SK278319B6 - (z)-2-acylamino-3-monosubstituted propenoates, method of their manufacture and antibacterial drugs containing them - Google Patents

Abstract

The compounds called (Z)-2-acylamino-3-mono substituted propionates with a general formula (I), where a meaning of the substituents is explained in the description, are the dipeptidase inhibitors which can be used in the antibacterial preparations made based on the antibiotics closed to the group of thienamycine. This patent contains a description closed to the procedures dealing with the production of these compounds.

Description

Technical field

The invention relates to (Z) -2-acylamino-3-monosubstituted propenoates which are effective as dipeptidase inhibitors, processes for their preparation and antibacterial preparations containing a combination of these substances with antibiotics of the thienamycin family.

BACKGROUND OF THE INVENTION

Recently, a new group of condensed β-lactam ring antibiotics including thienamycin and its semi-synthetic derivatives, epitenamycins and olivanoic acids has been described. These compounds will be described in more detail below and will be referred to as the thienamycin group. These substances have a high antibacterial activity, but are very rapidly metabolized in the mammalian organism.

Metabolism occurs mainly in the kidneys and an enzyme has been obtained from the kidneys to catalyze the inactivation of thienamycin by hydrolysis of the β-lactam ring. The enzyme was then localized cytologically and examined for substrate specificity and inhibition, and found to be very similar to renal dipeptidase (EC 3.4.12.11), which is also called dehydropeptidase-I in the literature, but it is not identical to this enzyme. β-Lactamase activity can only be demonstrated with respect to the thienamycin group. So far, there are no known examples of metabolism associated with β-lactam ring cleavage in mammals for well-known β-lactam type antibiotics such as penicillins and cephalosporins.

Chemicals that selectively inhibit the metabolism of dipeptidase (E. C. 3.4.1.11) are called dipeptidase inhibitors.

the oxidized form of the sulfur, and the terminal hydrogen atom can be replaced by a hydroxyl or thiol group, or an amino group which is optionally derivatized in the form of 1 to 2 carbon acylamino, ureido, amidine, guanidine, monoalkylamino, dialkylamino and trialkylamino groups. up to 2 carbon atoms in each of the alkyl moieties, followed by carboxy, a group of formula (HO / 2P / O) - or - SO ₃ OH, or an appropriate alkyl ester or alkylamide group having 1 to 2 carbon atoms in the alkyl moiety derived from any of these acids , cyano or combinations thereof, such as a suitable terminal amino acid residue, wherein when R ³ is C 1 -C 4 straight chain alkyl, R ^{2 is not} C 1 -C 4 straight chain alkyl.

The present invention further provides a process for the preparation of compounds of formula (I) wherein R ¹ is hydrogen, C 1 -C 6 alkyl or a pharmaceutically acceptable cation and R ² and R ³ are as defined in claim 1, according to claim 1, which consists of up to 4 parts by weight of the 2-keto acid of the general formula

H, where R ^j CH ₂ --C ₂ h wherein R ³ is as defined in Claim 1, or an alkyl ester having 1 to 6 carbon atoms in the alkyl radical condensed with 1 part of an amide of formula

O r ² cnh ₂

SUMMARY OF THE INVENTION

The present invention provides (Z) -2-acylamino-3-monosubstituted propenoates of formula (I)

R ³ 11 \ /

C

II / 1 / where

R ¹ is hydrogen, C 1 -C 6 alkyl, di (C 1-2 alkyl) amino (C 1-3 alkyl) or a pharmaceutically acceptable cation;

R ² is a hydrocarbon radical of 3 to 10 carbon atoms in which the carbon chain may be from 1 to 6 hydrogen atoms are replaced by halogen atoms, and a methylene group in the 60 non-terminal position can be replaced by oxygen or sulfur, including oxidized forms of sulfur;

R ³ represents a hydrocarbon radical having 1 to 15 carbon atoms, in which the carbon chain may be 1 to 6 65 carbon atoms replaced by halogen atoms, the methylene group other than the terminal position may be replaced by an oxygen or sulfur atom, including where R ² is as defined in claim 1, by refluxing the reaction mixture in an inert solvent such as toluene or methylisovalerate while simultaneously azeotropically separating water, then the optionally formed free acid of formula (I) is converted to its C 1 -C 6 alkyl ester, or the resulting ester is converted to the free acid, or the resulting product is converted to its pharmaceutically acceptable salt.

The present invention also provides a process for the preparation of compounds of formula (I) according to claim 1, which comprises reacting an acid chloride of the formula

Where R ² is as defined above, with tert-butyl ester of an amino acid of the general formula

where

R ³ is as defined above, in the presence of a base followed by oxidative addition of sodium methoxide, and the resulting product is treated with anhydrous hydrogen chloride, then optionally the resulting product is converted to its pharmaceutically acceptable acid addition salt or other ester within the definition of R ¹ .

Finally, the invention also provides an antibacterial preparation based on an antibiotic of the thienamycin family, such as thienamycin, N-formimidoylthienamycin and N-acetylimidoylthienamycin, comprising combining the antibiotic with a dipeptidase inhibitor of formula (I) according to claim 1, in a weight ratio of the first agent to the second agent in the range of 1: 3 to 30: 1.

Thus, the terminal hydrogen atom in R ³ may also be replaced by a hydroxyl group or a thiol group, which may be acylated, for example, with a C 1 -C 8 alkanoic acid or carbamoylated, including alkyl and dialkyl carbamate derivatives. The hydrogen atom may also be replaced by an amino group which may be substituted and thus may be an acylamino group, an ureido group, an amidine group, a guanidine group or an amino group substituted with an alkyl group, optionally further substituted, including quaternary ammonium groups. It may furthermore be a substitution by acidic groups, for example by carboxylic, phosphonic or sulphonic acid residues, or by esters or amides of these acids, or by a cyano group, or a combination of these substituents, in particular in the case of terminal amino groups.

The substituent ^R2 is preferably a branched alkyl or cycloalkyl of 3 to 10 carbon atoms with the exception of the groups in which a carbon atom that is located immediately adjacent the carbonyl group is tertiary. In addition, when R ³ is C 1 -C 4 straight chain alkyl, R ^{2 is} different from C 1 -C 4 alkyl. R ¹ represents a hydrogen atom, a C 1-6 alkyl or a dialkylaminoalkyl group, for example -CH 2 CH 2 N / C 2 H 5/2 groups. -CH 2 CH / CH 3 / N / CH3 / second

Some compounds of formula (Π) are asymmetric. The racemic (Z) - (2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid) was resolved and found to be bound to the dextrorotatory isomer having the S-configuration.

The substituent R ² can be distinguished the following groups:

-R ⁴ (IA) wherein

R ⁴ is a straight chained, branched or cyclic chain hydrocarbon radical having from 3 to 10 carbon atoms, optionally substituted as described herein;

-R ⁵ R ⁶ (IB) wherein

R ⁵ represents cycloalkyl of 3 to 6 carbon atoms,

R ⁶ represents one or two alkyl groups optionally linked and forming a further ring on the cycloalkyl ring, which substituents may be further substituted as described above,

-R ⁷ R ⁸ (IC) wherein

R 'represents an alkylene group having 1 to 3 carbon atoms and

R ⁸ represents C 3 -C 6 cycloalkyl, which substituents may be further substituted as described above.

The following are compounds that fall within the scope of each subgroup:

IA:

(Z) -2-isovaleramido-2-pentenoic acid, (Z) -2-isovaleramido-2-butenoate, (Z) -2-isovaleramido-2-butenoic acid, (Z) -2-benzamido-2-butenoic acid (Z) -2- (3,5,5-trimethylhexanamido) -2-butenoic acid, (Z) -2-cyclobutanecarboxamido-2-butenoic acid, (Z) -2-cyclopropanecarboxamido-2-butenoic acid, ( Z) -2-cyclopropanecarboxamido-2-pentenoic acid, (Z) -2- (3-methylvaleramido) -2-butenoic acid, (Z) -2-cycloheptanecarboxamido-2-butenoic acid, (Z) -2-nonanamido- 2-butenic acid, (Z) -2-cyclohexanecarboxamido-2-butenoic acid, (Z) -2- (4-methylvaleramido) -2-butenoic acid, (Z) -2-tert-butylacetamido-2-butenoic acid, (Z) -2-octanamido-2-butenoic acid, (Z) -2-butyramido-2-butenoic acid, (Z) -2-valeramido-2-butenoic acid, (Z) -2-cyclopentanecarboxamido-2-butenoic acid (Z) -2- (6-methylheptanamido) -2-butenoic acid, (Z) -2-hexanamido-2-butenoic acid, (Z) -2- (3,7-dimethyloctanamido) -2-butenoic acid, acid (Z) -2- (3,7-dimethyl-6-octenamido) -2-butenoic acid (Z) -2- (5-chloro-valeramido) -2-butenoic acid (Z) -2- (3-chlorobenzoylamido) -2 -butenic acid, (Z) -2- (2-chlorobenzamido) -2-butenoic acid, (Z) -2-nonanamido-2-butenoic acid.

(Z) -2- (6-bromohexanamido) -2-butenoic acid, (Z) -2- (3,3-dimethylpropenamido) -2-butenoic acid, (Z) -2-benzamido-2-cinnamic acid, acid (Z) -2-benzamido-2-pentenoic acid, (Z) -2-benzamido-5-methoxy-2-pentenoic acid, (Z) -2-benzamido-2-hexenedioic acid, (Z) -2-isovaleramido -2-octenoic acid, (Z) -2-isovaleramido-2-cinnamic acid, (Z) -2-isovaleramido-2-hexenedioic acid, (Z) -2-cyclopropanecarboxamido-2-cinnamic acid, (Z) -2 -cyclopropanecarboxamido-2-hexenedioic acid, (Z) -2- (5-methoxy-3-methylvaleramido) -2-butenoic acid, (Z) -2-ethylthioacetamido-2-butenoic acid, (Z) -2- (2) (2-Dichlorocyclopropanecarboxamido) -2-butenoic acid (Z) -2- (2-ethylhexanamido) -2-butenoic acid, (Z) -2-di-n-propylacetamido-2-butenoic acid,

IB:

(Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoic acid, (+) - Z-2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoic acid, (Z) -2- (2, 2-dimethylcyclopropanecarboxamido) -2-pentenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-hexenoic acid, (Z) ) -2- (2,2-dimethylcyclopropanecarboxamido) -2-cinnamic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -5-methoxy-2-pentenoic acid, (Z) -2- (2,2) (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -3- (2-chlorophenyl) propenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -dimethylcyclopropanecarboxamido-4,4-tritluoro-2-butenoic acid -dimethylcyclopropanecarboxamido) -2-hexenedioic acid, (Z) -2- (2-ethylcyclopropanecarboxamido) -2-butenoic acid, (Z) -2- (2,2-diethylcyclopropanecarboxamido) -2-butenoic acid, (Z) -2 - (2 _> 2-diethylcyclopropanecarboxamido) -2-pentenoic acid (Z) -2- (2-isopropyl-2-methylcyclopropanecarboxamido) -2-butenoic acid (Z) -2- (2-methyl) (cyclohexanecarboxamido) -2-butenoic acid (Z) -5-cyano-2- (2,2-dimethylcyclopropanecarboxainido) -2-pentenoic acid.

(Z) -5- (N, N-dimethylcarbamoyl) -2- (2,2-dimethylcyclopropanecarboxamido) -2-pentenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -5-methanesulfonyl-2- pentenoic acid (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -5-ethoxycarbonyl-2-pentenoic acid (Z) -2- (2-methylcyclopropanecarboxamido) -2-butenoic acid methyl- (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoate, ethyl (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoate,

(Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoic acid, 2-dimethylaminoethyl ester,

(Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-pentenoic acid, 3-diethylaminopropyl ester, (Z) -2- (2,3-dimethylcyclopropanecarboxamido) -2-butenoic acid, (Z) -2- ( 3,3-dimethylcyclobutanecarboxamido) -2-butenoic acid, (Z) -2- (2-spirocyclopentanecarboxamido) -2-butenoic acid, (Z) -2- (2-tert-butyl-3,3-dimethylcyclopropanecarboxamido) -2-butenoic acid (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -4-methyl-2-pentenoic acid, (Z) -2- (2-tert-butylcyclopropanecarboxainido) -2-butenoic acid, (Z) -2- (2-phenylcyclopropanecarboxamido) -2-butenoic acid, (Z) -3-cyclohexyl-2- (2,2-dimethylcyclopropanecarboxamido) propenoic acid, (Z) -5-carboxy-5- (2,2-dimethylcyclopropanecarboxamido) -4-pentenamidine, (Z) -5-dimethylamino-2- (2,2-dimethylcyclopropanecarboxamido) -2-pentenoic acid, (Z) -3-cyclopropyl-2- (2,2-dimethylcyclopropanecarboxamido) propenoic acid, (Z) -2- (2) (2-dimethylcyclopropanecarboxamido) -2,5-hexadienoic acid (Z) -2,1,2,2-dimethylcyclopropanecarboxamides do) A-phenyl-2-butenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -6-mercapto-2-hexenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -5- methylthio-2-pentenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -5-phosphono-2-pentenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) 2-heptenoic acid, (Z) ) -2- (2,2-dimethylcyclopropanecarboxainido) -5-phenyl-2-pentenoic acid (Z) -2- (2,2-dimethylcyclopropanecarboxarnido) -2-nonenoic acid (Zj-2- (2,2) -dimethylcyclopropanecarboxamido) -2-decenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-tridecanoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -6-methoxy-2-hexenoic acid, and 5-Methoxy-2-pentenoic acid (Z) -2- (2,2-dimethylcyclopropanecarboxamido) 15 -6-methyl-2-heptenoic acid (Z) -4-cyclohexyl-2- (2,2-dimethylcyclopropanecarboxamido) 2-butenoic acid

IC:

2θ (Z) -2-cyclobutylacetamido-2-butenoic acid, (Z) -2-cyclopentylacetamido-2-butenoic acid, (Z) -2-cyclohexylacetamido-2-butenoic acid, (Z) -2- (4-) (cyclohexylbutyramido) -2-butenoic acid, (Z) -2-cyclopropylacetamido-2-butenoic acid,

(Z) -2-cyclopropylacetamido-2-pentenoic acid, (Z) -2- (3-cyclopentylpropionamido) -2-butenoic acid, (Z) -2- (3-cyclohexylpropionamido) -2-butenoic acid (Z) -2- [4- (2-thienyl) butyramido] -2-butenoic acid (Z) -2- (4-phenylbutyramido) -2-butene (D, La-lipoamido) -2-pentenoic acid (Z) -2- (D, La-lipoamido) -2-cinnamic acid, (Z) -2- [3- (2-tetrahydrofiiryl) propionamido] -235-butenoic acid.

Particularly preferred substituents for R ² are 2,2-and 2,2-dimetylcyklopropylová dichlorocyclopropyl group.

Particularly preferred R ³ groups are

N-alkyl groups in compounds having 1 to 9 carbon atoms, in particular N-methyl groups in compounds having 1 to 9 carbon atoms with a terminal substituent which is a quaternary nitrogen atom, an amine derivative or an amino acid group.

The term quaternary nitrogen atom means a tetrasubstituted or heteroaromatic positive nitrogen atom. Otherwise, it may be an ammonium group substituted with hydrocarbon groups having 1 to 7 carbon atoms, which may be the same or 50 different.

The term "amine derivative" refers to amino, acylamino, ureido, amidine, guanidine and alkyl derivatives of these groups.

An amino acid group refers to, for example, a cysteinyl group (-SCH 3 CH) - (NH 2) COOH or a sarcosyl group -N (CH ₃ ) CH 2 -COOH. in which the hydrogen bonded to an oxygen, nitrogen or sulfur atom in the known 60 amino acids has been replaced.

Particularly preferred compounds are those wherein R ² is 2,2-dimethylcyclopropyl or 2,2-dichlorocyclopropyl and R ³ is C 3-7 hydrocarbon radical without or without a terminal substituent which is a trimethylammonium group , amidino, guanidino, 2-amino-24 carboxyethylthio or ureido. Specific examples of preferred compounds will be given below: (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-trimethylanionium-2-octenoic acid inner salt, (Z) -2- (2,2-dichlorocyclopropanecarboxamido) inner salt - 8-trimethylammonium-2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-amidino-2-octenic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-guanidino- 2-Octenoic acid (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-ureido-2-octenoic acid (Z) -8- (L-2-amino-2-carboxyethylthio) -2- (2) , 2-dimethylcyclopropanecarboxamido) -2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid (racemic and dextrorotatory form) and (Z) -2- (2,2-dichlorocyclopropanecarboxamido) - 2-octenoic.

The compounds Z were assigned the configuration Z (JE Blackwood et al., J. Am. Chem. Soc. 90, p. 509 (1968) based on their NMR spectrum analogously to A. Srinavasan et al., Tetrahedron Lett., 891 (1976) ).

Although these compounds of formula (I) in which R ¹ is a hydrogen atom are described and named as free acids, it is well understood that pharmaceutically acceptable derivatives of these acids, such as salts, with alkali metals, alkaline earth metal salts, ammonium salts or amine salts. Suitable salts are, for example, sodium, potassium, calcium or tetramethylammonium salts.

As already mentioned, the compounds produced by the process of the invention are dipeptidase inhibitors (E. C. 3.4.1.13) and can be used in admixture with antibacterial compounds that are subject to renal degradation. A group of antibiotics of fundamental importance for this use together with the (Z) -2-acylamino-3-monosubstituted propenoates of the invention are thienamycin compounds.

Compounds of the thienamycin family refer to a large number of naturally occurring, semisynthetic or synthetic derivatives or analogs thereof having a β-lactam ring with another fused ring. These compounds can be considered as 6- or 2-substituted 2-penem-3-carboxylic acids and 1-carbodetia-2-penem-3-carboxylic acids or 1-azabicyclo [3,2,2] -2-heptene-7 -one-2-carboxylic acids.

Specific compounds particularly useful for this purpose can be represented by the general formula (Π)

where

X represents a methylene group or a sulfur atom, R ² represents a hydrogen atom, -S-CH ₂ CH ₂ NH ³ , where

R ³ represents a hydrogen atom, an acetyl group, a formimidoyl group, a acetimidoylovú, -S (O) -CH = CHN H CO _3, or -S-CH = CHN H CO _3, and

R ⁶ represents a hydrogen atom or a group - CHCH ₃ ,

R ⁷ where

R ⁷ represents a hydrogen atom, a hydroxy group or a sulfonyloxy group.

All stereoisomeric forms are also included in the structural formula.

All compounds of formula (H) are described in the literature. When X is methylene and R ² is SCII ² Cl ² NH 2, and R ⁶ is CH / OH / CH 3, it is thienamycin, an antibiotic produced by S. cettley, described in U.S. Pat. The fermentation product N-acetylthioenamycin in which R ⁵ is a CH / OH / CH 3 group and R ³ is acetyl is a product called 924A of Belgian patent no. No. 848,346, issued May 16, 1977. The N-imidoyl derivative is disclosed in Belgian patent no. No. 848,545 of May 20, 1977. Compounds with an unsaturated side chain, called N-acetyldehydrothienamycin and 924Aj, are fermentation products described in US SN 788 491, filed April 18

The epimeric form of N-acetylthienamycin, also called 890A and 890A ₃ , as well as desacetyl 890A and desacetyl 890A ₃ , was described in French application 763 887, filed On Nov. 19, 1976 and in US SN 634,300, filed on Nov. 21, 1975 as Time 15745; 848 349 of 16 May 1977, the epimeric forms of the unsaturated thienamycin, also called 890A ₂ and 890Ä5, are described in French patent application Laid-28 April 1976, the 15,839th Time of N-acetylated compounds containing a 6-sulfonyloxy, called 890A ₉ and 89OAio, are described in French patent no. No. 7,734,456 of Nov. 16, 1977, and US Priority Document of Nov. 17, 1976; Time 15935; No. 7,734,457 of Nov. 16, 1977 with US Priority Document of Nov. 17, 1976, Time 15936. The desacetylanalogues of compounds 890A ₉ and 89OAio are described in US SN 860 665, filed on Feb. 11, 1977 as Time 15975 and fallen in its continuation of US SN 850 665, filed December 15, 1977, as well as French Patent 7,803,666 of February 9, 1977;

1978 and US SN 767 920, filed February 11, 1977 as Time 15976 and fell, as well as its sequel UC SN 006 959, filed January 25, 1977.

1979 and in French patent no. 7,803,667, which was brought on 9 February 1978. Some of the compounds, primarily derivatives of 890A ₉ and 890A] _0, are also known as derivatives of olivanovej and are described in Corbett et al, J. Chem. Soc. Chem. Commun. 1977, no. 24, pages 953 to 954. The compounds of formula (I) wherein R ² is hydrogen and which is also called the dezcysteaminyltienamycíny described in U.S. SN 668,898 of 22 March 1976 Time of 15866 that fell, and its continuation U.S. SN No. 847,297, filed October 31, 1977, and fell, as well as in Belgian patent no. 867 227 of 20 November 1978.

Compounds in which R ⁶ is a hydrogen atom and X is a methylene group have been described in US SN 843 171, filed January 1, 1977 as Time 15902, and in U.S. Pat. No. 2,751,624 1, 18 November 1977.

An antibiotic of the thienamycin family wherein R ² is -SCH ₂ CH ₂ NHAc and R ⁶ is an ethyl moiety has been described under the name PS-5 by K. Okaimura et al., J. Antibiotics 31, p. 480 (1978) and Belgian patent no. 865 578.

Compounds in which X is a sulfur atom and which are also described as foam compounds have been reported in R. B Woodward Recent Advances in the Chemistry of β-Lactam Antibiotics, J. Alks (Ed), The Chemical Society, London, 1977 , p. 167, R. B. Woodward, Abstracts of Uppsala University 500 Years Symposium on Current Topics in Drug Reasearch, Uppsala, 10 Sweden, October, p. 1921 (1977) Acta. Pharm. Suecica, Vol. 14, Supplement, p. 23, and U.S. Pat. No. 4,070,477 of January 24, 1978.

Particularly preferred compounds of the thienamycin family are N-formimidoyl and N-acetamidoyl derivatives of thienamycin. The crystalline form of N-formimidoylthienamycin, described recently, is also useful. The preferred method of making the compound will be described below.

Process for producing crystalline N-formimidoylthienamycin

Step A - Benzylformimidate hydrochloride

A 3 liter three-necked flask with funnel, stirrer and reflux condenser was charged with a mixture of 25,125 g (1.15 mol) benzyl alcohol, 51 g (1.12 mol) formamide and 1200 ml anhydrous ether. The mixture was stirred vigorously at room temperature 20-26 ° C under a nitrogen atmosphere, then 157 g (1.12 mol) of benzoyl chloride in 50 ml of anhydrous ether was added dropwise via a dropping funnel. Addition takes 50 minutes.

The reaction mixture is stirred for an additional 60 minutes at room temperature, the ether is removed by pouring and 300 ml of acetic anhydride in 500 ml of anhydrous ether are added. The mixture was stirred at room temperature for 30 minutes. The precipitate was allowed to settle and the mixture of ether and acetic anhydride was decanted. The solid was collected by filtration, washed with 500 mL of ether, and dried in vacuo at 25 ° C with potassium hydroxide for 2 hours to give 130 g of benzylformimidate-40 hydrochloride as a white solid in 67% yield.

The product was analyzed by NMR: δ (DMSO) 5.7 (s, 2H, OCH), 7.5 (s, 5H, O), 9.0 (s, 1H, NC = N). The product is thermally unstable and decomposes to formamide and benzoyl chloride at 0 ° C and above. However, when stored 45 at -20 ° C for 2 months, no decomposition of the product could be detected.

Step B - Process for the preparation of thienamycin derivatives

Thienamycin (6 liters of an aqueous solution at pH 6.5, ie a fermentation broth containing 28 g of thienamycin) was placed in a 12 liter beaker and cooled to 0 ° C. The beaker is equipped with a pH meter and an efficient high speed stirrer. ₅₅ The pH was adjusted to 8.5 by careful addition of 3N KOH · which is added with vigorous stirring, added dropwise via syringe. 6 equivalents of approximately 100 g of solid benzylformimidate hydrochloride are then added in portions while maintaining the pH at 8.5 ± 0.3 by adding 200 ml of 3N potassium hydroxide by syringe. The addition takes 3 to 5 minutes. The reaction mixture is then stirred for 6 minutes at 0 ° C, and then liquid chromatography is carried out to confirm completion of the reaction. Thereafter, 1N hydrochloric acid is added to the solution until pH 7. The volume of the reaction mixture is measured and the solution is analyzed by ultraviolet radiation. The neutralized reaction mixture is concentrated to a concentration of 15 g / l by osmosis at a temperature below 10 ° C. The volume of the concentrate is again measured, the pH adjusted to 7.2 to 7.4, if necessary. The concentrate is then filtered through a medium pore sintered glass funnel to remove solids.

Step C - Chromatography on Dowax 50 Wx2 to 20 g Thienamycin 750 to 1000 ml is applied at 0 ° C to the top of a pre-cooled 18 liter Dowex 50 W x 2 column in a potassium cycle with a grain size of 0.074 up to 0.038 mm, the column is eluted at 0-5 ° C with distilled deionized water at a flow rate of 90 ml / min and at a pressure of 0 to 30 MPa.

Pre-fractions of 4 liters, 2 liters and 1 liter were collected, followed by 18 fractions of 450 ml and a final fraction of 2 liters. Each fraction was examined in ultraviolet radiation at a 1: 100 dilution, the NH 3 OH extinction was omitted, the total amount of NFT in each fraction was counted. The starting and final fractions are examined for purity by liquid chromatography and it is possible to pool the high product fractions. Then the pH of the fractions so mixed is determined with a pH meter and bromothymol blue, then adjusted to a pH of 7.2 to 7.4, if necessary. An amount of 3 to 4 liters of the thus-collected fractions is obtained, which is analyzed by ultraviolet radiation and the total content of the formamidine derivative is determined. This is generally 75% of the total yield of 15-16 g. The fractions are concentrated by osmosis at a temperature below 10 ° C, preferably up to a concentration of 33 g / liter, optionally on a rotary evaporator at a temperature below 28 ° C. A total of 500 ml of concentrate is obtained.

Step D - Crystallization of N-formimidoylthienamycin

If necessary, the concentrate of the previous step is adjusted to a pH of 7.3 and the N-formimidoylthienamycin content is determined by ultraviolet radiation. The yield is 85 to 90%. The concentrate was filtered through a medium pore sintered glass funnel into a large Erienmeyer flask. Five volumes, 2200 mL of 3A ethanol are added and the solution is stirred at room temperature for 10 minutes and then at 0 ° C for 12 to 24 hours.

The crystals were collected by suction filtration and washed with one volume, 250 mL of 80% 3A ethanol at 0 ° C, and then once more with 25 volumes, 100 mL of 3A ethanol at room temperature. The crystals were dried under vacuum for 12 to 24 hours to give a total of 10 to 12 g of N-formimidoylthienamycin in 40% yield.

Analysis was carried out on 50 g of N-formimidoylthienamycin prepared as described above. analysis:

calculated: C: 45.42, H: 6.03, N: 13.24, S: 10.10%, found: C: 45.82, H: 5.72, N: 13.10, S: 10 , 14%. Annealing residue: calculated 0.5, found 0.47%. [?] ²⁵ = 89.4 TG = 6.8%, ultraviolet spectrum Z _max 300 nm, E% 328.

The combination of a novel chemical inhibitor produced by the process of the invention and a compound of the thienamycin family may form a pharmaceutical composition comprising the two substances together with a pharmaceutically acceptable carrier. Said substances are usually used in an amount such that the ratio is poor

The value of the thienamycin-to-inhibitor value was in the range of 1: 3 to 30: 1, preferably 1: 1 to 5: 1.

Both components may also be administered separately. For example, a compound of the thienamycin family may be administered intramuscularly or intravenously at a dose of 1 to 100 mg / kg / day, preferably 1 to 20 or 1 to 5 mg / kg / day in divided form, i. j. three to four times a day. The inhibitor may be administered separately orally or intramuscularly, optionally intravenously, in an amount of 1 to 100 mg / kg / day, preferably 1 to 30 or 1 to 5 mg / kg / day. The amount of both ingredients administered over one day is in the ratio indicated above.

Most preferably, known doses are administered by administering two crystalline compounds, one of which is N-formimidoylthienamycin and the other is (+) - (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, simultaneously in sterile aqueous intravenous injection, in the form of the sodium salt, by administering 250 to 500 mg of thienamycin and its ratio to heptenoic acid being about 1: 1, i.e. 250 to 500 mg of this acid is also administered. This is a human dose of about 80 kg and is administered 1 to 4 times a day, i.e., 3.1 to 25 mg / kg / day of each of these agents is administered.

The individual components, whether administered separately or together, require the use of a pharmaceutically acceptable carrier for oral administration in the form of capsules or tablets, or solutions or suspensions. The individual components may also be administered separately or co-dissolved in an injectable solvent. Suitable formulations for oral administration may include diluents, granulating agents, preservatives, binders, flavoring agents, and coatings. The composition for oral administration may comprise gelatin, starch, magnesium stearate and alginic acid, in which case it is most preferably compressed into tablets.

As mentioned above, parenteral administration of a compound of the thienamycin family is most preferred with concomitant parenteral or oral administration of the inhibitor.

Determination of the effectiveness of the antibacterial mixture

As mentioned above, metabolic degradation has been demonstrated in various animal species, such as mouse, rat, dog, chimpanzee and monkey Rhcsus, for thienamycin, its natural analogs and its semisynthetic derivatives. The extent of metabolism can be demonstrated by the fact that only a small amount of these substances can be recovered from the urine and, moreover, a short half-life in blood plasma. The essence of this degradation is cleavage of the lactam ring by renal dipeptidase (E. C. 3.4.1.11), which was first described in Bergmann M. and Schleich H., Z. Physiol. Chem., 205, 65 (1932), Greenstein, J.P., Advances in Enzymology, Vol. VUL Wiley-Interscience, (1948), New York, and Campbell B.J., Lin, Y-C., Davis, R.V., and Ballew E., The Purification and Properties of Particulate Renal Dipeptidase, Biochem. Biophys. Acta., 118, 371 (1966).

The ability of the compounds of formula (I) to suppress the renal dipeptidase activity has been demonstrated in vitro.

This demonstrated the ability of the compounds to inhibit the hydrolysis of glycyldehydrophenylalanine (GDP) by a soluble dipeptidase preparation isolated from pig kidneys. The procedure was carried out as follows: To 1 ml of a solution containing 50 mmol of MOPS ”(3- / N-morpholino / propanesulfonic acid) containing a buffer solution of pH 7.1, 5 µg of lyophilized enzyme was added and the test substance to a final concentration of 0.1. 1 mmol. After 5 min incubation at 37 ° C, GDP was added to a concentration of 0.05 mmol. The mixture is incubated for 10 minutes at 37 ° C, then the hydrolysis of GDP is measured by changing the optical density at 275 nm over time. Enzyme inhibition is determined by comparison with a standard experiment that contains no inhibitor and is expressed as K i constant. This is the concentration of inhibitor at which 50% inhibition of the enzyme occurs.

GDP is used as a substrate instead of thienamycin because the maximum rate of hydrolysis of this product by kidney dipeptidase is greater, so this method requires less enzyme. Otherwise, GDP has the same renal dipeptidase activity as thienamycin and K; is identical for both substrates.

In addition to the in vitro experiment, an in vivo experiment was performed to determine the ability of the test substances to inhibit metabolism, which should be manifested by an increase in the amount of thienamycin in the urine of mice. The experiment was performed by administering the test substance intravenously or subcutaneously at a dose of 10 to 100 mg / kg simultaneously with 10 mg / kg of thienamycin. The amount of thienamycin in the urine over 4 hours is then compared to that in the control group not receiving the test substance.

The amount of thienamycin in the urine was in all cases measured using a roller or discs as described in U.S. Pat. This bioassay using Staphylococcus aureus ATCC 6538 has a range of 0.04 to 3.0 pg / ml.

The invention will be illustrated by the following examples.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

In vitro experiments ml of a solution containing 50 mmol of MOPS ™ buffer, pH 7.1, is mixed with 5 µg of the renal enzyme of pigs and the test substance to a concentration of 1 mmol. After 5 minutes incubation at 37 ° C, GDP is added to a final concentration of 0.05 mmol. The mixture is incubated for 10 minutes at 37 ° C. Hydrolysis of GDP is measured by changing the optical density over time at 275 nm. Enzyme inhibition is determined by comparison to a standard assay without inhibitor and expressed as a percentage. The K i constant is determined as the concentration of inhibitor required for 50% inhibition of the enzyme. These values were obtained from a large number of in vitro experiments at concentrations causing both less and greater inhibition than 50%. The results are shown in Table I.

Table I

COOH h I

Compound R ³ - C = C - NHCOR ²

Dipeptidase inhibitor R ³ R ² Inhibition at 10 ^-4 M ki (ΜΜ) 1 CH2CH3 ^CH 3 98 0.18 ^Xcll 3 2 * ch ₃ / \ / ^CH 3 98 0.39 XII ^ 2 * CH, / X / ^CH 3 ^X CII ₃ 100 0.12 2b * ch ₃ / X / ^ch 3 19.8 ^ Cll ₃ 3 ch ₃ 92 1.7 CH ₃ -CH 2 -CH 4 ch ₂ ch ₃ 87 3.2 ^ ch ₃ ch ₃ -ClIjCn-CIIjCtCII,? 5 CH, 81 4.4 / V-- ^CH 3 6 ch ₃ 83 4.6 ^CH 3 ^ -CTI ₃ 7 ch ₃ -CH9 -CH 91 6 8 CH ₃ x> 80 6.2 9 ch ₃ -ch ₂ --Zj> 83 6.6 10 ch ₃ z ^{/ X} x -CH ₂ -CH-CH ₂ CH ₃ ch ₃ 97 9 11 ch ₃ 82 10 12 - (CH ₂ ) ₄ CH ₂ FROM'·/' 0.03 Cl 13 - (CH ₂ ) ₅ hr (CH ₃ ) ₃ from \ / ^c ' 14 - (CH ₂ ) ₃ bT (CH ₃ ) ₃ / X / ^CH 3 1.11 ^ C1I ₃ 15 H -CCHjl-S-CH-C-COO · N h} / X / ^CH 3 ^X C1I ₃ 16 / X / ^CH 3 NH ^X C11 ₃ 17 H - (^ -S-Gly-IIb-C CCXy / X / ^CH 3 0.21 N H * ^ Cll ₃ 18 ch ₃ -CH ₂ C (CH ₃ ) ₃ 75 20 19 ch ₃ - (CH ₂ ) ₆ CH ₃ 72 26 20 ch ₃ - (CH ₂ ) ₂ CH ₃ 69 30 21 ch ₃ - ( ^CH 2> 3 ----- 68 30

Table I - continued

R ³

R ²

Inhibition at 10 ^-4 M CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃ CH ₃

HOO-CH2CH2 ch ₃ ch ₃ ch ₃ ch ₃ ch ₃

CH ₃ (CH ₂ ) 4 ch ₃ (ch ₃ ) ₂ ch ch ₃

(CH ₂ ) ₃ CH ₃

- (CH ₂ ) 4 CH (CH ₃ ) ₂

-ch ₂ ch ₂

-CH ₂ - (CH ₂ ) ₃ CH ₃ - (CH ₂ j ₅ CH ₃ -CH (CH ₂ CH ₃ ) CH ₂ CH ₂ CH ₂ CH ₃ -CH (CH ₂ CH ₂ CH ₃ ) ₂ -CH (CH ₃ ) 2

FROM\

-CH ₂ - (pH-CH ₂ CH ₂ OCH ₃ ch ₃

CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 Cl

CH 2 -CH (CH ₃ ) 1

100

ki

Ä

6.5

2.6

0.45

0.54

0.86

SK 278319 Β6

Example 2

In vivo experiments

In vivo experiments on mice were performed as follows: Female Charles River CD mice of 5 g of 20 g were administered a subcutaneous chemical inhibitor.

After 2 minutes, an intravenous dose of thienamycin is administered. In addition, a group of control mice receiving only thienamycin is formed. Urinary thienamycin levels, as a percentage of the administered substance, were measured biologically. The results are shown in Table H. Compound numbers are identical to those in Table I. Compound 7 is 2-isovaleramido-2-butenoic acid and compound 10 is (Z) -2-cyclopropylcarboxamido-2-butenoic acid.

The 2-butene was subjected to in vivo experiments in combination with thienamycin (THM) in mice, according to the method of Example 2. The results are shown in Tables m and IV.

TabuľkaIH

Effect of co-administration of 2-isovaleramido-2-butenoic acid (compound 7) on isolation of thienamycin from mouse urine ^{<a :)}

Table Π

compound Dose mg / kg of compound Dose mg / kg thienamycin % of thienamycin isolated from urine 7 50 10 53 7 10 10 53 10 50 10 56 inspection - 10 25-30

Method of submission ^ ’ Dose mg / kg Isolation of THM from urine zlúčeninina7 THM zlúčeninina7 THM in % - IV or SC - 10 30 ± 5 SC SC 0.3 10 33 SC IV 2 10 42 SC SC 2 10 47 SC IV 10 10 53 SC SC 50 10 54 SC IV 50 10 53 SC SC 80 10 59 SC SC 100 10 81

Example 3

2-Isovaleramido-2-butenoic acid (compound 7) and (Z) -2- (2,2-dimethylcyclopropanecarboxamido) - (a) female Charles River CDi mice weighing 20 g (b) co-administration

SC subcutaneous

IV intravenously

Table IV

Simultaneous administration of (Z) -2- (2,2dimetylcyklopropánkarboxamido) -buténovej (compound 2) for the isolation of thienamycin from the urine of mice ^'a)

process administration ^ Dose mg / kp Isolation of THM from urine in% zlúčenínina7 THM Compound? THM SC SC SC SC SC SC SC SC SC SC SC 0.1 0.3 1 10 30 10 10 10 10 10 10 30 ± 5 35 40 46 60 73

(a) Female Charles River CDi mice weighing 20 g (b) co-administered

SC subcutaneous

Example 4

In another experiment in mice, it was shown that the systemic antibacterial activity of thienamycin was increased approximately three-fold by the administration of 2-isovaleramido-2-butenoic acid, as shown in Table V.

Table V

Effect of concomitant administration of 2-isovaleramido-2-butenoic acid on the systemic effectiveness of thienamycin in Staphylococcus aureus infection

ED ₅₀ , mg / kg THM independently + 100 mg / kg inhibitor 0.2 0.06

Example 5

A male Beagle strain was used in an experiment to determine the effect of dipeptidase inhibitors on the recovery of N-formimidoylthienamycin in urine. In a control experiment, the dog was administered 5 mg / kg of N-formimidoylthienamycin intravenously without inhibitor. In the second experiment, the same amount of N-formimidoylthienamycin was used, but in addition three doses of (Z) -2-isovaleramido-2-butenoic acid, each at 20 mg / kg, were administered. The first dose was given just after injection of N-formimidoylthienamycin, the second dose after 40 minutes and the third dose after 60 minutes. In a third experiment, 2 mg / kg of (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-butenoic acid was administered once, just prior to the injection of N-formimidoylthienamycin. The results are shown in Table VI.

Table VI

Isolation of N-formimidoylthienamycin from urine 3 hours after intravenous administration of 5 mg / kg in a Beagle dog

Substance under study % recovery from urine N-formimidoyltienamycín 7.8 + (Z) -2-isovaleramido- 46 2-butenoic acid + (Z) -2- (2,2-dimethylcyclo- -propánkarboxamido) -2-butenoic acid 53

Section 2 - Examples demonstrating the chemical production of inhibitors

Compounds with inhibitory activity can be prepared by direct condensation of the corresponding 2-keto acid or ester and amide of formulas (III) and (IV) r ³ ch ² cc2r + r ² cnh ² (III) (IV) wherein

R ² and R ³ have the abovementioned meaning and

R is hydrogen or alkyl.

Compounds of formula (I) are thus obtained.

The reaction is usually carried out by mixing the acid with the amide in a ratio of 1 to 4: 1 in an inert solvent, for example toluene or methylisovalerate, and refluxing with azeotropic removal of water for 3 to 48 hours, preferably 5 to 4 hours. 24 hours. After cooling, the product is obtained in crystalline form from the solution, but the resulting product can also be isolated by extraction using a base. The product can be recrystallized in a known manner. In the condensation of ketoesters, a small amount of p-toluenesulfonic acid must be added as a catalyst. This catalyst also accelerates the condensation reaction using some keto acids.

Another possible way of producing novel compounds with inhibitory activity is by reacting the α-amino acid in the form of tert-butyl ester with the acid chloride

O ₂ II O h

R - CC1 + - _CH2 - C - COO - C _{(CH3) 3}

NH?

(V) ² (VI) where

R ² and R ³ are as defined above.

The reaction is carried out in the presence of a base such as triethylamine in a solvent such as methylene chloride. The resulting N-acylated product of formula VH is then oxidized by treatment with tert-butyl hypochlorite followed by addition of sodium methoxide. In this way a 2-methoxy derivative of formula (VH1) and / or an α, β-unsaturated ester of formula (EX) is obtained. By further treatment with anhydrous hydrochloric acid, compounds of formula (VIH) or (IX), or mixtures thereof, are converted to the desired α, β-unsaturated free acid of formula (H)

OCH

I 3

R ₂ CHCO ₂ CH C (CH _{3) 3} R ^j CCO ₂ CH ₂ C (CH _{3) 3}

NHCR ² NHCR

IIII oo (VIII) (VIII)

R ³ CH = CCO ₂ C (CH ₃ ) ₃

NHCR ² II o (K) where

R ² and R ³ are as defined above.

Some compounds in which R ³ carries a terminal substituent, namely an amino group, a quaternary nitrogen atom, a thio derivative, an alkoxy group, a guanidine group, an acyloxy group or a cyano group, can easily be prepared from an intermediate having a bromine atom in the terminal position. Compounds with substituted amino, such as formamidone, ureido, and acylamido (acetamido), can be prepared from compounds with amino by reaction with benzylformimide hydrochloride, potassium cyanate and an appropriate acid anhydride, for example acetic anhydride.

The invention will be explained by means of the following examples, illustrating the production of active substances.

Some compounds in which R ³ has a terminal substituent, namely an amino group, a quaternary nitrogen atom, a thiol group or a carboxyl group, can be obtained as a derivative from an intermediate in which the terminal substituent is a bromine atom. This intermediate can be represented by the general formula (X)

H COOH ¹

Wherein n is the number of carbon atoms in the desired hydrocarbon chain, ie, an integer of 3 to 7 and R ¹ and R ² are as defined above.

If they have obtained a compound in which R ³ is a group as a terminal trimethylammonium group, the intermediate is allowed to react with trimethylamine. If the amino group is to be obtained, this intermediate is reacted with ammonia. In the case of the desired guanidine group, the reaction is carried out with guanidine and the intermediate is reacted with cysteine hydrochloride or the corresponding mercaptan to produce the thio derivatives including the 2-amino-2-carboxyethylthio group. When an amino derivative such as formamidine, ureido or acylamido is to be obtained, compounds containing an amino group can be used and reacted with o-benzylformimidate hydrochloride, potassium cyanide hydrochloride and an appropriate acid anhydride such as acetic anhydride.

Another possible process for the preparation of compounds in which R ³ contains a thio group as the terminal group is a process in which the chloro keto ester of formula (XI) is used as an intermediate.

Cl- (CH ₂ ) _n -CE ₂ -C-CO ₂ R (X 1) wherein

R and n are as defined above, and this compound is reacted with an appropriate amide of formula (IV) wherein

R ² is defined as above, in toluene at reflux in the presence of a catalytic amount of p - toluenesulfonic acid. The resulting product is subjected to hydrolysis to form an acid and the chlorine atom is removed by treatment with the appropriate mercaptan. This reaction makes it possible to use a chiral amide of formula (IV), thereby allowing the preparation of a functional side chain. In addition, it is possible to convert the mixture of Z and E isomers after condensation directly to form Z by adding acid to pH 3 and heating to 90 ° C for 3 minutes. Isolation is quick and easy.

Example 6

(Z) -2-Isovaleramido-2-butenoic acid

A solution of 1.07 g (10.5 mmol) of 2-oxobutyric acid and 0.71 g (7.0 mmol) of isovaleramide in 15 ml of toluene is stirred at reflux while the water is collected in a Dean-Stark apparatus. After 5 hours, the solution was cooled to crystallize. After settling, the solid was collected by filtration, washed with toluene and then with methylene chloride. 0.47 white crystals are obtained, m.p. 172-174 ° C after pre-softening. The material was recrystallized from diisopropyl ketone. Thin layer chromatography using toluene / AcOH 4: 1 showed only traces of the second isomer. In a yield of 25% 0.32 g of white crystals with a melting point of 175 ° C after preliminary softening was obtained. The NMR spectrum showed exclusively the Z-isomer.

Analysis for C 8 H 11 NCL calculated: C: 58.36, H: 8.16, N: 7.56%, found: C: 58.59, H: 8.55, N: 7.43%.

Example 7

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -2-pentenoic acid

A solution of 2-oxovaleric acid (1.74 g, 15 mmol) and 2,2-dimethylcyclopropanecarboxamide (1.13 g, 10 mmol) in toluene (20 ml) was heated to reflux while stirring while retaining the water in a small Dean. -Stark your device. After 20 hours, the solution was cooled and a slow stream of nitrogen was passed. Once a portion of the solvent has evaporated, crystallization occurs. The solid obtained was collected by filtration and washed with toluene and a small amount of diethyl ether. In this way, 0.63 white crystals having a melting point of 154.5 to 155.5 ° C are obtained in a yield of 30% after preliminary softening. Thin layer chromatography with toluene / AcOH 4: 1 showed only a slight trace of the other isomer. The NMR spectrum showed only the Z configuration.

Analysis for C 11 H 17 NO ₃ calculated: C 62.53, H: 8.11, N: 6.63%, found: C: 62.86, H: 8.27, N: 6.75%.

Example 8

(Z) -2- (3-Cyclopentylpropionamido) -2-butenoic acid

A solution of 3-cyclopentylpropionamide (1.41 g, 10 mmol) and 2-oxobutyric acid (1.53 g, 15 mmol) was stirred and refluxed using a small Dean-Stark apparatus. After 8 hours, the solution was cooled to give a massive crystallization. The solid was collected by filtration and washed with toluene and methylene chloride. 1.44 g of white crystals are obtained, m.p. 180.5-182 ° C after pre-softening. This material was recrystallized from methyl ethyl ketone. In a yield of 28%, 0.63 g of white needle-like crystals with a melting point of 184-185 ° C are obtained after a preliminary softening. Thin layer chromatography using toluene / AcOH 4: 1 showed only one spot, the NMR spectrum showed only pure Z-isomer. Analysis for C 11 H 19 NO ₃ calculated: C: 63.97, H: 8.50, N: 6.22%, found: C: 63.99, H: 8.67, N: 6.27%.

Example 9

(Z) -2- (2-ethylhexanamido) -2-butenoic acid 2-ethylhexanoyl chloride was added dropwise with stirring with 25 ml of cold concentrated ammonium hydroxide to give an immediate precipitate. The mixture was stirred for 2 hours, then filtered and the precipitate was air dried to give 6.5 g of the amide. 1.4 g (10 mmol) of this amide and 1.5 g of oxobutyric acid (15 mmol) were heated to reflux in 25 ml of toluene for 15 hours while removing water. The reaction mixture was then cooled and partially evaporated with a stream of nitrogen. After standing for 3 hours, the resulting product crystallized. The crystals were collected, washed three times with toluene and air dried. 1.13 g of the title compound of melting point 160 DEG-162 DEG C. are obtained in a yield of 50%. The NMR spectrum was consistent with the proposed structure and showed less than 5% of the E-isomer. Thin layer chromatography using toluene / AcOH 4: 1 showed only one spot. Analysis for C ₂ I1 iNO _{2 3} Calculated: C: 63.40, H: 9.30, N: 6.16% Found C: 63.63, H: 9.43, N: 5.88%.

Example 10

(Z) -2- (2,2-Dimethylcyclopropanecarboxamido) 2-butenoic acid

2.53 g (15 mmol) of 2-oxobutyric acid, 1.13 g (10 mmol) of 2,2-dimethylcyclopropanecarboxamide and 20 ml of toluene were stirred and refluxed for 10 hours. After cooling, the crystalline solid was collected by filtration, washed with toluene (3 x 10 ml) and dried to give the title compound (1.06 g), mp 140-141 ° C. Thin layer chromatography using toluene / AcOH 4: 1 gave only one spot, the NMR spectrum being consistent with the proposed structure.

Recrystallization from EtOAc by drying afforded 0.533 g of the title product, m.p. 142-143.5 ° C, homogeneous by thin layer chromatography.

H, 7.67; N, 7.10%. Found: C, 60.92; H, 7.71; N, 7.38%.

Example 11 (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -2-hexenediic acid

A mixture of 1.0 g of 2,2-dimethylcyclopropanecarboxamide, 2.4 g of 2-oxoadipic acid and 25 ml of methylisovalerate is heated at reflux for 4 hours while the water is removed with a modified Dean-Stark apparatus containing a molecular sieve ( 4A). After standing overnight at room temperature, the crystalline precipitate was collected by filtration, washed with ether and recrystallized from ethyl acetate to give 0.23 g of the title product, m.p. 163-165 ° C. The NMR spectrum was consistent with the proposed structure.

H, 6.71; N, 5.49. Found: C, 56.20; H, 6.83; N, 5.32.

Example 12

(Z) -2- (2,2-Diethylcyclopropanecarboxamido) -2-butenoic acid

A mixture of 2.3 g of 2-oxobutyric acid, 2.0 g of 2,2-diethylcyclopropanecarboxamide and 25 ml of toluene was heated at reflux for 16 hours while removing water with a modified Dean-Stark apparatus containing a molecular sieve (4A). ). The product does not precipitate upon cooling. Ether (25 ml) was added and the mixture was extracted three times with saturated aqueous sodium bicarbonate. The extracts are decanted and acidified with concentrated hydrochloric acid. A gummy precipitate forms and crystallizes after trituration with water. Recrystallization gave the title compound (0.31 g), m.p. 129-130 ° C. The NMR spectrum was consistent with the proposed structure.

Analysis for C _{2 HI} NO _3: C: 63.98, H: 8.50, N: 6.22% Found C: 64.01, H: 8.62, N: 6.21%.

Example 13

2- (2,2-Dimethylcyclopropanecarboxamido) -2-hexenoic acid

Stage A:

DL-norleucine tert. butyl

The general procedure is described in R. Roeske, J. org. Chem. 28, 1251 (1963).

To a suspension of 9.82 g (75 mmol) of DL-norleucine in 80 ml of dioxane in a 500 ml ice-cooled bottle was slowly added 8 ml of concentrated sulfuric acid. The resulting mixture was cooled in a dry ice bath and 80 ml of quenched isobutylene (80 ml) was added. The mixture was then allowed to warm to room temperature and shaken at a self-exerting pressure for 23 hours. After most of the isobutylene was evaporated, the slightly cloudy solution was cooled in ice and then added to a cold mixture of 400 ml of 1 N sodium hydroxide and 500 ml of diethyl ether. After shaking in a separatory funnel, the layers are separated and the aqueous fraction is washed with a further 100 ml of diethyl ether. The diethyl ether solution is shaken with 150 ml of 0.5 N hydrochloric acid. The acidic aqueous fraction was treated by the addition of 2.5 N sodium hydroxide until a strongly basic reaction and then shaken with 250 ml of diethyl ether. The ether solution was dried over magnesium sulfate, filtered, and concentrated on a rotary evaporator. The mixture was then evaporated under high vacuum in a steam bath to give 9.04 g of a clear colorless oil in a yield of 65%. The NMR spectrum showed only traces of dioxane. Thin layer chromatography with chloroform: methanol (9: 1) showed a single spot.

Step B: N- (2,2-dimethylcyclopropanecarbonyl) -DL-norleucine-tert. butyl

To a solution of DL-norleucine-tert-butyl ester (8.98 g, 48 mmol) and triethylamine (5.05 g, 50 mmol) in methylene chloride (100 ml) was added dropwise a solution of 6.39 g over 75 minutes with stirring in an ice bath. (48 mmol) of 2,2-dimethylcyclopropanecarbonyl chloride (M. Elliot and N. MR. James, British Patent No. 1,260,847 (1972)) in 50 mL of methylene chloride while the water is collected through a drying tube. During the addition triethylamine hydrochloride precipitates, especially in the final phase. The ice was gradually melted and the mixture was allowed to warm to room temperature. After 16 hours, the mixture was shaken with 200 ml of 0.5 N hydrochloric acid. The methyl chloride fraction was washed with a further 200 ml of 0.5 N hydrochloric acid, then twice with 200 ml of 0.5 N sodium hydroxide and finally with 200 ml of water. Then, the methylene chloride fraction was dried over magnesium sulfate, charcoal was added and the fraction filtered through Celite. Then, the filtrate is evaporated on a rotary evaporator, ultimately under high vacuum. 11.93 g of a light orange oil are obtained in a yield of 88%. Thin layer chromatography using 2: 1 hexane / ethyl acetate showed a single spot. The NMR and IR spectrum were consistent with the proposed structure. After standing for several days, the unused portion of the material crystallizes and then has a melting point of 52-65 ° C.

Stage C:

T. butyl 2- (2,2-dimethylcyclopropanecarboxamide) -2-metoxyhexanoát

The basic procedure was described by H. Poisel and V. Schmidt, Chem. Ber., 108, 2547 (1975).

To a solution of 6.37 g (22.5 mmol) of N- (2,2-dimethylcyclopropanecarbonyl) -DL-norleucine-tert. of butyl ester in 35 ml of diethyl ether, while stirring at room temperature under a nitrogen atmosphere and in the dark, is added 2.69 ml (2.45 g, 22.5 mmol) of tert-butylhypochlorite. After 15 minutes a solution of sodium methoxide, prepared by dissolving 0.52 g (22.6 mmol) of sodium in 35 ml of methanol, was added. The mixture was stirred at room temperature under nitrogen and in the dark. After 16.5 hours, the precipitated sodium chloride is collected by filtration. The filtrate is diluted with diethyl ether and washed successively with 50 ml of 0.5 N hydrochloric acid, 50 ml of saturated sodium carbonate solution and twice with 50 ml of water. The ether phase is dried over magnesium sulphate and then filtered. The filtrate was evaporated on a rotary evaporator and 6.45 g of the obtained pale yellow oil was subjected to preparative high pressure liquid chromatography to give 273 mg of one and 496 mg of the other diastereomer tert. butyl 2- (2,2-dimethylcyclopropanecarboxamido) -2-methoxyhexanoate, m.p. 114-118 ° C and 124-125.5 ° C, as well as 1.97 g of the Z-isomer of tert. butyl 2- (2,2-dimethylcyclopropanecarboxamido) -2-hexanoate as a colorless oil.

Stage D:

2- (2,2-Dimethylcyclopropanecarboxamido) -2-hexanoic acid

A solution of 0.84 g (3.0 mmol) of tert. of butyl 2- (2,2-dimethylcyclopropanecarboxamido) -2-hexanoate in 10 ml of diethyl ether saturated with anhydrous hydrochloric acid is allowed to stand at room temperature in the presence of a drying tube. After 17 hours the solution was evaporated and the remaining gummy material was dissolved in 10 ml of saturated sodium bicarbonate solution. The solution was washed with 15 ml of 0.5 N hydrochloric acid and dried over magnesium sulfate, filtered and evaporated to a viscous, oily liquid which was crystallized from toluene. 0.32 g of white crystals having a melting point of 119-122 ° C are obtained in this way. Thin layer chromatography using toluene / AcOH 4: 1 showed a single spot. The NMR spectrum shows a substantially pure Z-isomer.

In the event that the tert. butyl 2- (2,2-di-55-methylcyclopropanecarboxamido) -2-methoxyhexanoate with anhydrous hydrochloric acid in diethyl ether under similar conditions to give the same product.

Example 14

(+) - (Z) -2- (2,2-dimethylcyclopropanecarbonylamino) -2-octenoic acid, sodium salt

7.0 g of (+) - 2,2-dimethylcyclopropanecarboxamide, 14.7 g of 2-oxooctanoic acid ethyl ester, 50 mg of p-toluenesulfonic acid and 100 ml of toluene are placed in a ⁴³ -necked three-necked flask with a Dean-Stark apparatus containing molecular weight. 250 ml sieve. The mixture was heated to reflux with vigorous stirring for 27 hours. The resulting light yellow solution was cooled and concentrated in vacuo to ^3U nominal water bath at 45 ° C in the presence of water to easily remove the toluene. The gummy residue obtained is suspended in 230 ml of 2 N sodium hydroxide solution and then stirred at 30 DEG C. for 3 hours. Then the temperature was raised to 35 ° C for 2.5 hours to give ³³ a clear solution. This solution was cooled, 85 mL of methylene chloride was added and the pH was adjusted to 8.5 by the addition of 4 N hydrochloric acid while stirring. The organic layer was separated and discarded. 336 ml of the aqueous layer was analyzed by liquid chromatography to show that it consisted of 87% of the Z-isomer containing 37 mg / ml. An additional 85 ml of methylene chloride was then added and the pH adjusted to 4.5 with stirring. The organic layer was separated and the aqueous layer was re-extracted with 50 ml methylene chloride, the pH adjusted vu ZnO ³ to 4.5. The organic extracts were decanted, dried over sodium sulfate, filtered and evaporated to a gum. This residue is dissolved in 150 ml of isopropyl 16

The pH was adjusted to 8.2 by addition of 2N sodium hydroxide. The resulting solution was evaporated to an oily residue which was mixed with isopropyl alcohol to give a crystalline solid from which most of the water had been removed. This material was crystallized for 1 hour from 120 ml of isopropanol under ice-cooling, then filtered and washed with 50 ml of cold isopropanol and then with a large amount of acetone. The product is then dried for 2 hours at 60 ° C and 13.3 Pa to give 10.74 g of crystalline material in 63.2% yield, which is subjected to liquid chromatography to show that the product consists of mp 241-243 ° C

The starting (+) - 2,2-dimethylcyclopropanecarboxamide can be readily obtained by resolution of the D, L-acid followed by reaction with oxalyl chloride and ammonia to give the amide isomer.

For example, the procedure is as follows: 23.1 g of D, L-2,2-dimethylcyclopropanecarboxylic acid is suspended in 33 ml of water and the pH is adjusted to 8.0 by addition of 50% sodium hydroxide solution in an amount of about 10 ml. A solution of 38.4 g of quinine in a mixture of 60 ml of methanol and 30 ml of water containing 8 ml of concentrated hydrochloric acid and an additional 30 ml of water is then added, resulting in a pH of 7.1, a quinine hydrochloride solution.

Said solutions are added to the base mixture simultaneously while stirring. A gummy crystalline material is formed which is heated until two clear layers are formed and then vigorously stirred under cooling to give a crystalline product which is allowed to stand at room temperature for 2 days. The product was then collected by filtration, washed twice with 10 ml of water and twice with 10 ml of 50% methanol, then air dried under suction. In this way, 44.8 g of a strong quinine salt monohydrate with a melting point of 113 DEG-116 DEG C., [.alpha.] _D -94.3 DEG C., c = 1.0, chloroform is obtained in a yield of 48.7%. This material was recrystallized from acetone to give 24.35 g of product, mp 127-130 ° C. This purified quinine salt is converted to the acid by treatment with an aqueous base solution and chloroform followed by treatment with an acid, yielding 3.9 g of the acid with [α] _D + 146.0 ° with a yield of 96%.

This acid is converted to the amide as follows: 30.5 g of the (+) isomer of the acid is added via a dropping funnel to 54 ml of oxalyl chloride containing 1 drop of diemethylformamide, cooled to 10 ° C over 5-10 minutes. The mixture was stirred at room temperature overnight. A clear solution is obtained which is diluted with 100 ml of methylene chloride. Excess oxalyl chloride was removed by evaporation and the mixture was washed twice more with methylene chloride.

The resulting solution was diluted with an equal amount of methylene chloride and added successively via a dropping funnel to 100 mL of anhydrous liquid ammonia mixed with 100 mL of methylene chloride. A cooling bath consisting of dry ice and acetone is used during the addition. After the addition was complete, the cooling bath was removed and the mixture was stirred at room temperature for 0.5 hours. The mixture was filtered to remove precipitated ammonium chloride and evaporated to dryness. Yield 26.6 g of crude product in 88% yield. This product was redissolved in excess hot ethyl acetate and filtered through a preheated glass funnel with sinter to separate traces of ammonium chloride. Excess ethyl acetate was distilled off at atmospheric pressure. After half-volume distillation, 130 mL of heptane was added and ethyl acetate was further removed by distillation until the boiling point began to rise to 80 ° C. At this point, most of the product crystallizes. The mixture was no longer heated and allowed to cool slowly to 30 ° C, and then cooled in an ice bath to 0-5 ° C for 0.5 hour. In this way, the product is obtained as silver-white crystalline flakes, which are washed three times with a 1: 1.5 mixture of ethyl acetate and hexane and then dried in air to constant weight. In this way, with a total yield of 77.1% and a yield

87.6% relative to the crude product yields 23.3 g of product, m.p. 135-138 ° C. The melting point varies depending on the heating rate. The rotation is determined by dissolving 0.0543 g of product in 10 ml of chloroform, [α] D + 100.9 °.

Example 15

(Z) -2- (2,2-Dichlorocyclopropanecarboxamido) -2-butenoic acid

Stage A:

2,2-dichlórcyklopropánkarboxamid

7.1 g of 2,2-dichlorocyclopropanecarbonyl chloride (U.S. Pat. No. 3,301,896 of January 31, 1967) is added dropwise to 75 ml of concentrated ammonium hydroxide with vigorous stirring. The temperature of the reaction mixture is kept below 10 ° C in an ice bath. The mixture was stirred in an ice bath for 30 minutes and then at room temperature for an hour. The aqueous ammonia solution was evaporated under reduced pressure at a bath temperature of 50 ° C. The solid residue is extracted three times with 30 ml of hot ethyl acetate. The extracts are boiled up to a volume of 40 ml, then 20 ml of hexane are added. After cooling with ice, the solid is collected by filtration, washed with ethyl acetate / hexane (1: 1) and dried to give 2.7 g of 2,2-dichlorocyclopropanecarboxamide, m.p. 144-146 ° C. The NMR spectrum was consistent with the proposed structure.

H, 3.27; N, 9.10; Cl, 46.04%. Found: C, 31.26; H, 3.31; N, 9.11. Cl: 45.79%.

An additional 1.3 g of amide is obtained from the mother liquor, m.p. 143-145 ° C.

Stage B:

(Z) -2- (2,2-Dichlorocyclopropanecarboxamido) -2-butenoic acid

A mixture of 2-oxo-butyric acid (1.53 g, 15 mmol), 2,2-dichlorocyclopropanecarboxamide (1.54 g, 10 mmol) and toluene (10 ml) was heated at reflux for 12 hours while the water was removed with a modified Dean-Stark a molecular sieve apparatus (4A). 0.7 g of 2-oxobutyric acid is then added and the reaction mixture is refluxed for a further 12 hours. The mixture was cooled, diluted with toluene (20 ml) and extracted with saturated sodium bicarbonate solution (3 x 10 ml). The extracts were decanted, washed with ether and acidified with concentrated hydrochloric acid to pH 3 under pH control. A gummy product precipitates and gradually becomes a solid. This material was collected by filtration, washed

Drying and recrystallization from nitromethane gave 423 mg of (Z) -2- (2,2-dichlorocyclopropanecarboxamido) -2-butenoic acid, m.p. 188-189.5 ° C. The NMR spectrum was consistent with the proposed structure. 5

H, 3.81; N, 5.88; Cl, 29.78. Found: C, 40.48; H, 3.80; N, 5.91. Cl: 29.53%.

Example 16

(Z) -2- (2,2-Dichlorocyclopropanecarboxamido) -2-octenoic acid

A mixture of 2-oxobutyric acid (1.19 g, 7.5 mmol), 2,2-dichlorocyclopropanecarboxamide (0.77 g, 5.0 mmol) and toluene (5 ml) was reacted as described in the previous example. 537 mg of crude product are obtained, which is purified by conversion to the methyl ester in a mixture of methanol and boron trifluoride, by preparative thin layer chromatography on silica gel G using hexane / ethyl acetate (4: 1) by saponification of pure 7-methyl ester with 0.3 M lithium hydroxide. and methanol to give 88 mg of (Z) -2- (2,2-dichlorocyclopropanecarboxamido) -2-octenoic acid as a partially crystalline gum.

NMR spectrum (DMSO-d 6): δ 9.68 (s, 1H, NH), 6.50 δ (t, 1H, = ^H ),

0.87 δ (t, 3H, CH ₃ ).

Example 17

(Z) -8-Bromo-2- (2,2-diinetylcyclopropanecarboxamido) -2-octenoic acid

To a suspension of 14.4 g (0.3 mmol) of a 50% dispersion of sodium hydride in 360 ml of toluene, cooled in an ice bath and under a nitrogen atmosphere, was added a solution of 146 g (0.6 mol) 1.6 over 45 minutes. -dibromohexane and

57.6 g (0.3 mol) of ethyl 1,3-dithian-2-carboxylate in 120 ml of dimethyl fomiamide. The cooling bath was removed and the mixture was stirred at room temperature for 2 hours. The reaction mixture is washed three times with 210 ml of water, dried over magnesium sulphate and evaporated under reduced pressure to give 179.5 g of a yellow oil which contains the desired anhydrous dithiane, 1,6-dibromohexane and mineral oil. This crude material was used in the next step without purification.

To a suspension of 426 g (2.4 mol) of N-bromosuccinimide in 800 ml of acetonitrile and 200 ml of water over 45 minutes and a solution of crude dithiane in 100 ml of acetonitrile. The temperature of the reaction mixture is kept below 25 ° C with an ice bath. The mixture was stirred at 20 ° C for 10 minutes, then the dark red reaction mixture was poured into 2 L of 1: 1 hexane: methylene chloride. The solution is shaken twice with 400 ml of saturated sodium bicarbonate solution and once with 500 ml of water. 400 ml of saturated sodium carbonate solution are then added in small portions with vigorous stirring to produce carbon dioxide. Once the mixture has stopped foaming, the funnel is shaken and the aqueous phase is separated. The organic layer was extracted with 400 mL of saturated sodium carbonate solution

500 ml of water, and dried over magnesium sulfate. The solvent was evaporated under reduced pressure to give 133.8 g of a crude bromo ketoester containing 1,6-dibromohexane and a mineral oil. This crude material was used in the next step without purification.

A mixture of 133.8 g of crude bromo ketoester, 133 ml of 50% hydrobromic acid and 267 ml of acetic acid was heated to an internal temperature of 90 ° C for 75 minutes. The dark colored solution was then evaporated under reduced pressure to remove most of the acetic acid. The residue is dissolved in 500 ml of ether, washed twice with 100 ml of water and extracted three times with 200 ml of saturated sodium bicarbonate solution. The bicarbonate extracts are decanted, extracted twice with 100 ml of ether and acidified with concentrated hydrochloric acid. The resulting oil was extracted with ether (3 x 200 mL). The ether extracts are washed once with 100 ml of water and once with 100 ml of saturated sodium chloride solution and dried over magnesium sulfate. The ether was removed under reduced pressure to give 46.2 g of pure bromo keto acid. The product is homogeneous in silica gel thin layer chromatography using toluene / acetic acid 4: 1. The NMR spectrum was consistent with the proposed structure.

A mixture of 46.1 g (0.194 mol) of bromoketoacid, 17.6 (0.156 mol) of 2,2-dimethylcyclopropanecarboxamide and 450 ml of toluene was heated to reflux for 13 hours while the water was removed with a small Dean-Stark apparatus. After cooling, the clear reaction mixture was extracted four times with 100 ml of saturated sodium bicarbonate solution. The extracts are decanted, washed twice with 100 ml of ether and then adjusted to pH 3.5 by addition of concentrated hydrochloric acid, which is monitored by a pH meter. An oil precipitates which crystallizes rapidly. The solid was collected by filtration, washed well with water and dried. After recrystallization from acetonitrile, 22.5 g of (Z) -8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, m.p. 151-153 ° C, are obtained. The product is homogeneous in thin layer chromatography using acetic acid and toluene 4: 1. The NMR spectrum was consistent with the proposed structure. H, 6.67; N, 4.22; Br, 24.05%. Found: C, 50.66; H, 6.96; N, 4.45. Br: 23.95%.

In the same way, the following .beta.-bromofluorocarbons were prepared:

(Z) -6-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -hexenoic acid, (Z) -7-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -heptenoic acid, (Z) -9-bromo -2- (2,2-dimethylcyclopropanecarboxamido) -2-nonenoic acid (Z) -110-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -decenoic acid (Z) -8-bromo-2- (2 , 2-dimethylcyclopropanecarboxamide) octene.

Example 18

(Z) -8-Dimethylamino-2- (2,2-dimethyl-cyclopropanecarboxamido) -2-octenoic acid

A solution of 664 mg (2 mmol) of (Z) -8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid in 10 ml of a 40% aqueous dimethylamine solution was allowed to stand at room temperature for 4 hours. 3.5 x 20 cm Dowex 50Wx8 with a particle size of 0.147-0.074 mm in β-form and the column is eluted with water until the eluate has an acidic reaction, 200 ml of water is consumed, then the column is eluted with 300 ml of 2N solution The eluate was evaporated under reduced pressure to give 600 mg of a colorless glass, which was dissolved in 3 mL of ethanol, filtered and added dropwise to 200 mL of acetone with vigorous stirring. This material was collected by filtration, washed with acetone and dried to give 445 mg of (Z) -8-dimethylamino-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid as colorless, hygroscopic crystals. mp 101-112 [deg.] C. The product is homogeneous by thin layer chromatography on silica gel in a 4: 1: 1 mixture of butanol, acetic acid and water. The NMR spectrum was consistent with the proposed structure. H, 9.62; N, 8.91. Found: C, 61.03; H, 9.28; N, 8.67%.

The following 8-amino derivatives were prepared in a similar manner, "DCC" means 2- (2,2-dimethylcyclopropanecarboxamido): (Z) -10-dimethylamino-DCC-2-decenoic acid, (Z) -8-amino-DCC-2 -octenoic acid (Z) -8-dimethylamino-DCC-2-octenoic acid (Z) -7-dimethylamino-DCC-2-heptenoic acid (7th) -DCC-7- [N-methylpiperazinyl) -2 -heptenoic acid (Z) -DCC-8-pyrrolidino-2-octanoic acid (Z) -DCC-8- (N-methylpiperazinyl) -2-octanoic acid (Z) -8-allylamino-DCC-2- octenoic, (Z) -DCC-8-piperidino-2-octenoic acid, (Z) -DCC-8-propargylamino-2-octanoic acid, (Z) -8-N- [1-deoxy- (1-methylamino) (D) -D-glucityl] -DCC-2-octenoic acid (Z) -8- (1-adamantylamino) -DCC-2-octenoic acid (Z) -8-DCC-8- (2-hydroxyethylmethylamino) -2 -octenoic, (Z) -8-dialylamino-DCC-2-octenoic acid, (Z) -8 - [(carboxylmethyl) methylamino] -2- (2,2-DCC) -2-octenoic acid, (Z) -2- (2,2-DCC) -8-diethylamino-2-octenoic acid, (Z) -2- (2,2-DCC) -8- [tris- (hydroxymethy) (1) -methylamino] -2-octenoic acid, (Z) -2- (2,2-DCC) -1- (N-methylpiperazinyl) -2-decenoic acid, (Z) -2- (2,2- DCC) -8- [l- (phosphono) ethylamino] -2-octenoic.

Example 18A

(Z) -8 - [(Carboxymethyl) methylamino] -2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid

3.32 g of (Z) -8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, 1.0 g of CH 3 NH Clh-CO 2, 3.5 g of Na ₂ CO 3 and 30 ml of water are heated to 80 ° C under a nitrogen atmosphere for 1.5 hours. After purification, 1.0 g of the title compound is obtained. H, 8.57; N, 7.44%. Found: C, 54.40; H, 8.34; N, 7.16%.

Example 18B

(Z) -2- (2,2-Dimethylcyclopropanecarboxamido) -8- [1- (phosphono) ethylamino] -2-octenoic acid

Following the procedure of Example 18A, 335.1 mg of the brominated intermediate was reacted with 138.2 mg of 1-aminoethanephosphoric acid, 435 mg of sodium carbonate and 5 ml of water to give the final product with K = 0.16.

Example 19

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -8-methylthio-2-octenoic acid

A stream of methyl sulfide gas was bubbled through a solution of 162 mg (3 mmol) of sodium methoxide in 5 mL of methanol for 10 minutes while cooling in an ice bath. The solution was allowed to warm to room temperature and 332 mg (1 mmol) of (Z) -8-bromo-2- (2,2-dimethyl-cyclopropanecarboxamido) -2-octenoic acid was added.

The solution was heated to reflux for 30 minutes under a nitrogen atmosphere. Most of the methanol was evaporated under reduced pressure, the residue was dissolved in 10 ml of water and acidified with 2.5 N hydrochloric acid. The resulting oil was extracted three times with ether. The ether extracts were washed with water and saturated sodium chloride solution and then dried over magnesium sulfate. The ether was removed under reduced pressure to give a colorless oil which crystallized on standing. The product was recrystallized from ether / hexane to give (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-methylthio-2-octanoic acid (178 mg), m.p. 82-84 ° C. The product is homogeneous in thin layer chromatography using toluene / acetic acid 4: 1. The NMR spectrum was consistent with the proposed structure. Analysis for C 15 H 25 NO 3 S Calculated: C: 60.18, H: 8.42, N: 4.68, S: 10.69%, found: C: 60.36, H: 8.68, N: 4.59, S: 10.87%.

In a similar manner, the following compounds were prepared in which DCC is 2- (2,2-dimethylcyclopropanecarboxamido): (Z) -DCC-8-ethoxythiocarbonylthio-2-octenoic acid, (Z) -DCC-8- (1-methyl) -5-Tetrazolylthio) -2-octanoic acid, (Z) -DCC-7 - [[((methoxycarbonyl) methylAio] -2-heptenoic acid, (Z) -8-acetylthio-DCC-2-octenoic acid, (Z) -7 - [(2-amino-2-oxoethyl) thio] -DCC-2-heptenoic acid, 6- (L-2-amino-2-carboxyethylthio) -2- (2,2-DCC) -2-hexenoic acid (Z) -8- (carbomethoxymethylthio) -2- (2,2-DCC) -2-octenoic acid, (Z) -6- (carbomethoxymethylthio) -2- (2,2-DCC) -2-hexenoic acid (Z) -2- (2,2-DCC) -6- (phosphonomethylthio) -2-hexenoic acid.

Similarly, 7- (L-amino-2-carboxyethylthio) -2- (2,2-dimethylcyclopropanecarboxamido) -2-heptenoic acid can be obtained except that 185 mg (1.05 mol) of acid (Z) is dissolved. 7-Bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-heptene prepared as described in Example 17 in 2.02 ml of 2.0 N sodium hydroxide, and the mixture is degassed by bubbling for one minute. nitrogen gas. Then 185 mg (1.05 mol) of cysteine hydrochloride are added in one portion and the reaction mixture is stirred at room temperature under a nitrogen atmosphere for 3 hours. The reaction mixture is then applied to a 2 x 20 cm column containing Dowex 50 x 4 resin (0.147 to

SK 278319 Β6

0.05 ml (ET) and elute the column with 300 ml of water, then wash with 200 ml of 2N ammonia. The ammonia was evaporated under reduced pressure to give 284 mg of a yellow glassy product. This product was dissolved in 4 ml of ethanol and the insoluble material was filtered off. The filtrate is added dropwise to 150 ml of diethyl ether with vigorous stirring. The resulting solid was collected by filtration, washed with ether and dried to give 171 mg of the ninhydrin positive product and gave only one spot on TLC with Rf 0.6 using 4: butanol: ethanol: water. 1: 1st The nuclear magnetic spectrum agrees with the assumed structure.

H, 7.31; N, 7.81; S, 8.94%. Found: C, 52.55; H, 7.40; N, 7.89. S: 9.63%.

Example 19A

(Z) -7- (L-Amino-2-carboxyethylthio) -2- (2,2-dimethylcyclopropanecarboxamido) -2-heptanoic acid as the sodium salt

A. Grignard reaction to produce ethyl 7-chloro-2-oxoheptanoate mole of 1-bromo-5-chloropentane and 8 mL of magnesium was reacted in 960 mL of tetrahydrofuran at 25 ° C. The two reactants were introduced into a flask and bromochloropentane was added over 1 hour, then the mixture was allowed to stand for 2 hours. After completion of the reaction, the reaction solution, after cooling to -15 ° C, was added to 16 moles of diethyl oxalate in 1856 ml of tetrahydrofuran, bringing the temperature to 10 ° C. Subsequently, 3N hydrochloric acid is added and the temperature is maintained at 25 ° C. Evaporation gave ethyl 1-chloro-6-oxoheptanoate in 48.8% yield.

B. Condensation and hydrolysis

1017 g of (S) -2,2-dimethylcyclopropylcarboxamide,

2143.6 g of ethyl 7-chloro-2-oxoheptanoate, 9 liters of toluene and 12 g of p-toluenesulfonic acid are charged to a 22 liter flask and the mixture is heated to reflux while stirring. After 23 hours, liquid chromatography shows the expected product yield and 4 liters of toluene are evaporated under reduced pressure. Water was added, the mixture was neutralized to pH 7 by addition of 2N sodium hydroxide, and the mixture was distilled under vacuum to a volume of 5 liters.

Then the mixture is hydrolyzed by adding 1760 g of aqueous sodium hydroxide solution, using 4 liters of water and the hydroxide concentration is 50%, then the mixture is stirred overnight. To the flask was added 4 liters of methylene chloride and the pH was adjusted to 8.8 with hydrochloric acid. The unreacted amide crystallizes. The organic layer was separated from water and evaporated. The gummy residue is dissolved in 8 liters of water containing 720 g of sodium hydroxide, which is a 50% solution, and 1818 g of cysteine hydrochloride with 1 molecule of water, 2 kg of ice, 2484 g of 50% sodium hydroxide and liter of water.

After standing overnight at room temperature, the pH was adjusted to 3.0 by addition of concentrated hydrochloric acid and the resulting gummy suspension was heated to 95 ° C to give a clear solution. After 30 minutes, no isomer E could be detected by thin layer chromatography. After work-up and purification, 2060 g of product were obtained in a yield of 87%. This material was recrystallized from acetonitrile. 1500 g of recrystallized material was dissolved in 6 liters of water, then after addition of 910 ml of 3.88 N sodium hydroxide by neutralization to pH 7 and lyophilization in 5 yields 98.6%, the title compound was obtained with a total amount of 1569 g.

Analysis: Calculated: C: 50.52, H: 6.62, N: 7.36, S: 8.43, Na: 6.04%, Found: C: 50.71, H: 6.78, N : 7.49 S: 8.52, Na: 5.92%.

Example 19B

In a similar manner to Example 19, ('Z) -8 [(2-amino-2-oxoethyl) thio] -2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid can also be obtained using 3.3 g of the brominated intermediate. and 1.3 g of H 2 NC / = O / CH 2 OH in 50 mL of methanol. 1.6 g of product with a melting point of 127-128 ° C are obtained.

Example 20

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -8-trimethylammonium hydroxide-2-octenoic acid inner salt

A solution of 996 mg (3 mmol) of (Z) -8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid in 15 ml of a 25% aqueous trimethylamine solution was allowed to stand at room temperature for 3 hours. The reaction mixture is then poured onto a 2 x 25 cm column packed with IRA-410 having a particle size of 50-100 mesh in OH-form and eluted with water until the eluate has a basic reaction. The eluate was evaporated under reduced pressure to give 800 mg of a colorless glass. This material was dissolved in 20 mL of ethanol, filtered and diluted with 600 mL of acetone. The mixture was allowed to stand at room temperature overnight and then the crystalline solid formed was collected by filtration, washed with acetone and dried to give 720 mg of (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-trimethyl inner salt. ^42- ammonium hydroxide-2-octene in the form of hygroscopic crystals, m.p. 220-222 ° C. The product is homogeneous in silica gel thin layer chromatography using 4: 1: 1 butanol / acetic acid / water. The NMR spectrum ^{3 was} consistent with the proposed structure.

Analysis preCnH3oN ₂ 03: C: 65.77, H: 9.74, N: 9.02%; found: C 65.78, H: 9.98, N: 8.92%.

Similarly, the following quaternary derivatives can be obtained:

(Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-trimethylammonium hydroxide-2-octanoic acid inner salt, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-pyridinium hydroxide-2-octenoic acid, inner salt (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8- (2-hydroxyethyldimethylammonium hydroxide) -2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -10-trimethylammonium hydroxide-2- (2-hydroxyethyldimethylammonium hydroxide) inner salt octenoic acid, (Z) -8- (benzyldimethylammonium hydroxide) -2- (2,2-dimethylcyclopropanecarboxamido) -265-octanoic acid, inner salt of (Z) -10- (benzyldimethylammonium hydroxide) -2- (2,2-dimethylcyclopropanecarboxamide) ) -220

SK 278319 Β6

-decenoic, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -9-trimethylaminonium hydroxide-2-nonenoic acid, inner salt of (Z) -8- (2-dimethylaminoethyldimethylammonium hydroxide) -2- (2,2-) (Z) -2- (2,2-dichlorocyclopropanecarboxamido) -8-trimethylammonium hydroxide-2-octenoic acid, inner salt of (Z) -2- (2,2-dichlorocyclopropanecarboxamido) -dimethylcyclopropanecarboxamido) -2-octanoic acid.

Example 21

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -8-formamidino-2-octanoic acid

350 mg of (Z) -8-amino-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid are dissolved in 10 ml of water and the pH is adjusted to 8.5 by addition of 2.5 N sodium hydroxide. 947 mg of benzylformimidate hydrochloride are then added in small portions at room temperature over a period of 20 minutes, maintaining the pH in the range of 8-9 by the addition of 2.5 N sodium hydroxide. The mixture was stirred at room temperature for 30 minutes, then the cloudy reaction mixture was extracted three times with ether and applied to a 2 x 2.5 cm column packed with G50W-X4 having a particle size of 0.074-0.038 mm in Na ^- form. The column was eluted with water and the fractions containing product were combined and evaporated under reduced pressure. The material obtained is dissolved in water and applied to a 2 x 25 cm column packed with G1X8 in HCO 3 -form with a particle size of 0.074-0.028 mm. After washing with water, the fractions containing the pure product are decanted and evaporated under reduced pressure. The residue is dissolved in a few ml of warm ethanol, filtered and added dropwise to 200 ml of ether with vigorous stirring. After filtration and washing with ether, 243 mg of (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-formamidino-2-octenoic acid are obtained as an amorphous solid. The product is homogeneous by thin layer chromatography using a 4: 1: 1 mixture of n-butanol, acetic acid and water. The NMR spectrum is consistent with the proposed structure.

H, 8.59; N, 13.92. Found: C, 60.04; H, 8.64; N, 13.57%. C15H25N3O3.I / 3H2O requires C, 59.69; H, 8.59;

In a similar manner, the following amidine derivatives can be obtained:

(Z) -8-Acetamidino-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, (Z) -8-Benzylamidino-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -10-formamidino-2-decenoic, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8- (2-imidazoyl-2-ylamino) -2-octenoic acid.

Example 22

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -8-guanidino-2-octenoic acid

To a solution of 2 mmol of guanidine prepared from 432 mg of guanidine sulphate and 630 mg of barium hydroxide as octahydrate in 7 ml of water is added 332 mg (1 mmol) of 8-bromo-2- (2,2-dimethylcyclopropanecarboxainide) octoenoic acid and heated to 70 ° C. The reaction mixture was then applied to a 2 x 25 cm column packed with Dowex 50W-X8 in H ⁴ -form with a particle size of 0.147 to

0,074 mm. After elution with water, the product-containing fractions were combined and evaporated under reduced pressure. The residue was dissolved in a few ml of warm ethanol and added dropwise to 100 ml of ether with vigorous stirring. After filtration and washing with ether, 107 mg of (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-guanidino-2-octenoic acid is obtained as an amorphous electrostatic powder. The product is homogeneous in thin layer chromatography using a 4: 1: 1 mixture of n-butanol, acetic acid and water.

Nuclear Magnetic Resonance Spectrum (D ₂ O, NaOD): 6.48 δ (t, 1H, = ^H ), 3.10 δ (m, 2H, CHN -), 2.10 δ (m, 2H, ^--CH ₂ ) , δ

In a similar manner, the following guanidino compound can be obtained:

(Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8- (N, N-dimethylguanidino) -2-octenoic acid.

Example 23

(Z) -2- (2,2-Dimethyl-cyclopropanecarboxamido) -8-methyl-2-octenoic acid

To a solution of 2 m 43 mmol of sodium methoxide in 5 mL of methanol was added 332 mg (1 mmol) of 8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid. The solution was heated to reflux under nitrogen for 1 hour. The reaction mixture was evaporated under reduced pressure, the residue was dissolved in water and acidified with 2.5 N hydrochloric acid. An oil precipitated and was extracted three times with ether. The ether extracts were washed with water and saturated sodium chloride solution and dried over magnesium sulfate. Removal of the ether under reduced pressure gave a colorless oil which crystallized on standing. The crystals were recrystallized from ether / hexane to give 140 mg of (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-methoxy-2-octenoic acid, mp 71-72 ° C. The product is homogeneous by thin layer chromatography using toluene / acetic acid 4: 1. The NMR spectrum is consistent with the proposed structure.

Analysis for C 14 H 25 NO 4 calculated: C: 63.58, H: 8.89, N: 4.94%, found: C: 63.54, H: 9.12, N: 5.16%.

In a similar manner, the following compounds can be obtained:

(Z) -8-Cyano-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid, (Z) -7-Cyano-2- (2,2-dimethylcyclopropanecarboxamido) -2-heptenoic acid, (Z) -9-cyano-2- (2,2-dimethylcyclopropanecarboxamido) -nonenoic, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -7-sulfo-2-heptenoic acid, sodium salt (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-sulfo-2-octenoic acid, (Z) -2- (2,2-dimethylcyclopropanecarboxamido) -8-hydroxy-2-octenoic acid, (Z) -8-acetoxy 2- (2,2-dimethylcyclopropanecarboxamide) -2-octenoic.

(Z) -8-Cyano-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid can be obtained by using 332 mg of 8-bromo-2- (2,2-dimethylcyclopropanecarboxamido) -2-octenoic acid and 100 mg of sodium cyanide in 2 ml of dimethyl sulfoxide and the mixture is heated to 80 ° C for 30 minutes. After extraction and purification, 102 mg of a colorless solid are obtained, m.p. 99-103 ° C.

H, 7.97; N, 10.06%. Found: C, 64.69; H, 8.14; N, 9.41%.

By the method of the preceding examples, it is also possible to obtain all the compounds which are listed in the following Tables I, H and ΙΠ together with their physicochemical constants.

Table I of (Z) -2-acylamino-2-butenoic acid

H CO H

compound R 2 mp ° C crystallization. solvent ^b yield% formula analysis ⁰ K., μΜ or% inhibition at 100 μΜ 1 2 CH, CH 2 CH 3 151-155 ° ¹ 149-150.5 I 26 C7H11NO3 C, H, N (12%) (43%) 3 -CH2CH2 239-241 Π 40 C 14 H ₂ O ₂ 06 C, H, N 10 4 (CH ₂ ) ₂ CH, with decomposition 141-142 Π 23 CoHiaNO ;, C, H, N 30 5 (CH 2) ₃ CH ₃ 153-154 IV 18 CíjHisNOd C, H, N 32 6 (CH ₂ ) ₄ CH ₃ 141-142 IV 23 C ₁₀ H 17 NO ₃ C, H, N (54%) 7 (CH ₂ ) ₅ CH ₃ 142 I 8 ChH ₁₉ NO ₃ C, H, N (49%) 8 (CH ₂ ) ₆ CH ₃ 141-142 IV 23 C12H21NO3 C, N, H ⁶ 26 9 (ch ₂ ) ₇ ch ₃ 145-147 IV 30 C 13 H ₂₃ NO ₃ C, H, N 32 10 (CH ₂ ) s CH ₃ 145-146 in 11 CÄjNOj C, H, N (58%) 11 (CH ₂ ) ₉ CH ₃ 141-142 IV 19 C ₅ H ₂₇ NO ₃ C, H, N 19 12 (CH ₂ ) k> CH ₃ 141-142 IV 24 C16H29NO3 C, H, N 35 13 (CH ₂ ) ₁₂ CH ₃ 149-151 IV 34 C ₁₈ H ₃₃ NO ₃ C, H, N (45%) 14 (CH ₂ ) ₁₆ CH ₃ 140-141 IV 25 C ₂₂ H ₄₁ NO ₃ C, N, H ^f (12%) 15 CH _{(CH3) 2} 184 to 184.5 IV 11 CgHisNOä C, H, N 31 16 CH ₂ CH (CH ₃ ) ₂ 176 VI 27 CsHiíNOs C, H, N 6 17 C _{(CH3) 3} 182 to 183.5 rv 14 C ₉ H ₁₅ NO ₃ C, H, N (0%) 18 (CH ₂ ) ₂ CH (CH ₃ ) ₂ 164-165 rv 13 C10H17NO3 C, H, N 14 19 CH ₂ CH (CH ₃ ) CH ₂ CH ₃ 168.5 to 169.5 rv 27 c ₁₀ h ₁₇ no ₃ C, H, N 10 20 CH ₂ C (CH ₃ ) ₃ 191-192 rv 47 C10H17NO3 C, H, N 20 21 CH _(CH3) CH _{(CH3) 2} 197-198 in 34 C10H17NO3 C, H, N 3.3 22 CH (CH ₂ CH ₂ CH ₃ ) 2 185-186 rv 53 C12H21NO3 C, H, N (13%) 23 CH (CH ₂ CH ₃ ) (CH ₂ ) ₃ CH ₃ 158-160 rv 43 C12H21NO3 C, H, N (33%) 24 (CH ₂ ) ₄ CH (CH ₃ ) ₂ 146.5 to 147 in n 7 C12H21NO3 C, H, N (57%) 25 CH ₂ CH (CH ₃ ) CH ₂ C (CH ₃ ) ₃ 136-138 IV 21 C13H23NO3 C, N, H ⁸ 44 26 CH 2 CH (CH ₃ ) (CH ₂ ) ₃ CH (CH ₃ ) 2 144-145 IV 55 C ₁₄ H ₂₅ NO ₃ C, H, N 4 27 (CH ₂ ) ₂ CH = CH ₂ 143-145 rv 17 C7H13NO3 C, H, N (57%)

Table I - continued

compound ri mp ° C rats. solvent? yield% formula analysis ⁰ K i, μΜ or% inhibition at 100 μΜ 28 CH = (CH3) ₂ 183-184 in M 30 C9H13NO3 C, H, N 28 29 ch = chch = chch ₂ 198-199 wine, v 11 CkjHijNOj H, N; C ^h (0%) 30 (CH ₂ ) ₈ CH = CH ₂ 135-137 IV 27 C ₁₅ H ₂₅ NO ₃ C, H, N 34 31 COH, ¹ 70 32 4-CH ₃ C ₃ Hi 199-200 IX CkIIbNOj C, H, N (0%) 33 LCQ 212-215 x 98 CisHijNOs C, H, N (3%) 34 about 140 to 141.5 XI 9 C ₉ H ₉ NO ₄ C, H, N (57%) 35 CH ₂ C 6 H ₅ 195-197 ΧΠ 6 C ₁₂ H ₁₃ NO ₃ C, H, N (21%) 36 (CH _{2) 2} C 6 H ₅ 188-189 in 18 C13H15NO ₃ C, H, N (30%) 37 (CH _{2) 3} C 6 H ₅ 130-132 IV 49 C ₁₄ H ₁₇ NO ₃ C, H, N 11 38 132-135 IV 55 C ₁₂ H ₁₃ NO ₃ S C, H, N 10 39 CH = CH ₅ 198.5-201 IV 27 ^k c ₁₃ h ₁₃ no ₃ C, H, N (0%) 40 ch ₂ ci 166-168 ¹ (52%) 41 CHiOCíHj 127.5-128 IV 37 Ci ₂ Hi ₃ NO ₄ C, H, N (3%) 290 za 42 ch ₂ nh ₂ decomposition (6%) 43 CH ₂ NHCO (CH _{2) 3} JCH 164 ΧΠ 18 C ₁₀ H 16 N ₂ 0 ₄ C, H, N (5%) 44 CH ₂ NHCOCH ₂ (CH ₃ ) ₂ 144 xn 25 CuHisN ₂ O ₄ C, H, N (5%) 45 ch ₂ sch ₂ ch ₃ 114 to 115.5 IV 7 CsHijNO ^ C, H, N 31 46 133.5-134 XI 19 C ₁₁ H ₇ NO ₄ C, H, N (51 “/ ·) 47 (CH ₂ ) ₃ CO ₂ CH ₂ CH ₃ 10-103 in n 9 CnHnNOí C, H, N 39 48 (CH ₂ ) ₃ CON (CH ₃ ) ₂ 137.5 to 139 in M 26 CnH _l8 N ₂ O ₄ C, H, N (46%) 49 (CH ₂ ) i NHCOCH ₃ 155-157 xn 20 C ₁₀ H ₁₆ N ₂ 0 ₄ .1 / C, H, N 102 / 4H ₂ O 50 (CHjjsNHCOCHjC ^ CH ^ 132-135 xn 18 C ₂₂ H ₂₂ N ₂ O ₄ C, H, N 52 51 (CH ₂ ) ₄ CN 102-109 ^r XI 5 CkHi ^ Oj. .0.125 H ₂ O ₃ C, H, N 25 52 (CH ₂ ) ₄ CO ₂ CH ₂ CH ₃ 123-124 in n 18 C12H19NO5 C, H, N 18 53 (CH ₂₎ SBR 132-135 IV 8 C ₁₀ H 13 BrNO ₃ C, H, N 19 54 (CH ₂ ) 5 NHCOCH ₂ CH ₃ 108-110 xn 12 C 13 H ₂₂ N ₂ O ₄ . C, H, N 41 ., 1 / 4H ₂ O 55 CH ₂ CH (CH ₃ ) (CH ₂ ) ₂ OCH ₃ 105-107 rv 21 c _n H19NO ₄ C, H, N 9 56 CH ₂ CH (CH ₃ ) (CH ₂ ) ₃ C (OH) (CH ₃ ) ₂ 115-117 IV 20 C ₁₄ H ₂₃ NO ₄ C, N; H ⁸ 6.7 57 (CH ₂ ) ₈ CO ₃ H 155-158 xn 2 C ₁₄ H ₂₃ NO ₅ C, H, N 28 58 4-OCH ₃ CeH 4 201-203 x 60 C ₁₂ Hi3NO ₄ H, N; C ' (0%) 59 2-ClCeH, 207-209 xn 22 CnHioClNOs C, H, N (25%) Cl 60 CIC-3, | H <> 188-190 xn 34 CnHioCINOj C, H, N (22%) Cl 61 4-C1CJL, 227 with decomposition XIV 7 CnHioClNOa C, H; N ¹¹ (7%) 62 2,6-Cl ₂ CeH ₃ 223,5- 225.5 I 11 C 1 H ₉ Cl ₂ NO ₃ C, H, N (0%) 63 - < 212-213 I 51 CsHuNOj C, H, N 9 64 153-155 I 3 C ₉ Hi3NO ₃ C, H, N 6.2 65 about 205,5- XIV 34 C _i0 H ₁₅ NO 3 C, N; H ^v 30 -206

SK 278319 Β6

Table I - continued

compound r ₂ mp ° C rats. solvent. ^b yield% formula analysis " K ,, μΜ or% inhibition at 100 μΜ 66 about 208- -209 I 18 C11H17NO3 C, H, N 75 67 -about 206- -207 I 18 C12H19NO3 C, H, N 6 68 205-206 I 40 C, ₅ H ₂₁ NO ₃ C, H, N (3%) 69 ^CH 2 <] 142-143 IV 9 C9H13NO3 C, H, N 22 70 -.- <> 155-156 rv 17 C10H15NO3 C, H, N 6.6 71 ^{c 2} -O 168-169 in M 2 CnH _l7 NO ₅ C, H, N 9.8 72 ^CH 2 184-185 in M 28 C12H19NO3 C, H, N (56%) 73 176-177 in M 29 C12HL9NO3 C, H, N 25 74 W 180.5 to 181 in n 10 c ₁₃ h ₂₁ no ₃ C, H, N (54%) 75 156-157 rv 10 C14H23NO3 C, H, N (68%) 76 ^{c 2} Ό 182,5- -183.5 rh, in 15 C13H19NO3 C, H, N (54%) 77 c ₂ - ^ 153-155 rv 36 C16H23NO3 C, H, N (33%) 78 ^CH 2 - 163-165 rv 18 C12H17NO3 C, H, N (50%) 79 IN] 144-145 IV 3 c ₉ h ₁₃ no ₃ H, N, C ^w (16%) 80 _With CH>< 178-180 IV 6 C9H13NO3 C, H, N 1.7 CH, F 81 - <J CH ₂ CH ₃ 167-168 in 5 C9H13NO3 C, H; N 12 82 184 IV 16 CioHjjNOj C, H, N 1.6

Table I - continued

r ₂ mp ° C rats. solvent. ^b yield% formula analysis " K i, μΜ or% inhibition at 100 μΜ , C (CH _{3) 3} 189-190 IV 45 C12H19NO5 C, H, N 6.5 - < 196 to 197.5 IN 40 C ₁₄ Hi5NO ₃ C, H, N (83%) CH ^CIJ 3 Z 142 to 143.5 ΧΠ 27 C10H15NO3 C, H, N 4 CH ™ 3 _'(+) 145.5- -146.5 ⁵ rv C ₁₀ H ₁₅ NO ₃ C, H, N 19 CM, CH, (FROM 145.5-146.5 ^y IV 1.3 r l i1 238 in n 21 C ₁₀ H ₁₅ NO ₃ C, H, N 7.2 i ch ₃ / '' 3 E., ^x ch ₃ 159-161 xn 42 ChHi ₇ NO ₃ C, H, N 7.2 CH. CH ₃ CH ₂ CH ₃ 201-202 IV 53 C12H19NO3 C, H, N (10%) 131-132 rv 12 CnHi ₇ NO ₃ C, H, N 0.45 ch ₃ CH _{(CH3) 2} * * 183-184 IV 22 C12H19NO3 C, H, N 3 = «3 / 5 <1 BP = C (CH3) ₂ 174-175 rv 18 C14H21NO5 C, H, N (15%) CH ₃ 140-142 rv 6 C14H23NO3 C, H, N 94 ^C 3 ^Cff 2 C _{(CH3) 3} CH CH 129-130 rv 10 C12H19NO3 C, H, N 0.86 Λ 189 to 190.5 xn 32 CkjHuNOs C, H, N 1.6

Table I - continued

compound R 2 mp ° C rats. solvent. ^b yield % formula analysis " Κι, μΜ or (% inhibition at 100 μΜ) 97 256 XV 8 C12H17NO3 C, H, N 4.2 98 ĹJ1 Cl 188 to 189.5 IN 18 CgHsCljNOj C, H, N, Cl 0.08 99 br 191-192 IN 23 Cgl ^ Brinch C, H, N, Br 0.03 100 F F 199-200 IN 12 C 8 H ₉ F2NO ₃ C, H, N, F. 2.1 Cl Cl 101 "X 138-141 IN 37 C ₉ Hi ₂ Cl ₂ NO ₃ C, H, N, Cl 164 102 172- -174.5 ΧΠ 19 C10H15NO3 C, H, N 4.7 103 \ z ^ ch ₃ 171-172 XVI 59 C11H17NO3 C, H, N 11.2 ^CH 3 \ / --- \ 104 XD 164-165 IV 27 CJ2H ₉ NO ₃ C, H, N (H%)

Notes to Table

I = 1,1,2,2-tetrachloroethane; Π = DMF-HOAc; m = toluene-diisopropyl ether; TV = toluene; in = CH3NO2; VI diethyl-toluene-diisopropyl ether, VL - diethyl-toluene; in M - methyl ethyl ketone; IX = acetone, H2O; x dioxane-H ₂ O; XI - diizopropvlketón; ΧΠ = EtOAc; xn = EtOH-H ₂ O; XIV = diethyl ketone; XV = metylizovalerát XVI = _CH3 CN.

The analyzes had a deviation of ± 0.4% of the theoretical value unless otherwise stated.

^d Melting point reported in the literature is 159-160 ° C [VE Price and JP Greenstein, Ar. Biochem., 18, 383 (1948)] ^e H: calcd 9.31; found 9.91 ^f H: calculated 11.24; found 11.84 ⁸ H: calculated 9.54; found 10.07 ^h C: calculated 61.53, found 61.07 ¹ HE Carter, P. Handier and DB Melwille, J. Biol. Chem., 129,359 (1939) ^J Trichlorethylene as solvent ^k The total yield for grade ¹ admissions. Melting point 170-172 ° C [M. Flavin and C. Slaughter, J. Biol. Chem., 244, 5, 1434 (1969)].

¹¹¹ p-toluenesulfonic acid catalyst in the literature ⁿ declared melting point is above 270 ° C (Flavin M. Slaughter, C., loc. Cit.) ⁰ The product was obtained by concentration of the mixture and rozotre10 the residue with ethanol to a solid.

^p The product was obtained by decantation of the solvent, extraction of the residual oil with ethyl acetate and evaporation of the extract the crystals.

⁴ The product was obtained by decantation of the solvent and trituration of the residual oil in diethyl ketone to a solid.

¹ Softening above 90 ° C.

⁵ H: Calcd. 9.29; found 9.74 ¹ C: calculated 61.27; found 60.74 ¹¹ N: calculated 5.85; found 6.40 ⁱⁿ H: calc. 7.67; found 8.14 ^w C: calculated 59.00; found 58.49 ^x [α] D + + 98.5 ° (c 0.35, CHCl 3) γ [α] ²⁴ . 97 _{> 2} »( _C 0.35, CHCl 3) ^z Methylisovalerate as solvent.

Π (Z) -3-Substituted 2- (2,2-dimethylcyclopropanecarboxainido) propanoic acid

H CO.H

W

CH ₃ CH ₃

compound Rs A configuration mp ° C rats. solvent ^b yield% formula analysis Ki, μΜ 105 H RS 122-123 I 5 C ₉ H ₁₃ NO 3 C, H, N 75 106 CH3CH2 RS 154,5- Π 30 CnHnNOj C, H, N 0.18 107 CH ₃ (CH ₂ ) 2 RS 155.5 122-123 m 10 C12H19NO3 C, H, N 0.11 108 (CH ₃ ) ₂ CH RS 164-166 1 27 C ₁₃ H ₂ iNO ₃ C, H, N 0.54 109 CH ₃ (CH ₂ ) ₃ RS 94-95 IV 24 c ₁₃ h ₂₁ no ₃ C, H, N 0.11 110 (CH ₃ ) ₂ CHCH 2 RS 145-147 XXIV 35 CuH2iNO ₃ C, H, N 0.23 111 CH ₃ (CH ₂ ) ₄ RS 104-105 I 10 C14H2JNO3 C, H, N 0.17 112 113 114 CH ₃ (CH _{2) 4} CH ₃ (CH _{2) 4} (CH _{3) 2} CH _{(CH2) 2} R ^with R-RS 95 to 96.5 94,5- -96.5 115-116 in 20 C14 H ₂₃ NO ₃ C14 H ₂₃ NO ₃ C, H, N C, H, N 0.80 8.80 0.15 115 (CH ₃ ) ₃ CCH 2 RS 138,5- IV 28 c ₁₄ h ₂₃ no ₃ C, H, N 0.34 116 CH ₃ rcn ₂ ) 5 RS 140 111-113 I 11 C ₅ H ₂₅ NO ₃ H, N; C ~ 0.16 117 - (CH ₂ ) ₆ - RS 188-190 VI 9 C 24 H ₃₆ N 2 O ₆ .1 / 2H ₂ O C, H; N ^f 0,092 118 CH ₃ (CH ₂ ) ₆ RS 108-110 I 13 CI _N H ₃₇ NO ₃ C, H, N 0,096 119 CH ₃ (CH ₂ ) ₇ RS 95-96 in n 47 C ₁₇ H2 ₉ NO ₃ C, H, N 0.11 120 CH ₃ (CH 2) 8 RS 91-92 IV 52 CISH ₃ iNO ₃ C, H, N 0.11 121 CH ₃ (CH 2) 9 RS 98-99 in n 25 C ₉ H ₃₃ NO ₃ C, H, N 0.14 122 CH ₂ = CH (CH ₂ ) 2 o- RS 88-90 n 39 C ₃ H ₃ NO _h C, H, N 0.23 123 RS 158-159 n 4+ Ci ₂ Hi ₇ NO ₃ C, H, N 0.44 124 0 RS 149- H 40 Ci5H ₂₃ NO ₃ C, H, N 0.40 125 w u RS 150.5 146-148 n 31 ο _{] 6 and 25 3} C, H, N 0.15 126 127 cf ₃ ch ₂ CH ₃ O (CH 2) 2 S RS 139-142 g I 19 C12H15NO4 C, H, N C, H, N 0.24 0.32 128 CH ₃ S (CH ₂ ) ₂ RS 65-67 VNI 5 Ci ₂ Hi ₉ NO ₃ S C, H, N 0.12 129 130 CH ₃ SO ₂ (CH ₂ ) ₂ HO ₂ C (CH ₂ ) ₂ RS RS 148-149 163-165 VI 8 CnHpNOsS C12H17NO5 C, H, N, S; C, H, N 0.50 0.14 131 Br _{(CH2) 3} RS 118-120 I 5 C ₁₂ Hi8BrNO ₃ C, H, N 0.38 132 CH ₃ O (CH ₂ ) ₃ RS 78-80 in M 33 C13H21NO4 C, H, N 0.28 133 CH ₃ O ₂ CCH ₂ S (CH ₂ ) ₃ RS 133-135 XXM 71 C15H23NO5S C, H, N 0.28 134 L-HO ₂ CCl I (NH ₂ ) CH ₂ S (CH ₂ ) ₃ RS 120-132 zaroz- IX 68 C ₁₃ H ₂₄ N ₂ O ₅ S.1 / 2H ₂ O C, H, N, S; 0.27 135 γυ ^οκ RS kladu g 40 C ₁₇ H2 ₂ N ₂ O ₄ S.1 / 4H ₂ O C, H, N 0.13 136 137 Na 2 HO ₃ PCH ₂ S (CH ₂ ) ₃ HO ₂ C (CH 2) ₃ RS RS g 114-115 x 18 C ₁₃ H ₉ N ₃ O ₂ PS · 1 / 3H 2 O C ₁₃ Hi ₉ NO ₅ C, H, N C, H, N 0.22 0.48

SK 278319 Β6

Table Π - continued (Z) -3-substituted 2- (2,2-dimethylcyclopropanecarboxamido) -propanoic acid

compound Rs configuration melting point ° C rats. solvent ^b yield% formula analysis 138 Br _{(CH2) 4} RS 124-125 I 36 C13H20B1NO3 C, H, N 0.15 139 Br _{(CH2) 4} S ¹ 86-88 I 28 C 15 H _{2 and} BrNO ₃ C, H, N, Br 140 CH ₃ O (CH ₂ ) ₄ RS 102-104 m 35 C14H23NO4 C, H, N 0.16 141 NC _{(CH2) 4} RS 150 XI 46 C 14 H ₂₀ N ₂ O ₃ C, H, N 0.21 142 (CH ₃ ) ₂ N (CH ₂ ) ₄ RS 158-161 XVII 7k Ci5H N _{2 S 2} O ₃ .l / 4H ₂ O C, H, N 3.45 143 Na ⁺ O ₃ S (CH ₂ ) ₄ RS g CiÄNNaOsS. C, H, N 0,087 ., 3 / 4H ₂ O 144 HO ₂ CCH ₂ N (CH ₃ ) (CH ₂ ) ₄ RS g and 12 C16H ₂ 6 N ₂ O5. .1 / 2AcOH.2 / 5 H ₂ O C, H, N 0.56 145 HO ₂ CCH ₂ S (CH ₂ ) ₄ RS 119-121 I 91 Ci5H ₂₃ Noss C, H, 0.13 N, S 146 L-HO ₂ CCH (NH ₂ ) CH ₂ S (CH ₂ ) ₄ RS g and 71 Ci6H N _{2 S 2} O ₅ Sl / 4 H ₂ O C, H, 0.21 N, S 147 D-HO ₂ CCH (NH ₂ ) CH ₂ S (Cl ₂ ) ₄ RS g and 75 CisHjsNíOsS.IÄIHíO C, H, 0.19 N, S 148 L-HO ₂ CCH (NH ₂ ) CH ₂ S (CH ₂ ) ₄ WITH- g and 63 C16H26N2O5S C, H, 0.11 N, S 149 L - ^ C ₂ CCH (NHCOCH ₃ ) ch ₂ s (ch ₂ ) ₄ S ¹ g IX 68 Ci8H ₂ 7 N ₂ NaO ₆ SH ₂ O C, H, N, S; 0.07 150 L-HO ₂ CCH _(NHCH3) s ^ra 144-148 IX 54 C17H28N2O5S.I / C, H, 0.15 CH ₂ S (CH _{2) 4} Z2H ₂ O N, S 151 HO ₂ CCOH ₂ S (CH ₂ ) ₄ RS n 78 c ₁₆ h ₂₃ no ₆ sec./ C, H, N 0.16 / 3H ₂ O 152 C ₆ H ₅ S (CH ₂ ) ₄ RS 100-104 I C ₁₉ H ₂₅ NO ₃ C, H, N 0.10 / 4H ₂ O 153 2-CO ₂ HC ₆ H ₄ S (CH ₂ ) ₄ RS 163-167 I 21 C ₂ o ₂₅ N ₅ S.3 / C, H, N 0.09 / 4H ₂ O C, H, 154 í 1 S ° 188,5- ΧΠ 65 C ₂₄ H ₂₄ N ₂ O ₄ S N, S 0.02 189 155 Q WITH g 50 C ₁₉ H ₂₄ N ₂ O ₅ P.1 / 4H ₂ O C, H, N 0.04 156 HO ₂ C (CH ₂ ) ₄ RS 165-167 VI 28 C ₄ H ₂ INNO ₅ C, H, N 0,058 157 HO _{(CH2) 4} RS 137-139 C ₁₄ H ₂₃ NO ₄ C, H, N 0.23 158 CH ₃ CO ₂ (CH ₂ ) ₃ RS 59-62 CiíHjsNOs C, H, N 0.22 159 CH ₃ O (CH ₂ ) ₅ RS 71-72 C ₁₅ H ₂₅ NO ₄ C, H, N 0.18 160 Br _{(CH2) 3} RS 149-151 XIV 55 C ₁₄ H ₂₂ BRNO ₃ C, H, 0.27 N, Br 161 Br _{(CH2) 5} S ^E 100,5- I 44 CiÄíBrNOs C, H, -101.5 N, Br 162 NC (CH ₂ ) ₃ RS 113-115 XVI 38 Ci5H ₂₂ N ₂ O ₃ .l / 4H ₂ O C, H, N 0,082 163 H ₂ N (CH ₂ ) ₃ RS g XVN 78 C14H24N2O3.H2O C, H, N 1.00 164 (CH ₃ ) ₂ N (CH ₂ ) ₅ RS 101-102 XIX 71 ClíH ^ Oj. H ₂ O C, H, N 1.28 165 (C ₂ H ₅ ) ₂ N (CH ₂ ) ₅ RS 78-81 XV 77 C ₁₈ H ₃₂ N ₂ O ₃ C, H, N 0,086 166 RS 126-129 Ci ₇ H ₂₈ N ₄ O ₃ C, H, N 0.84 167 1 H HO ₂ CCH ₂ N (CH ₃ ) (CH ₂ ) ₅ RS g XVN 27 C17H28N2O5.2H2O C, H, N 0.29 168 (HO) 2P (O) CH ₂ NH (CH ₂ ) ₃ RS 125-130 and 33 C1 ₅ H ₂ 7N ₂ O ₆ P.H ₂ O C, H, 0.40 N, P 169 (HO) 2P (O) (CH ₂ ) ₂ NH (CH ₂ ) j RS g and 96 Ci6H ₂₉ N ₂ O ₂ O6P.lZ2H C, H, 0.58 N, P 170 D, L- (HO) ₂ P (O) CH (CH ₃₎ RS 136-139 and 29 CléH ^ osp C, H, 0.28 NH (CH ₂ ) _s N, P

Table Π - continued

compound r ₃ configuration mp ° C rats. solvent. ^b yield% formula analysis ⁰ Ki, μΜ 171 D, L- (HO) 2 P (O) CH (CH 3) NH (CH ₂ ) ₅ s ^ä 143-145 and 54 CAW C, H, N 0.16 172 (HO) ₂ P (O) C (CH ₃ ) ₂ NH (CH ₂ ) ₅ s ¹ 162-165 and 49 C ₁₇ H ₃ N ₂ O ₆ P C, H, N, P 0.18 173 CH ₂ -CHCH ₂ NH (CH ₂ ) ₅ RS 163- 165 164- 166 XVN 92 C17H2 & N2O3. C2H5OH H, N; b 0.87 174 HCl-CH ₂ NH (CH ₂ ) ₅ RS RS with decomposition 220-222 XVM 66 C17 ₂ 6 N ₂ O ₃ C, H, N 0.74 175 (CH ₃ ) ₃ N (CH ₂ ) ₅ decomposition 225-227 XIX 77 C17H30N2O3 C, H, N 1.10 176 (CH ₃ ) ₃ n (CH ₂ ) ₅ S ^! for decomposition XIX 71 Ci7ÍI ₃ cN ₂ O ₂ C, H, N 0.65 177 (HOCH ₂ CH ₂ ) (CH ₃ ) ₂ N (CH ₂ ) 5 RS 185-188 XX 55 C18H32N2O4.1- 1/4 H ₂ O C, H, N 0.91 178 ς \ (ch ₂ ) ₅ RS g and 4 C19H32N2O3.2H2O H, N, E ^N 1.09 179 180 CH ₃ CONH (CH ₂ ) with H ₂ NCONH (CH ₂ ) ₅ RS RS g g 160-162 XXI 79 C _1S H ₂₆ N ₂ O ₄ . .1 / 5CH3COCH3 CisHÄCU, 2 / 3H ₂ O C15H25N3O3. C, H, N C, H, N 0.23 0.30 181 H ₂ NHC = N (CH ₂ ) ₅ RS for decomposition XVN 77 , 1 / 3H ₂ O C, H, N 0.78 182 183 184 H ₂ NC (CH ₃ ) = N (CH ₂ ) ₅ CH ₃ CH ₂ O ₂ CS (CH ₂ ) ₅ HS (CH ₂ ) ₅ RS RS RS 143-145 88-91 128-130 XX 60 C16H27N3O3. , 3 / 4H ₂ O C ₂ H ₂ N 2 CHS C 14 H 23 NO 3 S C, H, N C, H, N, S; C, H, N, S; 0.72 0,044 0.17 185 CH ₃ S (CH ₂ ) ₅ RS 82-84 XI 60 C ₁₅ H ₂₅ NO ₃ S C, H, N, S; 0.15 186 CH ₃ O ₂ CCH ₂ S (CH ₂ ) 5 RS g xm 60 C17H27NO4S C, H, N, S; 0.20 187 H ₂ NCOCH ₂ S (CH ₂ ) ₅ RS 125-126 174-177 VI 55 C ₆ H 6 N ₂ O ₄ S ₂ H, N, S; C ² C, H, 0.25 188 L-HO ₂ CCH (NH ₂ ) CH ₂ S (CH ₂ ) 5 RS for decomposition and 54 Ci7H ₂ gN20sS. , 1 / 3H ₂ O C ₇ H ₂ 6N ₃ O ₂ S ₂ . N, S C, H, 0.23 189 RS 86-90 I 4 .036CH3NO2 N, S 0.15 190 Br _{(CH2) 6} RS 129-131 I 57 C ₂ H ₅ 4BrNO3 C, H, W, Br 0.16 191 HO ₂ CCH ₂ N (CH ₃ ) (CH ₂ ) ₆ RS g XVN 51 C 18 H 30 N ₂ C> 5. , 1 / 2H ₂ O C, H, N 0.40 192 Br (CH _{2) 7} RS 96-97 IV 71 Ciofu ^ palsy? C, H, N, Br 0.11 193 H ₂ N (CH ₂ ) ₇ RS 110-120 ΧΧΠ 58 C ₁₅ H ₂ xN ₂ O ₃ C, H, N 0.81 194 (CH ₃ ) ₂ N (CH ₂ ) ₇ RS 168-172 IV 43 C18H3 ₂ N ₂ O _3rd .1 / 2H2O C, H, N 0.52 195 (CH ₃ ) ₃ N (CH ₂ ) ₇ + RS RS g 190-193 XV 29 C ₁₉ H34N ₂ O3.H ₂ O C, H, N 0.57 196 (C 5 H 11 CH ₂ XCH 2 N (CH ₂ ) ₇ ) for decomposition XVN 83 C ₂₅ H 38 N ₂ O 3.1 / 2H ₂ O H, N; C- 0.45

SK 278319 Β6

Table Π - continued

compound r ₃ configuration mp ° C rats. solvent. ^b yield% formula analysis ⁰ Ki, μΜ 197 CôHs RS 167-168 VI 17 C ₁₅ H 17 NO ₃ C, H, N 0.62 198 C6H5 (CH2) ₂ RS 131-132 I 33 C ₇ H ₂ iNO ₃ C, H, N 0.33

Notes on Table Π ^b I = CH3NO2, H = toluene, IH = toluene-diisopropyl ether, 1V = ether petroleum ether, V = toluene-cyclohexane, VI = EtOAc, VII = hexane, VHI = ether-cyclohexane, IX = = MeOH- ether, X = EtOAc-hexane, XI = ether-hexane, XH = CH ₃ NO ₂ -acetic acid, XH = MeOH-chloroform, XTV = CH ₃ CN, XV = EtOH-ether-acetone, XVI = H ₂ O, XVĽ = EtOH-H ₂ O, XVm = = tetrahydrofuran-hexane, XIX = EtOH-acetone, XX = = MeOH-H 2 O, XXI = acetone-H ₂ O, XXH = acetone ether, XXHI = petroleum ether, XXIV = this diisopropyl ketone.

'[a] I "+ 77.7 ° (H, 51, CHC1 ₃₎ ^d [a] ¹ g of -78.0 ° (0.48, CHC1 ₃₎ ^and C: calcd, 67.38; found 66.88 ^f B: calculated 6.12; found 5.71 ^g of an amorphous solid, m.p. slurred Metylizovalerát ^h as the reaction solvent 1 ^ + 72,8- (0.5, CHCl,) ^J Purification by ion exchange ^of [a] + 17.6 ° (0.5 , MeOH), + 14.2 ° (0.5, 0.1 N, GC1) '[a] D-9.8 ° (0.5, 0.1 NNaOH) ^5' [α] 35 + β. 0 ° (0.5, 0.1 NHC 1) ⁿ 01ej. Mixture of enol isomers ° [α] B7 + 34,3 ° (0,5, CH 3 OH) ^P []] _K + 66,2 ° (0,5, CHCl ₃ )

W + 12.5 ° (0.6, H ₂ O)

Mp + 30.5 ° (0.24, H ₂ O)

U ^S C: calcd 64.37, found 64.96 * [α] l ⁵ + 58.0 DEG (0.5, _CH3 OH) ^The C: calcd 61.26; found 61.73 ^V C: calc. 56.11; found 55.61 ^W C: calc. 70.89; found, 7.34.

Table ΙΠ

3-substituted 2- (acylamino) propenoic acid

compound R 2 r ₃ R4 mp ° C rats. solvent ^b formula analysis K i, μΜ or% inhibition at 100 μΜ 199 200 201 202 Hochjoch (CH,) ₇ CHCH 2 (CH _{3) 2} CHCH 2 (CH _{3) 2} CHCH ₂ CH ₃ CH ₂ CH ₃ (CH ₂ ) 4 HO ₂ C (CH ₂ ) 2 H H H H 172-173 128.5 to 129.5 126-128 127-128 C hl I CnHnNCU CioHi ₇ N0 ₃ C13H ₂₃ NO 3 NO _{7 5} CHHINE C, H, N C, H, N (4%) 3.2 2.6 2.7 203 l> - HO ₂ C (CH 2) 2 H 119-120 n ChoHisNOj C, H, N 5.0 204 ý Cl Cl CH ₃ (CH ₂ ) 4 H 135 to 136.5 m C ₂ Hi ₇ Cl ₂ NO ₃ C, H, N, Cl 0.06 205 Cl Cl Br _{(CH2) 5} H 159 to 160.5 hl CuHieBr Cl ₂ NO ₃ C, H, N, Cl B / 206 J Cl Cl (CH ₃ ) ₃ N (CH ₂ ) ₅ H 183-185 with decomposition ¹ rv C15H ₂ 4C1 ₂ N ₂ O ₃ .C ₂ H ₅ OH C, H, N 0.18

Table H1 - continued

compound R 2 Rs R4 melting point ° C rats. solvent ¹¹ formula analysis K i, pM or% inhibition at 100 pM 207 br ^ br CH ₃ (CH ₂ ) 2 H 178-179 m C ₁₀ Hi3 Br ₂ NO ₃ C, H, N, Br 0,015 208 CSH, H H 125-127® (9%) 209 c ₆ h ₅ Cl H h (36%) 210 qa CH3O H 216-217 za (23%) decomposition 214 L-CH NH _o ⁿ | £ Cl H ABOUT CsHsCINíOj. C, H, N (6%) ch ₃ 2 / 3H ₂ O

Notes to Table Hl

Ή = EtOAc, Π = H _2, H = CII3NO2, IV from EtOH-ether, ⁰ is purified by chromatography solvent metylizovalerát ^{d e} p-toluenesulfonic acid as the catalyst ^F 5 Br: Calcd 21.42; found 20.92 ^g Melting point 122 ° C (dec.) [T. Wieland, G. Ohnacker and W. Ziegler, Chem. Ber., 90, 194 (1957)] ^h Strukov, Zh, Obshch. Khim., 27, 432 (1957) 10 ^I. Melting point 203-204 ° C (dec.) [W. Schulz, Chem. Ber., 86.1010, (1953)] ^II [α] ²⁴ + 25.3 ° (2.1 N HCl) ⁰ amorphous solid. 15

Claims (5)

PATENT CLAIMS (Z) -2-Acylamino-3-monosubstituted propenoates of general formula (I) (D) R ¹ represents a hydrogen atom, a C 1-6 alkyl group, a di (C 1-2 alkyl) amino (C 1-3 alkyl) group, or a pharmaceutically acceptable cation; R ² is a hydrocarbon radical having 3 to 10 carbon atoms in which the carbon chain may be from 1 to 6 hydrogen atoms are replaced by halogen atoms, and a methylene group in a non-terminal position can be replaced by oxygen or sulfur, including oxidized forms of sulfur; R ³ represents a hydrocarbon radical having 1 to 15 carbon atoms, in which the carbon chain may be replaced by 1 to 6 carbon atoms by halogen atoms, the methylene group other than the terminal position may be replaced by an oxygen or sulfur atom, including oxidized sulfur, and a terminal atom the hydrogen may be replaced by a hydroxyl or thiol group, or an amino group which is optionally derivatized in the form of an acylamino group having 1 to 2 carbon atoms, an ureido group, an amidine group, a guanidine group, a monoalkylamine group, a dialkylamine and a trialkylamine group an alkyl moiety, a carboxy group, a group of formula (HO / 2P / O) - or -SO 3 OH, or a corresponding C 1 -C 2 alkyl ester or alkylamide group derived from any of these acidic groups, cyano or combinations thereof, such as terminal residues and m and when R ³ is C 1 -C 4 straight chain alkyl, R ^{2 is not} C 1 -C 4 alkyl. Propenoates according to claim 1, wherein R ² represents 2,2-dimethycyclopropyl and R ³ represents a group of the formula -CE ₂ -3-OH ₂ -CH-COO ^_ A process for the preparation of propenoates of formula (I), wherein R ¹ is hydrogen, C 1 -C 6 alkyl or a pharmaceutically acceptable cation and R ² and R ³ are as defined in claim 1, according to claim 1, characterized by: by weight of 2 parts of the 2-keto acid of the formula The R ³ CH ₂ --C ₂ h wherein R ³ is as defined in Claim 1, or an alkyl ester having 1 to 6 carbon atoms in the alkyl radical condensed with 1 part of an amide of the above Formula J R ^Z CHH ₂ Wherein R ² is as defined in claim 1, by refluxing the reaction mixture in an inert solvent such as toluene or methylisovalerate while simultaneously azeotropically separating water, then optionally forming the free acid of formula (I) to its 1 to 1 alkyl ester. Or the resulting ester is converted to the free acid, or the resulting product is converted to its pharmaceutically acceptable salt. A process according to claim 3, for producing propenoates of formula (I), wherein R ¹ is as defined in Claim 3, ^R2 is 2,2-dimethylcyclopropyl or a 2,2dichlórcyklopropyl R ³ is a hydrocarbon radical having 3 to 7 carbon atoms, optionally substituted on the terminal carbon trimethylammonium, amidino, guanidine, or 2-amino-2-carboxyethylthio, characterized in that the amide of formula wherein R ³ is as defined above, in the presence of a base followed by oxidative addition of sodium methoxide, and the resulting product is treated with 5 anhydrous hydrochloric acid, then the resulting product is optionally converted to its pharmaceutically acceptable acid addition salt or other ester within the definition of R ¹ . Propenoates of formula (I) according to claim 10 1 for use as dipeptidase inhibitors. An antibacterial preparation based on an antibiotic of the thienamycin family, such as thienamycin, N 15 -formimidoylthienamycin and N-acetylimidoylthienamycin, characterized in that it comprises a combination of the antibiotic with a dipeptidase inhibitor propenoate of formula (I) according to claim 1 in a weight ratio of the first substance. to the second substance in 20 ranging from 1: 3 to 30. First The end of the document where R ² is as defined above is reacted with a keto acid of formula O r ³ -ch ₂ -c-cooh where R ³ is a hydrocarbon radical having 3 to 7 carbon atoms, the terminal carbon is replaced with a bromine atom, then the resulting bromo intermediate is reacted with trimethylamine, with a mixture of ammonium salt and imide, with a guanidine or cysteine hydrochloride. Process for the preparation of the propenoates of the formula I according to claim 1, characterized in that the acid chloride of the formula I is reacted. O J Wherein R ² is as defined above, with the t-butyl ester of the α-amino acid of the general formula H R ³ -CH ₂ -C-COO-C (CH ₃ ) -j kh ₂