfonic acid monohydrate. The solution is stirred for 2 hours and then neutralized with Im pH 7 phosphate buffer. The product is extracted into ethyl acetate. The ethyl acetate solution is washed with salt water, dried over magnesium sulfate and filtered. The filtrate is evaporated under reduced pressure to give trans-3- ^r (lp-nitrobenzylcarbonyldioxyäthyl) - 4- ⁱ receives azetidinone (2-hydroxyethyl) 2 ~. The cis diastereoisomer is obtained in an analogous manner.

- 98 - 7098 22/102 9

2657.676

AAf,

Level H:

Production of cis- and trans-3- (lp-nitrobenzylcarbonyldioxyethyl) -4- / 2 ₉ 2-bis- (2-hydroxyethyl) thioethyl? 2-

azetidinone

L-KH

/ ^ CHO

.NH

OH

I «

A mixture of anhydrous pyridine (0.1 * l6 ml, l, 8l mmol) and anhydrous, powdered chromium trioxide (92 mg, 0.916 mmol) in 8 ml anhydrous acetonitrile stirred for 30 minutes. 250 mg of dry material are added to the dark brown solution obtained Supercel given and then a solution of trans-3- (l-p-nitrobenzylcarbonyldioxyethyl) -4- (2-hydroxyethyl) -2-azetidinone (186 mg, 0.550 mmol) in 1 ml of anhydrous acetonitrile. After stirring for one hour, the Reaction mixture through a mixed packed bed 2 g each of silica gel and magnesium sulfate filtered.

- 99 -

709822/1029

The bed is washed repeatedly with a total of 30 ml of additional acetonitrile. The Piltrat is reduced under Pressure at 25 ° C concentrated to a volume of 3 ml. By thin layer chromatography (silica gel; Ethyl acetate / benzene 2: 1) it is found that this solution contains a product (R ~ = 0.38) that is less polar is than the starting material (R «= 0.21).

The acetonitrile solution of trans-3- (lp-nitrobenzylcarbonyldioxyethyl) ^-1 l- (2-oxoethyl) -2-azetidinone, which has been prepared as above, is mixed with 2-mercaptoethanol (0.386 ml, 5th , 5 mmol) and immediately thereafter treated with boron trifluoride etherate (0.176 ml, 1.3 mmol). After 15 minutes of stirring, this solution is divided between aqueous dipotassium hydrogen phosphate (1.5 g in H ml water) and 12 ml Ä'thylacetat. The aqueous phase is extracted a second time with ethyl acetate. The combined ethyl acetate solutions are washed with brine, dried over magnesium sulfate and filtered. The filtrate is evaporated under reduced pressure, 229 mg of an oil being obtained. The product is purified by preparative thick-layer chromatography over silica gel and developed with ethyl acetate, adding II8 mg of trans-3- (l ~ p-nitrobenzylcarbyldioxyethyl) -i <- / 2,2-bis- (2-hydroxyethyl) -thioethyl7-2- azetidinone is obtained as a colorless oil.

IR (CH ₂ Cl ₂ ) U: 5.75 (5.79 schilters) β-lactam and carbonate

6.20, 6.55 nitro

KMR (acetone-dg) T-: 1.70, d. J = 8.5 Hz, 2H, ArH

2.28, d, J = 8.5 Hz, 2H, ArH 2.118-2.88, m, IH, NH 4.63, s, ArCH ₂ λ _3H

^, 63-5.12, m, CH ₃ CH I

- 100 709822/102 9

5.80, t, J = 7 Hz, CH ₂ CH ^ g

5.80-7, ^ 5, m, C-HH + C-3H + SCH ₂ CH ₂ OH ^

7.63-8.33, m, 2H, CH ₃ CH

8.53, d, J = 6.5 Hz 3 H, CH ₃ CH

The cis diastereoisomers are prepared in an analogous manner. Alternatively, the mixed diastereoisomers obtained when a mixture of the diastereoisomers is used as the starting material.

- 101 709822/1020

Λ «

Level I:

Production of trans-3- (lp-nitrobenzylcarbonyldioxy ethyl) - * W2,2-bis (2-azidoethyl) -thioethyl7-2-azetidinone

2CH ₃ SO ₂ Cl

OCOOPiJB

To a solution of 211 mg (molecular weight = ^ 7 ^; 0.4 ^ 5 mmol) of trans-3- (lp-nitrobenzylcarbonyldioxyethyl) ^-1 i- / 2,2-bis- (2-hydroxyethyl) -thioethyl7-2- azetidinone in 5 ml of tetrahydrofuran (THF) (distilled from lithium aluminum hydride) at 0 ⁰ C 103 mg of mesyl chloride (molecular weight = H ¹ I 0.90 ¹ I
in mol) in 1 ml of tetrahydrofuran and immediately
then 13 ^ μΐ triethylamine (molecular weight 101; β = Ο _Λ 729;
0.967 mmol). The reaction mixture is stirred for 1 hour under Np. The triethylamine hydrochloride is found under N _?
filtered and washed with a few ml of additional THP. The clear, colorless filtrate is concentrated under a stream of Np and then pumped in a high vacuum for 10 minutes. The dimesylate is immediately dissolved in 5 ml of DMSO (distilled from CaHp at 8 mm and stored over a MA Linde molecular sieve) in the presence of 3 ^ 7 mg of NaN (molecular weight = 65; 5.3 ¹ l mmol). After stirring overnight and under N ₂ , 10 ml of H ₃ O and 20 ml of ethyl acetate (ΕΛ) are added. The layers are separated and the aqueous layer is washed three times with 10 ml of EA, each

- 102 -

709822/1029

AlO

The organic layer is washed once more with 10 ml of HpO and 10 nil of saline. The combined ethyl acetate layers are dried over anhydrous magnesium sulfate and filtered and concentrated under a stream of Np to give the crude diazide. Preparative thin-layer chromatography over silica gel gives trans-3- (1-pm.trobenzylcarbonyldioxyethyl) -W2 _J 2-bis (2-azidoethyl) -thioethyl7-2-azetidinone. The cis-diastereoisomers or the cis-trans mixture are obtained in an analogous manner.

Level J:

CO ₂ H

C (OH) ₂ ■ + 2N0 ₂ - <V

CO ₂ H

OCOOPNB

ToI. \

CO ₀ PNB

I ^l

C (OH) ₂

CO ₂ PNB

SCH ₉ CH ₉ N-

V '-ν

(CO ₂ PNB) ₂ j

PNB =

- 103 • 709822/1029

A freshly prepared (H. Davies and M. Schwarz, JOC, 30, 12! {2 (1965)) solution of p-nitrophenyldiazomethane (29 mmol) in 150 ml of ether is stirred into a solution of 1.0 g of oxomalonic acid monohydrate ( Molar weight = 136; 7.35 mmol) in 50 ml of ethyl acetate (EA) at ⁰ ° C. After 2 1/2 hours, the yellow solution is concentrated to about half its volume on a rotary evaporator with gentle heating, dried over anhydrous magnesium sulfate, filtered and concentrated to an oil as described above. To the crude p-nitrobenzyl ester in 50 ml of toluene (ToI.) Are added 3.5 mg of Tran3-3- (lp- * Nitrobenzylcarbonyldioxyäthyl) - ⁱ l- / 2,2-bis- (2-azidoethyl) -thioethyl7- 2-azetidinone (molecular weight = 52 ¹ l; 6.75 mmol). The reaction mixture is heated with stirring on an oil bath, about 1/3 of the toluene being allowed to distill off. Toluene (dried over a 3A l / 16 "Linde molecular sieve) is again added to make up the volume to 50 ml and the azeo-drying proi'-jSG is repeated three times. The solution is then refluxed under N ^ for one hour and the? The azeo-drying process is repeated one last time and the reflux treatment is continued for an additional hour. Concentrating the resulting solution under a stream of nitrogen gives crude 1. The crude material is chromatographed on silica gel to give 1-the cis-diastereoisomers or the cis-trans mixture is obtained in an analogous manner.

709822/1029

Level K:

κι%

OCOOPNB

SCH ₇ CH ₂ N ₃

.K OH

-f SOCl ₉ Pv

1 .

OCOOPNB

SCH ₀ CH ₀ N ₀

ZZO

SCH ₉ CH ₂ N ₃

Cl

4-

ag. DMF

OCOOPNB

SCH ₀ CH ₀ N
ZZ

(CO ₂ PNB),

- 105 -

709822/1029

^2552676

To a solution of 2.80 g of 1 (el Molgewi * = 912; 3.07 mmol) in 35 ml THF (distilled from Lithiutnaluminiumhydrid) at -20 ⁰ C 0.3 ml pyridine (molecular weight = 79; p = 0.932; 3.73 mmol) (distilled from NaH and stored over a ¹ JA Linde molecular sieve). While stirring under N ₂ , 0.1138 g of thionyl chloride (molecular weight = 119; 3.68 mmol) in 1 ml of TH? admitted. The reaction mixture is under N _? Stirred 5 minutes at -20 C and then 1/2 hour at 0 C and finally 1 hour at 25 C. The pyridine hydrochloride is under N _? filtered off and washed twice with benzene (dried over a 3A l / lo "Linde molecular sieve). The combined filtrates and washing solutions are concentrated under an Np stream, slurried in a small volume of benzene with anhydrous MgSO ₁₁ , filtered under Np and then concentrated under an Np stream ". An oil is obtained by pumping under a high vacuum for 1/2 hour. To this freshly prepared chlorine compound, O _3, 885 g of triphenylpho-ophine (Kolec weight = 262; 3 * 38 mmol) in 66 ml of 9: 1 dimethylformamide (DK?) / HpO and then 550 mg are added with stirring

K ₂ HPO ₁ , (molecular weight = Uk; 3.16 mmol). The reaction mixture is stirred at 25 ° C. for 35 minutes. After diluting with SA and saline, the layers are separated and the aqueous layer is extracted three times with EA. The combined EA layers are washed with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under a stream of Np to give crude 2. The material is chroma-

r * J

tographiert, where 2 is obtained, The cis-diastereoisomers or the cis-trans mixture is obtained in an analogous manner.

- 106
709 8 22/1029

Level L:

OCOOPNB

/ TM

+ Br

. «Η

OCOOPNB

.V.

O'

Br Br

(D

(2) KaH, DMF

OCOOPNB f

<y

SCH ₂ CH ₇ Ii ₃

(CO ₂ PNB) ₂ SCH.

Br

- 107 -

709822/1029

To 7.8 ml of pentane (dried over a 4 A Linde molecular sieve) 0.2 ml of Br (molecular weight = ΙβΟ; Z- 3.12; 3.9 mmol) are added. To a solution of 950 mg ^ (molecular weight = 896; 1.05 mmol) in 15 ml Diäthylather (dried over 3A l / lo "Linde molecular sieves) 2.3 are added at 0 ⁰ C and added dropwise with stirring ml Np of the above 0.49m Br ₃ solution (1.13 mmol). After stirring for 10 minutes at 0 ° C., 114 μl of cyclohexene (molar weight = 82, S- 0.81; 1.13 mmol) are added Add 53 kg of 57% NaH (57 % of 53 mg = 30.2 mg, molecular weight 24, 1.26 mmol) in mineral oil to the stirred reaction mixture anhydrous CaSOj, at 40 mm and stored over a 4A Linde molecular sieve). Stirring is ^{continued for 3 hours under Np at 0 °} C. The reaction mixture is poured into a stirred ice-cold mixture of 2.5 ml Im. KH 40 ml IL, 0 -75 ml EA .. After separating the layers, the aqueous layer is saturated with NaCl and extracted again with EA. The combined organic layers are washed once with saline solution ext Rahiert, dried over anhydrous MgSOh, filtered and concentrated under a stream of Np and then pumped in a boiling vacuum j, Vi whether the crude 3 is obtained. Preparative thin post chromatography over silica gel gives 3. The cis-diastereoisomers or the cis-trans mixture are obtained in an analogous manner.

709822/1029 ^lo8 "

Level M: Al (o

OCOOPNB

SCH ₉ CH ₉ N _n

¹ p

(GO ₉ PNE -f Br ₂ ether

OCOOPNB

BSiWr

Br CHCH '

(CO ₂ PNB) ₂ + NaH

ΏΜΡ

0C00PN3

SCH ₂ CH ₂ N ₃

- 1Oq -

709822/102 9 '

To 9s 16 nil pentane (dried over an HA Linda molecular sieve) is added 0.2 ml of Br ₂ (molecular weight = 160; 3.9 mmol), To W mg 3_ (molecular weight = 793; 0.598 mmol) in 13 ml of diethyl ether (dried over 3A 1/16 "Linde molecular sieves) at 0 C under Np is added dropwise with stirring 1.52 ml of the above 0, ⁱ t2m _Br? solution (0.63 mmol). ■ After 15 minutes at 0 ° C 33 mg 57 £ NaH (57% of 33 mg = l8,8 mg; molecular weight = 2h; 0.78 mmol) was added and immediately thereafter are added 6.35 ml of ice-cold DMF (distilled from anhydrous CaSO ₂ at ¹ IO mm. and stored over an HA Linde molecular sieve) The reaction mixture is stirred for 1 1/2 hours at 0 ° C., then _{poured into a stirred, ice-cold mixture of 1.6 ml Im KH PO ^ -20 ml H 2} O and 20 ml EA. The layers are separated and the aqueous layer is saturated with NaCl and extracted again with more EA. The combined organic layers are washed once with brine, dried over ana pure magnesium sulfate and filtered. The filtrate is concentrated under a stream of Np and then placed under high vacuum to give the crude H. Preparative thin-layer chromatography over silica gel gives 4. The cis-diastereoisomers or the cis-trans mixture is obtained in an analogous manner.

- 110 _
7098 22/1029

Stages:

OCOOPNB

0C00PN3 '. !

. I.

In

(CO ₂ PNB) ₂

To 210 mg Ji (molecular weight = 87I; 0.241 mmol), dissolved in O _3, 5 ml DMSO (distilled from CaH ₂ at 8 mm and stored over a hA Linde molecular sieve) add HO mg 1,5-diazobicyclo with 25 C * urfer stirring - / 5.4, o7-undec-5-en (distilled at 80 ° C / 2 mm) (molecular weight = 152; 0.263 mmol) in 0.7 ml of dimethyl sulfoxide (DTiSO). The solution is stirred for h hours under Np and then added to a stirred, ice-cold mixture of 0.48 ml Im KHpPO ^, 7 ml HpO and 10 ml EA. After separating the layers, the aqueous layer is extracted again with EA. The combined organic layers are washed once with salt water and then dried over anhydrous magnesium sulfate, filtered, and under a stream of Np

- 111 -

709822/1029

concentrated and then placed under a high vacuum, the crude 5 being obtained. By preparative thin layer chromatography over silica gel one obtains 5 · The cis-dia-

f \ J

stereoisoir.eren or the cis-trans mixture is analogous Way received.

Level 0:

s-collidine

(CO ₂ PNB) ₂

To a solution of 187 mg 5 (molecular weight = 791;

rs »

O ₃ 236 mmol) in 2.5 ml of s-collidine (distilled from powdered KOH at 30 mm pressure) are H5 mg of anhydrous LiI (dried for a few hours at 100 ^° C. over P _? 0 under vacuum). (Molecular weight = 13 ^ ; 0.330 mmol) given. While stirring under N _? the reaction mixture is heated to 120 ° C. on an oil bath. After a total of 25 minutes, the reaction mixture is cooled to 25 ° C., diluted with CHpClp and transferred to a round bottom flask in order to concentrate under a stream of Np and in a high vacuum. The residue

- 112 -

709822/1029

Λ "JO

is divided between 10 ml EA and 1.8 ml Im KH ₂ PO ₄ in 10 ml H ₃ O and then the aqueous layer is extracted two more times with EA. The combined organic layers were extracted with brine, dried over anhydrous magnesium sulfate, filtered and concentrated under a stream of nitrogen to give crude 6. Preparative thin-layer chromatography over silica gel gives 6, the cis-diastereoisomers or the cis-trans mixture is obtained in an analogous manner.

Level P:

OCOPNB

C0 ₂ ? N3

DMSO

SCH ₂ CH ₂ N ₃

To a solution of the mixed diastereoisomers 6 molecular weight = 612; 0.055 mmol) in 0.2 ml DMSO (distilled from CaHp at 8 mm and stored over an HA Linde molecular sieve) are added with stirring 9.5 pi 1,5-di-

S-en (distilled at -80 ° C / 2mm) (molecular weight = 152; £ = 1; 0.0525 mmol). The solution

709822/1029

- 113 -

26Β2676

is stirred under nitrogen for 15 minutes and diluted to a total volume of 1 ml with CHCl-, and immediately placed on 2 to 1000 u silica gel plates. The gang of

Product shows 7 as a mixture of cis and transit

diastereoisoiaeren.

Level Q:

OCOoPNB

OH

.CQ ₂ PNB

H ₂ / 2.8 kg / cn ²

In the presence of 61 mg PtOp, Gl mg 7 (molecular weight 612; 0.1 IrMoI) in 6 ml dioxane, 6 ml THF, 3 ml H ₀ O are hydrogenated with hydrogen for 4 hours at a pressure of 2.8 kg / cm. The reaction mixture is filtered through a Celite filter and _{washed with 2 ml of O 9} In pH J phosphate buffer. After concentrating in vacuo to the cloud point, the aqueous mixture is extracted with ethyl acetate. The aqueous layer is concentrated to a small volume and applied to a column of 100 g of XAD-2 resin. When eluting with HpO and discarding the initial fractions, the fractions which contain the product are freeze-dried, with 8 being obtained as a mixture of cis and trans diastereoisomers.

709822/1 Oi

The procedure described below for the enzymatic N-deacylation of thienamycin is applicable for all isomers of thienamycin, especially for the pronounced N-acetyl isomers 89OA and 89OA- ,, die are described below.

Peacetylation of N-acetylthienamycin

A l ^ ige (weight / volume) suspension of fertile Turf soil is prepared by adding 1 g of the turf soil to 100 ml of sterile phosphate buffer saline, v / obei the phosphate buffer saline solution has the following composition: ".

Phosphate buffer saline solution
NaCl 8.8 g

In phosphate buffer, pH 7.5 ^ 10 ml

distilled water 1000 ml

* In phosphate buffer, pH 7.5

16 ml Im KH ₂ PO ₁ J are mixed with SH ml Im K ₂ HPO ^. the pH of the phosphate buffer is adjusted by adding a small amount of either Im KHpPO ,. or Im KpHPOj, set to 7.5.

Aliquots of this 1 / yl storage soil suspension are made up to prepare 10x, 100x and 100x dilutions.

1 ml portions of the stock suspension or 1 ml portions of 1ox, Loox and lOOOx dilutions are added to 2 ml portions of sterile!, Above Agarlösungen at ^ 8 ⁰ C. The mixture is quickly poured over the surface of sterile Petri dishes 85 mm in diameter containing 20 ml of Medium A. Medium A has the following composition:

- 115 -

709822/1029

Medium A 3.0 ε KH ₂ PO ₄ 7.0 ε K ₂ HPO ₄ 0.1 ε MgSO ₄ 1000 ml distilled H ₃ O 8.5 ml N-acetylethanol
amine solution N-acetylethanol solution

N-acetylethanolamine is lox diluted in HpO and membrane sterilized. This solution is added after the autoclave treatment. For solid media: 20 g of agar are added.

The Petri dishes are incubated at 28 ° C. for 18 days. A well isolated colony is picked and spread on a Petri dish containing Medium B. Medium B has the following composition:
Medium Β

te ^ O g ^!

ground wheat flour 15 g
desfc. V / ater 1000 ml

pH: adjusted to pH 6 using NaOH. For solid media: add 20 g of agar.

A single culture is selected and kept at 28 C for 2 days grown on an agar culture of medium B. Part of the growth of this culture is on the surface of 6 cultures made from medium 3 are spread out. These cultures are incubated at 28 ° C. for 2 days. This culture has been identified as Protaminobacter ruber and designated MB-3528 in the culture collection from Merck & Co., Ine, Rahway, N.J., USA and a sample has been deposited with the Agricultural Research Service, U.S. Department of Agriculture, Accession No. NRRL B-143.

- 116 -

709822/1029

Part of the growth of the agar culture of Protaminobacter rub er MB-352 8 is used to remove a 250 ml Erlenmayer flask, containing 50 ml of medium C, to inoculate. Medium C has the following composition:

Medium C

Dextrose 20 g

Pharmamedia 8 g

Soaked corn liquid (wet base) 5 g

distilled water 1000 ml

pH: adjusted to 7 with NaOH or HCl N-acetylethanolamine solution 8.5 ml

N-acebylethane olamine solution

N-acetylethanolamine is diluted 1Ox in HpO

and membrane sterilisable. This solution is found after the Provide autoclave treatment.

The flask is shaken on a shaker at 28 ° C and 220 rpm for 4 days. A 25 μl portion from the flask is centrifuged at 8000 rpm for 15 minutes. The excess is removed and the cells on the surface of the solid medium are scraped off and added to a 0.5 nil 0.05 M potassium phosphate buffer, pH 7, ^. The suspension obtained is subjected to ultrasound treatment using a Branson Instruments Model LS-75 Sonifier with a 1/2 inch probe at a setting J- ¹ with four 15-second Intervallena while the suspension during and cooled 'between interruptions in Eisvrasser . A 10 μl Anta.l of the sonicated product is mixed with 25 μl of solution containing 8 * 10 pg / ml N-acetylthienamycin in 10 mmol potassium phosphate buffer, pH 7, and incubated at 28 ° C. overnight. Controls, x-relche contain the antibiotic and buffer alone, and sonicated cells and buffer

- 117 709822/1029

no antibiotic were also made. After an incubation 2 μl amounts are increased overnight at 28 ° C cellulose-coated thin-layer chromatography plates that were developed in ethanol: HpO, 70:30. After air drying, the thin-layer chromatography plate is used placed on a plate infected with Staphylococcus aureus ATCC 6538P for 5 minutes.

The infected plates were prepared as follows: The over-growth of the organism used for the infection Staphylococcus aureus ATCC 6538P in meat broth plus 0.2 % yeast extract is diluted with meat broth plus 0.2 % yeast extract to a suspension which has a 60% Has good transmittance at a wavelength of 660 nm. This suspension is added to Difco nutrient agar, supplemented with 2.0 g / l Difco yeast extract, at 37 to 8 ° C. in order to obtain a composition which contains 33 _s 2 ml of the suspension per liter of agar. HO ml of this suspension are placed on Petri dishes, 22.5 cni χ 22.5 cm. poured and these plates are cooled and held at 4 C until used (5 day maximum).

The thin layer chromatography plate is removed and the. The plate containing the organism is incubated at 35 ° C. overnight. In addition to the unreacted bio-active N-acetyl-thienamycin'mit a point at R "from 0.7 to 0.89, a bio-active point observed in R ~ O ₄ ¹ I ¹ IO, Hj, which is attributed to the thienamycin . Control incubation mixtures of the antibiotic plus buffer, sonicated cells plus buffer, and antibiotic plus buffer to which sonicated cells had been added shortly before thin layer chromatography application showed no bioactive material at R _f 0.1 - 0.47.

- 118 709822/102 9

Production of 89OA ₁ , a special isomer

OH

.N.

COOH

A tube of a freeze-dried culture of Streptomyces flavogriseus MA-4i} 34 (NRRL 8139) is added opened aseptically and the contents are placed in a tube suspended, which contains 0.8 ml of sterile Davis salt of the following composition:

Davis salt

Sodium nitrate 0.5 ε K ₂ HPO ₁ J. 7.0 ε KH ₂ PO ₁ J 3.0 G (WH ₁ Y) _{2 SOk} 1.0 G MgSO _r 7H ₂ O 0.1 G distilled water . 1000 ml

This suspension is used to incubate four cultures of medium A with the following composition:

Medium A

Glycerin 20.0 g

primary yeast 5.0 g

Fish meal 15.0 g

distilled water 1000 ml

Agar 20.0 g

pH: adjusted to 7.2 using NaOH

The inoculated cultures are a V / o before incubated at 27-28 ° C and then stored at 4-6 ⁰ C until use.

- 119 709822/1029

A 1/3 serving of the growth of three cultures is used to inoculate 9 stoppered Erlenmeyer flasks containing 50 ml of Medium B of the following composition:
Medium B

Yeast on colysate (Ardamire) 10.0 g

Glucose x 10.0 g

Phosphate buffer ⁺⁺ 2.0 ml

₁₁ -TH ₂ O 50 mg

distilled water 1000 ml

pH: adjusted to 6.5 using HCl or NaOH

Ardamine: Yeast Products Corporation

Phosphate buffer solution

KH ₂ PO ₄ 91.0 g

Na ₂ HPO ₄ 95, above

distilled water 1000 ml

The flasks containing the germs are shaken for 1 day at 27-28 ° C. on a shaker at 220 rpm. The flasks and the contents are stored at 4 ^° C. for 1 day.

2-1 Erlenmeyer flasks, each containing 250 ml of Medium C, are inoculated with 8 ml per flask of growth from the flask containing the germs. Medium C has the following composition: Medium. C.

Tomato paste 20.0 g

primary yeast 10.0 g

Dextrin (Amideχ) 20.0 g

CoCl ₂ .6H ₂ O 5.0 mg

distilled water 1000 ml

pH: adjusted to 7.2-7 _a b by MaOH

- 120 709822/1029

After the inoculation, the production flasks are incubated at 2 h C with shaking on a shaker operated at 212 rpm for h days. The flasks are then emptied and the contents assayed for activity using standard Salmonella gallinarum 1431287 and Vibrio percolans ATCC 84ol plates using 1.25 cm disks which were immersed in the centrifuged broth samples. The samples are diluted with 0.02m phosphate buffer, pH 7.0, if necessary. The results are given below:

Time of harvest (Harvest Age

hours) h 96

pH 7.2

Salmonella gallinarum (mm zone) 29.5

Vibrio percolans 1/10 dilution

(mm zone) 31

89O test units 40

7 liters of the entire fermentation broth are cooled to 3 ° C and in 200 ml portions at 900 rpm for 15 minutes each centrifuged.

To the combined supernatant liquids _{70 am} neutral EDTA are added and the whole sample is placed on a Dowex-1 χ 2 (Cl), 50-100 mesh column with bed dimensions of 5> 1 × 25 cm and a flow rate of k0 ml / min. The column is comprising ml, washed with 500 ml of deionized water, 5 in Tris-HCl buffer, pH 7 ₃ 0, and 25 juMol neutral EDTA. The antibiotic is eluted with 1 liter of deionized water containing 50 g of sodium chloride and the column is then washed with 300 ml of deionized water. Fractions of 100 ml v / earth collected, starting from the first appearance of the salt at the column exit. The bio-activity lies in the fractions "

- 3 21 -

709822/1029

up to 10 with a peak at fraction 2. Fractions 2 to 5j which contain 17% of the applied activity are pooled.

The pooled fractions are concentrated to 110 ml using a rotary evaporator under reduced pressure and the pH is adjusted to 5.8 by adding 4.2TnI Im HCl. The concentrate adjusted in this way is applied to a column of XAD-2 with a bed dimension of 3.8 χ 50 cm, which had previously been washed with 3 liters of 60% lgam aqueous acetone (volume / volume) and then with 3 liters of deionized water, 3 1 of 5 * (weight / volume) sodium chloride and 1 1 25 * (weight / volume) sodium chloride in deionized water. The concentrate used is allowed to flow off to the level of the bed. The antibiotic is eluted with deionized water at a flow rate of 15 ml / min. 22 fractions of 100 ml each are collected from the first application of the sample. Bio-activity appears in fractions 6 to 22 with a peak at fractions 9 and 10. Fractions 9 to 20 are pooled for further processing. Fractions prepared similarly are combined and the combined fractions are concentrated to 70 ml using a rotary evaporator under reduced pressure.

The concentrate is applied to a Dowex-1 χ H (Cl ") työO mesh column with a bed dimension of 2.2 χ 27 cm. The column is washed with 50 ml of deionized water and the antibiotic is eluted with 2 1 0 _a 070m NaCl + 0.005m NH ₁ (Ci + 0.0001m HH-, in deionized water at a flow rate of 2 ml / min. Fractions of 9.5 ml are collected.

- 122 7 0 9 8 2 2/1029

The main peak of the antibiotic is eluted into the Fractions 142 to I63. Fractions 146 to 157 are combined for further processing.

The combined fractions 146-157 are concentrated to 3 ml by evaporation under reduced pressure on a Rotary evaporator and. the concentrate is made by adding 5 ul Im MH-. brought to pH 6.5. The concentrate will be on a column (2.2 χ 70 cm) of a Bio-Gel P-2 (200 to 400 Mesh), which had been washed beforehand with 20 ml of 5m NaCl and 500 ml of deionized water. After this the concentrate had run off to bed level 2 rinses made with ImI deionized water each time and again drained to bed level. The column is then eluted with deionized water a flow rate of 0.6 ml / min. Fractions of 2.9 ml are collected.

The main peak of the antibiotic is eluted in fractions 63 to 70. Fractions 64 and 65 are used for the rest Give ζ us the amount of work-up.

The combined fractions 64 and 65 are on a thin film evaporator concentrated under reduced pressure to 2 ml and then frozen in an envelope and lyophilized for 8 hours in a 14 ml screw cap provided vessel, with 2.25 mg of the essentially pure antibiotic 89OA. receives. The N-Acety! Group v ^ ird cleaved off as previously described, giving the free base:

- 123 709322/1029

AT THE

Manufacture of 89OA ,, a special isomer

OH

t

Il

J N LLjjOOH

-S LLjj

One tube of a lyophilized culture of Streptomyces flavogriseus MA-4434 '(MRRL 8l., 3) is opened aseptically and the contents are suspended in a tube containing 0.8 ml of a sterile Davis saline solution following composition:

Davis-SaIz

Sodium citrate 0.5 g

K ₂ HPO ₁₁ 7.0 g

KH ₂ PO ,. 3.0 g

(NH ₁ J) ₂ SO ₁ , 1.0 g

MgSQ ₄ .7H ₂ O 0.1 g

distilled water 1000 ml

This suspension is used to make 4 cultures of the medium. A. to inoculate the following composition:

Medium A

Glycerin 20.0 g

primary yeast 5.0g

Fish meal 15.0 g

distilled water 1000 ml

Agar 20.0 g

pH: adjusted to 7.2 under NaOH

709822/102 9

The inoculated cultures are incubated for 1 week at 27-28 ° C and then stored at 4-6 ° C until used (no longer than 21 days).

A 1/3 proportion of the growth from 4 cultures is used for inoculation of 12 stoppered 250 ml Erlenmeyer flasks containing 50 ml of medium B of the following composition was used :

Medium B

Yeast autolysate (Ardamine) 10.0 g

Glucose 10.0 g

Phosphate buffer ⁺⁺ 2.0 ml

MgSO ₄ .7H ₂ 0 5 0 mg

distilled H ₃ O 1000 ml

pH: adjusted to 6.5 under HCl or NaOH

Ardamine: Yeast Products Corporation

Phosphate buffer solution ■ KH ₂ PO ₄ 91.0

₂ PO ₄

he

Na ₂ HPO ₄ 95.0 g

distilled water 1000 nl

The bottle with the germs is shaken for 1 day at 27-28 ° C. on a vibrating machine at 220 rpm. The bottle and ' the contents are kept stationary for 1 day at 4 ° C.

44 2-1 Erlenrceyer flasks, each containing 200 ml of Medium C, add 8 ml per flask of growth from the inoculated flasks. Medium C has the following Composition:

Medium C

Tomato paste 20.0 g

primary yeast 10.0 g

Dextrin (Amidex) 20.0 g

0 9 8 2 2/1029

26526/6

CoC1 ₂ .'6H ₂ Ö 5.0 mg

distilled water 1000 ml

pH: adjusted to 1.2 I ₃ H with NaOH

After inoculation, the production flasks at 2H H days and 5 hours machine on a chute! Operated with 212U / min, shaken. The flasks are then emptied and checked for activity using standard Salmonella gallinarum MB1287 and Vibropercolans ATCC 8461 sample plates using 1.25 cm sample disks which were immersed in the centrifuged samples of the broth. The samples are diluted with 0.2 M phosphate buffer, pH 7.0, if necessary. The results are given below:

Time of harvest (Harvest

Age hours) h 101

pH 7.2

Salmonella gallinarum (mm zone) 35 Vibrio percolans 1/10 dilution

(now zone) - J> H

890 trial units 121

This fermentation becomes a total of 7 l of the total Broth obtained and these are cooled to 3 C and centrifuged in 200 ml portions at 9000 rpm during each 15 minutes. 1.7 ml of 0.1 M neutral EDTA are added to the combined supernatant solutions and the The batch is kept at 3 ° C.

The above fermentation is repeated under identical conditions with the exception that the HH 2-1 Erlen'meyer flasks are inoculated with 7 ml per bottle of growth from the bottles containing the cultures; the pH and the test results obtained are given below:

- 126 709822/1023

101 3 26526/6 Time of harvest (Harvest
Age hours) h 7, pH 38 Salmonella gallinarum (rr.n zone) 39 8th Vibrio percolans 1/10 dilution
(mm zone) 92, 890 trial units

A total of 7, ^ 1 of the total broth resulting from this fermentation have been obtained, are cooled to 3 C and each 15 minutes in 200 ml portions at 900 U / Hin centrifuged. 1.8 ml of 0.1m neutral EDTA are added to the combined supernatant solutions.

The supernatant from the centrifuge broths like it obtained from the two fermentations mentioned above in this example is combined, where nan is a total volume of 13 receives 1.

The combined supernatant solutions are passed through a column of Dowex-1 χ 2 (Cl) ₃ 50-100 meshes with a bed dimension of 4 ₃ 7 cm χ 50 cm and a flow rate of 60 ml / min. The column is washed with 1 l of deionized water and the antibiotic is eluted with 5 l of a 5% (weight / volume) NaCl solution containing 0.01 M Tris-HCl buffer, pH 7.0, and 25 mol EDTA. Fractions of 220 ml are collected at a flow rate of 50 ml / min and the fractions are checked against Salmonella gallinarum MB 1287 plates.

Antibiotic activity is present in fractions 3 to 26 with a peak in fractions 5 and 6. Fractions 5 to 9 are combined for further processing. The * pH of the pooled fractions is 10.8 and the combined fractions contain 2 ¹ I% of the initial bioactivity, as measured against Salmonella gallinarum MB1287 ~ plates.

709822/1029

The combined fractions 5 to 9 are concentrated to 150 ml by means of a rotary evaporator under reduced pressure and applied to a column ( ¹ l _s .9 cm χ * f7 cm) of XAD-2, which had previously been washed with 5 1 6O / 5igem (Volume / volume) aqueous acetone and then with 5 1 deionized water and 5 1 50 g / l NaCl in deionized water. The sample is applied in 20 ml portions, soaking the column to the bed level each time. When use is complete, three 20 ml portions of deionized water are added and allowed to flow to bed level each time. The sample is eluted with deionized water at a flow rate of 20 ml / min. All measures in which the XAD pillar is used will be

ο carried out at room temperature (2 ¹ JC) and the eluting fractions are quickly cooled in an ice bath immediately after collection. Fractions of 95 to 230 ml are collected.

The antibiotic activity appears in fractions 2 to 2I ₃ as measured from a sample of Salmonella gallinarum KB1287 plates, with a peak at fractions 5 to 7 (which account for 510 to 895 ml of eluted volume, calculated from the first application-deionized Water). Fractions β to 21 are combined.

The combined fractions 6 to 21 are concentrated to 68 ml under reduced pressure on a rotary evaporator and then diluted to 112 ml by adding deionized water. The concentrate is applied to a column (2.2 cm × 21 cm) of Dowex-1 × ¹ J (Cl ") ¹ 100 mesh. The column is washed with 20 ml. Deionized water and the antibiotic is eluted with 2 1 0 , 07m NaCl + O ₃ 005m MHi ₁ Cl + Ο, ΟΟΟΙίπ NH ₇ in deionized water at a flow rate of 1.6 ml / min. Fractions are

709822/102 9 " ¹²⁸ ~

collected from each 8 ml. Fractions 157 to 179 have the greatest activity and are given together.

These fractions given from the amines are concentrated to 7 ml on a rotary evaporator under reduced pressure, the pH is adjusted to 7.5 by adding 20 μl of 1M NaOH. The solution is further concentrated to 5 ml and applied to a column (2.2 × 75 cm) of Bio-Gel P-2, 200 to 400 Maschan. The sample is washed in the column bed with two washes of 1 ml of deionized water each and eluted with deionized water at a pleating rate of 0.6 ml / min. Ten fractions of 3 ₉ 3 ml and then 60 fractions of 2.65 ml and ten fractions of 2.0 ml are collected.

The fractions are adjusted to the pH range between 7 _a 5 and 8.0 by adding 1.5 to 2.5 μΐ 0.1m NaOH. The fractions 62, 63, 6'4 and 65 are frozen and lyophilized individually in 14 ml glass tubes and stored at -20 ° C under vacuum.

- 12Q -

70 9822/1029

To isolate the antibiotic 89OA-, from 89OA ₁ , one makes use of the advantage of the relatively higher resistance of the antibiotic 89OA., To degradation by penicillinase in the following way:

Bio-Gel fraction 63 is combined with fractions 61, 66 and 67 from the Bio-Gel column. United to these four Fractions are 0.2 ml in Tris-HCl buffer, pH 7.5, and 0.2 ml of penicillinase (Difco "Bacto-Penase"). After 113 minutes at 23 ° C., a further 0.2 ml of penicillinase is added. After a further 7 hours at room temperature another 0.2 ml of penicillinase are added, and after a further 2 hours at room temperature, the The reaction mixture was cooled on an ice bath. The finished one The reaction mixture is made up to 15 ml by adding 5 ml deionized Viasser.

The reaction mixture is adsorbed on a Doweχ χ Ir (Cl "*) ¹ IOO mesh column, bed dimension 2.15 cm χ ¹ JO. The antibiotic 89OA is eluted with 0.07M NaCl + 0.005M NIKCl + 0.0001M NH, in Deionized water at a flow rate of 3 ml / min. Fractions of 13.8 ml are collected in each case. Fractions 187 to 200 times are put together.

- 130 7 0 9 8 2 2/1029

The combined fractions are concentrated to ¹ l ml in a rotary evaporator under reduced pressure and the concentrate is applied to a column (2.2 × 70 cm) Bio-Gel P-2, 200-600 mesh. The antibiotic 89OA., Is eluted with deionized ater V7 with a Pliessgeschwirtdigkeit 0 ₃ 6 ml / min. 30 fractions of 3.3 ml and then 50 fractions of 2.65 ml are collected. .

Fractions 66 to 70 are united and the united Samples are concentrated to 1.5 ml under reduced pressure using a rotary evaporator and the concentrate is made frozen and freeze-dried, giving 5, ^ mg of one Solid containing the antibiotic 89OA-. and the rest Contains salt. The M-Acety! Group has split off, as previously described, so that you get the free base:

OH
\ ί
N if COOH Λ

The antibiotic thienamycin and its preparation are described in DT-OS 25 52 638. It is available from Cultivation of the microorganism Streptomyces cattleya (MA-429'7), one of which is in the culture collection of Northern Regional Research Laboratories, Northern Utilization Research and Development Division, Agricultural Research Service, U.S. Department of Agriculture, Peoria, Illinois, V.St.A. deposited and is designated NRRL 8057.

709822 / 1029-131-

Claims (22)

15833T "Patent Claims 1. Compounds of the following general formula: OH -N- ₂ CH ₂ COOH and their pharmacologically acceptable salts, ethers and esters, where R, R and R each independently of one another C fr rj Hydrogen atom (not all radicals R, R and R are simultaneously hydrogen atoms), a substituted or unsubstituted lower alkyl radical with 1 to 10 carbon atoms, alkenyl radical with 2 to 10 carbon atoms or alkynyl radical with 2 to 10 carbon atoms, a ring-substituted or unsubstituted cycloalkyl, cycloalkenyl, cycloalkenylalkyl or cycloalkylalkyl radical with 3 to 6 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain, an aryl radical with 6 to 10 carbon atoms, an aralkyl radical with 6 to 10 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain or a mono- or bicyclic heteroaryl or heteroaralkyl radical with 4 to 10 ring atoms, of which one or more (one) oxygen, nitrogen and / or Represent sulfur atom (s), and 1 to 6 carbon atoms - 132 709822/1029 15833Y in the alkyl chain, the ring or chain substituents each (at least) one chlorine, bromine, iodine or Fluorine atom, an azido, cyano or amino group, a mono-, di- or trialkyl-substituted amino group (where the alkyl radical has 1 to 6 carbon atoms), a hydroxyl group, an alkoxy radical with 1 to 6 carbon atoms, a thioalkyl radical having 1 to 6 carbon atoms, a carboxyl or oxo group, an alkoxycarbonyl radical with 1 to 6 carbon atoms in the alkoxy moiety, an acyloxy radical with 2 to 10 carbon atoms, a carbamoyl group, a mono- or dialkylcarbamoyl group (the alkyl radicals each having 1 to 4 carbon atoms), a thiocyanate group (-SCN) or a nitro group represent. 2. Compounds according to claim 1, characterized in that R ft V
R ⁵ , R and R 'each represent a hydrogen atom or a lower alkyl radical having 1 to 6 carbon atoms. 3. A compound according to claim 2 having the following formula: SCH ₂ CH ₂ NHCH ₃ COOH 4. A compound according to claim 2 having the following formula: SCH ₂ CH ₂ N (CH ₃ ) COOH - 133 - 70982 2/102 9 15833Y 5. Compounds according to claim 1 with the following general Formula: SCH ₂ CH ₂ N (CH ₃ J ₃ A Θ COOH in which A ^ is a pharmacologically acceptable anion. 6. Compounds of the following general formula: .4 SCH ₂ CH ₂ NR ⁵ R ⁶ R COXR and their pharmacologically acceptable salts, where R, R and R independently of one another each represent a hydrogen atom (not all radicals R- ⁵ , R and R ^ are hydrogen atoms at the same time), a substituted or unsubstituted lower alkyl radical with 1 to 6 carbon atoms, alkenyl radical with 2 to 10 carbon atoms or an alkynyl radical with 2 to 10 carbon atoms, a ring-substituted or unsubstituted cycloalkyl, cycloalkenyl, cycloalkenylalkyl or cycloalkylalkyl radical with 3 "to 6 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain, an aryl radical with 6 to 10 Carbon atoms, an aralkyl radical with 6 to 10 ring carbon atoms and 1 to 6 carbon atoms in the alkyl chain or a mono- or bicyclic heteroaryl or heteroaralkyl radical with 4 to 10 ring atoms, of which one or more (one) oxygen, nitrogen and / or sulfur atoms (e) represent, and 1 to 6 carbon atoms in the alkyl chain, the ring or Chain substituents each have (at least) one chlorine, bromine, iodine or fluorine atom, an azido, cyano or amino group, a mono-, di- 709822/10 2-9 _ _ or trialkyl-substituted amino group (where the alkyl radical Has 1 to 6 carbon atoms) »a hydroxyl group» an alkoxy radical with 1 to 6 carbon atoms »a thioalkyl radical with 1 to 6 carbon atoms, a Carboxyl or oxo group, an alkoxycarbonyl radical with 1 to 6 carbon atoms in the alkoxy moiety, an acyloxy radical with 2 to 10 carbon atoms »a carbamoyl group, a mono- or dialkylcarbamoyl group (where the Alkyl radicals each have 1 to 4 carbon atoms), represent an Ihiocyangruppe (-SCH) or an Nltrogruppe, Ir is a hydrogen atom or an acyl, alkyl, aryl, aralkyl, alkenyl or alkynyl radical, Σ represents an oxygen or sulfur atom or a radical NR ¹ (R ¹ = H or R) and R is a hydrogen atom »an alkyl radical with 1 to 10 carbon atoms» a phenacyl group or ring-substituted phenacyl group · where the substituent is a chlorine »bromine or Is fluorine atom or an alkyl radical having 1 to 6 carbon atoms, an alkoxyalkyl radical, the alkoxy moiety is open-chain or cyclic and has 1 to 6 carbon atoms and the alkyl portion also has 1 to 6 carbon atoms »an alkanoyloxyalkyl radical with 2 to 12 Carbon atoms, a halogen or perhaloalkyl radical, wherein the halogen atom (s) is (are) chlorine, bromine or fluorine atom (s) and the alkyl chain is 1 to 6 carbon has atoms, an alkenyl radical with 2 to 10 carbon atoms, an alkoxycarbonyloxyalkyl radical with 3 to 14 carbon atoms, a dialkylaminoacetoxyalkyl radical with 4 to 21 carbon atoms, an alkanoylamidoalkyl radical with 2 to 13 carbon atoms, an aralkyl radical, where the alkyl portion has 1 to 3 carbon atoms and the aryl portion 6 to 10 carbon atoms, a mono- or bicyclic heteroaralkyl radical with 4 to 10 ring atoms and 1 to 6 carbon atoms in the alkyl moiety, where the Heteroatom an oxygen, sulfur or nitrogen atom 709822/1029 - 135 - 15833Y represents a ring-substituted aralkyl or heteroaralkyl radical » where the substituent is a chlorine, fluorine, bromine or iodine atom or a lower-alkyl radical with 1 to 6 carbon atoms, a lower alkanoyloxy radical having 1 to 6 carbon atoms, a lower alkoxy radical having 1 to 6 carbon atoms, a mono- or bicyclic heterocyclylalkyl radical, the heterocyclic Has 4 to 10 atoms and the heteroatom is an oxygen, sulfur or nitrogen atom and the Alkyl portion contains 1 to 6 carbon atoms, one Aryl or ring-substituted aryl radical with 6 to 10 ring carbon atoms, the core substituent being a Hydroxyl group, a lower alkyl radical having 1 to 6 carbon atoms, a chlorine, fluorine or bromine atom or a Is an alkylthioalkyl radical having 2 to 12 carbon atoms, a cycloalkylthioalkyl radical having 4 to 12 carbon atoms or an acylthioalkyl radical, where the acyl part has 2 to 10 carbon atoms and the alkyl part 1 to 6 Having carbon atoms means. 7. Compounds according to claim 6, characterized in that R is a radical of the following general formula: where X is an oxygen or sulfur atom, η 0 or 1, and R represents a hydrogen atom, an amino group or an alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, heteroaryl or heteroaralkyl radical,
or R is a radical of the general formula below (CH ₂ ) _n ZR 709822/1029 - 136 - 15833Y in which X is an oxygen or sulfur atom, R is a hydrogen atom, an amino, mercapto or hydroxyl group or an alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy, alkenyl, alkynyl, Aryl, aralkyl, acylcycloalkyl, heteroaryl or heteroaralkyl radical, η 0, 1, 2, 3 or 4 »
m 0 or 1 and Z are an oxygen, sulfur or nitrogen atom or a carbonyl group, with the proviso that if Z represents an oxygen atom, R is not a mercapto group or a hydrogen atom when Z is a sulfur atom R is not an amino or hydroxyl group, and when Z is a nitrogen atom, R is not a mercapto group is, or R is a radical of the general formula below is CHRR ¹
n where X is an oxygen or sulfur atom, η 0 or 1, R is a hydrogen atom, an amino, mercapto or hydroxyl group or an alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, Heteroaryl or heteroaralkyl ^{radical and R 1 is} an amino, hydroxy, azido, carbamoyl or guanidino group, an acyloxy radical, a halogen atom, a sulfamino, tetrazolyl, sulfo or carboxyl group, a carbalkoxy radical, a phosphono group or an alkoxy group Are alkylthio, with the proviso that not both radicals R and R ¹ can be a hydroxyl, amino or mercapto group at the same time, or
is: or R is a radical of the general formulas below 709822/102 9 - 137 - 15833Y ^{τ (0)} m ^(x) n (O) _n γ » where X is an oxygen or sulfur atom, a pure hydrogen atom, an amino, mercapto or hydroxyl group or an alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aralkoxy, alkenyl, alkynyl, aryl , Aralkyl, cycloalkyl, heteroaryl or heteroaralkyl radical, m and η are each independently O or 1, YO ^ M ^ -NR ₂ or R (where W ^ is a hydrogen atom, an alkali or alkaline earth metal cation or one derived from an organic base Represents a cation), and Y * and Y "are each independently 6 ^ mP _f -NR ₂ or R, where Y ¹ and Y" are also linked and together with the phosphorus atom to which they are bonded form cyclic esters or amides can, or R is a radical of the general formula below
is (CH ₂ ) _m -A- (CH ₂ ) _m -Y where m and η are each an integer from 0 to 5, ρ Ο
or T, A is an oxygen atom, NR '(R ^f is a hydrogen atom or a lower-alkyl radical with 1 to 6 carbon atoms), a sulfur atom or a single bond and Y represents one of the following radicals: 1) an amino group or substituted amino group of the general formulas -N (R) ₂ and where the radicals R, independently of one another, are each a hydrogen atom, N (R ¹ ) ₂ (R ¹ is a hydrogen atom or a lower alkyl radical having 1 to 6 carbon atoms) »a lower alkyl or lower alkoxy radical having 1 to 6 carbon atoms, a lower -alkoxy- 709822/1029 - 138 - lower-alkyl radical, the alkoxy portion having 1 to 6 carbon atoms and the alkyl portion 2 to 6 carbon atoms have »or a cycloalkyl or cycloalkylalkyl radical, the cycloalkyl moiety 3 to 6 carbon atoms and the alkyl moiety have 1 to 3 carbon atoms, where two R radicals be linked and together with the N atom to which they are bound, a ring with 3 to 6 atoms can form; 2) an amidino or substituted amidino group of the general formula where the radicals R, independently of one another, are each a hydrogen atom, N (R ') o ( ^Rl ^ ⁸ "^ a hydrogen atom or a lower-alkyl radical with 1 to 6 carbon atoms), a lower-alkyl or lower-alkoxy radical with 1 to 6 carbon atoms , a lower-alkoxy-lower-alkyl radical, the alkoxy part having 1 to 6 carbon atoms and the alkyl part having 2 to 6 carbon atoms (if the lower-alkoxy-lower-alkyl radical is bonded to a carbon atom, the alkyl part contains 1 to 6 carbon atoms), or a cycloalkyl or cycloalkylalkyl radical, where the alkyl moiety has 1 to 3 carbon atoms, two radicals R being linked and, together with the atoms to which they are bonded, can form a ring having 3 to 6 carbon atoms; 3) a guanidino or substituted guanidino group of the general formula 709822/1029 -139- 15833Y _Λ -NH-CN (R) ₉ NO where R has the meaning given under 2); 4) a guanyl or substituted guanyl group of the general formula -C = NR N (R) ₂ where R has the meaning given under 2); or 5) a nitrogen-containing, mono- or bicyclic heterocyclic (aromatic or non-aromatic) radical with 4 to 10 nucleus or ring atoms, with the heteroatom (s) except nitrogen (one) oxygen and / or sulfur atoms ) is (are). 8. Compounds according to claim 7, characterized in that X in the radical COXR is an oxygen atom and R is Hydrogen atom, a methyl, tert-butyl, phenacyl, p-bromophenacyl, pivaloyloxymethyl, 2,2,2-trichloroethyl, Allyl, 3-methyl-2-butenyl, 2-methyl-2-propenyl, benzyl, benzylhydryl, p-tert-butylbenzyl, Phthalidyl, phenyl, 5-indanyl, acetylthiomethyl, Acetoxyraethyl, propionyloxymethyl, methallyl, 3-butenyl, 4-pentenyl, 2-butenyl, acetoxyacetylmethyl, pivaloylacetylmethyl, diethylaraino, dirnethylaminoethyl, Methoxymethyl, p-acetoxybenzyl, p-pivaloylbenzyl, p-Isopropoxybenzyl, 5-indanylmethyl, benzyloxymethyl, Methylthioethyl, dimethylaminoacetoxyme-. ethyl, crotonolacton-3-yl, acetamidomethyl, acetylthioethyl, Represents pivaloylthiomethyl or methylthiomethyl group. 709822/1029 9. Compounds according to claim 8, characterized in that the radical R has the general formula below in which R is a benzyl, p-hydroxybenzyl, 4-amino-4-carboxybutyl, Methyl, cyanine thyl, 2-pentenyl, n-amyl, n-heptyl, ethyl, 3- or 4-nitrobenzyl, Phenethyl, β, β-diphenylethyl, methyldiphenylmethyl, Triphenylmethyl-, 2-methoxyphenyl-, 2,6-dimethoxyphenyl-, 2,4 »6-trimethoxyphenyl-, 3» S-dimethyl ^ -isoxazolyl-, 3-butyl-5-methyl-4-isoxazolyl-, 5-methyl-3-ph.enyl-4-isoxazolyl-, 3- (2-chlorophenyl) -5-meth.yl-4-isoxazolyl-, 3- (2,6-dichlorophenyl) -5-meth.yl-4-isoxazolyl-, D-4-amino-4-carboxybutyl-, D-4-N-Benzoylamino-4-carboxy-n-butyl-, p-aminobenzyl-, o-aminobenzyl-, m-aminobenzyl-, p-Dimethylaminobenzyl-, (3-pyridyl) -methyl-, 2-ethoxy-1-naphthyl-, 3-Carboxy-2-quinoxalinyl-, 3- (2,6-I) chlorophenyl) -5- (2-furyl) -4-isoxazolyl-, 3-PhOHyI ^ -IsOXaZoIyI-5-methyl-3- (4- guanidinophenyl) -4-isoxazolyl-, 4-guanidinomethylphenyl-, 4-guanidinomethylbenzyl-, 4-guanidinobenzyl-, 4-guanidinophenyl-, 2,6-dimethoxy-4-guanidinophenyl-, o-sulfobenzyl, p-carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-naphthylmethyl, 3-isothiazolylmethyl, 4-isothiazolylmethyl-, 5-isothiazolylmethyl-, Guanylthiomethyl, 4-pyridylmethyl, 5-isoxazolylmethyl, 4-methoxy-5-isoxazolylmethyl-, 4-methyl-5-isoxazolylmethyl-, 1-imidazolylmethyl-, 2-benzofuranylmethyl-, 2-Indolylmeth.yl-, 2-Ph.enylvinyl-, 2-Ph.enyläthinyl-, I-aminocyclohexyl-, 2- or 3-thlenylaminomethyl-, 2- (5-nitrofuranyl) vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl-, o-phenylphenyl-, p-aminomethylbenzyl-, 1- (5-cyantriazolyl) methyl, difluoro 709822/1029 methyl-, dichlorometh.yl-, dibromomethyl-, 1- (3-methylimidazolyl) -methyl-, 2- or 3- (5-carboxymethylthienyl) methyl-, 2- or 3- (4-carbamoylthienyl) -methyl-, 2- or 3- (5-methylthienyl) -methyl-, 2- or 3- (5-methoxythienyl) -methyl-, 2- or 3- (4-chlorothienyl) methyl, 2- or 3- (5-sulfothienyl) methyl, 2- or 3- (5-carboxythienyl) methyl, 3- (i, 2,5-thiadiazolyl) methyl, 3- (4-methoxy-1,2,5-thiadiazolyl) methyl, 2-purylmethyl-, 2- (5-nitrofuryl) -methyl-, 3-purylmethyl-, 2-thienylmethyl, 3-thienylmethyl, tetrazolylmethyl, Means benzamidinomethyl or cyclohexylamidinomethyl group. 10. Compounds according to claim 8, characterized in that the radical R has the general formula below (CH ₂ ) _n ZR where the portion - (CH ₂ ) _Q ZR of the acyl radical is an allylthiomethyl, phenylthiomethyl, butyl mercaptomethyl, α-chlorocrotyl mercaptomethyl, phenoxymethyl, phenoxyethyl, phenoxybutyl, phenoxybenzyl, diphenoxymethyl, dimethyl methoxymethyl, dimethyl methoxymethyl phen -, 4-G-uanidinophenoxymethyl-, 4-pyridylthiomethyl-, p- (carboxymethyl) -phenoxymethyl- »p- (carboxymethyl) -phenylthiomethyl-, 2-thiazolylthiomethyl-, p- (sulfo) -phenoxymethyl-, p- (carboxymethyl ) -phenylthiomethyl-, 2-pyrimidinylthiomethyl-, phenethylthiomethyl-, 1- (5 »6,7» 8-tetrahydronaphthyl) -oxymethyl-, N-methyl-4-pyridiniumthio-, benzyloxy-, methoxy-, ethoxy-t Phenoxy, phenylthio, amino, methylamino, dimethylamino, pyridiniummethyl, trimethylammoniummethyl, cyanomethylthiomethyl, trifluoromethylthiomethyl, 4- 7 0 9 8 2 2/1029 - 142 - Pyridylethyl, 4-pyridylpropyl, 4-pyridylbutyl, 3-imidazolylethyl, 3-imidazolylpropyl, 3-imidazolyl- ■ butyl, 1-pyrroloethyl, 1-pyrrolopropyl or 1-pyrrolobutyl group represents. 11. Compounds according to claim 8, characterized in that the radical R has the following general formula HRR ¹ where the -CHRR * portion of the acyl radical is an a-aminobenzyl- » a-amino- (2-thienyl) -methyl-, a- (methylamino) -benzyl-, α-aminomethyl mercaptopropyl, α-amino-3 or -4-chlorobenzyl-f a-amino-3- or -4-hydroxybenzyl- » a-amino-2,4-dichlorobenzyl- »a-amino-3» 4-dichlorobenzyl-, D - (-) - a-Hydroxybenzyl-, a-Carboxybenzyl-, a-Amino- (3-thienyl) -methyl- » D - (-) - a-amino-3-chloro-4-hydroxybenzyl-, a-amino- (cyclohexyl) -methyl- »a- (5-tetrazolyl) -benzyl-, 2-thienylcarboxymethyl-, 3-thienylcarboxymethyl- » 2-furylcarboxymethyl-, 3-furylcarboxymethyl-t α-sulfaminobenzyl-, 3-thienylsulfaminomethyl-, a- (N-methylsulfamino) -benzyl-, D - (-) - 2-thienylguanidinomethyl-, D - (-) - a-guanidinobenzyl-, a- & uanylureidobenzyl- » a-hydroxybenzyl- »a-azidobenzyl-, a-fluorobenzyl-» 4- (5-methoxy-1,3-oxadiazolyl) aminomethyl » 4- (5-methoxy-1 »3-oxadiazolyl) -hydroxymethyl-, 4- (5-methoxy-1» 3-bulfadiazolyl) -hydroxymethyl-, 4- (5-chlorothienyl) aminomethyl-, 2- (5-chlorothienyl) -hydroxymethyl-, 2- (5-chlorothienyl) -carboxymethyl-, 3- (i »2-thiazolyl) -aminomethyl-, 3- (i »2-thiazolyl) -hydroxymethyl-, 3- (i» 2-thiazolyl) -carboxymethyl-, 2-thiazolylaminomethyl-, 2-thiazolylhydroxymethyl- »2-thiazolylcarboxymethyl-» 2-benzothienylcarboxymethyl-, 2-benzothienylhydroxymethyl-, 2-Benzothienylcarboxymethyl-, α-sulfobenzyl-t a-phosphonobenzyl, a-diethylphosphono or a-monoethylphosphono group represents. 709822/1029 ^15833Ύ Μ 2652675 12. Compounds according to claim 8, characterized in that that the radical R is one of the following groups O NH .0 NH I! Il Il Il -CCH ₂ CH ₂ NHC-CH ₃ , -CCH ₂ CH ₂ CH ₂ NHc-CH ₃ ? T H -CCH ₂ CH ₂ NHC-H, 0 NH ο ii -CCH ₂ CH ₂ NHC-NH ₂ , I? ^ -CCIUCIUN (CH.) ". -CCH ₂ SC _n * _o v ^ ii _o ιί \ «_ n— / - 2 \ Φ β -CCH ₀ CH ₀ R (CH-),, Π ^{22 J 3} -CCH ₂ S-CH ₂ -C Ο NH -CCH ₂ CH ₂ C-NH ₂ , 9 -CCH ₀ -O-CH ₀ C ^{2 2} 13 · Compounds according to claim 8, characterized in that R ^ is a pormyl, propionyl, butyryl, chloroacetyl, Methoxyacetyl, aminoacetyl, methoxycarbonyl, ethoxycarbonyl, Methylcarbamoyl, ethylcarbamoyl, phenylthiocarbonyl, 3-aminopropionyl, 4-aminobutyryl, N-methylaminoacetyl, NjN-dimethylaminoacetyl-, N, N, N-trimethylaminoacetyl-, 3- (N, N-dimethyl) -aminopropionyl-, 3- (N, N, N-trimethyl) -aminopropionyl-, N, N, N-triethylaminoacetyl-, Pyridinium acetyl, guanylthioacetyl, gunaidinoacetyl, 3-guanidinopropionyl-, N -methylguanidinopropionyl-, Hydroxyacetyl, 3-hydroxypropionyl, acryloyl, Propinoyl, malonyl, phenoxycarbonyl, amidinoacetyl, Acetamidinoacetyl, Amidinopropionyl, Acetamidinopro- 709822/10 29 pionyl, guanylureidoacetyl, guanylcarbamoyl, carboxymethylaminoacetyl, Sulfoacetylaminoacetyl, phosphono ac etylamino ac etyl-, N -Dimethylamino ac etamidinopropionyl-, Ureidocarbonyl, dimethylaminoguanylthioacetyl, 3- (1-methyl-4-pyridinium) propionyl, 3- (5- aminoimidazo1-1-yl) propionyl, 3-methyl-1-imidazolium acetyl, 3-sydnonylacetyl-, o-aminomethylbenzoyl- or ο-aminobenzoyl group or one of the following radicals ff SB ^Q Il Il 11 ^0CH 3 -P (OCH,) ,,, -P (OCH,) 2, -Ρ / ', ^{02 J z} OM S S -PN (CH ₃ J ₂ '0M ( ₃ J ₂ , [( ₃₂₂ OM where M is a hydrogen atom or an alkali or alkaline earth metal cation represents, means. 14. Compounds according to claim 8, characterized in that that R is a sulfo, phosphono, carbamoyl, methylsulfonyl, Sulfamoyl, dirnethylsulfamoyl, N-methylcarbamoyl, Bromoacetyl, hydroxyacetyl, aminoacetyl, dime thylamino acetyl, trimethylammonium acetyl, amidinoacetyl, Guanidinoacetyl, methoxyacetyl, guanylacetyl, Guanylthioacetyl, phosphamoyl, phosphonothioyl, thiocarbamoyl, Methoxymethyl, hydroxyethyl, methoxyethyl, Dimethylaminomethyl, dimethylaminoethyl, methylthiomethyl, Represents amidinomethyl or guanidinoethyl group, 709822/1029 15833Y ^ .... 15. Compounds according to claim 6, characterized in that that χ is an oxygen atom, O R ^{4 is} a hydrogen atom, CH ^, SO-Ji, CH ₉ OCHo, COCH ^ or O ό 1 d S 5 CCH ₂ NH ₂ , O R is a hydrogen atom, CH ₂ OCCH ₃ , CH ₂ OCH ₂ C (CH ₃ ) ₃ , CH ₂ CH = CH ₂ , CH ₃
CH ₂ C = CH ₂ , CH ₂ CH ₂ N (C ₂ H ₅ ) ₂ , CH ₂ - / ~ \ -NO ₂ , CH ₂ CH ₂ CH ₃ , CH ₂ CH ₂ N (CH ₃ ) ₂ , CH ₂ C - / ^, CH ₂ SCH ₃ or CH ₂ OC (CH ₃ J ₃ 5 6 7 and R, R and R are each independently a hydrogen atom (not all radicals R, R and R are hydrogen atoms at the same time), CH ₃ , CH ₂ CH ₃ , CH ₂ CH = CH ₂ , 'CH ^ CHoCHt, CHp (C ₆ H ₁ -), CH (C ₆ Hp) P, C (CgH ₅ ) ₃ , CH ₂ C = CH ₂ , CH ₂ OCH ₃ , CH ₂ CH = CH-CH ₃ CH ₃ I NH CH ₂ CK ₂ CH = CH ₂ , ^CH 2 ~ \ /) " -CH ₂ CH ₂ CH ₂ ", ■ - + 9 CH ₂ CH ₂ N (CH ₃ ) ₃ / CH ₂ CCH ₃ , CH ₂ CH ₂ CH ₂ -C = CH, -CH ₂ CH ₂ OCH ₂ CH ₂ "", CH ₂ C CH ₂ SCH ₃ , CH ₂ OC (CH ₃ ) ₃ , CH ₂ CH CH ₂ CH ₂ COCH ₃ , · / ~ \ NO ₂ NO ₂ CH (CH ₃ ) ₂ , CH ₂ CH ₂ CH ₂ CH ₃ , CH _: CH ₃ CHCH ₂ CH _3,. CHCH ₂ CH ₂ CH ₃ , CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ , CH ₃ CH. 709822/1029 15833Y Jib CH ₂ CHCH ₂ CH ₃ , CH ₂ CH ₂ CHCH ₃ , CH. CH. CH CH CH- CH, CH, CH ₂ C (CH ₃ J ₃ , CHCH ₂ CH (CH ₃ ) CH., W / r CH. CH ₂ CH = CH. / CH ₂ CH = CH OCH- mean. 16. Compounds according to claim 15, characterized in that R is a hydrogen atom, X is an oxygen atom and R mean a hydrogen atom, as well as their pharmacologically acceptable salts. 17. Process for the preparation of the compounds according to claim 6, characterized in that a compound the general formula below OR ⁴ Jl- COXR or an N-alkyl derivative of such a compound with ^ f \ brings an alkylating agent suitable for introducing the substituents R, R and R into reaction. 709822/1029 - 147 - 18. Process for the preparation of the compounds according to claim 6, characterized in that a compound the general formula below fl 8 SCH ₂ CH ₂ NHCR COXR in which R is a hydrogen atom, a lower alkyl radical having 1 to 5 carbon atoms or an aralkyl radical with 6 to 10 ring carbon atoms and 1 to 5 carbon atoms in the alkyl chain means hydrogenated in the presence of a catalyst. 19 · Medicines characterized by a content of one Connection according to claim 1 in addition to the usual auxiliary and / or packaging means. 20. Medicament containing, in single dosage form, a therapeutically effective amount of a compound according to claim 1 in addition to the usual auxiliary and / or packaging materials. 21. Medicines, characterized by a content of a compound according to claim 6 in addition to the usual auxiliary and / or packaging means. 22. Medicament containing, in unit dosage form, a therapeutically effective amount of a compound according to claim 6 in addition to the usual auxiliary and / or packaging materials. 709822/1029