IE47081B1 - -halomethyl derivatives of -amino acids - Google Patents

Abstract

Thermoplastic polypropylene elastomer blends and a partially crosslinked ethylene-vinyl acetate copolymer. Mixtures of polypropylene and ethylene-vinyl acetate copolymers. Mixtures of polypropylene and copolymers of ethylene-vinyl acetate without crosslinking, have unsatisfactory elastic properties, for example, with respect to permanent elongation and impact resistance in notched piece. It has been found that by using partially crosslinked ethylene-vinyl acetate copolymers with gel contents of 3-50% by weight in admixture with isotactic polypropylene, improvements in elastic properties are achieved, especially in part impact resistance notched and in permanent elongation, with respect to comparative mixtures with copolymers of ethylene-vinyl acetate without crosslinking.

Description

This invention relates to novel and useful halomethyl derivatives of α-amino acids.

The compounds of the present invention may be represented by the following general Formuia I: Y I Z-^-CORi Formula I NHRb In the above general Formula I Y is FCH2-, F2CH-, F3C-, CICH2-, or Cl2CH-; Z is β-methylthioethyl, β-thioethyl, β-benzylthioethyl, S-(5'-desoxyadenosin-5'-yl)-p-methylthioethyl, γ-guanidinopropyi, or RaHN(CH2)n- wherein n is Ya the integer 3 or A or HOOC-C-(CH2)2-S-S-(CH2)s- wherein NH2 Y2 is FCH2-, F2CH-, or FgC; each of Rg and Rb is the same or different and is hydrogen, alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and is straight or branched, alkoxycarbonyi wherein the alkoxy moiety has from 1 to 4 carbon atoms and is straight or branched, or II the group -C-CH-R2 wherein R2 is hydrogen, a straight or NH2 branched lower alkyi group of from 1 to 4 carbon atoms, .benzyl or p-hydroxybenzyl; and Rt is hydroxy, a straight or branched alkoxy group of from 1 to 8 carbon atoms, -NR4Rs wherein each R4 and Rs is hydrogen or a lower alkyl group of from 1 to 4 carbon atoms, or the group -NH-CH-COOH Rs wherein R3 is hydrogen, a straight or branched lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzyl; with the provisos that when Z is γ-guanidinopropyl or β-methy1thioethyl, Y is FCH2-, F2CH-, or F3C7 when Z is γ-guanidinopropyl Ri is hydroxy; and when Z is β-thioethyl, β-benzylthioethyl, S-(51-desoxyadenos in-51-yl)-S-methy1Ϊ8 thioethyl or HOOC-C-(CH2)2-S-S-(CH2)2-, Y and Y2 are FCH2-.

NH2 F2CH-, or F3C- and are the same, each of Rg and R^ is hydrogen and Rt is hydroxy. The lactams of the compounds of general Formula i wherein L is RaHN(CH2)n- and each of Rg and R& is hydrogen are also within the scope of the present invention. Pharmaceutically acceptable salts and individual optical isomers of the compounds of general Formula I are also included within the scope of the present invention.

DETAILED DESCRIPTION OF INVENTION In the above general Formula I in addition to the YE group RaHN(CH2)n- and HOOC-C-(CK2)2-S-S-(CH2)2- the symbol NHa Z represents the substituent groups β-methylthioethyl, βthioethy), β-benzylthloethyl, S-(5'-desoxyadenosin-51-yl)β-methylthioethyl and γ-guanidinopropyl which are depicted by the following structures: HaC-S-CHsCHs HS-CH2CH2Q-ch2-s-ch2ch2β-methylthioethyl β-thioethyl β-benzylthioethyl S-(5'-desoxyadenos in5-yl)-S-methyl thioethyl HN II , , H2N-C-NH(CH2)3y-guan id 1 nopropyl As used in general Formula | the term alkylcarbonyl is taken to mean the group -C-alkyl wherein the alkyl moiety has From 1 to 4 carbon atoms and is straight or branched, for example, methyl, ethyl, ri-propyl, isopropyl £-butyI, isobutyl and tert-butyl.

As used in general Formula 1 the term alkoxycarbonyl II is taken to mean the group -C-O-alkyl wherein the alkoxy moiety, that is, -0-alkyl, has from 1 to 4 carbon atoms and is straight or branched, for example, methoxy, ethoxy ri-propoxy, isopropoxy, jn-butoxy, and tert-butoxy.

Illustrative examples of straight or branched alkyl groups having from 1 to 4 carbon atoms as used in general Formula I are methyl, ethyl, £-propyl, rt-butyl, isopropyl and tert-butyl.

Illustrative examples of alkoxy groups having from 1 to 8 carbon atoms as used in general Formula I are methoxy, ethoxy, propoxy, butoxy, pentyloxy, and octyloxy The lactams of the compounds of general Formula 1 wherein Z is R,HN(CH2) -, and each of R, and R. is hydro3 Π a D gen are represented by the following general Formula II: 7 0 8 1 nh8 Y-C-C=0 Formula II I I (CH2) —NH In the above general Formula II, n is the integer 3 or 4, and Y is FCH2-, F2CH-, F3C-, C1CH2- or C12CH-.

Illustrative examples of pharmaceutically acceptable salts of the compounds of this invention include non-toxic acid addition salts formed with inorganic acids, such as, hydrochloric, hydrobromic, sulfuric and phosphoric acid, and organic acids, such as, methane sulfonic, salicylic, maleic, malonic, tartaric, citric, cyclamic and ascorbic acids; and non-toxic salts formed with inorganic or organic bases such as those of alkali metals, for example, sodium, potassium and lithium, alkaline earth metals, for example, calcium and magnesium, light metals of Group 111 A, for example, alumi-num, organic amines, such as, primary, secondary or tertiary amines, for example, cyclohexylamine, ethylamine, pyridine, methylaminoethanol, ethanolamine and piperazine. The salts are prepared by conventional means.

Preferred compounds of this invention are those of general Formula I wherein Ri is hydroxy. More preferred compounds of this invention are those of general Formula 1 wherein Ri is hydroxy, Z is β-methylthioethyl, S-(5'desoxyadenosin-51-yl)-S-methylthioethyl, γ-guanidinopropyl or RaHN(CH2)n- and each of Rg and R^ is hydrogen and the lactams of said compounds wherein Z is RaHN(CH2)n~.

Another preferred embodiment of this invention is the compounds of general Formula I and the lactams thereof wherein Y is other than F3C- with compounds wherein Y is FCH2- or F2CH- being more preferred.

Illustrative examples of compounds of the present invention are the following: 2-amino-2-fluoromethyl-4-methylthiobutyric acid, 2-amino-2-d ifluoromethyl-4-methylthiobutyric ac id, 2-amino-2-trifluoromethyl-4-[S-(5'-desoxyadenos in-5'-yi)S-(methyl)thiojbutyric acid, 2-amino-2-fIuoromethyi-4-thiobutyric ac id, 2-amino-2-difluoromethyl-4-thiobutyric acid, 2-amino-2-fluoromethyl-5-guan id inovaler ic ac id, 2-dichloromethyl-2,5-diaminovaleric acid, 2-dichloromethy1-2,6-diaminocaproic acid, methyl 2-amino-2-difluoromethyl-4-methylthiobutyrate, 2-amino-2-tr ifluoromethyl-4-methylth iobutyr ic acid, 2-amino-2-difluoromethyl-4-[S-(5'-desoxyadenos in-5'-yl)-S (methyl)thio]butyric acid, 2-amino-2-fluoromethyl-4-[S-(5'-desoxyadenosin-5'-yl)-S(methyl)thio]butyric acid, 2-amino-2-difluoromethyl-5-guan id inovaler ic acid, 2-amino-2-trifluoromethyl-5-guanidinovaleric acid, 2,5-diamino-2-chloromethylvaleric acid, 2.5- diamino-2-f1uoromethy1valeric acid, 2.5- diamino-2-difluoromethylvaleric acid, 2.5- diamino-2-trifluoromethylvaleric acid, 2.6- diamino-2-chloromethylcaproic ac id, 2,6-diamino-2-fluoromethylcaproic acid, 2.6- d i am i no-2-d i fIuoromethylca pro i c acid, 2,6-d i am i no-2-tr i f1uorome thylcaproic acid, 2-amino-4-benzylthio-2-difluoromethylbutyric acid, 2,9-d i am i no-2,9-d i f1uo romet h y1homoc ys t i ne, 4-70 81 methyl 2-amino-2-d i fluoromethyl-4-[S-(5'-desoxyadenos in5' -yl)-5-(methyl)thio]butyrate, methyl 2-amino-2-fluoromethyl-5-guanid inovalerate, methyl 2,5-diamino-2-d ifluoromethylvalerate, methyl 2,6-diamino-2-chloromethylcaproate, ethyl 2-amino-2-fluoromethyl-4-methy1thiobutyrate, ethyl 2,5-diamino-2-dichloromethylvalerate, £-propyl 2-amino-2-fluoromethyl-5-[S-(5'-desoxyadenos in5'-yl)-S-(methyl)thio]butyrate, £-propyl 2,5-diamino-2-tr ifluoromethyIvalerate, £-butyl 2,6-diamino-2-d i fluoromethy1caproate, isopropyl 2-amino-2-di fluoromethyl-4-methyIth iobutyrate, hexyl 2,5-diamino-2-chloromethylvalerate, Ν,Ν-dimethyl 2,5-diamino-2-difluoromethyIva 1 eramide, isopropyl 2,5-diamino-2-f1uoromethylvalerate, piva1yl 2,6-diamino-2-trifluoromethyIcaproate, N-methyl 2-amino-2-difluoromethyl-4-methylthiobutyramide, N-methyl 2-amino-2-trif Iuoromethyl-4-[5-(51-desoxyadenos in5'-yl)-5-(methyl)thiojbutyramide, N-ethyl 2-amino-2-di fluoromethyl-5-guan id inova Ieramide, N-£-butyl 2,5-diamino-2-trif 1uoromethylvaleramide, 2-amino-2-trif 1uoromethyl-4-methy1thio-1-oxobutylaminoacetic acid, 2-(2-amino-2-chloromethyI-1-oxo-l,5-pentylenediamine)pheny1propionic acid, 6-ami no-2-dichloromethyl-2-(l-oxopropylamino)caproic acid, 2-d ifluoromethy1-4-[S-(5'-desoxyadenos in)5'-yl)-S-(methyl )thio]-2-(l-oxoethy1amino)butyr ic acid, ethyl 2-fluoromethy1-5-guan idino-2-(1-oxopropylamino)valerate, '» 4 7081 N-methyl 2-dichloromethyl-2,5-di-(l-oxoethy1amino)valeramide, and methyl 2-difluoromethyI-2,5-di-(2-amino-loxoethy1amino)caproate.

The compounds of general Formula I have many utilities The compound of general Formula I wherein Z is β-thioethyl, Y2 β-benzylthioethyl or H00C-C-(CH2)2-S-S-(CH2)2-, each of R I nh2 and Rb is hydrogen and R! is hydroxy are useful as intermediates in the preparation of the corresponding pharmaceutically useful compound wherein Z is S-(5’-desoxyadeno10 sin-5’-y1)-S-methyithioethyl.

The compounds of general Formulas I and II wherein Z is other than β-thioethyl, β-benzylthioethyl or Ya HOOC-C-(CH2)2-S-S-(CH2)2- are inhibitors of decarboxylase NH2 enzymes which are involved in polyamine formation render15 ing said compounds useful as pharmacological agents. Polyamines, particularly putrescine, spermidine and spermine are present in plant and animal tissues and in some microorganisms. Although the exact physiological role of polyamines has not been clearly delineated there is evi20 dence to suggest that polyamines are involved with ceil division and growth. (H.G. WiiIiams-Ashman et al., The Italian J. Biochem. 25, 5-32 (1976), A. Raina and J. Janne, Med. Biol. 53, 121-147 (1975) and D.H. Russell, Life Sciences 13, 1635-1647 (1973))- Polyamines are essential growth factors for or involved in the growth processes of certain microorganisms, for example, E. coli, Enterobacter, 47β8ί Klebs iella, Staphylococcus aureus, C. cadaver is, Salmonella typhosa and Haemophilus parainfluenza. Polyamines are associated with both normal and neoplastic rapid growth there being an increase in the synthesis and accumulation of polyamines following a stimulus causing cellular proliferation. Also, levels of polyamines are known to be high in embryonic systems, the testes, in patients with rapidly growing tissues. It is known that there is a correlation between the activity of the decarboxylase enzymes of ornithine, S-adenosylmethionine, arginine and lysine and polyamine formation.

The biosyntheses of putrescine, spermidine and spermine are interrelated. Putrescine is the decarboxylation product of ornithine, catalyzed by ornithine decarboxylase . Putrescine formation may also occur by decarboxylation of arginine to form agmatine which is hydrolyzed to give putrescine and urea. Arginine is also involved in ornithine formation by action of the enzyme arginase. Activation of methionine by S-adenosylmethionine synthetase forms S-adenosylmethionine which is decarboxylated, after which the propylamine moiety of activated methionine may be transferred to putrescine to form spermidine or the polyamine moiety may be transferred to spermidine to form spermine. Hence, putrescine serves as a precursor to spermidine and spermine and additionally has been shown to have a marked regulatory effect upon the polyamine biosynthetic pathway in that it has been shown that increased synthesis of putrescine is the first indication that a tissue will undergo renewed growth processes. Cadaverine which is the decarboxylation product of lysine has been 7 0 81 shown to stimulate the activity of S-adenosylmethionine decarboxylase and is known to be essential to growth processes of many microorganisms, for example, H. parainfluenza.

The compounds of general Formula I wherein Z is R_HN(CH2) - and the lactams thereof are inhibitors of ornithine decarboxylase and lysine decarboxylase respectively as n varies from 3 to 4. The compdunds of general Formula 1 wherein Z is β-methylthioethyl or S-(5'-desoxy10 adenosin)5’-yl)^-methylthioethyl are inhibitors of Sadenosyimethionine decarboxylase and wherein Z is y-guanidinopropyl are inhibitors of arginine decarboxylase. As inhibitors of the above-enumerated decarboxylase enzymes the compounds of general Formulas I and 11 wherein Z is other than β-thioethyl, β-benzylthioethyl or Ya H00C-C-(CH2)2-S-S-(CH2)2- are useful as anti infective lilHe agents being effective in the control of microorganisms, for example, bacteria and viruses which are dependent upon polyamines for growth, for example, E. coli, Enterobacter, Klebsiella, Staphylococcus aureus, C. cadaver is, viruses such as, H. parainfluenza, picornaviruses, for example, encephalomyocarditis, Herpes simplex, poxviruses and arboviruses, for example Semiiki forest. The compounds of general Formulas I and II wherein Z is other than β-thio25 ethyl, β-benzylthioethyl, H00C-C-(CH2)2-S-S-(CH2)2- and l!lH2 R_HN(CH2)4- are also useful in the control of certain 3 ^7081 rapid growth processes. For example, the compounds are useful in the inhibition of spermatogenesis and embryogenesis and therefore the compounds find use as male antiferti1ity agents and abortifacients. The compounds are also useful in the inhibition of the immune response, thus the compounds are useful as immunosuppressants, and are useful in the control of neoplastic growth, for example, solid tumors, leukemias and lymphomas.

The compounds are also useful as inhibitors of prostatic hypertrophy, excessive scalp ceil growth as found with the occurrence of dandruff and as inhibitors of abnormal cutaneous cell growth as found with a psoriatic condition. The utility of compounds of general Formula I wherein Z is other than β-thioethyi, β-benzylthioethyl or Ya HOOC-C-(CH2)2-S-S-(CH2)2- as irreversible inhibitors of NH2 ornithine or S-adenosylmethionine decarboxylase enzymes in vivo can be demonstrated as follows. An aqueous solution of an appropriate compound of Formula I is given orally or parenterally to male mice or rats. The animals are sacrificed 1 to 48 hours after administration of the compound, and the ventral lobes of the prostate removed and homogenized with the activity of ornithine or S-adenosylmethionine decarboxylase enzyme being measured as generally described by E.A, Pegg and H.G. Wi1iiams-Ashman, Biochem. J. 108, 533-539 (1968) and J. Janne and H.G.

Wi11iams-Ashman, Biochem. and Biophys. Res. Comm. 42, 222-228 (1971). 7 0 81 The compounds of general Formula I wherein Ri is Υε hydroxy and I is other than HOOC-C-(CH2)2-S-S-(CH2)2I NH2 are useful as chemical intermediates for the preparation of novel cephalosporin derivatives which are useful as anti5 biotics and have the following general structure: Y 1 ς Z-C-CONH—i— L J—CH2X Formula III ο I COOM wherein Z and Y have the meanings defined in general Y2 Formula 1 except Z is not HOOC-C-(CH2)2-S-S-(CHa)2-; M is I NH2 hydrogen or a negative charge; and X is hydrogen or acetoxy.

The compounds of general Formula III and the pharmaceutically acceptable salts and individual optical isomers thereof are novel compounds useful as antibiotics and can be administered in a manner similar to that of many well known cephalosporin derivatives, for example, cephalexin, cephalothin, or cephaloglycine. The compounds of general Formula III and pharmaceutically acceptable salts and isomers thereof can be administered alone or in the form of pharmaceutical preparations either orally or parenterally and topically to warm blooded animals, that is, birds and mammals, for example, cats, dogs, bovine cows, sheep, horses and humans. For oral administration the compounds can be administered in the form of tablets, capsules or 4?ϋ8ί pills or in the form of elixirs or suspensions. For parenteral administration, the compounds may best be used in the form of a sterile aqueous solution which may contain other solutes, for example, enough saline or glucose to make the solution isotonic. For topical administration the compounds of general Formula III, salts and isomers thereof may be incorporated into creams or ointments.

Illustrative examples of bacteria against which the compounds of general Formula II! and the pharmaceutically acceptable salts and individual optical isomers thereof are active are Staphylococcus aureus, Salmonella schotmuehieri, Klebsiella pneumoniae, Dipiococcus pneumoniae and Streptococcus pyogenes.

Illustrative pharmaceutically acceptable non-toxic inorganic acid addition salts of the compounds of general Formula III are mineral acid addition salts, for example, hydrogen chloride, hydrogen bromide, sulfates, sulfamates, phosphate, and organic acid addition salts are, for example, maleate, acetate, citrate, oxalate, succinate, benzoate, tartrate; fumarate, malate and ascorbate. The salts can be formed by conventional means.

Illustrative examples of compounds of general Formula ill are 7-f[2,5-diamino-2-difluoromethylvaleryl]amino]3-acetyloxymethy1-8-oxo-5-thia-l-azab icyclo[ 4.2.0]oct-2ene-2-carboxylic acid, 7-[[2,6-diamino-2-fiuoromethylcaproyl]-j5-acetyloxymethyl-8-oxo-5-thia-l-azabicyc1o[4.2.0]oct-2-ene-2-carboxy]ic acid and 7-[[2-amino-2-trifluoromethyl - &-guan id inovaleryl ]amino]-J-acetyloxymethyl-8-oxo5-thia-l-azabicycio[4.2.0]oct-2-ene-2-carboxylic acid.

The preparation of the compounds of general Formula 111 is described hereinbelow.

As pharmacologically useful agents the compounds of general Formulas I and 11 can be administered in various manners to the patient being treated to achieve the desired effect. The compounds can be administered alone or in combination with one another. Also, the compounds can be administered in the form of a pharmaceutical preparation.

The compounds may be administered orally, parenterally, for example, intravenously, intraperitoneally, or subcutaneously, or topically. The amount of compound administered will vary over a wide range and can be any effective amount. Depending on the patient to be treated, the condition being treated and the mode of administration, the effective amount of compound administered will vary from about 0.1 mg/kg to 500 mg/kg of body weight of the patient per unit dose and preferably will be about 10 mg/kg to about 100 mg/kg of body weight of patient per unit dose.

For example, a typical unit dosage form may be a tablet containing from 10 to 300 mg of a compound of Formulas I or II which may be administered to the patient being treated 1 to 4 times daily to achieve the desired effect.

As used herein the term patient is taken to mean warm blooded animals such as mammals, for example, cats, dogs, rats, mice, guinea pigs, horses, bovine cows, sheep and humans.

The solid unit dosage forms can be of the conventional type. Thus, the solid form can be a capsule which can be of the ordinary gelatin type containing a novel compound of this invention and a carrier, for example, lubricant >♦708 j and inert fillers, such as lactose, sucrose and corn starch. In another embodiment, the novel compounds are tableted with conventional tablet bases such as lactose, sucrose or corn starch in combination with binders such as acacia, corn starch or gelatin, disintegrating agents such as corn starch, potato starch, or alginic acid, and a lubricant such as stearic acid, or magnesium stearate.

For parenteral administration the compounds may be administered as injectable dosages of a solution or suspension of the compound in a physiologically acceptable diluent with a pharmaceutical carrier which can be a sterile liquid such as water and oils with or without the addition of a surfactant and other pharmaceutically acceptable adjuvants. Illustrative of oils which can be employed in these preparations are those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, and mineral oil. In genera), water, saline, aqueous dextrose, and related sugar solutions, ethanols and glycols such as propylene glycol or polyethylene glycol are preferred liquid carriers, particularly for injectable solutions.

The compounds can be administered in the form of a depot injection or implant preparation which may be formulated in such a manner as to permit a sustained release of the active ingredient. The active ingredient can be compressed into pellets or small cylinders and implanted subcutaneously or intramuscularly as depot injections or implants. Implants may employ inert materials such as biodegradable polymers or synthetic silicones, for example, 7 0 81 Silastic, silicone rubber manufactured by the Dow-Corning Corporation.

The compounds of general Formula I wherein Z is βmethylthioethyl, β-thioethyl or RaHN(CH2)n-, Rx is hydroxy and each of R= and R, is hydrogen are prepared by treating respectively art ester derivative of methionine homocysteine ornithine or lysine wherein the amino group(s) is suitably protected and wherein the mercapto group of homocysteine is suitably protected with a strong base to form the carb10 anion intermediate which is reacted with a suitable halomethyl-halo alkylating reagent in an aprotic solvent, such as, dimethylsulfoxide, dimethylformamide, dimethylacetamide benzene, toluene, ethers, such as, tetrahydrofuran, diethyl ether or dioxane and in the presence of hexamethylphosphor15 triamide when Y is other than F2CH- at a temperature of about -12O°C to 120°C, preferably about 25° to 50°C for about 1/2 hour to 48 hours followed by acid or base hydroly sis as represented by the following reaction sequence. i-CH-COORa , Zi-C COORe 1 ι 1 ι N=C-RT 1 strong ι N=C-R71 base , | 1 R© compounds 1 __ Re __1 alkylat ing 7reagent Y Y I 1 .2-C-C0QRe 1 dilute aqueous Ζχ-C-COORs acid/hydrazine 1 nh2 £- N=C-R7 compounds 5 Re Hs0 (ac id/base) compounds 2 ψ Υ I Za-C-COOH I ΝΗ2 Formula IV In the above reaction sequence Re is a lower alkyl group, for example, methyl, ethyl, isopropyl, ri-propyl or n-butyl; R7 is hydrogen, phenyl, a straight or branched alkyl group having from 1 to 8 carbon atoms, methoxy or ethoxy; Ra is phenyl or a straight or branched alkyl group of .from 1 to 8 carbon atoms; or R7 and Rs taken together may form an alkylene group of from 5 to 7 carbon atoms, that is, -CH2-(CH2)m-CH2- wherein m is an integer of from to 5. Illustrative examples of straight or branched alkyl groups of from 1 to 8 carbon atoms which R7 and Re may represent are, for example, methyl, ethyl, n-propyl, isopropyl, ri-butyl, tert-butyl, £-pentyl, neopentyl or triethylmethyl; Zj is β-methylthioethyl; RgS(CH2)2wherein Rg is trityl, alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and is straight or branched, such as, methyl, ethyl, or isopropyl, alkoxycarbonyl wherein the alkoxy moiety has from 1 to 4 carbon atoms and is straight or branched, such as, methoxy, ethoxy, isopropoxy, phenylcarbonyl or benzyloxycarbonyl; Rio(CH2)n- wherein n is the integer 3 or 4 and Ri0 is -N=C-R7, wherein R7 and Ra have the meanings defined R« 0 0 I |l above, -NH-C-Rn or -NH-CORi2 wherein each of Rn and R12 is phenyl, benzyl or a lower alkyl group of from 1 to carbon atoms that is straight or branched, for example, methyl, ethyl or isopropyl; Z2 is β-methylthioethyi, 7081 RgS(CH2)2- or Ria(CH2)n- wherein Ria is -NH2, -NH-C-Rn II or -NH-CORi2 wherein Rg, Rn and Ria have the above defined meanings; Z3' is β-methylthioethyl, β-thioethyl or H2N(CH2)n- wherein n is the integer 3 or 4; and Y has the meaning defined in Formula I with the proviso that when Z2 is β-methylthioethyl or β-thioethyl Y is FCH2-, F2CH-, or F3C-. When in compounds 1 Ζχ is Rio(CH2)n- wherein Ri0 is -N=C-R7 each R7 and Rs is the same.

I Re Suitable strong bases which may be employed in the 10 above reaction sequence to form the carbanion intermediate are those which will abstract a proton from the carbon atom alpha to the carboxy group, such as, alkyl lithium, for example, butyl lithium or phenyl lithium, lithium di-alkylamide, for example, lithium diisopropylamide, lithium amide, tertiary potassium butylate, sodium amide, metal hydrides, for example, sodium hydride or potassium hydride, tertiary amines, such as, triethylamine, lithium acety]ide or dilithium acetylide. Lithium acetylide, di 1ithium acetylide, sodium hydride, and lithium diisopropylamide are particularly preferred bases.

Suitable alkylating reagents which may be employed in the above reaction sequence are illustratively chlorofluoromethane, bromofluoromethane, fluoro iodomethane, chlorodifluoromethane, bromodifIuoromethane, cfifluoro25 iodomethane, bromotrifiuoromethane, chlorotrifluoromethane, tri fluoro iodomethane, bromochloromethane, dichloromethane, chloroiodomethane, bromodichloromethane and dichloroiodomethane. The alkylating reagents are known in the art.

Removal of the protecting groups of the amine, mercapto and carboxylic function may be achieved in one step by treatment of compounds 2 with aqueous acid, for example, hydrochloric acid or toluene sulfonic acid at a temperature of about 0° to l6o°C for about 4 to 24 hours to give compounds of general Formula IV. It is preferred to remove first the protecting groups of the amine function(s) of compounds 2 when said functions are protected as a Schiff's base by treating compounds 2 with dilute aqueous acid, for example, hydrochloric acid or with hydrazine or phenylhydrazine in solvents, such as, lower alcohols, for example, methanol or ethanol, ethers, chlorinated hydrocarbons, benzene and water. Removal of the protecting groups of the carboxylic functions, the mercapto group and the amine group(s) when the amine group(s) is protected other than as a Schiff's base is achieved by treatment of compounds 3 with concentrated aqueous acids, for example, hydrobromic acid at a temperature of about 0° to l60°C or in aqueous bases, for example, ammonium hydroxide. Removal of the mercapto protecting group is carried out in the absence of oxygen, for example, under a nitrogen atmosphere.

The amine protected ester derivatives, that is, compounds 1 wherein Zi is β-methylthioethyl, RgS(CH2)2- or Rio(CH2)n- wherein n, R9 and Rio have the meanings defined above are prepared, when R7 is other than methoxy or ethoxy, by treating an appropriate amino acid ester with a carbonyl bearing compound to form a Schiff's base in a generally known manner, specifically, (a) when R7 is hydrogen, by treating the appropriate amino acid ester with benzaldehyde or an alkanal having from 1 to 9 carbon 1 atoms being straight or branched, for example, 1-propanal, 1-butanal, 2,2-dimethylpropan-l-al or 2,2-diethylbutan-lal; (b) when RT is phenyl by treating the appropriate amino acid ester with benzophenone or phenyl alkyl ketone wherein the alkyl moiety has from 1 to 8 carbon atoms and is straight or branched, for example, phenyl methyl ketone, phenyl ethyl ketone, phenyl isopropyl ketone, phenyl £butyl ketone.or phenyl tert-butyi ketone; and (c) when R7 is a straight or branched alkyl, group having from 1 to 8 carbon atoms, treating the appropriate amino acid ester with a phenyl alkyl ketone as described above or with a di-alkyl ketone wherein each alkyl moiety has from 1 to 8 carbon atoms and is straight dr branched, for example, dimethyl ketone, diethyl ketone, methyl isopropyl ketone, di-ji-butyl ketone or methyl tert-butyl ketone. The carbonyl bearing compounds are known in the art or may be prepared by procedures well known in the art.

When RT is methoxy or ethoxy, an appropriate amino acid ester derivative is reacted with benzoyl halide, for example, chloride or an alkanoic acid halide, for example, chloride wherein the alkanoic acid has from 1 to 9 carbon atoms and may be straight or branched, such as, acetyl chloride, propionyl chloride, butyryl chloride, tertbutyryl chloride, 2,2-diethylbutyric acid chloride or valeryl chloride, at 0°C In ethers, methylenechloride, dimethylformamide, dimethylacetamide or chlorobenzene in the presence of an organic base such as triethylamine or pyridine after which the reaction mixture is allowed to warm to about 25°C for one hour. The resulting amide derivative is combined with an alkylating reagent, such as, methylfluorosuifonate, dimethylsulfate, methyliodide, methyl p-toluenesulfonate or trimethyloxonium hexafluorophosphate when R? is methoxy or triethyloxonium tetrafluoroborate when R7 is ethoxy at about 25°C in a chlorinated hydrocarbon solvent such as methylene chloride, chlorobenzene or chloroform, and the reaction mixture is refluxed for about 12 to 20 hours. The mixture is then cooled to about 25°C and an organic base such as triethylamine or pyridine is added after which the solution is extracted with brine and the product isolated.

When in compounds 1 R7 and Re together form an alkylene group of from 5 to 7 carbon atoms said amino acid ester derivatives are obtained by treating the amino acid ester with a cyclic alkanone selected from cyclopentanone, cyclohexanone and cycloheptanone to form a Schiff's base by procedures generally known in the art.

When Zi is RsS(CH2)2- in compounds 1 the mercapto protecting group is added to homocysteine in the absence of oxygen for example, under a nitrogen atmosphere by procedures generally known in the art, for example, L. Zervas and I. Photaki, J. Am. Chem. Soc. 84, 3θθ7 (1962), L, Zervas, et ai.,J. Am, Chem. Soc. 85, 1357 (1963) and A. Berger, et al., J. Am. Chem. Soc. 73, 4483 (1956). Generally, when Ra is trityl a slight excess of trityl chloride is added to homocysteine in a solvent such as dimethylformamide. When Rs is alkylcarbonyl or phenylcarbonyl slightly less than 1 equivalent of an appropriate acid halide, for example, acetyl chloride, propionyl chloride or benzoyl chloride in an ether solvent, such as, tetrahydrofuran is added to 1 equivalent of homocysteine 7 0 81 in an aqueous base, such as, aqueous sodium carbonate.

Similarly when Rg is alkoxycarbonyi or benzyloxycarbonyl slightly less than 1 equivalent of an appropriate alkyl haloformate such as methyl chloroformate or ethyl chloro5 formate or benzyl chloroformate in an ether’soivent is added to 1 equivalent of homocysteine in aqueous base.

II When Zi is Ri0(CH2)n- wherein Ri0 is -NH-CRu or 0 0 I II II · -NH-CORi2 the -CRu and -CORi2 protecting groups are added to the corresponding free amino acids, that is, ornithine and lysine, by treatment of said amino acid with an excess of copper salt, for example, copper carbonate in boiling water for about 1 to 6 hours, and upon cooling to room temperature the insoluble materials are filtered off, and the filtrate is treated with an appro0 ll priate acid halide when Ri0 is -NH-CRu or an appropriate II alkyl or aryl haloformate when Ri0 is -NH-CORi2, for example, in acetone in the presence of a base such as sodium bicarbonate or sodium hydroxide followed by treatment with hydrogen sulfide. Illustrative acid halides which may be employed are acetyl chloride, propionyl chloride, benzoyl chloride or 2-phenylacetyl chloride. Illustrative haloformates which may be employed are benzyl chloroformate, phenyl chloroformate, methyl chloroformate or ethyl chloroformate.

The amino acid ester is formed by generally known procedures, for example, the amino acid is treated with an appropriate alcohol, such as, methanol, ethanol, or nbutanol saturated with HCl gas.

The compounds of general Formula I wherein i is γguanidinopropyl are prepared from the corresponding suitably protected derivative wherein Z is R,HN(CH2) - wherein ο Π Rg is hydrogen and n is the integer 3 and wherein Y is FCH2-, F2CH-, F3C-, that is, the compound Y' H2N(CH2)3-C-COOH Formula V NHRb wherein Y' is FCH2-, F2CH-, or F3C- wherein R^ has the meaning defined in Formula I by treatment with an alkylisothiouronium salt, for example, ethylisothiouronium hydrobromide by procedures generally known in the art, for example, Organic Synthesis III, p. 440 (1955). The reaction is carried out in the presence of a base, for example, aqueous sodium.hydroxide or potassium hydroxide at a pH of about 8-12 and at a temperature of about 0° to 100°C for about 6 hours to 8 days after which the reaction mixture is neutralized with concentrated mineral acid, for example, hydrochloric acid and the product isolated. When R^ is hydrogen the α-amino group may be protected with, for example, a benzyloxycarbonyl group. When R^ is II -C-CH-R2 the free amino group is protected prior to the I nh2 reaction with, for example, benzyloxycarbonyl. The amine protecting group is subsequently removed by acid hydrolysis, for example, with hydrochloric acid.

The compounds of general Formula I wherein Z is S-(5’~ desoxyadenosin-5'-yl)-S-methylthioethyl are prepared by reacting 5'-p-toluenesulfonyl adenosine, 5'-chloroadenosine or 5'-bromoadenosine having the structure optionally protected as the 2',3'-isopropyIidene NH, CH2-Ri4 Formula VI wherein Ri4 is p-toluenesulfonyl, chlorine or bromine with a metallic salt of an appropriate a-halomethylhomocysteine of the structure NH2 Formula VII wherein Y1 is FCH2-, F2CH- or F3C- and M+ is sodium or lithium in liquid ammonia for about 1 to 5 hours to formthe S-adenosyl-α-halomethylhomocysteine derivative which is hydrolyzed with acid, such as, hydrochloric acid when the compound of Formula Vi is protected as the 2',3'-isopropyl idene then treated with methyl iodide in acidic solvents, such as, formic acid, acetic acid or mixtures thereof.

The metallic salts of the halomethylhomocysteine derivative, that is, the compounds of Formula VII are obtained by the addition of sodium or lithium metal to an appropriate α-halomethylhomocysteine, a-haiomethyihomocystine or α-halomethyi-S-benzylhomocysteine wherein in '17 0 81 each compound type the halomethyl group is FCH2-, FeCH- or FgC- in liquid ammonia until the blue color persists for 15 minutes.

The a-halomethyl-S-benzylhomocysteine derivatives are prepared by treating the corresponding α-halomethylmethionine, the synthesis of which is described hereinabove, with benzyl chloride by procedures generally known in the art, for example, Biochemical Preparations 5, 91 (I957). The reaction is carried out in concentrated hydrochloric acid at reflux temperature for about 24 hours after which the reaction mixture is concentrated under reduced pressure, neutralized with base, for example, ammonia and the product isolated. The a-halomethyl-Shomocystine derivatives are prepared by air oxidation of an aqueous solution of the corresponding halomethy,homocysteine at a pH of about 4 to 8 in the presence of a catalytic amount of a ferric salt, for example, ferric chlor ide.

Following is described the preparation of compounds of general Formula I wherein Rg and/or R^ are other than hydrogen including compounds of general Formula V. The following description is applicable to all the above said compounds, however, it is necessary to protect one or the other of the amino groups prior to treatment with the appropriate reactant, that is, acid halide or anhydride, alkyl haloformate or acid of the formula HOOCCH-R2 or NH2 anhydride thereof as described below to give compounds wherein either or both of Rg and Rb is other than hydrogen as follows: When Rg is hydrogen and R^ is other than hydrogen, the amino group to which Re is attached is protected as a copper salt by treatment of the corresponding derivative wherein Rg and R^ are hydrogen with an excess of a copper salt, for example, copper carbonate after which the amino group to which R is attached is protected with, a for example, benzyloxycarbonyl or tert-butoxycarbonyl by treatment with benzyl chloroformate or tert-butoxycarbonyi azide respectively followed by treatment with hydrogen sulfide, by procedures generally known in the art and illustrated more fully in the specific examples contained herein, prior to treatment with the appropriate reactant described below to give compounds wherein R^ is other than hydrogen. The Rg amine protecting group is subsequently removed by treatment with acid, for example, trifluoroacetic acid, HBr in dioxane or HBr in acetic acid or hydrogenolysis. The thus obtained compounds, that is, compounds wherein R is hydrogen and R, is other than hydrogen may be treated with the appropriate reactants described below to give compounds wherein Rg and R^ are both other than hydrogen and may be the same or different or to give compounds of Formula I wherein Z is γ-guanidinopropyl as described hereinabove. In preparing compounds wherein Rg is other than hydrogen and R^ is hydrogen the amino group to which R^ is attached is protected as a copper salt by treatment of the corresponding derivative wherein each Rg and R^ is hydrogen with an excess of copper salt, for example, copper carbonate prior to treatment with the appropriate reactant described below followed by acid or base hydrolysis and subsequently treating with hydrogen sulfide. 4^08] The compounds of general Formulas I and V wherein R, or Rb is alkylcarbonyl wherein the alkyl moiety is straight or branched and has from 1 to 4 carbon atoms are prepared by treating the corresponding derivatives wherein Rg or R^ is hydrogen or is suitably protected or as to compounds of Formula I, Rb is other than hydrogen as described above and Ri is hydroxy with an acid halide of the formuta • ! RisC-haio wherein halo is a halogen atom, for example, chlorine or bromine and Ris is a straight or branched alkyl group having from 1 to 4 carbon atoms or an appropriate acid anhydride, in water in the presence of a base such as sodium hydroxide or sodium borate at a temperature of about 0° to 25°C for about 1/2 hour to 6 hours. When appropriate, protecting groups are removed as described hereinabove by treatment with acid or hydrogenolysis.

The compounds of general Formulas I and V wherein Rg or Rg is alkoxycarbonyi wherein the alkoxy moiety is straight or branched and has from 1 to 4 carbon atoms and Ri is hydroxy are prepared by treating the corresponding derivative wherein Rg and R^ is hydrogen or is suitably protected or as to compounds of Formula 1, R& is other than hydrogen as described hereinabove with an alkyl halo0 formate of the formula halo-C-ORie wherein halo is a halogen atom such as chlorine or bromine and Rie is a straight or branched alkyl group having from 1 to 4 carbon atoms in water in the presence of a base such as sodium hydroxide or sodium borate at a temperature of about 0° to 25°C for about 1/2 hour to 6 hou-s. When appropriate, protecting 4-7081 groups are removed as described hereinabove by treatment with acid or hydrogenolysis.

The compounds of general Formulas I and V wherein 0 RQ or is -C-CH-R2 wherein Re is hydrogen, a straight or NHS branched iower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzyl and Rt is hydroxy are prepared by treating the corresponding derivative wherein R. or R^ is hydrogen or is suitably protected or as to compounds of Formula I, R^ is other than hydrogen as described here10 inabove with an acid of the formula HOOC-CH-R2, or an NH2 anhydride thereof, wherein the amino group is protected with a suitable blocking group such as benzyloxycarbonyl or tert-butoxycarbonyl and R2 has the meaning defined hereinabove in an ether, such as, tetrahydrofuran or dioxane, methylene chloride or chloroform and in the presence of a dehydrating agent, such as, dicyclohexylcarbodiimide when the free acid is employed, at a temperature of about 0° to 35°C for about 1 to 12 hours followed by acid and base hydrolysis and when appropriate, hydrogenolysis to remove the protecting groups.

The compounds of the general Formula I wherein Rx is a straight or branched alkoxy group of from 1 to 8 carbon atoms are prepared by converting the corresponding compound wher.ein Rx is hydroxy to the acid halide by, for example, treatment with thionyl chloride, followed by alcoholysis with an alcohol of the formula Ri70H wherein Rl7 is a straight or branched alkyl group having from 1 to 4-7t.JSl carbon atoms by procedures generally known in the art. Alternatively, compounds of general Formula I wherein Rx is a straight or branched alkoxy group of from 1 to 8 carbon atoms may be prepared from the corresponding deriva5 tive wherein Ri is hydroxy by treatment of said derivative with an alcohol of the formula Ri70H as defined above saturated with HCl for about 3θ minutes for 12 hours at .a temperature of about 25°C to the boiling point of the alcohol. 7081 The compounds of this invention wherein Ri is -NR4R5 wherein each of R4 and Rs is hydrogen or a lower alkyl group of from 1 to 4 carbon atoms are prepared by an acylation reaction of an acid halide, for example, an acid chloride, of the corresponding compound wherein Ri is hydroxy and Rg and R^ have the meanings defined in general Formula I with the proviso that any free amino group is suitably protected with groups, such as, carbobenzyloxy or tert-butoxycarbony I with an excess of an appropriate amine which may be represented as HNR4R5. The reaction is carried out in methylene chloride, chloroform, dimethyl formamide, or ethers such as tetrahydrofuran and dioxane, or benzene at about 25°C for about 1 to 4 hours. Suitable amines are ammonia, or a compound which is a potential source of ammonia, for example, hexamethylenetetramine; primary amines, for example, methylamine,|ethylamine or n-propylamine; and secondary amines, For example, dimethylamine, diethylamine, or di-£-butylamine. Following the acylation reaction the protecting groups are removed by treatment with acid, for example, trifluoroacetic acid or hydrogen bromide in dioxane.

The compounds of general Formula I wherein Ri is -NH-CH-COOH are prepared by reacting the corresponding Rs derivative wherein Ri is hydroxy or a functional derivative thereof, such as, an acid anhydride and Rg and R^ have the meanings defined in Formula ! with the proviso that any free amino group is protected with a suitable blocking group, such as, benzyloxycarbonyi, tert-butoxycarbony1 by 7 0 81 reacting the amine protected free acid with a compound of the structure NH2-CH-COORia wherein R3 has the meaning Rs defined in general Formula 1 and R18 is a lower alkyi group, for example, methyl or ethyl in an ether solution, such as, tetrahydrofuran or dioxane at about 0°C to 35°C for about 1 to 20 hours followed by acid then base hydrolysis, for example, with 2 N aqueous NH3 at about 0° to 50°C for about 1 to 20 hours, to remove the protecting group(s), with the proviso that when the amine protected free acid is employed the reaction is carried out using a dehydrating agent such as dicyclohexylcarbodiimide.

The lactams of the compounds of general Formula I wherein each of Rg and R^ is hydrogen and Ri is hydroxy are prepared from the corresponding amino acid ester of the structure Y HSN(CHZ) 4-C0Ri3 Formula VIII I nh2 wherein n and Y have the meanings defined in Formula I, and Rig is a straight or branched alkoxy group of from 1 to 8 carbon atoms, i1lustrativeiy methoxy, ethoxy, isopropoxy, butoxy, or hexyloxy; and more preferably when Y is ClCH2from the corresponding ct-hydroxymethyl substituted amino acid ester of the structure CH20H I H2N(CH2)n-C-C0Rls Formula IX NH2 wherein n and Rig have the meanings defined in Formula VIII by treating said amino acid esters with an appropriate base 7 0 81 such as, sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, potassium carbonate, sodium methoxide, potassium methoxide, potassium tert-butoxide, sodium amide, or an organic amine such as a trialkylamine, for example, triethylamine in a solvent such as a lower alcohol, for example, methanol, ethanol, isopropyl alcohol, .n-butanol, water, dimethylformamide, dimethylsulfoxide, hexamethylphosphortriamide or mixtures of these solvents for from 1/2 hour to 24 hours at a temperature of from about 0° to 12O°C optionally under a nitrogen atmosphere, with the additional step when Y is C1CH2- of treatment with a chlorinating agent, such as, thionyl chloride, phosphorus oxychloride or phosphorus pentachloride in a solvent such as formamide dimethylformamide or dimethyl15 acetamide for about 12 to 36 hours at a temperature of about 40° to 120°C.

The compounds of general Formula VIII are obtained by procedures generally known in the art from the corresponding amino acid, for example, by treating said amino acid with an appropriate alcohol, for example, methanol, ethanol, isopropyl alcohol, n-butanol or ri-heptanol saturated with HCl gas.

The compounds of general Formula IX are obtained by the general method described in Synthesis 1973» 792, for example, by treating 1 equivalent of ornithine or lysine with 2 equivalents of benzoyl chloride then 2 equivalents of a base, such as, sodium hydroxide to form the bisamide which is treated with an acid anhydride, for example, acetic anhydride at about 90°C for about 1/2 hour followed by treatment with aqueous formaldehyde and pyridine at 7 0 8 1 about 25°C for about 8 to 24 hours then treatment with water to give the oxodioxane which is treated with a catalytic amount of sodium methoxide in methanol then neutralized and treated with acid, for example, hydrochloric acid at about 120°C for about 2 to 24 hours.

The individual optical isomers of the compounds of general Formula I wherein Z is RgHN(CH2)n- wherein each of R^ and is hydrogen and Ri is hydroxy are obtained from the lactam of said compounds using a (+) or (-) binaphthylphosphoric acid salt by the method of R. Viterbo et al., Tetrahedron Letters 43, 4617 (1971)· Other resolving agents such as (+) camphor-10-su1fonic acid may also be employed. Similarly the individual optical isomers of compounds of Formula 1 wherein Z is β-methylthioethyl, R is hydrogen and Rt is hydroxy, are obtained from the free amino acid using a (+) or (-) binaphthylphosphoric acid salt or other agents such as (+) camphor-10-su1fonic acid.

The individual optical isomers of compounds of general Formula 1 wherein Z is γ-guanidinopropyl are obtained as described herein for the racemate only starting with the resolved ornithine analog.

The individual optical isomers of compounds of Formula I wherein Z is β-benzyIthioethyl may be obtained as described herein for the racemate only starting with the resolved analog wherein Z is β-methylthioethyl. The individual optical isomers of compounds of Formula I wherein Z is β-thioethyl may be prepared from the resolved derivative wherein Z is β-benzy1thioethyl, and the individual optical isomers of compounds of Formula I wherein Ya H2N Z is H00C-C-(CH2)a-S-S-(CH2)2- may be prepared from the NH2 resolved derivative wherein Z is β-thioethyl by procedures generally known in the art. The individual optical isomers of compounds of general Formula I wherein R is other than hydrogen and Ri is other than hydroxy are obtained as described herein for the racemate only starting with the resolved free amino acid.

The compounds of general Formula 111 are prepared by coupling a 7-aminocephalosporanic acid or a derivative thereof having the formula ’CH2X Formula IX COOM .wherein M is hydrogen or a negative charge and X is hydrogen or acetoxy, with an acid of the formula Y I Z-C-COOH Formula X I NH2 or a functional derivative thereof, such as, the acid chloride or an acid anhydride in the presence of a dehydrating agent such as dicyclohexylcarbodiimide when the free acid is employed wherein Z and Y have the meaning defined in general Formula I except Z is not Y2 HOOC-C-(CH2)2-S-S-(CH2)2- and the amino group is proI NH2 tected with a suitable blocking group, for example, tertbutoxycarbonyl followed by acid hydrolysis to remove the 7081 amino protecting groups.

The coupling reaction is generally carried out in a solvent, such as, ethyl acetate, dioxane, chloroform or tetrahydrofuran in the presence of a base, such as, alkaline bicarbonate. The temperature of the reaction may vary from about -10° to 100°C, and the reaction time may vary from about 1/2 hour to 10 hours. The cephalosporin products are isolated by conventional procedures.

The compounds of Formula X are prepared by procedures described hereinabove, and the compounds of Formula IX are commercially available.

The following Example 1 illustrates the use of a compound of general Formula I wherein Ri is hydroxy as a chemical intermediate in the preparation of a cephalosporin of Formula III.

EXAMPLE 1 7- ( f 2,5-Diamino-2-d i fluoromethvIvalery 1]amino]-3-acetyloxymetny 1-d-oxo-3- th ia-l-azab i cycl of 4-.2.0 ;oct-2-ene-2-~ carboxy lie acid A mixture of 1 g of 3-acetyloxy-7-amino-8-oxo-5-thial-azab icyc lo[ 4.2. 0]oct-2-ene-2-carboxy 1 ic acid and 1 g of 2,5-diamino-2-difuloromethyiva1eric acid chloride wherein the free amino groups are protected with tert-butoxycarbonyl in 5θ ml of ethylacetate is refluxed for 2 hours after which the solvent is removed leaving a residue which is treated with mild acid and chromatographed on silica gel using benzene-acetone as the eluant to give 7-((2,5diamino-2-d i fluoromethyl valeryl]amino]-3-acetyloxymethyl8- oxo-5-thia-l-azabieye Io[4.2.0]oct-2-ene-2-carboxylic acid.

EXAMPLE 2 An illustrative composition for hard gelatin capsules is as follows: (a) a, δ-diamino-a-difluoromethyI- 20 mg valeric acid (b) talc 5 mg (c) lactose 9θ mg The formulation is prepared by passing the dry powders of (a) and (b) through a fine mesh screen and mixing them well. The powder is then filled into hard gelatin capsules at a net fill of 115 mg per capsule.

EXAMPLE 3 An i1lustratfve composition for tablets is as follows (a) a-amino-a-fluoromethyl-6- 20 mg guanidinovaleric acid (b) starch 43 mg (c) lactose 45 mg (d) magnesium stearate 2 mg The granulation obtained upon mixing the lactose with the compound (a) and part of the starch and granulated with starch paste is dried, screened, and mixed with the magnesium stearate. The mixture is compressed into tablets weighing 110 mg each.

EXAMPLE 4 An illustrative composition for an injectab.le suspension is the following 1 ml ampul for an intramuscular injection.

Weight per cent (a) α-amino-a-difluoromethyl-γ- 1.0 methylthiobutyric acid (b) polyvinylpyrrolidone 0.5 (c) lecithin 0.25 (d) water for injection to make 100.0 The materials (a)-(d) are mixed, homogenized, and filled into 1 ml ampuls which are sealed and autoclaved minutes at 121°C. Each ampul contains 10 mg per ml of novel compound (a).

The following examples further illustrate the compounds of the invention.

EXAMPLE 5 g-Amino-g-d i fluoromethyl-y-fS-(5'-desoxyadenos in-5'-yl)thiojbutyric acid To a solution of the disodium salt of g-difluoromethyl-homocysteine (2 mmole) [prepared in situ from adifluoromethyIhomocystine (1 mmole) a-difluoromethylmethionine (2 mmole) or a-difluoromethyl-S-benzylmethio15 nine (2 mmole) and sodium (2.1 mmole)] in liquid ammonia (100 ml) is added 5'-O-tosyladenosine (2 mmole). The reaction mixture is stirred for 2 hours and then the solvent is allowed to evaporate under a stream of nitrogen. The residue is dissolved in water ( 50 ml) and purified by passing first through an ion exchange resin column (KV-2 NH4+ form) and then a DEAE cellulose (OH form) column. The aqueous eluate is evaporated under reduced pressure and crystallization of the residue from water ethanol yields pure g-amino-g-difluoromethyl-y25 [S-(5'-desoxyadenosin)5'-yl)thio]butyric acid.

Similarly by using g-fluoromethyimethionine instead of α-difluoromethylinethionine, there is obtained g-aminoα-fluoromethyJ-y-[S-(5'-desoxyadenos in-5' - yl)thio]butyric acid.

EXAMPLE 6 g-Amino-g-d i fluoromethyi-y-f S-(5'-desoxyadenos in-51-ylΙέ-(methyl )thio]butyr ic acid To a solution of S-adenosyl-a-difluoromethylhomocysteine (600 mg) in a mixture of acetic acid (4 ml) and 5 formic acid (4 ml) is added methyl iodide (1 mi). The reaction mixture is kept under nitrogen in the dark for 8 days at room temperature. The solvent is removed under reduced pressure at room temperature and res idue taken up in 0.1 N hydrochloric acid (8 ml). A saturated solution of Reinecke salt is added until the precipitation is complete. The precipitated salt (about 1 g) is treated with silver sulfate (1.5 g) in dry acetone at room temperature for 36 hours. The insoluble residue is filtered off and washed with methanol. The combined filtrates are concentrated under reduced pressure to yield a-amino-gdifluoromethyl-y-[S-(5'-desoxyadenos in-5'-yi)-S-(methy1)thiojbutyric acid.

Similarly by using S-adenosyl-g-fiuoromethylhomocysteine instead of S-adenosyl-g-difluoromethylhomocys20 teine, there is obtained methyI-y-[S-(5'-desoxyadenosin5'-yl)-S-(methyl)thio]butyr ic acid.

EXAMPLE 7 3-Amino-3-difluoromethyl-2-pi per idone To a solution of methyl-2-difluoromethyl-2,5-diamino25 pentanoate-dihydrochloride (2.7 g) in dry methanol (30 ml) is added under nitrogen 2 equivalents of sodium methylate in methanol (0.46 g of sodium in 20 ml of methanol). The reaction mixture is stirred for 3 hours at room temperature then the solvent is evaporated under reduced pressure The residue is extracted with ether to yield crude 3-amino3-difluoromethyl-2-piperidone which is purified either by crystallization from CHCU/pentane: (m.p. : 149°C) or by distillation (b.p. 135°C/0.05 mmHg).

EXAMPLE 8 (-) and (+) 2-Amino-3-difluoromethyl-2-piperidone hydrochloride To a solution of (-) binaphthylphosphoric acid (BNPA) (1.27 g) in h°t ethanol (5θ ml) is added a solution of (+) 3-amino-3-difluoromethyl-2-piperidone (0.546 mg) in hot ethanol (5 ml). On cooling, crystals separate. The reaction mixture is then let stand at 4°C overnight. The precipitate is filtered off, washed with ethanol and diethyl ether to give 0.54 g of (-) binaphthylphosphoric salt (ta]D = -409°C = 0.3, MeOH mp: 300°C). Recrystallization of the mother liquor yields 0.15 g of (-) binaphthylphosphoric salt. Concentration of the filtrate gives 1.1 g of a sticky material which is treated with HCl 3 M at room temperature for 3 hours. The (-) BNPA is filtered off and the filtrate concentrated under reduced pressure. Recrystallization of the residue (320 mg) in ethanol affords (+) 3-θηιino-3-difluoromethyl-2-piperidonemonohydrochloride (l6o mg) ([α]θ = + 18°6, C = 1, MeOH) m.p. 238°C). Treated in the same condition the (-) BNPA salt (436 mg) gives (-) 3-amino-3-difluoromethyl-2-piperidone monohydrochloride (137 mg) which is recrystallized in ethanol (67 mg) ([alp = -19°, C = 1.02, MeOH; mp = 240°C dec.). (-) and (+) 2-difluoromethyl-2,5-diamino pentanoic acid monohydrochlor ide (-) 3*Difluoromethyl-3-amino-2-piperidone hydro4 7 0 81 chloride (6θ mg) is heated in HCl 6m (4 ml) at reflux for 12 hours. After concentration under reduced pressure, the residue is dissolved in water and the pH of the solution is adjusted to 4.5 with a solution of NEt3. The solution is then concentrated under reduced pressure and the residue extracted many times with chloroform and then recrystallized from H2O/EtOH to give (+) 2-difluoromethyl-2,5diamino pentanoic acid monohydrochloride (54 mg) ([α]θ= +6°, C =0,48; MeOH; mp> 240°C). By an identical treatment, (+) 5-difluoromethyl-3-amino~2-piperidone hydrochloride (96 mg) gives (-) 2-difiuoromethyi-2,5-diaminopentanoic acid monohydrochloride (56 mg) ([a]Q = -10°, C = 0.7 MeOH, mp > 244°).

EXAMPLE 9 2-Difiuoromethyi-2-amino-5-guan idinopentanoic acid To a solution of 2-difluoromethyi-2,5-diamino-pentanoic acid monohydrate’monohydrochioride (5 g or 21.13 mmole) in NaOH 2M (8.5 mi) is added at once ethylthiouronium hydrobromide (7*82 g or 42.26 mmole). The pH of the solution is adjusted to 10-5 with NaOH 2M and maintained to that value for 4 days. The reaction mixture is then neutralized to pH = 7 with 1M HCl and concentrated under reduced pressure. The residue is passed on an Amberlite IR 120 H+ form column. Elution with NH4OH 2M affords 2-difluoromethyl-2-amino-5-guanidinopentanoic acid which is recrystallized from H2O/EtOH (2.3 g) m.p. 257°C.

EXAMPLE 10 Methyl-2-di fluoromethyl-2,6-dibenzaId imine hexanoate To a solution of lithium diisopropyIamide (60 mmole prepared in situ in tetrahydrofuran, cooled to -78°C, magnetically stirred and kept under nitrogen, is added slowly a solution of methyl-2,6-dibenzaId imine hexanoate (16.82 g or 50 mmole) in dry THF (60 mi). The reaction mixture is allowed to rise to room temperature over a period of 2 hours and is then rapidly warmed up to 40°C.

The nitrogen inlet is replaced by a balloon filled with C1CHF2 (1 1). The reaction mixture is kept overnight at room temperature under stirring and then is hydrolyzed with water. Usual ether extraction yields methyl-2-difluoromethyl-2,6-dibenzylideneamino hexanoate.

EXAMPLE 11 Methyl-2-di fluoromethyl-2,6-d iaminohexanoate d ihydrochlor i de A solution of methyl-2-difluoromethyl-2,6-dibenzaldimine hexanoate (15-4 g or 40 mmole) in ether (20 ml) is magnetically stirred with 2N hydrochloric acid (110 ml) for 1 hour. Usual ether extraction and evaporation to dryness of the aqueous phase affords methyl-2-difluoromethyl-2,6-diamino hexanoate dihydrochloride (9·15 g 85$) which is crystallized from methanol, ether, m.p. 207°C.

EXAMPLE 12 2-Dif1uoromethyl-2,6-diaminohexanoic acid hydrochloride A solution of methyl-2-difluoromethyI-2,6-diamino hexanoate dihydrochloride (4 g, 14 mmole) in concentrated HCl is treated under reflux for 20 hours. After evaporation to dryness and decoloration with charcoal, the pale solid obtained is dissolved in absolute ethanol. The pH of the solution is adjusted to 4.5 with a solution of triethylamine (M) in absolute ethanol upon which the adifluoromethyl-lysine monohydrochloride precipitated. Recrystallization from water/ethanol yields analytically pure α-difluoromethyl lysine hydrochloride as white crystals, m.p. >260°C (dec.).

EXAMPLE 13 Methyl 2-chloromethyl-2,6-diaminohexanoate dihydrochloride A suspension of 2,6-diaminohexanoic acid hydrochloride (18.26 g 100 mmole) in dry methanol (100 ml) is saturated with gaseous hydrochloric acid and heated under reflux for 2 hours. Upon cooling, the lysine methyl ester dihydrochloride crystallizes in the reaction mixture. To an ice cooled suspension of methyl 2,6-diaminohexanoate dihydro15 chloride (4.66 g, 20 mmole) in methylene chloride (50 ml) is added benzaldehyde (4.04, 40 mmole) and then dropwise a solution of triethylamine (4.24 g, 40 mmole) in methylene chloride (30 ml). The reaction mixture is allowed to stand overnight at room temperature under stirring and then concentrated under reduced pressure. Usual ether work-up yields methyl-2,6-dibenzylidene amino hexanoate as an oily residue.

To a solution of lithium diisopropylamide (10 mmole; prepared in situ from a solution of IN di isopropylamine in tetrahydrofuran and a solution of butyl lithium 2N in hexane) in tetrahydrofuran and hexamethylphosphorustriamide (2.5 ml), cooled to -78°C, magnetically stirred and kept under nitrogen, is added slowly a solution of methyl-2,6dibenzylidene ami no-hexanoate (3.365 g, 10 mmole) in dry tetrahydrofuran (20 ml). The reaction mixture is kept under stirring at -78°C for 1/2 hour and then a solution of chlorobromomethane (1.3 g, about 10 mmole) in dry tetrahydrofuran (10 ml) is added rapidly. The temperature is allowed to rise slowly to room temperature and the stirring is continued overnight. The reaction mixture is quenched with water. Usual work-up gives methy1-2-chloromethyl-2,6dibenzylideneamino-hexanoate as an oily residue.

To a solution of methy!-2-chloromethy1-2,6-dibenzylidene amino hexanoate (2.85 g, ,7-4 mmole) in ether (6 ml), IN hydrochloric acid (20 ml) is added under vigorous stirring. The reaction mixture is stirred overnight at room temperature and then extracted in the usual way. The aqueous phase evaporated to dryness under vacuo yields methyl-2-chloromethyI-2,6-diamino hexanoate dihydrochloride as an oily and hygroscopic residue (2.05 g, 90^) [nmr (D20): 4.05 (2H, q)-CH2Cl; 3-87 (3H, s)-C02Me; 3.03 (2H, broad t, J = 7H.,) H2N-CH2-].

EXAMPLE 14 2-Chloromethyl-2,6-diaminohexanoic acid monochlorohydrate A solution of methyl-2-chloromethyl-2,6-diamino hexanoate dihydroehloride (2.05 g, 7·3 mmole) in concentrated hydrochloric acid (10 ml) is heated under reflux for 18 hours. Concentration under vacuo followed by treatment with charcoal affords an oily residue (2.1 g) which is dissolved in dry methanol. Upon addition of propylene oxide until pH 3-4, 2-chioromethyl-2,5-diaminohexanoic acid monochlorhydrate precipitates in the reaction mixture (1.25 9 or 75i®)· Recrystai1ization from water/ ethanol affords analytically pure 2-chloromethyl-2,6diaminohexanoic acid monochlorhydrate as white crystals, iTSOSl m.p. >216 (dec.).

EXAMPLE 15 2-Pifluoromethyl-2,5-diaminopentanoic acid Under nitrogen a solution (500 ml) of 2M butyi5 lithium in hexane is added to a stirred solution of 142.1 ml of di isopropylamine in 1.5 liters of tetrahydrofuran at -78°C after which 261 g (0.81 mole) of ornithine dibenzaldimine methyl ester in 1-5 liters of tetrahydrofuran is added. Upon completion of the addition the reaction temperature is raised to 4o°C and maintained between 40° and 50qC for 2 hours during which time chlorodifluoromethane gas is bubbled through the mixture with stirring. The reaction mixture is then treated with a saturated solution of sodium chloride. The organic material is extracted with ether, and the ether extract washed several times with sodium chloride solution, dried over magnesium sulfate and evaporated to give a viscous oil. The oil is stirred with INHCl (1.5 1) for 3 hours, the mixture extracted several times with chloroform and the aqueous solution evaporated to dryness. The oily residue is refluxed with 12N hydrochloric acid (1.5 I) for 16 hours, the cooled solution clarified by chloroform.extraction before concentration, decolorization (charcoal), and further concentration to about 75θ ml. The pH of the solution is adjusted to 2-5 by the addition of triethylamine, the solution treated again with charcoal before concentration to about 500 ml and dilution with 7-8 liters of acetone.

The precipitated product is filtered off and washed with ethanol. The crude product is recrystallized by dissolving in about 150 ml hot water and treatment of the solution ί 7 OS I with hot ethanol (450 ml). On cooling crystals of 2difJuoromethyl-2,5-diaminopentanoic acid hydrochloride monohydrate separate 71 g (37$), m.p. 183°C.

EXAMPLE 16 -Hydroxymethyl-3-amino-2-pi per idone 2- Hydroxymethyl-2,5-diaminopentanoic acid hydrochloride (5 g or 2.5 x 10 2 mole) is suspended in 75 ml of absolute methanol and the solution is saturated with dry hydrogen chloride. The homogenous solution is then heated under reflux for 48 hours. The reaction mixture is regularly saturated with dry hydrogen chloride. The solvent is evaporated under reduced pressure and the hygroscopic residue is dried under high vacuo (6.2 g) and identified as dihydrochloride of 2-hydroxymethy1-2,5diaminopentanoic acid methyl ester by NMR. The ester (6.2 g) is dissolved in 100 ml of absolute methanol and 175 ml of a methanolic solution of sodium methylate (I.I5 g of Na or 5 x 1θ £mole) is added. The reaction mixture is stirred at room temperature under nitrogen for 24 hours. The solvent is evaporated under reduced pressure and the residue is extracted many times with hot chloroform to yield analytically pure 3-hydroxymethyi-3-amino-2piperidone (2.9 g) (yield 81/), m.p. 145'!C.

EXAMPLE 17 2-Ch1oromethy1-2,5-diaminopentanoic acid 3- Hydroxymethy1-3-amino-2-piperidone (2.8 g or 1.94 x 10_a mole) is added to a solution of thionyl chloride (1.4 mi or 1.94 χ 10'2 mole) in anhydrous dimethyl formamide (20 ml). The reaction mixture is stirred at 80°C under nitrogen for 24 hours. Thionyl chloride (1.4 ml or 1.94 x 10‘2 mole) is then added and the stirring continued for 2 hours. The solvent is removed under reduced pressure and the residue is dried under high vacuo overnight, then dissolved in 50 ml of hydrochloric acid (6n) and heated under reflux for 2 hours. The residue obtained after evaporation of the solvent is dissolved in absolute ethanol (100 ml) and is treated with charcoal. After filtration, the final volume of the filtrate is adjusted to 250 ml with absolute ethanol and I9.5 ml of a solution of tri10 ethylamine IN in absolute ethanol is added. The mixture of 2-hydroxymethyl-2,5*diaminopentanoic acid and 2-chloromethyl-2,5-diaminopentanoic acid hydrochloride which precipitates is filtered off and washed with 100 ml of ethanol, 450 ml of chloroform and 250 ml of ether. The filtrate is allowed to stand at 4°C for 24 hours whereupon 2-chloromethy1-2,5-diaminopentanoic acid hydrochioride crystallizes contaminated with a trace of 2-hydroxymethyl2,5-diaminopentanoic acid hydrochloride. The mixture of 2-hydroxymethyl-2,5-diaminopentanoic acid hydrochloride and 2-chloromethyl-2,5-diaminopentanoic acid hydrochloride is redissolved in hydrochloric acid and the same purification procedure is repeated three times. The three batches of crude 2-chloromethyl-2,5-diaminopentanoic acid hydrochloride (1.8 g) are collected and crystallized from water (3.5 ml) and ethanol (17.5 ml) to give 1.3 g of analytically pure 2-chloromethyl-2,5-diaminopentanoic acid hydrochloride (yield 56%), m.p. 14O-142°C. ^7081 EXAMPLE 18 3-Am ino-3-chloromethyi-2-piper idone hydroch lor ide To a solution of 3-hydroxymethyi-3-amino-2-piperidone (7 g or 0.049 mole) in anhydrous dimethylformamide (50 ml) is added one equivalent of thionyl chloride (3-6 ml).

The reaction mixture is stirred at 80°C under nitrogen. After 24 hours, another equivalent of thionyl chloride ,(3.6 ml) is added and stirring is continued for 2 hours. Then the solvent is stripped off under reduced pressure. Trituration of the semi-solid residue with chloroform (2 x 30 ml) leaves 2.1 g of crystalline, analytically pure 3-amino-3-chloromethyl-2-pi per idone hydrochlor i de, m.p. 230°C.

EXAMPLE 19 2-Amino-5-benzy1oxycarbonylaminopentanoic acid A solution of 2,5-diaminopentanoic acid monohydrochloride (16.9 g) and copper carbonate (30 g ) in 750 mi of water is heated at reflux temperature for 4 hours. The insoluble material is filtered off and washed with hot water. The filtrate is allowed to cool to room temperature then sodium bicarbonate (30 g) is added followed by the addition of a solution of benzylchloroformate (25.6 g) in 750 mi of acetone. The reaction mixture is stirred overnight. The precipitate is filtered off, washed with water and then resuspended in hot water (300 ml). Hydrogen sulfide gas is bubbled in the solution for 2 hours.

The solution is then acidified with concentrated hydrochloric acid (7.5 ml) and the black precipitate filtered off. The pH of the filtrate is adjusted to 6 whereupon 2-amino-5-benzyloxycarbonylaminopentanoic acid precipitates 4‘? 0 81 and is collected.

EXAMPLE 20 ct-D i fluoromethyl homocyst i ne Air is bubbled into a solution of α-difluoromethylhomo 5 cysteine (9·25 g or 0.05 M) in 100 mi of water containing a few drops of 1$ aqueous ferric chloride for several hours. The crude α-difluoromethylhomocystine which precipitates is filtered off and then redissolved in water by addition Of concentrated hydrochloric acid. Neutralization of the solution by dropwise addition of concentrated ammonia to a pH of about 4 precipitates a-difiuoromethylbomocystine.

EXAMPLE 21 g-D ifluoromethyl-S-benzyl homocysteine A mixture of α-difluoromethylmethionine (0.2 mole), concentrated hydrochloric acid (0.2 1) and benzylchloride (23 ml) is heated to reflux temperature for 24 hours. The reaction mixture is then concentrated under reduced pressure to a syrup which is dissolved in water. The aqueous solution is extracted with ether, and the aqueous phase is heated with charcoal and filtered. The precipitate is diluted with hot water to a volume of 100 ml and brought to a pH of about 4-5 by addition of aqueous ammonia. On cooling, a-difluoromethyl-S-benzylhomocysteine precipitates and is collected.

EXAMPLE 22 S-Tritylhomocysteine To a solution of anhydrous homocysteine hydrochloride (0.1 mole) in 6o ml of dimethylformamide is added 42 g (0.15 mole) of tritylchloride. The reaction mixture is stirred at room temperature for 2 days. Upon addition of 7081 # sodium acetate solution (500 mlj S-trityl-homocysteine precipitates together with triphenylcarbfnol. Recrystallization in acetone affords pure S-tritylhomocysteine.

EXAMPLE 25 S-Benzyloxycarbonylhomocysteine To an ice cooled solution of L-cysteine hydrochloride (0.1 mole) in 1 molar aqueous sodium carbonate (200 ml) covered with ether (100 ml) is added benzylchloroformate (Ο.Ο95 mole) in one portion with vigorous stirring. After 1 hour at 0°C the temperature is allowed to rise to 10°C and maintained at that temperature for 1 hour. The precipitate is filtered off, washed with water then with acetone and recrystallized from acetic acid to give Sbenzyloxycarbonylhomocysteine.

EXAMPLE 24 -Acetylami no-2-benzyloxycarbonylam ino-2-d i fluoromethyl pentanoic acid To a solution of 1.82 g (0.01 mole) of 5-acetylamino2-amino-2-difluoromethyIpentanoic acid in 22 ml of 1 molar sodium hydroxide is added a solution of 2 g (0.18 mole) of benzylchloroformate in 10 ml of acetone. The reaction mixture is stirred at room temperature for 2 hours and then is carefully neutralized to a pH of 7 using hydrochloric acid whereupon 5-acetylamino-2-benzyloxycarbonylamino-2-difiuoromethylpentanoic acid precipitates and is collected.

EXAMPLE 25 2-Amino-5-acetylamino-2-d ifluoromethy1-1-oxopentaneaminoacetic acid To a solution of 0.25 g (1 mM) of 5-acetylamino-2benzyloxycarbonylami no-2-di fluoromethyl pentano ic ac id 7081 X 50 methyl ester in 4 ml of dioxane is added 0.18 g of benzylglycinate and 0.21 g of dicyclohexylcarbodiimide. The reaction mixture is stirred for 12 hours at room temperature then extracted with ethyl acetate. The aqueous phase is washed with a 10$ solution of bicarbonate then water and dried over magnesium sulfate. Evaporation of the solvent affords an oily residue which is dissolved in glacial acetic acid (5 ml). The solution is then hydrogenated over Pd/C 10$ (30 mg). After stirring the reaction mixture for 12 hours at room temperature the catalyst is filtered off. The filtrate is diluted with toluene and then concentrated under reduced pressure. The residue is purified by ion exchange chromatography on an Amberlite IR 120 active column to give 2-amino-5'acetyiamino-2-di15 fluoromethyl-l-oxopentaneaminoacetic acid.

EXAMPLE 26 2-Acety1amino-5-(2-aminopropionylamino)-2-d i fluoromethy1pentanoic acid ' To a solution of 0.5 g of 2-acetylamino-5-benzy1oxycarbonylamino-2-difJuoromethylpentanoic acid in 4 ml of dimethylformamide is added 0.27 g of benzylbromide and 0.4 ml of dicyclohexylamine. The reaction mixture is stirred for 12 hours and then the precipitate is filtered off. The filtrate is evaporated under reduced pressure, and the residue is partitioned between ethylacetate and water. The organic phase is washed with aqueous hydrochloric acid, water, 5$ sodium bicarbonate, water and brine then dried over magnesium sulfate. The solvent is evaporated and the residue heated in 5 ml of trifluoroacetic acid for 1 hour at room temperature. The excess /170 3 1. trifluoroacetic acid is then stipped off under reduced pressure. The residue is suspended in 5 ml of ether and a solution of N-benzyloxycarbonyl-O-ethoxycarbonylalanine (0.4 g) and triethylamine (0.2 g) in 5 mi of ether is added. Stirring is continued overnight at room temperature. The solvent is evaporated. The resulting syrupy residue is dissolved in glacial acetic acid, and the solution hydrogenated over Pd/C 10$ (20 mg) for 12 hours. The catalyst is then filtered off. The filtrate is concentrated and the residue purified by ion exchange chromatography on an Amberlite IR 120 acidic column to give 2-acetylamino-5-(2-aminoprop ionylami no)-2-d ί fluoromethylpentanoic acid.

EXAMPLE 27 2-Acetylamino-5-am i no-2-d i f1uoromethy1-1-oxopentaneami noacetic acid A solution of 0.25 g of 2-acetylamino-5-benzyloxycarbonylamino-2-difluoromethylpentanoic acid in 4 ml of dioxane is treated at 0°C with 0.11 g of ethylchloroformate and O.lg of triethylamine. The reaction mixture is stirred at 0°C for 1 hour after which 0.18 g of benzylglycinate is added. The reaction mixture is stirred for an additional 12 hours then extracted with ethylacetate.

The organic phase is washed with a solution of bicarbonate, then water and dried over magnesium sulfate. Evaporation of the solvent gives an oily residue which is dissolved in glacial acetic acid (5 ml). The solution is then hydrogenated over Pd/C 10$ (30 mg). After stirring the reaction mixture for 12 hours the catalyst is filtered off. The filtrate is diluted with toluene and concentrated under ,47 08 1 reduced pressure. The residue is purified by ion exchange chromatography on an Amberlite IR 120 acidic column to give 2-acetylamino-5-amino-2-difluoromethyl-1-oxopentaneaminoacetic acid.

EXAMPLE 28 -Acetylami no-2-d? fluoromethyl-2-propionyiaminopentanoic acid methylester To a solution of 0.47 9 5-aC6tyla(!>ino-2-amino-2difluoromethyipentanoic acid methylester in 10 ml of ether Is added simultaneously at 0°C-0.l8 g of propionylchloride and 0.2 g of triethylamine. The reaction mixture is stirred at room temperature for 2 hours then extracted with ethylacetate. The organic phase is washed with a solution of sodium bicarbonate, water, then brine and dried over magnesium sulfate. Evaporation of the solvent gives 515 acetylamino-2-difluoromethyl-2-propionylaminopentanoic acid methylester.

EXAMPLE 29 2-Acetylamino-2-d ifluoromethyl-5-guan id inopentano ic ac id To a solution of 2-acetylamino-5-amino-2-difluoromethylpentanoic acid (0.45 g) in 4 ml of 0.5 M aqueous sodium hydroxide is added 1.8 g of ethylthiouroniumhydrobromide. the pH of the solution is adjusted to 9 with a solution of sodium hydroxide and maintained at that pH for 24 hours. The reaction mixture is then neutralized to a pH of 7 with hydrochloric acid and 2-acetylamino-2-difluoro25 methyl-5-guanidinopentanoic acid is isolated by ion exchange chromatography on an Amberlite IR 120 resin.

EXAMPLE 50 2-Atnino-5-benzyloxycarbonylamino-2-difluoromethylpentanoic acid To a solution of the copper salt of 2-difluoromethyl2,5-diaminopentanoic acid in water, prepared by reacting 2~difluoromethy1-2,5-diaminopentanoic acid monohydrate hydrochloride (2.4 g) with copper carbonate (6 g), is added slowly at 0°C with stirring 1.1 g of benzylchloroformate. The reaction mixture is stirred for an additional 3 hours at room temperature after which hydrogen sulfide is passed through the solution until it becomes colorless. The precipitate is filtered off, and the pH of the aqueous solution is adjusted to 6 by the addition of hydrochloric acid. Upon concentration 2-amino-5-benzyloxycarbonylamino2-difluoromethylpentanoic acid is obtained.

By the above procedure only using tert-butoxycarbony1azide, acetylchloride or benzoylchloride in place of benzylchloroformate gives respectively 2-amino-5-tertbutoxycarbonylamino-2-difluoromethylpentanoic acid, 5acetyiamino-2-amino-2-difluoromethylpentanoic acid and 2-amino-5-benzy1oxycarbony1amίηο-2-d ifluoromethylpentanoic acid.

EXAMPLE 31 2-Acetylamino-5-amino-2-difluoromethylpentanoic acid To a solution of 2.9 9 of 2-amino-5-tert-butoxycarbonylamino-2-difluoromethylpentanoic acid in 10.5 ml of 1 M sodium hydroxide is added at 0°C simultaneously 0.19 g of acetylchloride and 5 ml of 2 M aqueous sodium hydroxide. The reaction mixture is stirred for 3 hours at room temperature. The alkaline aqueous solution is then adjusted 470 81 to a pH of 2 with hydrochloric acid and extracted with ethylacetate. After usual work-up the solvent is evaporated and the residue taken up in trifluoroacetic acid. After concentration and purification by ion exchange chromato5 graphy on a resin 5-amino-2-acetyiamino-2-difluoromethyipentanoic acid is obtained.

EXAMPLE 32 -Amino-2-difl uoromethyl-2-(2-ami nopropionyiamino)pentanoic acid To a solution of 3.2 g of 2-amino-5-benzyloxycarbonyl1θ amino-2-difluoromethyl pentanoic acid in 10 ml of 1 M aqueous sodium hydroxide is added at 0°C simultaneously a solution of tert-butoxycarbonylazide, prepared from 3 9 of tert-butoxycarbonyl hydrazine, and a solution of 5.5 ml of 2 M aqueous sodium hydroxide. The reaction mixture is stirred overnight then extracted twice with 50 ml of ether. The alkaline aqueous solution is then adjusted to a pH of 2 with hydrochloric acid and extracted with ethylacetate. Usual work-up gives a solid residue which is dissolved in 15 mi of dry dimethylformamide and treated at room temperature with 1.7 g of benzylbromide in the presence of 2 ml of dicyclohexylamine. The reaction mixture is stirred for 14 hours and then the precipitate is filtered off. The filtrate is evaporated under reduced pressure. The resulting residue is partitioned between 100 ml of ethylacetate and water. The organic phase is washed successively with 20 ml of 1 normal aqueous hydrochloric acid, 20 ml of water, 20 ml of 5$ aqueous sodium bicarbonate, 20 ml of water and 50 mi of brine then dried over magnesium sulfate. The solvent is evaporated and ^•7081 the residue taken up in 10 ml of trifluoroacetic acid.

After 1 hour at room temperature the excess trifluoroacetic acid is stripped off under reduced pressure and the residue is taken up in a saturated solution of sodium bicarbonate and extracted with 50 mi of ether. The ether phase is dried over magnesium sulfate and then added at 0°C to a solution of N-benzyloxycarbonyl-O-ethoxycarbonylalanine (2 g) in 20 ml of ether. Stirring is continued overnight at room temperature.· The solvent is evaporated and the resulting syrupy residue is taken up in glacial acetic acid (20 ml) and hydrogenated over Pd/C 10% (200 mg). After completion of the hydrogen uptake the catalyst is filtered off. The filtrate is concentrated under reduced pressure with toluene and the residue purified by ion exchange chromatography on an acidic resin to give 5~ amino-2-(2-aminopropionyiamino)-2-d ifluoromethylpentanoic ac id.

EXAMPLE 33 2-[(2,5-Diamino-2-difluoromethyl-l-oxopentane)amino]propionic acid To a solution of 2,5-diamino-2-difluoromethylpentanoic acid monohydrate hydrochloride (2.35 g) in 10 ml of 2 M aqueous sodium hydroxide is added at 0°C simultaneously a solution of 10 ml of 2 molar aqueous sodium hydroxide and a solution of tert-butoxycarbonylazide prepared from 3 9 of tert-butoxycarbonylhydrazine. The reaction mixture is stirred overnight at room temperature and then extracted twice with 250 mi portions of ether. The alkaline aqueous solution is adjusted to a pH of 2 with hydrochloric acid and extracted with ethylacetate. After usual work-up the /470 81 solvent is evaporated and the residue taken up in 40 ml of dry ether. After addition of 1 g of triethylamine an ether solution of 1 g of ethylchloroformate is added slowly at 0°C with stirring. The precipitate is filtered off and 5 the ether solution is added at once to a solution of alanine tert-butylester (1.5 g)· Stirring is continued overnight and the solvent is evaporated. The residue is taken up in trifluoroacetic acid. After concentration and purification by ion exchange chromatography on an Amberlite IR 120 resin 2-[(2,5-diamino-2-difluoromethyl-l-oxopentane)amino]propionic acid is obtained.

Claims (22)

CLAIMS:

1. A compound of the formula Y R HN-(CH_) -C-COR. a 2n | 1 NHRg wherein n is 3 or 4; Y is-CHgF, -CHFg “CFg -CHgCl or CHCI 2 ; R a and R b are independently selected from hydrogen, (C-^alkyl)carbonyl, (Cj_ 4 alkoxy) carbonyl or -CO-CH(NHg)-Rgin which Rgis hydrogen, C-^alkyl, benzyl or p-hydroxybenzyl; and is hydroxy, C|_g alkoxy, -NR^Rg in which R^ and R g are independently selected fran hydrogen and Cj^alkyl, or -NH-CHRg-COOH in which Rg is hydrogen, Cj_ 4 alkyl, benzyl or p-hydroxybenzyl; or a lactam thereof when R & and R^ are each hydrogen and is hydroxy; or a pharmaceutically acceptable salt thereof.

2. A compound as claimed in claim 1 wherein R a and R fa are each hydrogen.

3. A compound as claimed in claim 1 or claim 2 wherein is hydroxy.

4. A canpound as claimed in any preceding claim wherein Y is -CHgF.

5. A canpound of the formula Y, CH 3 -S-(CH 2 ) 2 - C — COR^ wherein Y 2 is -CHgF, -CHFg or -CFg and R^ and R^ are as defined in claim 1, or a pharmaceutically acceptable salt thereof. 4-7081

6. A compound, as claimed in claim 5 wherein is hydroxy.

7. A compound of the formula H 2 N-C(NH)-NH-(CH 2 ) 3 -C-COOH I NHR b wherein Y 2 and R^ are as defined in claim 5, pharmaceutically acceptable salt thereof. or a

8. A compound as claimed in claim 6 or claim 7 wherein R b is hydrogen.

9. A compound of the formula

10. Z. -(CH-)--C-COOH 1 2 2 | nh 2 wherein Z^ is HS-, benzylthio, S-(5*-desoxyadenosin-5-yl) S-methylthio or HOOC-CY 2 (HN 2 )-(CH 2 ) 2 -S-S- and Y 2 Is. as defined in claim 2, or a pharmaceutically acceptable salt thereof. 15 10. A canpound as claimed in claim 9 wherein Z^ is S(5'-desoxyadenosin-5-yl)-S-methylthio.

11. A compound as claimed in any of claims 5 to 10 where in Y 2 is -CHF 2 or - CHjF.

12. A lactam as defined in claim 1.

13. A lactam as claimed in 12 wherein n is 3.

14. 2-Difluorcmethyl-2,5-diaminovaleric acid or a pharmaceutically acceptable salt thereof. 4-7031

15. 2-i'luorcmethyl-2,5-diaminovaleric acid or a pharmaceutically acceptable salt thereof.

16. 2-Chloramethyl-2,5-fiiaminovaleric acid or a pharmaceutically acceptable salt thereof.

17. 2-Difluoromethy1-2,6-diaminocaproic acid or a pharmaceutically acceptable salt thereof.

18. A compound as claimed in claim 1 substantially as described in any of Examples 5 to 33.

19. A pharmaceutical composition comprising a compound as claimed in any preceding claim in association with a physiologically acceptable excipient.

20. A composition according to claim 19 substantially as described in any of Examples 2 to 4.

21. A process for preparing a compound as claimed in claim 6 when R^ is hydrogen, claim 9 when is HS-, or claim 2 when R^ is hydroxy, which comprises treating an amino-protected ester derivative methionine, mercaptoprotected homocysteine, ornithine or lysine with a strong base to form a carbanion intermediate; reacting the intermediate with a halomethyl haloalkylating reagent in an aprotic solvent and, when Y is other than -CHF 2 , in the presence of hexamethylphosphortriamide, at a temperature of -120° to 120°C for from 30 minutes to 48 hours; and subjecting the product to acid or base hydrolysis.

22. A process for preparing a ccmpound as claimed in claim 10, which comprises reacting 5'-p-toluenesulfonyladenosine, 5'-chloroadenosine or 5'-bromoadenosine, each optionally protected as the 2',3'-isopropylidene, wit-h a sodium or lithium salt of an a-halamethylhamocysteine to form the S-adenosyl-a-halcmethylhanocysteine derivative; hydrolysing the derivative with acid to remove any 2',3'-isopropylidene protecting groups; and treating the product with methyliodide in an acidic solvent.