NZ187538A - 2-amino-2-halomethyl-3-phenylpropionic acid derivatives - Google Patents

Description

New Zealand Paient Spedficaiion for Paient Number 1 87538 f 875 "i * S> AMENDED under Section of the Patents Act 1953 frora/.^ ASSISTANTCOMMISSIONER OF PATENTS Priority Dat#(s): 1^.1.11 Complete Specification Filed; cisst! ■ iQ^i v •«• < tit IJ i«< i P.O. Journal, No: .

No.: Date: <^1 C-2 ^-7^Z.o C^1C5b^ M C=0"D^<^\t£l= NEW ZEALAND PATENTS ACT, 1953 complete specifications amended 187538. Merrell Toraude Et Compagnie. x-Halomethyl Amino acids. Complete specificaton amended under section 14 and section 16.

COMPLETE SPECIFICATION " a-HRICVEEiXL AMKD ACIDS" Ic/We, MERRELL TORAUDE ET COMPAGNIE a corporation organized under the laws of France, having an office at 16 rue d'Ankara, 67000 Strasbourg, France hereby declare the invention for which £ / we pray that a patent may be granted to naft/us, and the method by which it is to be performed, to be particularly described in and by the following statement: - 1 - (' followed by page \a) f S - - .3 s -Mt-685- . FIELD OF INVENTION This invention relates to novel pharmaceutica11 y useful a-halomethyl amino acid derivatives which are inhibitors of aromatic amino acid decarboxylase.

BACKGROUND OF INVENTION The amino acids tryptophan, 5-hydroxytryptophan, 3,4-dihydroxypheny1alanine (DOPA), tyrosine and phenylalanine are metabolica11y converted to tryptamine, 5-hydroxy-tryptamine, 3,4-dihydroxyphenethylamine or dopamine, tyra-10 mine and phenethylamine respectively by an aromatic amino acid decarboxylase. It is believed that the aromatic amino acid decarboxylase enzyme is non-specific, particularly, insofar as peripheral catalysis is concerned. Evidence does exist, however, to indicate that in the 15 brain specific decarboxylation enzymes exist for each of DOPA and 5-hydroxytryptophan.

The above-enumerated aromatic amines are known to be involved in various pathophysiological processes. For example, it has been found that tryptamine, the decarboxy-20 lation product of tryptophan is enzymatica11y methylated to monomethyltryptamine which in turn is methylated enzymati-cally to dimethyltryptamine (DMT) in human red blood cells, plasma and platelets. The methylating enzyme is present in many mammalian species and has been shown to be produced 25 in brain tissues of several species including man. DMT which has strong hallucinogenic or psychomimetic properties may play a role in the etiology of schizophrenia and other psychotic disorders. Hence any agent which would block formation of DMT may be useful as an antipsychotic agent. -lo- rspyj , r- -> Blocking the decarboxylation of tryptophan results in decreased levels of tryptamine, removing the substrate for DMT formation. Hence an inhibitor of aromatic amino acid decarboxylase which would block conversion of tryptophan 5 to tryptamine may be useful as an antipsychotic agent.

Both 5-hydroxytryptamine (5-HT), the decarboxylation ^ product of 5-hydroxytryptophane, and 3,k-dihydroxyphenethy1 - amine (dopamine) the decarboxylation product of DOPA are involved in peripheral and central physiological processes, 110 and agents which are effective in the control of levels of these amines have resulted in useful pharmacological agents. It has been shown that central or brain levels of 5-HT and norepinephrine, which is formed metabolica11y by hydroxyla-tlon of dopamine, are higher in patients with manic dis-15 orders than in individuals without such disorders. It has also been shown that agents which decrease central levels of monoamines, for example, 5-HT and particularly norepinephrine have antimanic properties when given to human subjects, whereas drugs that increase monoamine levels could ^20 precipitate mania in susceptible individuals. Hence, agents which block formation of 5-HT and dopamine, such as, for example, by inhibiting the aromatic amino acid decarboxylase enzyme which converts 5-hydroxytryptophan and DOPA to 5-HT | and dopamine respectively may be useful as antipsychotic agents or major tranquilizers in treating manic disorders.

It has also been shown that agents useful in inhibiting the decarboxylation of DOPA to dopamine are useful in the treatment of Parkinsonism when administered concurrently with exogenous DOPA or L-DOPA. It is believed that 30 Parkinsonism is due, at least in part, to decreased central levels of dopamine since exogenous administration of DOPA or L-DOPA is known to be an effective means of treating Parkinsonism. However, since exogenously administered DOPA is readily converted enzymatical1y to dopamine peripherally it is necessary to administer large amounts in order to have .increased absorption centrally. DOPA readily penetrates the blood-brain barrier whereas dopamine does not. Administration of DOPA or L-DOPA in conjunction with a peripherally active inhibitor of the enzyme which converts DOPA to dopamine reduces the amount of L-DOPA that must be administered in order to have adequate circulating levels for central absorption. Other advantages are also realized by administration of an aromatic amino acid decarboxylase inhibitor along with L-DOPA. By preventing formation of dopamine peripherally, side effects attributed to dopamine such as, cardiac arrhythmia, nausea and vomiting may be avoided.

Studies indicate that levels of 5-hydroxy tryptarni ne (5-HT) are lower in patients with depressive syndromes than in individuals without such syndromes. Also, administration of exogenous L-5-hydroxytryptophan (L-5-HTP) is effective in treating certain depressed patients. However, as with DOPA, since L-5-HTP is readily metabolized peripherally to 5-HT it is necessary to administer large amounts of L-5-HTP in order to achieve increased central levels of the amino acid. It has been shown that by administering an inhibitor of the aromatic amino acid decarboxylase enzyme that catalyzes the formation of 5-HT from 5-HTP peripherally the amount of exogenous 5-HTP required to give increased central levels is markedly ^ <=7 - • /?, V.vy /' ' .y -m-=-885"~~ reduced. In other words inhibitors of aromatic amino acid decarboxylase when used in conjunction with exogenous 5-HTP have been shown to be useful in treating depression.

Agents which block peripheral conversion of 5-HTP to 5-HT may be useful in treating other conditions which respond to increased central levels of 5-HTP as a result of exogenous administration of 5-HTP. It has been shown that exogenous L-5-HTP is useful in treating action myoclonus. Also, studies reveal that administration of exogenous 5-HTP is useful in treating insomnia. Hence concurrent administration of 5-HTP and an aromatic amino acid decarboxylase inhibitor may be beneficial in treating these conditions.

Blocking peripheral formation of 5-hydroxytryptamine may result in other beneficial effects since it is known that 5-HT is involved, for example, in the etiology of rheumatoid arthritis arid the carcinoid syndrome by increasing collagen levels. Also, it is reported that 5-HT Is the primary autocoid responsible for anaphylactoid reactions in human subjects as well as bronchoconstriction In asthmatic human subjects, and agents which antagonize or inhibit formation of 5-HT are useful in treating these conditions. 5-HT is known to cause platelet aggregation and has been implicated as a causal factor in the postgastrectomy dumping syndrome and migraine headache. Methyl-sergide, a 5-hydroxytryptamine antagonist, has proven effective in treating post-gastrectomy dumping syndrome.

It has been suggested that phenethylamine, the decarboxylation product of phenylalanine, as an endogenous compound contributes to schizophrenic symptoms and triggers 1875 migraine headaches. Also, it has been suggested that endogenous tyramine, the decarboxylation product of tyrosine, contributes to seizure disorders.

Hence, it is readily evident that agents which are useful in regulating the levels of aromatic amino acids and amines find use in many pharmacological situations. The compounds of the present invention are inhibitors of the aromatic amino acid decarboxylase which converts tryptophan, 5-hydroxytryptophan, 4~dihydroxyphenyla la-nine, tyrosine and phenylalanine to the respective amines and hence provide useful pharmacologic agents.

The compounds of the present invention are represented by the following general Formula: ^' 4 t%6 NHK i In the above general Formula 1 Y is FCH2~ or F2CH-, R-^ is hydrogen, alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and is straight or branched, or wherein R27 is hydrogen, a straight or branched lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenz-yl; R2 is hydroxy, a straight or branched a 1koxy group and Re is hydrogen or a straight or branched alkyl group SUMMARY OF INVENTION Formula I NH2 of from 1 to 8 carbon atoms, -NR7Re wherein each of R7 aeT7 -m-h-805— of from 1 to 4 carbon atoms, or -NHCH-COOH wherein Rg is Rs hydrogen, a straight or branched lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzyl; |^ach of R3, R4, Rs, R'4 and Rs has the meaning defined in the fol-5h.^* lowing Table I wherein Rio is hydrogen, -a straight or— ^..-branched alkyl group of from 1 to 8 Crirhnn atoms,, alkyl-carbonyl wherein the alkyl moiety is straight or branched and has from 1 to 6 carbon atoms, benzoyl or phenylalkyl-enecarbonyl wherein the alkylerie moiety is straight or 10 branched and has from 1 to 6 carbon atoms-.

TABLE I R3 R4 Rs R1 4 Re H -0-CH2-0~" H H —h h h h h . H H OR 10 H H %y°°° ^ nlvV-' H OR 10 H H H svp'.y H OR 10 OR 10 H H OR10 H CI H H H OR10 CI H H CI OR10 H H H CI OR10 Cl H H -c 1, f h oftto h h —eh :—h H h GHs —04 H QJ H -C44© —H N C-J-7-f ti CHe OR la H CH3 H CH3 C1 H CH3 H CH3 c,, -+1—1 H OR-ro H &H3 £V- ~H H OR-1-0 H CzHs f-- Xv-V' m NOW AMENDED M+-685" < O ?• %> %• >?«*■ 3-V" -H- -H- -tert-C+Hn. -t-er-t-C4 H-s Pharmaceut ica 11 y acceptable salyts and individual optical isomers of the compounds of general Formufa I are also included within the scope of/this invent/on.

The compounds of general Formula \ are useful pharmacological agents in that s4id compounds are inhibitors of aromatic amino acid deca/boxylase ana useful as intermediates In the preparation of useful pharmacological agents.

DETAILED DESCRIPTION OF INVENTION In the above general Formula I the term a 1kylcarbony1 is taken to mean the group alk/l-C- wherein the alkyl moiety ) has from 1 to 6 cj&rbon atoms pnd is a straight chain or branched chain.

The term^benzoyl as u^ed in general Formula I means 0 the group —C-.

The tfsrm phenylalkyylenecarbonyl as used in general 0 Formula J is taken tcymean the group (s~/)—alkylene-C-wherein/the alkylene/moiety has from 1 to 6 carbon atoms and i^/a straight rniain or a branched chain, illustra- OR i o H C2H5 H c2h h OR i o h ORio h h ORio ORi 0 ORi 0 h j h h OCH3 OH h ORio ORio h h h ORio h h h H, t875"<0 JO but excluding compounds wherein (a) Y=CH2F; R2, R3, R'4, and Rg=H; R2=-OH or alkoxy; and r4 and R5 both are -0R1Q, wherein R1Q is H or alkylcarbonyl, (b) Y=CH2F; Rx, R3, R4, R'4, and Rg=H; R2 is -OH or alkoxy; R =-0H, or (c) Y=CH2F; Rx, R3, R'4, R5, and Rg=H; R2 is -OH or alkoxy; and R4=-OH, Pharmaceutica 11 y acceptable salts and individual optical isomers of the compounds of general Formula 1 are also included within the scope of this invention.

The compounds of general Formula 1 are useful pharmacological agents in that said compounds are inhibitors of aromatic amino acid decarboxylase and useful as intermediates in the preparation of useful pharmacological agents.

DETAILED DESCRIPTION OF INVENTION In the above general Formula I the term alkylcarbonyl 0 is taken to mean the group alkyl-C- wherein the alkyl moiety has from 1 to 6 carbon atoms and is a straight chain or branched chain.

The term benzoyl as used in general Formula 1 means 0 the group \^/)— The term phenyla 1 kylen-ecarbony 1 as used in general 0 /—A " Formula I is taken to mean the group (\//—alkylene-C- wherein the alkylene moiety has from 1 to 6 carbon atoms and is a straight chain or a branched chain, illustra- patent of^c? 2 3 NOV 1982 received -7- 1 87 ~ 7 -M~68*r tively, methylene, ethylene, isopropylene and butylene.

Illustrative examples of straight or branched alkoxy groups having from 1 to 8 carbon atoms as used herein are methoxy, ethoxy, isopropoxy, n^-butoxy, tert-butoxy. ji-penty 1 oxy, tert-pentoxy, ri-hexyloxy and ri-octyloxy.

Illustrative examples of straight or branched alkyl groups having from 1 to 6 carbon atoms are methyl, ethyl, r^-propyl, isopropyl, jn-butyl, tert-buty 1 and jn-pentyl.

Illustrative examples of pharmaceutica11y acceptable salts of the compounds of this invention include nontoxic 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 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 I I I A, for example, aluminum, organic amines, such as, primary, secondary or tertiary amines, for example, cyclohexylamine, ethylamine, pyridine,^methylamino-ethanol^ ■'£thanolamine and piperazine. The salts are prepared by conventional means.

Preferred compounds of this invention are those of general Formula I wherein F>! is hydrogen or alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and Is straight or branched^with compounds wherein Ri is jjydrogen being more preferred. Another preferred embodiment of this invention is the compounds of general Formula wherein R2 is hydroxy or a straight or branched alkoxy - ^H-=8S5" group of from 1 to 8 carbon atoms. Compounds wherein R2 is hydroxy are more preferred. Compounds of general Formula I wherein each of R3, R4, Rs> R'4 and Ra is hydrogen or ORio wherein Ri0 is hydrogen represent another preferred embodiment of this invention. Compounds of general Formula I wherein Y is FCH2- or F2CH- are also preferred.

Illustrative examples of compounds of general Formula I are the following-. 2-difluoromethy1-2-amino-J-phenylpropionic acid, 2-d i f 1 uoromethyl -2-ami no-3- (3-h'ydroxypheny 1) propion ic ac id, 2-d ifluoromethy1-2-amino-3-(3,4-dihydroxyphenyl)propion ic acid, 2-d i fluoromethy1-2-ami no-J-(4-hydroxyphenyl)prop ion ic ac id, 2-fluoromethy1-2-am i no-J-(4-ch1oro-2-hydroxyphenyl)pro-pionic acid, V ?■ I."'-' pionic acid.

. • Jj-d ichlorometh \V"" prtrpTonic acid, ino-3-(2-chloro-3-benzoyloxyphenyl)- < 2-fluoromethy1-2-amino-3-(2,4-d ichloro-3-hydroxypheny1)-propionic acid, 2-d i fluoromethyl-2-amino-3-(2-chloro-4-hydroxyphenyl)pro-pionic acid, 'g-chloromethy 1 -2-amino-3- (2-ch4oro-6-methyTpheny-l ) propiorH^-25 ao id 2-fluoromethy1-2-amino-3-(2,4-d ichloro-6-methy1pheny1)-propionic acid, ■fi-diehloroinetliyl 2 omino 3 ('I chloro6 methyl phcnyl)prg pionic acid, 30 2-d ich loromethy I-^am-i-preprOnTc ac idj ^3-T2-hydroxy-4,6-d imethylphenyl)- 85~ ■a-chloromathyl -2»am i no-3- (2-^Mo-flQ--4-j-6---d-i-methy 1 pheny 1-)-propionic acidj--~ 2-fluoromethy1-2-am ino-3-(4-hydroxy-6-methy1 phenyl)propion ic ac i d, 2-d i f1uoromethy 1-2-amino-3-(4,6-d iethy1-2-hydroxypheny1) ■ propionic acid, ac id. 2-fluoromethy1-2-amino-3-(4-chloro-6-tert-butylphenyl)-propionic acid, E-d i f 1 ueromethyl-2-ami-n&-3~(6-tert-but y 1 -4-hydroxy phen y-1--)-,-pro picnic acid,— 2-d ich loromethy 1-2- (N-ethoxycarbony lam ino)-3- (4-ri-butoxy-phenyl)propionic acid, N,N-di-j2~ propyl 2-d if luoromethy 1 - 2-ami no-3- (4-acetyloxy-phenyl)propionamide, 2-fluoromethy1-2-[N-(2-amino-1-oxoethy1)amino]-3-(3-hydroxyphenyl)prop ionic acid, 2-f1uoromethy1-2-amino-3-(3j 4-d i hydroxy)phenyl-1-oxo-propylaminoacetic acid, 2-[(2-difluoromethy1-2-amino-l-oxo-3-phenyl)propyl amino)-dihydrocinnamic acid, 2-d i f1uoromethy1-2-(1-oxoethy1 am i no)-3-(4-hydroxy)pheny1-1-oxopropyl amino-2-prop ionic ac id, methyl 2-fluoromethyI-2-(1-oxoethylamino)-3-(4-hydroxy)-pheny1-1-oxopropyl am i noacetate, ■0 chloromcfrhyl ■ 0 amino 3 phenylpropionomidcj— N,N-d imethyl 2- d i f1uoromethy1-2-amino-3-(3-hydroxypheny1) J44~885~ propionamide, NjN»diethyl 2-dichloromethy1-£•amino-3*(3114'-d imothoxy •phenyl)propionamidc> ■ N-ji-butyl 2-d i f 1 uoromethy 1 - 2-am i no-3 - (4-hydroxy pheny 1) -propionamide, -t-sopropy 1—2^-i-ch-] rox^yphony 1) " •prop i onager-. tert-buty1 2-fluoromethy1-2-ami no-3-(4-hydroxypheny1)-propionate, ethyl-2-d i fluoromethy1-2-amino-3-(4-chloro-3-methoxyphenyl)-propionate, and 2- fl uorome thy 1 -2-ami no-3- (4-'hydroxy phenyl) propionamide.

The compounds of general Formula I are irreversible inhibitors of the enzyme which metabolica11y catalyzes the conversion of tryptophan, 5-hydroxytryptophan, 3.>4-di-hydroxyphenylalanine, tyrosine and phenylalanine to tryp- P~C \ tamine, 5-hydroxytryptamine,^ 3* 4-d ihydroxyphenyl/ethylamine, tyramine and phenethylamine respectively. As indicated hereinabove results of studies indicate that the enzyme responsible for the conversion of the above-enumerated amino acids to the respective amines peripherally is a non-specific aromatic amino acid decarboxylase. For central conversion studies indicate that specific decarboxylases are responsible for the conversion of each of 5-hydroxytryptophan and 3,4-d ihydroxypheny1 a 1 an ine whereas the remaining above-enumerated amino acids are enzymatica11y transformed to the respective amines by a non-specific aromatic amino acid decarboxylase. The compounds of the present invention are effective in irreversibly inhibiting both centrally and peripherally the activity of non-specific ; ^ !^7r'^ • / >, , ;/ aromatic amino acid decarboxylase as well as the activity of 3,4-dihydroxyphenylalanine (DOPA) decarboxylase. As used herein with regard to the utility of the compounds of the present invention the term central refers to the central 5 nervous system, primarily the brain, whereas peripheral refers to other body tissues wherein the decarboxylase ^ enzyme is present. Selectivity of inhibition of the amino acid decarboxylases centrally or peripherally by administering compounds of general Formula I is dose dependent. 10 As irreversible inhibitors of aromatic amino acid ^ decarboxylase, and DOPA decarboxylase the compounds of the present invention possess many pharmacological utilities. As peripheral irreversible inhibitors of aromatic amino acid decarboxylase the compounds of general Formula I are 15 useful in the treatment of Parkinsonism when given in conjunction with 3,4-dihydroxypheny1 a 1 anine (DOPA) or L-3,4-dihydroxyphenylalanine (L-DOPA). DOPA and more particularly the active isomer L-DOPA are known to be effective in treating Parkinsonism when administered 20 systemically, usually in an amount from 0.5 to 1 gram daily initially after which the amount administered is gradually increased over a 3 to 7 clay period to a maximally tolerated daily dose of about 8 grams. Concurrent administration of a compound of general Formula I and L-DOPA provides 25 an improved method of treating Parkinsonism in that the compounds of Formula I will block the decarboxylation of L-DOPA to L-3,4-dihydroxyphenethylamine (L-dopamine) peripherally by inhibiting the activity of aromatic amino acid decarboxylase enzyme, thus retaining high circulating 50 levels of L-DOPA for central absorption and also preventing ^ <7/ r 1 '7! (Sfr v." '■ -j peripheral formation of increased levels of dopamine which Is known to result in certain undesirable side effects such as cardiac arrhythmia. By concurrently administering a compound of general Formula I and L-DOPA the amount of L-DOPA administered may be reduced 2 to 10-fold as compared to amounts required for utility when L-DOPA is administered alone. It is preferred that the compounds of this invention be administered prior to administration of L-DOPA. For example, a compound of Formula I may be administered from 30 minutes to 4 hours prior to administration of L-DOPA depending on the route of administration and condition of the patient to be treated.

The compounds of general Formula I are also useful In treating depressive syndromes in individuals when given in conjunction with 5-hydroxytryptophan (5-HTP) or more particularly the active levo isomer which is known to be useful in the treatment of depression when administered systemica11y. The compounds of general Formula I, by inhib iting peripherally the activity of aromatic amino acid decarboxylase will block the conversion of 5-hydroxytryptophan to 5-hydroxytryptamine thus retaining higher circulating levels of 5-HTP for central absorption. The compounds of general Formula I when administered concurrently with exogenous 5-HTP are also useful in treating action myoclonus which is known to be effectively treated by increasing central levels of 5-HTP.

The compounds of general Formula I, by virtue of their Inhibitory action on aromatic amino acid decarboxy 1ase peripherally are also useful in the treatment of rheumatoid arthritis, carcinoid syndrome, anaphylactoid reactions in humans, bronchoconstriction in asthamatic humans as well as other conditions known to be caused by high peripheral levels of 5-hydroxytryptamine.

As indicated hereinabove it has been shown that agents which decrease the elevated levels of 5-HT and norepinephrine, the hydroxylation product of dopamine, are useful in treating patients with manic disorders. Hence, as central irreversible inhibitors of aromatic amino acid decarboxylase, and DOPA decarboxylase the compounds of general Formula I are useful in treating manic disorders. Add it ionally, by virtue of the central inhibitory action of the compounds of general Formula I on aromatic amino acid decarboxylase said compounds may also be useful as antipsychotic agents, since central levels of tryptamine are decreased, and useful in the treatment of schizophrenia and seizure disorders since central levels of phenethylamine and tyramine are decreased by administration of a compound of general Formula I.

The utility of the compounds of general Formula I as irreversible inhibitors of aromatic amino acid decarboxylase may be demonstrated as follows. A compound of general Formula 1 is administered as an aqueous solution or suspension to rats or mice. At different time intervals after administration of the compound from 1 to 48 hours the animals are sacrificed by decapitation and aromatic amino acid decarboxylase activity is measured by a radiometric assay as described by Christenson et al.> Arch. Biochem. Biophys. l4l, 356 (1970) in homogenates of kidney, heart and brain prepared according to Burkard et al., Arch.

Biochem. Biophys. 10J, I87 (1964). t 87r The compounds of this invention can be administered in various manners to achieve the desired effect. The compounds can be administered alone or in the form of pharmaceutical preparations to the patient being treated either orally or parenterally, for example,- subcutaneous 1y, intravenously or intraperitonea11y. The compounds can be administered by intranasal instillation or by application to mucous membranes such as that of the nose, throat and bronchial tubes, for example, in an aerosol spray containing small particles of a novel compound of this invention in a spray solution or dry powder form.

The amount of novel compound administered will vary and can be any effective amount. Depending on the patient, the condition being treated and the mode of administration, the quantity of novel compound administered may vary over a wide range to provide an effective amount in a unit dosage,form. When the compounds of general Formula I are administered to affect a peripheral ^ ^ ,y irreversible inhibition of aromatic decarboxylase the effective amount of compound administered will vary from about 0.1 mg/kg (milligrams per kilogram) to 100 mg/kg of body weight of the patient per dose and preferably from 5 about 5 mg/kg to 25 mg/kg. For example, the desired peripheral effect can be obtained by consumption of a unit dosage form, such as, for example, a tablet containing from 10 to 250 mg of a novel compound of this invention taken 1 to 4 times daily. When the compounds of general 10 Formula I are administered to achieve a central irreversible inhibition of aromatic decarboxylase or 3,4-dihydroxy-phenylalanine decarboxylase the effective amount of compound administered will vary from about 100 mg/kg to 500 mg/kg of body weight of the patient per day and pre-15 ferabiy from about 150 mg/kg to 300 mg/kg. For example, the desired central effect can be achieved by consumption of a unit dosage form, such as, for example, a tablet containing from about 350 mg to 500 mg of a novel compound of this invention taken from 1 to 4 times daily. 20 As used herein the term patient is taken to mean warm blooded animals such as mammals, for example, cats, dogs, rats, mice, guinea pigs, sheep, horses, bovine cows, and humans.

The solid unit dosage forms can be of the conven-25 tional 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 and inert fillers such as lactose, sucrose and corn starch. In another embodiment, the novel compounds <Mb*885 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 add i t ion 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 general 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 subcutaneous 1y or intramuscularly as depot injections or implants. Implants may employ inert materials such as biodegradable polymers or synthetic silicones, for exampl Silastic, silicone rubber manufactured by the Dow-Corning Corporation. 1 H-T V. J For use as aerosols the novel compounds in solution or suspension may be packaged in a pressurized aerosol container together with a gaseous or liquified propellant, for example, dichlorodif1uoromethane, dichlorodif1uoro- '1,1- methane wi th jtd i chl oro^i i f 1 uoroethane, carbon dioxide, &S- nitrogen or propane, with the usual adjuvants such as eosolvents, and wetting agents, as may be necessary or desirable. The compounds may also be administered in a non-pressurized form such as in a nebulizer or atomizer. 10 As indicated hereinabove the compounds of general Formula I find particular utility when administered together with exogenous L-DOPA in which case individual formulations of a compound of general Formula I and L-DOPA may be administered, or both active ingredients may be formulated 15 into a single combination pharmaceutical formulation. In either mode of administration the amount of compound of general Formula I as compared to the amount of L-DOPA administered will vary from about 1:1 to 1:10. A combination formulation may contain an internal portion con-20 taining L-DOPA and an outer portion containing a compound of general Formula I, each active ingredient being suitably formulated. A particularly suitable combination formulation may be prepared by compressing L-DOPA, optionally with suitable carriers, to a core, providing 25 said core with a laminated coating that is resistant to gastric juice, and applying over the coated core an external layer that contains a compound of general Formula I suitably formulated. Using such a combination formulation the decarboxylase inhibitor, that is, a 30 compound of General Formula I is released, preferably t 8753 -m-885* to 60 minutes prior to the L-DOPA. The laminated coating may be formed by use of a nonaqueous solution of glycerides or a water-insoluble polymer such as ethyl wherein the L-DOPA is enteric coated by use of mixtures of shellacs and shellac derivatives and cellulose acetate phthalates may also be enployed.

In the specific examples included hereinbelow illustrative examples of suitable pharmaceutical formulations are described.

In addition to being useful pharmacological agents compounds of general Formula I are also useful as intermediates for the preparation of useful cephalosporin anti biotics. Compounds of general Formula I wherein R2 is hydroxy are useful in the preparation of cephalosporin derivatives of the following general Formula II: In the above general Formula II, y, Ri, R3j R4, R5j R>4 and Rs have the meanings defined in general Formula I; M is hydrogen or a negative charge; and X is hydrogen or acetoxy.

The compounds of general Formula II and the pharma-ceutically 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, cellulose or cellulose acetate phthalate. Formulation C00M Formula I I 1 JO~88T FT ■tj/ cephalexin, cephalothin, or cephalog1ycine. The compounds of general Formula II and pharmaceutically acceptable salts and isomers thereof can be administered alone or In the form of pharmaceutical preparations either 5 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 pills or in the form of 10 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 15 general Formula II, 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 pharmaceut!ca11y acceptable salts and individual optical isomers thereof SO are active are Staphylococcus aureus, Salmone1 la schot-) muehleri, Klebsiella pneumoniae, Diplococcus pneumoniae and Streptococcus pyogenes.

Illustrative pharmaceutical1y acceptable non-toxic . ^ inorganic acid additions salts of the compounds of general Formula II 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 ascorbata. The 30 salts can be formed by conventional means.

K. 'to ?• & S. <vz~ ,/ • VLU3&T~ | now AMENDED | Illustrative examples of cephalosporin derivatives /fcKfLuorofnel as represented by general Formula II are 7-[[2/^cotyluiit^ ' 2-ami no-3-phenyIpropionyl]ami no]-3-acetyloxymethy1-8-oxo- ,%\.. .5-thia-l-azabicyclo[ 4.2.0]oct-2-ene-2-carbo>/yl ic acid. di'-f Uujrofn-ei^i I / 7-[ [2-^accty 1 ona-2-ami no-3-(3-hydroxypheny 1Yprop iony 1 ] -ami no]-3-acetyloxymethy1-8-oxo-5-th ia-l-aiab ieyelo[4.2/0] - ell -pLtxoro(r\c"rt\y I 3-)0C0t ',/l ono-2-am i n< IP' oct-2-ene-2-carboxyl ic acid, 7-[ [g^acotv/l ono-2-amTno-(3,4-di hydroxypheny1)prop iony1]ami no]-3-acetyloxyme/hy1 -8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2/ene-2-carbox/lic r-fluorometk^ /, / . acid, and 7-[ [2-jacoty 1 orre-2-ami no-3y( 4-hydroxyphe/ny 1) -propionyl]ami no]-3-acetyloxymethyl /8-oxo-5-thiayl-aza-b icyc1o[ 4.2 .0] oct-2-ene-2-carboxy<l ic ac i d The compounds of general Formula II wherein is hydrogen are prepared by coupljmg 7-ami nocep/ia 1 osporan ic 15 acid or a derivative thereof jnaving the formula H2N </H2X / Formula I I I COO} wherein X and M have the meanings defined in general Formula II with an ac i/d of the formula R4 R3 R5~0/H2"f-cooH !». (/- NHS Formula IV of a functiona1 /derivative thereof such as the acid chloride or an/acid anhydrid^ and in the presence of a dehydrating agent/such as dicyc/ohexylcarbodiimide, when the ! / free acid is used, wherein r3, r4, r5, r>4 and Re have the meanings defined in general Formula II and the amino group .1875 ' .' ASAMENDED,- Illustrative examp1es of cepha1osporin derivatives as represented by general Formula II are 7-[[2-difluoromethyl-2-ami no-3- pheny 1 pr op i ony 1 ] ami no] -3 -acetyl oxymethy 1 -8-oxo-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxy1ic acid/ 7-[[2-difluorcnet±iyl-2-amino-3- (3-hydroxypheny 1) prop iony 1 ] - ami no]-3-acetyloxyme thy 1-8-oxo-5~ thia-1-azab icyclo[4.2.0]- . oct-2-ene-2-carboxy 1 ic acid, and 7-[[2-difluoramethyl-2-aniino-3-(3j4-dihydroxypheny1)propiony1]ami no]-3-acetyloxymethy1 -- 8-OXO-5-thia-l-azabicyclo[4.2.0]oct-2-ene-2-carboxy1ic acid.

The compounds of general Formula II wherein Rt is hydrogen are prepared by coupling 7-aminocepha1osporanic acid or a derivative thereof having the formula CH2X Formula II COOM wherein X and M have the meanings defined in general Formula II with an acid of the formula Rs //—CH2-C-C00H Formula IV m 2 or a functional derivative thereof such as the acid chloride or an acid anhydride and in the presence of a dehydrating agent such as dicyc1ohexy1carbodiimide, when the free acid is used, wherein R3, R4, R5, R'4 and Rs have the meanings defined in general Formula II and the amino group 1 87-7 3 Is protected with a suitable blocking group such as tert-butoxycarbony 1 followed by acid hydrolysis to remove the amino protecting groups.

The coupling reaction is genera 11y 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°C 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 general Formula IV are prepared by procedures er described hero-inabove and the compounds of Formula II) are commercially available or are made by procedures well known in the art.

The compounds of general Formula II wherein Ri is other than hydrogen are prepared from the corresponding derivatives wherein Ri is hydrogen by the general procedures set forth hereinbelow for compounds of general Formula I wherein Ri is other than hydrogen.

The compounds of general Formula I wherein Ri is hydrogen, R2 is hydroxy, both R3 and R4 are 0Rio wherein Rio is hydrogen or both R4 and Rs are ORio wherein Ri0 is hydrogen or both R4 and Rs together are -0-CH£-0- or wherein each of R3, R4, Rs, R'4 and Rs has the meaning otherwise defined in Table I except Ri0 is methyljare prepared by treating a suitably protected phenyl propionate of the formula CXfv R12 R11 Formula V 187533 - 23 - . _ , v ^ ^ -j with a strong base to generate a carbanion which is treated with a suitable halomethyl alkylating reagent followed by (I) acid hyrolysis and, if necessary to cleave the aromatic methyl ether group, treatment with hydrogen bromide in water or acetic acid for 4 to 24 hours at 25°C to 125°C or (II) treatment with hydrogen bromide in water or acetic acid for 4 to 24 hours at 25°C to 125 C. in the above general Formula V is a straight or branched alkoxy group having from 1 to 8 carbon atoms, R^ is hydrogen, phenyl, a straight or branched alkyl group of from 1 to 8 carbon atoms, methoxy or ethoxy; Rc is phenyl or a straight or branched alkyl group of from 1 to 8 carbon atoms; or R^ and Rc taken together form an alkyl-ene group of from 5 to 7 carbon atoms, that is, -CH2-(CH2)m-CH2- wherein m is an integer of from 3 to 5» I 1lustrative examples of straight or branched alkyl groups of from 1 to 8 carbon atoms which R^ and Rc may represent are methyl, ethyl, ri-propyl, isopropyl, n-butyl, tert-butyl, n-hexyl, ri-octyl and neopentyl. Each of Rn, Ri2, R13, R'12 and Ri4 has the meaning defined in the following Table II: table i i ri i r12 r i; i p Km -opc-o- h h h h3c nch3 h -0-c-0- h h h3c ch3 h ch2-o- h h h h och3 h h h och3 h h h h och3 och3 h h och3 h ci h h ^ y PATENT OFPCa ^^ecavEQ^ 137533 h OCH3 ci h h ci och3 h H h ci och3 cl . H h 0ch3 H ch3 h ch3 OCH3 h C2H5 • H C2hs H OCH3 H OCH3 H H och3 OCH3 OCH3 H H h och3 OCH2Ph h OCH3 OCH3 H h H OCH3 H H H H h H cl H c2h5 H H cl H tert-C 4H9 H h och3 H tert-C4Hs Suitable strong bases which may be employed in the 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, an alkyl lithium, for example, butyl lithium or phenyl lithium, a lithium di-alkylamide, for example, lithium diisopropylamide, or lithium amide, tertiary potassium butylate, sodium amide, metal hydrides, for example, sodium hydride or potassium hydride, tertiary amines, such as, triethylamine, lithium acetylide or dilithium acetylide. Lithium acetylide, di-lithium acetylide, sodium hydride and lithium diisopropyl- * t 87''' -=-Mt^685" amide are particularly preferred bases.

Suitable halomethyl alkylating reagents which may be employed in the above reaction are illustratively chloro-fluoromethane, bromof1uoromethane, fluoroiodomethane, 5 chlorodifluoromethane, bromodif luoromethane} difluoroiodomethane, bromochloromethane, dichloromethane, chloroiodo-methane, bromod ichloromethane and d ichloro iodomethane. The halomethyl alkylating reagents are known in the art.

The a 1kylating reaction may be carried out in an 10 aprotic solvent, for example, benzene, toluene, ethers, tetrahydrofuran, dimethylsu1 foxide or hexamethy1phosphor-triamide. The reaction temperature may vary from about -120°C-to about 65°C, a preferred reaction temperature being about 40°C. The reaction time will vary from about 15 1/2 hour to 24 hours.

Acid hydrolysis to remove any unreacted starting material and protecting groups may be achieved in one step or stepwise. in a one-step hydrolysis procedure the concentration of acid employed will obviously vary with the 20 duration of the hydrolysis step and temperature employed.

For example, one-step hydrolysis may be achieved by treatment with concentrated hydrochloric acid for 1 to 4 days at about 25° to 120°C. Stepwise hydrolysis may be achieved by treatment with dilute acid for about 1/2 hour to 6 hours 25 at about 25°C to remove unreacted starting material repeating treatment with dilute acid to remove any amine protecting groups followed by treatment with concentrated acid for about 1 to 5 days at about 25° to 125°C to remove any ester or ether groups. Stepwise hydrolysis is preferred. 1 8753S -H+-865' The compounds of general Formula I wherein Ri is ©j\j hydrogen, R2 is hydroxy and cithe-p of R3, R4, R'4 or R5 iiSi i.v- is ORio and Rio is hydrogen are prepared from the corres- A n..^^ ponding derivative wherein either of R3, R4, R'4 or Rs is ORio and Ri0 is methyl by treatment of said derivative with hydrogen bromide in water or acetic acid at a temperature of about 25° to 125°C for about 4 to 24 hours.

Compounds of general Formula 1 wherein Ri is hydrogen, R2 is hydroxy and any of R3, R4, R'4 or R5 is ORio and Ri0 is a straight or branched alkyl group of from 1 to 8 car- bon atoms may be prepared by alkylating the corresponding compound*^where in Ri0 is hydrogen with an alkyl halide of the formula R2iY2 wherein R2i is a straight or branched alkyl group of from 1 to 8 carbon atoms and Y2 is halogen, np U ' Si -fror*- I Jto £ C&J~\oor> exfolr* iU 1^ for example, bromine or iodine^ in a lower alcoholic solvent^ such as methanol or ethanol or hydrocarbon solvents such as benzene or toluene in the presence of an organic base such as triethylamine or pyridine or in an aprotic solvent such as dimethy1formamide, dimethy1acetamide or dimethyl- sulfoxide in the presence of sodium hydride for about 1 to 24 hours at a temperature of about 25°C to 85°C followed by hydrolysis with aqueous base with the proviso that prior to the alkylation reaction the a-amino group and optionally the carboxylic group of the hydroxy substituted ^25 starting material are protected with a suitable protecting group respectively such as tert-butoxycarbonyl or benzyl- oxycarbonyl and benzyl which are subsequently removed by hydrogenolysis or by treatment with acid, such as tri- fluoroacetic acid. The alkyl halides employed in this procedure are known in the art or may be prepared by pro- J4+-885' cedures well known in the art.

The compounds of general Formula I wherein R2 is hydroxy, Ri is hydrogen and any of R3, R4, R'4 or R5 is ORio and Ri0 is alkylcarbonyl wherein the alkyl moiety has from 1 to 6 carbon atoms and is straight or branched, benzoyl, or phenylalkylenecarbony1 wherein the alkylene "moiety is straight or branched and has from 1 to 6 carbon atoms^are prepared by treating the corresponding deriva-tivefl wherein Ri0 is hydrogen^with an acid anhydride of the 0 formula (R22-C-)20 or an acid halide of the formula 0 II R22-C-halo wherein halo is chlorine or bromine and R22 is a straight or branched alkyl group of from 1 to 6 carbon atoms, phenyl or phenylalkylene wherein the alkylene moiety is straight or branched and has from 1 to 6 carbon atoms^ in the presence of an organic base such as pyridine, quino-1ine or triethylamine, which base serves as the solvent, for about 1 to 24 hours at a temperature of about 25°C to 100°C with the proviso that prior to the reaction^the a-amino group and optionally the carboxylic group of the hydroxy substituted start ing mater ial are protected with a suitable blocking group respectively, such as, tert- butoxycarbony1 or benzyloxycarbony1 and benzyl which are / subsequently removed by treatment with acid, forexample, trifluoroacetic acid and hydrogenolysis. The acid anhydride and acid halide reactants employed in this procedure are known in the art or can be prepared from the appropriate acids by procedures well known in the art.

The compounds of general Formula I wherein R2 is a • 1 87 straight or branched alkoxy group of from 1 to 8 carbon atoms are prepared by treating the cor responding derivative# wherein R2 is hydroxy with thionyl chloride to form j, p. &s. the acid chloride which is reacted with an alcohol of the ^ 5.. formula R23-0H, wherein R23 is a straight or branched alkyl group of from 1 to 8 carbon atoms, such as, methyl, ethyl, ji-propyl, isopropyl, ji-butyl, hexyl, or octyl, at about 25°C for from about 4 to 12 hours.

The compounds of general F.ormula 1 wherein R2 is ^0 -NRyRa wherein each of R7 and Re is hydrogen or a straight or branched lower alkyl of 1 to 4 carbon atoms^are prepared cuvadcchori by an acylation' reaction of an acid halide, for example, an . p acid chloride, of the corresponding compound wherein R2 is O I . o o ' " hydroxy and Ri has the meaning defined in Formula I with the proviso that any free amino group is protected with a suitable protecting group, for example, carbobenzyloxy or tert-butoxyearbony 1 and when any of R3, R4, Rs or R'4 is ORio and Rio is hydrogen^said groups are protected as the corresponding a 1kylcarbonyloxy group, with an excess of an OJAU\o doiv^>ou-/\A appropr iateiamino. which may be represented as HNR7Ra» The # reaction is carried out in methylene chloride, chloroform, dimethy1formamide, ethers such as tetrahydrofuran or dioxane ).?. or benzene at about 25°C for about 1 to 4 hours. Suitable gamine3 are, for example, ammonia, or a compound which is a 25 potential source of ammonia, for example, hexamethylene- tetramine; primary amines, for example, methylamine, ethyl-amine, or ri-propyl amine; and secondary amines such as dimethylamine, diethylamine or d i-jn-butylami ne. Following thelacylaticw» reaction the amino protecting group is removed 30 by treatment with acid, for example, hydrogen bromide in ■ 1 875 - ■* dioxane or hydrogeno1ysis, and the hydroxy protecting group, when appropriate, is removed by base or acid hydrolysis.

The compounds of general Formula I.where in R2 is -NH-CH-COOH are prepared by reacting the corresponding Rs derivative wherein R2 is hydroxy^or a functional derivative fp gtS.thereof such as an acid anhydridemand Ri has the meaning V' defined in Formula I with the proviso that any free amino I1 group is protected with a suitable blocking group, such as benzyloxycarbony1 or tert-butoxycarbonyl^with a compound of *0 the formula NH2-CH-C00R24 wherein R9 has the meaning de- ^9 1 So C£xS\& o<\ fined in general Formula I and R24 is a lower alkyl grou^, . o_ for example, methyl or ethyl in an ether, such as, tetra--ip u '<^\....h,ydrofuran or dioxane at 0° to about 50°C for about 1 to 24 hours followed by acid hydrolysis to remove the pro-15 tecting group, 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 compounds of general Formula I wherein Ri is alkylcarbonyl wherein the alkyl moiety is straight or 20 branched and has from 1 to 4 carbon atoms are prepared by treating the corresponding derivatives wherein Ri is hydrogen arid R2 is hydroxy^with an acid halide of the , S- II ^ formula R25-C-halo wherein halo is a halogen atom, for a ^"'"example, chlorine or bromine and R25 is a straight or x y?:-' branched alkyl group having from 1 to 4 carbon atoms^ in water in the presence of a base such as sodium hydroxide ,144-685" or sodium borate at a temperature of from 0°C to 25°C for from 1/2 hour to 6 hours. These compounds may also be prepared from the ester derivative, that is, compounds of general Formula I wherein Ri is hydrogen and R2 is an alkoxy group of from 1 to 8 carbon atoms^by treatment with J* 0 II the acid halide, R25-C-halo, described above, in water, methylene chloride, chloroform or dimethyl acetamide in the presence of a base such as sodium hydroxide, potassium hydroxide or excess triethylamine at a temperature of from about 0°C to 25°C for from about 1/2 hour to 24 hours.

The compounds of general—Formu 1 a—I wherein Ri—Ks ^ a 1koxycarbony1 wherein the alkoxy moiety is straight or branched and has from 1 to 4 carbon atoms are prepared by -treating the corresponding derivative where>ffRi is hydrogen and R2 is hydroxy with an alkyl ha^dformate of the 0 " formula halo-C-0R26 wherein,^halo is a halogen atom such as chlorine or bromine^dri<j R26 is a straight or branched ..a'f kyl group havmg^rrom 1 to 4 carbon atoms in water in the presenc^of a base such as sodium hydroxide or sodium borate^t a temperature of from about 0°C to 25°C for from about 1/2 hour to 6 how^s-* The compounds of general Formula I wherein Ri is 0 « -C-CH-R27 wherein R27 is hydrogen, a straight or branched NH2 lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzyl are prepared by treating the corresponding derivative wherein Ri is hydrogen and R2 is a straight or r:* v.; JiU—665" branched alkoxy group of from 1 to 8 carbon atoms?with an acid of the formula H00C-CH-R27 or an anhydride thereof, c NH2 . \ ?> -wherein the amino group is protected with a suitable -* blocking group such as benzyloxycarbonyl or tert-butoxy-5 carbonyl and R27 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 when the free acid is employed, at a temperature of from about 0°C to 35°C for about 1 to 12 hours^fo1 lowed by *0 acid and base hydrolysis to remove the protecting groups.

Optionally base hydrolysis may be eliminated to give the ester peptide derivative.

The individual optical isomers of the compounds of n i^^genera 1 Formula I wherein Ri is H and R2 is OH may be sepa- \ rated by using a (+) or (-) binaphthylphosphoric acid salt by the method of R. Viterbo et al., Tetrahedron Letters 48, 4617 (197l)» Other resolving agents such as (+) camphor-10-sulfonic acid may also be employed. The individual optical isomers of compounds of Formula 1 wherein Ri and R2 ®feo are other than H and OH respectively may be obtained as described herein for the racemate only starting with the resolved amino acid.

•The compounds of general Formula V may be prepared by ftvoUu" rtreating one/equivalent of a glycinate of the formula \ &"'v" k. ch2-cor_ •" I 9ff.£3 NH=C-Rk Formula VI Rc wherein Rg, R^ and Rc have the meanings defined in general Formula V with one^equivalent of a strong base, such as, ■M4-885" Ci,/\y I alkyl lithium, for example, butyl lithium or phenyl lith- o ium,|lithium di-a1kylamide, for example, lithium diiso-propylamide, or lithium amide, tertiary potassium butylate, sodium amide, metal hydrides such as sodium hydride, tertiary amines such as triethylamine, lithium acetylide^ or dilithium acetylide,followed by treatment with an alkylating reagent of the following formula Ri2 Ri1 R i a CHgXa Formula VII R' 1 2 R1 4 wherein Rn, Ri2, Ri3, R'i2 and Ri4 have the meanings defined in Formula V and X_ is a halogen atom, for example, chlorine or bromine. The alkylation reagent may be carried out in an aprotic solvent, for example, benzene, ethers, tetrahydrofuran, dimethy1su1 foxide or hexamethy1phosphor-triamide. The reaction time varies from about 1/2 hour to 24 hours and the temperature varies from about -120° to 25°C.

The compounds of general Formula VI are prepared by treating an appropriate alkyl glycinate with a carbonyl bearing compound to form a Schiff's base in a generally known manner, specifically, (a) when R^ is hydrogen, by treating the appropriate amino acid ester with benzalde-hyde or an alkanal having from 1 to 9 carbon atoms being straight or branched, for example, 1-propanal, 1-butanal, 2,2-dimethylpropan-l-a1 or 2,2-diethylbutan-l-a1, (b) when R^ 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 7 o (4+^885"" branched, for example, phenyl methyl ketone, phenyl ethyl ketone, phenyl isopropyl ketone, phenyl jn-butyl ketone or phenyl tert-buty1 ketone, and (c) when R^ is a straight or branched alkyl group having from 1 to 8 carbon atoms, treat- ing the appropriate amino acid ester with a-phenyl alkyl o<~ ketone as described above w+-th- a di-alkyl ketone wherein •each alkyl moiety has from 1 to 8 carbon atoms and is straight or branched, for example, dimethyl isopropyl ke-. Y\ . .i-^tbne, di-_n-butyl ketone or methyl tert-butyl ketone. The ■ c carbonyl bearing compounds are "known in the art or may be prepared by procedures well known in the art.

When R. is methoxy or ethoxy in compounds of Formula VI an appropriate alkyl glycinate is reacted with^benzoyl halide, for example, chloride or an alkanoic acid halide, 15 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, tert-butyry1 chloride, 2,2-diethylbutyric acid chloride or valeryl chloride at 0°C in ethers, methylenechloride, di-20 methylformamide, dimethylacetamide, or chlorobenzene in the presence of an organic base such as triethylamine or pyridinej,after which the reaction mixture is allowed to warm to about 25°C for 1 hour. The resulting amide derivative is combined with an alkylating reagent, such as, 25 methylfluorosulfonate, dimethylsulfate, methyliodide, methyl p-toluenesulfonate or trimethyloxonium hexafluoro-'X ^ phosphate when R^ is methoxy or triethyloxonium tetra- fluoroborate when R^ is ethoxy^at about 25°C in a chlorinated hydrocarbon solvent, such as, methylene chloride, 30 chlorobenzene or chloroform, and the reaction mixture is . • t 8 7~ ' * jit-es? '' refluxed for about 12 to 20 hours. The mixture is then cooled to about 25cC and an organic base, such as, tri-ethylamine or pyridine is added after which the solution is extracted with brine and the product isolated.

When in the compounds of Formula VI R^. and Rc to gether form an alkylene group of from 5 to 7 carbon atoms^ * 8( S. said amino acid ester derivatives are obtained by treat- oi ing the amino acid ester with a cyclic alkanone selected from cyclopentanone, eyelohexanone and eyeloheptanone to 10 form a Schiff's base by procedures generally known in t he art.

The alkyl glycinates are obtained by treating glycine with an alcohol of the formula R H wherein R„ has the a a meaning defined in Formula V1^saturated with HC1 gas at 15 about 25°C for about 12 to J>6 hours, or the compounds may be obtained from commercial sources.

The compounds of general Formula VII are known in the art or may be prepared from the corresponding appropriateIy substituted benzoic acid or benzaldehyde derivatives which 20 are known in the art. For example, the benzylhalides of Formula VII may be prepared from the corresponding benzalde- \#&S' J- hyde by reduction with sodium borohydride, lithium alumi- nurn hydride or by catalytic reduction^or from the corresponding benzoic acid ester by reduction.with lithium ^^5 aluminum hydride or boran&jor reduction of the corresponding benzoic acid derivative with lithium hydride and treating the thus formed benzyl alcohol derivative with, for example, thionyl chloride, phosphorus oxychloride, phosphorus trichloride, phosphorus tribromide or phos-30 phorus pentachloride. :?r ^ v. V -M-+—685~~ Certain compounds of general Formula V may be obtained directly from the appropriate amino acid by converting A. J. P.&Ssaid acids to an appropriate ester^then protecting the . u-■§)•..&".3m'no group by the general procedures described herein- above for protecting the alkyl glycinate amino group.

For example, the amino protected ester derivative of tyro- sine or phenylalanine may be obtained in such a manner.

This procedure cannot be employed to prepare compounds of Formula V wherein the aromatic ring contains a hydroxy group in the meta-position however.

The following Example 1 illustrates the use of a compound of general Formula 1 wherein R2 is hydroxy as a chemical intermediate in the preparation of a cephalosporin of Formula I I. j |158cS. EXAMPLE 1 g, 7r T f 2-0 i f1uoromethy1-2-amino-3-phenv1 prop iony1]am ino ]-3- .3.9 " acet y loxymethy 1 -0-oxo-5 - th ia-l-azabicyclo[ 4.2.6 Joet-2^ ene-2-carboxy I ic ac i"d ' A mixture of 1 g of 3-acetylox^-7-amino-8-oxo-5-thia- 1-azabicyclo[4.2.0]oct-2-ene-2-carboxy1ic acid and 1 g of 2-difluoromethyl-2-amino-3-pheny1propionic acid chloride ^^0 wherein the free amino group is protected with tert-butoxy- carbonyl in 50 ml of ethyl acetate is refluxed for 2 hours ,, r after which the solvent is removed leaving a residue which i. ?. Su.cl-v c^- I'NjjdJ'ogeri broi*\»<jle in. or (Xx-e5\c 1 is treated with mild acidjfand chromatographed on silica gel using benzene-acetone as the eluant to give 7-[[2- 3#- d i fluoromethy1-2-ami no-3-pheny1 prop iony1]am i no]-3-acetyloxymethy 1-8-0x0-5-th ia-1-azab icyclo[4.2.0)oct-2-ene-2-carbox-y1ic ac id.

The following Examples 2 to 4 are illustrative of pharmaceutical preparations of the compounds of this 30 invention.

M-K885" EXAMPLE 2 An illustrative composition for hard gelatin capsules is as follows: (a) 2-difluoromethy1-2-amino-3-(3- 20 mg hydroxypheny1)propionic acid (b) taIc 5 mg (c) lactose 90 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 illustrative composition for tablets is as follows: (a) 2-difluoromethy1-2-amino-3-(3,4- 20 mg dihydroxypheny1)propionic 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 injectable suspension is the following 1 ml ampul for an intramuscular injection.

Weight per cent (a) 2-difluoromethy1-2-amino-3-(4- 1.0 hydroxyphenyl)propionic acid (b) polyvinylpyrrolidone 0.5 p ^ rg ^ "^"L' ' '■ J* (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 20 minutes at 121°C. Each ampul contains 10 mg per ml of novel compound (a).

The following Examples further illustrate the compounds of general Formula 1 -a EXAMPLE 5 g^TrrfnTr-^drfiuoTame^try'H^^^T^-d-rtiYaraxYiTh-e n y 1 j propi S -"-afEFiTh" To a stirred solution of 112 g of potassium carbonate in 100 ml of water cooled in an ice-salt freezing mixture is added 25 g of glycine methylester hydrochloride after which the mixture is immediately extracted five times with 15 ether. The combined ether extracts are dried over magnesium sulfate and evaporated at atmospheric temperature to obtain glycine methylester free base. To a stirred solution of 17«7 g of the glycine methylester free base in 150 ml of methylene chloride cooled in an ice bath is ^20 added dropwise 21.1 g (1 equivalent) of benzaldehyde.

After the addition the ice bath is removed and the mixture stirred for 3 hours at room temperature. The methylene chloride phase is washed with brine and aqueous sodium bicarbonate then dried over magnesium sulfate and evap-25 orated under reduced pressure. The resulting residue is distilled at 0.3-0.5 mmHg to obtain the glycine methylester-benzaldimine, b.p. 104°C.

EXAMPLE 6 3,4-D imethoxybenzylbromide To a stirred solution of 50.3 9 of 3>4-dimethoxybenzy1 alcohol in 500 ml of anhydrous ether is added slowly 27.1 g (9.5 ml) of phosphorus tribromide at 0°C. Upon completion of the addition the mixture is stirred at room temperature for 1 hour then poured into a solution of 56 g of potassium carbonate in 50 ml of water. The ether phase is separated, dried over magnesium sulfate and evaporated to give 3,4-dimethoxybenzy1bromide which is recrysta11ized from petroleum ether/b.p. 35-6o°C.

EXAMPLE 7 2-Am ino-2-d i f1uoromethy1-3-(3,4-di hydroxypheny1)propion ic ac i d (A) To diisopropylamine (53 ml, 1 N solution in tetrahydrofuran) is added 25 ml (2.05 M) of n-butyl1ithium under nitrogen with stirring at -78°C. After several minutes the mixture is treated with a solution of the benzaldimine of glycine methylester (8.85 g, 50 mmoles) in 50 ml of anhydrous tetrahydrofuran. After several more minutes a solution of 11.52 g (50 mmoles) of 3*4-dimethoxybenzylbromide in 80 ml of anhydrous tetrahydrofuran is added very slowly during 1 hour with the temperature maintained at -78°C after which the cooling bath is removed and the stirred mixture allowed to return to room temperature during 2-1/2 hours. The mixture is then cooled again to -78°C and treated with a solution of lithium diisopropyl-amide, prepared from diisopropylamine (62 ml, 1 M solution in tetrahydrofuran) and ji-butyl 1 i th ium (27.2 ml, 2.05 M) at -78°C after which the cooling bath is removed and re-38- 1 875' 3 8 44+-S85* placed by a warm water bath at 45° to 50°C. When the temperature of the reaction mixture reaches 45° to 50°C a rapid stream of chlorodifluoromethane is bubbled through the stirred solution for 1 hour with stirring. The cooled mixture is diluted with ether, washed 3 times with brine, dried over magnesium sulfate and evaporated to give crude a-d i f 1 uoromethy 1.^3, 4-d i methoxy pheny ])a 1 an i ne-N-benza 1 d im i ne methylester.

(B) The above obtained crude a-difluoromethy 1^3^4- fecS. .8V- d imethoxypheny l)a lanine-N-benzaldimine methylester (IT.5 g) is stirred with a mixture of 100 ml of ether and 250 ml of N-hydrochloric acid for 6 hours at room temperature. The ether phase is separated off. The aqueous phase is extracted 5 times with methylenechloride then evaporated 15 to dryness under reduced pressure. The residue is refluxed with concentrated hydrobromic acid (250 ml, 47$) overnight under nitrogen after which the solution is evaporated to dryness and the residue treated successively with water and isopropanol (2X), the solutions being evaporated to 20 dryness after each treatment. A nitrogen atmosphere is maintained during these and the following treatments to avoid any oxidation of the catechol. A solution of the residue in 250 ml of water is decolorized with acid washed charcoal, filtered and concentrated to about 75 ml. The 25 pH of the solution is adjusted to 4.5-4.8 using triethyl-amine after which 300 ml of acetone is added. The resulting precipitate is collected, washed with chloroform and recrystal1ized from water/ethano1 (1:5) to give 2-amino-2-d i fluoromethy1-3~(3> 4-di hydroxypheny1)propion ic ac id, 30 m.p. >250°C.

^ MI -88bT example 8 Methy 1 2-benza1d imi ne-3~(4-hydroxypheny1)propionate To an ice cooled suspension of 11.58 g (50 mmole) of tyrosine methylester hydrochloride in 300 ml of methylene-5 chloride is added 5.3 g (50 mmole) of benzaldehyde followed by the dropwise addition of a solution of 5-05 g (50 mmole) of triethylamine in methylenechloride. The reaction mixture is stirred at room temperature overnight then concentrated under reduced pressure to yield a residue which ■et^er IS uxxs>(%ecl ir\ ujaTef~ <Xr\d cvvd is extracted with ether, j Usual work up and Crystal 1 izat ion c S b .&• ' from methylenechloride/pentane gives methyl-2-benza1dimine- !?-(4-hydroxyphenyl ) propionate, m.p. 109-H1°C.

EXAMPLE- - 9 -7'- Methy1 2-chloromethy1-2-benzaIdim ine-5-(4-hydroxyphenyl)-prop ipnate To a stirred solution of lithium diisopropylamide (8 mmole) prepared in situ from a solution of 1 mp4ar ' diisopropylamine in tetrahydrofuran and a solution of 2 molar butyl lithium in hexane, in tetrahydrpfuran and hexa-methylphosphortriamide (2.5 ml) cooled ,to -78°C is added 20 slowly under a nitrogen atmosphere ar solution of 1.132 g (4 mmole) of methyl 2-benza1dimlde-3-(4-hydroxypheny1)-propionate in 30 ml of tetrahydrofuran. The cooled re- r / action mixture is stirre^at 1/2 hour at -78°C after which ^solution of chlorob/'omopropane (1.05 g, 8 mmole) in 25 tetrahydrofuran i^added. The temperature is allowed to rise slowly to/room temperature and stirring is continued overnighty/The reaction mixture is quenched with water and ex£/acted with ether. Usual work-up and crystallization from ether/pentane gives methyl 2-chloromethyl-2- n • a 17 ^ \js Ml-885 bengal d imina-3-( i|-hydro* yphonyl )propionate^—m<p. <50°C.

EXAMPLE 10 2-Am ino-2-ch loromethy) -3- ( 4-hydroxy phenyl) prop ion ic ac id (A) A solution of 645 mg of methyl 2-chlorometh' 5 2-benzaIdimine-3-(4-hydroxyphenyl)propionate in 10/ml of ether is virogously stirred with 10 ml of 1 nonrfal hydrops- chloric acid for 2-1/2 hours at room temperature then II ' eft, Extracted with ether. The aqueous phase/is concentrated vv-"-"'' / under reduced pressure to give methyl/2-amino-2-chloro-10 methyl-J-(4-hydroxyphenyl)propionate as an oily residue. ^ (B) A solution of 450 mg/(l.6 mmole) of methyl 2- am ino-2-chloromethy1-3-(4-hydroxypheny1)prop ionate hydro-chloride in 10 ml of consent rated hydrochloric acid is refluxed for 15 hours^f The resulting brown solid obtained 15 upon concentrat iocr under reduced pressure is taken up in dilute hydrochJxjric acid and decolorized with charcoal after which/the solvent is evaporated and the residue dried under vacuum to give a white powder which is washed twice with/water yielding 2-amino-2-ch1oromethy1-3-(4-hydroxy-20 -phreny 1 )propionic—ac i d j—m. p.—£56° C .

EXAMPLE yS°[ p.O&pionate hydrochloride | Q. S'.

'?• Ethyl 2-difluo romethy1-2-am i no-3-(3,4-d ihydroxyphenyl) ■ A suspension of 2.47 g (10 mM) of 2-difluoromethyl-2-amino-3-(3j4-dihydroxypheny1)prop ionic acid in 30 ml of ^5 ethanol is saturated with anhydrous HC1 and the resulting solution allowed to stand at 25°C for 24 hours. The solvent is evaporated leaving a residue which is recrysta11ized from ethanol-ether to give ethyl 2-difluoromethyl-2-amino-3-(3> 4-d ihydroxyphenyl)prop ionate hydrochlor i de. f T, / ^ e> EXAMPLE ¥2'la 2-D i fluoromethyl-3-(3,, 4-diacetyloxyphenyl)-2-(benzyloxy-carbonyl am i no) prop i on ic acid 2 N aqueous sodium hydroxide and acetic anhydride . ^ N. *• ' o, (3„.5 g) are added simultaneously during 1/2 hour to a solu--.l-Vyl1'*' - v, 5'"' tion of 2-d i f 1 uoromethy 1 -2- (benzy loxycarbony 1 am i no)-J- (3,4-dihydroxyphenyl)propionic acid (6 g ) prepared from 2-difluoromethy1-2-amino-3-(3>4-dihydroxypheny1)propionic acid and benzyIchloroformate, in 30 ml of 1 M sodium hydroxide under argon so that the pH is maintained between 6.5 and ^10 7-5- After 1 hour at 25°C the pH is adjusted to 1 using 6 N sulfuric acid and then the solution is extracted with methylenechloride. The organic phase is dried and concentrated to give 2-difluoromethyl-2-(3*4-diacetyloxypheny1)-2-(benzy1 oxyearbony lam ino)prop ion ic ac id. 15 EXAMPLE \ 2-D i f1uoromethy1-2-(acetyl am i no)-3-(3,4-d i hydroxypheny1)-o. * propionic acicT , , To a stirred solution of 6.8 g of borax in 10 ml of water is added 2.47 g (10 mM) of 2-dif 1uoromethyl-2-amino- 3-(3j4-dihydroxyphenyl)propionic acid under argon. After ^^0 15 minutes the pH is adjusted to 9 by the addition of 2 N sodium hydroxide then j80 mg of acety1ch1 oride is added dropwise while the pH is maintained between 9*0 and 9-5. The aqueous solution is washed with ether, adjusted to a % pH of 1 using 6 N sul furic acid and extracted with methylene- chloride. The organic phase is dried and concentrated to afford 2-d i f1uoromethy1-2-(acetyl am i no)-3-(3> 4-d ihydroxy-pheny 1) prop ion ic acid which may be treated with IC A HC1 to afford the corresponding methylester.

EXAMPLE 2-r2-D ifluoromethy1-2-amj no-3-(3> 4-diacetyloxyphenyl)-1-oxo-propy lami no | prop i on i c acid A solution of 4.66 g (10 mM) of 2-difluoromethy1-2-3c, d.U-'^'"'Ccarbobenzyloxyamino)-3- {?>> 4-d iacety loxyphenyl) propion ic 5 acid, prepared from 2-difluoromethy1-2-amino-3-(3*4-diace-tyloxyphenyl)prop ionic acid and benzylchloroformate, in 50 ml of ether is treated with 1.0 g (10 mM) of triethyl-amine followed by 1.08 g (10 mM) of ethylchloroformate.

After 1 hour at 25°C the precipitate is filtered off and 10 to the ether solution is added a solution of alaninebenzyl-ester (10 mM) in 30 ml of ether. The solution is maintained at 25°C overnight then evaporated to dryness„ The residue is treated with HBr in dioxane (40^ w/w, 20 ml) for 30 minutes at 25°C. Ether is then added and the precipitated 15 hydrobromide filtered off to give 2-[2-difluoromethy1-2- am i no-3-(3i4-diacetyloxyphenyl)-1-oxopropylam i no]prop ion ic acid.

EXAMPLE y5\Z \ ? & S K. J- 2-D i fluoromethy1-2-(2-am i no-1-oxopropy1 am ino)-3-(3? 4- t\ ) pre ' ' x ^ ,g.l..-d'f hydroxypheny I ) prop i on ic acid hydrochloride A suspension of 3-56 g (10 mM) of benzyl 2-difluoro- methyl-2-amino-3-(3j4-dihydroxypheny1)propionate in 50 ml of methylenechloride is treated with 1 g (10 mM) of tri-ethylamine after which 10 mM of N-carbobenzyloxyaIanine wherein the acid function is activated by ethoxycarbonyl 25 in 10 ml of methylenechloride is added. The mixture is stirred at 25°C for about 16 hours then washed with water. The organic layer is dried and evaporated. The residue is taken up in ether and the ether solution cooled to 0°C. A vigorous stream of HC1 gas is bubbled through the solu- • 1875 3 -H+^885 tion for 3 hours after which the ether solution is washed with water. The aqueous phase is evaporated to afford 2-d i f 1 uoromethy 1 -2- (2-ami no- 1-oxopropy 1 ami no)-J>- (3, 4-d i -hydroxypheny1)prop ionic acid hydrochloride as a gum. . example a&m- 1 P. , 2-D i fluoromethy1-2-carbobenzoxyam i no-3-(3> 4-di hydroxypheny1)- M 9l—.^opion ic acid To a solution of 8.2 g of 2-difluoromethy1-2-amino-3-(•3, 4-d i hydroxypheny 1 ) propion ic acid and 12.7 g of borax in 80 ml of water cooled to 0°C and kept under nitrogen is ^10 added dropwise 7-5 g of benzylchloroformate over a 1 hour period, the pH of the solution being held to 9 by the addition of 2 N aqueous sodium hydroxide. Stirring is continued for 1 hour after the addition and the undissolved material is filtered off. The filtrate is extracted several times with ether. The organic phases are discarded and the pH of the aqueous phase is adjusted to 1 by addition of concentrated sulfuric acid with cooling. Extraction with ether yields crude 2-difluoromethy1-2-carbobenzoxyam i no-3- (3 j 4-d i hydroxypheny 1 )propionic acid.

When in the above procedure an appropriate amount of ^ acetylchloride or benzoylchloride is substituted for ben zylchloroformate, 2-d i f1uoromethyl-2-acetyl ami no-3-(3> 4-dihydroxyphenyl)propionic acid and 2-dif1uoromethy1-2-benzoylamino-3-(3j4-dihydroxypheny1)prop ionic acid are 25_ obtained respectively. » & example yr l< g J _ 2- (2-Amino-2-d i f 1 uoromethyl -3- pheny 1 - l-oxopropaneamino)-propion ic ac id £ To a solution of 3.15 g of N-tert-butoxycarbonyl-g- difluoromethylphenylalanine in 30 ml of ether is added 1 87 r • <L-.v C"' -M+*685~ 1.01 g of triethylamine followed by an ethanol solution of freshly distilled ethylchloroformate (1 g). The reaction mixture is stirred for 3 hours at room temperature and the precipitated triethylamine hydrochloride filtered off.

To the filtrate is added at once a solution of 1.4-5 g of alanine tert-bgtylester in ether. The reaction mixture is maintained overnight at room temperature and then evaporated to dryness. The residue is taken up in tri-fluoroacetic acid. The solution is concentrated and the 10 residue purified by ion exchange chromatography on a resin to give 2- (2-ami no-2-d i fl uoromethy 1 -3-phenyl - l-oxo^propane-amino)propionic acid. alanine methylester in 30 ml of ether cooled to 0°C is added under a nitrogen atmosphere a solution of N-benzyl-oxycarbony1-0-ethoxycarbony1 a 1 anine (2.9 g) in ether.

Upon completion of the addition the cooling bath is re-20 moved and stirring continued overnight. The solution is evaporated and the syrupy residue taken up in 5 ml of methanol and 10 ml of 2 N aqueous ammonia. The reaction mixture is stirred overnight at room temperature then extracted with ether several times. The aqueous phase is 25 neutralized and 2-(2-amino-l-oxopropylamino)-2-difluoromethyl -3-pheny1 prop ionic acid is isolated by ion exchange chromatography on an Amber lite IR 120 H+ resin.

EXAMPLE I \o , n i.^••'2- (2-Am i no-l-oxopropy 1 am i no) - 2-d i fl uoromethy 1 -3- pheny 1 -propionic acicf To a solution of 2.3 g of a-d i fl uoromethy 1/phenyl-

Claims (13)

w t;-'' EXAMPLE l~7 Methyl 2-difluoromethyl-2-amino-3-(3,4-d imethoxypheny1)-propionate k * To a solution of methyl*2-d if luoromethyl-2-benza Id- Q\ •^.'••■'•rrfime-3-(3> 4-d imethoxypheny 1) propionate (30 g) in ether 5 (200 ml) is added a solution of HC1 0.5 M (15 ml). The heterogenous mixture is stirred at room temperature for 12 hours. The organic phase is separated and then a solution of HC1 M (200 ml) is added. After stirring the reaction mixture 1 hour at roorp temperature the aqueous ^^10 phase is separated and concentrated under reduced pressure to give methyl }f2-d i fl uoromethyl-2-ami no-2-(3* 4-d imethoxy-phenyl)propionate hydrochloride. EXAMPLE J2Xf | ST \ P. & ^h-D i f1uoromethyl-2-am i no-3-(3>4-d ihydroxyphenyl)prop ion ic - ac id A solution of methyl 2-dif1uoromethyl-2-amino-3-(3j4-dimethoxypheny1)propionate hydrochloride (20 g) in 47$ aqueous HBr (200 ml) is heated under N2 at reflux temperature for 12 hours. The residue obtained after concentration under reduced pressure is dissolved in water (30 ml) and ^20 the pH of the solution is adjusted to 4.5 with a solution of NEt3 and acetone is added until precipitation starts. 2-D i fluoromethy1-2-ami no-3-(3j 4-d ihydroxyphenyl)propion ic acid dihydrate is collected by filtration, m.p. >26o°C. -46- 187538 WHAT WE CLAIM is: 1. A compound of the formula wherein Y is FCH2 or F2CH-; R^ is hydrogen, alkylcarbonyl wherein the alkyl moiety has frcm 1 to 4 carbon atoms and 0 ll is straight or branched or -C-CH-R27 wherein R27 is hydro- NH2 gen, a straight or branched lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzy1; R2 is hydroxy, a straight or branched alkoxy group of from 1 to 8 carbon atoms, -NR7R8 wherein each of R7 and R8 is hydrogen or a straight or branched alkyl group of from 1 to 4 carbon atoms, or -NH-CH-COOH wherein Rg is hydrogen, a straight R 8 or branched lower alkyl group of from 1 to 4 carbon atoms, benzyl or p-hydroxybenzy1; and each of R3, R4, R5, R'4 and R6 has the meaning defined in Table I wherein Rio is hydrogen, alkylcarbonyl wherein the alkyl moiety is straight or branched and has from 1 to 6 carbon atoms, benzoyl or phenyla1kylenecarbonyl wherein the alkylene moiety is straight or branched and has from 1 to 6 carbon atoms; AS AMENDED TABLE 1 £•2. fLi Rs R' 4 Re H -o-ch2 -0- H H H H OR i o H H H OR i o H H H H O tH cc o OR i o H H ORi o H C 1 H H H ORio C 1 H H CI OR i o H H H C 1 ORio CI H H ORio H ch3 H CHa ci H ch3 h ch3 ORi o H c2h5 . H CeHs H ORio H OR i o H H ORio ORio ORio u

1 1 H H och3 OH H OR i o OR i o H H H OR i o H H H H excluding compounds wherein (a) Y-CH^F; , R1^, and R =H; R2=-0H or alkoxy; and and R^ both are -OR-^, wherein s is H or alkylcarbonyl, (b) Y=CH2F; R^, R^, R4, and Rg=H<" ^2 ~0H or ailKOXY R5=-OH, or (c) Y=CH2F; R^, R3, R'^, Rg, and Rg=H; is -OH or alkoxy and R^=-OH, htz t - 48 ,187538 | NOW AMEMDEDI and pharmaceutica11y acceptable salts and individual optical isomers thereof. *

2. A compound of claim 1 wherein Ri/ is hydrogen or alkylcarbonyl wherein the alkyl moiety bias from 1/to U carbon atoms and is straight or branched.

J>. A compound of claim 1 where/in R2 is h/droxy or a straight or branched alkoxy group c/f from 1 t<f 8 carbon atoms.

4. A compound of claim 1 yfiere in ^ 4 '5 hydrogen.

5. A compound of claim 4 yiwherein Ri/is hydrogen and R2 is hydroxy.

6. A compound of claim 1 wherein/Y is FCH2-

7. A compound of claim i where/n Y is FzCH-} R2 is hydroxy and Ri is hydrogen.

8. A compound o/t claim 1 wraich is 2-dif1uoromethyl 2-amino-J-[J>} 4-d ihydi^oxyphenyl) p/opion ic acid or a pharmaceutical ly acceptab/e salt thereof. 9. A compo/ind of claim/l which is 2-dif1uoromethyl-2-amino-3_(3-hydroxyphenyl)p/opionic acid or a pharmaceutical ly acceptable salt /thereof. io. A compound of c/aim 1 which is 2-d i f 1 uoromethyl -QA / / 26\ami no-J-(¥-hydroxyphen»l) prop ion ic acid or a pharmaceu- .* ~4 FEBJ982 ! /■ / t^cally acceptable sa1t/thereof. 11/ A process for preparing a compound of claim 1 which comprises i) when Ri Vs hydrogen, R2 is hydroxy and Rio is .hydrogen by treating a su i t/ab 1 y protected phenylpro- - 49 - "49_ 187533 and pharmaceutica11y acceptab1e sa1ts and individual optical isomers thereof^ ' AS AMENDED 2. A compound of claim 1 wherein Ri is hydrogen or alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and is straight or branched. 3- A compound of claim 1 wherein R2 is hydroxy or a straight or branched alkoxy group of from 1 to 8 carbon atoms. 4. A compound of claim 1 wherein ^10 's hydrogen. 5. A compound of claim 4 wherein Ri is hydrogen and R2 is hydroxy. 6. A compound of claim 1 wherein Y is FCH2- 7. A compound of claim 1 wherein Y is F2CH-j R2 is hydroxy and Rx is hydrogen. 3. A compound of claim 1 which is 2-difluoromethyl-2-amino-3-(3*4-dihydroxypheny1)prop ionic acid or a pharmaceutical ly acceptable salt thereof.

9. A compound of claim 1 which is 2-difIuoromethyl-2-amino-3-(3-hydroxypheny1)propionic acid or a pharmaceutical ly acceptable salt thereof.

10. A compound of claim 1 which is 2-difluoromethyl-2-amino-3-(4-hydroxypheny1)propionic acid or a pharmaceutical^ acceptable salt thereof.

11. A compound as claimed in any one of claims 1 to 10 substantially as hereinbefore described with reference to any of Examples 7 and 9 to 18. - 50 - 1875 pionate of the formula ri2 rii ris-./fy. ch2-ch-c0r [NOW AMENDED I i mil — || , wherein R= is a straight or branched alkoxy group having a -it from 1 to 8 carbon atoms; R^ is hydrogafi, phenyl, i straight or branched alkyl group of from 1 to a carbon atoms, methoxy or ethoxy; Rc is phenyl or a straight or branched alkyl group of from 1 to 8 carbon atoms; or R^ ind Rc toget/her form an alkylene group of from 5 to 7 carbon atoms; and each of Ri 1, Ri2, R13, R'12 and Ri4 has tine meaning ytlefined in the following Table II: TABLE Ri i Rjj lLS. _L£ Ri/ h -0-c-0- /\ h3c ch3 -o-ch£-o- /H h • h h 0ch3 h och3 /h h och3 / och3 ocha H / ci h 0ch3 / ci ci ochl h ci 0chs ci , och: ch3 50 AS AMENDED '87538 12. A process substantially as hereinbefore described with reference to any of Examples 7 and 9 to 18. 13. A compound as claimed in any one of claims 1 to 10 when prepared by a process, substantially as hereinbefore described. dated day Of lOo bilt' A. J. PARK & SON agents for the applicants - 51 - och3 H c2hs H c2h H och3 H och3 H H 0ch3 och3 ocha H H H 0ch3 0CH2Ph H 0CH3 OCHa H H H ocha H H H H with a suitable strong base and treating the thus obtained carbanion intermediate with a suitable halorriethyl alkylating reagent at a temperature of -120°C to 65°C for 1/2 hour to 24 hours in an aprotic solvent followed by (I) acid hyrolysis and, if necessary to cleave the aromatic methyl ether group, treatment with hydrogen bromide' in water or acetic acid for 4 to 24 hours at 25°c to 125°c or (II) treatment with hydrogen bromide in water or acetic cm ...acid for 4 to 24 hours at 25°C to 125°C: (b) when Ri is hydrogen, R2 is hydroxy and any of R3, R4, R5 or R-4 is ORio and Rlo is alkylcarbonyl wherein the alkyl moiety has from 1 to 6 carbon atoms and is straight | or branched, benzoyl or phenylalkylenecarbonyl wherein the alkylene moiety is straight or branched and has from 1 to 6 carbon atoms, treating the corresponding amine and optionally carboxylic protected derivative wherein Rio is 0 hydrogen,with an acid anhydride of the formula (R22~C-)20 0 II or an acid halide of the formula R22-C-halo wherein halo is chlorine or bromine and R22 is a straight or branched alkyl group of from 1 to 6 carbon atoms, phenyl or phenylalkylene wherein the alkylene moiety is straight or branched and has from 1 to 6 carbon atoms, in the presence of an organic - 52 - 18/5 base which serves as the solvent for 1 to 24 hours at 25° to 100°C,. fol lowed by acid hydrolysis or hydrogenolysis; (c) when R2 is a straight or branched alkoxy group of from 1 to 8 carbon atoms, reacting the corresponding derivative wherein R2 is hydroxy with thionyl chloride followed by treatment with an alcohol of the formula R230H wherein R23 is a straight or branched alkyl group of from 1 to 8 carbon atoms at about 25°C for 4 to 12 hours; (d) when Rs is -NR.t-Rs wherein each of R7 and Rs has the meaning defined in claim 1,treating an acid halide of the corresponding derivative wherein R2 is hydroxy and Ri has the meaning defined in claim 1 with the proviso that any free amino group is protected with a suitable protecting group, and when any of R3, R4j Rs or R'4 is 0RiO and Rio is hydrogen, said groups are protected as the corresponding a 1 kylcarbonyloxy group, with an excess of an appropriate amine of the formula NHR7R8 wherein R7 and Rs have the meanings defined above,in an appropriate solvent at about 25°C for 1 to 4 hours, followed by acid or base hydrolysis or hydrogenolysis; (e) when R2 is -NH-CH-C00H wherein Rg has the meaning I Re defined in claim 1, treating the corresponding derivative wherein R2 is hydroxy or a functional derivative thereof and Ri has the meaning defined in claim 1 with the proviso that any free amino group is suitably protected, with a compound of the formula NH2CH-C00R24 wherein Rg has the Rs meaning defined in claim 1 and R24 is a lower alkyl group having from 1 to 8 carbon atoms, - 53 - in a suitable solvent at about 50°C for /I to 24 hours, followed by acid hydrolysis with the proviso that when the amine protected free acid is employed the reaction is carried out in the presence of a dehydrating agent; (f) when Ri is alkylcarbonyl wherein the alkyl moiety has from 1 to 4 carbon atoms and is straight or branched, treating the corresponding derivative wherein Ri is hydrogen, and R2 is hydroxy,with an acid halide of the formula <0 II Rfs-C-halo wherein halo, is a halogen atom and R25 is a straight or branched alkyl group of from 1 to 4 carbon atoms,, in a suitable solvent in the presence of a base at a temperature of from 0°C to 25°C for from j 1/2 hour to 6 hours; / ° (g) when Ri is -C-CH-R27 wherein R27 has the meaning !jh2 defined in claim 1, treating the corresponding derivative wherein Ri is hydrogen and R2 is a straight or branched alkoxy of from 1 to 8 carbon atoms, with an acid of the formula H00C-CH-R27for an anhydride thereof, where in R27 nh2 has the meaning defined above and the amino group is suitably protected, in a suitable solvent and in the presence of a dehydrating agent when the free acid is employed, at a temperature of 0°C to 35°C for 1 to 12 hours, followed by acid and base hydrolysis; and (h) when a pharmaceuticaI 1y acceptable salt is desired, reacting the thus obtained compound with a pharma-ceutically acceptable acid or base.

12. A compound as claimed in any one of claims 1 to 10 I - 54 - 187533 substantially as hereinbefore described with reference to any of Examples 7 and 9 to 18.

13. A process as claimed in claim"11jsubstantially as hereinbefore described with reference to any of Examples 7 and 9 to 18. dated this^K day of ocfis bar t9 <27 A. J. PARK & SON AGENTS FOR THE AOPUOa.s 13 '20 OCT 1981