IE43478B1 - New 16-ethers of fusidic acid derivatives - Google Patents

Abstract

A new series of fusidic acid derivatives, salts and easily hydrolyzable esters thereof, and processes for the preparation of these compounds are provided herein. The new compounds have the general formula in which the C24-25 bond is a single or a double bond, and in which Q1 and Q2 stand for or oxygen; A stands for oxygen, sulphur or a sulfinyl radical; R1 stands for a straight or branched alkyl radical having from 1 to 8 carbon atoms, or such straight or branched alkyl radical having from 1 to 8 carbon atoms when substituted by halogen atoms of hydroxy, alkyloxy, aralkyloxy, aryloxy, alkanoyloxy, aralkanoyloxy, aroyloxy, sulfhydryl, alkylthio, aralkylthio, arylthio, alkanoylthio, aroylthio, azido, nitro, cyano, thiocyano, hydroxycarbonyl, alkyloxycarbonyl, amino, alkylamino, dialkylamino, arylamino, ??anoylamino or aroylamino groups; an alkenyl or alkynyl radical having from 7 to 6 carbon atoms, a cycloalkyl radical having from 3 to 7 carbon atoms in the alicyclic ring, an aryl, aralkyl or heterocyclylalkyl radical, or such alkenyl or alkynyl radical having from 2 to 6 carbon atoms, such cycloalkyl radical having from 3 to 7 carbon atoms in the alicyclic ring, such aryl, such aralkyl or such heterocyclylalkyl radical when substituted by halogen, nitro, lower alkyl, hydroxy or alkoxy radicals: or a heterocyclic radical having 5 or 6 ring atoms, containing oxygen, sulphur or nitrogen atoms, and selected from 2- or 3-pyrrolyl, 2- or 3-furyl, 2- or 3-furfuryl, 2- or 3-thienyl, 2-, 3- or 4-pyridyl, 2-, 4or 5-pyrimidinyl, 2- or 3-pyrazolyl, imidazolyl, 1-methyl-2-immdazolyl, triazolyl, 5-methyl-1,2,4-triazol-3-y1, tetrazoyl, 1-methyl-1H-tetrazol-5-yl, thiazolyl, thiadiazolyl and 5-methyl-1,3,4-thiadiazol-2-yl: and pharmaceutically acceptable salts and easily hydrolyzable esters thereof. These compounds show interesting antimicrobial and pharmotokinetic properties. These compounds can be used in the treatment of bacterial infections in humans and animals.

Description

The present invention relates to a new series of fusidic acid derivatives,, to salts and easily hydrolyzable esters thereof, to the preparation of these compounds, and to pharmaceutical compositions containing the compounds. The new compounds have the general formula in which and each stand for the group jj^>or oxygen, A represents oxygen or sulphur or a sulphinyl radical, and stands for a straight or branched alkyl radical which has from 1 to 8 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert,butyl, the known isomers of'pentyl, hexyl, heptyl and octyl, and which is unsubstituted or substituted with one or more halogen _ atom» or hydroxy, alkyloxy, aralkyloxy, aryloxy, alkanoyloxy, oralkanoyloxy, aroyloxy, sulfhydryl, alkylthio, aralkylthio, arylthio, alkanoylthio, aroylthio, azido, nitro, cyano, thiocyanato, hydroxycarbony.1, alkyloxycarbonyl, aryloxycarbonyl, amino, alkylamino, dialkylamino, arylami.no, alkanoylamino, and aroylamlno groups; R^ can further be an alkenyl or alkynyl radical having from vtjto 6 carbon atoms, such as allyl, crotyl or propargyl, a cycloalkyl radical having from 3 to 7 carbon atoms in the alicyclic ring, such as cyclopropyl, cyclobutyl, cyclopontyl, cyclohexyl, cyclohoptyl, or a mono- or dihalo, lower alkyl, lower «* alkoxy or hydroxy substituted analogue, thereof an aralkyl, heterocyelylalkyl or aryl radical, such as benzyl, phenylethyl, phenyl or furfuryl, which is unsubstituted or substituted with one or more halogen atoms or nitro, lower alkyl, hydroxy or alkoxy radicals; Rj can also be an unsubstituted of methylsubstituted heterocyclic radical having 5 or 6 ring atoms and containing oxygon, sulphur or nitrogen atoms, such as 2- or 3-pyrrolyl, 2- or 3-furyl, 2- or 3-thienyl, 2-, 3- or 4-pyridyl, 2-, 4- or 5-pyrimidinyl, 1- or 3pyrazolyl, imidazolyl e.g. l-methyl-2-imidazolyl, triazolyl o.g. 5-«iothyl-l,2,4-triazol-3-yl, tetrazolyl o.g. l-mothyl-lH-tetrazol-5-yl, thiazolyl, thiadiazolyl o.g. -methyl-1,3,4-thiadiazol-2-yl.

In formula I the dotted line between C-24 and C-25 indicates that the carbon atoms in question are connected - 3 4S478 with either a double bond or a single bond.

Whei’o no t o therwise stated the term lower” as applied to the i-adicats mentioned above and others mentioned hereinafter stands for a to C radical. ίο Of particular interest are compounds in which and. Qg are both a group, or one of Qj or Qg is oxygen, A represents oxygen or sulphur or a sulphinyl radical, and R^ stands for a straight or'branched alkyl group with from 1 to 4 carbon atoms, optionally substituted with halogen atoms, hydroxy groups, or an azido group, and the bond between 0-24 and G-25 is either a double bond or a single bond.

The compounds which are especially preferred of the just mentioned groups of compounds are those in which is ethyl or isopropyl, optionally substituted with fluorine.

The compounds of the invention can be used as such or in the form of salts or easily hydrolysable esters (as hereinafter defined), The salts of the compounds are especially the pharmaceutically acceptable, non-toxic salts, such as a-lkali metal salts and alkaline earth metal salts, for example sodium, potassium, magnesium or Calcium salts, as well as salts with ammonia or suitable non-toxic amines, such as lower alkyl aminos, for example triethylamine, hydroxy-lower alkylamines, for example 2-hydroxyethylamine, bis-(2-hydroxyethyl}amine or tris-(2-hydroxyethyl)-amino, cyeloalkylaminos, for example dicyclohexylamine, or benzylamines, for - 4 43478 examplt· NjN’-dibenzyl-ethylonediamine, and dihenzylamine.

For certain purposes also tlie silver salts oi' the compounds may ho used, especially for local treatment.

The expression easily hydrolysable esters is used in this Specification to denote alkanoyloxyallcyl, aralkanoyioxyalkvl, aroyloxyalkyl esters, such as acetoxymethyl, pivaloyloxymethyl, benzoyloxymethyl esters, and the corresponding I’-oxyethyl derivatives, or alkoxycarbonyl oxy alkyl esters, such as methoxycarbonyloxymethyl, ethoxycarbonyloxymetiiyl esters, and the corresponding I -oxyethyl derivatives, or lactonyl esters, such as phtlialidyl esters, or dialkylaminoalkyl esters, such as diethylaminoethyl esters.

The antibacterial properties of fusidie acid are well known, and it is also known that variations in the structure may cause a complete loss of such activity.

Now, however, it has been found, that the compounds of the present invention both in vitro and in vivo show interesting antimicrobial and pharmacokinetic propertiesj Whereby the compounds of the invention can be used in the treatment of bacterial infections in humans and animals. In vitro investigations have for instance shown that the compounds are highly potent against a number of bacteria, e.g. staphylococci, streptococci, corynebacteriae, neisseriae, clostridiao and bacteroidos species, and Bacillus sub tillsj as can be seen from the following table: activity of compounds of1 formula ri ri •ri fc Φ P ri ri •P Ό •ri W Ο ri 01 ri in TO TO Ο TO 1-, +i 6a QUO fi fi o TO c in O cn ri . vo • ri 01 cn 2.0 o in 1.0 cn «ri 01 cn o 2.0 CM cn cn vo o ε. w tn to ri Ή ΟΊ zi. TO TO TO CO 01 ri 'ri Ol o O VO Ol m in in in lf> cn oi -t o •ri -P o o -d* •ri o o o o o o o O o Cl β 0 43 0 • • • • • • • • • a 0 ri ri Φ o o o o o o o o o o o o o » o ri TO ® J •ri w oi to • φ ϋ ft-ri ft o Ol o cn σ\ TO Φ O o ri cn m vo r* o o 01 in o o 01 VO is fi 0 0 • • • * • • * • •ri P ft Φ ω w ri ri o o o o o iri in cn ol rri cn ri 1 m 0 ft 0 fi ft Φ 0 TO ft O TO· o vo o 01 o cn O o ft to Ol o oi ri 01 cn -if o o vo vo in m tr- τί φ Φ • • • • * • φ fi fi o o o o . o o o ri iri o •ri o o O Fl ρ φ o ri cn 6a u 3 σ4 0 1 fi ri o ri φ m « • J co m β TO c- 01 vo o o o o o m O o in vo 01 0 ft «I TO cn ri in -if 01 -if rit , 01 01 Ol Ol •ri 01 cn •ri fi 3 o O o O o. o o o o o o o o o •P Ρ Φ o • « • • ri cn fi o o O o o o o o o o o o o o o fc •P β in φ oi Φ Φ Φ Φ Φ Φ Φ © Φ © © Φ © ϋ •k rri •ri ri rri tri rri iri fri iri fi -’t τι TO 60 TO to TO 60 TO TO TO TO 43 TO TO 0 CM fi ri fi ri fi ri ri ri ri ri 3 3 ri 3 Ο I 0 0 o ri 0 •ri 0 o 0 o 0 0 0 a 43 Ό 01 Ό 01 Ό 01 ri t) ri ri ri ri ri 01 ol Ol 01 01 01 01 01 z-x -*x, z—X Z^X z-x z-x Z—X z—x ft cn cn cn cn cn cn cn cn Ol Ol cn oi ft TO B s B B B w w B B B ft B o o a o o o ϋ o a o u ϋ o \_Z x_>» X^Z >««z S^Z 01 Ol Ol Ol Ol X_Z B B B B B B s B B B B B B 0) o u u υ u o Cl o o o o o ϋ ri fi Φ 3 TO CO co o ω o co ω w OT o o o o o o O ri ri P B B S B B B B B B Ό w o o o o o o ϋ o o ri rO 01 1 1 | 1 1 1 1 I 1 ri Qf y y y a o o b y o y o y y 3 w •k •k Zk Zk Z, Zk ·, ft B B B B B B B B B B B B B B B B B B B B W ri σ o o υ a o o o o O o o ϋ y y y y b y o y y y b y y B B B B B B B B B B B X ( D ri ft ε ri ri m 01 01 cn -t VO m o vo CO 01 ri · cn •if cn cn cn 4· m t- -if m M a 63478 ίο IS Furthermore the compounds of the invention are chemically more stable than fusidic acid, for instance tiie l6-acetoxy group in fusidic acid will under certain conditions be hydrolysed to a hydroxy group in which case a considerable decrease of activity takes place. Xn the compounds of tho invention the 16-radicals are not inclined to such hydrolysis, e.g. when 256 (w/v) solutions of the compounds of the invention in aqueous buffer of pH 9.6 were kept at 4o°C for days, not even traces of degradation products could be detected by thin layer chromatography.

Like fusidic acid tho new compounds are absorbed efficiently from the gastro-intestinal tract and are practically non-toxic.

Tho compounds of formula I can be prepared by a method comprising a first step in which a compound of the general formula II is reacted with a compound of the general formula III to form a compound of the general II IV III - 7 43478 in which formulae stands for as dofinod above or for R , R2 representing an alkanoyl, an aralkanbyl or an aroyl radical; Qg and the dotted line between 0-2^1 and C-25 have the meaning as defined above, the wavy line between C-16 and the hydroxyl group indicates that the latter can be α-ox'iented or β-oriented; X stands for hydrogen or a salt-forming cation, such as Na+,K+,Ag+, an ammonium or trialkylammonium ion, Z is a chlorine, bromine or iodine atom, and R^ represents a straight or branched alkyl radical having from 1 to 6 carbon atoms, e.g. * methyl, ethyl, tert, butyl, an unsubstituted or substituted aralkyl radical, e.g. benzyl, p-nitrobenzyl, or p-inethoxybenzyl, an alkanoylmethyl or aroylmethyl radical, e.g. acetonyl or phenacyl, an alkanoyloxyalkyl or aroyloxyalkyl radical, e.g. acetoxymethyl, pivaloyloxymethyl or benzoyloxymethyl, an alkyloxymethyl radical or a cyanomethyl radical.

The reaction is performed in an inert organic solvent, e.g. dimethylformamide, and at room temperature or at slightly elevated temperature.

The preparation of starting compounds of formula II is either known from the literature, is described in British Patent Specification No. 1,490,852, or can be performed by analogous methods.

In a second step the compounds of formula IV are converted into compounds of the general formulae Va or Vb: in which formulae , Q^, R^, Y, and the dotted line between C-24 and C-25 have the meanings as defined above.

The conversion is performed by reacting a compound of foi'mula IV with a polyhalomethane, such as tetrachloromcthane or tetrabromomothano, or a N- haloamido, If) such as N-chlorosuccinimido, in the presence of triphenylphosphine, a trialkylphosphine, a triaryl phosphite or hexamethylphosphoric triamide, or with an immonium salt of tho general formula VI; i· (0ΙΙ3)2ίϊ=0Η-0-Η4 Y“ VI in which formula R^ represents an unsubstituted or substituted phenyl radical, a phenyloxycarbonyl, alkyl, benzoyl or acetyl radical, and Y- is a chlorine, bromine or iodine ion. The reaction is performed in an inert organic solvent, e.g. ether, tetraliydrofuran, dimethylformamide, acetonitrile, and at or below room temperature.

The compounds of formulae Va and Vb can also be prepared by reacting a compound of formula IV with a phosphorus halide, e.g. phosphorus pentabromide or phosphorus trichloride, with thionyl chloride, or with a (halo— me thylene)-dime thyliminium halide. 1° The conversion of compounds of the general formula IV into compounds of tho formulae Va and Vb normally produces inversion of. configuration at the carbon atom where the substitution takes place (C-l6). Thus, a Compound of formula IV with an α-oriented hydroxyl group at C-l6 .15 is converted into a compound of formula Va, and a compound of formula IV, in which the hydroxyl group at C-16 has β-orientation, is converted into a compound of formula Vb. However, the compounds of formula Va can be transformed into the more stable compounds of formula Vb by reaction with an inorganic or Organic halide, e.g. lithium bromide, tetrabutylammonium bromide, sodium bromide, potassium iodide or sodium - 10 *3 3 4 7 8 iodide*, in an appropriate organic solvent, preferably dimethylformamide, acetonitrile or acetone, at room temperature or at slightly elevated temperature, When a compound of formula XV in which the hydroxyl group at C-16 is α-oriented is reacted in dimethylformamide with an excess of a compound of formula VI, e.g. phenyl Ν,Ν-dimethylformimidato bromide, tho originally formod compound of formula Va is converted into a compound of formula Vb during the reaction.

Tho intermediates of formula Va and Vb are new and. interesting compounds. invention.

Xn a next step the compounds of formula Vb are reac ted with compounds of the general formula VII to form, with lj inversion of configuration at C-l6, compounds of the general formula VIII: VII COOR Λ-R VIII 8 in which formulae , Q^, Iij , It^, and the dotted line bcLweeu C-.'ίύ and C-25 have tiie meaning as defined above, and Λ stands for oxygen or sulphur. If Λ in formulae VII and VIII represents oxygen, the reacting compounds of foi'mula VII are preferably used as solvents, and the reaction is performed in the presence of a silver or mercury salt, e.g. silver carbonate, silver trifluoroacetate or mercuric acetate, or a base, e.g. potassium carbonate, sodium bicarbonate or sodium alcoholate, and at room temperature or at slightly elevated temperature. If A in formulae VII and VIII stands for sulphur, the reaction is performed in an inert organic solvent, preferably ethanol or dimethylformamide, in the presence of a base, e.g. potassium hydroxide or sodium hydride, and at or below room tomporature or at slightly elevated temperature.

In a final step the compounds of formula VIII can be converted into the compounds of formula I by hydrolysis, e.g. in aqueous methanol or ethanol and in the presence of a base, such as sodium or potassium hydroxide or cai-bonate. ι Compounds of formula VIII in which and Qg If stand for the group or 0 and R^ represents an easily hydrolyzable ester radical are without further conversion compounds of the invention. 34 7 8 The compounds of formula VI I.l in which Q and Q„ each JL Tho compounds of formula I in which Q.. and/or n9 stand for oxygen can also be prepared from the corresponding compounds of formula I in which Ω and/or Q,, stand for the group by oxidation methods known to a man skilled in the art.

Compounds of the general formulae I or VIII, in which A stands for a sulphinyl radical are prepared by reacting the corresponding compounds of formulae I or VIII, in which A stands for sulphur, with an oxidizing agent, e.g. hydrogen peroxide, sodium inetaperiodate or chromic acid. The reaction is performed in an inert solvent, e.g. water, acetic acid, ethanol or acetone, at or below room temperature or at slightly elevated temperature.

The easily hydro lysable esters of the compounds of formula .1 can bo prepared in known manner by methods described in tho literature.

Compounds of the invention in which there are single 5 bonds between C-24 and C-25 can also be prepared from tho corresponding unsaturated analogues by reduction, e.g. a catalytic hydrogenation using for instance palladium on carbon as a catalyst.

Intermediates of formula VIII can also be prepared according to one or more -of the following methodst (a). The compounds of the general formula VIII, in which Λ stands for sulphur and R^ is aryl or aromatic heterocyclyl, can be prepared by reacting a compound of the general formula IV, in which the hydroxyl group at C-16 is α-orienfcod, with a phosphine, e.g, tributylphosphine or triphenylphosphine, and a compound of the general formula R^SSR^.

The reaction is performed in an inert organic θ solvent, preferably dimethylformamide or pyridine, and at or below room temperature. - 14 43478 (b) Xn another method a compound of formula XV, in which the hydroxyl group at C-16 is ) Ils α-oriented and is different from the group jjq..-1' i is reacted with a reactive derivative of an, alkyl5 sulphonic or arylsulphonic acid, e.g. an acid chloride or acid anhydride, to form a compound of the general formula IX: in which Q2, R^ and the dotted line between C-2lt and C-25 are as dofinod above for compounds of formula IV, ι If. stands for oxygen or the group Rp being an alkanoyl, aralkanoyl or aroyl radical, and R^ represents an alkylsulphonyl or arylsulphonyl radical, in particular a methanesulphonyl or a p-toluenesul15 phonyl group.

In a next step a compound of formula XX is reacted with a compound of the general formula VI i to form a compound of the general formula VIII. If A in formulae VII and V11T stands Cor oxygen, the compounds of formula VII can be used as solvents, and tlie reaction can be performed at room temperature or at slightly elevated temperature, in some cases in the presence of an organic base, e.g. triethylamine. tf A in formulae VII and VIII represents sulphur, the reaction can be performed in the same way as described above for tho conversion of compounds of formula Vb into compounds of formula VIII, in which A stands for sulphur Ιθ (c) In a further method compounds of the general formula VIII, in which A stands for oxygen, sulphur or a sulphinyl radical, and represents a hydroxy-substituted alkyl radical, can be converted into corresponding compounds in which R^ stands for a halogen -substituted alkyl radical by methods describod above for tho conversion of compounds of formula IV into compounds of formulae Va or Vb.

In a’ following step, the halogen -substituted alkyl derivatives of formula VIII can be reacted with an aliphatic or aromatic alcohol, preferably in the presence of a silver salt or a base, with an aliphatic 43473 or aromatic mercaptan, preferably in the presence of a base, with ammonia or an aliphatic or aromatic amino, or with salts of lower alkanoic acids or benzoic acid, with silver or sodium fluoride, alkali metal azides, nitrites, cyanides or thiocyanates, or with salts of lower thioalkanoic acids or thiobenzoic acid, to form compounds of formula Vi CI in wliich stands for an alkyl radical substituted by e.g. a fluorine atom, an alkyloxy, aralkyloxy, aryloxy, alkylthio, aralkylthio, arylthio, amino, alkylamino, dialkylamino, azido, nitro, cyano, thiocyanato, alkanoyloxy, aralkanoyloxy, aroyloxy, alkanoylthio or aroylthio rail i cal. (d) The C-24,25 unsaturated compounds may in some cases advantageously be hydrogenated to the corresponding saturated intermediates of formula VIII.

The compounds of formula VTII can be converted into the compounds of the invention as already described above.

It is a further object of the present invention to provide pharmaceutical compositions which are useful in the treatment of infectious diseases in the human and veterinary practice.

With this object in view, the compositions of the invention contain as an active component at least one - 17 43478 member selected from the group consisting of compounds of the formula I, salts thoreof with non-toxic, pharmaceutically acceptable bases, and easily hydrolyzable esters together with solid or liquid pharmaceutical carriers and/or diluents.

In the said compositions, the proportion of therapeutically active material to carrier substance can vary from 1% to 95% by weight. The compositions can be worked up to various pharmaceutical forms of presen10 tation, such as granulate, .tablets, pills, dragoes, supjiositorios, capsules, sustained-release tablets, suspensions, injection medicine, or so far as mixtures are concerned, they may be filled in bottles or tubes or similar containers. Pharmaceutical organic or in15 organic, solid or liquid carriers and/or diluents suitable for oral, enteral, parenteral or topical administration can be used to make up compositions containing the present compounds. Water, gelatine, lactose, starch, magnesium stearate, talc, vegetable and animal oils and fats, benzyl alcohol, gum, polyalkylene glycol, petroleum jelly, cocoa butter, lanolin or other known carriers for medicaments are all suitable, while stabilizing agents, wetting and emulsifying agents, salts for varying the osmotic pressure or buffers for securing an adequate pH-value of the composition can be used as auxiliary agents.

Furthermore, the composition may contain other pharmaceutically active components which can appropriately be administered together with the compounds of the invention in the treatment of infectious diseases, such as other suitable antibiotics, in particular such antibiotics, which may enhance the activity and/or prevent development of resistance. Such antibiotic include * penicillins, cephalosporins,' tetracycline, rifamycins, erythromycin, lincomycin and clindamycin.

Other compounds which advantageously may he combined with tho compounds of the invention, especially in topical preparations,include e.g. corticosteroids, such as hydrocortisone, triamcinolone or fluocinolone.

For granulates, tablets, capsules or dragees the pharmaceutical composition of the Invention appropriately contains from 25 per cent to 98 per cent of the active substance of the invention, and in oral suspensions the corresponding amount is appropriately from 2 to 25 per cent.

For parenteral use the compounds are preferably given by intravenous infusion of an aqueous solution - 10 4 3 4 7 8 containing from 0.1 to 2 per cent of the active ingredient, or the compound might be given by injection of the compounds in pharmaceutical compositions with from 1 to 20 per cent active ingredient.

When the compounds are administered in the form of salts with pharmaceutically acceptable non-toxic bases, tho preferred salts are for instance the easily water-soluble sodium salts or tho diethanolamine salts, but other pharmaceutically acceptable ahd non-toxic salts may be used, for instance salts which arc slightly soluble in water, in order to obtain a particular and appropriate rate of absorption.

As indicated above, the compounds of formula I and their salts may he worked up to pharmaceutical forms of presentation including suspensions, ointments and croains. A pharmaceutical preparation for oral treatment may also be in the form of a suspension of a compound of formula I as such or in the form of a sparingly soluble salt with a pharmaceutically acceptable base, the preparation containing from 20 to 100 mg per ml of vehicle.

A pharmaceutical preparation for topical treatment may be in the form of an ointment or cream containing a compound of formula X in an amount of from 0.5 to 10 g per 100 g of preparation.

C 4 Ί S Another object of the invention resides in the selection of a dose of tho compounds of the invention which dose can be administered so that the desired activity is achieved without simultaneous secondary effects 5 In tho human systemic therapy, the compounds and their salts arc conveniently administered (to adults) in dosage units containing not less than 50 mg and up to 1000 mg, preferably from 200 to 75° mg, calculated as the compound of formula I.

By the term dosage unit is meant a unitary, i.e. a single dose which is capable of being administered to a patient, and which may be readily handled and packed, remaining as a physically stable unit dose comprising either the active material as such or a mixture of it with solid or liquid pharmaceutical diluents or carriers .

In the form of a dosage unit, the compound may be administered one or more times a day at appropriate intervals, always depending, howovor, on the condition of the patient, and in accordance with the prescription made by the medical practitioner.

Thus in systemic treatment a daily dose will preferably be an amount of from 0.5 to j g of a compound of formula I. 4 7 8 The torm dosage unit in connection with topical use means a unitary, i.e. a single dose capable of being administered topically to a patient in an application per sq. centimetre of the infected, area of from 0.1 mg_to 10 mg and preferably from 0.2 mg to 1 mg of the compound in question.

If the composition is to be injected, a sealed ampoule, a vial or a similar container may be provided containing a parenterally acceptable aqueous or oily injectable solution or dispersion of the active material as the dosage unit.

The parenteral preparations arc in particular useful in the treatment of conditions in which a quick response to the treatment is desirable. In the continuous therapy of patients suffering from infectious diseases, the tablets or capsules may be the appropriate form of pharmaceutical preparation owing to the prolonged effect ohtained when tho drug is given orally, in particular in the form of sustained-release tablets.

In the treatment of infectious diseases, such tablets may advantageously contain other active components, as mentioned hereinbefore.

Still another object of the invention is to provide a method of treating patients suffering from infectious diseases, the method comprising administering to adult — 22 patients from 0.25 ε to 4 g por day, preforably from 0.5 to 3 g per day, of a compound of the formula X or an equivalent amount of a salt as defined before of a compound of the formula I. Preferably, the compound is given in the form of the dosage units aforesaid.

In the following are given some examples of the preparation of intermediates which are illustrative but not limiting for the invention. .?'! 43478 Preparation .1 3-0-Ace tyl-l6-deacet.oxy-l6a-bromofus idle acid phenacyl · ester A. 3-0-Acetyl-l6-cpitJeacetylfusidic acid phenacyl ester Tho sodium salt of 3-0-aeetyl-J 6-opidoacetyl fus i die aei B. 3-0-.Acetyl-16-dcacetoxy-16q-bromofusidlc acid ; phenacyl ester A suspension of dimethylformamide (l.l ml; 40 mmol) and phenyl chloroformate (5.04 ml; 40 mmol) in 80 ml of petroleum ether (Bp. < 50°c) was stirred vigorously at room temperature. During ono hour carboplienoxy N,Ndimothylformimidato chloride was formed as colourless crystals. This initial product lost carbon dioxide on further stirring for 16 hours to give crystalline phenyl N,N-dimethylformimidate chloride. This in turn - 24 •13 4 7 8 was conver t ι cl into .V, X'-dimothyl f'oimami.de diphenyl acetal by adding a solution of phono) (3.76 g; ;IO nmol) and triethylamine (5.5b mi; 40 mmol ) in oilier (10 till) to the stirred reaction mixture. After stirring for an additional, hour, the triethylamnionium chloride, foinicd ns a by-product, was filtered off and washed with 50 ml. of petroleum ether. When acetyl bromide (,?.<,> ml; 27 mmoj ) was added with stirring to the combined filtrate and washing, phenyl N,N-dimethylforinimidate bromide was formed as colourless crystals, which were collected and washed with petroleum ether (20 ml) to remove traces of phenyl acetate.

The immonium bromide thus obtained (6 g; 26 mmol) was very hygroscopic, and was immediately added to a solution of 3-O-aectyl-l6-epideacetylfusidic acid phenacyl ester (6.2 g; 9.8 mmol) in dimethylformamide (4o ml). After standing fox· 48 hours at room temperature, this solution was diluted with ether (100 ml), washed with 0.1 N sodium hydroxide (100 ml) and water (3 x 50 ml), dried, and evaporated in vacuo. Addition of methanol (50 ml) caused the residue to crystallize. The crystals were filtered off, washed with methanol, and dried te afford 5·- β 3-0-acetyl-J.6-deacotoxy-l6«-bromofusidic acid phenacyl ester, melting point l)l-l42°C.

Proparn bioils 2-8 3-O-Ac.csfcyi-l6-deacc 1 oxy-l6tt-broinorusidic acid ostfrrs Λ. Dy following Uns procedure of Preparation 1 A but substituting tho esterifying agents shown in table I for the phenacyl bromide, the 3-O-acetyl-l6epideacetylfusidic acid esters indicated in table I were obtained.

B. By substituting the 3-0-acetyl~l6-epidoacetylfusidic acid esters indicated in table X for the 3-0 acetyl-l6-epideacetylfusidic acid phenacyl ester in ✓ the procoduro of Preparation 1 B, the 3-0-acotyl-l6deacetoxy-l6a-bromofusidic acid esters shown in tahl I were obtained.

Tabic I; Pre paration Es t. r i f y i. ng age 111 Resulting compounds}!1K2 Mp (f) 2 Λ oh l.oromo thyl acetate OH ciuococn,. f- J amorphous 2 B Br CH?OCOCH3 102-105 3 A chioromethyl pivalate OH CH?0C0C(CH3) amorphous 3 B Br CH,,OCOC(CH3) amorphous 4 A chioromethyl. benzoate OH Cll^OCOCJf2 6 5 amorphous 4 B Br C!K0C0C,H_ 2 6 a 93-94 5 A chloroacetonitrile OH CH CK amorphous 5 n Br CIlpCN I22-J.23 6 A benzyl bromide OHCU2C6n5 108-100 6 B BrCH2C6n5 128-129 7 a p-methyl benzyl bromide OH ch2c6h4ch3 amorphous 7 B Br ch2c6h4ch3 amorphous 8 A chi 0rome thyl methyl e ther OH CH,OCH3 amorphous 8 n Br ch2ocii3 amorphous Preparation Q 3-O-Act> tyl-1 fa-deacotoxy-Kiit-bromofusld i.e acid p-nitrobenzyl ester The sodium salt of 3-O~acetyl-l6-epideacetylfusidic acid (21.5 S5 40 mmol) and p-nitrobenzyl bromide (9.5 gi 44 mmol) were dissolved in dimethyl formamide (200 ml). * — 27 — 3 478 'Mils solution'vas left at room temperature for 16 hours, during which period 3-O-acetyl-l6-epidcacetylfusidie acid p-nitrobenzyl ester was formed. Phenyl N,Ndimethylformiinidate bromide (36 g; see Preparation 1 B for tho preparation of this reagent) was then added, and the resulting red-brown solution was kept at room temperature for 48 hours. Methanol (700 ml) and water (280 ml) were added with stirring to precipitate a crystalline product. The crystals were filtered off, washed with methanol:water 3sl> and dried to afford 26.1 g of 3-0-acetyl-l6-deacetoxy-ljja-bromofusidio acid p-nitrobenzyl ester, melting point; 151-157°C. Recrystallization' from methanol;water gave the analytically pure compound, melting point: 157-159°c· Ij Preparations 10-12 3-0-Acotyl-l6-dcacetoxy-l6(X-bromofusldic acid esters By following the procedure of Preparation 9 but substituting the esterifying agents listed in table II for p-nitrobenzyl bromide, the 3-0_acetyl-l6-deacetoxy20 Ιόα-bromofusidic acid esters shown in table II were obtained.

Table 11; Y - 28 43478 I’ropa ra I i on I s ! eri !'y leg agen 1 Hi-Mil i « η,·; compounds K Mp (C) 10 p-benzylphenacyl bromide CH,,COO, Jl,.011,,0,Hc 2 6 0 2 6 5 127-J29 * 1 p-me thoxy pheimcy 1. lu-oiii i do 011,,000, II. OOH,. 2 () '1 J 114-116 12 bromoacetono 1 CII^GOCH^ 80-81 Preparation 13 3-0—Acetyl -16—deaeotoxy-l6o;—broinofus j cl lc acid benzyl ester A. 3-0-Acetyl-l6-dcacotylfusidic acid benzyl ester.

To tiie sodium salt of l6-deace±ylfusidic acid (84,7 g; 0.17 mol) in dimethylformamide (200 ml) was · added benzyl bromide (25 ml; 0.21 inol). After stirring for 5 hours at room temperature, tho resulting io solution was cooled to 0°0, and pyridine (200 ml; 2.5 mol) and acetic anhydride (170 mi; 1.8 mol) were added. After standing for .16 hours at room temperature, the mixture was again cooled to 0°0, and 50 ml of water was added with stirring at such a rate, that the tom15 porature remained below 15°C (about 1 hour was required for this addition). Then methanol (800 ml) and water (400 ml) were added to complete the precipitation of tho desired product, which, after stirring for 1 hour at 10°C, was filtered off, washed with ice-cold me thanol (') x 20 mJ ), and dried to yield 68 g of 3-0-acet-yl-16-deacetylfusidic acid benzyl ester as colourless crystals, melting point 154-158°C.

B. 3-0-Acotyl—16-doncc boxy-l6(X-broinofu»idic acid benzyl ester The above benzyl ester (68 g; 112 mmol), sodium bromide (46.2 g; 448 mmol) pyridine (22 mlj 276 mmol) and dimethylformamide (400 ml) was stirred for 30 minutes at room temperature and then cooled to 0°C. Plionyl chloroformato (56.5 ml; 448 mmol) was added over a period of 45 minutes, and thp resulting mixture was stirred at room temperature for 18 hours. After, this period, the reaction product was precipitated by addition of methanol (4θθ ml) and water (300 ml) as colourless crystals, which were filtered off, washed with methanol;water (2 χ 6θ ml of a 2:1 mixture) and petroleum ether (3 x 30 ml), and dried to yield 62.9 g of 3-0-acotyl-l6-deacetoxy-l6a-bromofusidic acid benzyl ester, melting point 124-126°C.

Preparations 14-15 3-0-Acetyl-l6-deacetoxy-l6g-hroniofusidic acid esters A. Following the procedure of Preparation 13 A but substituting phenacyl bromide or p-nitrobonzyl bromide for the benzyl bromide, tho 3-0-aoetyl-l6-deacetylfusidic acid esters indicated in table III were obtained B. By substitut. ing tho 3-0-acotyl-l(>~doacelylfusidie acid esters siiiiwn ia tabic Ilf for tin· 3-O-acotyl—J6— draco tyl fusidic ao:id benzyl ester in tho procedure of Preparation J 3 B, tho 3-0-acetyl-l6~dcacetoxy5 16α-bromofusidic acid esters indicated in table 111 wore obtained.

Table III : K .·* ii Resulting compounds PreparationR1R2 Mp (°C) 1.4 A β-ΟΗ CH„COCrHe 2 6 5 149-151 1.4 B a-Br cb2coc6h5 141-142 15 A β-ΟΗCII2C6H4N°2 141-143 15 B a-BrCH2C6H4N°2 157-159 Preparation 16 3-0-Acety1 - 1 (i-doacetoxy- ]6rt-bromofus Idle acid piva 1 ey ί oxymotliyl ester A, 'i-O-Aco tyl —16—doneoto\y-1 66-broiuofiis id ic acid Ρ1valoy loxymothyl os ter 3-0-Acotyl.-16-ep.idoacotylfusidic acid pivaloyloxyme l.hyl ester (17.4 g· 28 mmol) was dissolved in dry other (200 ml), and triphenylphosphine (l6 g; 60 mmol) and tctrabromomothane (20 g; 60 mmol) was added.

Aftei' stirring for 16 hours at room temperature, tho reaction mixture was filtered to remove triphonylphospliine oxide, which was formed as a by-product.

The filtrate -was ,evaporated in vacuo, and the residue was purified i>y dry column chromatography on silica gol (cyclohexane:ethyl, acetate 8:2) to give 10.6 g of 3-0-acetyl-l6-deacetoxy-l6p—bromofusidic acid pivaloyloxymethyl ester as colourless ci'ystals, obtained from ether-petroleum ether,_molting point 120-122°C. Itocrystallization from ether-petroleum other afforded the analytically pure compound, melting point 120-122°C.

B. 3-0-Acetyl-l6-deacetoxy-l6Q:-bromofUsidlc acid pivaloyloxymethyl ester The 3-0-acotyl-l6-deacetoxy-l6p-bromofusidic acid pivaloyloxymethyl ester described above· (5 g) was epimerized to the l6a-compound by reacting with tetrabutyXaimnonium bromide (5 g) in acetonitrile (60 ml) for throe days at room temperature. Tho reaction mixture was evaporated in vacuo, and ether was added to tlie residue causing tetrabutylammonium bromide to y cryslallize, The crystals were filtered off, rttid tin· filtrate was washed with water (2 x ill till), dried, arid evaporated i n vacuo to yield h.87 g of 3-0-acetyll6-doacotoxy-l6a-bromofusidic acid pivaloyloxymethyl S ester as a colourless gum.

Propa ra t. i on 17 3-0-Ace I y I -16-deacct.oxy-J 6a-broinofusi.dlc acid acetoxymethyl ester A. 3-0-Acetyl-l6-deacetoxy-l6P-broniofusidic acid * acetoxymetliyl ester By following tho procedure described in Preparation 16 A but substituting 3-O-acetyl~l6-cpi.deacetylfusidic acid acetoxymethyl ester for the 3-0acetyl-16-epideacetylfusidic acid pivaloyloxymethyl ester, 3-0-acotyJ.-l6-doacotoxy-16(i-broinofusidic acid acetoxymetliyl ester was prepared, tiioLLing point 119—120 0.

B. 3-Q-Acotyl-1.6-doiicetoxy-l6(X-broinofusidlc acid acetoxymethyl ester By following the procedure described in Preparation 20 1.6 B but substituting 3-0-acetyl_l6-deacetoxy-l6p-biOmofusidic acid acetoxymethyl ester for the 3-O-acetyl-l6deacetoxy-J.6p-bromofusidic acid pivaloyloxymethyl ester, 3-0-acetyl-l6-deacetoxy-l6a-bromofusidic aeid acetoxymethyl ester, melting point 102-105°C, was prepared.

Preparation 18 3-0-Ano fcyi-l6-dcacetoxy—l6a-bromo-24,25-dihydrofusldlc acid phenacyl ester A. 16-Doncotyl—2/l ,25-dlbydrofusidic acid phenacyl ester To a solution of l6-deacetyl-24,25-dihydrofusidie acid sodium salt (4.99 g; 10 mmol) in dimpthylformamido (25 ml) was' added phenacyl bromide (i.99 g; lumo1), and tho mixture was stirred at room tom10 perature for 4 hours. After dilution with ether (100 ml), the mixture was washed wJ,tli water (4 x 25 mi). The organic phase was separated, dried, and coneon— tratod to about 20 nil whereby a crystalline product precipitated. After being kept in the refrigerator for 2 hours, the crystals were filtered off, washed with ether, and dried to afford 4.52 g of thp desired compound, melting point 92-94°C. (dec.). 1). 3-0-Acetyl-l 6-do.-ir.ety1 -24,25-dihydro fusid i c acid phenacyl ester 2o To a stirred solution of 16-deacetyl-24,25-dihydrofusidic acid phenacyl ester (2.38 g; 4 mmol) in pyridine (8 ml) was added acetic anhydride (4 ml), and . the mixture was left at room temperature for 16 hours. After dilution of the stirrod reaction mixture with diisopropyl ether - 34 43478 (60 ml), crystallization of a colourless product occurred. ΊΙιο inyhtnls were collected, washed with d i I sojirepy 1 ether and dried to yield 1.02 g of the desired compound, melting point 133-135°θ· C , 3—0—Act' ty I — I Phenyl chloroformate (l.26 ml; 10 mmol) was added dropwise at 0°C to a stirred solution of 3-θacetyl-l6-doacetyl-24,25-dihydrofusi.die acid phenacyl ester (l.59 gj 2.5 mmol), sodium bromide (l.03 g; 1.0 mmol), and pyridine (0.52 ml; 6,5 mmol) In dimethylformami.de (15 ml). After the addition was finished («^15 min.), the reaction mixture was stirred for 2 hours at 0°C, followed by 16 hours at room temperature. Dropwise addition of methanolswater ltl (15 ml) to the stirred mixture precipitated a crystalline product which was filtered off, washed with methanol, and dried to give 1.22 g of the desired compound, melting point 127-129°C. Recrystallization from methylene chloride-methanol gave tho analytical sample, melting point 13O-132°C, Preparations J 9-20 3-0-Ace tyl-1 6-doace toxy- I fia-brotno-24,2 5-d i. hydro fusldie acid esters Hy substituting benzyl bromide or p-nitrobenzyl bromide for tlie phenacyl bi’oinide in tho procedure of Preparation 18 Λ, tho l6-doacetyl-24,25-dihydrofusidic acid esters indicated in tabl.e IV below were obtained.

B, Following the procedure of Preparation 18 B, but substituting the l6-deacetyl-24,25-dihydrofusidic acid esters shown in table IV for the l6-deacoty1.-24,25· dihydrofusidic acid phenacyl ester, the 3-0-a-ootyl ester derivatives indicated in table IV were obtained.

C. Following the procedure of Preparation 18 C, but substituting the 3-0-acetyl-l6-deacetyl-24,25-dihydrofusidic acid esters shown in table IV for the 3-0aeetyl-l6-doacotyl-24,25-dihydrofusidic acid phenacyl ester, tho 3-0-acotyl-l6-deacetoxy-I6a-bromo-24,25di hydrofusidic acid esters indicated in table XV wore Ij obtained.

Table IV : ·- - 36 43478 Proper 1 L j oil Resulting compounds «1 «0 «3 MP (°C) 19 Λ II fi-0IIC112C6H5 amorphous 19 B CII co [j-OH oh2c6h5 162-103 19 C CH^CO CC-Br ch2c6b5 104-103 20 A II β-ΟΙΙ ch2c6h4no2 amorphous 20 II CII3C0 β-OIJ. ch2c6h;(n°2 amorphous 20 C CH^CO OC-Bi’ CH2C6H/tNO2 147-1.49 Preparation -21 3-0-Forniyl-l^-doaeetoxy-lGg-bromofusldie ac i d benzoyl oxymethyl ester A. 16-Epidoacety1fusidic ac i d benzoyl oxymot-hy 1 osier I.6-Epidcacotylfusidic acid (35.5 gj 75 mmol) was dissolved in methanol (150 ml) and converted into its sodium salt by titration with 5 N sodium hydroxide using phenol phthaJoin as indicator. After evaporation to dryness in vacuo, the resulting amorphous sodium salt was dissolved in dimebhylformainido (150 ml), chloromethyl benzoate (l4.O8 g; 82.5 mmol) was added, and the mixture was stirred at room temperature for 16 hours. Water (200 ml) was added, and the mixture was extracted with ether (1)00 ml). The organic phase was separated,' wasiied with water (4 x 100 ml), driod, and evaporated In vacno to yield 44.6 g of 16-epideacetylfusidic acid benzoyioxymethyl ester as an amorphous product.

B. ^-O-Ferinyl-l6-deaeetoxy-l6o:-broinofusidic acid benzoyloxymethyl ester The 16-epideacetylfusidic acid benzoyloxymethyl ester prepared above was dissolved in dimethylformamide (300 ml), plienyl N, N-d ime tliyl formin'· Ida Le bromide (67 g; ea. 290 mmol ) was added with sl,.i.rripg, and the rodbrown solution was kept at 5°C for 6-7 days. To the mixture was added methanol (150 ml), and, with vigorous stirring, water (l50 ml) from a separating funnel to precipitate a crystalline pi'oduct. The crystals were filtered off, washed with methanol:wator lsl, and dried to afford 27.1 g of 3“0-formyl-l6-deacetoxyl6a-bromofuSidic acid benzoyloxymethyl ester, melting point 131-135°θ· Two recrystallizations from ethermethanol raised the melting point to l40-l42°C, Preparations 22-24 3-0-Formy]-l6-deacetoxy-l6a-hroinofusidlc acid esters , A. By substituting benzyl bromide, chloromethyl * _ - 38 43478 pivalate or ehl oromoUiyl acetate for the chi. or omethyl benzoate in the procedure of Preparaiion 21 A the 16epideacetylfusidie acid esters indicated in table V below were obtained.

B. Following tbe procedure of Preparation 21 B, but substituting the 16-epidoacetylfusidlc acid esters shown in table V for the l6-epidoacetylfusidie acid benzoyloxymethyl ester, the 3-0-formyl-l6deacetoxy-l6a-bromofusidic acid esters indicated in table V were obtained.

Resulting compounds PreparationRiR2R3 Mp (°C) 22 A H OH ch2c6h5 95-98 22 B HCO Br ch2c6h5 125-127 23 A H OH CH?OCOC(CH3)3 amorphous 23 B HCO Br Cil2OCOC(CH3)3 amorphous 24 A H OH CH2OCOCH3 amorphous 24 B HCO Br CJI,,OCOCH 123-125 Preparation 25 3-0-Foniiy.l -iG-denCP toxy-1 btt-bromof itsldi c acid benzyl ester A. .t6-T)eacetyl fus idle. acid benzyl ester To fi solution of 16-deacetylfusidic acid sodium salt (4.97 g; 10 mmol) in dimethylformamide (25 ml) was added benzyl bromide (1.5 ml; 12.5 mmol), and the mixture was stirred at room temperature for 4 hours. After addition of water (100 ml), the mixture was extracted with ether (2 x 50 ml), and the combined organic extracts were washed with water (4 x 20 ml), dried, and evaporated in vacuo. The residue thus obtained was dissolved in ether (50 ml), and on addition of petroleum ether (5θ ml) with stirring a crystalline product precipitated. The crystals were filtered off, washed with ethers petroleum ether 1:2, and dried to give 4.92 g of the desired compound, melting point 117-119°C.

B. 3-0-Formyl-l6-deacntylfusidic acid benzyl ester 2θ Acetic formic anhydride (4 ml) was added dropwise at 0°C to a stirred solution of 16-deacetylfusidic acid benzyl ester (4.52 g; 8 inmol) in pyridine (8 ml), and tho mixture was kept at the low temperature for 15 minutes. On. dilution of the stirred reaction mixture • - - 40 43478 with <1 iisopropyl ether (40 ml), a crystalline product precipitated. After being kept in the refrigerator for 2 hours, the crystals were collected, washed with diisopropyl. ether, and dried to yield 4.04 g of the desired compound, melting point l4,3-l45°C. Recrystallization from ethor-diisopropyl ether afforded the analytical sample', melting point l4.5-l47°C· C, 3-0-l''orniyJ.-3.6-deacetoxy-l6g.-broinofusidic acid benzyl ester, Dy following tlie procedure of preparation 13 B but substituting the above 3-0-formyl-.l6-deacetylfusidie acid benzyl ester for the 3-O-acetyl-l6deacetylfusidic acid benzyl ester, 3-0-formyl-l6dcacetoxy-l6a-broinofusidie acid benzyl ester was prepared as colourless crystals; melting point 125-I27°O.

Preparation 26 3-0-Forniy I -1 h-deacotoxy-l6a-broi)iofusidle acid pi valoy Ιο xymethyl ester A, ]6-Beacetylfusidie acid pivaloyloxymethyl. ester To a solution of the amorphous silver salt of 16deacetylfusidie acid (5*8 g; 10 mmol) in dimethylformamide (50 ml) was added chlorometliyl pivalate (l.48 ml; 10 mmol), and the mixture was stirred at - 4i 43478 room temperature for 4K hours. Filter aid was used to remove insoluble material, which was washed with ether (2 x 25 ml). The combined filtrate and washings were diluted with ether (100 ml), the resulting mixture was washed with water (4 x 50 ml), and the organic phase dried and evaporated in vacuo to afford the crude ester as a yellowish foam. Purification of the residue by dry column chromatography bn silica gel (developing .solvent: Cyclohexane:ethyl acetate 3:7) yielded the desired ester as an amorphous product ·» which failed to crystal i I ize, Tho NMR spectrum ('COCl') shows signals at = 0.90 (d,3H), 0.93 (s, 3H), 0.98 (s, 311), 1.22 (s, 911; CfCH^) ), 1.38(s, 3»), 1.62 and 1.68 (2 bs, 6ll), 2.99 (m, IH; CH-13), 3.77 (m, IH; CH-3), 4.33 (m, IH; CH-ll), .00 (m, IH; CIT-Ιό), 5.12 (ni, IH; CH-24), and 5.15 and 5.42 (2 d, J=7, 2H; ‘ OCH,,0) ppm. Tetramethylsilane was used as internal reference.

B. 3-0-Fonnyl-l6-deacetylfusidic acid pivaloyloxy20 methyl ester Following the procedure of Preparation 25 B, but substituting 16-deacetylfusidic acid pivaloyloxymethyl ester for the l6-deacetylfusidic acid benzyl ester, the 3-0-formyi-l6-doae'etylfusidic acid pivaloyloxymethyl ester was obtained. - 42 43478 0. 3-(1-1-11 nny I - I fi-denrn tnxy-l 6h-li I’QllKi fns id i c ar. i-tl p i va 1 ny 1 oxyini-1 hy 1 ns tor lly subs I i tuting 3-0-formyi -16-doncetylfusidic acid pivaloyloxymethyI ester for tJio 3-0-acotyl5 I 6-doaeofy 1.1'ijm i d i o aeid benzyl ester in the procedure oi- i*i-oparati.on .13 H, 3-0-fo riuy 1.-J.6-doacetoxy-.1.6abromofusidle acid pivaloyloxymethyl ester was obtained as a colourless foam.

The NMli spectrum (CIJCl^) shows signals at J =: 0.78 10 (s, 3H), 0.87 (d, J=7,3H), 1.00 (s, 3H)| 1.23 (s, on, C(cn3)3), 1.4? (s, 311), 1.61 and 1.68 (2 bs, 6ll), 3.45 (m, IH; CH-13), 4.35 (m, IH; CH-ll), 5.08 (m, III; CII-3), 5.12 (m, III, CH-24), 5.62 (bt, lit, 0Π-16), 5.82 and 5.92 (2 d, J=6, 211; OCHgO), and 8.15 (bs, IH, ΠΟΟ) ppm. Tetramethylsilane was used as internal roforone»·.

Preparation 27 3-0-Pormyl-16-deacetoxy-lfiq-bromo-24,25-diliydrofusidic acid acetoxymethyl ester 20 A. 16-BpidoacotyJ —24, 25-dihydrofasi die acid acotoxymothyl ester To a solution of l6-opidoacetyl-24,25-dihydrofusidic acid potassium salt (20,6 g; 4o mmol) in dimethylformamide (ΐόθ ini) was added chloromethyl - 43 43478 acetate (4.0 mi; .44 mmol), and the mixture was stirred at room temperature for 18 hours. After dilution with ether (500 ml), the mixture was washed with water (2 χ 150ml, 4 x 75 ml), and the organic layer was dried and evaporated in vacuo to give the desired compound as a colourless foam.

B. 3-0—Formyl—l6-deaee fcoxy-l6(t-hromo-24, 25-Qibydro— fusidlc acid acetoxymethyl ester To a stirred solution of the above ester (40 mmol and sodium bromide (20.6 g; 0.2 mol) in dime thy If ormamidc (200 ml) was added dropwise at 0°C phenyl chloro formate (25.2 ml; 0.2 mol). After the addition was finished (about 45 minutes), the mixture was stirred at 0°C for 3-A hours and at room temperature for a further 10-12 hours. Precipitated sodium chloride 'was filtered off and washed with dimethylformamide (2 x 25.ml). To the combined filtrate and washings was added methanoltwater lsl (300 ml) with stirring to precipitate a crystalline product. The crystals wore filtered off, wasiied with methanol;water 1:1, dried, and finally recrystallized from ether-diisopropyl ether to afford 15.35 g of the desired compound melting point 126-127°C. - 44 43478 Proparat ions 28-32 I 6-l1e.'iee toxy- I 6ι·-|ι ioiiio I’usi d i e acid os tors By substi tut i.ng J.6-epideacetylfusidic acid benzyl, phenacyl, pivaloyloxymethyl, acetoxymethyl 5 or benzoyloxymotliyl ester for the 3-O-aeetyl-l6epidoacetyhfusidie acid pivaloyloxymethyl ester in the procedure of Preparation 16, the 16-deacetoxy-J6ocbromofusidic acid estors listed in table VI were obtained.

Table VI ! Resulting compounds Preparation R Mp(°C) 28CH2C6H5 amorphous 29 CHnCOCrHc *2 6 5 amorphous 30 CH,,OCOC(CH )„ ** J J amorphous 31 CHgOCOCILj 105-106 32 ch, ococriie 2 6 5 amorphous - 45 4347 8 { Pi^eparai i on 33 3-Q-Aeoiyl-l1- dehydro-l6-deacetoxy-l6a-brom0fusidic acid, phenacyl ester To a solution of 3-0-aoetyl-l6-deacotoxy-l6a5 bromofusidic aci.d phenacyl ester (6,98 g; 10 mmol) in acetone (7θ ml) was added Jones reagent (3«0 ml), and tho mixture was stirred for 3θminutes at room temperature. After dilution with other (lOO ini) and addition of water (70 ml), the mixture was stirred for a further 10 minutes. The organic layer was separated and the aqueous^ layer reextracted with ether (lOOml). Tho combined organic extracts were washed with water until neutral, dried, and concentrated to about 50 ml, whereby precipitation of a colourless crystalline product occurred, After being kept in the refrigerator for 1 hour, the crystals were filtered.Off, washed with ice-cold ether, and dried to give 5·37 S of 3-0-acetyl-ll-.dehydro-16-deacetoxyl6a-biOiiiofusidie acid phenacyl ester, melting point 120-121°C. On concentration of the mother liquor another 0.95 S °f the desired compound, melting point ll4-ll6°C, was obtained. Recrystallization from methylene chloride - diisopropyl ether afforded the analytical sample, melting point 120-121°C. - 46 43478 Preparation 34 3-()-AcetyJ -1 I - dehydro-16-deacetoxy-l 6 ot-bromofusidic acid benzyl ostor By following tho ijrocedure of Preparation 33 and substituting 3-0-acetyl-l6-deacetoxy-l6a-hroinofusidic acid benzyl estor for 3-0-acetyl-10~deacetoxyl6a-broniofusidlc acid phenacyl estor, 3-0-acetyl-ll. dehydro-l6-deacetoxy-l6«-brofflofusidic acid benzyl ester was prepared as a colourless foam.

Tho NMlt spectrum (CBOl^) shows signals at ^=:1.00 (s, 311), 1.05 (s, 3Π), J .0.2 (d, 3H), 1.27 (s, 3»), 1.62 and 1.68 (2 bs, 611), 2.06 (s, 3Π; CH^CO), 3.30 (m, 111; CH-13), 4.95 (m, IB, CII-3), 5.05 (m, IH, CH-24), 5.22 (s, 211} CH2C6H5), 5.60 (bt, IH} CH-Ιό), and 7.35 (s, 5H5 aroin. CIl) ppm. Tetramethylsilane was used as in to i-ual reference.

Preparal ion 35 3-0-Forinyl-l 1 -dehydro-16-deacetoxy-l6a-bromo-24,25-dihydrofusldic acid acetoxymethyl ester By substituting 3-0-formyl-l6-deacetoxy-l6a-bromo24,25-dihydrofusidic acid acetoxymethyl ester for the 3-0-acetyl-l6-deacetoxy-l6a-broinofusidic acid phenacyl ester in the procedure of Preparation 33» 3-0-formyl-ll- 47 43478 dehydro-16-dcacetoxy-l6 a-bromo-24,25-dihydrofusidic acid act· (.oxymotliyl osier was obtained as a colourless foam.

The NMR spectrum (CDGl^) shows signals at J = 0.87 (d, J=5.5, 611), 1.02 (s, 311), l.o4 s, 3H), 1.25 (s, 3H), 2,12 (a, 311; CI^CO), 3.35 (m, IH; CiI-13), 5.10 (m, 111; Cir-3), 5.68 (bt, III; CH-16), 5.81 and 5.90 (2 d, J=5.5, 211; OCHgOj, and 8,15 (bs, IH; HCO) ppm. Tetramethylsilane was used as internal reference.

Propara 11on 36 - «* .l-Dehvdro-l6-dcacetoxy-l-6a-bromofusidic acid acetoxy- . methyl ester A. fl-PohydEp·,! 6-cpidcacctyl Cnsi die, ani d acetoxymethyl ester To a solution of 3- dehydro-16-epideacetylfusidic ij acid potassium salt (3.06 g; 6 mmol) in dimethylformamide (30 ml) was added eliloromethyl acetate (0,6 ml; 6.6 mmol), and tbe mixture was stirred at room temperature for 18 hours. Tho reaction mixture was diluted with ethe-r (1.00 ml) and washed with water 2θ (4 x 30 ml). The organic phase was separated, dried, and evaporated in vacuo to afford 3.2 g Of the desired compound as a colourless foam. _ 48 43478 Β· 3-Dehydro-l6-deacetoxy-l6n-bromofusidic acid ac ο 1: oxymethyl ester By following tho procedure described in Preparation 27 B but substituting the above 3-dehydroj 16-epideacetylfusidic acid acetoxymethyl ester for the l6-epideacetyl-24,25-dihydrofusidic acid acetoxymothyl ester, 3-dehydro-l6-deacetoxy-l6i*-bromofusidic acid acetoxymethyl ester, molting point l44-l45°C, was obtained.

Preparation 37 —3-0-Ace tyl -l6-deneeti)xy-l 6|3-chl orofusl <1 i.e acid methoxymethyl ester 3-0-Acetyl-l6-epidcacetylfusidic acid methoxymethyl ester (1.4 gf 2.5.mmol), triphenylphosphine (2.6 g; 10 mmol) and N-chlorosuccinimide (l.3 gj 10 mmol) were dissolved In dry other (50 ml). After standing for one hour at 35°C, the triphenyIphospli i lie oxide which precipitated was filtered off, and the filtrato was evaporated in vaeuo. The residue was puril'iod by dry column chronia20 tography on silica gel (cyclohexanesetliyl acetate 7s 3) to afford I.l4 g of 3-0-ncetyl-l6-deacetoxy-l6p-chlorofusidic acid methoxymethyl ester, which was crystallized from petroleum ether, melting point 148-151°C. Recrystallization from cyclohexane afforded the analytically pure compound, melting point l49-151°C. - 49 43478 Preparation 38 .3-0-Ac.etyt-lfi-doacetoxy-l6P-chlorofusidic acid benzyl ester Carbophenoxy Ν,Ν-diniethylforrnimidate chloride was 5 prepared by adding phenyl cliloroformate (θ.3 ml; 2.4 mmol) to N,N-dimothylformamide (15 ml). To tho resulting solution was added 3-P-acotyl-l6-epideacetylfusidic acid benzyl ester (500 mg; 0.82 mmol). After standing for l6-hours at room temperature the reaction mixture was iy . diluted with ether (lOO ml), washed with 2 N sodium hydroxide (25 ml) and water (3 x 25 ml), dried and evaporated in vacuo to give 480 mg of 3-O-acetyl-l6deacetoxy-T6B-chlorofusidic acid benzyl ester, which was crystallized from ether-petroleum ether, melting point Ι63“1ό5°θ· Recrystallization from ethyl acetatepetroleum ethor afforded the analytically pure compound, molting point 165-166°C.

Preparation 39 3-0-Acctyl-l6-deacetoxy-].6g-chiorofusidic acid benzyl ester To a stirred ice-cooled solution of 3-O-acetyl-l6deacetylfusidic acid benzyl ester (1.36 g) in dimethylformamide (lO ml) and pyridine (0.44 ml) was added phenyl chloroformate (1.I3 ml) over a period of 3θ minutes. - 50 4347 After stirring at room temperature for l6 hours, the resulting solution was diluted with ether (lOO ml), washed with 2 N sodium hydroxide (25 ml) and water (3 x 50 ml), dried and evaporated in vacuo. The S residue was dissolved in ether (10 ml) and petroleum ether was added to precipitate the reaction product as colourless crystals, which were filtered off, washed with petroleum other, and dried to yield 3-O-acetyl-l6deacetoxy-l6a-chlorofusidic acid benzyl ester, melting point 115-117°C. Recrystallization from ethyl acetatepetroleum ether raised the melting point to 120-122°C.

Tho invention will be further described in the following Examples which are not to be construed as limiting the invention. 4 34 78 Example 1 l6-Dgacetoxy-l60-isopropylthiofusidic acid Α, 3-0-AeetyI-16-deacetoxy~l6P-isopropylthiofusidic acid p-nitrobenzyl ester 3-0-Acetyl-l6-deacetoxy-l6o:-bromofusidie acid p-nitrobenzyl ester (28.6 g; 40 mmol) was added to a solution of potassium hydroxide (10 g of 8556 purity; 150 mmol) and isopropyl mercaptan (30 ml; 32Ο mmol) in ethanol (1000 ml), and the suspension was stirred for four days. Thereafter, 500 ml of water was added to complete the precipitation of the desired product. The crystals were filtered off, washed with water: ethanol (l:2), and dried to give 21.5 g of crude 3-0-acetyi-l6-deacetoxy-l6f3-isopropylthiofusidic acid p-nitrobenzyl ester, melting point: 157-l6l°C.

B. l6-Deacetoxy-i68-lsopropylthiofusidic acid A suspension of the above p-nitrobenzyl ester in a mixture of ethanol (800 ml) and 2 N aqueous sodium hydroxide (200 ml) was heated to 60°C for three hours.

The resulting dark solution was acidified with 4 N hydrochloric acid (125 ml) and treated for 15 minutes with 5 g of charcoal while still hot. After filtration, 500 ml of water was added, and, after cooling to room temperature, the crystalline product vas filtered off, - 52 424 7 washed with water, and dried to give 14.1 g of 16deacetoxy-l63-isopropylthiofusidic acid, melting point 223-220°C. Recrystallization from 2-butanone gave tho analytically pure compound, melting point: 229-231°C. j Examplos 2-8 l6P-Thloothcrs of l6-deacetoxyfusldic acid A. l6P-Thtoethers of 3-0-acetyl-l6-doacetoxyfusidic acid p-nitrobenzyl estor * By following the procedure described in Example 10 1 A and substituting the mercaptans listed in table VII for isopropyl mercaptan, the Ιόβ-thioethers of 3-0-acetyl-l6-deacetoxyfusidic acid p-nitrobenzyl ester indicated in table VII were prepared.

Table ViT: - 53 43478 Resulting compound Example Mercaptan R Mp (°C) 2 A ethyl mercaptan ch2ch3 167-168 3 A 2-hydroxyethyl mercaptan CHgCHgOH 192-194 4 A 2-aminoethyl mercaptan CEr2CH2Mi2 I88-191 5 A allyl mercaptan CHgCH =CII2 167-170 6 A isebutyl mercaptan CII2CH(CH )2 . 104-112 7 A sec-butyl mercaptan -ch(cii3)ch2ch3 150-157 8 A cyclopentyl mercaptan cyclopentyl 100-109 9 A mercaptoacetic acid methyl· ester ch2cooch3 ·» 125-129 TO A furfuryl mercaptan furfuryl 146-148 B. ' 16R. -Thioethors of 16-deacetoXyfUsidic acid By following tho procedure of Example 1 B and 7 substituting the I6p-thioethers of 3-O~acetyl-l6~ doacotoxyfusidic acid p-nitrobenzyl ester listed in table VII for 3-0-aCetyl-l6-deacetoxy-l6p-lsopropylthiofusidie acid p-nitrobenzyl ester, the l6p-thioethers of 16-deacetoxyfusid.ic acid indicated in table VIII were prepared.

Table VIII; Resulting compounds Example R Mp (°C) 2 B ch2ch3 195-198 3 B CHgCHjjOH 179-182 4 B ch2ch2nh2 241-250 (dec) 5 B ,ch2ch=ch2 196-199 6 B ch2ch(ch3)2 199-202 7 B CH(CH3)CH2CII3 218-222 8 B cyclopentyl 217-223 9 B CHgCOOH 199-202 10 B furfuryl amorphous The NMR spectrum (CDCl^) of’the compound of Example 10 B shows signals at {$ = 0.97 (s, 6h), 1.32 (s, 3H), 1.60 and 1.68 (2 bs, 6H), 3.00 (m, IB; CH-13), 3.73 (m, 1H; CH-3), 3.78 (bs, 2H; SCHg), 4.22 (d, IB; CH-16), 4.30 (m, 111; CH-ll), 5.10 (in, IH; CH-24), 6.1-6.4 (in, 2H; arom. CH) and 7.32 (bs, IH; arom. CH) pptn, Tetramethylsilane was used as internal reference.

Example 11 l6-0eacotoxy-168-isopropylthio-24,25-dihydro-fusidic acid A. 3-0-Acetyi-l6-deacetoxy-l6ft-isopropy.lthio-24,25dihydrofusldic acid p-nitrobenzyl ester By following the procedure of Example 1 A and - 55 43478 substituting 3-0-acetyl-l6-deacetoxy-l6a-bromo- 24,25- dihydrofusidic acid p-nitrobonzyl ester for , 3-0-acetyl-l6-deacetqxy-l6a-bromofusidic acid pnitrobonzyl ester, 3-0-acetyl-l6-deacetoxy-16P5 isopropylthio-24,25-dihydrofusidic acid p-nitrobenzyl ester was prepared as. colourless crystals, melting point 113-ll6°C. · B. i6—Ρο.ηοοΐοχγ-ΐ6β- ϊ.5ορηοργ1ίΗίο-24,25-dihydrofusidic acid IQ By following the procedure of Example 1 Band substituting 3-0-acetyl-l6-deacetoxy-l6P-isopropylthi.o-24,25-dihydroftisidic acid p-nitrobenzyl ester for 3“0i“acotyl-i6-deaeetoxy-l5p-isopropylthiofusidic acid p-nitrobonzyl os Lor, 16-doacotoxy-l6p~i.sopropylj5 thio-24,25-dihydrofusidic acid was prepared, melting point 232-234°C.

Example 12 lfi-Beacetoxy-lfip-cyclohexyithiofusidic acid 3-0-Acetyl-l6-deacetoxy-l6a-bromofusidic acid p-nitro20 benzyl ester (1.43 SS 2 mmol) was added to a solution of potassium hydroxide (400 mg of 85% purity; 6.1 mmol) and cyclohexyl mercaptan (2 ml, 16 mmol) in. ethanol (lOO ml), and the resulting solution was left at room temperature for five days. Thereafter, the reaction mixture was diluted with 150 ml of ethor, washed with water (3 x 75 ml), dried and evaporated in vacuo. The residual oil, containing the crude 3-0_acetyl-l6-deacetoxyl6P-eyclohexylthiofusidic acid p-nitrobonzyl ester, was dissolved in ethanol (80 ml), and 20 ml of 2 N aqueous sodium hydroxide was added. After stirring for throe hours at 60°C, 100 ml of water was added, and the resulting dark solution was acidified with 1(1 4 N hydrochloric acid (15 fl) and extracted twice ·* with ether. The combined organic phases were washed with water (3 x 5θ ml), dried and evaporated.

The oily residue was purified by dry column chromatography on silica gel (etherspetroleum etherzacetic acidj70s 30:0.5) to give l6-deacetoxy-l6p-cyclohexylthiofusidic acid, crystallized from ether-petroleum ether, melting point: 215-220°C. Recrystallization from ethyl acetate-petroleum ether gave the analytically pure compound, melting point: 2l6-22O°C.

Examples 13-15 Following the procedure of Example 12 and substituting the mercaptans listed in table XX for cyclohexyl mercaptan, the l6P-thioethors Of 16-doacetoxyfusidic acid indicated in table XX were prepared. 4347 8 Table IX; Resulting compound Example Mercaptan R Mp (°c) c 13 2-phenyle thyl CH„CH„C,H„ 208-214 □ mercaptan y 14 n-butyl mercaptan CH2CH2CH2CH3 105-118 (dor .15 methyl mercaptanCH3 amorphous The NMR spectrum (CD^OD) of the compound of Example 15 shows signals at /= 0.89 (d, J=6, 3H), 1.00 (s, 3H), 1.03 (s. 3H), 1.38 (s, 3H), 1.62 (bs, 6lf), 2.13 (s, 3Hj SCn3), 3.03 (nr, IH; CIf-1.3), 3.67 (m, BIj CH-3), 4.03 (d, J=9, lHj CH-16), 4.26 (m, IH; CH-ll) and 5.10 (m, IH} CH-24) ppm. Tetramethylsilane was used as internal reference.

Example 16 l6-Peacetoxy-l6(3-ethylthiofusidic acid To a solution of ethyl mercaptan (2.5 ml} 34· mmol) in 'dimethylformamide {10 ml) was added sodium - 58 4347S hydride (650 mg of a 55$ suspension in oil; 15 nmol).

When the evolution of hydrogen ceased, 3-O-acetyl-l6deacetoxy-l6a-bromofusidic acid pivaloyloxymethyl ester (750 mg; 1.1 mmol) was added. After standing one hour at room temperature, the reaction mixture was diluted with ethyl acetate (50 ml) and extracted with 1 N hydrochloric acid (25 ml) and water (2x 25'ml). The organic phase was dried, filtered and evaporated in vacuo. The crude product thus obtained was purified jq by dry column chromatography on silica gel (etheri petroleum ether:acetic acid; 40s60j0.5) and afforded pure 3-0-acetyl-l6-deacetoxy-l63-ethylthiofusidic acid as a colourless gum, which was dissolved in a mixture of ethanol (20 ml) and 2 N aqueous sodium hydroxide (5 ml) and left at 75°O for two hours. Tlio reaction mixture was then acidified with 1 N hydrochloric acid (15 nil) and extracted with ethyl acetate (50 ral). The organic phase was washed twice with water (20 ml), dried, and evaporated in vacuo to give an oil, which was crystallized from ether-petroleum ether to yield l6-deacetoxy-l6p-ethylthiofusidic acid as colourless crystals, melting points 195-198°C.

Examples 17-20 16r —Thloothers of 16-deacetoxyfusi.dic acid Following the procedure of Example 16 and substituting the mei’captans listed in table X for ethyl 5 mercaptan , the l6p-thioethers of 16-deacetoxyfusidic acid shown in table X were prepared.

Table Xi Resulting compound Example Mercaptan R Mp (°C) 17 n-propyl mercaptanCH2CH2CH3 amorphous 18 t-butyl mercaptan c(ch3)3 200-203 19 phenyl mercaptanc6H5 amorphous 20 benzyl mercaptanCH2C6»5 amorphous The NMR spectrum (CD^OD) of the compound of Example 17 shows signals at <5 = 0.90 (d, 3H), 0.99 (s, 6h), 1.37 (s, 311), 1.62 and 1.66 (2 bs, 6H), 2.58 (ra, 2H; GH^S), 3.00 (m, IH; CH-13), 3.67 (m, IH; CH-3), 4.11 (d, IH; CH-16), 4.24 (m, 1H; CH-ll) and 5.12 (m, HI; CH-24) ppm.

• Tetramethylsilane was used as internal reference. - 60 43478 Tho NMR spectrum (CDCl^) of tho compound of Example 19 shows signal s at <5= 0.95 (s, 6ll), 1.10 (s, 3H), 1.35 (s, 3«), 1.60 and 1.65 (2 bs, 6ll), 3.10 (m, IH; CH-13), 3.74 (in, IH; CH-3), 4.30 (m, IH; CH-ll), 4.77 (d, IH} CH-Ιό), 5.11 (m, IH,· CH-24) and 7.0-7.4 (5H; arom. CH) ppm, Tetramethylsilane was used as internal reference.

The NMR spectrum (CDCl^) of the compound of Example 20 shows signals at J= 0.97 (s, 6η), I.36 (s, 3H), 1.62 and 1,66 (2 bs, 6n), 3'.06 (m,” IH; CH-13), 3-66 (m, IH; CH-3), 3.74 (bs, 2H; SCg2), 4.08 (d, IH; CH-16), 4.24 (m, IH; CH-ll), 5.14 (m, IH; CH-24) and 7.3 (bs, 5H; arom. CH) ppm, Tetramethylsilane was used as internal reference.

Example 21 l6“Dcacotoxy-l6(3- (11 -methyltetrazol·-5 *-ylthio)fusidic acid l6-Epideacetylfusidic acid henzoyloxymethyl ester (2.2 g; 3.4 mmol) and di(l-methyltetrazol-5-yl) di20 sulphinyl(l, 5 g; 6.5 mmol) was dissolved in dry pyridine (20 ml). The solution was cooled in an ice-bath and - 6l 43478 tributylphosphine (l.44 ml; 6 mmol) vas added. After standing for i8 hours at room temperature, water (200 ml) and ether (400 ml) was added to the reaction mixture. .

The organic phase was separated, washed twice with 1 N hydrochloric acid and twice with water, dried, and evaporated in vacuo. The residue was dissolved in methanol (50 ml)j and potassium carbonate (2.4 g; 17.5 mmol) was added. After stirring for 18 hours at room temperature, the solution'was acidified with 4 N aqueous Ιθ hydrochloric acid (8 ml), and water (200 ml) and ether (lOO ml) was added. The organic phase was separated,' washed twice with water, dried, ancf evaporated to give 1.84 g of crude product, which was purified by dry column chromatography on silica gel (ethersacetic acid; lOOsO.5) to yield 800 mg of l6-deacetoxy-l63-(l'-methyltetrazol-51-ylthio)fusidio acid as a colourless foam.

Tho nmr spectrum (CDCl^) shows signals atfi= 1,00 (s, 3Il), 1.06 (s, 31l), 1.40 (s, 311), 1.62 and 1.68 (2 bs, 6ll), 3.17 (in, 1M; CH-13), 3.75 (m, IH; CH-3), 3.87 (s, 3H; l‘“CS3), lf.37 (m, 111, CH-ll) and 5.42 (m, III; CH-16) ppm.

Tetramethylsilane was used as internal reference.

Example 22 l6-r)oacetoxy-l6P-(2', S'-dichloro-phcny l.thio)fusldi π acid A solution of 3-O-*acetyl-l6-epideacetylfusidic acid mothoxymethyl ester (4?0 mg; Ο.87 mmol) and di(2,5-di- 62 43478 chloroplienyl) disulphide (l.07 g; 3-75 mmol) in dry pyridine (4 ml) was cooled to 0°C, and tributylphosphine (0.72 ml; 3.0 mmol) was added. The resulting solution was left at 5°C for 3 days and then diluted with other (100 ml), washed with 4 N hydrochloric acid, N sodium hydroxide and water, dried, and evaporated in vacuo. The residue was dissolved in a mixture of ethanol (20 ml) and 2 N aqueous sodium hydroxide (8 ml), and kept at 60°C for 1 hour. The reaction mixture was then acidified with 4 N hydrochloric acid (5 ml), and ether (100 ml) and water (200 ml) was added. The organic * phase was separated, washed twice with water, dried and evaporated in vacuo. The residue was crystallized from ether-petroleum ether to give l6-deacetoxy-l6P-(2’,5’-dichlorophenylthio)fusidic acid, melting point l6l-l64°C.

Example 23 l6-Doacetoxy-l66-(2'-azidoethylthio)fusidic acid A, 3-Q-Acetyl-l6-doacetoxy-l6P-(2'-bromoethylthio )fusidic acid p-nitrobenzyl ester To a solution of 3-0-acetyl-l6-deacetoxy-l6p-(2'-hydroxyethylthio)fusidic acid p-nitrobenzyl ester (l g; 1.4 mmol) in 50 ml of dimethylformamide was added g of phenyl N,N-dirnothylformimidate bromide. After standing at room temperature for 18 hours, the reaction mixtuie was diluted with ether (50 ml), washed with - 63 43473 ι N sodium’ hydroxide (20 ml) and water (3 x 50 ml), dried, and evaporated in vacuo. Addition of etherpetroleum ether caused the residue to crystallize.

The product was filtered off, washed with petroleum ether, and dried to yield 800 mg of 3-O-a-cetyl-l6» -deacetoxy-l63-(2,-hromoethylthio)fusidic acid pnitrobenzyl ester, melting point l48-15O°C.

B. 16-Doacetoxy-l6j3- (2'-azidoethylthio)fusidlc acid The 2 *-bromoethylthioether prepared above was disj0 solved iii 25 ml of dimethylformamide, lithium azide (400 mg; 8.2 mmol) was added, and the reaction mixture was left at 20°C for 24 hours. 100 ml of ether was then added, and the resulting solution was washed with water (4 x 50 ml), dried, and evaporated in vacuo. The residue was dissolved in a mixture of ethanol (50 ml) and 2 N aqueous sodium hydroxide and after being left at 60°C for 3 hours, the solution was acidified with 8 ml of 4 N hydrochloric acid, and water (100 ml) and ether (100 ml) was added. The organic phase was separated, 2Q washed with water (4 x 50 ml), dried, and evaporated in vacuo. Addition of ether and petroleum ether to the residue caused 16-dcacetoxy-l6p-(2'-azidoethylthio)fusidic acid to precipitate as colourless crystals, which were filtered off, washed with petroleum ether, and ,5 dried to yield l4o mg, melting point 173-179°C. - 64 43478 Example ·?4 16-I>oacotoxy-1 hf'- (21 -methoxynthyl thio)fusidie acid sodIum snIt To a solution of 3-0-acotyl-l6-doacotoxy-l6p~(2'5 -bromoethylthio)fusidie acid p-nitrobenzyl ester (see Example 23 A for the preparation of this compound) (l g; 1.3 mmol) in methanol (50 ml) was added silver carbonate (l g; 3.6 mmol), and the mixturo was stirred at room temperature for 16 hours. Tho insoluble material was filtered off and washed with methanol (lO ml). The combined filtrate and washing were evaporated in vacuo. and the residue was dissolved in a mixture of ethanol (100 ml) and 2 N sodium hydroxide (20 ml). After stirring for 3 hours at 60°C, the dark solution was acidified with 4 N hydrochloric acid (15 ml), a’-d water (200 mi) and other (.200 ml) was added.

Tho organic phase was separated, washed twice with water, dried and evaporated in vacuo. The residue was purified by dry column chromatography on silica gel 2θ (ethersacetic acid; 100:0.5) to give the desired product as a colourless foam, which was converted into a crystalline sodium salt by dissolving in methanol (25 ml), titrating with 2 N aqueous sodium hydroxide, evaporating, and adding acetone. Tho crystals wore filtered off, washed with acetone, and dried to yield l6-deacetoxy-Ι6β~(2*-methoxyethylthio)fusidic acid sodium salt. - 65 43478 The NMR spectrum (CD OH·) shows signals at & = 1.00 (s,6h), 1.36 (s, 3Π), 1.62 (bs, 6h) 2.78 (211; CHgS), 3.51 (211; CHgO), 3.68 (m, IH; CH-3), 4.10 (d, IH; CH-16), 4.21 (ιιι, HI; CH-ll) and 5.11 (in, IH; CH-24) ppm. To trainethyls i.J ano was used as internal reference.

Example 25 l6-l)eacetoxy-168-(21-isopropylthioethylthio)fusidic acid To a solution of potassium hydroxide (5°0 mg; 9 mmol) and isopropyl mercaptan (ΐζ5 ml; l6 mmol) in ethanol (50 ml) was added 3-0-acetyl-l6-deacetoxy-l6j3-(2·-bromoethylthio)fusidic acid p-nitrobenzyl ester (see Example 23A for the preparation of this compound) (l g; 1.3 mmol), and the mixture was stirred for 16 hours at room temperature. Water (lOO ml) and ethor (75 ml) was added, the organic phase was separated, washed with 2 N sodium hydroxide (2 X 25 ml) and water to (2 x 25 ml), dried, and evaporated in vacuo. The residue was dissolved in a mixture of ethanol .(lOO ml) and 2 N sodium hydroxide (20 ml), and the solution stirred for 3 hours at 60°C. 4 N hydrochloric acid (15 ml), water (250 ml) and ether (100 ml) was added, the organic phase was separated, washed with water (2 x 50 ml), dried, and evaporated in vacuo. The desired product was - 66 43478 isolated from the residue by dry column chromatography (ethor:petroleum etliersacetic acid, 7θί3θ!°·5) to yiel'd 400 mg of l6-deacetoxy-l6p-(21-isopropylthioethylthio)fusidlc acid as a colourless foam.

Tho nmr spectrum (CDCl^) shows signals at 6 = Ο.96 (bs, 611), 1.22 (d, J=7, 61l), 1.33 (s, 3»). 1.58 and 1.67 (2 bs, 6h), 2.73 (bs, ΑϊΓ; SCHgCHgS), 2.91 (m, 1Η; S-CH (CH3)2), 3.01 (m, 1H, CH-13), 3.71 (m, IH; CH-3), 4.21 (m, IH; CH-16), 4.28 (m, IH; CH-ll) and 5*08 (m, IH, CH-24) ppm. Tetramethylsilane was used as internal reference.

Example 26-28 By following the procedure described in Example 25 and substituting the mercaptans listed in table XI for isopropyl mercaptan, the compounds indicated in table XI were prepared Table XI: Η°/γ-~χ4 fjEHI ΙΓ COOH SCHgCHgSR Example Mercaptan R Mp (°C) 26 ethyl mercaptan ch2ch3 149-152 27 t-butyl mercaptan c(ch3)3 134-135 28 cyclohexyl mercaptan cyclohexyl amorphous - 67 43478 Tho NMR spectrum of the compound of Example 28 (CDCl^) shows signals at ?CH?S), 3.07 ' (in, IH; CH-13), 3.76 (m, IH; CH-3), 4.26 (d, IH; CH-16), 4.35 (m, IH; Cn-11) and 5.12 (m, III; CH-24) ppm, Tetramethylsilane was used as internal reference.

Example 29 l6-Deacotoxy-l6|3-(2'-phenylthioethylthip)fusidic acid IQ To an ice-cooled solution of 3-0-acetyl-l6-deacetoxy -Ι6β-(2 ’-liytlroxyothy.'l t;h lo) fusidic acid p-ni trobonzyl ester (l g; 1.4 mmol.) and diphenyl disulphide (l g; 4.6) mmol) in dry pyridine (7 ml) was added tributylphosphine (2 ml; 8,4 mmol), and the mixture was left at 5°C for 16 hours. Etiier (100 ml) was added, and the resulting solution was washed with 4 N hydrochloric acid (2 x 25 ml), 2 N sodium hydroxide (2 x 25 ml) and water (2 x 25 ml), dried, and evaporated in vacuo. The residue was dissolved in a mixture of ethanol (90 ml) and 2 N sodium hydroxide (20 ml). After stirring for hours at 60°C, 4 N hydrochloric acid (15 ml), water (200 ml) and ether (lOO ml) was added. Tlie organic phase was separated, washed with water (2 x 20 ml), and evaporated in vacuo. The residue was purified by dry column chromatography on silica gel (ethyl acetate;cyclohexane; l:l) to yield 6 30 mg of l6deacetoxy-l6p-(2'-phenyltbioethylthio)fusidic acid ( as a colourless foam.

The nmr spectrum (CDCl^) shows signals at <$ = 0.98 (bs, 611), 1.35 (s, 3H), 1.61 and 1.67 (2 bs, 6h), 3.78 (m, III; CH-3), 4.25 (m, IH; CH-16), A.3A (m, IH; CH-ll), 5.11 (m, IH; CH-24) and 7.1-7-5 (m, 5H; arom.

CH) ppm. Tetramethylsilane was used as internal 2θ reference.

Example 30 · * Sodium h;i 11 of l6-deacetoxy-l6p-(21-methylthioethylthio)fus id je acid To an ice-cooled solution of 3-O-acetyl-l615 deaoetoxy-l6p-(2'-liydroxyethylthio)fusidic acid pnitrobenzyl ester (l g; 1.4 mmol) in 1 ml of dimethyl disulphide (v-\10 mmol) was added tributylphosphine (2 mt; 8.4 mmol), and tho mixture was left at 20°C for 3 days. Ether (100 ml) was then added, and the resulting solution was washed with 2 N. sodium hydroxide (25 ml) and water (2 x 25 ml), dried, and evaporated In vacuo. Tho residue was dissolved in a mixture of ethanol (4o ml) and 2 N sodium hydroxide (10 ml).

After stirring for 3 hours at 60°0, 4 N hydrochloric 2$ acid (lO ini), water (200 ml), and ether (100 ml) was added. The organic phase was separated, washed with - 69 43478 water (3 χ 20 ml), and evaporated in vacuo. The residue was purified by dry column chromatography on sil ica gel (etheripetroleum othersacetic acid; 70:30:0.5) to yield 410 mg of l6-deacetoxy-l6p-(2'-methylthioethylthio)fusidic acid as a colourless oil. The crystalline sodium salt was prepared by dissolving this oil in methanol (lO ml), titrating with 2 N aqueous sodium hydroxide using phenolphthalein as indicator, evaporating in vacuo. and adding acetone, The crystals were filtered off and washed with acetone and ether to yield the pure sodium salt of 16-deacetoxyΙ6β-(2’-methylthioethylthio)fusidic acid.

The NMR spectrum (CD^OD) shows signals at ό = 0.98 (s, 6h), 1.36 (s, 3H), 1.62 (bs, 6H), 2.10 (s, 3H; SCH3), 2.77 (bs, 4ll; SCH^HgS), 3.00 (m, IH; CH-13), 3.66 (in, III; CH-3), *1.11 (d, 111; CH-16), 4.23 (m, IH; ‘CH-1J.) and 5.13 (», 111; CH-24) ppm. Tobraniothylsilanc was used as internal reference.

Example 31 l6-Deacotoxy-l6P-(21-fluoroethylthio)fusidic acid A. 3-QwForinyl-X6-deacotoxy-160-(21-bromoethylthio )fusidic acid benzoyloxymethyl ester - 70 43478 16-Deacetoxy-ΐόβ-(2'-hydroxyethyithio)fusidic « · acid (53.4 n,g; 1 mmol) was dissolved in methanol (10 ml) and converted into its sodium salt by titration with 2 N sodium hydroxide using phenolphthalein as indicator.

After evaporation in vacuo, the resulting amorphous sodium salt was dissolved in dimethylformamide (7.5 .ml), chloromethyl benzoate (0.16 ml; 1 mmol) was added, and tho mixture was stirred at room temperature for 48 hours Water (50 ml) was added, and the mixture was extracted with ether (lOO ml). The organic pjiase was separated, washed with water (4 x 100 ml), dried and evaporated in vacuo to yield 16-deacetoxy-l6p-(2*-hydroxyethyitliio)fusidic acid benzoyloxymethyl ester as an amorphous product. This was dissolved in dimethylformamide (10 ml), phenyl N,N-dimethylformimidato bromide (l.5 g; ca. 6.5 mmol) was added with stirring, and the solution was kept at 20°C for 24 hours. Water (50 ml) and ether (50 ml) was added, and the organic phase was washed with 2 N sodium hydroxide (2 x 25 ml) and water (2 x 25 ml), dried, and evaporated in vacuo to give 3-0-formyl-l6-doacetoxy-l6p-(2'-bromoethylthio)fusidic acid benzoyloxymethyl ester as an amorphous produc t. Β. 16-1)eacn toxy-16)3-(21 -fluoroothyl thio) fusid lc acid The 2'-broinoo thyl thioether prepared above was dis The filtrate was evaporated in vacuo. the' residue was dissolved in methanol (lO ml), and potassium carbonate (350 ing; 2.5 mmol) was added. After stirring for 30 minutes at room temperature, water (lOO ml), 4 N hydrobhloric acid (5 ml) and ether (lOO'inl) was added, and the organic phase was separated, washed twice with water, dried, and evaporated to yield an amorphous product, which was purified by dry column chromatography (ether: acetic acid; 100:0.5) 1° yield pure 16-deacetoxyl6p-(2’-fluoroethylthio)fusidic acid, crystallized from ether-petroleum ether, melting point 157-159°C.

' Example 32 1l-Dehydro-l6-deacetoxy-l6g-isopropylthiofusidic acid.

A. 3-O-Acetyl-ll-dehydro-l6-deacetoxy-l6g-isopropylthiofusidic acid p-nitrobenzyl. ester Pyridinium chlorochromate (l.07 g, 5 mmol) was suspended in methylene chloride (3θ ml) hy stirring, While 3-Q-aeotyl-l6-doacetoxy-l6f3-isopropylthiofusidic - 72 43478 acid p-ni, trebenzy I ester (l.5 G« 2.1 mmol) was rapidly added. After stirring for an additional hour, the , suspension was diluted with ether (100 ml), the solvent was decanted, and the black solid was washed twice ς with ether. Filtration and evaporation of the combined organic extracts yielded an oily residue, which crystallized from ether-petroleum ether. The colourless crystals thus obtained were collected, washed with petroleum ether, and dried to afford 880 mg of the desired product, melting point 120-122°C. z B. ll-Dchydro-l6-deacetoxy-l6g-isopropylthiofusidic acid • The p-nitrobenzyl ester prepared above was dissolved in a mixture of ethanol (20 ml) and 2 N aqueous sodium hydroxide (5 ml) and heated to 60°C for 3 hours. Then, 4 N hydrochloric acid (3 ml), watex- (lOO ml) and ether (100 ml) was added with stirring. The organic phase was separated, washed twice with water (25 ml), dried, and evaporated, in vacuo. Tho resulting oily residue was purified by dry column chromatography on silica gel (cyclohexane:ethyl acetate, 7:3) to yield 380 mg of 11- dehydro-16-deacetoxy-l6B-isopropylthiofusidic acid, melting point l67-l69°C (crystallized from etherpetroleum ether). - 73 43478 Example 33 11-Ρ6Ηγά·Γο-ΐ6-άβαοβΕοχν-ΐ6β-Ϊ8ορΓθρν1~ίΗϊο-24·32ζ!.-άϊΗγdrofusidic acid By following the procedure of Example 32 but 5 substituting 3-0-acotyl-l6-doacetoxy-l63-isopropylthio-24,25-dihydrofusi.dic acid p-nitrobenzyl ester for 3-0-acety] -l6-deaeetoxy-l6fi—isopi-opyltiiiofusidic acid p-nitrobcnzyl ester, Il-dehydro-l6-deacefcoxy~l6β isopropylthio-24,25-dihydrofusidic acid was prepared as colourless crystals, melting point 189-191°C.

Example 34 3-Dehydro-l6-deacetoxy-l6g-isopropylthiofu.sidic acid To a solution of l6-deacetoxy-l6P-isopropylthiofusidic acid acetoxymethyl ester (2.0 g, 3.3 mmol) in 15 ml of dimethyl sulphoxide was added dicyclohexyl• carbodiimido (3.10 g, 15 mmol) and orthophosphoric acid (l60 mg, 2 mmol), and the mixture was left with stirring at room temperature for 24 hours. .A solution of oxalic acid (3 g) in methanol (20 ml) was then added to destroy excess of carbodiimide, and stirring was continued for 3θ minutes. Ethyl acetate (150 ml) was then added, and the resulting solution was washed with saturated aqueous sodium bicarhonate - 74 43478 (2 χ 50 ml) and water (50 ml), dried, and evaporated to yield J .9 g »f ail oily residue. This was dissolved in methanol (4o ml) and potassium carbonate » (i.2 g) was added. After stirring for one hour, the methanol was evaporated in vacuo, and ether (100 ml) and 4 N hydrochloric acid (50 ml) were added to the residue. Tiie organic phase was washed with water (2 x 50 mi), dried, and evaporated in vacuo. The oily residue was purified by dry column chromnto10 graphy on silica gel (cyclohexane:ethyl acetate, 7:3) to yield 3-dehydro-16-deacetoxy-l6 g-isopropylthiofusidic acid as colourless crystals,, collected from ether, melting point 200-203°C.

Example 35 l6-neaeetoxy-l6j3—isopropylsulphinylfuBidic acid Sodium metaperiodate (6 g; 28 mmol) in 500 ml of water was added to a solution of 16-deacetoxy-l6p-isopropylthiofusidic acid (10.0 g; 18.3 mmol) in a mixture of methanol (200 ml) and 2 X aqueous sodium hydroxide (lO ml). After standing for 1.5 hours, the resulting solution was acidified with 4 N aqueous hydrochloric acid (7·5 ml), causing a crystalline product to precipitate. The crystals were filtered off, washed with water (50 ml), and dried to yield 10.0 g of the desired product, mp. 158-139°C. The crystals thus obA . tained were transformed into another crystal modifica- 75 43478 t j on by treating witli boiling ethyl acetate (400 ml).

After cooling to 0°C, the product was filtered off washed with ether (50 nil), and dried to afford 9·θ^ g, of pure l6-deacetoxy-l6P-isopropylsulphinylfu.sidic acid, melting point 179-l8l°C.

Example 36-41 By following the procedure desci-ibed in Example 35 but substituting the 16J3-thioethers of l6-deacetoxyfusidic acid listed in table XXX for l6-deacetoxy-l6P-isoIQ propyl tliiof usidic acid, the sulphoxides indicated in table XIX were prepared Exam- ple thioethor of 16-deacctoxy fusidic acid R Mp (°C) 36 Ιδβ-inethylthioether ch3 151-156 37 ΐδβ-ethylthioether ch2ch3 158-162.5 38 ΐ6β-t-butylthioether c(ch3)3 164-167 39 Ι6β-(2'-hydroxyethylthio)ether CHjjCHgOH 163-I6S 4o 16β- (2 '-azidoe tliyl thio) other ch2ch2x3 l4l-l4? 4l Ιόβ-phonylthioeI herc6fl5 amorphous - 76 43478 Example 42 16-Doacetnxy-1 6 i sopropyl snlphinyl-24, 25-dihydrofusidlc acid By following the procedure described in Example 35 but substituting i.6-deacetoxy-l6p-isopropylthio24,25-dihydr’ofusidic acid for l6-deacetoxy-l63isopropylthiofusidic acid, l6-deaoetexy-l6p-isopropylsulphinyl-24,25-dihydrofusidic acid was propared as colourless crystals, melting point 184~186°C.

✓ Example 43 -e-— l6-Doacotoxy-3-dehydro-l6B-isopropylsulphinylfusidic acid By following the procedure described in Example 35 but substituting l6-deacetoxy-3-dehydro-l6(?-isopropyl15 thiofusidic acid for l6-deacotoxy-l6(3-isopropylthiofusidic acid, l6-deacetoxy-3-dehydro-l6g-isopropylsulphinylfusidic acid was prepared as colourless crystals, melting point 158-l6l°C.

Example hh ll-Dehydro-l6-deacetoxy-l6 R-isopropylsulphinylfusidic acid To a solution of l6-deacetoxy-l6p-isopropylsuphinylfusidic acid (l.l g; 2 mmol) in 5 ml of pyridine was - 77 43478 added acetic anhydride (o,8 ml; 8.5 mmol). After standing for 48 hours at room temperature, 1 ml of water was added to the solution, which after an additional hour was diluted with 50 ml of ethyl acetate, washed twice with 4 N hydrochloric acid and twice with water, dried, and evaporated to give 940 mg of crystalline 3-0-acetyl~l6-deacetoxy-l6p-isopropylsulphinylfusid±c acid collected from ether, melting point 176-178°C.

To a suspension of 770 mg of this product in acetezne 10 (lOO ml) was added Jones reagent (0.7S ml). After standing for 10 minutes at room temperature, water (lOO ml) was added to the reaction mixture, and the resulting solution was concentrated in vacuo to 125 ml causing 3-0-acetyl-ll-dehydro-l6-deacetoxy-l68-isopropylj5 sulfinylfusidic acid to precipitate as colourless crj'stals, which were filtered off, washed with water and dried to yield 570 mg; molting point 151-l6O°C. 400 mg of this product were dissolved in a mixture of ethanol (20 ml) and 2 X aqueous sodium hydroxide (2 ml), and left at room temperature for 6 days. 4 N aqueous hydrochloric acid (2 ml) was then added with stirring to precipitate the desired product as colourless crystals, which were collected, washed with water (15 ml), and dried to afford 230 mg, melting point 174-178°C. - 78 43478 RecrystaI I 1zation from ethyl acetate gave tho pure 11 “dcliydro-l6-deacetoxy-l6P-isopropylsulphinylfusidic acid, melting point 181-183°C.

Example 45 .3,11-Bisdehydro-16-deacetoxy-l6 R-isopropvlsulphlnylfusidfc acid To a solution of l6-deacetoxy-l6p-isopropy.l.thiofusidic acid (500 mg; 0.94 mmol) j.p 100 ml of acetone was added 1.6 ml of Jones reagent. After standing at Ιθ room temperature for 10 minutes, 100 ml of water was added with stirring to the reaction mixture. The white precipitate which formed was filtered off, washed with water and dried to give 450 mg of a mixture of 3,11-bisdehydro-lή-deacetoxy-l6β-isopropylthiofusidic acid and the desired product.

The crystals were dissolved in hot ether (20 tni) and upon cooling to 0°C pure 3»H-bisdehydro-l6-deacetoxy-l6p-isopropylsulphinylfusidic acid precipitated. The crystals were filtered off, washed with cold ether and dried to give 6o mg, molting point 154-102°C.

Example 46 l6-Deacctoxy—163-ethoxyfusldic acid ' Silver carbonate (16.55 Si 60 mmol) was added to a suspension of 3-0-acetyl-l6-deacetoxy-l6a-bromofusidic 5 ' acid phenacyl ester (20.94 g; 30 mmol) in ethanol (300 ml) and, after being protected from light, the mixture was stirred at room temperature for 18 hours. Insoluble material was filtered off and washed with ethanol (2 x 30 ml), To the combined filtrate and washings was added 5 N aqueous sodium hydroxide (l20 ml), and z the mixture was refluxed fo'r two hours. After cooling to room temperature, the major part of ethanol was removed in vacuo, and to the residue was added ethyl acetate (l50 ml) and water (lOO ml). The stirred mixture was acidified with 4 N hydrochloric acid, the organic phase was separated, and the aqueous phase reextracted / with ethyl acetate (50 ml). The combined organic extracts were washed with water, dried, and evaporated in vacuo to yield an oily residue, which crystallized 2o from diisopropyl ether. The colourless crystals thus obtained were collected, washed with diisopropyl, ether, and dried to afford 5.42 g of l6-deacetoxy-l6p-ethoxyfusidic acid, melting point: 169-171°C. After work-up of the mother liquor, a further 2.20 g of the desired compound, melting point: l68-170°C, was obtained.

Two recrystallizations from methanol-diisopropyl ether gave the analytically pure compound, melting point: 177-178°C.

Examples 47-49 l6-l)oacetoxy-l68-alkyloxyfusldlc acids By substituting the alcohols listed in table XIII for the ethanol in the procedure of Example 46, the l6~ 20 deacetoxy—ΐ6β—alltyloxyfusidic acids indicated in table XIII were obtained.

Table XTII: Resulting compounjl Example Alcohol R Mp (°C) 47 Methanol ch3 175-176 48 2,2,2-Trifluoroethanol ch2cf3 202-203 49 Hexanol-(l) ch2(ch2)4ch3 amorphous Example 50 . l6-Deacotoxy--.l6i3--(2l-fluoroethoxy).fusidic acid To a solution of 3~0—formyl—16—deacetoxy—16a— bromofusidic acid benzoyloxymethyl ester (8.75 gj 12.5 mmol) in 2-fluoroethanol (25 ml) was added silver carbonate (6.89 g; 25 mmol), and the mixture was stirred at room temperature and in the absence of light for l6 hours. The insoluble material was filtered off, washed twice with ether, and the combined filtrate and washings were evaporated to dryness in vacuo. The residual * oil, containing the crude J-0-forniyl-l6-deacetoxyl6p-(2’-fluoroethoxy)fusidic acid benzoyloxymethyl ester, was dissolved in methanol (85 ml), potassium carbonate (3.46 g; 25 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. The major part of the solvent was removed by evaporation in vacuo. and to the residue was added water (100 ml) and ether (lOO ml). After acidification of the stirred mixture with 4 N hydroclilox’ic acid, the organic layer was 2q separated, the aqueous layer reoxtracted with ether (50 ml), and the combined organic phases were washed with water until neutral. In order to separate the desired acid derivative from l6-deacetylfusidio acid lactone, boing formed as a by-product, tho ethereal - 82 43478 solution obtained above was extracted with 0.5 N sodium hydroxide (3 x 50 ml) and washed with water (3 x 25 ml). To the combined aqueous phases and washings was added ether (lOO ml), and the stirred mixture vas acidified with 4 N hydrochloric acid.

After separation of the·organic layer, the aqueous layer was extracted with ether (50 ml), and the combined organic extracts were washed with water until neutral, dried, and evaporated in vacuo. The rejo suiting amorphous residue was dissolved in diisopropyl ether (30 ml), and on scratching a crystalline product precipitated. After being kept in the refrigerator overnight, the crystals were filtered off, washed with diisopropyl ether, and dried to afford 2.32 g of l6-deacetoxy-l6P-(2'-fluoroethoxy)fusidic acid, melting point: 158-l60°C. From the mother liquor a further 0.48 g of tho desired compound, melting point: 155-159°C, vas obtained. Two rccrystallizations from inethanoldiisopropyl ether gave the analytically pure product, melting point: l62-l6'_)°C. - 83 43478 Examples 51-52 l6-Heaeotoxy-l6p-aTkyloxyfus.tdic acids , Following the procedure of Example 5θ, but substituting the alcohols listed in table XIV for the 25 fluoroethanol, the l6-deacetoxy-l6p-alkyloxyfus.i<3ic acids indicated in table XIV were obtained.

Table XIV: Resulting compouri Example Alcohol Mp (°C) 2-Ac e toxye tlianol 1,3-Difluoropropanol-(2) CH2CII20H CH(CH2P)2 179-182 169-171 Examples 53-62 Ιόβ-Ethers of 16-deacetoxyfusidic acid Following the procedure of Example 50 , but substituting l6-deacetoxy-l6a-bromofusidic acid acetoxymethyl ester for the 3-0“formyl-l6-deaeetoxy-l6££-bromofusidio acid benzoyloxymethyl ester and the alcohols - 84 434 78 listed in table XV for the 2-f luoroetlianol, the Ι6βcthers of l6-deacetoxyfusidic acid indicated in table XV were obtained.

Table XV; z . COOH H0\ z * Resulting compound Example Alcohol R Mp (°C) 53 Isopropanol ch(cii3)2 189-190 54 tert.Butanol c(ch3)3 179-180 55 2,2-Dichloroethanol CHgCHClg 181-182 56 2,2,2-Trichloroethanol ch2cci3 212-213 57 1,3-Eifluoropropanol-(2) cii(ch2f)2 169-171 58 1,3-Diacetoxypropan- ol-(2) ch(ch2oh)2 amorphous 59 Allyl alcohol ch2ch=ch2 154-156 60 2-Butenol-(l) CH2CH=CHCII3 128-135(6 61 2-Propynol-(l) ch2c=cii 134-136 62 Cyclopentanol cyclopentyl 188-189 Example 6.3 l6-Deacetoxy-l6P-ethoxv-24,25-dihvdrofusidic acid A, 3-0-Acoty1.-1.6-epideacetyl-24, 25-dlhydrofusidic acid pivaloyloxymethyl ester To a solution of 3-O-acctyl-l6-epideacetyl-24,25dihydrofusidic acid (31.12 g; 60 mmol) in dimethylformamide (25O ml) was added triethylamine (ll.92 ml; mmol) and, after stirring for 15 minutes, chloromethyl pivalato (17.76 ml; 120 mmol). After stirring for 20 hours at room temperature, the mixture was diluted with ethyl acetate (750 ml) and washed thoroughly with water (4 χ 25Ο ml, 2 x 50 ml) to remove unreacted starting, material and the greater part of dimethylformamide.

The organic phase was dried and evaporated in vacuo to yield 42 g of an oily residue. The residue was dissolved in ether (50 ml), petroleum ether (200 ml) was added, and the mixture was stirred for two hours. The crystalline precipitate thus obtained was filtered off, washed with ether:petroleum ether 1:4. The combined filtrate and washings were evaporated to dryness in vacuo to give 36 g of crude 3-O-acetyl-l6-epideacetyl-24,25-dihydrofusidic acid pivaloyloxymethyl ester as a foam which failed to crystallize. - 86 43478 I). · 3-0-Acety L-16-deace toxy-l6o:-nic thane sul fortyl oxy24,25-dihydrofusidic acid pivaloyloxymethyl ester To a stirred solution of crude 3-O-acetyl-l65 epideacetyl-24,25-dihydrofusidic acid pivaloyloxymethyl ester (30 g; containing ~ 45 mmol of pure compound) in a mixture of methylene chloride (75 ml) arid pyridine (75 ml) was added dropwise at -20°C a solution of methanesulphonyl chloride (13.8 ml;'* l80 mmol) in methylene chloride (25 ml). After the addition was finished (ca, 15 minutes), the mixture was stirred at -15°c for 1,5 hours and then kept in the refrigerator overnight. Ice (ca. 15 g) vas added and, after stirring for 0.5 hour, tho mixture was poured into a mixture of other (250 ml) and water (lOO ml) arid shaken vigorously The organic layer was separated and the aqueous phase roextractod with ether (100 ml). The combined organic phases were washed with water, 4 N hydrochloric acid (to remove pyridine), saturated aqueous sodium chloride, 0.5 M aqueous sodium bicarbonate, and once more saturated aqueous sodium chloride, dried, and evaporated 1n va cun to yiold 28.5 g of crude 3-0-acotyl-l6-deacetoxy-l6amethanesulphonyloxy-24,25-dihydrofusidic acid pivaloyl87 43478 oxyme l.hy 1 ester as a yellowish foam which failed to crystallize. Tho relatively unstable product was used for the next stop without further purification; IK (KBr): 1170 and 1365 cm-1.

G. 3-0-Aco tyl-16-cleacotoxy-l6P-ethoxy-24,Sg-dihydrofusidic acid pivaloyloxymethyl ester A solution of crude ,3-0-acetyl-l6-deacetoxy-l6amethanesulphonyloxy-24·,25-dihydrofusidic acid pivaloyloxymethyl ester (2.6 g) in ethanol (25 ml) was stirred jq at 6O-65°C for two hours. Water /100 ml) was added, and the mixture was extracted with ethyl acetate (2 x 25.ml). The combined organic extracts were washed with water, dried, and evaporated in vacuo to give I.98 g of a yellowish gum. The residue was purified by dry column chromatography on silica· gel (cyclohexane ethyl acetate 85:15) to yield 0.72 g of 3-0-acetyl-l6-deacetoxy-l6pethoxy-24,25-dihydrofusidic acid pivaloyloxymethyl ester as a colourless foam.

D, l6-Dcacetoxy-l6p-ethoxy-24,25-dihydrofusidlc acid To a solution of the Ιδβ-ethoxy ester described above in ethanol (10 ml) was added 5 N aqueous sodium hydroxide (2 ml), and the mixture was kept at room temperature overnight. After addition of water {50 ml), the mixture was acidified with 4 N hydrochloric acid and extracted with ethyl acetate (2 x 25 ml). The ’ combined organic extracts were washed with water, driod, and evaporated in vacuo to leave 0.52 g of an amorphous product which crystallized from ether. The crystals were filtered off, washed with ether and dried to afford 0.26 g of l6-deacetoxy-l6p-etlioxy-24,25dihydrofusidic acid, melting point: 189-Ι91°θ· Two recrystallizations from ether gave the analytical sample, melting point: 192-193°C.

Example 64 l6-Deacetoxy-l6p-mottioxy-24,25-dihydrofusldic acid A. 3-Q-Aeotyl-J 6-doaceloxy-l6p-niothoxy-24,25-dihydro15 fusi.riic acid pivaloyloxymethyl ester Following the procedure of Example 63 A—C, but substituting methanol for the ethanol, 3-0—acetyl—16-deacetoxy-l6fi-methoxy-24,25-dihydrofusidic acid pivaloyloxymethyl ester was obtained.

B· l6-Deacetoxy-l6j3-niethoxy-24,25-dihydrofusidic acid By substituting 3-0-acetyl-l6-deacetoxy-l6p-methoxy24,25-dihydrofusidic acid pivaloyloxymethyl ester for the corresponding Ιδβ-ethoxy derivative in the procedure of Example 63 D, l6-deacetoxy-l6|3-methoxy-24,25-dihydroor fusidlc acid, melting point 152-154°C, was obtained.

- KO Example 65 l6-Deacetoxy-l6B-propyloxyfusidic acid A. 3-0-Acetyl-l 6-deaoetoxy-l6c<:-methanesulphonyloxyfusidic acid piva1oyloxymethyl. ester A solution of methanesulphonyl chloride (4.6 ml; ''•w 60 mmol) in methylene chloride (10 ml) was added dropwise at -20°C to a stirred solution of crude 3-0acetyl-16-epideacetylfusidic acid pivaloyloxymethyl ester (10 g; containing 15 mmol of pure compound) ;0 in a mixture of methylene chloride (25 ml) and pyridine (25 ml), placed in a 3“neched 250 ml-flask equipped with a thermometer, a dropping funnel, and a drying tube. After the addition was finished, the mixture was stirred at -15°C for 1.5 hours and then kept in the refrigerator overnight. Ice (ca. 5 g) was added, :. and after stirring for 0.5 hour, the mixture was poured into water (50 ml) and extracted with ether (2 x 50 ml). The combined organic phases were washed with water, 4 N hydrochloric acid (ta remove pyridine), saturated aqueous jq sodium chloride, 0.5 M aqueous sodium bicarbonate, and again saturated aqueous sodium chloride, dried, and evaporated in vacuo to afford 10.6 g of crude 3-0-acetyll6-deacotoxy-l6a-methanesulphonyloxyfusidic acid pivaloyl- 90 43478 oxymethyl psler as a yellowish amorphous product. Πιο unstable compound was used for t-lit? next step without further purification.

IH (KBr): 1170 and 1355 cm-1.

By using tiie method described above, but substituting p-toluenesulphonyl chloride for the methanesulphonyl chloride, the corresponding l6a-p-toluenesulphonyloxy derivative was prepared.

B, l6-Deacotoxy-l63-propyloxyfustdic acid To a solution of crude 3-0-acetyl-l6-deacetoxy-l6ftmethanesulphonyloxyfusidic acid pivaloyloxymethyl ester (1.42 g; c 2 mmol) in propanol-(l) (10 Ml) was added triethylamine (θ,28 ml; 2 mmol), and the mixture was stirred at room temperature for 42 hours. After dilution with ethyl acetate (40 ml), the mixture was washed with water, diluted hydrochloric acid, and water, dried, and evaporated in vacuo to give 1.28 g of an amorphous product. This residue was purified by dry column chromatography on silica gel (petroleum ether; ethyl acetate; 85:15) to yield Ο.36 g of 3-°-acetyl-l6deacotoxy-l6p-propyloxyfusidic acid pivaloyloxymethyl ester as a colourless foam. The above ester was hydrolyzed by refluxing its solution in ethanol (5 ml) with 5 N aqueous sodium hydroxide (l ml) for 2 hours. After a similar work-up procedure as described in Example 63 D, crystalline l6-deacetoxy-l6p-propyloxyfusidic acid, melting points 176-177°C, was obtained. - 0! Examples 66-70 16-Beaee toxy-1.6P-al kyl oxy fusidic ac i ds By substituting the alcohols lasted in table XVI for kilo propanol-(l) in tho procedure of Example 65, the l6-deacetoxy-l6p-alkyloxyfusidic acids indicated in table XVI were obtained.

Table XVI; Resulting compound Example Alcohol R Mp (°C) 66 Butanol-(l) οη2οη2οη2οη3 167-169 ' 67 iso-3utanol CH2CH(CH )2 189-190 68 2-Me thoxy e thano1 ch2ch2och3 163-165 69 2-Chloroethanol ch2ch2ci 158-159 70 Benzyl alcoholCH2C6H5 113-119 _ 92 _ Example 71 16-Doaeetoxy- l6p - ( 2 1 ~-iz j doe tlloxy) fusidic ac i d A. 3-0-Acety]-l6-deacetoxy-l6p-(21-hydroxyethyloxy)fusidic acid phenacyl ester To a solution of 3-0-acetyl-l6-deacetoxy-l6(X-bromofusidie acid phenacyl ester (13-96 g; 20 mmol) in a 1:1 mixture of ethylene glycol mono- and diacetate (80 ml) was added silver carbonate (ll.03 Si 40 mmol).

After being protected from light, the mixture was 1θ stirred for 3 days at room temperature. Insoluble material was filtered off ahd washed with ether (2 x 20 ml). After removal of the solvent from the combined filtrate and'washings at reduced pressure, the liquid residue was diluted with methanol (320 ml), potassium carbonate (5-53 β} 40 mmol) was added, and tlie mixture was stirred for 30 minutes at room temperature. The mixture was evaporated in vacuo. and the oily residue thus obtained was dissolved in a mixture of ether (200 ml) and water (200 ml). After acidification of the stirred mixture with diluted hydrochloric acid, the organic phase was separated and the aqueous phase reextracted with ether (100 ml,). The combined organic extracts wore washed with water until neutral, dried, and evaporated in vacuo. The yellowish amorphous residue thus ob25 tained was purified by dry column chromatography on silicagel (peI.ro 1 etiiii other - ethyl acetate. 6:4) to give 5-54 g “f the desired compound as a colourless amorphous powder which failed to crystallize.

B. 3-0-Acotyl-l6-deacetoxy-l6P-(2'-bromoethoxy)fusidic acid phenacyl ester Phenyl N,Ν-dimcthy 1.formimidate hromi.de (4.6 g; ~ 20 mmol) was added to a- solution of 3-O-acetyl-l6-deacetoxy-l6g-(2’-hydroxyethyloxy)fusidic acid phenacyl ester (4.21 g; 6.2 mmol) in dimethylformamide (25 ml), and the mixture was stirred for lfi^hoUrs at room temperature. After dilution with ether (100 ml), the mixture was washed with water (4 x 25 ml), and the remaining organic phase was dried and evaporated in vacuo. The oily residue thus -obtained was purified by dry column chromatography on silicagel (petroleum ' other-ethyl acetate; 85:15) to give 3.16 g of 3-°“acetyl-l6-deacetoxy-1.6p-(2,-bromoethoxy)fusidic acid phenacyl ester as a colourless amorphous product.

C. 3-0-Acetyl-l6-deacotoxy-l6(3- ( 2 1 -azidoethoxy)fusi.r.lic acid phenacyl ester A solution of 3-0-acetyl-l6-deacetoxy-l6f3-(2'-bromoctlioxy)fusidic acid phenacyl ester ( 1.04 g; 1.4 mmol) and lithium azide (0.34 g; 7 mmoj.) in dimethylformamide (.20 nil) was stirred fur 16 hours at room temperature.

Tiie mixture was diluted with ether (80 mi), washed with water (4 x 20 ml), and the organic phase was dried and evaporated i n vacuo to yield 0.97 g of the desired compound as a foam. IR (KBr) : 2100 cm1·(-N^).

D. l6-Pcacetoxy-ί6p — (2'-azidoethoxy)fusidic acid To a solution of 3-0-acetyl-l6-doacetoxy-l6P-(2'-azidoethoxy)fusidic acid phenacyl ester (0.95 gj 1.34 mmol) in ethanol (20 ml) was added 5 N aqueous sodium hydroxide (2.7 ml), and the mixture was stirred for 18 hours at room'temperature. The solvent was removed in vacuo, and the resulting oily residue was dissolved in water (40 ini) and extracted with ether (20 ml). The aqueous phase was separated, acidified with dilute hydrochloric acid, and the oily precipitate which formed was twice extracted with ether. Tiie combined ethereal extracts wore washed with water, dried, and evaporated to give 0.8 g of an amorphous product which crystallized from diisopropyl ether to yield 0.4.1 g of l6-deacetoxy-16β-(2 *-az.idoethoxy ) fusidic aeid, mp. 179—182°C. Two recrystallizations from the same solvent gave the analytically pure compound, melting point X84-185°C. ‘>5 Example 72 l6-Oeacetoxy-l6P-c!thoxy-24.25-dihydrofusidic acid A. l6-Peacetoxy-l66-ethoxyfusidic acid benzyl ester To a solution of l6-deacetoxy-l6a-bromofusidic acid benzyl ester (3.14 g; β mmol) in ethanol (25 ml) was added silver carbonate (2.76 g; 10 mmol),and, after protection from light, the mixture was stirred at room temperature for 16 hours. The insoluble material was filterod off, washed with ethanol (2 x 5 ml), and the -,(1 combined filtrate and washings wore evaporated in yactio.

The amorphous residue thus obtained was purified by dry column chromatography on silica gel (petroleum ether: ethyl acetate. 6θ:4θ) to yield 1.66 g of the desired compound as a colourless foam.

B. l6-Deacetoxy-l6j3-ethoxy-24,25-dihydrofusidio acid IO/ί Palladium on carbon catalyst (0.4 g) was added to a solution of 16-deacetoxy—Ιδβ-ethoxyfusidic acid benzyl ester (l.2 g;/-\>2 mmol) in ethanol (20 ml), and the mixturo was shaken in a hydrogen atmosphere for 2q 40 minutes. The catalyst was filtered off, washed with ethanol, and tho combined filtrate and washings were evaporated in vacuo. The resulting residue was crystallized from ether to afford 0.92 g of 16-deacetoxy 16P-ethoxy-24,25-dihydrofusidic acid, molting point: . 191-192°C. - 96 43478 Example 7'i 16-Peace Iu\y-1, 21,2 '-trifluoroethoxy)-2^1, 25-dlhydro fus id i.e acid To a solut i on of l6-deacetoxy-l6p-(21,2 * ,2’-tri5 fluoroothoxy)fusidic acid (278 mg; 0.5 mmol) in 96/ή ethanol (5 ml) was added 10j£ palladium on calcium carbonate catalyst (50 mg), and the mixture was shaken in a hydrogen atmosphere for 20 minutes.

The catalyst was filtered off, washed with 96jo ethanol, and the combined filtrate and washings were af evaporated to dryness in vacuo. The residue crystallized from diisopropyl other to afford 220 mg of the desired compound, melting point: 204-205°C. Recrystallization from the same solvent gave the j5 analytical sample, melting point: 204-205°C.

Example 74 l6-Deacot oxy-16β-( 21 -fluoro ethoxy )-24,25-dihydrofusidic acid By substituting l6-doacetoxy~l6p-(2*-fluoroethoxy) 2q fusidic acid for the l6-deacetoxy-l6p-(2’,21,2'-trifluoroethoxy)fusidic acid in the procedure of Example 73 l6-deacetoxy-l6p-(2'-fluoroe thoxy)-24,25-dihydrofusidic acid, melting point: 18O-182°C was obtained. - r7 Example 75 H-Pehydro-l6-deacet;oxy-l6β-ethoxyfusidic acid < To a suspension of 3-0—acetyl—11—dehydro—16—deacetoxy -.l6a~bromof usidie acid phenacyl ester (5-57 £5 8 mmol) in ethanol (60 ml) was added silver carbonate (4.41 g; l6 mmol), and, after being protected from light, the mixture was stirred for 18 hours at room temperature.

Insoluble material was filtered off and washed with ethanol (2 x 20 ml). The combined filtrate and washings containing the crude 3-0-acetyl-ll-dehydro-l6-deacet;oxy -l6p-ethoxyfusidic acid phenacyl ester were diluted with ethanol (80 ml), JS aqueous sodium hydroxide (32 ini) was added, and the mixture was stirred for 20 hours at room temperature. The solvent was removed in vacuo, to the residual oil was added water (lOO mi) and ether (lOO ini), and the stirred mixture was acidified by addition of 4 N hydrochloric acid. The organic phase was separated, the aqueous phase was reexfcracte'd with ether (lOO ml) , and the combined organic extracts were washed with water until neutral, dried, and evaporated in vaeuo. The resuiting oily residue was purified by dry column chromatography on silieagel (ether s petroleum ether : acetic acid, 50:50:0.5), and the yellowish amorphous product thus obtained crystallized from diisopropyl ether to give 2.12 g of 11- dehydro-l6-deacetoxy-l6 g-ethoxyfusidic acid, <)8 4 3 4 7 8 molting point lb6-167°O. Rocrystallization from otherdi isopropyl other afforded the analytical sample, · molting point 167-168°C.

Example 76 ll-Behydro-l6-deacetoxy-l6B- (2r-fluoroethoxy), fusidic acid Following the procedure described in Example 75 but substituting 2-fluoroethanol for the ethanol, 11-dehydro-16-deacetoxy-16 β - ( 2 ’-fluo roethoxy)fusidic acid was obtained as a colourless, amorphous powder.

The compound could be converted into a crystalline sodium salt (see Example 88).

Example 77 3,ll-Bi.sdehydro-l6-deacetoxy-l6B-ethoxyfusidic acid Jones' reagent (12.0 ml) was added dropwise at 15°C to a stirred solution of l6-deacetoxy-l6p-ethoxyfusidic acid (10.24 g of the hemihydrate; 20 mmol) in acetone (200 ml). After the addition was finished, the cooling-bath was removed, and the mixture was stirred for 3θ minutes at room temperature. To the mixture was added ether (300 ml) and water (200 ml), and stirring was continued for 15 minutes. The organic layer was - . separated, the aqueous phase was reextracted with ether (lOO ml), and the combined organic extracts were washed with water until neutral, and dried. On concentration of the ethereal solution to about 100 ml, crystallization of a colourless product began. After being kept in the refrigerator overnight, the crystals were collected, washed with ether, and dried to afford 7.02 g of 3, ll-bi.sd Example 78 3-Behvdro-l6-deacetoxy-l68 -ethoxyfusidic acid A mixture of 3,ll-bisdehydro-l6-dcace-boxy-l6p-et;hoxy fusidic acid (6.05 g; 12.14 mmol), 2-ethyl-2-methyl-l,3 dioxolane' (60 ml), and p-toluenesulphonic acid (0.24 g) was refluxed for 40 minutes on an electric heating-bath. After cooling to room temperature, ether (200 ml) and pyridine (0.5 ml) were added, and the mixture was washed with water (4 χ 5θ ml). The organic phase was dried and evapoi’ated in vacuo to leave 7.06 g -10043478 of the crude 3-ethylene ketal of the 3.11- bisdehydro acid as a gum, which failed to crystallize.

To a stirred solution of the above residue in ethanol (140 ml) was added at 5°C solid sodium borohydride (2 g) in portions. After the addition was finished, the cooling-bath was removed, and the mixture was stirred for 45 minutes at room temperature. The mixture was neutralized with acetic acid, water (420 ml) was added, and the oily precipitate which formed was extracted with ether (2 x 150 ml). The combined organic χ extracts were washed with water (4 x 25 ml), dried, and evaporated in vacuo. The resulting oily residue was crystallized from ether-diisopropyl ether to afford 3·12 g of the 3-ethylene ketal of 3“ dehydro-16-deacetoxy-l6P~ethoxyfusidic acid, melting point 166-169°C. Concentration of the mother liquor furnished another 2.04 g of the desired compound, melting point 166—169 C.

Two recrystallizations from methylene chloride-diisopropyl ether gave the analytical sample, melting point 171-172°C. .

A solution of 3-.dehydro-l6-deacetoxy-l6g-ethoxyfusidic acid 3-ethylene ketal (3.93 gj 7.3 mmol) in methanol (40 ml) was acidified with 2 N hydrochloric -J. 01 acid (2 ml) and refluxed for 20 minutes on the I steam-bath. After cooling, water (l60 ml) was added, and the oily precipitate, which formed, was extracted with ether (2 x 100 ml). The combined organic extracts were washed with water until neutral, dried, and evaporated in vacuo. The resulting amorphous residue was crystallized from ether to yield 2.94 ε of 3-dehydro-l6-deacetoxy-l6{3-ethoxyfusidic acid, melting point 175-177°C. Reciystallization from the same solvent raised the melting point to 177-17^°C.

Example 79 l6-Deacotoxy-l6p-j.sopropylsulphinylfusidic acid β-diethylaminoethyl ester To a solution of the sodium salt of l6-deacetoxyΊ -l6p-isopropylsulphinylfusidic acid (320 mg; 0.5 mmol) in 2 ml of dimethylformamide was added p-(diethylamino)ethyl chloride (0.08 ml; 0.55 mmol). A crystalline product started to precipitate when this mixture was left at room temperature for 5 hours. Water (5 ml) was then added, and the product was filtered off, washed with 5 ml of water and dried to afford 310 mg of l6-deacetoxy-l6p~ isopropylsulphinylfusidic acid @-diethylaminoethyl ester, melting point 156-158°C. -1°2 43473 Example SQ . l6-Deacetoxy-l6g-isopropylsulphinylfusidic acid acetoxymethyl ester To a solution of the sodium salt of l6-deacetoxy5 -l6p-isopropylsulphinylfusidic acid (320 mg; 0.5 mmol) in 2 ml of dimethylformamide was added chloromethyl acetate (0.05 ml; 0.55 mmol). After standing for 24 hours at room temperature, ethanol (5 ml) and water (5 ml) was added to precipitate the desired product as colourless crystals, which were filtered off, washed with water (5 ml), and dried to give 290 mg; melting point 151-153°C. Recrystallization from ethyl acetatepetroleum ether raised the melting point to 152-154°C.

Example 81 l6-Deacetoxy-l68-isopropylthiofusidic acid acetoxymethyl ester By following the procedure described in Example 80 but substituting l6-deacetoxy-l63-isopropylthiofusidic acid for l6-deacetoxy-l6p- isopropylsulphinyl20 fusidie acid l6-deacetoxy-l63-isopropylthiofusidic acid acetoxymethyl ester was prepared as colourless crystals of melting point 77-83°C, - 1 03-i ........) 2 4 7 S Examples 82-89 « Sodium salts of Ιόβ-ethers, l6B-thioethers and 16(3-alkylsulohinyl compounds of l6-deacetoxyfusidic acid and its 3- and 11-dehydro derivatives.

Crystalline sodium salts of the compounds described in Examples 1, 35, 46, 48, 50, 75» 76, and 78 were obtained by the following procedure: A solution of the corresponding acid (lO mmol) in methanol (25 ml) was titrated with 2 N methanolic sodium hydroxide using phenolphthalein as an indicator. After evaporation to χ dryness in vacuo, the oily or amorphous residue thus obtained was taken up in acetone (ca. 100 ml), the resulting solution was concentrated to about half the volume, and upon scratching the desired sodium salt began to crystallize. The mixture was kept for 2 hours at room temperature, thereafter the crystals were collected, washed with acetone, and dried to give the pure sodium salt of the desired compound.

The sodium salts prepared by this method are listed in table XVII. Microanalysis, IR- and NMR data obtained for these compounds are in agreement with their structure. ί_104_ i 3 4 7 S Examplefil «2 Aβ3 Acid, described in Example 82 H,a-0H H,a-0H s ch(ch3)2 1 83 H,a-0H H,a-0H S-> 0 CHtcH3)2 35 84 H,a-0H H,a-0H 0 ch2ch3 46 85 · H,a-OH H,a-0H 0CH2CF3 48 86 H,a-0H H,a-0H 0CH2CH2F 50 87 H,a-OH 0 0 ch2ch3 75 88 H,a-0H 0 0CW 76 89 0 H,a-0H 0 ch2ch3 78 Example 90 Potassium salt of 16-deacetoxy-l6g-(21-hydroxyethoxy)fusidic acid A solution of l6-deacetoxy-l6(3-(2*-hydroxyethoxy)15 fusidlc acid (2.64 g; 5 minol, calculated as the hemihydrate) in methanol (iO ml) was titrated against -10543473 phenolphthalein with 2 N methanolic potassium hydroxide. After evaporation to dryness in vacuo, the amorphous residue thus obtained was dissolved in methanol (2.5 ml), acetone (60 ml) was added, and the mixture was con— centrated to about 15 ml of reduced pressure. Colourless crystals precipitated on scratching,· were filtered off, washed with acetone,'and dried to afford 2.32 g of the desired compound.

Example 91 . ' l6-Deacetoxy-3-d.ehydro-l68»isopropylthio-24i25-dihydrofusldlc acid By following the procedure of Example jh but substituting l6-deacetoxy-l6p-isopropylthio-24,25dihydrofusidic acid acetoxymethyl ester for 1615 deacetoxy-l6P-isopropylthiofusidic acid acetoxymethyl ester, l6-deacetoxy-3- dehydro-16 β-isopropylthio24,25-dihydrofusidic acid was prepared.

Example 92 ll-Dehydro-l6-deacetoxy-l6p(2t,21,2*-trifluoroethoxy) fusidic aoid Following the procedure described in Example 75 but substituting 2,2,2-trifluoroethanol for the ethanol, 11-dehydro-l6-deacetoxy-16 β—(2 *,2*,2*-trifluoroethoxy)fusidic • acid was obtained. i » ' '-106-.* ·. j 2 4 7 8 Example 93 Cream 16-Deacetoxy-Ιόβ-isopropylthio-24,25dihydrofusidic acid................. 20 g Petrolatum.......................... 150 g Liquid paraffin................... 150 S Spermaceti ........................ 50 g Sorbitan monopalmitate ............ 50 g Polyoxyethylene sorbitan mono10 palmitate ......................... 50 g Water ............................. 530 g 1000 g Heat petrolatum, paraffin, spermaceti, sorbitanmonopalmitate, and polyoxyethylene sorbitan monopalmitate to 70°C and slowly add the water at 72°C with agitation. Continue agitation until the cream has cooled. Triturate l6-deacetoxy-l6β-isopropylthio24, 25-dihydrofusidic acid into the cream base and homogenize using a roller mill. Fill the cream into laquered aluminium collapsible tubes.

Example 94 Ointment l6-Dθacetoxy-l6β-isopropylthiofusidic acid sodium salt ..................... 20 g Liquid paraffin ...................... 13θ g Cetanol .............................. 4 g Lanolin anhydrous .................... 46 g Petrolatum........................... 792 g 1000 g -1071 i Melt paraffin, cetanol, lanolin, and petrolatum at 70°C. After cooling to below 4o°C, triturate 16deacetoxy-l6p-isopropylthiofusidic acid sodium salt. Fill the ointment into laquered collapsible aluminium tubes. , Example 95 Ointment l6-Deacetoxy-l63-isopropylsulphinyl fusidic acid sodium salt ........... 10 g Liquid paraffin .................... 1β8 g Cetanol ............................. 4 g Lanolin anhydrous ................. 46 g Petrolatum......................... 802 g 1000 g Melt paraffin, cetanol, lanolin, and petrolatum at 70°C. After cooling to below 40°C, triturate l6deacetoxy-l63-isopropylsulphinyl fusidic acid sodium salt. Fill the ointment into laquered collapsible tubes.

Example 96 Capsule ll-Dehydro-l6-deace,fcoxy-l6i3-e'bhoxyfusidie acid sodium salt ...................... 250 g Microcrystalline cellulose ............ 145 g Magnesium stearate .................... 5 S 400 g I 1084S4 7S Pass the ingredients through a 60 mesh sieve and mix for 10 minutes. Kill the mixture into hard gelatin capsules No. 00 (Parke Davis & Co.) using a capsule fil weight of 400 mg.

Example 97 Preparation of tablets l6-Doacetoxy-l6p-(2 *,2 *,2’-trifluoroethoxy) fusidie acid................. 250 g AvicolpH 101 ....................... 120 g STA-Rx 1500 .................Z....... 120 g Magnesium stearate.................. 10 g •iHiAvicel is a registered Trade Mark. ’’ l6-Deacetoxy-l6p-(2',2',2’-trifluoroethoxy)fusidic acid, Avieel and STA-Rx are mixed together, sieved through a 0.7 mm sieve and thereafter mixed with the magnesium stearate. The mixturo is pressed into tablets each of 500 mg.

Example 96 Preparation of suspension 3-Dehydro-l6-deacetoxy-l6grisopropylthiofusidie acid......................... 5· 00 g Citric acid.......................... 0.45 g -109- ι 478 Sodium monohydrogenphosphate ...... 0.70 g Sucrose ........................... 25.00 g ιι I· Tween 80 .......................... 0.05 g Potassium sorbate ................. 0.20 g Carboxymethylcellulose-Na ......... 0.50 g Purified water.................... qs to 100 ml suspension ••Tween” is a registered Trade Mark.

The crystals are micronized and suspended in a solution of the citric acid, the sodium mortchydrogen— phosphate, the sucrose, the potassium sorbate and the Tween80 in 5θ ml water, if necessary under slight warming The carhoxymethylcellulose-Na is dissolved in 20 ml of boiling water. After cooling, it is added to the other ingredients. The suspension is homogenized in a blender and finally purified water is added, to a total volume of 100 ml.

Example 99 Ointment Ai l6-peacetoxy-l6p-isopropylthiofusidic acid sodium salt ........... 20 g B: One of the steroids: hydrocortisone,triamcinolone or fluocinolone....... 10 g Liquid paraffin ........ 138 g Cetanol ............................ 4 g Lanolin anhydrous .......... 46 g Petrolatum..................'....... 802 g 1000 g ;-11042478 Melt paraffin, cetanol, lanolin, and petrolatum at 7O°C. After cooling to below 4o°C, triturate A and B. Fill the ointment into laquered collapsible tubes.

Example 100 Ai l6-Deacetoxy-l6p-(2 -fluoroethoxy) fusidic acid ........................ 125 S B; One of the antibiotics: Amoxycillin, Cephalexin, Rifamycin, Rifampicin, Clindamycin or Lincomycin, Erythromycin, Pivmecillinam....... 125 S Microcrystalline cellulose ...... 145 g Magnesium stearate ............... 5 S 400 g Pass tho ingredients through a 60 mesh sieve and mix for 10 minutes. Kill the mixture into hard gelatin capsules No. 00 (Parke Davis & Co.) using a capsule fil weight of 4-00 mg.

« Example 101 Ointment A: Tetracycline ................. 15 g B: l6-Deacetoxy-l63-ethoxyfusidic acid ............... 15 g Liquid paraffin ................. I38 g -11143478 Cetanol ......................... 4 g Lanolin anhydrous......... 46 g Petrolatum ............... 782 g Melt paraffin, cetanol, lanolin, and petrolatum at 5 70°C. After cooling to below 4o°C, triturate A and B.

Pill the ointment into laquered collapsible aluminium tubes.

Claims (15)

1. A compound of the following formula I:

in which the C„. _ bond is a single or a double bond, jj

5 and in which and each stand for jjq'Z' 1 ' or oxygen ; A stands for oxygen, sulphur or a sulphinyl radical; stands for a straight or branched alkyl radical which has from 1 to 8 carbon atoms, and which is unsubstituted or substituted with one or more halogen atoms or hydroxy, alkyloxy, aralkyloxy, aryloxy,

10 alkanoyloxy, aralkanoyloxy, aroyloxy, sulfhydryl, alkylthio, aralkylthio,araylthio, alkanoylthio, aroylthio, azido, nitro, cyano, thiocyanato hydroxycarbonyl, alkyloxycarbonyl, aryloxycarbonyl, amino, alkylamino, dialkylamino, arylamino, alkanoylamino, and aroylamino groups; an alkenyl or alkynyl

15 radical having from -2-3to 6 carbon atoms; a cycloalkyl radical having from 3 to 7 carbon atoms in the alicyclic ring, or a mono- or dihalo, lower alkyl, lower alkoxy or hydroxy

- 113 43478 substituted analogue thereof; an aryl, aralkyl or heterocyclylalkyl radical, which is unsubstituted or substituted with one or more halogen atoms or nitro, lower alkyl, hydroxy or alkoxy groups; or an unsubstituted or methyl-substituted

5 heterocyclic radical having 5 or 6 ring atoms and containing oxygen, sulphur or nitrogen atoms; and pharmaceutically acceptable salts and easily hydrolyzable esters (as hereinbefore defined) thereof.

2.

A compound of formula I, in which the C

24-25 bond is ) a single or double bond, and each stand for

H,

H0A stands for oxygen or sulphur and has the meaning defined in claim 1.

- 114 43478

3. A compound of formula I, in which the Cg^ g^ bond is a single or a double bond, Q^ and Qg both stand for *, A stands for oxygen; stands for a straight or branched alkyl radical having from 1

5 to 4 carbon atoms and being optionally substituted with halogen atoms, hydroxy or azido groups, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof,

4. · A compound of formula .1, in which tho Cg^ jo bond is a single or a double bond, Q^ and Qg both stand for 5 A stands for sulphur; stands for a straight or branched alkyl radical having from 1 to 4 carbon atoms and being optionally substituted with halogen atoms, hydroxy or azido groups, and pharma15 oeutically acceptable salts and easily hydrolyzable osters theroof.

5. · A compound of formula I, in which the Cg^ g_ bond is a single or a double bond, Q^ and Qg both stand for 5 A stands for a sulphinyl radical;

HU'

20 R^ stands for a straight or branched alkyl radical having from 1 to 4 carbon atoms and being optionally substituted with halogen atoms, hydroxy or azido groups, and pharmaceutically acceptable salts and easily hydrolyzable osters theroof.

115

6. . A compound of formula I’, in which the bond is a single or a double bond, one of and Q„ is oxygen and the other is , A stands for oxygen; stands for a straight or branched.

5 alkyl radical having from 1 to 4 carbon atoms and being optionally substituted with halogen atoms, hydroxy or azido groups, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof,

7. A compound of formula I, in which the 2 ^

10 bond is a single or a double bond, one of and

Q 2 is oxygen and the other is HO'''* ’ A s ^ ands for sulphur; R^ stands for a straight or branched alkyl radical having from 1 to 4 carbon atoms and being optionally substituted with, halogen atoms, hydroxy

25 or azido groups, and pharmaceutically acceptable salts . and easily hydrolyzable esters thereof,

8. , A compound of formula Ϊ, in which the 2 ^ bond is a single or a double bond, one of and Q 2 is oxygen and the other is iio·' 1 '’ s ^ and s for

20 sulfinyl; R^ stands for a straight or branched alkyl radical having from 1 to 4 carhon atoms and being optionally substituted with halogen atoms, hydroxy or azido groups, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

- 116

9. l6-Ileacetoxy-l6p-isopropyltliiofusidic acid and its C 2 ; ( _2 3 -dihydro analogue and pharmaceutically acceptable salts and easily hydrolyzable esters thereof'.

1(1, J,6-I.>oacotoxy-l6p-isopropy Jsulphinyll’usldic acid 5 and its Ερή ihydro analogue, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

11. 1 .l~Dehydro-l6-deacetoxy-16B-isopropylthiofusidic acid, and its ^-diliydro analogue, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

10. 12. 3- Dehydro-l6-deacetoxy-l6β-isopropylthiofusidic acid, and its C o ^ ^-dihydro analogue and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

11. 13· l6-Deacetoxy-l6p-ethoxyfusidic acid and its 11—dehydro analogue, and pharmaceutically acceptable salts and easily

Ij hydrolyzable esters thereof.

12. 14· 16-Deacotoxy~l6p~(2'-fluoroethoxy)fusidic acid and its 1.1.- dehydro analogue, and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

- 117 43478

13. 15- l6-I)oacetoxy-ΐδβ-(2,2,2 -trifluoroethoxy)fusidic acid and its 11-dehydro analogue; and pharmaceutically acceptable salts and easily hydrolyzable esters thereof

14. 16. ΐ6-Ββηοβΐοχν-ΐ6β-(ΐ,3 -difluoroisopropyloxy)

5 fusidic acid and pharmaceutically acceptable salts and easily hydrolyzable esters thereof.

15. 17. A method for the preparation of a compound of formula I oxygen; A stands for oxygen, sulphur or a sulphinyl radical; R^ stands for a straight or branched alkyl radical which has from 1 to 8 carbon atoms,

- 118 and which is unsubstituted or substituted with one or more halogen atoms or hydroxy, alkyloxy, aralkyloxy, aryloxy, alkanoyloxy, aralkanoyloxy, aroyloxy, sulfhydryl, alkylthio, aralkylthio, arylthio, alkanoylthio, aroylthio, azido, nitro, cyano, thiocyanato, hydroxycarbonyl, alkyloxycarbonyl, aryloxycarbonyl, amino, alkylamino, dialkylamino, arylamino, alkanoylamino, and aroylamino groups; an alkenyl or alkynyl radical having from-23to 6 carbon atoms; a cyeloalkyl radical having from 3 to 7 carbon atoms in the alicyclic ring, or a mono- or dihalo, lower alkyl, lower alkoxy or hydroxy substituted analogue thereof; an aryl, aralkyl or heterocyclylalkyl radical, which is unsubstituted or substituted with one or more halogen atoms or nitro, lower alkyl, hydroxy or alkoxy groups; or an unsubstituted or methyl-substituted heterocyclic radical having 5 or 6 ring atoms and containing oxygen, sulphur or nitrogen atoms: and pharmaceutically acceptable salts and easily hydrolyzable esters (as hereinbefore defined) thereof, comprising reacting a compound of formula Vb aralkanoyl or an aroyl ; is as defined above, Y stands for chlorine, bromine or iodine; and stands for a straight or branched alkyl radical haying from 1 to 6 carbon atoms, an unsubstituted or substituted aralkyl radical an alkanoylmethyl or aroylmethyl radical, an alkanoyloxyalkyl or aroyloxyalkyl radical,

- 119 4S473 an alkyloxymethyl or a cyanomethyl radical; with a compound of formula VH: Rj-A-H, in which R x is as defined above and A represents oxygen or sulphur, yielding a compound of formula VUI:

in which .Q^, Q 2 , R^ and R^ are as defined above, and A is oxygen or sulphur, which compound is thereafter subjected to a hydrolysis, if desired,or necessary and a com pound of formula X, A being sulphur, optionally is

IO oxidized to a compound of formula I, A being a sulphinyl radical; and the free acid can be transformed into a pharmaceutically acceptable salt or an easily hydrolyzable ester thereof.

16. 18. A method according to claim 1 7, in which a

15 compound of formula I, wherein and/or Qg aro

TT , is oxidized to a compound of formula I wherein •and/or Qg are oxygen.

- 120

17. 19. A method according to claim 17 in which a compound of formula X, having a double bond between C-24 and C-25, is hydrogenated to a compound of formula I having a single bond between C-24 und C-25.

5 20 A method for the preparation of a compound of formula ·Ι as shown in claim 17 in which R stands for an aryl radical and A is sulphur, comprising reacting a compound of formula XV, ι .

10 in which Q x , Q g and R^ are as defined m claim 17, and the OH group at C-l6 is α-oriented, with a phosphine and a compound of formula R^SSR^, thereby obtaining a compound of formula VIII shown in claim 17, in which A is sulphur and is aryl and () 1 1, Q 2 and R^ are as defined in claim 17, which

15 compound is thereafter transformed into a compound of formula I as defined in claim 17.

121

4347 8

21. A method for tho preparation of compounds of formula I as shown in claim 17, comprising reacting a compound. of formula IV as shown in claim 20,in which the hydroxyl , 1 group at C-16 is α-oriented and Q 1 is different from

J£

5 the group i with a reactive derivative of an alkylsulphonic or arylsulphonic acid, to form a compound of the general formula IX:

in which Q^j R^ and the dotted line between C-24

10 and C-25 are as defined in claim 17, Q’

H stands for oxygen or the group Rg being an alkanoyl, aralkanoyl or aroyl radical, and represents an alkylsulphonyl or arylsulphonyl radical, which compound is reacted with a compound

15 R^-A-H, R^ being as defined in claim 17 and A being oxygen or sulphur to form a compound of formula VIII, which compound is thereafter transformed into a compound of formula I as defined in claim 17.

122

22. A modification of the method according to claim 17 where a compound of formula VITI in which Q’ and Q„ each stand for 1 z the group or oxygon, and represents an-unsubstituted or substituted benzyl radical, a cyano5 methyl, alkanoylmethyl or aroylmethyl radical is converted into a compound of formula I by reduction.

23. A method according to claim 17 comprising converting a compound of formula VIII in which Q^, Qg, R^, and the dotted line between C-24

10 and C-25 are as defined in claim 17, A is oxygen, sulphur or a sulphinyl radical, and R^ stands for a hydroxy-substituted alkyl radical, into a corresponding compound in which R^ stands for a halo-substituted alkyl radical by treatment

15 with a halogenating agent, thereafter reacting the halo-substituted alkyl derivative of formula VIII with an aliphatic or aromatic alcohol,with an aliphatic or aromatic mercaptan, with ammonia or an aliphatic or aromatic amine, or with a salt of a

18. 20 .lower alkanoic acid or benzoic acid,with silver or sodium fluoride, an alkali metal azide, nitrite, cyanido or thiocyanate or with a salt of a lower thioalkanoic acid or thiobenzoic acid forming a compound of formula VIII :i.n which Q ± , Q 2 , R^, A,

123 43478 and tho dotted line between C-2h and C-25 have the meaning as defined above, and stands for an alkyl radical substituted by a fluorine atom, an alkyloxy, aralkyloxy, aryloxy, alkylthio, aralkylthio, arylthio,

5 amino, alkylamino, dialkylamino, azido, nitro, cyano, thiocyano, alkanoyloxy, aralkanoyloxy, aroyloxy, alkanoylthio or aroylthio radical, which compound is thereafter transformed into a compound of formula I as defined in claim 17.

10 24. A pharmaceutical composition containing as the active ingredient a compound of claim 1 and comprising a solid or liquid pharmaceutically carrier' or diluent.

25« A pharmaceutical composition in dosage unit form for systemic treatment of bacterial infections in patients

15 which comprises as a therapeutical ingredient at least one compound of claim 1, the therapeutically active compound being admixed with a non-toxic, pharmaceutically acceptable carrier, and the dosage unit being between 100 to 1000 mg, preferably 100 to 500 mg,

2o calculated as the free acid of the therapeutically active compound.

26. A pharmaceutical composition as claimed in claim 25 wherein the dosage unit is from 200 to 750 mg calculated as the free acid of the therapeutically active compound.

124 43478

27· A pharmaceutical composition in dosage unit form for topical treatment of bacterial infections in patients which comprises as a therapeutical ingredient at least one compound of claim 1, the therapeutically

5 active compound being admixed with a non-toxic, pharmaceutically acceptable carrier, and the dosage unit boing from 0.1 mg to 10 mg per sq. centimetre of the infected area.

28. A pharmaceutical composition as claimed in claim 25 or

10 26 wherein the dosage unit is in tho form of tablets.

29. A pharmaceutical composition as claimed in claim 25 or 26 wherein the dosage unit is in the form of capsules.

30. A pharmaceutical composition as claimed in claim 27 wherein the dosage unit is xn the form of a cream.

15 31. A pharmaceutical composition as claimed in claim

27 wherein the dosage unit is in the form of an ointment.

32. A pharmaceutical composition as claimed in claim 25 or 26 wherein the dosage unit is in the injectable form of preparation.

125

43 47 8

33 · A pharmaceutical composition as claimed in claim 25 or 26 wherein the dosage unit is a suspension for oral use and containing the therapeutically active compound in ar amount of· from 2 to 25 per cent.

5 34· A pharmaceutical composition according to claim

19. 24 in which the active ingredient is one of the compounds claimed in claims 9—10.

35 · Λ pharmaceutical composition according to claim 27, which comprises as the therapeutic ingredient a

10 mixture of a compound of claim 1 and a corticosteroid.

36. A pharmaceutical composition according to claim 27> which comprises as the therapeutic ingredient a mixture of a compound of claim 1 and tetracycline

37 A pharmaceutical composition according to claim 25 to 25 26, which comprises as the therapeutic ingredient a mixture of a compound of claim 1 and an antibiotic .selected from the group consisting of penicillins, cephalosporins, rifamycin, erythromycin, lincomycin and clindamycin.

126 43478

38. A method of treating bacterial infections in nonhuman animals which comprises administering into the body a member of the compounds of claim 1 in an amount of from

0.25 g to 4 g per day.

5 39· A method according to claim 38 in which the therapeutically active compound is administered by the oral route, in amounts from 0.5 to 3 g per day.

40, A compound of the formula defined in Claim 1 substantially as hereinbefore described in any one of Examples 1 to 92 of

10 the foregoing Examples.

41. A method for the preparation of a compound of the formula defined in Claim 1 substantially as hereinbefore described in any one of Examples 1 to 92 of the foregoing Examples.

15 42. a pharmaceutical composition substantially as hereinbefore described in any one of Examples 93 to 101 of the foregoing Examples.