DK143905B - R ANALOGY PROCEDURE FOR THE PREPARATION OF FUSIC ACID DERIVATIVES - Google Patents

Description

(19) DENMARK

® (12) PUBLICATION OUT 11) 3905 B

DIRECTORATE OF THE PATENT AND TRADEMARKET SYSTEM

(21) Application No. 2857/76 (51) (nt.CI.3 C 07 J 13/00 (22) Filing date 25 · Jun. 1976 C 07 J 31/00 (24) Running day 25-Jun. 1976 ® J * 1/00 (41) Available Available Dec 26, 1976 J A3 00 (44) Submitted Oct. 26, 1981 (86) International Application No. - (86) International Filing Day (85) Continuation Day - (62) Stock Application No. -

(30) Priority 25 · Jun. 1975, 26989/75 * GB Nov 7 1975 * 46229/75, GB

(71) Applicant LEO PHARMACEUTICAL PRODUCTS LTD. A / S (INVENS CHEMICAL FACTORY PRO = DUCTION SHARE COMPANY), Industrial Park 55 * 2750 Ballerup, DK.

(72) Inventor Welf von Daehne, DK: Poul Rødbroe Rasmussen, DK.

(74) Plenipotentiary - (54) Analogous procedures for the preparation of fusidic acid derivatives.

The present invention relates to an analogous process for the preparation of novel derivatives of fusidic acid and pharmaceutically acceptable salts thereof, which is characterized by the characterizing part of the claim. The new compounds have the general formula I

H + C CH

new

D zk 0 f 6

J [. COOR

, 4rtJ

1 AV

: h C «, 2 143905

. in which the bond between C-24 and C-25 is single or double, 12H

Q and Q represent a group or oxygen, A represents oxygen, sulfur or a sulfinyl group, R "*" represents a straight or branched alkyl group having 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, which alkyl groups may be mono-, di- or tri-substituted with halogen atoms or with hydroxy, alkoxy, aralkoxy, aryloxy, alkanoyloxy -, aralkanoyloxy, aroyloxy, sulfhydryl, alkylthio, aralkylthio, arylthio, alkanoylthio, aroylthio, azido, nitro, cyano, thiocyano, hydroxycarbonyl, alkyloxycarbonyl, aryloxycarbonyl -, alkylamino, dialkylamino, arylamino, alkanoylamino or aroyl amino groups,. wherein the alkyl-containing substituents may contain from 1 to 4 carbon atoms in the alkyl group, and wherein R 1 carbon atoms in the alicyclic ring, such as a cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl group, which cycloalkyl group may be optionally mono-, di- or trisubstituted by halogen atoms, C -C 2 -alkoxy or hydroxy groups, and wherein R 2 may further mean a phenyl or phenyl - C -C ^ alkyl, hydroxy or alkoxy groups having 1 to 4 carbon atoms, a heterocyclic or a heterocyclyl-C C-C ^ alkyl group, wherein the heterocyclic moiety contains 5 or 6 ring atoms, of which up to 4 can be nitrogen and / or 1 may be oxygen or sulfur, which heterocyclic rings may optionally be substituent with a methyl group such as a 2- or 3-pyrrolyl, 2- or 3-pyridyl, 2- or 3-thienyl, 2-, 3- or 4-pyridyl, 2-, 4- or 5- pyrimidyl, 2- or 3-pyrazolyl group, an imidazolyl group, eg a 1-methyl-2-imidazolyl group, a triazolyl group, eg a 5-methyl-1,2,4-triazol-3-yl group, a tetrazolyl group, e.g. a 1-methyl-1H-tetrazol-5-yl group, a thiazolyl group, or a thia-diazolyl group, e.g., a 5-methyl-1,3,4-thiadiazol-2-yl group, R 2 represents hydrogen or a C -alkanoyloxy-C ^-Cy alkyl, phenyl - C ^-Cg-alkanoyloxy-C ^-C ^-alkyl, benzoyloxy-C ^-C ^-alkyl, C ^-Cg-alkoxycarbonyloxy-C ^ -C ^-alkyl, C ^-C ^-lactonyl or di-C ^-Cg-alkylamino-C,-C „-alkyl group, 1-> 12 Of particular interest are compounds in which Q and Q are both

H

represents or one of them represents oxygen, A is as defined above, and R represents a straight or branched alkyl group 3 143905 having from 1 to 4 carbon atoms which may be optionally substituted by halogen atoms, hydroxy groups or an azido group and the bond C-24 - C-25 is single or double.

Of particular interest between these groups are those wherein R 1 is ethyl or Isopropyl optionally substituted with fluorine.

As pharmaceutically acceptable, non-toxic salts, there may be mentioned alkali metal salts and alkaline earth metal salts, e.g., sodium, potassium, magnesium or calcium salts, as well as ammonium salts or suitable non-toxic amine salts, e.g. 2-hydroxyethylamine, bis- (2-hydroxyethyl) -amine or tri- (2-hydroxyethyl) -amine, cycloalkylamines, eg dicyclohexylamine, or benzylamines, eg N, N * -dibenzylethylenediamine, and -dibenzylamine. For some purposes, silver salts can also be used, especially for local treatment.

Easily hydrochloric acid, and or lactonyl esters such as phthalidyl esters, or dialkylaminoalkyl esters such as diethylaminoethyl esters.

It is well known that the fusidic acid itself possesses antibacterial properties and it is also known that variations in structure can cause a total loss of these properties. However, it has now been found that the compounds of the present invention exhibit both in vitro and in vivo interesting antimicrobial and pharmacokinetic properties, and that the compounds can therefore be used in the treatment of bacterial infections in humans and animals. For example, in vitro studies have shown that the substances are highly effective against a variety of bacteria, e.g. , Λ o • d -Η (0 Ο -Η Μ h η <! Ej σ '\

© · Η Η OOOV3CMOOO ΕΛ Ο Ό VD

-r ^ {^ 3 ····· «« · ·· · ·

HacJO 1Λ Ρ} - Η Η ΕΛ CM ΙΛ Η Ο Ο C \! CM Ο Η Ε d h Ο Η Ο η <η j 60 ---——-—-—-----——---- 3- (0 Μ ^ d · Η <Λ

Η Η <* 00 CM

5Ρ η η μ woovowmiammmnw-a · ο

g · Η Ρ ΟΟΡΓΗΟΟΟΟΟΟΟΟΟ CM

Η Ο, Ώ Ο .............

pendant οοοοοοοοοοοοοο ο 1 a α - * ____ -Λ ® Ρι Ο CM Ο C \ ~ © oHOinvoc ^ oocvimoow co ho «..............

° ρ © Ο Hooooo-H «ncncMj-Hn η ^ CO ^ Η ___ rt ι Η ώ Ο .οι> ° Η Β Ém * O ^ hO ^ OOWO Ο OOC \ h n2 ft O CUOWHCVlO-dO-OVOVOiOiO i> o 2 Island Island ............

tH S h £ OOOOOOOHHOHOO O

t * P O O

<H '° CQ fcOH

d fc -.-.- ^ (2 j O Ή d o ® Γ. <d O fp

H m · t-Ι CO

® 2 a i> wvoocoooiAOomvoin cv d «ftiiri nHin ^ -cMj- ^ j-NoiotWHfvi ch d .2 d d οοοοοοοοοοοο o h R p © o .............

-g CO u o οο οοοοοοοοοοο o S h -: ----

'Η'πίΛω P P P P

φ CM S HPHPHPH1I I II I

O 0 - H © Η © Η O H O

® £ ¢ £ 0.00,0 = 2 = = = = «o cm d £ w £, y £, ¾ £ oW1 ^ o ^ odoflorriiii“ O i! tJ O tj O Ό o - T3 'd w g CM CM CM CM CM CM CM ^

Tj ^^ (=, 1¾ (=,

f, cncnc ^ co <nr \ coc ^ c ^ icMCMcocM

3 aaaffiHKaKWHKfca ® - Ά οοοοοοοοοοοοο, -— '' - '-' - - - CM CM CM CM CM w

uh ffiWKKKKOKffiKKKS

H o οοοοοοοοοοοο ο ο u £ ------

^ 0 <J C0CQ °° CQMC0COOOOO

od tn m cd ----

£ -H

'T 4J> 7 * »T *» τ' · »T *« - H- Hh l-H HH

• ri * J-M HH HH H- »—- HH H-I HH

-P B .. oooo O O O o o

d £ fill II I II

<l do1 aaaaooaa oaoaa CO «<, ·» «* n ·> 'T * Vr' * T * HH - tr * * —I HH H“ t HH HH> £ h - t— * Hh hi hn HH HH M HH HH Η »HH« -Η I— «M HH HH HH * - · HH HH HH I * 1- H> HH HH HH V— <> —t. oooooc oooooo

CV I 1 I I I I I I I I I I

^ aaaaaaoaaaaaa awKsæaT saasaa H ~

ISLAND

ft ·· I '· lo HH ^ OO HCMJ-cnoO-iVDO "J i £ _j_______lu: 5 143905

W

o \ O O

hw n r ^ - cv m m * n n cm cm vo cm o vo o Jf)

S o fo HOO ^ OWH CM CM H CM -3 · P £ M

\ u (=, ooooooo o o oooooom time # # ···· * * ···· **

i X O OOOOOOO O O OOOOOOO

w. O v O vJ________ kO --- 'a i • H Cd C \!

c! U

S <η <! o Ό ° O W

^ • 05 ^ 3 ** ·· ·

Xj P -ri Ό CM Ο O CM MO ^^ O

OHO H VOHHVOHOHH Η1Λ

tn Cd Η Θ H

- m &> - 3______> P. T "o in cm

«Π · Η 0 \ O

• P ^ CMr ^ OiOOOOCM CM O CM o _

tH ^ 5M SO O O 1Λ CM CM -3 "MO vo VOIMVO-cJ-OO

ffl tn O O o O O O O O C O Η O O O H O

O · vi 0) (0 FQ i— {i-3 H ~ - '1: u-j O · ° »d P <ON ^ h p | ®cMONOr ^' 'over'0i: vi ° w oo \

jj O £ S g ^ «O H c * o« ή, M OOOO ^^ O

O U '"

Ch O 0} C-j C] r. h n O O VO ^

a 5Hft§HO ^ t0iV0iMHOV0 m \ D O CM V £) CM CM

* 1 * K l «a ·· **** · · · · · * · ^

in fi ^ oo CM Ο O '“i ° ° w 1-1 w rH H CM O CM O

c O P >> Φ tn · η cn aj H p --------------- φ (ti pq ^^ nO _ • h e ^ -cvjincviOvoOOO cm o cm cm o \

ja -g Hc ^ oolr'ONfv | -;: i · CM H- Ma vo t> O O

o β ^ ^ o o o o O O o o o o o o o o «-r O

^ O ft ffl «, W cd rH o

'H p; d O

05 CO CtJ i — l ____________________________ p ~~~ 33 p i3 +5 i3 +3 + 3- P P P P p T3 ^

φ 'f} 2? hhpHHHHH HHHHHHHH

P m m p 0 0 0 0 o Φ ΦΟΦΦΟΟΦ • ri J'pj πηφ, α, α ^^, α, ο, α, ο, α, α, α-α ^

> O ', g iJ.QflrQrQ / l'OP

• H 9 3 OOdOOOOO o ooooooo +3 I, -ri Tj-r (®T5T3'ardT3 • dfl'a'd'd ^^ 1 M - £ 2-5 --------: - --- cd

H

• h m ^ p s ® ® W CM ^ CM CM ^ O ^ • p ch ch ch ^ J5 M ^ X X O ~ CH / i M Kffiffi cm cm γ o / ¾¾. o 1Λ α ω 9 a /

ni 't oooffiffiwoolf — v cm ffi ti cm g o_z I

Æ ίϊ U W CM O O W OJ / \ K vo «ffi \ 1 • H <0 w ϊ ^ M S S f O O O O, r> _w \

P P I CM CM CM CM W g / CCCJZ

fl S U O O CM CM CM KKffiKyjO / "^ <0 x ~ x o o o o o ^ / S o o o g p_____________o__ p <; MWWOTcncnw ω ω cncocflcncocow tn -----—-——-- • Q ajKK h = ffirdBK'KKW-æ

3 o o O OOOO OO OO OOOO

W CM III III I I I I I 'I * Λ Λ o' aaao «« 3 «a o a« a d a «_ ___hd it! ίτΐ_nq Ξ S_S_E__2S_E-E-3- £ -3 — g--

µ µ µ µ s in s trlKKEE ^ E

h ooo oooo ooooooo? O Iliolll 1 1 lllll'l aaa aao a a a a a a a ° a xxx E s s x kxw'ke ke w.incq cm æ s n pqnoca as 7] CM CM CM O 00 CM CM ΙΛ H w

_____jxj i? C ___ H

6 143SOS

(η ^ '· Η Η W ρ * \ Ε p ft oovoooocncnNovooc ^ oo m

> OHHfe-fefeHOOiAHO O

5? O oooooooooooo o I X o ............

'· ® oooooooooooo o O i-l _____ so

fl I

• H Hi W

C ^ .inow o ό Ξ Ή · · · * ·

Sj * tch) WOVOOC'iOOC'SOOOCV] h

Λ ft -fe o CM CM O Η H O CM

2 H 0 Η Η H

tn cd -H o). Λ - m sos______1_il_____ 0 i c <n 1Λ -ri fe <ί 1Γ \ ΡΛ cn ft £ «ΟΜΝΟΟΡΛΙΛΝΟΟΟ ^ O NO o

W 3 O O »Λ NO CM fe fe- NO NO CM o CM

tn tn o .............

® fe ft ® OOOO OOOOOfeOO O

in Η H S___ __

H

H 0

° ld ft o C \ i ΡΛ VD

fe £ 2 © CM. . ..

2 fj? "Fe-NOOCMOOPNNONOOONO H

s ° -P Οΰ fe CM C-N fe fe H fe Η H

k m fe fe 0 k -------- "-'-- <h o tn fe Φ

ft, · S O ON

(S H ft O fe NOOOOOrNcomCMOCMNO i> ffl 0) SO ............

3 S ^ o o feHm ^ fcvifeowcnfe-cnfe o O O fe>. (!) tn -η ω ft fe fe fe ----- CD 5i fe E k oo

S fe! N

• fe H o ΡΛ CM CM

fe ® »TivocvioococnmootnrNcn cn h 2 · ® S oocnfe-feofeoowfeo o O Ρί fe P o ............

ft O ft G) oooooooooooo o

fairy (OHO

fe fe 3 CD

cd _tn cd fe___ _ fe * Λ SO 4J + 3 + 3 + 3 ^ J4J-J4J-P-Pfefe dJ d \! fi feefefefefefefet — ifefefefe fe • '• fe oøoocooooooo • fe feefefefefefefefefefepDfe!> cm pi feefefefefefefefefefefefef • fe i · η ooooocccoooø fe do fe__Ti Ti Ti_TiTiETEESS_ fe

-J

fe O cn

• fe E

H o

<D CM O

ft, CMS cM ^ nso fe rrL - cfeoEHc-nesocn 1¾

O S-i s CE ^ I! O O fe CM CM CM CM CE CM

-2. o ε ^ ε ιπεοεεεεεε ft osoooooooooo

fe £ '- · O CM CM CM CM CM CM CM CM CM CM CM

s ° E '-' EEEEESSEEE

<3 o ooooooooooo p <J oooooooooooo 1! X É É E ~ “s s S S X X X X ø fe OOOOOOOOOOOO Pi

CfiA, llllllllllll ft o · saaaaasda «« o tn ------------ ri X s SSSES Ξ EE EEE fe ~~ 7j ι-r- »ir- * ι-r-» »-— T -— * * -r- <»µ-im * rl H oooooooooo o 2 or IIIIIIIIIIOI p aaassaaaaaa d fe

Wi-j-4 »- * - *» —-1 l-j-l »* —i» - «Ht-t I — I H-f

►J-, »-i-l» —i HH ι-fe hi- I — I HH M-C

tn · feCMi ^ -ONfel ^ * · On ON NO CM

X U ΙΛ lf \ C \ tfi 1Λ ^ NO fe ·!> O ft E I__________i 7 143905

Furthermore, the new 3-substances are chemically more stable than the fusidic acid itself, for example, the l6-acetoxy group in fusidic acid will under certain conditions be hydrolyzed to a hydroxy group, whereby a significant decrease in activity takes place. In the substances prepared according to the invention, the 16 substituents are not likely to be hydrolyzed. For example, it has been demonstrated that when a 2 # (w / v) solution of the compounds of the invention in aqueous buffer solution of pH 9 6 is maintained at 40 ° C for 30 days, no decomposition products can be detected even by thin layer chromatography.

Like fusidic acid, the new drugs are effectively absorbed from the gastrointestinal tract and are practically non-toxic.

The present invention relates, as mentioned, to an analogous process for the preparation of a compound of formula I. In the present process, a compound of general formula Vb is reacted: H

J: 2V

COOR3 j Vb 52 = ΛΑ '"Υ CHjJ CH, / & [f I CH- i.AAJ 3 ch3 1 1 H__ 2 in which Q» star for Q or a group, R is an alkanoyl-, 2

aralkanoyl or an aroyl group, Q, is as defined above, Y

g represents chloro, bromo or iodo and represents a straight or branched alkyl group having from 1 to 6 carbon atoms, an aralkyl, alkanoyl or aroylmethyl group, an alkanoyl or aroyl oxyalkyl group, an alkyloxymethyl or cyanoethyl group, with a compound of formula VII: R 1 -AH, in which R 1 is as defined above and A represents oxygen or sulfur, thereby forming a compound of formula VIII: 143906 during the rearrangement of the configuration at C-16

J

l ^^ COOR3 jC a-r1 * VI11 iXXT ^ i

Q

cn3 1 2 13 in which Q 1, Q, R, R and the dotted line between C-2b and C-25 are as defined above and A is oxygen or sulfur. When A in Formula VII and VIII stands for oxygen, the reacting compounds of Formula VII may advantageously be used as solvents and the reaction may be carried out in the presence of a silver or mercury salt, e.g., silver carbonate, silver trifluoroacetate or mercury acetate, or a base, e.g., potassium carbonate. , sodium bicarbonate or sodium alcoholate, and at room temperature or slightly elevated temperature.

When A in Formula VII and VIII stands for sulfur, the reaction may be carried out 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 at room temperature or at slightly elevated temperature.

In a final step, the compounds of formula VIII can be converted to compounds of formula I by hydrolysis, for example, in aqueous methanol or ethanol and in the presence of a base such as sodium or potassium hydroxide or carbonate.

1 2

Compounds of formula VIII, in which Q 1 and Q represent the group or oxygen, and represent a readily hydrolyzable x 10 0 ester group corresponding to R, are without further conversion end products prepared according to the invention.

Compounds of formula VIII, in which QX 'and stands for H 3 o the group n. Or oxygen, and RJ represents an unsubstituted or xl for compounds of formula I by a reduction. In the case where R 2 represents a benzyl or cyanomethyl group, a catalytic hydrogenation is preferred, whereas if R represents an acetonyl or phenacyl group, a reduction of zinc in acetic acid would be preferred.

Compounds of formula I in which stands for oxygen and

2 H

Q stands for either oxygen or for the group n 2 ->, can also be prepared by corresponding compounds of formula I, in which Q and 2 j Q represent the group HQ 2>, by oxidation methods well known to those skilled in the art.

1 2

Compounds of formula I in which Q or Q stands for H

7 + * can also be prepared by reducing corresponding for-H0 ~ 1 2 bonding dexterities in which Q and Q represent oxygen.

Compounds of general formula I or VIII in which A represents a sulfinyl group can be prepared by reacting the corresponding compounds of formula I or VIII in which A represents sulfur with an oxidizing agent, e.g. hydrogen peroxide, sodium metaperiodate or chromic acid . The reaction is carried out in an inert solvent, for example water, acetic acid, ethanol, or acetone below or at room temperature or at slightly elevated temperature.

The easily hydrolyzable esters of compounds of formula I can be prepared in known manner by methods described in the literature.

Compounds in which there is a single bond between C-24 and C-25 can also be prepared according to the invention from the corresponding unsaturated compounds by reduction, for example a catalytic hydrogenation, where, for example, palladium-on-carbon is used as catalyst.

In addition, intermediates of formula VIII can be prepared by one or more of the following methods: a) Compounds of general formula VIII in which A represents sulfur and R · · · is an aryl or aromatic heterocyclyl group can be prepared by reacting a compound of the general formula IV, in which the hydroxyl group at C-16 is tt-oriented, with a phosphine, for example, tributylphosphine or triphenylphosphine, and a compound of the general formula R

The reaction is carried out in an inert organic solvent, preferably dimethylformamide or pyridine, and below or at room temperature.

b) In another embodiment of the method, a compound of formula IV is reacted in which the hydroxyl group at C-16 is α-oriented,

1 H

and Q 'is different from the group H0, with a reactive derivative of an alkyl or arylsulfonic acid, for example an acid chloride or anhydride to form a compound of the general formula IX: J3) R5

-OR5 IX

.

CH3

P Q

wherein Q., R-3 and the dotted line between C-24 and C-25 are as defined above for compounds of formula IV, Q1 'represent oxygen or the group R2Q (R is an alkanoyl, aralkanoyl or aroyl group) and R represents an alkyl or an arylsulfonyl group, preferably a methanesulphonyl or a p-toluenesulfonyl group.

In a subsequent step, a compound of formula IX is reacted with a compound of general formula VII, thereby forming a compound of general formula VIII. If A in Formula VII and VIII stands for oxygen, the compounds of Formula VII can be used as solvents and the reaction can be carried out at room temperature or slightly elevated temperature, optionally in the presence of an organic base, eg triethylamine. When A in Formula VII and VIII stands for sulfur, the reaction can be carried out in the same manner as described above for the conversion of substances of Formula Vb into compounds of Formula VIII in which A represents sulfur.

c) In another embodiment, compounds of general formula II in which Q 1, Q, R, the dotted line between C-24 and C-25 and A may be as defined above, and R hydroxy-substituted alkyl group is converted into corresponding compounds in which R "represents a halogen-substituted alkyl group by methods described below for conversions of compounds of formula IV into compounds of formula Va or Vb.

In the 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 base, with an aliphatic or aromatic mercaptan, preferably in the presence of a base, with ammonia or an aliphatic or aromatic amine, or with salts of lower alkanoic or benzoic acid, with silver or sodium fluoride, with alkali metal azides, nitrites, cyanides or thiocyanates, or with salts of lower thioalkanoic acids or thiobenzoic acid, thereby forming compounds of formula VIII, in which H1 represents an alkyl group substituted, for example, with a fluorine atom, an alkyloxy, aralkyloxy, aryloxy, alkylthio, aralkylthio, arylthio, amino, alkylamino, dialkylamino, azido, nitro, cyano, thiocyano -, alkanoyloxy, aralkanoyloxy, aroyloxy, alkanoylthio or aroylthio group.

d) The C-2k, 25 unsaturated compounds may in some cases be advantageously hydrogenated to the corresponding saturated intermediates of formula VIII. The compounds of formula VIII can be transferred to substances of general formula I, as previously described.

The compounds of formula Vb can be prepared by a process in which a first step reactes a compound of formula II with a compound of formula III, thereby forming a compound of general formula IV

H3Sif3 H3 <ViH3 β Έ

I COOX I

Tf '*' "L ^ cooir Q VYii ^ oH + y_r3_>

CH3? Η3 Ί CHI C.hT I

s.XXJ * '.XØX

2 in which formulas Q 1, Q and the dotted line between C-24 and C-25 12 143905 are as defined above, the wavy line of the C-16 hydroxylyl group shows that it may be oc- or β-oriented, X stands for hydrogen or a cation such as Na +, K +, Ag +, an ammonium or trialkyl 3 ammonium ion, Y is chlorine, bromine or iodine and R represents a straight or branched alkyl group having from 1 to 6 carbon atoms, e.g. ethyl or tert-butyl group, a substituted or unsubstituted aralkyl group, for example a benzyl, p-nitrobenzyl or p-methoxybenzyl group, an alkanoylmethyl or aroylmethyl group, for example an acetonyl or phenacyl group, an alkanoyloxyalkyl or aroy for example, an acetoxymethyl, pivaloyloxymethyl or benzoyl oxymethyl group, an alkyloxymethyl group or a cyanomethyl group.

The reaction is carried out in an inert organic solvent, e.g., dimethylformamide, and at room temperature or some elevated temperature.

The preparation of the compounds of formula II is either known from the literature, or is disclosed in U.S. Pat.

4,004,004, or may be prepared by analogous methods.

In the next step, the compounds of formula IV are converted to compounds of formula Va or Vb J3 ΗΠ CH3 OOR ^ L ^^^ COOR · ^

CH2 "CH2_J

/on TV

In j CH 3 Va Vb 12 3 in which formulas Q ', Q, R, Y and the dotted adhesive between C-2h and C-25 are as defined above.

The conversion is carried out by leaving a compound of formula 13 143905

IV react with a polyhalomethane such as tetrachloromethane or tetrabromomethane, or an N-halogenamide such as N-chlorosuccinimide, in the presence of triphenylphosphine, a trialkylphosphine, a triarylphosphite or hexamethylphosphoric triamide, or with an immonium salt of the general formula

(ch ^^ ch-o-r ^ Y ~ VI

4 in which R represents an unsubstituted or substituted phenyl group, a phenyloxycarbonyl group, an alkyl, benzoyl or acetyl group, and Y- is a chloro, bromo or iodo ion. The reaction is carried out in an inert solvent, for example ether, tetrahydrofuran, dimethylformamide, acetonitrile, and at or at room temperature. The compounds of formulas Va and Vb may 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 methylene) dimethyliminium halide.

The conversion of the compounds of general formula IV into compounds of formulas Va and Vb usually causes a rearrangement of the configuration at the C-16 carbon atom. Thus, a compound of formula IV with a cc-oriented hydroxyl group at C-16 will be rearranged to a compound of formula Va, and a compound of formula IV with a β-oriented hydroxyl group at C-16 will be converted to a compound of formula Vb. However, compounds of formula Va can be rearranged to 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 iodide, in suitable solvents, preferably dimethylformamide, acetonitrile or acetonitrile, somewhat higher temperature. When a compound of formula IV, 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-N, N-dimethylformimidate bromide, the first substance of formula Va is rearranged to a substance of formula Vb during the reaction.

The compounds of formula Va and Vb are novel compounds.

The compounds of the present invention can be included in pharmaceutical compositions which can be used in the treatment of infectious diseases in human and veterinary practice.

With this in mind, such compositions as an active component will contain at least one compound of formula I, or a salt thereof with a non-toxic, pharmaceutically acceptable base, or a readily hydrolyzable ester of these compounds, together with solid or liquid pharmaceutical carriers. and / or excipients.

In such compositions, the ratio of therapeutically active material to carrier can vary between 1 and 95% by weight.

The compositions may be processed into various pharmaceutical forms of administration, such as granules, tablets, pills, dragons, suppositories, capsules, delayed-release tablets, suspensions, injections, and, as regards mixtures, they may be filled into bottles or tubes or similar containers.

The pharmaceutical preparation may further contain other pharmaceutically active components which may be advantageously administered with the compounds of the invention for the treatment of infectious diseases, such as other suitable antibiotics, in particular such antibiotics which enhance activity and / or prevent the development of resistance. Such antibiotics include penicillins, cephalosporins, tetracyclines, rifamycins, erythromycin, lincomycin and clindamycin. Among other compounds which may be advantageously combined with the compounds of the invention, especially for topical treatment, may be corticosteroids such as hydrocortisone, triamcinolone or fluocinolone.

When the substances are administered in the form of salts with pharmaceutically acceptable non-toxic bases, the preferred salts are, for example, the easily soluble sodium salts or diethanolamine salts, but other pharmaceutically acceptable non-toxic salts may be used, e.g., salts which are only slightly soluble in water for the purpose of obtaining a special and suitable absorption rate.

The following are a number of examples of preparation of intermediates.

Preparation 1

Phenacyle ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid A.

The sodium salt of 3- ° -acetyl-16-epideacetylfusidic acid (5 * 38 g) and phenacyl bromide (2.2 g) was dissolved in dimethylformamide (40 ml).

After standing for 16 hours at room temperature, the solution was diluted with 150 ml of ether, washed with water (4 x 50 ml), dried, and evaporated in vacuo to isolate 6.2 g of the phenacyl ester of 3-O-acetyl ether. l6-epideacetylfusidic acid as a colorless foamy mass.

U3906 15 B, Phenacylester of 3-O-acetyl-l6-deaeetoxy-l6g-brine fusidic acid A suspension of dimethylformamide (1.1 ml) and phenyl chloroformate (5.04 ml) in 80 ml of petroleum ether (b.p. 50 ° c) was stirred vigorously at room temperature. Over one hour, carbophenoxy-N, N-dimethylformimidate chloride was formed as colorless crystals. This product released carbon dioxide by further stirring for 16 hours to give crystalline phenyl-N, N-dimethylformimidate chloride. This was then converted into N, N-dimethylformamide diphenyl acetal by adding a solution of phenol (3 »76 g) and triethylamine (5» 56 ml) in ether (10 ml) to the reaction mixture with stirring. After further stirring for one hour, the by-product of triethylammonium chloride was filtered off and washed with 50 ml of petroleum ether. By adding acetyl bromide (2.0 ml) with stirring to the combined filtrate and washing liquid, phenyl-N, N-dimethylformimidate bromide was formed as colorless crystals. These were collected and washed with petroleum ether to remove traces of phenyl acetate.

The thus obtained immunomodium bromide (6 g) was very hygroscopic and was immediately added to a solution of the phenacyl ester of 3-O-acetyl-16-epideacetylfusidic acid (6.2 g) in dimethylformamide (cow ml). After standing for 8 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) dry, and evaporated in vacuo. Addition of methanol (50 ml) caused the residue to crystallize. The crystals were filtered off, washed with methanol and dried to isolate 5 »2 g of phenacyl ester of 3-O-acetyl-l6-deacetoxy-l6a-bromfusidic acid, m.p.

Preparation 2-8

Esters of 3-O-acetyl-16-deacetoxy-16a-bromofusidine acid A. Following the procedure of Preparation 1A but replacing the phenacyl bromide used with the esterifying agent shown in Table I, the esters of 3-O-acetyl-16-epideacetylfusidic acid were obtained. shown in Table I.

B. By substituting the phenyl ester of 3-O-acetyl-16-epideacetyl-fusidic acid with the esters of 3-O-acetyl-16-epideacetyl-fusidic acid shown in Table I and otherwise following the procedure described in Preparation 1B, the esters of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid shown in Table I.

Table I: (next page) Η3γΗ3 k / COOR2 '' r1

c hu ch.T T

JjL% in I

jTTTV

CH COO 'J 1 CH3

Resulting compounds Ϊ 2

Preparation Esterifying agent R R melting point _____ (° C) 2 A chloromethyl acetal OH CH 2 COCH 2 amorphous 2 B Br CHgOCOCH ^ 102-105 3 A chloromethyl pivalate OH CHgOCOCiCH 2) ^ amorphous 3 B Br CHOCO (CH) amorphous

£ j J

4 A chloromethyl benzoate OH CH 2 COCgH ^ amorphous 4 B Br CH2 COC6H5 93-9 ^ 5 A chloroacetonitrile OH CH2CN amorphous 5 B Br CH2CN 122-123 6 A benzyl bromide OH CHgCgH ,, 108-109 6 B Br CH ^ gH 128-129 7 A p -methylbenzyl bromide OH CH 2 g H 2 CH 2 amorphous 7 B Br CH 2 Cl 2 H 2 CH 2 amorphous 8 A chloromethyl methyl ether OH CH 2 OCH 2 amorphous 8 B Br CH 2 OCH 3 amorphous

<£ J

Preparation 9 p-Nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid

The sodium salt of 3-O-acetyl-16-epideacetylfusidic acid (21.5 g) and p-nitrobenzyl bromide (9 »5 g) was dissolved in dimethylformamide (200 ml). The solution was allowed to stand at room temperature for 16 hours, forming the p-nitrobenzyl ester of 3-O-acetyl-16-epideacetyl-173,3905 fusidic acid. Then phenyl N, N-dimethylformimidate bromide (36 g; see Preparation IB) was added and the resulting reddish-brown solution was added at room temperature for 48 hours. Methanol (700 mL) and water (80 mL) were added with stirring to precipitate a crystalline product. The crystals were filtered off, washed with methanol: water 3: 1, and dried. 26.1 g of p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16oc- * bromfusidic acid were isolated, mp 151-157 ° C.

Preparation 10-12

Breasts of 3-O-acetyl-16-deacetoxy-16g-bromfusidic acid

Following the procedure of Preparation 9 but replacing p-nitrobenzyl bromide with one of the esterifying agents listed in Table II, the esters of 3-O-acetyl-16-deacetoxy-16oc-bromofusidic acid shown in Table II were obtained .

Table II: h3c ch3

k / C00R

CH COO

J 1 ch3

Resulting compounds melting point

Preparation Esterifier R (° C) 10-p-benzylphenacyl bromide d ^ COC ^ H ^ CH ^ C ^ H ^ 127-129 11 p-methoxyphenacyl bromide CHoC0CgH

Preparation 13

Benzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid A. Benzyl ester of 3-O-acetyl-16-deacetylfusidic acid

To the sodium salt of 16-deacetylfusidic acid (84.7 s) in dimethylformamide (200 µl) was added benzyl bromide (25 ml). After stirring for 5 hours at room temperature, the resulting solution was cooled to 0 ° C and pyridine (200 ml) and acetic anhydride (170 ml) were added. After standing at room temperature for 16 hours, the mixture was again cooled to 0 ° C and 50 ml of water was added with stirring at a rate such that the temperature remained below 15 ° C (about 1 hour). Then, methanol (800 ml) and water (400 ml) were added to complete the precipitate of the desired product, which was filtered off for 1 hour at 10 ° C, washed with ice-cold methanol (3 x 20 ml) and dried to give 68 g of benzyl ester of 3-O-acetyl-16-deacetylfusidic acid as colorless crystals, mp 154-158 ° C.

B. Benzyl Ester of 3-O-Acetyl-16-Deacetoxy-16cx-Bromofusidic Acid The benzyl ester (68 g), sodium bromide (46.2 g), pyridine (22 ml), and dimethylformamide (400 ml) mixed and stirred for 30 minutes. minutes at room temperature and then cooled to 0 ° C. Phenyl chloroformate (56.5 ml) was added over a 45 minute period and the mixture was stirred at room temperature for 18 hours. The reaction product was then precipitated by the addition of methanol (400 ml) and water (300 ml). The colorless crystals were filtered off and washed with methanol: water (2 x 60 ml, ratio 2: 1) and then with petroleum ether (3 x 30 ml). After drying, 62.9 S of benzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid, mp 124-126 ° C, was obtained.

Preparation 14-15

Esters of 3-O-acetyl-16-deacetoxy-16a-bromofusidic acid A. Following the procedure of Preparation 13A but replacing benzyl bromide with phenacyl bromide or p-nitrobenzyl bromide, the esters of 3-O-acetyl-16-deacetylfusidic acid were obtained in Table III.

B. Following the procedure of Preparation 13 B but replacing the benzyl ester with the phenacylester or p-nitrobenzyl ester in Table III, the esters 14B and 15B shown in Table III were obtained.

Table III: 19 143906 H_C CH_ r.

\> ^ COOR

CH 2 CO 3 in CH 3

Resulting compounds

Preparation 2 Melting point (° C)

R R

l4 A β-ΟΗ CH2COC6H5 149-151 14 B α-Br CH2COC6H5 l4l-l42 15 A β-ΟΗ 0Η206Η4Ν02- (ρ) 141-143 15 B a-Br CH ^ gH ^ NO ^ (p) 157-159

Preparation 16

Pivaloyloxymethyl ester of 3-Q-acetyl-16-deacetoxy-16 (X-bromofusidic acid A. Pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16 [3-bromfusidic acid

Pivaloyloxymethyl ester of 3-O-acetyl-16-epideacetylfusidic acid (17.4 g) was dissolved in dry ether (200 ml)} triphenylphospin (16 g) and tetrabromomethane (20 g) was added. After stirring for 16 hours at room temperature, the reaction mixture was filtered to remove triphenylphosphine oxide which formed as a by-product.

The filtrate was evaporated in vacuo and the residue purified by chromatography on silica gel (cyclohexane ethyl acetate 8: 2) to give 10.6 g of pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16β-bromofusidic acid in colorless crystals. ether-petroleum ether having a melting point of 120-122 ° C. Recrystallization of ether-petroleum ether gave an analytically pure product, mp 120-122 ° C.

B. Pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-α6α-bromfusidic acid

The above-described pivaloyloxymethyl ester (5 g) was epimerized to the 16oc compound by leaving it with tetrabutylammonium bromide (5 g) in acetonitrile (60 µl) for three days at room temperature. The reaction mixture was evaporated in vacuo and ether was added to the residue to crystallize tetrabutylammonium bromide. The crystals were filtered off and the filtrate washed with water (2 x 50 ml), dried and evaporated in vacuo to give 4.87 g of pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid as a colorless gum.

Preparation 17

Acetoxymethyl ester of 3-O-acetyl-16-deacetoxy-16a: -bromofusidic acid A. Acetoxymethyl ester of 3-O-acetyl-16-deacetoxy-168-bromo-fusidic acid

Following the procedure of Preparation 16A but replacing the pivaloyloxymethyl ester of 3-O-acetyl-16-epideacetylfusidic acid with the corresponding acetoxymethyl ester, the acetoxy-methyl ester of 3-O-acetyl-16-deacetoxy-63-bromofusidic acid was prepared with m.p. 119-120 ° C.

B. Acetoxymethyl ester of 3-O-acetyl-16-deacetoxy-16a-bromo-fusidic acid

Following the procedure of Preparation 16 B, but replacing the pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16β-bromo-fusidic acid with the corresponding acetoxymethyl ester, the acetoxymethyl ester of 3-O-acetyl-16-deacetoxy-16α-bromofusidine acid, m.p. 102-105 ° C.

Preparation 18

Phenacyl ester of l-O-acetyl-l6-deacetoxy-l6a-bromo-24.25-άϊΙινάΓθ-fusidic acid A. Phenacyl ester of l6-deacetyl-24,25-dihydrofusidic acid

To a solution of the sodium salt of 16-deacetyl-24,25-dihydrofusidic acid (4.99 g) in dimethylformamide (25 ml) was added phenacyl bromide (1,99 g) and the mixture was stirred at room temperature for 4 hours. After dilution with ether (100 ml), the mixture was washed with water (4 x 25 ml). The organic phase was separated, dried and concentrated to ca. 20 ml, whereby a crystalline product precipitated. After standing in a refrigerator for 2 hours, the crystals were filtered off, washed with ether and dried to give 4.52 g of the desired compound, mp 92-94 ° C (probes).

B. Phenacylester of 3-O-Acetyl-16-deacetyl-24,25-dihydrofusidinic acid

To a solution of phenacyl ester of 16-deacetyl ~ 24,25-dihydrofusidic acid (2.38 g) in pyridine (8 ml) was added under stirring acetic anhydride (4 ral) and the mixture was allowed to stand at room temperature for 16 hours. After diluting the reaction mixture with diisopropyl ether (60 ml) with stirring, a colorless compound crystallized which, after washing with diisopropyl ether and drying, afforded 1.92 g of the desired compound, mp 133-135 ° C.

C. Phenacylester of 3-O-acetyl-16-deacetoxy-16 (X-broni-24,25-dihydrofusidic acid

Phenyl chloroformate (1.26 ml) was added to the solution and at 0 ° C was added a stirred solution of the phenyl ester of 3-O-acetyl-16-deacetyl-24,25-dihydrofusidic acid (1.59 g), sodium bromide (1.03 g) ) and pyridine (0.52 ml) in dimethylformamide (15 ml). After the addition was complete (^ 15 min), the reaction mixture was stirred for 2 h at 0 ° C and then at room temperature for 16 h. Dropwise addition of methanol s water (15 ml) with stirring to the mixture precipitated a crystalline product which was filtered off, washed with methanol and dried. There was thus obtained 1.22 g of the desired product which after recrystallization of methylene chloride-methanol gave an analytically pure product, mp 130-132 ° C.

Preparation 19-20

Esters of 3-O-acetyl-16-deacetoxy-16ff-bromo-24,25-dihydrofusidic acid9 A. Following the procedure of Preparation 18A but replacing phenacyl bromide with benzyl bromide or p-nitrobenzyl bromide, esters of 16-deacetyl-24 were obtained. , 25-dihydrofusidic acid shown in Table IV below.

B. Following the procedure of Preparation 18 B but substituting the phenacyl ester of 16-deacetyl-24,25-dihydrofusidic acid with corresponding esters shown in Table XV, esters of the 3-O-acetyl compounds shown in Table XV were prepared.

C. By following the procedure of Preparation 18 C, but substituting phenyl ester of 3-O-acetyl-16-deacetyl-24,25-dihydro-fusidic acid with the corresponding esters shown in Table IV, esters of 3-O-acetyl-16 were prepared. -deacetoxyl-l6a-bromo-24,25-dihydro-fusidic acid shown in Table IV.

Table IV; Η3 <^ ν / Η3 I ^ JSOOR3 Η0'άλ ^ η2

chJ Ch3_J

.v-Cu1 ^ k

Resulting compounds 12 3 o

Preparation RR R-5 Melting Point (C) 19 A Η β-ΟΗ CH 2 C 6 H 5 amorphous 19 B CH-CO β-ΟΗ CH-C ^ H- 162-163 J ^ 05 19 C CH₂CO0-Br CH₂H-104 -105 jo 5 20 A Η β-ΟΗ CHgCgH ^ NOg-p amorphous 20 B CH ^ CO β-ΟΗ 0Η206Η ^ Ν02-ρ amorphous 20 C CH ^ CO 'α-Br ΟΗ ^ Η ^ ΝΟ ,, - ρ l47- l49 ί

Preparation 21

Benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16q-bromfusidic acid A. Benzoyloxymethyl ester of 16-epideacetylfusidic acid 16-Epideacetylfusidic acid (35 »5 phenolphthalein as indicator. After evaporation to dryness in vacuo, the amorphous sodium salt was dissolved in dimethylformamide (150 ml), added with chloromethyl benzoate 2 µg (14.08 g), and the mixture was stirred at room temperature for 16 hours. Water (200 mL) was added and the mixture extracted with ether (CO mL). The organic phase was separated, washed with water (k x 100 ml), dried and evaporated to dryness in vacuo to give hp, gg of benzoyloxymethyl ester of 16-epideV acetylfusidic acid as an amorphous product.

B. Benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16g-bromfusidic acid -

The benzoyloxymethyl ester prepared above was dissolved in dime thylformamide (300 ml); phenyl-N, N-dimethyl-r formimidate bromide (67 g) was added with stirring and the reddish brown solution was left at 5 ° C for 6-7 days. To the mixture was then added methanol (150 ml) and with vigorous stirring water (ISO ml) from a separatory funnel whereby a crystalline product precipitated. The crystals were filtered off, washed with methanol: water 1: 1, pg dried to give 27.1 g of benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16a-bromfusidic acid, mp 131-135 ° C. Two recrystallizations of ether-methanol raised the melting point to 140 ° -42 ° C.

Preparation 2Z-2k

Esters of 3-O-formyl-16-deacetoxy-16a-bromfusidic acid A. Following the procedure of Preparation 21A, but replacing chloromethylbenzoate with benzyl bromide, chloromethyl pivalate or chloromethyl acetate, the esters of 16-epideacetylfusidic acid were prepared in Table V.

B. Following the procedure of Preparation 21 B, but replacing the benzoyloxymethyl ester of 16-epideacetylfusidic acid with the other corresponding esters shown in Table V, the esters of 3-O-formylar 16-deacetoxy-16ct-bromfusidic acid shown in Table V. were obtained.

TT p ρττ

Table V: (next page) 3 L ^ -COOR ^

«4 <rHJ_J

, Ref V ·. 143905 24

Resulting compounds

Preparation pi p2 p3 melting point (° C) 22 A H OH CH 2 C 6 H 5 95-98 22 B HCO Br CH 2 CgH 2 125-127 23 A H OH CH OCOC (CH) amorphous

J ^ J J

23 B 1 HCO Br CHgOCOCiCH ^ amorphous 2k A H OH CH ^ COCH ^ amorphous 2k B HCO Br CH2OCOCH3 123-125

Preparation 25

Benzyl ester of 3-O-formyl-16-deacetoxy-16g-bromfusidic acid A. Benzyl ester of 16-deacetylfusidic acid

To a solution of the sodium salt of 16-deacetylfusidic acid (1, 97 g) in dimethylformamide (25 ml) was added benzyl bromide (1.5 ml) and the mixture was stirred at room temperature for one hour. 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 was dissolved in ether (50 ml) and by the addition of petroleum ether (50 ml) with stirring a crystalline product precipitated. The crystals were filtered off, washed with ether: petroleum ether 1: 2 and dried to give 4.92 g of the desired product, mp 117-119 ° C.

B. Benzyl ester of 3-O-formyl-16-deacetylfusidic acid

To a solution of benzyl ester of 16-deacetylfusidic acid (4.52 g) in pyridine (8 ml) was added dropwise at 0 ° C to 4 ml of the mixed anhydride of acetic acid and formic acid. The mixture was kept at the low temperature for 15 minutes. With stirring, the reaction mixture was diluted with diisopropyl ether (4θ ml) to precipitate a crystalline product. After two hours in the refrigerator, the crystals were collected, washed with diisopropyl ether and dried to give 4.04 g of the desired product, mp 433-145 ° C. Recrystallization of ether-diisopropyl ether gave an analytically pure product, mp 145-147 ° C.

Benzyl ester of 3-O-formyl-16-deacetoxy-16 (X-bromofusidic acid)

Following the procedure of Preparation 13B, replacing the benzyl ester of 3-O-acetyl-16-deacetyl-fusidic acid with benzyl ester of 3-O-formyl-16-deacetyl-fusidic acid, benzyl ester of 3-O-formyl-16-deacetoxy-16-oct. bromfusidic acid as colorless crystals, mp 125-127 ° C.

Preparation 26

Pivaloyloxymethyl ester of 3-O-formyl-16-deacetoxy ~ 16g-bromo-fusidic acid A. Pivaloyloxymethyl ester of 16-deacetylfusidic acid

To one. solution of the amorphous silver salt of 16-deacotyl-fusidic acid (5 »8 g) in dimethylformamide (50 ml) was added chlorine thylpivalate (1.48 ml) and the mixture was stirred at room temperature for 48 hours. Insoluble material was removed with filter aid, and after washing the precipitate with ether (2 x 25 ml), ether and filtrate were combined and diluted with ether (100 ml) and the mixture washed with water (4 x 50 ml). The organic layer was dried and evaporated in vacuo to give the crude ester as a yellowish foam. It was purified by chromatography on silica gel (solvent: cyclohexane: ethyl acetate 3.7) * and the desired ester was isolated as an amorphous product.

The NMR spectrum (CDCl3) showed signals at 0 = 0.90 (d, 3H), 0.93 (s, 3H), 0.98 (s, 3H), 1.22 (s, 9H; CfCH 1.38 (s, 3H), 1.62 and 1.68 (2 bs, 6H), 2.99 (m, 1H; CH-13), 3.77 (m, 1H; CH-3), 4 , 33 (m, 1H; CH-11), 5.00 (m, 1H; CH-16), 5.12 (m, 1H; CH-24), ° g 5.15 and 5.42 (2 d , J = 7, 2H; OCH 2 O) ppm. Tetramethylsilane was used as internal reference.

B. Pivaloyloxymethyl ester of 3-O-

C. Pivaloyloxymethyl ester of 3-O-formyl-16-deacetoxy-16a-bromfusidic acid

Following the procedure of Preparation 13B but replacing the benzyl ester of 3-O-acetyl-16-deacetylfusidic acid with the pivaloyl-oxymethyl ester of 3-O-formyl-16-deacetylfusidic acid 26 143905, pivaloyloxymethyl ester of 3-O-formyl-16-deacetoxy was obtained. -6a-Bromfusidic acid as a colorless foam.

The NMR spectrum showed signals at δ = 0.78 (s, 3H), 0.87 (d, J = 7, 3H), 1.00 (s, 3H); 1.23 (s, 9H, c (CH 3) 3), 1.47 (s, 3H), 1, 6l and 1.68 (2 bs, 6H), 3.45 (m, 1H; CH-13) 4.35 (m, 1H; CH-11), 5.08 (m, 1H; CH-3), 5.12 (m, 1H, CH-24), 5.62 (bt, 1H, CH 16), 5.82 and 5.92 (2 d, J = 6, 2H; OCH 2 O), and 8.15 (bs, 1H, HCO) ppm. Tetra-methylsilane was used as internal reference.

Preparation 27

Acetoxymethyl ester of 3-O-formyl-16-deacetoxy-16 (X-bromo-24,25-dihydrofusidic acid A. Acetoxymethyl ester of 16-epideacetyl-24,25-dihydrofusidic acid For a solution of the potassium salt of 16-epideacetyl-24,25- dihydrofusidic acid (20.6 g) in dimethylformamide (160 ml) was added chloromethyl acetate (4.0 ml) and the mixture was stirred at room temperature for 18 hours. After dilution with ether (500 ml) the mixture was washed with water (2 x 150 ml) ) and the organic phase was dried and evaporated in vacuo to give the desired product as a colorless foam.

B. Acetoxymethyl ester of 3-O-formyl-16-deacetoxy-16a-bromo-24,25-dihydrofusidic acid

To a solution of the above-prepared ester and sodium bromide (20.6 g) in dimethylformamide (200 ml) was added dropwise at 0 ° C to phenyl chloroformate (25.2 ml).

After the addition was complete (about 45 minutes), the mixture was stirred at 0 ° C for 3-4 hours and then at room temperature for another 10-12 hours. Precipitated sodium chloride was filtered off and washed with dimethylformamide (2 x 25 ml). To the combined filtrate and wash liquid was added methanol: water 1: 1 (300 ml) with stirring to precipitate a crystalline product. The crystals were filtered off, washed with methanol: water 1: 1, dried, and finally recrystallized from ether-diisopropyl ether to give 15.35 S of the desired product, mp 126-127 ° C.

Preparation 28-32

Esters of l6-deacetoxy-l6Q [: - bromfusidic acid

Following the procedure of Preparation 16, but replacing the pivaloyloxymethyl ester of 3-O-acetyl-16-epideacetylfusidic acid with benzyl, phenacyl, pivaloyloxymethyl, acetoxymethyl or benzoyloxymethyl ester of 16-epdeacetylcetic acid, l6a-Bromfusidic acid shown in Table VI.

H C CH

Table VI;

X ^ -COOR

hox Pea c «3

Resulting compounds

Preparation R melting point (° C) 28 CH 2 C / -H_. amorphous 2 o 5 29 CH 2 COC 3 H-amorphous 2 o 5 30 CH 2 OCOC (CH 3) 3 amorphous 31 CH 2 OCO 3

Preparation 33

Phenacylester of 3-O-acetyl-11-keto-16-deacetoxy-16a-bromofusidic acid

To a solution of phenyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid (6.98 g) in acetone (70 ml) was added Jones reagent (3.0 ml) and the mixture was stirred for 30 minutes at room temperature. After dilution with ether (100 ml) and addition of water (70 ml), the mixture was further stirred for 10 minutes.

The organic layer was separated and the aqueous layer was again extracted with ether (100 ml). The combined organic extracts were washed with water for neutral reaction, dried and concentrated to ca. 50 ml to precipitate a colorless crystalline substance. After being refrigerated for 1 hour, the crystals were filtered off, washed with ice-cold ether and dried to yield 5.37 g of phenacylester of 3-O-acetyl-11-keto-16-deaootoxy-16a-bromfusidic acid, m.p. 120-121 ° C, At concentration 28 143905 of the mother liquor, an additional 0.95 g of the desired substance was obtained, m.p. 14-146 ° C. Recrystallization of methylene chloride diisopropyl ether gave an analytically pure substance, mp 120-121 ° C.

Preparation

Benzyl ester of 3-O-acetyl-11-keto-16-deacetoxy-16a-bromofusidic acid Following the procedure of Preparation 33, but replacing the phenyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid with the corresponding benzyl ester, the benzyl ester of 3-O-acetyl-H-keto-16-deacetoxy-16a-bromfusidic acid as a colorless foam. The NMR spectrum (CDCl3) showed signals at δ = 1.00 (s, 3H), 1.05 (s, 3H), 1.02 (d, 3H), 1.27 (s, 3H), 1 , 62 and 1.68 (2 bs, 6h), 2.06 (s, 3H; CH 2 CO), 3.30 (m, 1H; CH-13), 4-, 95 (m, 1H, CH 3), 5.05 (m, 1H, CH-24), 5.22 (s, 2H; CHgCgH ^, 5.60 (bt, 1H; CH-16), and 7.35 (s, 5H; arom Tetramethylsilane was used as internal reference.

Preparation 35

Acetoxymethyl ester of 3-O-formyl-11-keto-16-deacetoxy-16a-bromo-24,25-dihydrofusidic acid

Following the procedure of Preparation 33 but replacing phenyl ester of 3-O-acetyl-16-deacetoxy-16oc-bromfusidic acid with acetoxymethyl ester of 3-O-formyl-16-deacetoxy-6oc-bromo-24,25-dibydrofusidic acid, acetoxymethyl ester was obtained. of 3-O-formyl-11-keto-16-deacetoxy-16a-bromo-24,25-dihydrofusidic acid as a colorless foam.

The NMR spectrum (CDCl3) showed signals at <f = 0.87 (d, J = 5.5 6h), 1.02 (s, 3H), 1.04 (s, 3H), 1.25 (s , 3H), 2.12 (s, 3H; ch3co), 3.35 (m, 1H; CH-13), 5.10 (m, 1H; CH-3), 5.68 (bt, 1H; CH -16), 5.81 and 5.90 (2 d, J = 5.5, 2Η; OCH 2 O), and 8.15 (bs, 1Η; HCO) ppm. Tetramethylsilane was used as internal reference.

Preparation 36

Acetoxymethyl ester of 3-keto-16-deacetoxy-16 (X-bromofusidic acid A. Acetoxymethyl ester of 3-keto-16-epideacetylfusidic acid

To a solution of the potassium salt of 3-keto-16-epideacetyl-fusidic acid (3.06 g) in dimethylformamide (30 ml) was added chlorine thylacetate (0.1 ml) and the mixture was stirred at room temperature for 18 hours. The reaction mixture was diluted with ether (100 ml) and washed with water (4 x 30 ml). The organic phase was separated 29, dried and evaporated in vacuo to give 3 »2 g of the desired product as a colorless foam.

1Λ 3 9 ο ε B. Acetoxymethyl ester of 3-keto-16-deacetoxy-16g-bromfusidic acid Following the procedure of Preparation 27 B, but replacing the acetoxymethyl ester of 16-epideacetyl-24,25-dihydrofusidic acid with the above-prepared acetoxymethyl ester of 3- keto-lé-epi-deacetylfusidic acid, acetoxymethyl ester of 3-keto-l6-deacetoxy-l6a-bromfusidic acid was obtained, m.p. 444-145 ° C.

Preparation 37

Methoxymethyl ester of 3-O-acetyl-16-deacetoxy-16g-chlorfusidic acid

Methoxymethyl ester of 3-O-acetyl-16-epideacetylfusidic acid (1.4 g), triphenylphosphine (2.6 g) and N-chlorosuccinimide (1.3 g) was dissolved in dry ether. After standing for one hour at 35 ° C, precipitated triphenylphosphine oxide was filtered off and the filtrate was evaporated in vacuo. The residue was purified by chromatography on silica gel (cyclohexane: ethyl acetate 70) to give 1.14 g of methoxymethyl ester of 3-O-acetyl-16-deacetoxy-16β-chlorofusidic acid. After recrystallization of cyclohexane, the pure product was obtained, m.p. l49-151 ° C.

Preparation 38

Benzyl ester of 3-O-acetyl-16-deacetoxy-1,60-chlorofusidic acid

Carbophenoxy-N, N-dimethylformimide chloride was prepared by adding phenyl chloroformate (θ, 3 ml) to N, N-dimethylformamide (15 ml). To this solution was added benzyl ester of 3-O-acetyl-16-epideaethyl ethyl fusidic acid (500 mg). After standing for 16 hours at room temperature, the reaction mixture was diluted with ether (100 ml), washed with 2N sodium hydroxide (25 ml) and water (3 x 25 ml), dried and evaporated in vacuo. There was thus obtained 480 mg of benzyl ester of 3-O-acetyl-16-deacetoxy-16β-chlorfusidic acid, which crystallized from ether-petroleum ether, mp 163-165 ° C. Recrystallization of ethyl acetate-petroleum ether afforded the analytically pure product, m.p. l65-166 ° C.

Preparation 39

Benzyl Ester of 3-O-Acetyl-16-Deacetoxy-16q-Chlorfusidic Acid 143905

To an ice-cold solution of benzyl ester of 3-O-acetyl-16-deacetylfusidic acid (1.36 g) in dimethylformamide (10 ml) and pyridine (0.44 ml) was added phenyl chloroformate (1.13 ml) with stirring. of 30 minutes. After stirring at room temperature for 16 hours, the resulting solution was diluted with ether (100 ml), washed with 2N sodium hydroxide (25 ml) and water (3 x 50 ml), dried and evaporated in vacuo. The residue was dissolved in ether (10 ml) and petroleum ether was added to precipitate the reaction product as colorless crystals.

These were filtered off, washed with petroleum ether and dried, and benzyl ester of 3 ° O-acetyl-16-deacetoxy-160C-chlorfusidic acid, mp 115-117 ° C, was isolated. Recrystallization of ethyl acetate-petroleum ether raised the melting point to 120-122 ° C.

The invention will be further described in the following examples.

Example 1 16-Deacetoxy-16B-isopropylthiofusidic acid A. p-Nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16g-isopropylthiophosidic acid p-Nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-6a (28.6 g) was added to a solution of potassium hydroxide (10 g, 85 µg) and isopropylmercaptan (30 ml) in ethanol (100 ml) and the suspension was stirred for 4 days. Then, 500 ml of water was added to complete the precipitation of the desired product. The crystals were filtered off, washed with water: ethanol (1: 2) and dried. 21.5 S of crude p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16β-isopropylthiofusidic acid were isolated, mp 157-166 ° C.

B. l6-Peacetoxy-l68-isopropylthiofusidic acid

A suspension of the above-prepared p-nitrobenzyl ester in a mixture of ethanol (800 ml) and 2N 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 with 5 S charcoal for 15 minutes while still warm.

After filtration, 500 ml of water was added and, after cooling to room temperature, the precipitated crystalline product was filtered off, washed with water and dried to isolate 14.1 g of l6-deacetoxy-l6β-isopropylthiofusidic acid with melting point

31 U390B

223 - 229 ° C · Recrystallization of 2-butanone gave an analytically pure product, mp 229-231 ° C.

Examples 2-10 16β-Thioethers of 1,6-deacetoxyphusidic acid A. p-Nitrobenzyl ester of ΐββ-thioethers of 3-O-acetyl-16-dioxide toxyfusidic acid

Following the procedure of Example 1A but replacing the isopropylmercaptan with the mercaptans shown in Table VII, a series of p-nitrobenzyl esters of ΐ6β-tbioethers of 3-O-acetyl-16-deacetoxy fusidic acid shown in Table VII were prepared.

Table VII:. H ^ C CH ^ cooch2- ^ 0 ^ -no2

^ «S_I

rfjV

i t CH3

Resulting compounds melting point

Example Mercaptan R (C) 2 A Ethylerercaptan CHgCH3 3 A 2-hydroxyethyl CH2CH20H 192-194 mercaptan 4A 2-aminoethyl CHgCHgNHg 188-191 mercaptan 5 A allyl mercaptan CH2CH = CH2 167-170 6 A isobutyl mercaptan CH2CH (CH3) 2 104 -112 7 A sec-butyl mercaptan CH 2 CH 2 JCH 2 CH 3 150-157 8 A cyclopentyl mercaptan cyclopentyl 100-109 9 A mercaptoacetic acid CH COOCH 125-129

methyl ester J

10 A furfuryl mercaptan furfuryl l46-l48 B. l66-Thioethers of l6-deacetoxyfusidic acid

Following the procedure of Example 1B, but substituting p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16P-isopropyl-thiofusidic acid with the p-nitrobenzyl ester of the Ιόβ-ΐΙιΐοβΙΙιβΓρ 32 143905 of 3-O-acetyl-16 deacetoxy fusidic acid shown in Table VIX, the l6P thioethers of the l6 deacetoxy fusidic acid were shown in Table VIII.

Table VIII; H3 \ ^ H3

k ^ -COOH

h% ^ nJL-sr _I

Γ J ^ H3 ho 'CH3

Resulting connections_

Example R Melting point (° c) 2 B CH2 CH3 195-198 3 B CH2CH20H 179-182 4 B CH2CH2NH2 241-250 (decomposition ·) 5 B CH2CH = CH2 196-199 6 B CH2CH (CH3) 2 199-202 7 B CH (CH3) CH2CH3 218-222 8 B cyclopentyl 217-223 9 B CH2C00H 199-202 10 B furfuryl amorphous NMR spectrum (CDC13) of the compound of Example 10B showed signals at / = 0.97 (s, 6h), 1, 32 (s, 3H), 1.60 and 1.68 (2 bs, 6H), 3.00 (m, 1H; CH-13), 3.73 (m, 1H; CH-3), 3.78 (bs, 2H; SCHg), 4.22 (d, 1H; CH-I6), 4.30 (m, 1H; CH-11), 5.10 (m, 1H; CH-24), 6, Ιό , 4 (m, 2H; arom. CH) and 7.32 (bs, 1H; arom. CH) ppm. Tetra-methylsilane was used as internal reference.

Example 11 16-Deacetoxy-168-isopropylthio-24.25-dihydrofusidic acid A. p-Nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-168-iso-propylthio-24,25-dihydrofusidic acid

Following the procedure of Example 1A, but replacing the 33-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromofusidic acid with the p-nitro-benzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromo -24,25-dihydrofusidic acid, the p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16β-isopropyl thio-24,25-dihydrofusidic acid was prepared in the form of colorless crystals, mp 113-H6 ° C.

B. 16-Peacetoxy-168-isopropylthio-24,25-dihydrofusilic acid

Following the procedure of Example 1B, the p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16 (> - isopropyl-thiofusidic acid) is replaced by the p-nitrobenzylostore of 3-O-acetyl-16-deacot-oxy-16P -isopropylthio-24,25-dihydrofusidic acid, 16-deacetoxy-16β-isopropylthio-24,25-dihydrofusidic acid was prepared, m.p. 232-234 ° C.

Example 12 16-Deacotoxy-16β-cyclohexylthiofusidic acid

To a solution of potassium hydroxide (KOO mg; 85% pure) and cyclohexyl mercaptan (2 ml) in ethanol (100 ml) was added p-nitro-benzyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid (1 43 g) and the resulting solution was allowed to stand at room temperature for 5 days. Then, the reaction mixture was diluted with 150 ml of ether, washed with water (3 x 75 ml), dried and evaporated in vacuo. The residual oil, which contained crude p-nitro-benzyl ester of 3-O-acetyl-16-deacetoxy-163-cyclohexylthiofusidic acid, was dissolved in ethanol (80 ml) and 20 ml of a 2 N aqueous sodium hydroxide was added. After stirring for three hours at 60 ° C, 100 ml of water was added and the dark solution was acidified by the addition of 4N hydrochloric acid (15 ml) and extracted twice with ether. The combined organic phases were washed with water (3 x 50 ml), dried and evaporated. The oily residue was purified by chromatography on silica gel (ether: petroleum ether: acetic acid 70: 30: 0.5) to isolate 16-deacetoxy-16 (S-cyclohexylthiofusidic acid by crystallization of ether-petroleum ether, mp 215-220 ° C Recrystallization of ethyl acetate-petroleum ether gave the analytically pure product, m.p. 216-220 ° C.

Examples 13-15

Following the procedure of Example 12, replacing cyclohexyl mercaptan with the mercaptans shown in Table IX, the 16-thioethers of 16-deacetoxyfusidic acid shown in Table IX were prepared.

, 143905 34

Table IX; H Q ^ CiH

ϊ

k ^ -COOH

CR> A ^ OR

ck [CH3 I

Ho '' '^ vrx ^ in CH3

Resulting compounds

Example Mercaptan R Melting point (° C) 13 2-phenylethyl CH 2 CH C, H 208-214 mercaptan 14 n-butyl mercaptan 0Η "0Η_0Η" 0Η "105-118 (decomposition) 15 methyl mercaptan CH2 amorphous NMR spectrum ( CD1 OD) of the compound of Example 15 showed signals at S = 0.89 (d, J = 6, 3H), 1.00 (s, 3H), 1.03 (s, 3H), 1.38 (s , 3H), 1.62 (bs, 6h), 2.13 (s, 3HJ SCH), 3.03 (m, 1H; CH-13), 3.67 (m, 1H; CH-3), 4 , 03 (d, J = 9, 1H} CH-16), 4.26 (m, 1H; CH-11) and 5.10 (m, 1H; CH-24) ppm. Tetramethylsilane was used as internal reference.

Example 16 16-Deacetoxy-163-ethylthiofusidic acid

To a solution of ethyl mercaptan (2.5 ml) in dimethylformamide (10 ml) was added sodium hydride (650 mg of a 55% suspension in oil). After hydrogen evolution had ceased, pivaloyloxy methyl ester of 3-O-acetyl-16-deacetoxy-16oc-bromfusidic acid (750 mg) was added. After standing for one hour at room temperature, the reaction mixture was diluted with ethyl acetate (50 mL) and extracted with 1N hydrochloric acid (25 mL) and water (2 x 25 mL). The organic phase was dried, filtered and evaporated in vacuo. The crude product thus obtained was purified by chromatography on silica gel (ether: petroleum ether: acetic acid kO: 60: 0.5) to give pure 3-O-acetyl-16-deacetoxy-163-ethylthiofusidic acid as a colorless gum dissolved in a mixture of ethanol (20 ml) and 2N aqueous sodium hydroxide (5 ml) and left at 75 ° C for two hours. The reaction mixture was acidified with 1 N hydrochloric acid (15 ml) and extracted with ethyl acetate (50 ml). The organic phase was washed twice with water (20 ml), dried, and evaporated in vacuo to an oil which crystallized from ether-petroleum ether to give 16-deacetoxy-16β-ethylthiofusidic acid as colorless crystals, mp 195-198 ° C.

Examples 17-20 16-β-Thioethers of 16-deacetoxyphusidic acid

Following the procedure of Example 16, but replacing ethyl mercaptan with the mercaptans shown in Table X, the Ιββ-thioethers of 16-deacetoxyfusidic acid shown in Table X were prepared.

Table X: Η3 <"^^ '! Η3

k ^ COOH H0 \ / \> '\ ^ SR

w 1T

ho ^ ch3 __Results_

Example Mercaptan R Melting point (° C) 17 n-propyl mercaptan CH 2 CH 2 CH 2 amorph 18 t, butyl mercaptan 0 (011 6) 200-203 19 phenyl mercaptan Cg H 2 amorphous benzyl mercaptan CH of the compound of Example 17 signals at 6 = 0.90 (d, 3H), 0.99 (s, 6h), 1.37 (s, 3H), 1.62 and 1.66 (2 bs, 6η) , 2.58 (m, 2H; CH 2 S), 3.00 (m, 1H; CH-13), 3.67 (m, 1H; CH -3), 4.11 (d, 1H; CH-16) , 4.24 (m, 1H; CH-11) and 5.12 (m, 1H; CH-24) ppm. Tetramethylsilane was used as internal reference.

The NMR spectrum (CDCl3) of the compound of Example 19 showed signals at δ = 0.95 (s, 6h), 1.10 (s, 3H), 1.35 (s, 3H), 1.60 and 1. 65 (2 bs, 6H), 3.10 (m, 1H; CH-13), 3.74 (m, 1H; CH-3), 4.30 (m, 1H; CH-11), 4.77 (d, 1H; CH-16), 5.11 (m, 1H; CH-24) and 7.0- 7.4 (5H; arom. CH) ppm. Tetramethylsilane was used as internal reference.

The NMR spectrum (CDCl3) of the compound of Example 20 showed signals at <f = 0.97 (s, 6H), 1.36 (s, 3H), 1.62 and 1.66 (2 bs, 6H ), 3.06 (m, 1H; GH-13), 3.66 (m, 1H; CH-3), 3.74 (bs, 2H; SCHg), 4.08 (d, 1H; CH-16) ), 4.24 (m, 1H; CH-II), 5, 14 (m, 1H; CH-24), and 7.3 (bs, 5H; arom. CH) ppm. Tetramethylsilane was used as internal reference.

Example 21 16-Deacetoxy-16B- (1-methyltetrazol-5-ylthio) fusidic acid

A solution of benzoyloxymethyl ester of 16-epideacetyl-fusidic acid (2.2 g) and di (1-methyltetrazol-5-yl) disulfide (1.5 s) in dry pyridine (20 ml) was cooled in an ice bath and tributylph was added. phosphine (1.4 ml). After standing for 18 hours at room temperature, water (200 ml) and ether (400 ml) were added. The organic phase was separated, washed twice with 1N hydrochloric acid and twice with water, dried and evaporated in vacuo. The residue was dissolved in methanol (50 ml) and potassium carbonate (2.4 g) was added. After stirring for 18 hours at room temperature, the solution was acidified with 4N hydrochloric acid (8 ml) and water (200 ml) and ether (100 ml) were added. The organic phase was separated, washed twice with water, dried and evaporated to give 1.84 g of a crude product which was purified by chromatography on silica gel (ether: acetic acid; 100: 0.5) to give .

800 mg of l6-deacetoxy-16β- (1-methyltetrazol-5-ylthio) fusidic acid as a colorless foam.

The NMR spectrum (CDCl3) showed signals at df = 1.00 (s, 3H), 1.06 (s, 3H), 1.4o (s, 3H), 1.62 and 1.68 (2 bs, 6H). ), 3.17 (m, 1H; CH-13), 3.75 (m, 1H; CH-3), 3.87 (s, 3H; 1'-CH 3), 4.37 (m, 1H, CH-11) and 5.42 (m, 1H; CH-16) ppm. Tetramethylsilane was used as internal reference.

Example 22 16-Deacetoxy-168- (2,5-dichlorophenylthio) fusidic acid

A solution of methoxymethyl ester of 3-O-acetyl-16-epideacetylfusidic acid (490 mg) and di (2,5-dichlorophenyl) disulfide (1.07 g) in dry pyridine (4 ml) was cooled to 0 ° C and tributylphosphine ( 0.72 ml) was added. The resulting solution was allowed to stand at 5 ° C for three days. It was then diluted with ether (100 ml), washed with 4N hydrochloric acid, 2N 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 left at 60 ° C for 1 hour. The reaction mixture was acidified with 4N hydrochloric acid (5 ml) and ether (.100 ml) and water (200 ml) were added. The organic phase was separated, washed twice with water, dried and evaporated in vacuo. The residue crystallized from ether-petroleum ether to give 16-deacetoxy-16f ;-( 2,5-dichlorophenylthiojfusidic acid, m.p. 16-164 ° C.

Example 23 l6-Peacetoxy-l66- (2-azidoethylthio) fusidic acid in A. p-Nitrobenzyl ester of 3-O-acetyl-l6-deacetoxy-l6 [1- (2-bromoethyl thio) fusidic acid

To a solution of p-nitrobenzyl ester of 3-O-acotyl-16-deacetoxy-16β- (2-hydroxyethylthio) fusidic acid (1 g) in 50 ml of dimethylformamide was added 3 g of phenyl-N, N-dimethylformimidate bromide.

After standing at room temperature for 18 hours, the reaction mixture was diluted with ether (50 mL), washed with 2N sodium hydroxide (20 mL) and water (3 x 50 mL), dried and evaporated in vacuo. Addition of ether-petroleum ether to the residue caused it to crystallize. After filtration, washing with petroleum ether and drying, 800 mg of the p-nitrobenzyl ester of 3-O-acetyl-l6-deacetoxy-l6 (1- (2-bromomethyl thio) fusidic acid) was melted to m.p.148-150 ° C.

B. l6-Deacetoxy-l6B- (2-azidoethylthio) fusidic acid t

The 2-bromomethylthioether prepared above was dissolved in 25 ml of dimethylformamide. Lithium azide (400 mg) was added and the reaction mixture was left at 20 ° C for 24 hours. 100 ml of ether was 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 2N aqueous sodium hydroxide. After standing at 60 ° C for 3 hours, the solution was acidified with 8 ml of 4N hydrochloric acid and water (100 ml) and ether (100 ml) were added. The organic phase was separated, washed with water (4 x 50 ml), dried and evaporated in vacuo. Adding ether and petroleum ether to the residue caused l6-deacetoxy-16β- (2'-azidoethylthio) fusidic acid to precipitate as colorless crystals which, after filtration, washing with petroleum ether and drying, afforded 140 mg, mp 173-170 ° C.

Example 24

Sodium salt of l6-deacetoxy-l63- (2-methoxyethylthio) fusidic acid For a solution of p-nitrobenzyl ester of 3-O-acetyl-l6- (ϊθηοβΐο; χ3Γ-ΐββ- (2-bromethylthio) fusidic acid (Ig) A) in methanol (50 ml) was added silver carbonate (1g) and the mixture was stirred at room temperature for 16 hours. Insoluble material was filtered off and washed with methanol (10 ml). The combined filtrate and washing liquid were evaporated in vacuo and the residue 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) and water (2OO ml) was added. and the ether (200 ml) The organic phase was separated, washed twice with water, dried and evaporated in vacuo. The residue was purified by chromatography on silica gel (ether: acetic acid, 100: 0.5) to give the desired product as This was converted to the crystalline sodium salt by dissolving in methanol (25 ml), titrating with 2N aqueous sodium umhydroxide, evaporation and addition of acetone. The crystals were filtered off, washed with acetone and dried.

The NMR spectrum (CD 1 OD) showed signals at S = 1.00 (s, 6h), 1.36 (s, 3H), 1.62 (bs, 6H) 2.78 (2H; Cg 2 S), 51 (2H; Cg 2 O), 3.68 (m, 1H; Cg-3), 4.10 (d, 1H; CH-16), 4.21 (m, 1H; CH-11) and 5.11 ( m, 1H; CH-24) ppm. Tetramethylsilane was used as internal reference.

Example 25 16-Peacetoxy-163- (2-isopropylthioethylthio) fusidic acid

To a solution of potassium hydroxide (500 mg) and isopropyl mercaptan (1.5 nil) in ethanol (50 ml) was added p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-163- (2-bromomethylthio) fusidic acid (lg ) (see Example 23A) and the mixture was stirred for 16 hours at room temperature. Yand (100 mL) and ether (75 mL) were added, the organic phase was separated, washed with 2N 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 (100 ml) and 2N sodium hydroxide (20 ml) and the solution was stirred for 3 hours at 60 ° C. Then 4N hydrochloric acid (15 ml), water (250 ml) and ether (100 ml) were added. The organic phase was separated, washed with water (2 x 50 ml), dried and evaporated in vacuo. The residue 39

U390S

was purified by chromatography on silica gel (ether: petroleum ether: acetic acid, 70: 30: 0.5) to give 400 mg of l6-deacetoxy-16R- (7-isopropyl thioethylthio) fusidic acid.

The NMR spectrum (CDCl3) showed signals at ¢ / = 0.96 (bs, 6h), 1.22 (d, j = 7, 6h), 1.33 (s, 3H), 1.58 and 1, 67 (2 bs, 6H), 2.73 (bs, 4H; SCH 2 CH 2 S), 2.91 (m, 1H; S-CH (CH 2 g), 3.01 (m, 1H, CH-13), 3 , 71 (m, 1H; CH-3), 4.21 (m, 1H; CH-16), 4.28 (m, 1H; CH-11) and 5.08 (m, 1H, CH-24) ppm Tetramethylsilane was used as internal reference.

Examples 26-28

Following the procedure of Example 25, but replacing isopropylmercaptan with the mercaptans shown in Table XI, the compounds shown in Table XI were prepared.

Table XI: H3 <\ x ^ H3

COOH

UA I

'^ "X ^ V ^ SC ^ CH2 SR

ChJ CHj_J

- "V H3 HO 'ch3

Resulting compounds

Example Mercaptan R Melting point (° C) 26 ethylmercaptan CH CHCH CH 149-152 27 t-butylmercaptan CCHCHH 134 134 134-135 28 cyclohexylmercaptan cyclohexyl amorphous NMR spectrum of the compound of Example 28 (CDCl ^) showed signals at <f = 0.99 (s, 6h), 1.37 (s, 3H), 1.6, and 1.68 (2 bs, 6h), 2.78 (bs, 4H; SCH 2 CH 2 S), 3.07 (m, 1Η) ; CH-13), 3.76 (m, 1H; CH-3), 4.26 (d, 1H; CH-16), 4.35 (m, 1H; CH-11) and 5.12 (m , 1H; CH-24) ppm. Tetramethylsilane was used as internal reference.

4ο 143905

Example 29 l6-Deacetoxy-16B- (2-fluoroethylthio) fusidic acid A. Benzoyloxymethyl ester of 3-O-formyl-l6-deacetoxy-16B- (2-bromoethylthio) fusidic acid l6-Deacetoxy-l6p- (2-hydroxyethyl) (53, ¾ mg) was dissolved in methanol (10 ml) and converted to the sodium salt by titration with 2N sodium hydroxide with phenolphthalein as indicator. After evaporation in vacuo, the amorphous sodium salt was dissolved in dimethylformamide (7.5 ml), added with chloromethyl benzoate (0.16 ml) and the mixture was stirred at room temperature for 48 hours. Water (50 ml) was added and the mixture extracted with ether (100 ml). The organic phase was separated, washed with water (4 x 100 ml), dried and evaporated in vacuo.

. 1

The benzoyloxymethyl ester of 16-deacetoxy-16β- (2-hydroxyethylthio) fusidic acid was isolated as an amorphous product. This was dissolved in dimethylformamide (10 ml) and phenyl-N, N-dimethylformimidate bromide (1.5 s) was added with stirring, after which the solution was allowed to stand at 20 ° C for 24 hours. Water (50 ml) and ether (50 ml) were added and the organic phase was washed with 2N sodium hydroxide (2 x 25 ml) and water (2 x 25 ml), dried and evaporated in vacuo to give the benzoyloxymethyl ester of 3- O-Formyl-16-deacetoxy-16β- (2-bromethylthio) fusidic acid was isolated as an amorphous product.

B. l6-Deacetoxy-l6B- (2-fluoroethylthio) fusidic acid

The substance prepared under A was dissolved in acetonitrile (25 ml), silver fluoride (500 mg) was added and the resulting suspension was stirred at room temperature for 2 hours. Ethyl acetate (50 ml) was added and insoluble material was filtered off. The filtrate was evaporated in vacuo, the residue dissolved in methanol (10 ml) and potassium carbonate (350 mg) added. After stirring for 30 minutes at room temperature, water (100 ml), 4N hydrochloric acid (5 ml) and ether (100 ml) were added and the organic phase was separated.

It was washed twice with water, dried and evaporated to an amorphous product, which was purified by chromatography (ether: acetic acid, 100: 0.5) to give pure 16-deacetoxy-16β- (2-fluoroethylthio) fusidic acid which was crystallized from ether. petroleum ether, mp 157-159 ° C.

Example 30 k in 143905 H-Keto-16-deacetoxy-16P-isopropylthiofusidic acid A. p-Nitrobenzyl ester of 3-O-acetyl-11-keto-16-deacetoxy-6β-isopropylthiofusidic acid

Pyridine chlorochromat (1.07 g) was suspended in methylene chloride (30 ml) with stirring and p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16β-isopropylthiofusidic acid (1.5S) was rapidly added. After stirring for an additional hour, the suspension was diluted with ether (100 ml), the solvent decanted off and the black residue washed twice with ether. Filtration and evaporation of the combined organic extracts gave an oily residue which crystallized from ether-petroleum ether. The colorless crystals were collected, washed with petroleum ether, dried to give 880 mg of the desired product, mp 120-122 ° C.

B. H-Keto-16-deacetoxy-163-isopropyl thiofusidic acid

The above-prepared p-nitrobenzyl ester was dissolved in a mixture of ethanol (20 ml) and 2N aqueous sodium hydroxide (5 ml) and heated to 60 ° C for 3 hours. Then, with stirring, k N hydrochloric acid (3 ml), water (100 ml) and ether (100 ml) were added. The organic phase was separated, washed twice with water (25 ml), dried and evaporated in vacuo. The oily residue was purified by chromatography on silica gel (cyclohexane: ethyl acetate, 7: 3) to give 380 mg of H-keto-16-deacetoxy-16β-isopropylthiofusidic acid, m.p. l67-169 ° C (crystallized from ether-petroleum ether) .

Example 31 11-Keto-16-deacetoxy-16P-isopropylthio-24,25-dihydrofusidic acid Following the procedure of Example 30, but replacing the p-nitrobenzyl ester of 3-O-acetyl-16-deacetoxy-16β-isopropyl-thiofusidic acid with p -nitrobenzyl ester 3-O-acetyl-l6-deacetoxy-16β-isopropylthio-24,25-dihydrofusidic acid was prepared as two-l6-deace toxy-l63-i sopropylthio-24,25-dihydrofusidic acid as colorless crystals, mp 189-191 ° C · 42 1439Ό5

Example 32 3-Keto-16-deacetoxy-163-isopropylthlofusidic acid

To a solution of acetoxymethyl ester of 16-deacetoxy-163-isopropylthiofusidic acid (2.0 g) in 15 ml of dimethyl sulfoxide was added dicyclohexylcarbodiimide (30 g) and orthophosphoric acid (160 mg) and the mixture was stirred at room temperature for 24 hours. A solution of oxalic acid (3 g) in methanol (20 ml) was then added to destroy excess carbodiimide and stirring was continued for 30 minutes. Ethyl acetate (150 ml) was then added and the resulting solution was washed with saturated aqueous sodium bicarbonate (2 x 50 ml) and with water (50 ml). It was dried to give 1.9 g of oil after evaporation. This was dissolved in methanol (4θ ml) and potassium carbonate (1.2 g) was added. After stirring for 1 hour, the methanol was evaporated in vacuo and the residue was added ether (100 ml) and 4N hydrochloric acid (50 ml). The organic phase was washed with water (2 x 50 ml), dried and evaporated in vacuo. The residual oil was purified by chromatography on silica gel (cyclohexane: ethyl acetate, 7: 3) to give 3-keto-16-deacetoxy-163-isopropylthiofusidic acid as colorless crystals, mp 200-203 ° C.

Example 33 16-Deacetoxy-16β-isopropylsulfinylfusidic acid

A solution of sodium metaperiodate (6 g) in 500 ml of water was added to a solution of 16-deacetoxy-163-isopropylthiofusidic acid (10.0 g) in a mixture of methanol (200 ml) and 2 N sodium hydroxide (10 ml). After standing for 1.5 hours, the solution was acidified with 4 N hydrochloric acid (7> 5 ml), which separated a crystalline product. The crystals were filtered off, washed with water (50 ml) and dried and 10 g of the desired product were isolated at mp 158-159 ° C. The crystals were transferred in another crystal modification by treatment with boiling ethyl acetate (400 ml). After cooling to 0 ° C, the product was filtered off, washed with ether (50 ml) and dried. 9 »θ4 g of pure 16-deacetoxy-163-isopropylsulfinylfusidic acid, m.p. 179-118 ° C, were obtained.

Examples 34-39

Following the procedure of Example 33 but replacing 16-deacetoxy-163-isopropylthiofusidic acid with the 163-thioethers shown in Table XII, the sulfoxides shown in Table XII were prepared.

43 143905

HaC CHo

Table XII: J

LxCOOH

c ^ l ch3_] 5 {J ch3 HO '' i CH ^

Resulting compounds

Example Thioether of l6-deacet-R Melting point oxy-fusidic acid (C) 34 δδ-methylthioether CH ^ 151-156 35 16β-ethylthioether CH2CH3 158-162.5 36 Ιββ-t-butylthioether θ (θΗ ^) 3 164-167 37 16β - (2-hydroxyethyl- CH_CH_OH 163-168 thioether 38 ΐ6β- (2'-azidoethylthio) - CH2CH2N 144-147 ether 39 δδβ-phenylthioether CgHH amorphous

Example 40 16-Deacetoxy-163-isopropylsulfinyl-24,25-dihydrofusidic acid Following the procedure of Example 33, but replacing 16-deacetoxy-163-isopropylthiofusidic acid with 16-dacetoxy-163-isopropylthio-24,25-dihydrofusidic acid -deacetoxy-16β-isopropylsulfinyl-24,25-dihydrofusidic acid as colorless crystals, mp 184-186 ° C.

Example 41 16-Deacetoxy-3-keto-163-isopropylsulfinylfusidic acid

Following the procedure of Example 33, but substituting 16-deac an oxy-16 β-isopropyl thio fus idenoic acid with 16-deacetoxy-3-lceto-1,6β-isopropylthiofusidic acid, 16-deacetoxy-3-koto-16β-isopropylsulfinyl fusidic acid was prepared as colorless crystals, m.p. 158-166 ° C.

143905 44

Example kZ

ll-keto-16-deacetoxy-163-isopropylsulfinylfusidinsyre

To a solution of 16-deacetoxy-16β-isopropylsulfinyl-fusidic acid (1.1 g) in 5 ml of pyridine was added acetic anhydride (0.8 ml). After standing for 48 hours at room temperature, 1 ml of water was added to the solution, which after a further 1 hour was diluted with 50 ml of ethyl acetate, washed twice with k N hydrochloric acid and twice with water. The solution was then dried and evaporated to give 94-0 mg of crystalline 3-O-acetyl-16-deacetoxy-16β-isopropylsulfinylfusidic acid, mp 176-178 ° C.

To a suspension of 770 mg of this substance in acetone (100 ml) was added Jones reagent (0.78 ml). After standing for 10 minutes at room temperature, water (100 ml) was added and the resulting solution was concentrated in vacuo to 125 ml to precipitate 3-O-acetyl-11-keto-16-deacetoxy-16β-isopropylsulfinylfusidic acid as colorless crystals which were filtered off. After washing with water and drying, 570 mg of melting point 151-160 ° C was obtained.

400 mg of this product was dissolved in a mixture of ethanol (20 ml) and 2 N aqueous sodium hydroxide (2 ml) and allowed to stand at room temperature for 6 days. Then, 4 N aqueous hydrochloric acid (2 ml) was added with stirring to precipitate the desired product as colorless crystals, which were collected, washed with water (15 ml) and dried. 230 mg were obtained, mp 174-178 ° C. Recrystallization of ethyl acetate gave the pure 11-keto-16-deacet-oxy-16β-isopropylsulfinylfusidic acid, mp 181-183 ° C.

Example 43 3.11-Diketo-16-deacetoxy-16β-isopropylsulfinylfusidic acid

To a solution of 16-deacetoxy-16β-isopropylthiofusidic acid (500 mg) in 100 ml of acetone was added 1.6 ml of Jones reagent. After standing at room temperature for 10 minutes, 100 ml of water was added with stirring. The white precipitate which formed was filtered off, washed with water and dried to give 450 mg of a mixture of 3 * 11-diketo-16-deacetoxy-163-isopropylthiofusidic acid and the desired product.

The crystals were dissolved in hot ether (20 ml) and, upon cooling to 0 ° C, pure 3> H-diketo-16-deacetoxy-16β-isopropylsulfinyl-fusidic acid precipitated. The crystals were filtered off, washed with cold ether and dried to give 60 mg of mp 154-162 ° C.

Example 44 k5 143905 l6-Deacetoxy-l6P-ethoxyphusidic acid

To a suspension of phenacyl ester of 3-O-acetyl-16-deacet-oxy-16α-bromfusidic acid (20.9 µg) in ethanol (300 ml) was added silver carbonate (16.55 g), the mixture was protected from light and was stirred at room temperature for 18 hours. Undissolved material was filtered off and washed with ethanol (2 x 30 ml). To the combined filtrate and wash was added 5N aqueous sodium hydroxide (120 ml) and the mixture was refluxed for 2 hours. After cooling to room temperature, the majority of ethanol was removed in vacuo and to the residue was added ethyl acetate (150 ml) and water (100 ml). With stirring, the mixture was acidified with k N hydrochloric acid. The organic phase was separated and the aqueous phase was again extracted with ethyl acetate (50 ml). The combined organic extracts were washed with water, dried and evaporated in vacuo to an oil which crystallized from diisopropyl ether. The colorless crystals were collected, washed with diisopropyl ether and dried to give 5> 2 g of 16-deacetoxy-16β-ethoxy-fusidic acid, m.p. 169-171 ° C. After working up the mother liquor, 2.20 g of the desired substance were obtained, mp 168-170 ° C. Two recrystallizations of methanol-dlisopropyl ether gave the analytically pure substance, mp 177-17tf ° C.

Examples 45-47 16-Deacetoxy-163 alkyloxy fusidic acids

Following the procedure of Example kk but replacing ethanol with the alcohols shown in Table XIII, the 16-deacetoxy-16β-alkyloxy fusidic acids were prepared in Table XIII.

Table XIII: (next page) hc ch

k ^ COOH

HAY

CH, CH, T

[Λ * J ch3 “3 46 143905 _ Resulting compounds

Example Alcohol R Melting point (C) 45 Methanol CH 3 175-176 46 2,2,2-Trifluoroethanol CH 2 CF 2 202-203 47 Hexanol- (1) CH 2 (CH 2)

Example 48 16-Peacetoxy-160- (2-fluoroethoxy) fusidic acid

Benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16a-bromo-fusidic acid (8.75 S) was dissolved in 2-fluoroethanol (25 ml) and silver carbonate (6.89 g) was added. The mixture was stirred at room temperature and excluded from light for 16 hours. Undissolved material was filtered off, washed twice with ether and the combined filtrate and wash ether were evaporated to dryness in vacuo.

The residual oil containing the crude benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16β- (2-fluoroethoxy) fusidic acid was dissolved in methanol (85 ml), potassium carbonate (3.6 g) was added and the mixture was stirred. stirred at room temperature for 30 minutes. The majority of the solvent was removed in vacuo and to the residue was added water (100 ml) and ether (100 ml). With stirring, the mixture was acidified with 4N hydrochloric acid. The organic layer was separated, the aqueous layer was extracted again with ether (50 ml) and the combined organic phases were washed with water for neutral reaction.

To separate the desired product from the by-product 16-deacetyl-fusidic acid lactone, the ether solution was extracted with 0.5 N sodium hydroxide (3 x 50 ml) and washed with water (3 x 25 ml).

To the combined aqueous phase and wash water was added ether (100 ml) and, with stirring, the mixture was acidified with 4 N hydrochloric acid. The organic layer was separated and the water layer was extracted with ether (50 ml). The combined organic extracts were washed with water for neutral reaction, dried and evaporated in vacuo. The amorphous residue was dissolved in diisopropyl ether (30 ml) and, by scraping, a crystalline product precipitated. After standing in the refrigerator overnight, the crystals were filtered off, washed with diisopropyl ether and dried. There was thus obtained 2.32 g of 16-deacet-oxy-16β- (2-fluoroethoxy) fusidic acid, mp 158-160 ° C.

From the mother liquor, an additional 0.48 g of melting point 155-159 ° C could be recovered. Two recrystallizations of methanol-diisopropyl ether gave the analytically pure product, m.p. 162-163 ° C.

Examples 49-50 47 143905 l6-Deacetoxy-l6P alkyloxy fusidic acids

Following the procedure of Example 48 but replacing 2-fluoroethanol with the alcohols shown in Table XIV, the 16-deacetoxy-16β-alkyloxy-fusidic acids shown in Table XIV were obtained.

H + C CH

Table XIV: 3 X ^ COOH H0. 1 ch3 ch3

Resulting compounds

Example Alcohol R Melting point ° g ^ 49 2-Acetoxyethanol CH2 CH2 OH 179-182 50 1,3-Difluoropropanol- (2) CH (CH2F) 216y-171

Examples 51-60 l6g-Ethers of 16-deacetoxy fusidic acid

Following the procedure of Example 48, but replacing benzoyloxymethyl ester of 3-O-formyl-16-deacetoxy-16a-bromo-fusidic acid with acetoxymethyl ester of 16-deacetoxy-16ctr.bromo-fusidic acid, and replacing 2-fluoroethanol with the alcohols shown. in Table XV, the 16β-ethers of 16-deacetoxy fusidic acid prepared in Table XV were prepared.

Table XV; (next page) h3cc: h3

X ^ COOH HO I

CH, CH T

jjL i y_i H0 '' | ^ Sv ^ “3 48 143905

Resulting compounds

Example Alkobol R Melting point (° C) 51 Isopropanol CH (CH 2) 2 189-190 52 tert-Butanol C (CH 2) ^ 179-180 53 2,2-Dichloroethanol CH 2 CHCl 2 181-182 54 2,2,2-Trichloroethanol CH 2 CCl 2 212-213 55 1,3-Difluoropropanol- (2) CH (CH 2 F) 2 169-171 56 1,3-Diacetoxypropanol- CH 2 CH-OH) CH2 154-156 58 2-Butenol- (1) CH CH = CHCH 128-135 (decomposition) 59 2-Propynol- (1) CH2C = CH 134-136 60 Cyclopentanol cyclopentyl 188-189

Example 61 16-Peacetoxy-168-ethoxy-24,25-dihydrofusidic acid A. Pivaloyloxymethyl ester of 3-O-acetyl-16-epideacetyl-24t25-dihydrofusidic acid

To a solution of 3-O-α-cetyl-16-epideacetyl-24,25-dihydrofusidic acid (31 »12 g) in dimethylformamide (250 ml) was added triethylamine (l, 92 ml) and after 15 minutes stirring chloromethyl pivalate (17.76 ml). After stirring for 20 hours at room temperature, the mixture was diluted with ethyl acetate (750 ml) and washed thoroughly with water (4 x 250 ml, 2 x 50 ml) to remove unreacted starting material and most of dimethylformamide. The organic phase was dried and evaporated in vacuo to give an oily residue. This was dissolved in ether (50 ml), petroleum ether (200 ml) was added and the mixture was stirred for 2 hours. The crystalline precipitate was filtered off, washed with ether: petroleum ether 1: 4.

The combined filtrate plus the wash phases were evaporated in vacuo to give 36 g of crude pivaloyloxymethyl ester of 3-O-acetyl-16-epideacetyl-24,25-dihydrofusidic acid as a foam which did not crystallize.

B. Pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16a-methanesulfonyloxy-24,25-dihydrofusidic acid

To a solution of the crude product prepared above (30 g) in methylene chloride (75 ml) and pyridine (75 ml) was added - Stirring was added dropwise a solution of methanesulfonyl chloride (13.8 ml) in methylene chloride (25 ml) at a temperature of -20 ° C.

After the addition was complete, (about 15 minutes), the mixture was stirred at -15 ° C for 1.5 hours and then left to refrigerate overnight. Ice (about 15 g) was added and after stirring for 0.5 hour the mixture was poured into a mixture of ether (25 ml) and water (100 ml) and shaken vigorously. The organic layer was separated and the aqueous phase was extracted again with ether (100 ml).

The combined organic phases were washed with water, 4N hydrochloric acid (to remove pyridine), saturated aqueous sodium chloride, 0.5 M aqueous sodium bicarbonate, and again with a saturated aqueous sodium chloride. Then it was dried and evaporated in vacuo to a light foam of pivaloyloxymethyl ester of 3-O-acetyl-16-deacet-oxy-16oc-methanesulfonyloxy-24,25-dihydrofusidic acid. The rather unstable product was used in the next step without further purification. It had an IR spectrum: IR (KBr): 1170 and 1365 cm C. Pivalovloxymethyl ester of 3-O-acetyl-16-deacetoxy-168-ethoxy-24,25-dihydrofusidic acid

A solution of the above crude product (2.6 g) in ethanol (25 ml) was stirred at 60-65 ° C for two hours. Water (100 ml) was added and the mixture was extracted with ethyl acetate (2 x 25 ml).

The combined organic phases were washed with water, dried and evaporated in vacuo to a yellowish gum. This was purified by chromatography on silica gel (cyclohexaniethyl acetate 85:15) and Sav 0.72 g of pivaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16β-ethoxy-24,25-dihydrofusidic acid as a colorless foam.

D. l6-Peacetoxy-l68-ethoxy-24,25-dihydrofusidic acid

To a solution of the ΐόβ-ethoxy ester described above in ethanol (10 ml) was added 5N aqueous sodium hydroxide (2 ml) and the mixture was allowed to stand 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, dried and evaporated in vacuo. 0.52 g of an amorphous substance was obtained which crystallized by the addition of ether. The crystals were filtered off, washed with ether and dried to give 0.26 g of 16-deacetoxy-16β-ethoxy-24,25-dihydrofusidic acid, mp 189-191 ° C. Two recrystallizations of ether gave an analytically pure product, mp 192-193 ° C.

Example 62

50 1439OS

λ16-Deacetoxy-16B-methoxy-24.25-dihydrofusidic acid,., ¾. A. Pivaloyloxymethyl ester of 3-Q-acetyl-16-deacetoxy-163-methoxy-24,25-dihydrofusidic acid

Following the procedure of Example 61 A-C but replacing ethanol with methanol, pyaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16β-methoxy-24,25-dihydrofusidic acid was prepared.

B, 16-Deacetoxy-163-methoxy-24,25-dihydrofusidic acid

Following the procedure of Example 6D but replacing the 16β-ethoxy derivative with the δδ-methoxy derivative, 166-deacetoxy-163-methoxy-24,25-dihydrofusidic acid was prepared, mp 152-154 ° C.

Example 6.3 16-Deacetoxy-163-propyloxy fusidic acid A. Piyaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16g-methanesulfonyloxy fusidic acid

A solution of methanesulfonyl chloride (4.6 ml) in methylene chloride (10 ml) was added dropwise at -20 ° C and with stirring to a solution of crude pyaloyloxymethyl ester of 3-O-acetyl-16-. epideacetyl fusidic acid (10 g) in a mixture of methylene chloride (25 ml) and pyridine (25 ml) placed in a three-necked 250 ml flask equipped with a thermometer, separating funnel and drying tube. After the addition was complete, the mixture was stirred at -15 ° C for another 1.5 hours and then refrigerated overnight. Ice (5 g) was added and after stirring for 0.5 h the mixture was poured into water (50 ml) and extracted with ether (2 x 50 ml). The combined organic phases were washed with water, h N hydrochloric acid (to remove pyridine), saturated aqueous sodium chloride, 0.5 M aqueous sodium hydrogen carbonate, and again saturated aqueous sodium chloride.

Then, dried and evaporated in vacuo to give 10.6 g of crude pyaloyloxymethyl ester of 3-O-acetyl-16-deacetoxy-16a-methanesulfonyloxy fusidic acid as a yellow amorphous product.

The unstable product was used in the next step without further purification.

IR (KBr): 1170 and 1355 cm ”1

Following the procedure described above but replacing methanesulfonyl chloride with p-toluenesulfonyl chloride, the corresponding 16 (X-p-toluenesulfonyloxy compound) was prepared.

3i 143905 B. l6-Deacetoxy-l6P-propyloxy fusidic acid

To a solution of crude pivaloyloxyraethyl ester of 3-O-acetyl-16-deacetoxy-16a-methanesulfonyloxy-fusidic acid (1.42 g) in propanol-1 (10 ml) was added triethylamine (0.28 ml) and the mixture was stirred at room temperature for 1 h. 42 hours. After dilution with ethyl acetate (4θ ml), the mixture was washed with water, diluted hydrochloric acid and again with water, dried and evaporated in vacuo to give 1.28 g of an amorphous product. This residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate, 85:15) to give 0.36 g of pivaloyloxyraethyl ester of 3-O-acetyl-16-deacetoxy-16β-propyloxy fusidic acid as a colorless foam.

The ester was hydrolyzed by dissolving in ethanol (5 ml), adding 5 N aqueous sodium hydroxide (1 ml) and refluxing for two hours. After working up as described in Example 61D, the crystalline l6-deacetoxy-l63-propyloxy-fusidic acid was obtained, mp 176-177 ° C.

Examples 64-68 166-Deacetoxy-16β-alkyloxy fusidic acids

Following the procedure of Example 63 but replacing propanol-1 with the alcohols shown in Table XVI, the l6-deacetoxy-163 alkyloxy fusidic acids were prepared in Table XVI.

H + C CH

Table XVI: 3

k / COOH HO I

_J

I I ch3 H0 '"CH3

Resul Tere lead connections

Example Alcohol R Melting point (° c) 64 Butanol- (1) CH2CH2CH2CH3 167-169 65 Isobutanol CH2CH (CH3) 2 189-190 66 2-Methoxyethanol CH2CH20CH3 163-165 67 2-Chloroethanol CH2 CH2 Cl 158-159 68 Benzyl Alcohol CH2C6H5 113-H9 52 143305

Example 69 16-Deacetoxy-168- (2-azidoethoxy) fusidic acid A. Phenacyl ester of 3-O-acetyl-16-deacetoxy-16 | 3- (2-hydroxy-ethyloxy) fusidic acid

To a solution of the phenyl ester of 3-O-acetyl-16-deacetoxy-16a-bromfusidic acid (13 »96 g) in a 1: 1 mixture of ethylene glycol mono and diacetate (80 ml) was added silver carbonate (11, 03 g) The mixture was protected from light and stirred at room temperature for 3 days. Undissolved material was filtered off and washed with ether (2 x 20 ml). After removing the solvent from the combined filtrate and washing ether under reduced pressure, the liquid residue was diluted with methanol (320 ml) and potassium carbonate (5.53 s) added. The mixture was stirred for 30 minutes at room temperature and evaporated in vacuo. The remaining oil was dissolved in a mixture of ether (200 ml) and water (200 ml).

With stirring, the mixture was acidified with dilute hydrochloric acid, the organic phase separated and the aqueous phase extracted with ether (100 ml). The combined organic extracts were washed with water for neutral reaction, dried and evaporated in vacuo.

The yellowish amorphous residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate, 6: 4) to give 5.54 g of the desired product as a colorless amorphous powder which did not crystallize.

B. Phenacylester of 3-O-acetyl-16-deacetoxy-16β- (2-bromoethoxy) fusidic acid

Phenyl ester of 3-O-ηοβΐν1-ΐ6 ^ βησβΐοχν-ΐ6β- (2-hydroxy-ethyloxy) fusidic acid (4.21 g) was dissolved in dimethylformamide (25 ml) and phenyl-N, N-dimethylformimidate bromide (4.6) was added. g), after which the mixture was stirred for 16 hours at room temperature.

After dilution with ether (100 ml), the mixture was washed with water (4 x 25 ml) and the organic phase was dried and evaporated in vacuo. The residual oil was purified by chromatography on silica gel (petroleum ether: ethyl acetate, 85:15) to give 3.16 g of phenacyl ester of 3-O-acetyl-16-deacetoxy-1,66- (2-bromoethoxy) fusidic acid as an amorphous colorless product.

C. Phenacylester of 3-O-acetyl-16-deacetoxy-16 (3- (2-azidoethoxy) -fusidic acid

A solution of the aforementioned phenacylester (1.04 g) and lithium azide (0.34 g) in dimethylformamide (20 ml) was stirred at room temperature for 16 53 143905 hours. The mixture was then diluted with ether (8θ ml), washed with water (4 x 20 ml) and the organic phase dried and evaporated in vacuo to give a foam of the desired substance.

IR (KBr) s 2100 cm -1 - N

D. l6-Peacetoxy-l68 ~ (2-azidoethoxy) fusidic acid

To a solution of the phenyl ester of 3-O-acetyl-16-deacet-oxy-16β- (2-azidoethoxy) fusidic acid (0.95 g) in ethanol (20 ml) was added 5 N sodium hydroxide (2.7 ml) and the mixture was stirred for 18 hours at room temperature. The solvent was removed in vacuo and the oil residue was dissolved in water (40 ml) and extracted with ether (20 ml). The aqueous phase was separated, acidified with dilute hydrochloric acid, and the precipitated oil extracted twice with ether. The combined ether phases were washed with water, dried, and evaporated. There was thus obtained 0.8 g of an amorphous product which crystallized from diisopropyl ether to give 0.4 l of 16-deacetoxy-ΐ6β (2-azidoethoxy) fusidic acid, m.p. 179-182 ° C. Two recrystallizations of the same solvent gave the analytically pure product, mp 184-185 ° C.

Example 70 16-Deacetoxy-16g-ethoxy-24,25-dihydrofusidic acid A. Benzyl ester of 16-deacetoxy-168-ethoxy-fusidic acid

To benzyl ester of l6-deacetoxy-l6a-bromfusidic acid (3.14) in ethanol (25 ml) was added silver carbonate (2.76 g) and, with the exclusion of light, the mixture was stirred at room temperature for 16 hours.

Undissolved material was filtered off, washed with ethanol (2 x 5 ml) and the combined filtrate and washings were evaporated in vacuo. Don amorphous residue was purified by chromatography on silica gel (petroleum ether: ethyl acetate, 6θ: 4θ) to give 1.66 g of the desired product as a colorless foam.

B. 16 Peacetoxy-168-ethoxy-24,25-dihydrofusidic acid

To a solution of benzyl ester of 16-deacetoxy-163-ethoxy-fusidic acid (1.2 g) in ethanol (20 ml) was added 10 $ palladium on carbon catalyst (0.4 g) and the mixture was shaken in a hydrogen atmosphere for 4θ minutes. The catalyst was filtered off, washed with ethanol and the combined filtrate and washings were evaporated in vacuo. The residue was crystallized from ether to give 0.92 g of 16-deacetoxy-16β-ethoxy-24,25-dihydrofusidic acid, mp 191-192 ° C.

Example 71 5b 143905 l6-Deacetoxy-l6i3- (2 * .2%, 2-trifluoroethoxy) -24.2 5-Dilydrofrofidic acid To a solution of l6-deacetoxy-l6p- (2,2,2-trifluoroethoxy) fusidic acid (278 mg) In 96% ethanol (5 ml), X0% palladium was added to calcium carbonate catalyst (50 mg) and the mixture was shaken in a hydrogen atmosphere for 20 minutes. The catalyst was filtered off, washed with 96% ethanol and the combined ethanol phases were evaporated to dryness in vacuo. The residue was crystallized from diisopropyl ether to give 220 mg of the desired product, mp 204-205 ° C. Recrystallization of the same solvent gave the analytically pure product, mp 204-205 ° C.

Example 12 16-Peacetoxy-168- (2-fluoroethoxy) -24.25-dihydrofusidic acid

Following the procedure of Example 71 but substituting 16-deacetoxy-163 ~ (2,2,2-trifluoroethoxy) fusidic acid with 166-deacetoxy-16β- (2-fluoroethoxy) -fusidic acid, 16-deacetoxy-16β- (2) was prepared. -fluoroethoxy) -24,25-dihydrofusidic acid, mp 180-182 ° C.

Example 7 3 11-Keto-16-deacetoxy-168-ethoxyphusidic acid

To a suspension of the phenyl ester of 3-O-acetyl-11-keto-16-deacetoxy-16a-bromofusidic acid (5 57 g) in ethanol (60 ml) was added silver carbonate (4.4 g) and stirred under light protection. the mixture for 18 hours at room temperature. Undissolved material was filtered off and washed with ethanol (2 x 20 ml).

The combined ethanol solutions containing the crude phenacyl ester of 3-O-acetyl-11-keto-16-deacetoxy-163-ethoxyphusidic acid were diluted with ethanol (80 ml), 5N aqueous sodium hydroxide (32 ml) was added and the mixture was stirred for 20 hours. at room temperature. The solvent was removed in vacuo and to the residue was added water (100 ml) and ether (100 ml). With stirring, the mixture was made acidic by the addition of k N hydrochloric acid. The organic phase was separated, the aqueous phase was again extracted with ether (100 ml) and the combined ether phases were washed with water for neutral reaction. The ether phase was then dried and the ether was evaporated in vacuo. The residual oil was purified by chromatography on silica gel (ether: petroleum ether: acetic acid, 50: 50: 0.5), 55 143905 and the yellowish amorphous product was crystallized from diisopropyl ether to give 2.12 g of H-keto-16 l63-ethoxyphusidic acid, m.p. l66-l67 ° C. Recrystallization of ether-diisopropyl ether gave the analytically pure substance, m.p. 167-168 ° C.

Example 74 11-Keto-16-deacetoxy-163- (2-fluoroethoxy) fusidic acid

Following the procedure of Example 73 but replacing ethanol with 2-fluoroethanol, 11-keto-16-deacetoxy-Ι6β- (2-fluoroethoxy) fusidic acid was prepared as a colorless amorphous powder which could be converted to a crystalline sodium salt (see Example 86 ).

Example 75 3,11-Diketo-16-deacetoxy-163-ethoxyphusidic acid

To a solution of l6-deacetoxy-l63-ethoxyphusidic acid (10.24 g) in acetone (200 ml) was added Jones reagent (l2.0 ml) dropwise at 15 ° C. After the addition was complete, the cooling bath was removed and the mixture was stirred for 30 minutes at room temperature. To the mixture was added ether (100 ml) and water (200 ml) and stirring was continued for 13 minutes. The organic layer was separated, the aqueous phase was again extracted with ether (100 ml) and the combined organic extracts were washed with water for neutral reaction and dried. The ether phase was concentrated to ca. 100 ml, whereby a colorless product crystallized. After standing in the refrigerator overnight, the crystals were collected to give 7.02 g of 3, H-diketo-16-deacetoxy-163-ethoxyphusidic acid, m.p. 185-87 ° C. An additional 0.84 g was crystallized from the mother liquor. Recrystallization of methylene chloride diisopropyl ether gave the analytically pure product, mp 187-188 ° C.

Example 76 3-Keto-16-deacetoxy-163-ethoxyphusidic acid

A mixture of 3, H-diketo-16-deacetoxy-163-ethoxy-fusidic acid (6.0.5 g), 2-ethyl-2-methyl-dioxolane (-1,3) (60 ml) and p-toluenesulfonic acid (0.24 g) is heated under reflux on an electric hot plate for 40 minutes. After cooling to room temperature, ether (200 ml) and pyridine (0.5 ml) were added and the mixture was washed with water (4 x 50 ml). The organic phase was dried and evaporated in vacuo to give a gum of the crude 3-ethylene ketal of 3, 11-diketo acid which did not crystallize.

This was dissolved with stirring in ethanol (140 ml) and at 5 ° C was added portionwise solid sodium borohydride (2 g). Then, the cooling bath was removed and the mixture was stirred for 45 minutes at room temperature. Neutralized with acetic acid, water (420 ml) was added and the precipitated oil was extracted with ether (2 x 150 ml). The combined extracts were washed with water (4 x 25 ml), dried and evaporated in vacuo. The oil residue crystallized from ether-diisopropyl ether to give 3.12 g of the 3-ethylene ketal of 3-keto-16-deacetoxy-16β-ethoxy-fusidic acid, mp 166-169 ° C. Concentration of the mother liquor gave 2.04 g of the same melting point. Two recrystallizations of methylene chloride diisopropyl ether gave the analytically pure product, mp 171-172 ° C.

A solution of the above substance (3 »98 g) in methanol (4θ ml) was acidified with 2N hydrochloric acid (2 ml) and refluxed for 20 minutes on a steam bath. After cooling, water (160 ml) was added and the precipitated oil was extracted with ether (2 x 100 ml).

The organic extracts were washed with water for neutral reaction, dried, and evaporated in vacuo. The amorphous residue was crystallized from ether to give 2.94 g of 3-keto-16-deacetoxy-16β-ethoxy-fusidic acid, mp 175-177 ° C. Recrystallization of the same solvent gave mp 177-179 °.

Example 77 β-Diethylaminoethyl ester of 16-deacetoxy-163-isopropylsulfinyl-fusidic acid

To a solution of the sodium salt of 16-deacetoxy-16β-iso-propylsulfinylfusidic acid (320 mg) in 2 ml of dimethylformamide was added β- (diethylamino) ethyl chloride (θ, θ8 ml). After standing at room temperature for 5 hours, the desired product crystallized.

Water (5 ml) was added and the product was filtered, washed and dried to isolate 310 mg of the desired product, mp 156-158 ° C.

Example 78

Acetoxymethyl ester of l6-deacetoxy-1,68-isopropylsulfinylfusidic acid To a solution of the sodium salt of l6-deacetoxy-l63-isopropylsulfinylfusidic acid (320 mg) in 2 ml of 57 143905 dime thylf orm amide was added chlorine thylacetate (0.0 ml). After standing for 24 hours at room temperature, ethanol (5 µl) and water (5 ml) were added to precipitate the desired product as colorless crystals which were filtered off, washed and dried.

290 mg of melting point 151-153 ° C were obtained. Recrystallization of ethyl acetate-petroleum ether gave a melting point of 1 ° C-1 ° C.

Example 79

Acetoxymethyl ester of 16-deacetoxy-16B-isopropylthiofusidic acid Following the procedure of Example 78, but replacing 16-deacetoxy-16β-isopropylsulfinylfusidic acid with 16-deacetoxy-l63 C.

Examples 80-87

Sodium salts of 16B ethers, 16B thioethers and 16β-alkylsulfinyl compounds of 16-deacetoxyphusidic acid and its 3- and 11-keto derivatives

Crystalline sodium salts of the compounds described in Examples 1, 33j 44, 46, 48, 73, 74 and 76 were obtained by the following procedure: a solution of the acid in methanol was titrated with 2N methanolic sodium hydroxide with phenolphthalein as indicator. After evaporation to dryness in vacuo, the residue was taken up in acetone (about 100 ml). The solution was concentrated to ca. half the volume, and upon scraping crystallized the desired sodium salt. After 2 hours at room temperature, the crystals were collected, washed with acetone and dried to give the pure sodium salt of the desired compound. The sodium salts are shown in Table XVII. Microanalysis, IR spectrum and NMR data for these substances are consistent with their structure. The sodium salts all melt during decomposition.

Where a value is given in brackets in the table, this indicates the approximate temperature at which the decomposition begins.

Table XVII; H ^ C CH ^ 58 143905 l ^ AJOONa "2vJL-» 3

CH. CH | IN

jcdV

'ch3 2_ 2 o acid described Melting point

Example R R A RJ in Example ° C

80 H, cc-OH Η, α-ΟΗ S ChXch ^ J ^ Ϊ 239-245 81 Η, α-ΟΗ Η, α-ΟΗ so ch (ch3) 2 33 175-180 82 Η, α-ΟΗ Η, α -ΟΗ 0 CH ^ H ^ 44> 300 (ca. 250) 83 Η, α-ΟΗ Η, α-ΟΗ o CH2CF3 k6> 280 (ca.24o) 8b Η, α-ΟΗ Η, α-ΟΗ O CHgCHgF 48 199-206 85 Η, α-ΟΗ o CH2CH3 73> 300 (ca.250) 86 Η, α-ΟΗ O CH2CH2F 74 189-203 87 o Η, α-ΟΗ O CH2CH3 76> 300 (ca.260)

Example 88

Kalitun salt of l6-deacetoxy-168- (2-hydroxyethoxy) fusidic acid

A solution of 16-deacetoxy-16β- (2-hydroxyethoxy) fusidic acid (2.64 g, as hemihydrate) in methanol (10 ml) was titrated against phenolphthalein with 2N methanolic potassium hydroxide. After evaporation to dryness in vacuo, the amorphous residue was dissolved in methanol (2.5 ml). Acetone (60 ml) was added and the mixture concentrated to ca. 15 ml at reduced pressure. Colorless crystals precipitated by scraping, filtered off, washed with acetone and dried. 2.32 g of the desired substance were obtained, mp 243-248 ° (with decomposition).

Claims (1)

3 Y? 3 'C00R'> In vb f \ [CH, Q1i # yv ^ 30 H3 1 "1 H 2 in which Q 1 is for Q or a group, R is an alkanoyl, 2 aralkanoyl or an aroyl group, Q is as as defined above, Y 3 represents chloro, bromo or iodo, and R 2 represents a straight or branched alkyl group having from 1 to 6 carbon atoms, an aralkyl, alkanoyl or aroylmethyl group, an alkanoyl or aroyl oxy -alkyl group, an alkyloxymethyl or cyanomethyl group, is reacted with a compound of formula VII: R -AH in which R 1 is as defined above and A represents oxygen or sulfur to form a compound of formula VIII: H3C cn3 J: *. Y COOR-3 Q2 jT A-R1 c ^ 4_f VI11 cb3 12 1 3 in which QQ, R and R are as defined above and A is oxygen or sulfur, which compound is then, if desired, subjected to a hydrolysis to a free acid of formula I and a compound of formula VIII or I in which A stands for sulfur if desired is oxidized to the corresponding compound where A represents a sulfinyl group and wherein, if desired, a free acid is converted to the corresponding salt or a light hydrolyzable ester wherein R has the above meaning except H, or where compounds in which Q1 2 H and / or Q represent, if desired is oxidized to a compound r 2 of formula I wherein Q and / or Q are oxygen, or wherein a compound in which Q and Q is oxygen is, if desired, reduced to a 1 2 compound of formula I wherein Q or Q is or wherein a compound of formula I in which there is a double bond between C-2k and C-25 is, if desired, hydrogenated to a compound having a single bond between C-24 and C-25, or b) a compound of formula IV HC CH3 J: ^ [COOR3 Q2 J VV A ^ OH IV Crf3T CHJ, Χυ1 "· CH3 - 13905 62 12 3 1 which Q ·, Q and R are as defined above and the OH group at C, ς is α-oriented, reacted with a phosphine and a compound: R SSR to give a compound of formula VXXI (as shown above), in which an A stands for sulfur and R 1 is an aryl group, while QQ and R are as defined above, which compound is then converted to a compound of formula I, as described above under a), or c) a compound of Formula IV HC CH 2 Cl 2 Y I COOR 3 "T _ / OH IV i \ I CH, ch 3 in which the hydroxyl group at C 1 is α-oriented and 1 2 is as defined above, but different from j group, is reacted with HU- "a reactive derivative of an alkylsulfonyl or an arylsulfonyl acid, thereby forming a compound of formula IX: HV" 3: zy f 3 C0 ° R 2 1 5 IX ^^ rV0R CBsl _J δΗ3 2 3 in which Q, R and the dotted line between C ^ and are as H * - defined above, Q 'stands for oxygen or the group Ώ2 »^> where 2 5. u- R is as defined above and R stands for an alkylsulfonyl or arylsulfonyl group, and the compound formed is then reacted with a compound R 1 -AH wherein R is as defined above and A is oxygen or sulfur to form an compound of formula