DE1937908A1 - Prostaglandin derivatives and processes for their preparation - Google Patents

Description

LAWYER f

DR. JUR. DIPL-CHEM. WALTER BEIL ALFRED HOEPPENER

DR. JUR. DIPL-CHEM. H.-J. WOLFF DR. JUR. HANS CHR. BEIL _v

623 FRANKFURT AM MAIN-HOCHST 'ADEi-ONSIRASSE 58

1937908 June 4, 1969

Our number 15

The Upjohn Company Kalamazoo, Mich., V.St.A.

ProstaKland derivatives and processes for their production

The present invention relates to new compounds, processes for their preparation and those occurring in these processes Intermediates. In particular, the present invention relates to new analogs of prostaglandin E.,

₁ ) ,. Prostaglandin E _1, prostaglandin (PGA ₁₎ (PGP ^ and FGF., _Ss), new processes for the production of PG-E _1, P

₁ I ₃ , PGA ₁ and their new analogs, as well as new chemical intermediates that are of value in these processes.

has the following structure: O

COOH

H . XlH has the following structure:

■ I

COOH

HO-

00982.7 / 2053 "

II

_ß has the following structure:

COOH

IH HO '

PG-A. has the following structure:

COOH

For the stereochemistry of PGE ₁ , PGF ₁ ^, PGF _1ß and see "Nature" page 212, volume 38 (1966).

In the formulas I, II, III and IY and the formulas below, the dashed lines on the cyelopentane ring show substituents in the c. -Configuration on, ie substituents below der.Ebene of the cyclopentane ring, thick lines on the Cyelopentanring indicate substituents in the ß-configuration, ie. Substituents above the level of the cyclopentane ringa.

PGE _1, PGF _1ß, PGF _1ß and PGA ₁ are derivatives of prostanoic acid which has the following structure with the indicated numbering; -;

COOH

Prostanoic acid is systematically referred to as ⁼ 7 * - £ (2i3-Oetyl) -cyclopent-1 ^ -yl7-heptanoic acid.

Similar compounds to the compound of Formula V, each but with a terminal carboxyl group on the side chain in the ß-configuration on the cyelopentane ring,

are referred to as 8-iso-prostanoic acids and have the following formula:

COOH

VI

Iso-prostanoic acid is systematically referred to as 7- / L (2ß- Cc "ryl) -c \ alo-pent-1ß-yl ~ -heptanoic acid. ■ ■ -.

Prostaglandin 3 and its analogues, which are produced according to the new invention Procedure obtained v / ground have the following formula; . ". ■ ■

- ^C n ^F or> ~ ^COUR 1 '·

\ ^ - V ■ " ^R 4 VII HO- ':, C -CC .OH H" ^ C _v

. R3 -R ₂ - ■

in dor R ^ hydrogen, an alkyl group with. ¹ to 3 carbon atcnen including, a cycloalkyl group having 1 to 10 carbon atoms, inclusive, an aralkyl group having from 7 to 1.2 carbon atoms, inclusive, phenyl, a through ll: io ~> chlorine atoms or an alkyl group ir.it 1 to 4 carbon at omen including substituted phenyl or an ethyl group substituted in the β-position by 3 chlorine, 2 or 3 bromine or 1, 2 or 3 iodine atoms; R-, hydrogen or an alkyl group having 1 to 5 carbon atoms inclusive, denoted by C to. 3 Fluerateme can be substituted; R_ and R-. . Hydrogen or alkyl groups having 1 to 4 carbon atoms inclusive, meaning; C Ho "denotes an alkylene group with V-'to 8 carbon atoms, inclusive, which is substituted by O to 2 fluoro-rat: re; and in which the bond of the C. Ho _{n group} on the ring in /. - or ß-configuration; the formula also includes the pharmaceutically usable salts when R. is hydrogen.

009827/20 53

■ - * SAD

Prostaglandin P and its according to the new invention Analogs obtained by the following procedure

Formula shown:
ΗΟ

II / \ VIII

ft- ·? Ro

in which R _1, R _2, R ,, R ₄ and C _n H _2n is as defined above for Formula VII have and- · - the binding of the hydroxy and ~ ^c _n ^H 2n ^C0 ^ ^R 1 ~ ^Gru PP ^{c ycle on the} ring means inot- or ß ~ position; where the formula also includes the pharmacologically useful salts when R ₁ is hydrogen. Formula VIII includes compounds in which the hydroxyl and -CH ₂ -COOR groups can be iricr, '> - »<* ·> ß-, ß, p> or ß, ß-configuration.

Prostaglandin A and its according to the new invention Analogs obtained by the following procedure Formula shown:.

^G n ^H 2n- ^C00R 1

; ^H ' / ^ IX

R ₃ R ₂ ......., ■

in which R ₁ , R ₂ , R ^, R. and Op 'have the meanings given above for formula VII and - the bond of the -G _n H _2n -COOR _{1 group} on the ring in "·· - or ß- The formula also includes the pharmacologically usable salts when R ₁ is hydrogen.

The formulas VII, VIII and IX also include the individual isomers in which the configuration of the hydroxy side chain has an R or S configuration, as well as the racemic form (dl) and the individual optically active enantiomers (d and 1). QQ Qg 2 7/2 5 O 3

»" ■■ -

Formula VII represents PGE ₁ when R., R, and R. each denote hydrogen ", Rg is a pentyl group, CH _2n denotes methylene," C _n Hp _n -COOR ₁ in (^ -configuration to the cyclopentane ring and the hydroxy side chain in S configuration is available.

Formula YIII represents PGF ₁ ^ when R ₁ , R, and R, each _{denote hydrogen, R 2 is} a pentyl group, CH _9n denotes hexamethylene, the hydroxy and -C Hp -COOR ₁ group both form the ring in Cv- -S-position and the hydroxy side chain is in S-configuration.

Formula YIII represents PGF _{1 ß} , when R ₁ , R, and R are each hydrogen, Rp. Is a pentyl group, CH _? Hexamethylene means that the hydroxyl group is in the ß-configuration, -CH ₂ -COOR "is in the. <X -configuration to the ring and the hydroxyl side chain is in the S configuration.

Formula IX represents PGA ₁ when R _1,. R, and R. each denote hydrogen, Rp is a pentyl group, C Hp is hexamethylene, the -CH ₂ -COOR ₁ group to the ring is in the • configuration and the hydroxy side chain is in the S configuration,.

All 'compounds which come under the formulas VII, VIII and IX

show the -CH ^ R.CRgR ^ OH side chain in ß-confi-

guration to the ring with a tra, ns C = C bond as in the formulas shown above.

Examples of the alkyl group having 1 to 4 carbon atoms for the compounds of formulas VII, VIII and. IX are methyl, ethyl, propyl, butyl groups and their isomeric forms. Examples of alkyl groups having 1 to 8 carbon atoms inclusive are the groups indicated above, as well as pentyl, hexyl, heptyl, octyl groups and their isomeric forms. Examples of cycloalkyl groups with 3 to . "* 009827/2053

V 6 ■ - - ". '■. ■■■. ■". ■' ■ .'- '.. ■ -

10 carbon atoms including, including alkyl- substituted cycloalkyl radicals are cyclopropyl, ' 2-methylcyclopropyl-, 2,2-dimethylcyclopropyl-, 2,3-diethylcyclopropyl, ^ -Butylcyclopropyl-, cyclobutyl-, · 2-methylcyclobutyl-, 3-propylcyclobutyl-, 2,3,4-triethylcyclobutyl-, Cyclopentyl, 2,2-dimethylcyclopentyl, 3-pentylcyclopentyl, 3-tert-butylcyclopentyl. ~, Cyclohexyl, 4-tert-butylcyclohexyl, 3-isopropylcyclohexyl, 2,2-dimethylcyclohexyl, Cycloheptyl, cyclooctyl, cyclononyl and cyclodecyl groups.

Examples of aralkyl groups with 7 to 12 carbon atoms including are benzyl, phenethyl, 1-phenylethyl, 2-phenylpropyl, 4-phenylbuty1, 3-phenylbuty 1-, 2- (1 -ITaph- ⁼ ethylethyl) and 1- (2-iaphthylmethyl) groups. Examples of phenyl groups substituted with 1 to 3 chlorine atoms or an alkyl group with 1 to 4 carbon atoms including substituted phenyl groups are p-chlorophenyl, m-chlorophenyl, o-chlorophenyl, 2,4-dichlorophenyl, 2,4,6 -Trichlorophenyl-, p-tolyl-, ni-tolyl-, o-tolyl-, p-ethylphenyl-, p-tert-butylphenyl-, 2,5-dimethylphenyl-, 4-chloro-2-methylphenyl - and 2,4-di- 'chloro-3-methylphenyl groups ,,

Examples of alkylene groups having 1 to 8 carbon atoms; including are methylene, ethylene, trimethylene, Tetramethylene, pentamethylene, hexamethylene, heptamethylene, Octaliethylene residues and their isomeric branched-chain forms,

Examples of alky! Groups having 1 to 8 carbon atoms einsehließlich which may be substituted with 1 to 3 fluorine atoms are _i, are the 2-Fluoräthyl-, 2-fluorobutyl, 5-fluorobutyl, 4-Pluorbutyl-, S-fluoropentyl, 4 -Fluoro-4-methylpentyl, 3-fluoroisoheptyl, 8-fluorooctyl, 3,4-difluorobutyl, 4,4-uifluoropentyl, 5,5-difluoropentyl and 5,5,5-trifluoropentyl.

00982772053

Examples of alkylene groups substituted by 1 or 2 fluorine atoms with 1 to 8 carbon atoms are groups with the following formulas:

-CH ₂ CHF-, -CH ₂ CF ₂ -, -CH ₂ CH ₂ CHFCH ₂ -, ₂₂

CH3
-CH ₂ CHCH ₂ CHF-, -

-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ Ch ₂ CF ₂ -.

-, and

_{1 11 113} and PGA and their esters and pharmacologically acceptable salts are extremely effective in achieving various biological reactions. For this reason these compounds are valuable for pharmacological purposes; see, for example, Bergstrom et al., Pharmacol. Rev., Vol. 20, p. 1 (1-968), as well as reference points indicated there. Some of these biological reactions are the systemic lowering of the arterial blood pressure in the case of PGE ₁ , PGF ₁ o and PGA ₁ , measured for example in anesthetized (pentobarbital sodium), pentolonium treated rats with indwelling cannulas in the aorta and right ventricle, furthermore an antihypertensive effect, which in the same way for FGF. was determined; Stimulation of the smooth muscles, as demonstrated, for example, by tests on ileum muscles. strips of guinea pigs, rabbits duodenum or jerboa colon were found; Potentiation of other smooth muscle stimulants; antilipolytic effect, as demonstrated by the antagonism of Bpinephrine-induced mobilization of free fatty acids or inhibition of the spontaneous release of glycerol from isolated fat pads of rats; the inhibition of gastric secretion in the case of PGE ₁ and PGA ₁ , demonstrated in "dogs whose secretion was stimulated by food or hemamine infusion; effects on the central nervous system; reduced adhesion of blood platelets, demonstrated by the adhesion of blood platelets to glass, as well as the HemmxmgQfe # gBl ^ tBA ^ tjfe <sjhen aggregation and thromb

evil formation caused by various physical irritants, such as arterial injury, as well as various biochemical mordants such as ADP, ATP, serotonin, thrombin and collagen.

Because of these biological reactions, these known prostaglandins are valuable in the investigation, contraception, Control or amelioration of a wide variety of diseases and undesirable physiological conditions Birds and mammals, including humans, domestic animals, Small animals, zoological test animals and laboratory animals, e.g. mouseless, rats, rabbits and monkeys,

For example, these compounds are, in particular in mammals, including humans, of value as nasal anti-congestion agents. Be purely for this purpose the compounds in a dose of about. 10, '- g to about 10 mg. pro-al a pharmaceutically usable liquid Carrier or used as an aerosol spray for topical application.

and PGA ₁ are useful in mammals, humans and certain farm animals, such as dogs and , already present; ulcers accelerating. For this purpose, the compounds are injected or infused intravenously, subcutaneously or intramuscularly, with an infusion dose of about G.1 / <g to about 50 / <g per kg of body weight per minute and for an injection or infusion the total daily dose of about 0 , Is 1 to about 20 mg per kg of body weight, and the exact dosage depends on the age, weight and condition of the patient or animal, as well as on the frequency and type of administration.

009827/20 53. ^ «MuL

, PGA ₁ , PGF _{1r /} and PGF _{1 ß} are valuable for preventing platelet aggregation, reducing platelet adhesion and removing thrombi or preventing thrombosis in mammals, including humans, rabbits and rats. For example, these compounds are valuable in the treatment and prevention of myocardial infarction, for the treatment and prevention of post-operative thrombosis, for promoting the patency of vascular transplants after the operation, for the treatment of, for example, atherosclerosis, arteriosclerosis, blood clotting symptoms due to lipemia, and other clinical conditions for whom the cause of the disease is linked to a disturbance in the fat balance or an excessive occurrence of fat in the blood. For these purposes, these compounds are administered systemically, for example intravenously, subcutaneously, intramuscularly and in the form of sterile implants with a longer duration of action. In order to achieve a quick response, for example in emergencies, intravenous administration is preferred. Dosages in the range from about 0.004 to about 20 mg per kg of body weight per day are used, the exact dose depending on the age, weight and condition of the patient or animal, as well as on the frequency and type of administration.

, PGA ₁ , PGF. _{) r>} , and PGF _{1 ß} are particularly valuable as additives to blood, blood products, blood substitutes and other liquids that are used in the artificial circulation and perfusion of isolated body parts, for example limbs and internal organs, which take place outside the body, regardless of whether they are connected to the body belonging to it, separated from it and preserved or prepared for a transplant or connected to a new body. During these circulations and perfusions, clumped platelets tend to block the blood vessels and parts of the circulatory system.

% 009827/2053

clogging systems. This clogging is prevented by the presence of the compounds according to the invention. For this purpose, the compound is gradually, in a single quantity or multiple Kengenanteilen to the circulating blood, separated to the blood of the donor animal, to the perfused body portion, the ocer connected to the body of it is _y to the receiver, all to two or Concerned at a fixed total dose of about 0.001 to 10 mg per liter of circulating fluid. It is particularly useful for the compounds to be used in laboratory laboratories, including white cats, dogs, rabbits, monkeys, and rats, for these purposes in developing new methods and procedures for green and limb transplants. '..'-'-. "', - '...:.

TGE. is particularly effective for stimulating smooth muscles and is also very active in potentiating other known stimulators for smooth muscles, for example. . blow stimulating agent, for example O ^ ytocin, and various ergot alkaloids, "including derivatives thereof and analogs For this reason, PGL, ana parts of this or in -combination with these known Otimulatoren smooth Iiuskulatur in lower air;., precious the usual amounts to curb or for example atonicches uterine bleeding after abortion or birth to prevent, to facilitate the ejector ^ placenta _s and for treatment during dos, iOehenbetts, for these purposes is PGE ₁ by intravenous infusion immediately after abortion or birth in a dose of about 0 , 01 - to about 5-Q / 4.g _{Prö;;; kg;} body weight per minute until reaching the gevränschten effect administered Consecutive doses are intravenÖFC, subcutaneous or intramuscular injection or infusion during puerperium in an amount of 0th , 01 to 2 rag per kg of body weight administered per day, the exact dosage depending on the age, the weight and the to the state of the patient or the animal.

BAD ORlQlNAE: # 0982772053

PGE ₁ , TGA and PGP _{1 r} are useful as hypotensives for reducing blood pressure in mammals, including humans. To this end, the compounds are administered by intravenous infusion in an amount of from about 0.01 to about 50 "g per kg of body weight per minute, or in a single dose or multiple doses of about ? .5 to 500.cg per kg of body weight per day total administered.

.. _, And PGF, are valuable instead of oxytocin ί, ιιτ introduction of the V / before activity at deceptive animals, such as cows, sheep ei ^5, and pigs, and in pregnant women, and indeed r.uii; Birth η time or short ?: before, or in pregnant animals. those of the fetus died about 20 weeks before the birth date. ? ÜR this purpose the connection by i: ITRA \ ^r enöse infusion at a Itesis from 0.01 to bq '' g pi'o kg '.'crpergG-weight per minute until or almost until the end: the level of sweiten Childbirth, that is, the expulsion of the fetus, is administered. i) These connections are also especially intestinal when the baby is born. urr. one or nehroro weeks is exceeded and 'lie natural \. ^r eh2rt have not started, or if 12 :: 60 hour he he; after urin "BIa-. sensprung have passed, ohii ^ -IaL which of course; 'vcL ntäti ^ ity has set in.

As mentioned above, IGE _{1 is} a virksane-r antagonist for the kittels adrenaline inausiex-tc glycation of free fatty acids. From this cir. The reason is this 7erb: .n-iu: ig valuable in the experimental medicine scv / ohl for ir vitro and in vivo tests in mammals, including humans , Including anomalous L: \ - poid mobiliBiorung · ΐϊηά hbhtr fi-cier photoic acid levelX, - / ie • for example Diacetes mellitus, J-vascular diseases nrv.'ie thyroid hypertrophy, see below recognize, see below, the

009827/20 53 « _To

BATH

To alleviate and cure symptoms. '■■'. "..

The other compounds represented by Formulas YII, YIII and IX, with the exception of PGE. , PGF _{1 r} , Τ? (ΪΈ * ~ and PG-A, also cause one or more of the above listed "biological reactions. The natural prostaglandins PGE ₁ , PGF ₁ ^ and PGA ₁ as well as the PGE ₁ reduction product PGF ₁ "However, it causes numerous reactions even in low doses, PG-J ¹ L, for example, brings about a reduction in blood pressure and stimulation of the smooth muscles and shows - at the same time anti-lip ο lyrical effect. In contrast to this, the compounds that differ from these natural prostaglandins are Formulas .YlI ^: -, .YIII and; IX, much more specific for. The induction of prostaglandin-like biological reactions. Each of these "compounds" is used instead of one of the natural prostaglandins for at least one or more of the pharmacological agents given for the latter For other purposes, they are more valuable than natural prostaglandins, since there is never any difference between them have a different and narrower range of effects and, as a result, more specific in their effects ^: are, with smaller and less unrestricted ebera effects than with natural prostaglandins, in addition, some of these artificial prostaglandins have a greater degree of effectiveness; Eliciting one or more of the above-described 1aolc £, i '; chfcn reactions as - the corresponding natural compound within .fit ::, "ahrnenc of the same ■'

YII, YIII or IX. . . ... .

2ebeispielsv; ei3e in PGE _{1 is} the - (CH ^ j ^ -CCüE-G group in. · Configuration pi \ ä.en CyclopentanriiiK; 4er rorrnel YIX gebuiir-; ■ den. "The corresponding compound, in isr this Ώ- ruppe in ß-Xonfiguraticn stent, i.e. S-ico-POli .., has: only ^ a small part of the effect of PGE ^. hiririnhtlioli of the stimulation of the smooth muscles and the lowering of the blood pressure, has

0098 27/20 53 8ADORIQiNAL

however, a substantial inhibitory effect on platelet clotting and the mobilization of free fatty acids caused by adrenaline.

In the case of PGE ₁ , the configuration of the hydroxy side chain is also an S configuration. If the hydroxy side chain is in the R configuration, ie 15 (R) -PGE., The compound shows only a small part of the effectiveness of PGE _{1 in} terms of lowering blood pressure and antagonism against the mobilization of free fatty acids caused by adrenaline, during the stimulating effect on the smooth muscles is essentially retained.

By substituting PGE. fluorine in the 3-position (see formula Y) gives a compound with about the same effect on the smooth muscles as PGE ₁ , but with less than a third of the effect of PGE _{1 in} terms of lowering blood pressure.

By extending the alkyl chain of PGE ₁ (R ₂ _in formula VII) from Pen.tyl to hexyl, a compound is obtained with the mohr than fourfold effect of PGE ₁ with regard to the inhibition of platelet aggregation caused by ADP, et v / a. 25 r stronger effect in terms of stimulating flor smooth muscles, but with less effect than PGE _{1 in} terms of lowering blood pressure.

Although all of the compounds represented by Formulas VII, VIII and IX are useful for one or more of the _{purposes set out above for PGE 1} , PGF ₁ ,, PGF _{1 β} and PGA ₁ , some of these compounds have particular value in that they are of a substantially longer duration _{Have a longer} duration of action than the other compounds of the general formulas, including PGE ₁ , PGF 1y, PGF _1ß and PGA ₁ , and since they are more oral, siibllngual, intra, vaginal or rectal than,

009827/2053

- 14 - '■.' ■ ', ■■■ ■

as usual, by intravenous, intramuscular or subcutaneous Injection or infusion can be administered as above in the use of the known prostaglandins and the other, given by the formulas YII, YIIi and ΐχ Compounds, which has been explained, are advantageous in that these properties are retained because they are more uniform These compounds mirror in the body with fewer, shorter ones or lower dosages and allow administration by the patient himself.

These special compounds have the following formulas; '"..

Ö09827 / 2053 BADORlCaINAL-

HO "'

(CH ₂ ) nr Z-COOR1

CHOH- (CH ₂ ) p _{-CH 3}

HO ''

(CH ₂ ) _m -Z-COOR _I3

C = C

CHCH- (CH ₂ ) _p -CH _; Xl

c = c:

CHOH- (CH ₂ ) p _{-CH 3}

HO ''

C = C

CHCH- (CH ₂ ) _a -Y XIIi

HO- '

C = C

CHOH- (CH ₂ LY XIV

) n "C00R ₁₃

CHOH- (CH ₂ ) _a -Y XV

(CH ₂ LZ-COOR ₁

CHOH- (CH ₂ ) _a -Y XVI

009827/2053

. · '■■■■ ■'.: ■ / ' - 16 - ■' ■ '■''.■;' ■ "■; /, ■■ '.' ■■ '>.. -; HO

, 'C = C

^H0 'H ^ "^ CHOH- (CH ₂ ) aY XVjI

(CH ₂ ) _m -Z-COOR ₁₃

G = CiT

J ₆ -COOR ₁₃

Hf) '^ CCv,

H ^ - CHOH- (CH ₂ ) _b -CH ₃ XIX.

HO-.

J ₆ -COOR ₁₃

μη * '. C = C ν

H ^ ^ CHOH- (CH ₂ ) _b -CH ₃ XX

(CH ₂ ) a-COOR ₁ a

C = f *

^ CHOH- (CH ₂ Jb-CH ₃

Je-COOR ₁₃

^ H ■■; : ■; ■ ".:

^ CHOH (CH ₂ ) _e -CH ₃ XXi

009827/2053

in which m is a number from 1 to 6, ρ is a number from 0 to 7, η is a number from 1 to 8, a is a number from 0 to 4, b is a number from 5 to 7 and e is the number 6 or 7 is; is hydrogen iodide, an alkyl group having 1 to 4 carbon atoms inclusive, or a pharmacologically acceptable cation; Z is an ethylene group substituted by one or two fluorine atoms, methyl or ethyl groups, or by an alkyl group having 3 to 4 carbon atoms; Y is an isobutyl, tert-butyl, 3,3-difluorobutyl, 4,4-difluorobutyl or 4,4,4-trifluorobutyl group, and in which - the bond of the hydroxy, - (CH ₂ ) -COOR .., - or - (CH ₉ ) _m -Z-C00R ₁₅ group on the ring in ^ - or ß-position 'indicates. Each formula includes compounds in which the hydroxy side chain is in the R or S configuration.

Examples of alkyl groups with 1 to 4 carbon atoms '' including, are methyl, ethyl, propyl, butyl groups and their isomeric forms.

■ Pharmacologically acceptable cations of the formulas X to XXII, which fall within the scope of R. ^ , are quaternary ammonium ions or the cationic forms of a metal, ammonia or an amine. .

Particularly preferred are the cations of alkali metals, e.g. lithium, sodium and potassium, as well as of alkaline earth metals, e.g. magnesium and calcium, although cationic forms of other metals, e.g. aluminum, zinc and iron, too are within the scope of the present invention.

Pharmacologically acceptable amine cations in the context of R., in combinations of the! »Ormulas X to XXII derive from primary, secondary or tertiary amines. Examples of suitable amines are methylamine, dimethylamine, tri-diethylamine, Ethylamine, dibutylamine, triisopropylaminej N-methylhexylamine, Decylamine, dodecylamine, allylamine crotyl

009827/20 53

amine, cyclopentylamine, dicyclohexylamine, benzylamine, dibenzylamine, η -phenylethylarain, ß-phenylethylamine, ethylenediainine, diethylenetriamine and similar aliphatic, cycloaliphatic and araliphatic amines, which contain up to and including 18 carbon atoms, for example pyranolines, and also, for example, pyrolinoic amines, pyrrolic amines, and also morphocyclic amines, morphociperidines, β-phenylethylarain, ß-phenylethylarain, and furthermore, pyrrolic amines, for example, pyrolinoic amines , Piperazine, lower alkyl derivatives thereof, for example 1 - methylpiperidine, 4-ethylmorpholine, 1-isopropylpyrrolidine, 2-methylpyrrolidine, 1,4-dimethylpiperazine, 2-methylpiperidine and the like, as well as amines that are soluble in water or contain hydrophilic groups, for example mono-, di- and triethanolamine, ethyl diethanolamine, iT-butylethanolamine, 2-amino-1-butanol, 2-amino-2-ethyl-1,3-propanediol, 2-amino-2-methyl -1-propanol, tris- (hydroxymethyl) -aminomethane, H-phenylethanolamine, IT- (p-1ert-Amy! Phenyl) -diethanol amine, galactamine, IJ-ethylglucamine, H-diethy! Glucosamine, pheriylephrine, adrenaline, procaine and like that.

Examples of suitable pharmacologically permissible quaternary ammonium ions within the framework of R ,, in the formulas X to XII are eetramethylaniiaonium, tetraethylammonium, benzyltrimethylamracniuffijr phenyltriethylamraonium and the like.

The divalent · ethylene group Z is atoms on one of the carbon in the ^'y - or substituted ß abutment condition to the CarboxyIfunktion. For example, 2 is a -CHp-CIfF-, -CHP-. CH ₂ -, -CH ₂ -CP ₂ -, -CP ₂ -CH ₂ -, -CHP-CHE ¹ -, -5H ₂ -CH (CH ₃ ) -, -CH (CH ₃ ) ~ CH ₂ -, - CH ₂ -C (CH ₃ ) ₂ -, -C (CH ₃ J ₂ -CH ₂ -, or -CH (CH ^) CH (CH ₃ ) group, and the same applies to an ethyl group, ] to a fluorine and a methyl group "a J ¹ ItIOr and a ^ ethyl group and a methyl and an ethyl group. Z may also be on a carbon atom by a propyl, isopropyl, butyl, isobutyl, sec-butyl or tert-butyl-group substituted soin.

BADORIGINAU

9827/2053

Although all compounds of the formulas X to XXII die have particular advantages of a long-lasting effect and are administered orally, sublingually, intravaginally and reactally who can exist within this group of Compounds an even more limited group that share these properties to a particularly high degree. It is those links that form a chain of seven Have carboxyl terminated carbon atoms i.e. m = 4 and n = 6, especially those with total 20 carbon atoms excluding the branch, i.e.

. ρ = 4 and a = 1 when Y is a difluorobutyl or trifluorobutyl group a = 2 when Y is isobutyl and a = 3 when Y is a tert-butyl group. Regarding of the possible variations of Z are those most advantageous which have a fluorine or a methyl group, 2 fluorine atoms or 2 methyl groups on the same "carbon" atom or a butyl, isobutyl, scc-butyl or tert-butyl

. group on the carbon atom in c '- (adjacent) position to have the carboxyl function.

Of the compounds represented by Formula VIII the compounds of the closest formula are special Meaning:

¹¹⁰ ^ (CE ₂ ) g-CCOE ^

HC &quot; J ^ -

H CKCH- (CH ₂ ) ^ - CH ₃

in the R ₁ - and --- have the ver.stohnnä ar-e ^ level meaning. These connections are for those-of-pay for 5GF. _o the purpose of "explained .vert. but are surprising-.and \ unexpectedly enough, for this ZWC-ckc- useful than PGF., _y

It is known that PGF ^ _n . has one essential blood pressure ■ increasing V.'irkunc. That represented by Formula XXIII

009827/2053

--20 - ■■ '■■■■:. ■ .; ■ '.' ■■ -

.- ^ .- Hydroxylsomer, 8-IsO-PGF ₁ ^, has only a small fraction of the hypertensive effects of PGF. ^. The ß-Hydroxyisomere represented by the formula XXIII 'is, in contrast to PGF _1p, which is an antihypertensive agent ■ also οin mild Qb pressor agents. At the same time, these new compounds, namely 8-iso-PGF.,. and 8-isO-PGF. ο .sov / ie their salts and esters are essentially effective as nasal blood congestion-reducing agents, as agents for inhibiting platelet aggregation and as labor-stimulating agents for inducing childbirth. The new compounds of formula XXIII are therefore used instead of PGF ^ ₇ with obtaining unexpected benefits for these "purposes," since their cardiovascular effects are similar to, but significantly lower than, that of

, PGP ₁ /, -? & 3Ρή- _β and PG ^ ₁ as well as the other compounds of the formulas VII, VIII and IX, including the special compounds of the formulas X to XXIII, are used for the above corner in the form of the fröien acid, ie with R ₁ or; R ₁ ^ - = hydrogen, used in ester form or in the form of pharmacologically acceptable salts. Becomes an EE'ccr. is used, it can be any with R ₁ within the meaning given above. Preference is given to -je- -doclr'Esters with alkyl residues with 1:. "To _4 carbon atoms-: including. The methyl and ethyl esters are particularly preferred because of the optimal absorption of the compounds by the body or the test animal.

The pharmacologically acceptable salts of the compounds of the above formulas VII to XXIII, which are of value for the purposes described, include those which have cations within the meaning _{given above for - R 1} - "-";"."" ■. '■.. ^:

As already mentioned · /, the connections of the formulas

BADORlQiNAL

VII "to XXIII administered in different ways depending on the different uses, so ■ for example intravenous, intramuscular, subcutaneous, oral, intravaginal, rectal, sublingual, topical, as well as in the form of sterile ones Implants for extended effectiveness.

For intravenous injection or infusion, sterile aqueous, isotonic solutions are preferred. For this purpose, it is preferred because of increased water solubility that IL in the compounds of the formulas in the compounds of the formulas VII, VIII or IX, and R ₁ ~ ¹ X to XXIII is hydrogen or a pharmacologically acceptable cation. For subcutaneous or intramuscular injection, sterile solutions or suspensions of the acid, salt or ester in an aqueous or non-aqueous medium are used. For oral or sublingual administration, tablets, capsules and liquid preparations such as syrups, elixirs and simple solutions are used in conjunction with the usual pharmaceutical carrier materials. For rectal or vaginal administration, suppositories made according to ancestors known in the art are used. For tissue implants, a sterile tablet or silicone rubber capsule or some other material that contains the substance or is impregnated with it is used.

The compounds of the formula VII, including PGE ₁ , and the new compounds of the formulas X, XIII, XVI, XIX and XXII, the compounds of the formula IX, including PGA. and the new compounds of the formulas XII, XV, XVIII and XXI are prepared by new processes which are described below.

The compounds of the formula VIII, including PGF _1n , and the new compounds of the formulas XI, XVII, XX and XXIII are obtained by reducing the carbonyl group of the corresponding hydroxy compounds of the formula

009 82 7/2053

. '■■ ■''■"■■■ - ^{22 -:;":} ■ ■''■' ■: ■■ ^'■■■; - ■■■■ '"■ ■' :. '

VII, including PGE. and the new compounds of the formulas X, XIII, XVI, XIX and XXII. In the case of the conversion of PGE .. to a mixture of PGF .. and PGF _{1 ß} , this reduction process is known; see, for example, Acta Chem. Scand., Vol. 16, p. 969 (1962) and J. Biol. Chem., Vol. 239, p. 4101 (1964). The other compounds of the formula VII, including the new compounds of the formulas X, XIII, XVI and XXII, are reduced by the same process with sodium borohydride to give the corresponding t -'-. and β-compounds of formula VIII, including the new compounds of formulas XI, XIV, XVII, XX and XXIII.

The compounds of the formula IX, including PGA ₁ , and the new compounds of the formulas XII, XV, XVIII and XXI are prepared by dehydrating the corresponding hydroxy compounds of the formula VII, including PGS ₁ , and the new compounds of the formulas X, XIII, XVI, XIX and XXII, ■. made. This process is known for the case of converting PGE ₁ to PGA _1; see, for example, Biochem. Biophys. Res, Commun., Vol. 21 _p . 413 (1965) and Pike et al., 'Proc. Hobel Symposium II Stockholm (1966) Interscience Publishers New York, pages 162-163 (1967). The other compounds of the formula VII, including the new compounds of the formulas X, XIII, XVI, XIX and ZXII, are dehydrated, for example with aqueous acetic acid, to give the corresponding compounds of the formula IX, including the new compounds of the formulas XTI, XV, XVIII and XXI.

According to a further subject of the present invention, compounds of the E ₁ series, ie compounds of the formula VII in which R _{1 is} not hydrogen (hereinafter referred to as R ₇ instead of R ^), and compounds of the A ₁ series, ie compounds of the formula VII, in which R _{1 is} not hydrogen (hereinafter R ~ instead of L) after a new reaction sequence according to

^ BADORIQINÄL

-23 -
Scheme Ä PC _n H _2n -COOR ₇ ^ C _n H _2n -COOR ₇ H CR ₄ -CR ₂ R ₃ !
ι
CR ₄ ⁼ CR ₂ R ₃

XXIV

XXV

"C (OH) R ₄ -C (OH) R ₂ R ₃

XXVI

C _n H _2n -COOR ₇ H

C (OSO ₂ Re) R ₄ -C (OSO ₂ R ₆ ) R ₂ R _:

XXVI I

HO '

Η '

Vl I

- ^ C _n H _2n -COOR ₇

H'

IX

RS R ₂

009827/2053

- -24 -: ■■ - ■,

In scheme A, R ₂ , R ₃ ^ R ₄ , C _n H _2n and ^ have the meaning given above. R ₇ has the same meaning as R ^, but exclusively hydrogen. Rg is an alkyl radical having 1 to 5 carbon atoms inclusive. The reactants XXIV, XXV, XXVI and XXVII are all in the exo configuration with regard to the -CR _A = CR _? R ^ -,

-CR ₄ -CR ₂ R ₃ -, -C (OH) R ₄ -C (OH) R ₂ R ₃ - and -C (OSO ₂ R ₆ ) R ₄ -C- (OSO ₂ Rg) R ₂ R ₃ -Groups. .

Scheme A also shows the transformation / end products of the formula VII to end products of the formula IX. As above he * - this transformation is in the case of PGE., (VII) and PC-A. (IX) known and does not form part of the invention.

The starting materials, ie the Olefin XXIV and Epoxy XXV, are known: see Belgian Patent No. * 702.477, as a special print in Farmdoc Complete Specifications _s Vol. 714, Mr. 30.905, p. 313, March 12, 1968, ■ .. '*'.

According to this Belgian patent, the reaction sequence is after which the olefin XXIV is obtained as follows: The Hydroxy group of 3-cyclopentenol is, for example protected by a tetrahydropyranyi group, content filament : v / ird a diazoacetic acid ester to the Böppilbindting

being an exo-endo mixture of a bicyclo- "3 * 1'-ÖJ ^ -hexane arises, in the 3-position by the protected hydroxyl group and in the 6-position by an esterified Qarbpxylgi'uppe is substituted. Las exo-endo mixture is made with a Base treated to further add the endo-isomer in the mixture isomerize exo isomer. Now the ester group in the 6-position i, n an aldehyde or keto group, namely O

-CHO- or -OiO ',' converted. Subsequently, this aldehyde or keto group is ucieetzt by a Wittig reaction to a group of the formula -CR ₄ = CR ₂ R ₃ , which is in the exo configuration to the bicyclic ring structure and is the same,

009827/2053 BADO _RJGJNAL

- 25 -

as indicated above in Formula XXIV. Then, the protective group is removed to re- around the 3-hydroxy ^group: generate, which is then oxidized for example, by Jones reagent, .to give a compound of the following formula: ■

XXVIII

in the r _? , R ₇ and R, which have the above meaning, with an exo configuration in view of the -CR. = CRpR ^ group .. Finally, the compound XXVIII with a * - * / - halogen ester of the * formula Br-C _n H _2n -COOR ₇ or 1-C _n H _2n -COOR ₇ alkylated to give an olefin XXIV, where CH has the meaning given above and the -CH ₂ -COOR ₇ -GrUpPe in: \ .- or ß-configuration on the Cyclopentane ring is bound.

There are four isomers of Olefin XXIV, excluding the enantiomeric forms, which double that number. There are cis and trans forms with respect to the -CR ^ = CRpR, group, and each of them can have i »^ or β-positions with respect to the -C _n H ₂ -COOR ₇ groups . In the aforementioned Belgian patent no. ~ 7-2477 the preparation of each isomer is described. The cis and trans isomers of the unalkylated ketone XXVIII are separated in this step, and the separated cis and trans isomers are each alkylated to give a ""., And ß-mixture of the olefin XXIV, of which the ',> - and ß-isomers are separated. On the other hand, the 'cis-trens mixture of the formula XXVIII can also be alkylated to a mixture of four isomers of the olefin XXIV, namely c \ -eis-, - ¹ X-trans, β-cis and β-trans isomers, and the isomeric components of this mixture are separated from one another, or the mixture is used as such.

If the olefin XXIV according to scheme A is to be converted into PGE ₁ -EStGr or PGA ₁ -Ester by the new process according to the invention

009827/2053

are changed, so are in the olefin XXI? R-, and R, hydrogen, R ₂ pentyl, C _n H _2n hexamethylene and the -C _n H _2n -COOR ₇ group is bonded in the C 'configuration. The 8-iso-PGE ..- esters or 8-iso ^ PGA ..- esters are made from the same olefins, but in which the -CIL, -COOR ₇ -GrUpPe is bound in the ß-configuration. In order to produce a dious group of olefin esters, Br- (CHp) ₆ -COOR ₇ or I- (CHg) ₆ -COOR ₇ for the alkylation of XZVI to XXIV _{7 is} required, and hexyl bromide to produce the necessary Wittig reagent, for example hexyltriphenylphosphonium bromide. These intermediates are known or are prepared by known processes. The other Wittig reagents which are required to prepare the radicals ' ■■ -CR. = CRpR are prepared from known compounds or by known processes. The other various '-' halogen esters which are required to produce the ReStG-C _n H _2n -COOR ₇ are also known or can be produced by known processes.

In order to explain these intermediates, the specific compounds of Ponaeln X to XII are used. The olefins of the formula XXlV, which are necessary as reactants for the preparation of compounds of these formulas, require the following halides as reagents, with the aid of which the necessary Wlttig reagents are prepared, namely CH, - (OE ₀ ) ~ CH ₀ -] L and T- (CH ₀ ) -CH ₀ -X., Where X, Y, a and ρ have the meanings given above. The CH, - (CHp) -CHp-X-halides are produced by reacting the corresponding primary alcohols, all of which are known, with PBR or PCl ₇ . The Y- (CH ₂ ) _ä -GH ₂ -X compounds,

where Y is a (CK ^ IpCrI-CH ₂ - or (CH,) ^ CH group are prepared in the same way from the corresponding alcohols. The low molecular weight alcohols, for example (CH ₅ J ₂ CHCH ₂ CHpOH and (CH ₃ ), CCH ₂ CH ₂ OH, are known. The remaining alcohols are converted by reaction

009827 / 20S3 BADORIQiNAL

of these prior art alcohols corresponding bromides with sodium cyanide, hydrolysing the nitriles produced obtained to the corresponding carboxylic acids _v and reducing these acids to the corresponding primary alcohols by lithium aluminum hydride, whereby each time .the (CH ₀₎ is extended around a chain .Kohlenstoffatom until all Bromides are produced.

The Y- (CH ₂ ) -CH ₀ -X compounds in which Y is 3,3-difluorobutyl are _{prepared from ketocarboxylic acids CH 7} -CO- (Cm), - COOH, where d 2, 5, 4, 5 or 6 is. All of these keto acids are known. The methyl esters are prepared and reacted with sulfur tetrafluoride to. to cool the corresponding CH ₅ -CF ₂ - (CH ₂ ) _d -COOCII - compounds, which are then reduced with lithium aluminum hydride su CiU and then ni "t?! DBr, odor PCI, to the compounds CH ₅ -CP ₂ - ( CK ₂ ) ^ - CiL-X are unrcetct. Lic compounds Y- (ClU) _n -CH ₂ -X, b-ö-i where Y is -in 4,4-difluorobutyl, are from the known carboxylic acids HOOC- (CH ^) ^ "COOH, where f is 3 >> 4, 5, 6 or 7. These dicarboxylic acids are _{esterified to CE 2} OGC- (CH ₀ ), -COOCH, and then _{semi-saponified, for example with ßarinialiydro: cyd t} , to give lic.lb * 'ester 1100, C- (CH ₀ ) ^ - COOCH ₅ . The free carboxyl group is first converted into the acid chloride with thionyl chloride and then converted into an aldehyde group by Rosennund heduction. By reaction of the alcohol with Srhwpfplt fluoride one obtains CHF ₀ - (CH ^) "- COOCH ^, which after successive treatment with lithium aluminum hydride and PBr, or PCI, which results in _{the required CHF 2} - (CH ₂ ) _{f -X.} The Y- (CH ₂ ) _a -CH ₂ -X compounds in which Y is 4,4,4-trifluorobutyl are _{prepared from aldehydes CH 5} OOC- (CH ₂ ) ^ - CHO, which are as given above would have been received. Reduction of the aldehyde with sodium _{hydride gives the alcohol CE 5} OOC- (CH ₂ ) ^ - CH ₂ CH. The reaction with PBr or PCl ₅ gives CH ₅ OOC- (CH ₀ ) _f -CH ₂ -X. The saponification of this ester gives the carboxylic acid, which is

009827/2053

- 28 - - - ■ - ■. ■:.

Settlement with sulfur tetrafluoride _{gives the required CF 7} - ■ (CHp) 0-CH ₂ -X. The * reactions with Si \ are described in U.S. Patent 3,211,723 and Dem-J.-Org.Chem., Vol. 27, p. 3164 (1962).

To prepare the olefins of the formula XXIV ₈ from which the special compounds of the formulas X to XXII are prepared, the '' - "'- bromides and iodides of the formulas Q- (CHp) Z-COOR ₁ .and / Q- ( CH ₂ J _{n -COOR1.} necessary where Q is Br or I, R ₁ is. an alkyl radical having 1 to 4 carbon atoms, inclusive, and Z, m and η which are as defined above. the compounds of the formula Q- ■ (CH ₂₎ -COOR. ₁ with Q = bromine are known or are finally reaction with PBr prepared from known dicarboxylic acids half esters by reacting the carboxyl group with thionyl chloride to the acid chloride, reduction with sodium borohydride to an alcohol and *. the iodide is achieved by treatment of the bromide with sodium iodide in acetone. The compounds Q- (CHg) _1n -Z-COOR ₁₄ are prepared using the corresponding succinic acid, HOOC-Z-COOH> in which Z has the meaning given above, and are all known. to the anhydrides converted and reacted with an Alkanol.-JL. ^ OH, whereby one .sowohl the HOOC-Z-COOB _{1 4} -alsauch the R ₁ ,, OOC-Z-COOH isomer obtained. The "free carboxyl group is then converted into the acid chloride with thionyl chloride, this is converted into an aldehyde by Rosenmund reduction, the latter into an aldehyde with sodium corhydride and this _{into the eromide with PBr 7} , whereby Br-CHp-Z-COOR ₁₇ , or R ₁ -OOC-Z-CKp-Br. This brings the necessary substituents of Z into the correct ratio to the -COOR _{1 group} . Then the -CHp- group is added as often as necessary by adding -Br is replaced by -CN (sodium cyanide), -CN is hydrolyzed to -COQH and -COOH is converted, as described above, Iu-CH ₂ Er .'with sodium iodide in acetone. '-_'-

, ■■ '■ ... ": - -'. _ - ■■ .- '■''; .. ■■ ..'^■:; : badorksimäl ,. '. ^: '., .' ■■■■ '■ *.

009827/2053;.-.-..- ^: -> ^: ■■ * - ■ - ^ J.

By equivalent processes, which are all known, the person skilled in the art can use all halogen esters and Wittig reagents which for the preparation of all olefins within the scope of the formula XXIV are necessary.

Scheme A also shows the conversion of the olefin XXIV to the epoxide XXV. This reaction is in the Belgian mentioned U.S. Patent No. 702,477; it is made by reacting the olefin XXIV with hydrogen peroxide or a percarboxylic acid, for example m-chloroperbenzoic acid or perlauric acid, carried out. This stage does not form part of the invention.

The conversion of the olefin XXIV to the Grycol XXVI is carried out by reacting the olefin with a hydroxylating reagent. Hydroxylating Reagents and Methods for this purpose is known; see, for example, Gunstone, Advances in Organic Chemistry, Volume 1, Pages 103-147 (1960), Interscience Publishers, New York. From the C ^ -cis form of the Olefins XXIV are obtained with the help of a cis-hydroxylating By means of, for example, osmium tropoxide, two isomeric c <_- ery ~ thro-glycols of the formula XXVI, and from the cx-trans form des Olefins XXIV, on the other hand, uses the same cis-hydroxylating agent to produce two isomeric threo-glycols of the formula XXVI. Similarly, the ß-form of olefin XÄIV gives two isomeric ßerythro-glycols of the formula XXVI, using the same cis-hydroxylation agent, and the ß-trans form des Olefin XXIV two isomeric ß-threo-glycols of the formula XXVI. These-c-erythro, ol-threo, ß-erytrho and ß-threo isomer pairs of glycolon are determined by silica gel chromatography separated into individual isomers, namely a more polar and a weaker polar isomer.

The conversion of the epoxide of the formula XXV to a glycol XXVI (see scheme A) is carried out by reacting the epoxide with an acid, the pK value of which is less than 4. Examples of such acids are formic acid, chloroacetic!

009827/2053 '

- 30 -.,: ■ "■ :, ■:, '■; V

acid, dichloroacetic acid, fluoroacetic acid, trifluoroacetic acid, Oxalic acid, maleic acid and the like. Formic acid is particularly preferred .. Usually it is sufficient that Keeping the epoxy reaction mixture at about 25 ° C only for 10 to 100 minutes. The glycol ester that received is then, advantageously with a weak base, e.g., ilodium bicarbonate which Glycol XXVI hydrolyzes to.

According to scheme A, the glycol XXVI by reaction of XXVI with an alkyl sulfonyl chloride or bromide or with an alkanesulfonic acid anhydride into the corresponding bis-alkanesulfonic acid ester of the formula XXVII, where the. Alkyl radical; respectively. 1 to 5 carbon atoms inclusive e-holds. Alkyl sulfonyl chlorides are preferred for this reaction. The reaction -will "in the presence of a base for neutralization the acid formed as a by-product. Particularly suitable bases are .die.-tertiary amines, e.g. dimethylaniline or pyridine. It is enough in general off to mix the two reactants and the base and the mixture for several hours in a temperature range from 0 to 25 C. The bis-sulfonic acid ester of the formula XXVII is then isolated by known methods.

As shown in Scheme A, the conversion of the bis-, sulfonic acid ester XXVII is carried out to the final product VII by reacting with water. · This reaction is carried out by mixing the compound XXVII, with water at about O to about 60 ⁰ G. For the production of PGE ₁ or 8-iso-PGS. a temperature of 25 ° C is usually sufficient and conversion is complete in about 5 to 10 hours. A homogeneous reaction mixture is advantageous. It is obtained by adding a sufficient amount of a water-soluble organic diluent which does not take part in the reaction. Acetone is a suitable diluent. The desired product is obtained by evaporating the excess water and adding

009827 / 2O53

if diluent is isolated. The residue contains a Mixture of isomers of the formula VII, which differ in the configuration of the hydroxy rare chain (R or / S). They are by the by-products - and by one another Silica gel chromatography separated. "A common by-product is a monosulfonic acid ester, corresponding to the bisulfene acid ester of the formula XXVII with the difference that the -OSOpRg group to the. the carbon atom adjacent to the cyclopropane ring of this formula is replaced by -OH. This monosulfonic acid ester is made in the same manner as above • Described for the conversion of the glycol XXVI to the bis-Estßr XXVII, esterified to the bis-sulfenic acid ester of the formula XXVII and in this way additionally supplied to the end product of the formula VII. :

During the conversion of the Bi3 ester XXVII to the end product VII is preferably the bis-mesyl ester, i.e. a compound XXVII, in which Rg is methyl, is used.

The configuration of the -C _n IL ^ _n -COOR - group ir. Α em bis-3ster XXVII does not change during the conversion of XXVlI to VII. If, therefore, in formula X] -VII R "pentyl, R ^ and R, - ^ vas ^ erstoff and CK ₉ hexanemethylv.li rind, 1-GE esters are obtained if the - (CH ₉ ) ₆ C00R ~. -Grupp <2 is initially bound in the i -configuration, and 8-1so-PGE.-ester when the - (CH _? ) ^ C00R, ~ group was initially in the "configuration. Both erythro isomers and both thrc-o-Tporaeren of the bis-2nd of the CX compound of the formula aXVII, however, give the same Pro-. duct of the I ^s compound of the Foririe.l VIl In essentially the same yield; the same applies to the B connections. 'Dalier has the AusgangsmafcorJal <fex · üOrmel are not separated in cis and trans isomers XXIV, and it is not necessary that d \ irch the hydroxylation of XXIV to the glycolic col XXVI received vei'schiedenon erythrp- and; to separate threo isomers. In other words, all suffices of erythrothreo isomers of the formula XXVII for the production of the "" Etict-

009827/2053

- 52 - 7 ■. "■■■;: ■■ -.. ■;

Product YII the same, valuable and just as useful as any single isomers.

As shown in Scheme A, the conversion of the bis-sulfenic acid ester XXYII to end product IX is carried out by heating XXVII within a temperature range of 40 ° to 100 ° C. with a mixture of water, a base, which is adjusted by a pH in aqueous solution is characterized from 8 to 12, and a sufficient amount of an inert water-soluble organic diluent to obtain a basic and substantially homogeneous reaction mixture. A reaction time of one to 10 hours is usually used. Preferred bases are the seriously borrowed salts of carboxylic acid, especially alkali metal bicarbonates, for example sodium bicarbonate. A suitable “diluent is acetone. The products are isolated and separated in the manner described above for the conversion of the bis-ester XXYII to the end product VII. The same monosulfenic acid ester obtained as the Hebetiprodukt Final product IX '. As / in the production of YII, the ice mesyl ester of XkVII is preferred also in the production of IX. Just like making: YII precursors making IX ^ X-XXVII makes / X-IX ^ MXVII makes ß-IX, all erythro and threo isomers of .IC ¹ ITII are equally useful in making IX, and in In each case, with - ^ - IX like .ß-IX-, a mixture of R and S is oraer s is obtained. These R 1 and S isomers are separated by silica gel chromatography.

It should be noted that all compounds XXIV, XXV, XXVI and XXYII are in the exo configuration. Unerv / art et evidence has found that significantly higher yields of the end product YII are obtained when the bis-sulfonic acid esters are in the endo configuration instead of in the exo configuration with regard to the -C (OSO ₂ R ₆ ) R _/ - C (OSO ₂ Rg) R ₂ R_ group present. "These endo-compounds are after the same

009827/2053 BADORlQiNAL.

Process prepared as described above and in the aforementioned Belgian Patent No. 702 477 for the corresponding exo compounds / has been described, apart from the Use of the mixture of exo- and endo-bicyclo / 3,1,0'-hexane, that in 3-position by the protected hydroxy group, e.g. tetrahydropyranyloxy group, and in the 6-position by an esterified carboxyl group mentioned above as an early intermediate and prior to further use essentially completely to the exo-Porm was isomerized. Instead of this exo-endo mixture becomes the corresponding -pure endo-isomer as an intermediate used. This endo configuration is then used during the subsequent transformations described in the Belgian patent are described, retained, and one obtains the olefin XXIV and epoxide XXV (Scheme A) and the glycol XXVI and the Bis-sulfenic acid ester XXVII.

The required pure endo intermediate from lOrinel

XXIX COOCH,

is prepared by endo-bicyclo-J5,1, Öj-hex-2-en-6-carborifiä-iii-omfthy1 udder with diborane in a mixture from tetrahydrofuran and diethyl ether. This reaction is known and gives the enrio-Moyol.n - / 3,1, υ / - li <=> van-3-01-6-carboxylic acid methyl ester, which is then reacted with dihydropyran in the presence of a catalytic amount of POCl to give the desired prebinding of the formula XXIX. This connection is then used, as described above, to finally to produce the endo-isomer of all compounds and isomers of the bis-esters of lOrmel XXVII.

'.00982 7/20 5 3

The process for converting the ehdo isomers of the bis-sulfonic acid ester XXVII in, the end product VII and the results of this conversion, i.e. the isomerism of the compound of Formula XXVI and the product of Formula VII are the ... - ■ same as above for the conversion of exo-XXVI to VII were described, apart from the fact that the yield of product VII from endo-XXVII was quite unexpected is much larger than from exo-XXVII.

These final products of the formulas VII and IX which have been prepared as described above, all R = ₇ ester, w in which R ₇ has the meaning described above. For some of the purposes explained above, it is preferred that the compounds of the formulas VII and IX are present in the form of the free acid or as salts, for which the free acids are required as starting materials. the esters of formulas VII and IX are difficult to hydrolyze or saponify occur without undesirable structural changes in the desired acids. Three other methods are of -Viert to prepare the products of formulas VII and IX in the form of the free acid.

One of these processes can-predominantly- be used for the preparation of the free acids from the corresponding alkyl esters in which the alkyl group contains 1 to 8 carbon atoms inclusive. This process consists in subjecting the allyl ester of the formula VlI or IX to the acylase -. " Enzyme system exrieis microorganism of the species Subphylum 2 from Phylum III and then isolating the acid. The species of the genera Mucorales, Hypoereales, Moniliales and Actinomycetales. Furthermore, species of the families MuGoraceae, Cunninghamdellaceae, Nectreaceae, Miniliaceae, Dematiaceae, Tuberculariaceae, Actinomyeetaceae and Strepto- ■ \ mycetaceae are particularly preferred for these purposes

009827/2053 BADORlQiNAU

- 35 - ■. - -. ■■ "■ ':

Purpose species of the genera Absidia, Circiriella, Gongronella, Rhizopus, Cunninghamella, Calonectriä, Asperglllus, Penicillium, Sporotrichum, Cladosporiuin, Pusarium, Noeardia and Streptomyces particularly preferred.

Examples of microorganisms that are within the scope of the preferred Classes, tamils, and genera are covered in US patent No. 3 290 226 listed.

The enzymatic ester hydrolysis is effected by shaking the allyl ester, of the formula VIT or IX in aqueous suspension "with the enzyme contained in a culture of one of the microorganisms mentioned above, until the ester is hydrolyzed. A" reaction ion temperature ranging from 20 ° to 3O ⁰ C is generally satisfactory. A reaction time of from one hour to 20 hours is normally sufficient to effect the desired hydrolysis. Exclusion of air from the reaction mixture, for example by means of argon or nitrogen, is usually desirable.

The enzyme is obtained by separating the ropes from the culture, the washed and resuspended in water and then, for example by stirring with glass balls or by Sonic or ultrasonic vibrations ^ are crushed. That The entire aqueous genic is then used as a source for the Ensym used. Preferably one can also remove the cell residues Centrifuge or filter and remove the supernatant solution

or use the Piltrat *

In some groups it is advantageous to protect the microorganism against an alkyl ester of an aliphatic acid the acid including 10 to 20 carbon atoms and the alkyl group has 1 to 8 carbon atoms inclusive contains, or such an ester to the KuI-. door and the culture 1 to 24 hours before the line, "separation without: holding further growth. Hieidttreh yrird ^

aG9827 / 2053

sometimes the enzyme is more effective in converting the ester VII or IX to the free acid. An example of an alkyl ester for this purpose · is methyl oleate.

Enzymatic hydrolysis is general to conversion from prostaglandin alkyl esters to the free acids and therefore not only used for the production of the free acids Alkyl esters of formulas VII and IX useful, but also for Conversion of other known prostaglandin alkyl esters and their analogues, for example the alkyl esters of the formula VIII as well as the esters of prostaglandins, such as PGEp,

PGAg, PGA ₇ and the like; See Bergstrom, among others (see above), 'as well as reference points, the v noted above for other known Prostaglandinalkylester / ground, which can be hydrolyzed by this enzymatic method.

Although, as mentioned above, the esters of Formulas VII and IX are not easily hydrolyzed and saponified to the corresponding free acids of Formulas VII and IX, some of these esters can be converted to the free acids by a different method. These include the halogen ethyl esters, in which IL is in the ß-position by 7>. Fihlöratome, 2 or 3 bromine atoms or 1,2 or 5 iodine atoms is substituted. Such esters in which, for example, R ..-. CIUCCl- is converted into the free acids by treatment with zinc metal and an alkanoic acid with 2 to 6 carbon atoms, preferably acetic acid. Zinc dust is preferred as the physical form of zinc. Mixing the halogenoethyl ester with the zinc dust at about 25 ° C. for several hours usually results in an essentially complete replacement of the halogenoethyl group of the ester. Formula VII or IX caused by hydrogen. The free acid is then isolated from the reaction mixture by known methods. ·

009827/2053 "

ORiQl INSPECTED

As shown in Scheme A, the haloethyl esters of the formula VII and IX are obtained from the corresponding bis-sulfonic acid esters of the formula XXVII, where R _{7 is} in the β-position by 3 chlorine,

2 or 3 bromine or 1, 2 or 3 iodine atoms, preferably

3 chlorine atoms, is substituted. These conversions are carried out as described above for the other reactions from XXVII to VII and XXVII to IX.

The bis-sulfonic acid esters of the formula XXVII, in which R ₇ is substituted in the β-position by 3 chlorine, 2 or 3 bromine or 1, 2 or 3 iodine atoms, are obtained from the corresponding glycols of the formula XXVI according to the above for the others Conversions of Formula XXVI to Formula XXVII prepared procedures described. -

The glycols of the formula XXVI, in which R is substituted in the β-position by 3 chlorine, 2 or 3 bromine or 1, 2 or 3 iodine atoms, are obtained by hydroxylation of the corresponding olefin ^{β of} the formula XXIV or Epoxydg of the formula XXV according to the above for the other conversions from XXIV to XXVI and

XXV obtained according to the method described in XXVI. These haloethyl esters can also be prepared by esterifying the free acids of the glycol of the formula XXVI (where R _{7 is} hydrogen) with the corresponding haloethanol, for example β, β, β-irichloroethanol, if the haloethyl group is to be -CHpCCl-. This esterification is carried out by reacting the free acid of the formula XXVI with the haloethanol in the presence of a carbodiimide, for example dicyclohexylcarbodiimide, and a base, for example pyridine. From this mixture is obtained, namely, advantageously with an inert diluent, for example dichloromethane, usually the desired Halogenäthylester within several hours at about 25 ⁰ C. The required for this esterification Glyeolsäuren of formula

XXVI are obtained by hydroxylation of the free acids of the olefin of Formula XXIV by the procedure described above for the other transformations from XXIV to XXVI. "

^ 009827/20 53

- 38 - : ■■: ■■ ν .. ■■;

The olefins of the formula XXIV, in which R _{7 is} substituted in the J3 position by 3 chlorine, 2 or 3 bromine or 1, 2 or 3 iodine atoms, are esterified with the corresponding halogenoethanol, for example CCl ^ CHpOH, according to the above for the esterification of the glycolic acid of the formula XXVI (R ₇ = H) described method.

The required free acids of the olefin of the formula XXIV are produced by saponification or hydrolysis of the corresponding esters. However, this reaction is difficult without partial isomerization of the Cv. Isomer to the β-isomer or

w) - to carry out the B-isomer to demiX-isomer. It is therefore preferred to reduce the ring carbonyl group of an olefin ester of the formula XXIV to a hydroxyl group with sodium borohydride and then to saponify the ester. The latter to--. Settlement takes place easily and without isomerization. The hydroxyolefin obtained is then oxidized back with a free carboxyl group to give the ketoolefin of the formula XXIV (R ₇ is now hydrogen). For the latter oxidation it is necessary to use a reagent which does not change the -CR ^ = CRpR-ZG group of the compounds of the formula XXIV. The Jones reagent is suitable for this oxidation, see J. Chem. Soc, London, Volume 59, (1964) * All three conversions,

. namely the sodium borohydride reduction, saponification and oxy- ■ · dation, are carried out according to general methods known to the person skilled in the art. ;

During this second way to obtain the free acids of formula VII and IX based on the exo compounds shown in Scheme A. has been explained, the method can also be applied to 'the corresponding endo compounds.

In a third process for the preparation of the free acids • 'of the formula VII, a ketal of the following

f '

- 39 - ■ '

the following formula is used:

R ₁₂ O.

(J »an ^ _8..

- XXX

in which R ₂ !, ^R 3 »^ 4 ¹ ^ ^C n ^K 2n ^{aie have the meaning} given above, Rg hydrogen, an alkyl radical with 1 to 8 carbon atoms including," cycloalkyl "with 3 to 10 carbon atoms including, aralkyl radical with 7 up to 12 carbon atoms inclusive, phenyl or one with 1 to 3 chlorine atoms or with aikyl with 1 to 4 carbon atoms including substituted phenyl radical, R ₁₂ both an alkyl radical with 1 to 6 carbon atoms inclusive or, if they are bonded to one another, 1,2-alkylene- or 1, 3-alkylene groups with 2 to 6 carbon atoms inclusive, and-> a bond of the -C Hp -COORg- group to the ring in iX- or ß-configuration and exo- or endo-configuration with regard to the -CR. = CRpR, group. Such ketals, in which both Keste'R ^ denote an'Alkylrest, are prepared by a ketoolefin of the formula XXIV (R ₇ is Rp, as explained above) in the exo- or endo- Configuration in view k to the -CR _-1 =

CRpR-z group with an orthoformic acid ester of the formula HC (OR .. ο) · =; converts, where R ₁ ρ has the meaning given above. If the R ^ groups bonded to one another are 1,2-alkylene or 1,3-alkylene groups, the same ketoolefin of the formula XXIV is made with a 1,2-C-lycol or a 1,3-glycol having 2 to 6 carbon atoms inclusive in the presence of a strong acid, especially a sulfonic acid, e.g. p-! coluenesulfonic acid. Examples for,

en

■ (1,2-alkyl groups with 2 to 6 carbon atoms "are

- -CH ₂ CH ₂ -, -CH ₂ -CH (CH ₃ ) -, -CH (OH ₅ ) -CH (CH ₃ ) ^, -C (CH ₃ ) ₂ -CH ₂ -,

- -C (CH ₅ ) ₂ -C (CH ₃ ) p and -CH ₂ -CH (CHp-CH ₃ ) -. "Examples of '

009827/2053

1,3-Alkylene groups with 3 to 6 'carbon atoms inclusive are -CH ₂ CH ₂ CH ₂ -, CHp-CH (CH-J-CH ₂ -, -CH (CH ₃ ) -CH ₂ -CH ₂ -CH ₂ - , -CH (CH ₃ ) -CH (CH ₃ ) -CH ₂ -, 'and -CH ₂ -C (CH ₃ > _?. CH ₂ -. Examples of 1, 2-Glycols and 1, 3-Glycols correspond to the above examples fiir _r 1, 2-alkylene and 1,3-alkylene groups with an -OH group at any free valency. These two methods are generally known to the skilled person. '. ■

In scheme A, the ketal of the formula XXX becomes a ketal of the formula VII, ie ψ of the following formula, via the ketals which correspond to the epoxide XXV, de: a G-lycol XXVI, the bis-sulphoil acid ester XXYII converted: ■ - .. "■

. - ^ - ₀ OR ₁₂ -. -

. '■■■ Ho'^': ° "G ^ xxxi

in the. R ₂ , E ^ j R ^ ₇ ? I _? , R. ", C Ii ₂ and - ^ the. ■ stated above; Have meaning ». These instructions will be carried out. .- ■ leads, As described above for the conversions from XXIV to -. XXV, XXIV after IXVl, 'tXJ after XXVI ^ XXVI after X:; VII and for XXVlI to VII- besciiriebftn vmr.de-, with the difference that ketal glycol XXVI present as free acid is esterified before: - - · the conversion zum, lCetal-bis-S.ul.fon.säui-oc ;; Ucr XXVTT, xmä. except that several ketals of the formulas XXl '/, XXV, XXVI and XXVII are either in the exo or end ^ configuration rather than merely in the ezo configuration, as shown in Scheme A. . . ". '■■..'

The Zetal XXXI is now: according to known methods to a -. ■ "■"'. free acid fr. ^ = !!) saponified and. then with an acid, for example, ¹ ■ oxalic acid, hjrdrolysiert, give a final product VII 'is in cem R _7, V / asserstofi. '-

; ^: ■■ -. ■■ '■■■■: ^v . ■ _: /..-- '-Q ^ vS-B- 2 7/2053 ■ ■.''' / '' ■ ': BADORIQiNAU'.:;".

These ketal reactions are valuable for the preparation of compounds of the formula VII in which R _{1 is} hydrogen when the -C EL -COORo group is bonded in either the i ^ - or the B configuration. If R, and R, are hydrogen, CH is hexamethylene and - (CH ₂ ) ^ - COORo inc ^ -configuratioii, PGE ₁ (R and S) is obtained. If R and R are hydrogen, C _n II is _2n hexamethylene ,, and the - (QILjg-CQORg group in ß-configuration, one obtains 8-IsO-PGE ₁ (R and S).

In the in Belgian Patent 702,477 Nr- method "described for the preparation of olefin -XXIV is usually obtained a mixture of / X and .beta.-isomers with respect to the -C Vv ^r ie _n H _2n COOR ^ group. Above 'As described, these two isomers lead to the formation of compounds of the formula VII (X) and to compounds of 8-iSo- ₁ (β) .. If one of these types is preferred, there are two processes by which To favor the preparation of the preferred endisomers of the formula VII;

One of these methods is the isomerization of the final product d'er formula VII, in which R ₇ . Has the meaning given above or is hydrogen. VII of the formula either dasCTv isomers or. Beta isomers is held in an inert liquid diluent in a range of 0 ° to 80 ⁰ C and in the presence of a base, which is characterized by a pH value of their wässirlgen solution below about 10, until a substantial amount of one isomer has been isomerized into the other isomer, ie. Preferred bases for this purpose are the alkali metal salts of carboxylic acids, particularly alkanoic acids having 2 to 4 carbon atoms, for example sodium acetate, examples of suitable inert liquid diluents are alkanols having 1 to 4 carbon atoms, for example ethanol .. This reaction takes place at about 25 ⁰ C while about 1 to about 2.0 days. Apparently an equilibrium is being established. In the case of ron PGE ₁ and B-iso-KJE .. the equilibrium leads to 9 parts PGiL and

09827/2053

BATH

one in part 8-iso-PGEi- ". The mixtures of the two-γ- and" B-isomers are separated from the reaction mixture according to known methods, and then the two isomers are separated according to known methods Methods, for example by chromatography, recrystallization or a combination of these separately. The less preferred isomer now becomes the same Subjected to isomerization to the more preferred isomer to recover. In this way, essentially all of the amount of the less preferred isomer of the compound becomes of formula VII converted to the more preferred isomer by repeated isomerizations and separation processes.

The second method of promoting the manufacture of a beloved : ■ '■ preferred Endisoraeren of the formula VTI works.mit the olefin XXIV. Either the: / - or the ß-isomer of the olefin of. ■. "..- ■■ Formula XXIV is converted into a mixture of both isomers -. ; by keeping the ■ '■■ -> - or ß-isomer in an inert

liquid diluent at 0 to 10O ⁰ C in the presence. a base in the range until a substantial amount of the 'starting isomerone has been isomerized to the other isomer. Preferred bases for this isomerization are alkali metal arfiids, alkali metal alkoxides, alkali metal hydrides, and. '"' - ■■ TriarjT-imethylalkalim.etalle .. Al-. ■ Kalimetall-tert-alkOzide with 4 to 8 carbon atoms, for example potassium-tc-rt-butoxide. This reaction proceeds at about .25 ''C continued quickly ("1 minute to several hours). Obviously, starting from one or the other isomer, an equilibrium mixture of the two isomers is formed. . In the case of an olefin XXIV, in which Rp is pentyl,. -R ^ and .R, hydrogen, R _{7 is} methyl and C EU _Ί Ilexamethylene, the equal / weight mixture contains about ".1 / 3i>'- ^ - isomer and 2/3 ß-isomer. The isomer mixtures in the mixture of equal weights of the olefins of the formula XXIV (R ₇ = H), which are obtained in this way, are isolated by known processes, then the two are isolated.

Isomers of one another by known methods., For example. by chromatography. ,, separated. The less preferred.

009827/2053 EAD ORIGINAL

Isomerization of Formula XXIV then follows the same isomerization process subjected to obtain the preferred isomer. In this way, repeated isomerizations and Separation process essentially the entire amount doü less preferred Isoiaereii of the olefin of formula XXIY to the more preferred Isomers reformed.

The end compounds of formulas VII, VIII and IX, which are prepared by the process according to the invention, including the new end products of formulas X to XXIII in the form of the free acid, are obtained by neutralization with appropriate amounts of the corresponding inorganic or organic base, for example those listed above Cations and amines, converted into pharmacologically acceptable salts. These reactions are carried out according to a variety of known processes which are generally useful for the preparation of inorganic salts, ie, metal or ammonium salts, amic acid addition salts and quaternary nucleic acid salts ^ .fter. 1-s Jalzes produced from, in the case of the inorganic salts is os gewöhnlioh appropriate, the acid of formulas VII, VIII or IX in solving V tore it zn comprising a ilydroxycl, Cs-.i'bonat or Bic. arbonnt, d ⁿ .f: corresponds to the jeuHinoChten inorganic salt ^ in stücliiom · "." ::: isch-cr Mersgo. For example, by using sodium hydroxide, sodium carbonate or iron bicarbonate in this way, a solution of the n-trium salt is obtained The solid ano is obtained by evaporating the water or adding a water-miscible solvent of moderate polarity, for example a lower alkanol or a lower A1 organic salt.

To produce an amine salt, the acid is used in the formulas VII, VIII or IX dissolved in a suitable solvent of moderate or low polarity. Examples of the former are atlianol, acetone and ethyl acetate, examples of the latter are diethyl ether and benzene. Then a r; In-

009827/20 53

"" The stoehiometric amount of the amine corresponding to the desired cation is added to the solution. If the salt obtained does not precipitate, it is usually obtained in solid form by adding a miscible solvent of low polarity or by evaporation. If the amine is relatively volatile, any excess can easily be evaporated off. As a precaution, stoehionietric amounts of the less volatile amines are used. - ".". _:

Salts in which the ". Cation is a quaternary ammonium, are prepared by adding an acid of formula VII, VIII or IX "with a stoichiometric amount of the" corresponding quaternary ammonium hydroxide mixes in aqueous solution and then the water evaporated ".- .." ■ "■; ■■" / ""

As inclined in. Scheme A, - the ^ reaction participants XXIV, "'. XXV, XXVI and XXVII and _ furthermore the- corresponding ketals and tnao-I'iomers as well as the end products of the formulas, VIIj VIII and IX ₃ including PGE ₁ -, PGP ^ _f PGP _1ß , PGA ₁ and their lijomerc as well as the new .Compounds of the formulas X to XXIII each at least one ^: Äsymmetryzen1 »rum, and each of the compounds represented by these., Formulas exists in two optically active forms , namely the et-, and 1-Eorm. The above description of these compounds therefore includes the raceric di-? crm and the enantiomeric optically active d- "and 1-porms *;"

The optically active end products (d and l) of the formulas VII, 7111 and IX, including: PGE ₁ , ^1-3 GP ₁ ^, .PGP _{1 ß} and PGA ₁ , so / ie the. new connections of the formulas X to ΧΧΪΙΙ are (yes-; obtained by, one of the end products or one of the reaction parts ^ mcr of the formulas.XXIV, .XXV, XXVI, XXVII or ■. VII, which for their.production: used- If the end product VII, VIII or IX is a free acid, the dl form of the same is broken down into the d and 1 form by adding the free acid according to generally known methods an optically active base, for example brucine or strychnine, is reacted in order to obtain a mixture of two diasteroisomers ,

009827/2053

.. '· »' ···" "- BADORlQiNAL.

which according to well-known methods, "for example by fractional crystallization, can be separated to the individual to obtain diastereoisomeric salts. The optically active one Acid of formula VII, VIII or IX is then treated by treatment of the salt with an acid obtained by known methods. The olefin XXIV or glycol XXVI in the form of the free acid can also broken down into d- and 1-forms, then esterified and \ feiter -. " to the corresponding optically active form of the above, "end products VII, VIII or IX described above.

The olefin XXIV or glycol XXVI in the exo 'or endo form can also be converted into a ketal with an optically active 1,2-glycol, for example D - (-) - 2,3-butanediol, by converting the 'Glycol reacts with the compound of the formulas XXIV or XXVI in the presence of a strong acid, for example p-iouolsulfonic acid. The resulting ketal is a mixture of diastereoisomers, which is broken down into the d- and l-diastereoisomers, which are then both treated with an acid, e.g. Oxalic acid, can be hydrolyzed to the original keto compound (XXIV or XXVI), now in the optically active form with the diastereoisomers being separated from each other when the mixture of ketal diastereoisomers is used; then the optically active ketal of the compound of the formula VII is hydrolyzed with an acid, for example oxalic acid, to give the optically active compound of the formula VII. These reactions, in which optically active glycols and ketals are used for the decomposition, are well known; cf. formerly Ind., 'p. 1664 and J. Am. Chcm. Soc, Vol. 84, p. ₅ 2938 (1962).

09827/2053

- 46'- · : ■ /. '■: ■■■■■.' ■■ .- ■ '

Example 1 6-Exo- (1 '^' - erythro- and threo-dihydroxy-heptanyl) -2cc- (6 "-carboxyhexyl) -bicy'clo / 3.1 _f O_7 hexan-3-one (XXVI, R ₇ = H)

To 100 mg of 6-exo- (1 ^l -cis-heptenyl) -2 & .- (6 ^I '-carboxyhexyl) -bicyc-Io (3,1, oJhexan-3-one (XXIV), which under nitrogen to 0 ° a solution of 8 ml of dry formic acid (distilled over boric anhydride) to which TO / *.! 90% hydrogen peroxide and 65 mg of dry sodium bicarbonate had been added; all materials were prior to addition; with nitrogen flooded. After 30 minutes the ice bath is removed and the mixture is stirred for 1.5 hours lang.bei room temperature. the formic acid is removed in vacuo at 25 ⁰ C, then benzene was added and stripped liii vacuum to completely remove the formic acid. 10 ml of methanol and 2.5 ml of saturated sodium carbonate solution are added to the residue, the mixture is left to ^{stand at 5 °} C. overnight and then acidified to a pH of 4. The methanol is removed in vacuo, the solution is brought to pH -Value of 3 adjusted and extracted with ethyl acetate. The extracts are washed, dried, evaporated and chromatographed on 15 g of acid-washed silica gel. Elimination is carried out with 25, 35, 50, 75 and: 100% ethyl acetate-Skelly B mixtures (isomeric hexanes) and 1 and 10% ^: methanol-ethyl acetate mixtures. The initially eluted material (7 mg) has the same mobility in thin-layer chromatography as the starting material. 10 mg of a mixture of two materials, one of them with UV- * absorption, are now eluted. This is followed by 35 mg of a partially crystalline material which has an IR spectrum. and a V nuclear magnetic resonance spectrum has, in line with ':.. "; * the structural formula XXVI (R = H) The mass spectrum of this material shows a molecular ion (354) ^un ^ further 253 (cleavage between the hydroxyl groups of the glycol) as The then eluted material (with ethyl acetate) consists of 50 mg of a non-crystalline material which has IR and CMR spectra very similar to the preceding fraction

009827/2053

and shows a mass spectrum identical to this fraction. With both glycols the 6-endo-cyclopropy becomes hydrogen found in the nuclear magnetic resonance spectrum at 0.4-0.8 λ (multiplet), J = 3.5 and 7 cps; olefinic protons are not applicable.

Example 2 6-Exo ~ (erythro- and threo-J ', 2' -dilrydroxyhcptyl) -

2et »- (6 ^il -carbomethoxYhexyl) / 3,1 iPj hexan-3-one. (XXVI, R ₇ -CH ₃ ) '- "

To 1.90 g of 6-exo- (1'-cis ~ heptenyl) -2-K- (6 "-cai * bomethoxyhex3 ^ l) - / 3, IfO-"; --hexan-3-one (XXIV, R ₇ = CH ₅₎ at ⁰ ° C a mixture of? 0 ml of 98 $ formic acid, 650 mg sodium bicarbonate and 0.18 ml of 90 ^ sodium hydrogen peroxide are added, the on 0 ° 'cooled and purged with nitrogen. The solution is stirred for half an hour at ⁰ C, arrchließcnd it is turned off sum warming to room temperature for two hours. "The formic acid is removed in vacuo, and Ben sol is added. The latter is also removed in vacuo and the residue is extracted with ethyl acetate. The extract is washed with water, bicarbonate and saturated koch ales solution and dried with sodium sulfate. After evaporation, a residue consisting of formates of glycol is obtained. These are mixed with 50 ml of methanol and 10 ml of saturated. Large sodium bicarbonate solution hydrolyzed for two hours at 25 ° C. 40 ml Viayser are added, three methanol is added ir ¹ . The vacuum is removed and the aqueous suspension is acidified to pH 3 and extracted with ethyl acetate. The extract is washed with water and saturated sodium chloride solution, dried over sodium sulphite and evaporated. The residue is tan on 150 g of silica gel chromatography er ¹ : ur.d eluted with 750 ml of a 25, 35, 50 and 75% ethyl acetate-Skelly B mixture, and 150 ml fractions are collected. Fractions 12-15 (789 mg) consist of a; mixture of two isomeric glycols of the structure ZGIVI _; R ₁ -T = CH ^. ■ This material points to silica gel plates, 'those with the

0098 27/20 53

BATH

-48 - ■: ... "■ '

A. IX system developed a single spot with Rf = 0.62; a boric acid treated plate has two spots of almost equal intensity at Rf = 0.62 and 0.52. The upper spot corresponds to the less polar threo-glycol (see above). The nuclear magnetic resonance spectrum shows a 3-P: roton singlet at 3.67 ί (00H ₇ ); about 4 protons, multiplet, between 2.7 and 5> 6i -for ■ -. Carbinol and hydroxyl protons; 5 protons as a multiplet, 1.9 - 27 <f for protons that are adjacent to carbonyl groups, and one cyclopropyl proton at 0.6 α. Fractions 16 (6S5 mg) also show one spot. Silica gel plates, but two spots at Rf. 0.46 and 0.36 on boric acid treated plates; they consist of the more polar erythro and threoglycols (see below). The less polar isomer has the same mobility on plates treated with boric acid than the more polar threo isomer. Its IR and nuclear magnetic resonance spectrum are very similar to the * - those of the above fractions 12 - 15 ·. The mass spectrum shows 350 (K-1'8); - 332 (M-Hi ₂ O); 292 (350-58); 267 (M-101); 235 (267-32). . ::.

MiSEiSi ^ 6-Exo- (erythro-1 ', 2 ^ yypy (carböIαcthox3 ^Γ -hexyl) -bicyclo- ^ 3,1,0 / -hexan-3-one - (XXVI ₅ R ₇ = CH ₃ ) -

To a solution of 0.39 g of 6 - exo (cis-1 ^l heptenyl) -2i? '- (6 "-carbomethoxyheptyl) -bicyclo- / 3, 1,0 / hexane-one 3- ml in _8;; Pyrldin ve.rd.en 0.32 g Osmlumtetröxid ^ optionally After stirring at 25 ⁰ C v / hile I5 hours a solution of 1.0 g sodium bisulfite in 16 ml water and tO Pyrldin ml was added, and. Stirring is continued for 5 hours, the dark reading is diluted with water, extracted with chloroform, and the extracts are washed several times with water, dried and evaporated The residue is chromatographed on 50 g of silica gel Ethyl acetate eluted in Skellysolve 3. 0.0150 g des

009827/2053 _B AD ORlQINAL

less polar erythro isomers and 0.180.g of the more polar Erythro isomers obtained.

The less polar erythro isomer is crystalline (melting point: 70-71 ° C, from an ethyl acetate-Skelly B mixture); ν = 3460, 1730, 1715, 1250 ,. 1215, 1190, 1175, 1095, 1065, 1055 cm "¹; mass spectra at 368 (M ⁺ ), 350, 337, 319, 267, 250 and 235 mass units.

I ₁ calcd for C ₂₁ H ₅₆ O ₅ ; C = 68.44; H = 9.85 found: C = 68.46; H = 9.87

The more polyeric isomer is also crystalline] ν = 3430, 3340, 1735, 1710, 1.260, 1220, 1195, 1175, 1165, 1110, 1075, 1055, 1035 cm ", The mass spectrum is the same as for the weaker polar isomer. The melting point is 41 - 42.5 C after recrystallization from an ether ^: Skelly B mixture.

Anales6j_ found: C = · ■ 68.63; H = 9.79.

B.?J:ÄBiJiJ; 6-exo- (threo-1 ', 2 ^t -dihydroxyheptyl) -2c ^ - (6 "-carbomethoxyhexyl) - bicyclo £ 3.1, 0 / hexane ~ 3 ~ one (XXVI, R ₇ = OH ₇ )

In the same way as in Example 3 above, the trans isomer (500 mg) gives 180 mg of a less polar glycol and 110 g of a more polar glycol. The two compounds are not obtained in crystalline form, but their behavior in thin-layer chromatography corresponds to the two glycols obtained by hydroxylation of cis-XXIV (R ₇ = CH,) with performic acid (Rf = 0.62 and 0, 46 on boric acid-treated plates).

009827/2053

?.? ^ j 5 \ ■ --6-ExO- (erythro-1 ', 2'-dihydroxyheptyl:) -2ß ~ (6 "-carbomethoxyhexyl) -t) icyclo- ^ 5,1, OT -hexane-3 -on (XXVI, R ₇ = CH ₃ ). '·

In the same way as described above, 0.50 g of 6-exo- (1 ^I -cis-heptenyl) -2β- (6 "-carboxyhexyl) -bicyclo- [ 3.1", OJ -hexane 3-one (XXIV with 0.42 g of osmium tetroxide in 10 ml of pyridine. chromatography of the crude product are obtained 283 mg of the less polar erythro isomer XXVI ₅ R ₇ = CH _3, melting point of 42 ⁰ C (from ether-Skellysolve- B), Rf = 0.40 on silica gel, developed with ethyl acetate.

> ■ ■■■■ '■ ^:

Analysis calculated for C ₂₁ H ₃₆ O _n -; G = 68.44; H = 9.85 'Found: C = 68.21; H = 9.80. ■

The more polar erythro isomer consists of 148 mg of crystalline material, melting point: 58 - 59 ⁰ C (from ether), Rf = 0.19.

Analyze, found: C = 68.27; H = 9.37

These two glycols have almost the same IR and nuclear magnetic resonance spectra: ν = 3400, 1735, 1745, 1240, 1200, 1170, 1060, -735 cm ^-1 and 3.65 h, 3 protons as singlet (OCH ^); 4 protons as a multiplet, 3.4-2.9 · } (carbinol and hydroxy protons), 6-endo-cyclopropyl hydrogen at 0.48 ^; .

Example 6 6-ExO - (. Threo-1 ', 2 ^l -dihydroxyheptyl) -2ß- (6 ⁱ ' - ■

carbomethoxyhexyl) -bicyclo- / 3,1,0 / -hexan-3-one . "■■■■ (XXVI, R ₇ = CiV) '' ■■■ · ■"

Using the same procedure described above, 0.50 g of 6-exo- (1'-trans-heptenyl) -2β- (6 "-carboxyhexyl) -bicyclo- ^ 3,1, o] -he _/ xan-3- on (XXIV) hydroxylated with osmium tetroxide, and 219 mg of the weaker polar threo isomer and 129 g of the more polar threo isomer of structure XXVI, R ₇ = CH, are obtained. The weaker polar material is obtained from an Äblier Skellysolve-B -Mixture crystallized;

009827/2053

- 51 - ■

Melting point: 46 - 47 ⁰ C, Rf = 0.40 (Kieselsäuregelplatte, developed with ethyl acetate).

Analysis calculated for C ₀₁ H ₇ Jf-O, -: C = 68.44 H = 9.8-5 found: G = 68.31; H = 9.75

The more polar threo isomer is recrystallized from ether; Melting point: 77 ^{- '78 0} G, - Rf = 0.23,

Analysis found: C = 68.58; H = 10.11

Thin-layer chromatography of the animal glycols of structure XXVI, ß-side chain, R ₇ = CH ,, on silica gel plates which were sprayed with 10biger boric acid in methanol and dried for 30 minutes at 70 ° C, gives the following Rf fourth : weaker polar erythro isomer: 0.75; weaker polar threo isomer = 0.70; more polar threo-isomer = 0.60; and more polar erythreο isomer = 0.46.

The IR spectrum and KMR spectrum of the threo isoners are the Spectra of the above erythro-Ir. emers very similar.

Example 7 dl-prostaglandin LL-methyl ester and dl-15-isoprostaglandin E.-methyl ester

A. A solution of 509 mg of the more polar erythru number of XXVI, C * .- side chain, R ₇ = CH ,, in 9 ml of pyridine is ^{cooled to 0} ° C. and treated with 1.3 ml of methanesulfonyl chloride. " After one hour at ⁰ ° C., the ice bath is removed and the mixture is stirred for a further hour. It is now ^{cooled again to 0} ° C., ice is added, and the products are extracted with methylene chloride. The extract is diluted with cold Washed hydrochloric acid, dried and evaporated. This, ■ this crude pearly acid residue shows essentially a spot on thin-layer chromatography, shows strong IR absorption at 1740, 1350, 1175 and gto cm "- ■ and ^" excellent ion peaks in the mass spectrum at 532 (M-2GHaS © ₅ H), 301

009827/2053 ^ L

(332-32) mass units, the residue is rather unstable compared to the usual chromatography purification processes. It is dissolved in 27 ml of acetone, diluted with 13.5 ml of water, and the resultant solution is turned off for six hours at 25 ⁰ G. The «- mixture is concentrated in vacuo, extracted with methylene chloride, washed, dried and evaporated. Chromatography on 50 g of silica gel and washing with increasing proportions of ethyl acetate in Skellysolve-B gives the following materials: A. 75 mg of unknown, slightly polar products; B. 389 mg of a mixture of two glycol moiiomesylates; C. 27 mg ( ^{5.3 c} / °) R-isomer, dl-15-isoprostaglandin-E ^ -methyl ester; D. 34 mg '(6> 7v ^) crystalline dl-prostaglandin E. methyl ester (S-Isocier). . .-. "-. - -

Fraction B, ie the two monomes sy late, are marked as follows: Ions in the mass spectrum at 350 (M-CH ^ SÖ-, Η), 332 (350-18), 301 (323-31), 300 (332 -32), 319 (350-31), 31 3 (350-32) .. IR absorption at ν = 3500, 1745.1 340, 11 70, 920 cm ~ j and in the CMR spectrum a proton as a broad mu -I tiplett 5.0 - 4, .6 ^, 3 protons as a singlet at 3.7 <3 (OGH), one proton as a doublet of doublets 3.41: '»J - 7.5" and 3.5 Gps , a 3-proton singlet at 3.1 / (S-CH,) and 6-endo-cycloropropyl hydrogen as a multiplot at 0.85 - ..- 0.3

Traction C, ie dl-15-isoprostaglandin — S ₁ -methyl ester, has an IR spectrum and ÖIR spectrum- which are identical to those of the optically active 15 (R) -PGE ₁ -ZnGtIIyIGStCrS. The mass spectrum shows the molecular ion, 366, and other ions at 350, 332, and 297 vol units.

Fraction L is recrystallized from an ether-Skellysolvo-BG-emic "; melting point; 55-57 ° C. KMR spectrum, IR spectra, fc'Ov / ie the" / "obtained in the Bünnachicht chromatography are identical to that of natural PGE ₁ methyl ester. The mass spectrum shows peats at 368, 350, 55-2 "-xiride. 297 letting units; UY absorption at 278

/ 009827/2053 gADORIQiNÄL

(t = 26000, developed after adding a small amount 50 $ aqueous KOH to a sample in ethanol.

Analysis calculated for C ₂₁ H ₅₆ O ₁ -: C = 68.44; H = 9.85 found: 0 = 68.08; H = 9.92

B. The weaker polar erythro isomer of structure XXVI ^ x, side chain, R ₇ = CH ,, treated as above gives 5 ^ dl-PGE .. methyl ester and an equal amount of the 15-epimer. -

C. The more polar threo isomer of structure XXVI, € * · side chain, R ₇ = CH ₅ , also gives 55S ClI-PGE ₁ methyl ester and 5.5 f of the 15-epimer.

D. In the same way as described above, will the monomesylate gene obtained from the above solvolysis reactions converted to the bis-mesylate and solvolyzed to give a $ 5 yield of dl-PGE.-methyl ester and to obtain an equal amount of the 15-epimer.

SPIE l 8 dl-prostaglandin E

(Mg 329) To a dry sample of glycolic acid XXVI, <-? ', R ₇ = H, which consists .- side chain from the more polar erythro and threo-isomers, 20 ml of methylene chloride, 3.3 ml of trichloroethanol, 1.8 ml of pyridine and then 0.33 g of dicyclohexylcarbodiimide added. Subject to stirring for 2 hours at 25 ° C., the entire reaction mixture is poured onto a column of 100 g of silica gel. After eluting with 2 l of a 25775 ^ -igen ethyl acetate-Skelly-B mixture, 3OO mg of the desired ß, ß, ß-trichloroethyl ester of 6-E: to- (1 ', 2' -dihydroxyheptyl) -2cX- ( 6 "-carboxyhexyl) -bicyclo- / 3,1, θ] - hexan-3-one (XXVI, O ^ side chain) which is contaminated with some crystalline dicyclohexyurea. This material has a 2-propone singlet 4.76 ^ for the protons of the trichloroethyl group and is otherwise the same as the corresponding one

0 09827/20 5 3

1337908

Methyl ester XXVI, Ot side chain, R ₇ = GH ₃ .

The glycoltricliloroethyl ester obtained as above (300 mg) is ^{treated with 0.8 ml of methanesulfonyl chloride at 0} ° C. under nitrogen in 7.5 ml of pyridine. After 5 minutes at ⁰ ° C., the mixture is turned off in order to warm to room temperature, stirred for a total of two hours, cooled again in ice, and 5 ml of v / ater added. After 5 minutes stirring at 0 is added ethyl acetate, and the mixture is again washed twice with water, twice with 1N hydrochloric acid and then with bicarbonate and saturated brine, dried and evaporated ψ over sodium sulfate. The residue (348 mg), which was obtained by thin-layer chromatography, consists for the most part of a material that is less polar than the starting material, 8 ml of acetone and 2 ml of water are added and the resulting solution is stored at 5 C for 68 hours . Water is now added, the acetone is removed in vacuo, and the products are extracted with ethyl acetate. The extract is washed with bicarbonate and saturated sodium chloride solution, dried over sodium sulfate and evaporated> the raw weight is 315 mg. Do the chromatography on 50 g of silica gel and gradual elution with 1 liter of 25-1005 $ ethyl acetate-Skell7 / -B- and then mix with 5% methanol-ethyl acetate and get four tips.

Fraction No. 9-15 (88 mg) and No. 17-26 (117 mg) exist from monomesylates.

The IR spectra are very similar; OH (3500 cm " ¹ ); C = Q> (1745 cm" ¹ ), 1340, 1170, 920 * 800, 720 em " ¹ *

No. 32-40 (15 mg) moves a little faster in thin-layer chromatography than the methyl ester of 15-1So-PGE ₁ and undoubtedly consists of its corresponding trichloroethyl ester.

009827/2053

No. 43 - 47 (12 mg) moves a little faster in thin-layer chronatography than the methyl ester__ of PGE ₁ and shows two olefinic protons in the CMR spectrum, which are centered at 5.65 Λ; two protons of the triehloräthyestergruppe as a singlet at 4.76 </; otherwise there is agreement with the spectrum of the triehloroethyl ester of PGE ₁ .

According to the same one described above. In the process, 200 mg of the weaker polar erythro-threo pair of glycols with the structure XXVI ^ side chain, R ₇ = H) are converted into the trichloroethyl esters and bisaesylates and * solvolyzed to give 8.5 mg of the trichloroethyl ester of dl-PGE .. which is identical to that described above.

The above fractions 43-47 (12 mg) are dissolved in 1 relative 90% acetic acid and stirred with about 100 ng zinc dust at 25 ° C for two hours , at which point the thin layer chromatography shows that there is no ester lagged behind. Ethyl acetate v / ii ^ d to ^ e ^ just, the solution is decanted into a separating funnel, washed several times with water and then with saturated sodium chloride solution, dried on sodium sulfate and evaporated. The residue is a C at 3 g silica gel chromatography ex · f .. .nnri with 50 ml: mixture of respectively "50, 75 und.100 eluted y of ethyl acetate and Skellysolve-B and a mixture of 5 ^ methanol and ethyl acetate.. . fractions 7-9 are largely crystalline, move in the Dünnschi.chtchromatographie as FGE ₁ and melt after two Umkristallicationen from an ethylacetate-Skelly-B mixture at 113.5 to 115 ⁰ C; weight; 3.4 mg . ' The melting point with natural!:. FGE ₁ (melting point: 1.14 -115 ° C) is 109 - 1H ⁰ C The curve of the optical rotational dispersion shows no optical activity between 475 and 230 myu. (Accuracy 0.001 °), and the mass spectrum _{cit is identical to that of natural PGE 1.} The biological activity exceeds 50 5S of the natural PGE 1 in two systems.

009827/2053

193790a

The monomesylate fractions obtained in the previous experiment are again mesylated and solvolyzed as described above, and the dl-P & E ^ trichloroethyl ester fractions (approx. 6 Hg) are hydrolysed with zinc and acetic acid, in this way another 2 rag dl-PGE are obtained. . With a melting point of 113-115 ⁰ C, so that the total yield of pure dl-PGE,. 5 _{S is} 4 g or a total of 1.6% of the glycolic acid. . ^: .

Example 9 dl-8-isoprostaglandin B ^ methyl ester

A solution of Ö, 50 g of the more polar erythro-glycol of the structure: XXVI (O ^ side chain, R ₇ = CH ₅ , melting point: 5B- 59 ⁰ C) in 10 ml of pyridine is cooled to ^{0 0 C and mixed with 1} * 25 ecm methanesulfonyl chloride treated. The solution is stirred for one hour at 0 ° and then turned off to warm to room temperature over an additional hour. The mixture is then cooled, ice is added and the product is extracted with methylene chloride. The extracts are washed with cold 5% hydrochloric acid, dried and evaporated, 0.71 g of a brown oil remaining.

The other three Raeemates of structure XXVI (β-3 side chain, R = CH-) are converted into bismesylates in the same way. The migration speed of these products on silica gel plates which have been developed with a 50 follow Äthyläeetat-Cyclohex.aTi-Genisch is as follows: more polar erythro- and threo-bisiaesylates, Rf = 0.47; weaker polar erythro- and threo-bisciesylates, Rf = 0.57. All four give equivalent IR and KMR spectra; ν = 1745, 1360 »1180, 970, 915» 740 cm ""¹; a-proton multiplets transmitted to the mesyloxy group at about 4.8 and .4,3 (f (the proton of carbon atom), 5-proton Siiigulett at 5 »65 and 6-Pro-.tonen singlet" 3.1 <£ (OCH ₅ and SCH ₅ ) and a distorted triplet at 0.9 ^, corresponding to the terminal methyl group, which partially covers the e-endo-cyclopropyl hydrogen.

009827720B3 originally inspected

The bismesylate from the weaker polar threo-glycol crystallizes in the form of needles from the Ethylaeetat-Skellysolve-B-Gemiöch from °, melting point: 86 .- "87 ⁰ C.

Analysis calculated for C ₂₃ H ₄ O ₉ S ₂ : C = 52.65; H = 7.68; - 'S = 12.22.'. · ■■

found: C = 52.32; H = 7.74;

S = 12.21. -.

The crude bismesylate from the more polar erythro-Glyeol "is solvolyzed for five hours at 25 ⁰ C in 40 ml of acetone and 20 ml of water. After the usual workup, the crude products are chromatographed on 60 g of silica gel with 10-100 ethyl acetate ^ -igein eluted in cyclohexane The following materials were eluted in the order of their polarity: 94 mg of weaker polar monomesylate of glycol XXVI (β side chain, R ₇ = CH ₃ ) JB 341 g of an incompletely separated mixture of a more polar monomesylate of glycol XXVI (ß-3 side chain, R ₇ = CH ₃ ) and dl-15 ™ iso-8-ISO-PGE ₁ -inethyl ester; C. 50 mg (10 ^ yield) dl-8-iso-PGE ^ methyl ether, D »10 mg of dl-PGE.-methyl ester. They have the following characteristics.

A. v = 3400, 1745, 1175 and 920 cnT ¹ , ^ ?? ¹¹ 220 in / «.

- max f

(30007,

whereby a selected contamination by PGA ₁ -like compounds is indicated «

B. This mono-esylate, which (as was determined by thin-layer chromatography) contained impurities in the form of dl-1 5-iso-8-i3O-P & S ₁ -nethyl ether, is partially crystalline and is recrystallized from an acetone-Skellysolve-B mixture . Pure monomesylate is obtained; Melting point: ^{93-94 0} C, ν = 3510, 3450, 3030> 301Q ₅ 1745, 1540, 1205, 1175, 1170, 1030, 990, 920 and 810 cm " ¹ .. In the CMR spectrum shows a 3-proton -Singlet at 3 _? Q5o the presence of an S-CII, -group & n.

, «X ; : ,, .- äW. 009827/2053

Analysis calculated for C ₂₂ E ₅₉ O ₇ S: C = 59.17; H = 8.58; S = 7.18 found: G = 59.39; H = 8.77; S = 7.00

C. This material shows the same mobility (Rf = 0.40) on silica gel plates which had been developed with ethyl acetate- 'as authentic 8-iso-PGEj-methyl ester and, after treatment with a base, gives ^ x ² ^ ⁸ Εΐχ * (23400).

After recrystallization from an ether-Skellysolve B mixture, the melting point is 52 - 53 ° C, ν = 3400, 1740, 1240, 1200, 1175, 975 cm-. The nuclear magnetic resonance spectrum is the same as for authentic material and shows a broad multiplet: 5.0-5.8 for the olefinic G ₁ , C, ... protons. Significant ions at 350, 332 and 277 are present in the hate spectrum.

Analysis calculated for GP ₁ H ₇ ^ Of-: C = 68.44; H = 9.85 found: C = 67, SOj H = ■ 9.98

The bismesylate produced from the weaker polar threo-glycol XXVI (β-side chain, R ₇ = GH-) gives, after solvolysis, as described above, a 5.4% yield of dl-8-iso-PGE.-methyl ester. The more polar threo-glycol XXYI - (ß-side chain, R ₇ = CH-) gives a yield of 11%, from the less polar erythro-glyeol XXYI a yield of 8.0 %. Solvolysis of the slightly polar erythro-bismesylate in a tetrahydrofuran-water mixture (2: 1) gives, in contrast to the above, 10 $ dl-8-iso-PGE ₁ -methyl ester. In both cases it also forms 1-2 $ dl-PGE ₁ methyl ester. After solvolysis, one of the monomesylates in crystalline form (melting point: 85-87 ° C.) from an acetone-Skellysolve-B mixture is obtained from the more polar threo-bismesylate. . '■

Analysis calculated for C ₂₂ H ₅₈ O ₇ S: G = 59.17, H = 8.58

found: G = 58.95ί H = 8.72; S = 7.01., -

009827720S3

BATH ORIGINAL

■ -. 59 -

Example 10 8-Iso-irostaglanöin E ₁

To 1.2 g of δ-Εχο- (1 '-cis-heptenyl) -2ß- (6 "-carbcxyhexyl) -bicyclo - / * 3,1, Öj -hexan-3-one (XXIV) is a cold one under nitrogen (0 °) solution of 325 mg sodium bicarbonate and 0.1 ml 90% hydrogen peroxide in 25 ral 98% formic acid. After stirring at 0 ° for 0.5 hours, the ice bath is removed and stirring is continued for a further 1 The aric acid is removed in vacuo, 25 ml of benzene are added and also removed in vacuo, the residue is extracted with ethyl acetate, the extract is washed with water and saturated brine, dried with sodium sulfate and evaporated the residue are added 45 ml of methanol and 15 ml of saturated aqueous llatriumbicarbonatlösung ·. the mixture is stirred for two hours at 25 ⁰ C, the methanol concentrated in vacuo to remove and acidified to a pH of 2-3. the products are with ethylacetate extracted; the 'extract is washed with water, dried and evaporated to give 1.38 g of a mixture of epimeric glycolic acids (3XVI, fl-side chain, & pH). This "is esterified in 140 ml Methylencnlorid with 23 ml of trichloroethanol, 12.6 ml of pyridine and 2.31 € Dicyclohexylcarbodiinid at 25 ⁰ C for 2 hours. Then 5 nil v / ater was added and after fünfminütigei 'stirring, the mixture is treated with Washed 1N hydrochloric acid and liatriuüibicarbonatlösung, ßetrnolmpt and

■ evaporated. The residue is dissolved in benzene, filtered to remove cyclohexylurea and chromatographed on 150 g of silica gel. Eluting with 20-100 $ ethyl acetate in Skellysolve-B gives main ir alctions which form the epiEate glycols of the structure SXYI ( ß-Seitenlcette, a ^ OHoCOls) correspond. The ßclLwäoher polar material (700 .log) shows a spot in the DüimseMehtehroiaatagraphie (silica gel, 50 ^ iges Atftyladetat ”-cyclohexane Gemlscli), the starter polar (800 mg) Seigt two in the same system

ί, sehff close to beieinanaerliegensiö spots. The Ef values of these * 2ridxLoräthylester are similar, but slightly increased compared to

ORIGINAL INSPECTED

over that of the methyl esters described above.

To 66? mg of the weaker polar glycol XXYI (β-side chain, R ₇ = CH ₂ CCl,) are added 16.5 ml of pyridine and then at ⁰ ° C. 1. 67 ml of methanesulfonyl chloride. The mixture is stirred at 0 ° for ten minutes and then a further 1 3/4 hours while removing the ice bath. Now it is cooled again to 0 °, 15 liters of water are added, and the mixture is extracted with ethyl acetate. The extract is washed several times with cold diluted hydrochloric acid and aqueous liatric bicarbonate solution, dried and evaporated. The raw _. Bismesyiat is stirred in 53 ...- 2il acetone and 26 ml water under nitrogen at 25 ⁰ C üb.er night. It is / trated in vacuo conc s, extracted with ethyl acetate, the "extract is washed, dried and evaporated. The residue operatively on 75 g of silica gel ehromatographiert and eluted with increasing amounts Äthy'lacetat-in Skellysolve-B. After two main fractions, namely of 276 mg and 103 mg of unreacted glycol monofiesylate, 34 mg of 8-isoprostaglandin-E ^ -trichloroethyl ester with two olefinic protons: between 5.0 and 6, O ^ "(H ₁ . ^ -at- 5, 28 £, S- 15 and 9.5 Gps, and H ₁ ■. at 5, 1d, ■ J- 15 and 7 Gps), two protons ■ of the triehloroethyl group as a singlet, 4.8 / f , and two cooking

. binol protons as multiples between 3.9 and 4.5.-.- Co This is followed by 12 mg of the riclilora-ethyl ester of prostaglandin-B ^ its KiiR spectra. is identical to that of the iProstaglandin-E ^^ trichlprathylester described above,

The above-erlialtenen 34 & g 8 ~ isoprostaglandin _E-1 - * - tricliloräthylester v / ground in 1 Luel 90 ^ i; ger dissolved Essigsätire and 2wel hours nit 100 mg ZiiikBtaub stirred. The mixture is: now with ethyl acetate; diluted, washed several times with water, drunk kne-b ttnd evaporated. Ώ & χ residue is eluted on-5 g ⁱ 'Silicar-C'C4 "(MaHinckf odt), with acid; washed silica gel, cliromatograph and with 50 ~ TÖÖ ic Ethyl-αοβ ^ Βφ-3ίς6ΐΐ73θ1νβ-Β"& βΏΧ3ϋ ^. The fractions (15 mg), which in their thin-layer chromatography mobility

the 8-isoprostaglandin E .., A IX system correspond, are pooled and-Skellysolve-B mixture Ätnylacetat crystallized from a (melting point: 101 ~ 102 ⁰ C). _9. This material shows no optical rotation 475-230 m / c, and the nuclear magnetic resonance and mass spectra are identical to those of the "natural" isomer.

Analysis. Calculated for C ₂₀ H ₅₄ O ₅ : C = 67.76; H = 9.76 found: C = 67.56; H = 9.60

Example__1_1 dl-prostaglandin-A ^ - and dl-prostaglandin-B.-methyl ester

A mixture (150 mg) of the weaker polar er3rth.r0- and threo-glycols XXVI (oC side chain, R ₇ = CH ₅ ) is ^{treated in x} 4 ml of pyridine with 0.4 ml of methanesulfonyl chloride at 0 ° and then for two hours turned off uni to warm to room temperature. Ice is now added, the product is extracted with ethyl acetate, the extract is washed with cold, dilute hydrochloric acid and sodium bicarbonate solution, dried and evaporated. The crude bismesylate turns yellow in 15 ml of acetone, to which 2 ml of water and 4 ml of saturated sodium carbonate solution are added, and the resulting mixture is refluxed for four hours under nitrogen. After acidification, the mixture is extracted with ethyl acetate Extracts are washed, dried and evaporated. The crude residue is briefly treated with excess ethereal diazomethane and chromatographed on 20 g of silica gel. Eluting with increasing amounts of ethyl acetate-containing cyclohexane gives a total of 71 Bg (50% yield) of a mixture of dl-PGA.- and dl-PGB.-methyl esters. The first fractions of the main peak (14 mg) consist of pure dl-PGA ₄ esters, Λ ^Α ΐ °! Ί 221 _m / _C (i1 000; the mobility in various thin-layer chromatography systems is identical to that of the natural material 'Remaining material of the Ilaupt peak (56 ng .; Λ max Si 9 (7530); "278 (1015O) J

009827/20

is a mixture of 65% (H-PGA ₁ -methyl ester and 35% (H-PGB ₁ Methylester.

Following the procedure of Example 11, but using a mixture of the weaker polar erythro- and threo-glycols XXYI (β-side chain, R ₇ = CB ₅ ) as starting material, dl-8-PGA ^ methyl ester and ^ 1 are obtained -8-PGB ₁ methyl ester.

Example 12 6-Exo- (1, 2'-dihydroxyheptenyl) -2β- (6 "-carbomethoxyhexyl) -bicyclo- £ 3j1 _f oJ-hexan-3-one (XXVI, OC side chain, E ₇ = GH ₃ )

w A mixture of 4.73 mg of crude 6-3xo- (1 ', 2' ~ oxidoheitanyl) -2ß- (6 ^{! i} -carbomethoxyheptyl) -bicyclo- / 3,1, OJ-hexan-3-one (XXV, ß-side chain), which had been prepared from 5 »0 g of ice, trans-olefin, 9.5 ml of formic acid and 13.5 ml of methylene chloride is left to stand for 15 minutes at room temperature. The mixture now turns to a dark oil under reduced pressure. evaporated, which is dissolved in 250 ml of methanol ;. 102 ml of saturated aqueous sodium bicarbonate solution are added, the mixture is purged of oxygen by a stream of nitrogen and stirred for 4.5 hours at room temperature. After concentration to 1/4 of the volume, the mixture is extracted four times with methylene chloride, the combined extracts are washed with water, dried and evaporated. Gross weight: 4.70 g. The oily material is chromatographed over 500 g of silica gel as follows (SSB = Skellysolve B);

2 _S 5 1. 25 ^ AtOAc in SSB ^
-..- ■: t / 5 1. 50% ÄtOAc in SSB? ¹ ^ ⁷⁸ € ^ ^Thrown

3.5 1. 50 ^ AtOAc in SSB / '

> 1 _s 24g Glycol 1 Rf, 10D ^ AtOAc :,

1.0 1.75 ^ AtOAc in SSB) ^ V _; ;. ⁰ »4 °

4.0 1.75 $ AtOAc in SSB /

^ 0.75 S Glycol 2 Rf, .100 ^ AtOAc, 0.5 1. 100? Έ ÄtOAc. J ■■: -. 0.23 x 5.0 1. iOO ^ ÄtO ^ ci "0 ^ 62 g Glycol 3 Rf, 100 ^ AtOAc,

■■ &'& t ^: ---- /; ---- ₀₁₉ .,;.

Glycol 1 is a mixture of weaker polar erythrocytes and weaker polar threo shape; Glycol 2 is the more polar ^ threo form, and glycol 3 is the more polar erythro form,

Following the procedure of Example 12, but using 6-exo- (1. ' _f Z ' -osidaheptanyli - ^ - Co ^ carbomethOxyheptyl) -bicyclo- [3, 1, oj -hexan-3-one (XXV, ^ Side chain 11 e) as starting material, one obtains the properties corresponding to the above β-side chain glycols ^; - ~ side chain glycols of the formula

Example 13 .8-IsO-PGE ₁ from

A Ijosung 1,00.'g of PGE ₁ and * 5 g Käliumacetat in 100 ml of 95 ii # ^e "- ethanol is abgösteilt at room temperature under nitrogen for 6 days and then by evaporation under reduced pressure to about 1/3 of its volume . concentrated the concentrate e mixture is diluted with 75 ml of cold V / ater and dilute Sal2s; iure is added until the mixture has a PLL ^ ^r ert \ has erreiGht 3 the acidified mixture is extraliiert twice with ethylacetate.. , then saturated with sodium chloride and extracted again with ethyl acetate. The ethyl acetate extracts are combined, washed with saturated aqueous sodium chloride solution, dried over sodium sulfate "-, -under reduced pressure and then evaporated under a stream of nitrogen to remove the ethyl acetate from the residue dried, analysis by thin layer chromatography shows that the residue is a mixture of PGE ₁ , 8-iso-PCtE ₁ and PGA _1. Each recrystallization of this residue: from a em mixture of acetone and. Skellyöolve B is obtained. Öj43g crystalline PGE ₁ , which was determined by analysis to be pure. The mother liquors from the crystallization are chromatographed on 50 g silica gel (GG-4) and developed with a mixture of ethyl acetate;

0ÖS827 / 20S3

, - "■■ ■, - ■ ■■■■■■ - 64 -. ' ■;; '■■;.;.

Fractions 1-10 40 $ ethyl acetate ~ 60 % cyclohexane

. . ■ ■■ ■ - ■ '- /' ■ ■ .- ■ "■ '■■ ■"' ■ '"■" - ■'. ·. "■ ---

Fractions 11-20 * 50 $ ethyl acetate -50 $ cyclohexane Fractions 21-25 $ 60 ethyl acetate - $ 40 cyclohexane Tractions 26-30 $ 100 Ethyl Acetate '

The fractions are evaporated, and the residues are analyzed by layer chromatography (silica gel, developed according to the Bush A-IX system), fractions 23 - 25 (55 mg) contain 8-iso-PGE ^, and after crystallization from an acetone Skellysolve-BG-emic gives 46 mg of 8-1SO-PGE ₁ , the melting point of which is 72-75 °. Fractions: 27-30 result in 41 mg PGE after crystallization. . Fractions 5-9 (399 mg) consist "as demonstrated by thin-layer chromatographic analysis, of PGA ₁ .

Example 14 PGE ₁ from 8 ₁

2.2 g of potassium acetate are added to a solution of 100 mg of S-iso-PGE ^ in 50 ml of 95% ethanol. After the potassium acetate has dissolved, the mixture is turned off at Raiimt.Rmpm-rH - nx · under nitrogen for 92 hours; then it is concentrated under reduced pressure to give a residue remains, w'asser v ^"r ird added to this, then v / ith 1N hydrochloric acid dem'Gemisch added to bring the pH to about 3, and dc.s Mixture is extracted with ethyl acetate. The Äthylac etat extract vird "V / ater, then washed with saturated-aqueous sodium chloride, dried over sodium triumsulfat uad under reduced pressure to ->: -.-'.- evaporated; isan a pale yellow receives." Residue ^1, the PGE ₁ contains, as demonstrated by thin-layer chromatography. This residue is crystallized twice from an ethyl acetate Skellysolve B gel and obtained from the second crystallization. 57 PGE ₁ mg with a melting point of 11-2 - 114 ⁰ C. The combined mother liquors from both: crystallizations are evaporated under reduced pressure,

the residue is treated with potassium acetate in ethanol, and, as described above, ■ worked up, with one more 12 mg crystalline PGE. receives. "·

Example 15 6-ExO- (1 '-cis-heptenyl) ~ 2o6- (6 "-carboxyhexyl) · -

bicyclo / 3,1,0 / -hexan-3-one '(XXIV)

A solution of 440 mg of 6-exo- (cis-1 ^! -Heptenyl) -2 ^ - (6 ^iI -carbO-methoxyhexyl) -bicyclo- / 3,1, oj-hexan-3-one in 48 ml of isopropyl alcohol is in chilled in an ice bath. A solution of 430 mg of sodium borohydride in 4.8 ml of water is added with stirring. The mixture is stirred for 21/2 hours, the ice gradually melting and the temperature rising to about room temperature. Now 2 ml of acetone and then 2.4 ml of acetic acid in 24 ml of water are added. The organic solvent is removed in vacuo and the residue is extracted with ethyl acetate; the extract is washed with bicarbonate solution and saturated sodium chloride solution, dried over sodium sulfate and evaporated. The crude mixture of 3-alcohols is dissolved in 16 ml of methanol and in 8 ml NatriumhydroxydlÖ-sung v / ground ziigegeben The mixture is stirred for two hours at 25 ⁰ O under nitrogen.. then wird'die clear solution by the addition of 5 ml water and 12 ml of acidified in hydrochloric acid The methanol v / ill in vacuo removed, and the product is extracted with ethyl acetate, the extracts are washed with water and saturated brine, dried with sodium sulfate and evaporated to give the residue a structure of 6-exo- (1 '-heptenylJ - ^ - ib ^ -carboxyhexylJ -bicyclo- β , 1/0 / - .. nexan-3-ol corresponding KMR ~ spectrum u

This crude product is dissolved in 100 ml of acetone, cooled to 0 ° and treated with 1 ml of Jones reagent for 10 minutes. so 4 ml of isopropyl alcohol and then 40 ml of water added, and the acetone is removed in vacuo. The product It is extracted with ethyl acetate, the extract is washed with 1N hydrochloric acid and saturated salt solution, with sodium sulfate

008827/2053

- 66 - ■ -.,: ■ ^: ■■ '■■ ". ■■■" / ■' ■. i ^:

dried and evaporated. The crude product is chromatographed on 50 g of Mallinckrodt-Siliear CC-4 and eluted with 5, 7 1/2, 10, 15, 25 and 50 μl of ethyl acetate in Skelly-B. Fractions 9-12 contain 303 mg of 6-exo- (1'-cis-heptenyl) -2 ^ .- (6 "-c-irboxyhexyl) -bicyclo- £ 3.1.0 / - hexan-3-one (XXIV). Ν = 2500 - 3500 (GOOH), 1745, 1720, 1040, 845, 725 cm " ¹ .

In the same way as above, 440 mg of 6-exo- (1 ^l -cis-heptenyl) -2β- (6 "- carbomethoxyhexyl) -bicyclo- / 3,1,0 | - hexan-3-one are reduced with sodium borohydride , saponified and re-oxidized. Chromatograph the crude product such as. carried out above, gives 274 mg of pure 6-exo- (i '-' ■ ciG-heptenyl) -2ß- (6 ⁱⁱ -carboxyhexyl) -bicyclo / 7-3,1, Oj-hexan-3-one "(XXIV in the fractions 12 - Η, whose KHR and IR spectra agree with the above formula.

Na.cn the procedure of Example 15, except that hexane-3-one is used as starting material 6-endo- (1 'cis-heptenylJ aei-Co ^ -carboiiiethoxyhexyl) bicyclo-r 3,1 _f 0 {, 6-endo- (1 ^f -cis-heptenyl) -2o (- (6 "-carboxyhexyl) bicyclo - [ 3,1,0f-hexan-3-one | when using 6-endo- (1 '-cis-heptenyl) 2ß- (6. ⁱ ' -carbomethoxyhexyl) -bicyclo - ^, 1, 0 ^ | -hexan-3-one as starting material, 6-endo- (i ^l -cis-heptenyl) -2ß is obtained - (6 "-carboxyhexyl) -bicycro- / 3,1, O ^ -hexan-3-one.

Example_16 8

A solution of 100 mg of 8-iso-PGE. methyl ester in 5 ml of isopropanol is cooled in an ice bath and stirred; 50 mg of sodium borohydride in 1 ml of water are added at once * After addition of the borohydride or 2 _r 5 hours stirred, while the ice bath melts. At this point in time, analysis by thin layer chromatography (silica gel, developed three times with ethyl acetate) of part of the reaction mixture shows that the starting material is essentially no longer present. Acetone is added and the mixture is

Stirred for 15 minutes; then it is diluted with Aqueous acetic acid is neutralized and, under reduced pressure, concentrated until most of the isopropanol away. The remaining mixture is extracted with ethyl acetate, the ethyl acetate extract is over sodium sulfate dried and reduced under reduced pressure to give an almost colorless residue, see below. This residue v / ird dissolved in 40 ^ igem ethyl acetate in cyclohexane, chromatographed over 10 g of silica and with 10 ml amounts of the eluted solvents listed below:

'~ Frjik_tipnen Solution-mi11el Eluted amount, mg

1-5 AÖfi ethyl acetate

Cyclohexane

6-10 6650 ethyl acetate - -

34 / ö cyclohexane

11 100 ^ ethyl acetate 2

12 "" '1

14 "-ι. - 4 - ■

15 " ^l! 42

-If ' 5f- methanol - 95> - ethyl-e-

did .,. ■ '58 (crystalline)

"17 ■ ■. -. ·" ■ '»" 12

18 "^i;'^;" 7

19 ^{i (} "'-' 4

20 """^{>:; i} 2

. . Oil 'ii 11 i il -j

22 ¹ V """O

The fractions λ ^ Ι, 18, 19 and 20 v; are combined and, as mentioned above, pm = ut oln * oijiat-r> ^ i-ap} iiert. Corresponding fractions of the first and other öhx'oiHatograr.Vtrie are combined, se that a total of 61 mg of partially crystalline material (the more polar of the two products), 13 mg - a mixture (according to dtiiinschichtchroi; iatographic analysis ^ mainly the starch "polar product) , and 15 mg of one

009827/2053

weaker polar product, i.e. / 0-iso-PGF ₁ O can be obtained.

The more polar fraction (8-iso-PGF.-.) Is crystallized from a mixture of ether and Skellysolve B (mixed hexanes), and 8 mg of 8-iso-PGF .. ^ are obtained in the form of waxy crystals with a melting point at 60 - 61 ⁰ O ^.:

Example-.1-7. Equilibrium between 6 — Exo- (cis-1 '-lieptenyl) -2t5i- (6 "-carboxy-methoxyhexyl) -bicyclo - £ j, 1, Oj-hexan-3-one and its 2-isomers

1.2 g of 6-exo- (cis-1 .'- heptenyl) -2i7 ⁽ '- (6 "-carbomethoxyhexyl) -bicjrclo- / 3,1,07-hexan-3-one are dissolved in 100 ml of dry dimethoxyethane then, potassium tert-butoxide (400'mg) is added, and the mixture is kept under nitrogen for one hour at 25 ⁰ C. v / ith hydrochloric acid (n -3) is added in sufficient quantity to the Kaliurn-tert The mixture is diluted with 500 ml of water and then extracted three times with 100 ml amounts of ethyl acetate. The ethyl acetate extracts are evaporated to give a residue which is chromatographed on silica gel (eluting with a 5 / o ethyl acetate -Skellysolve-B mixture). After evaporation of the eluates, 380 ng of the starting material (*% _) and 700 mg of the corresponding β-isomer are obtained.

Following the procedure above, but using the β-isomers as the starting material, the same results were obtained.

009827/2053 BADOR.G.NAL

Example 18 ö-Carbäthöxy-bicyclo-j ^ ji, 0? -Hexan-3-ol and ö-Garbätlioxy-bicyclo- ^, 1, Oj-hexan-2-ol

A solution of 96.46 g of δ-carbethoxy-bicyclo- / 3.1.0J -hexane in 500 ecm of dry ether is stirred under nitrogen; about half of 266 ml of 1.0 M borohydride in ether is added dropwise at room temperature The reaction mixture is ^{cooled to 0 0} O and the remaining borohydride solution is added. The addition of the borohydride solution takes about 45 minutes. The reaction mixture is now stirred at room temperature for 45 minutes, and then the solvent is removed by evaporation under reduced pressure. The residue is dissolved in 500 ml of ether and ^{cooled to 0} ° C. using an ice-methanol bath; 150 μl of 3% aqueous sodium hydroxide solution are now added over the course of 10-15 minutes, the temperature being kept below 5 ^° C., then followed the addition of 80 ml of 30 ^ ethyl hydrogen peroxide for 15 minutes, the temperature being kept below 1O ⁰ G. the mixture is then stirred for 35 minutes at room temperature, and the layers are separated. D The aqueous layer is extracted twice with ether and three times with ethyl acetate. The organic solutions are combined and washed with saturated aqueous sodium chloride, dried over magnesium sulfate, filtered and evaporated under reduced pressure. 89 g of a residue are obtained, which consists mainly of a mixture of 6-uarbäthoxy-bicyclo- / 3,1, Oj -hexan-3-ol and ö-carbethoxy-bicyclo- ^, 1, oJ-hexan-S-ol the 3-01. .

Example 19 6-Carbethoxy-bicyclo-f3,1, 0 / -hexan-3-ol.-tetrapyranyl ether and 6-carbethoxy-bicyclo- /; 3.1, 0 / -hexan-2-ol-te brahydropyranyl ether ^:

A mixture of 88.0 g of a mixture of 6-carbethoxy - bicyclo ~ / 3,1, o7-hexan-3-ol and 6-carbethoxy-bicyel. O - / 3> 1 ♦ 07-hexan-3-0I and 88 ml of dihydropyran is cooled to 0, and 40 drops

000827/205

■■ -. "■ '. - ■■.-_ ■'" ..- ■■■ - -

Phosphorus oxychloride are added. The mixture is stirred for 2 hours at ⁰ ° C. and then for about 18 hours at room temperature. The mixture is then diluted with methylene chloride and washed with cold aqueous saturated sodium bicarbonate solution. The layers are separated and the aqueous layer is extracted three times with methylene chloride Combine the organic layers and wash with water (back-extract the washings), dry over sodium sulfate, and evaporate under reduced pressure to leave a residue. This residue is distilled under reduced pressure to give 18 g of a 40-boiling forerun ⁰ C between 1.3 and 0.4 mm Hg ; 75.3 g of a mixture of 6-Garbäthoxy-bicyclo- / 3,1, C-hexane-3-oil-tetrahydropyranyl ether and S-Garbäthoxy-bicyclö were then obtained -l ^ ji, Oj-hexan-2-ol-tetrahydropyranyl ether with boiling point at 98-131 ⁰ C within a pressure range of 0.3-1.0 mm Hg.

Example 20 PGE ₁ from PGB ₁ methyl ester

A. Production of enzymes.

A medium is prepared which bealent from 2% corn steep liquor (a mixture of equal parts of Öerelose and glucose) in tap water. T> i «ötnj ff ^ diüm v / ill by adding hydrochloric acid to a pH of 4.5 ¥ ert geDiaokt, and \ f> methyl wijcd added. Animal 5ÖÖ-ml flasks each containing 100 ml of the above medium are resihae with Oladosporum (Gl-I1, ITCG 1f 274) and vaccinated (about-28 ⁰ C) geschüttert for four days at room temperature on a shaker. The culture is then placed in a 40 ml centrifuge tube and centrifuged at about 2000 rpm in a clinical centrifuge. The liquid is decanted from the centrifuge tubes and the cells are washed with cold water. The washed cells from two centrifuge tubes are placed in 50 ml of an ice-cold 0.05, M (pH = 7.0) phosphate buffer solution

■ '■'. BAD ORIGINAL .-. ■■ - ■■.; ■■

3 '' ■: ■ ^; "

1337908

suspended and placed in an ice-cooled Waring mixing beaker. Glass beads are added and the suspended ones Cells are agitated in the mixer for 15 minutes. The received Chopped cell suspension is run in a clinical centrifuge at approximately 2,000 rpm for 15 minutes centrifuged at room temperature, then the supernatant is collected. This liquid contains cladosporium resinae acylase and can be used directly for hydrolysis of PGE ^ -alkyl esters used or, preferably frozen, stored until needed.

B. Esterase hydrolysis of PGE .. -me t hy le st er

10 ml of the supernatant liquid containing Cladosporium resinae acylase prepared by the procedure of Part A in this example and 50 mg of PGE, methyl ester are shaken at room temperature under nitrogen for about 19 hours, then 70 ml of acetone are added and the mixture is filtered. Thin-layer chromatographic analysis (silica gel plate, developed with CHCl,: HOAc: CH, OH, 90: 5: 5) showed that both the filtrate and the insoluble residue obtained on filtration contain _{PGE 1.} The filtrate is evaporated under reduced pressure, and _{40-50 mg of a pale yellow oil containing PGE 1} are obtained, which is combined with the insoluble residue and chromatographed over 10 g of acid-washed silica gel (Silic ARCC->, Mallinckrodt). Elution is carried out with a hexane mixture (Skellysolve 3), which contains increasing amounts of ethyl acetate. The 50 ml fractions listed below were collected:

009827/2053

Solvent fraction

1 Skellysolve B.

2 40 ml Skellysolve B-10 ml ethyl acetate 5 "■« ■ "30" ^{! L} 20 ""

4 25 "" 25 "ti -.

5 20 «" 30 ""

6 10 ".." - 40 »» γ JH Κ 45 ""

8- ■■ ':.' Ethyl acetate '

1G "

- .-- 12 100 ml of ethyl acetate

Fractions 6 to 12 are combined and crystallized from a mixture of icetone and hexanes (Skellysolve B). The crystals are filtered off, washed with ether and dried; 30 mg of crystalline PGE ^ are obtained with a melting point of 115-116 ⁰ O and a mixed ^{melting point of 114-114.5 0} G. The curve of the "optical rotation is identical to that of the authentic PGE ₁ .

_fc C. Ssterase hydrolysis of dl-PGE ₁ -methyleBter

12 al of the supernatant containing Cladosporium resinae acylase, prepared as described in Part A of this example, and about. Maintained in the frozen state for 3 weeks are _{mixed with 129 mg of 1-PGE 1} -me thy read and shaken at room temperature under nitrogen for about 20 hours] then the mixture is diluted with 100 ml of acetone, filtered and reduced under reduced pressure Evaporated pressure to give a residue, the dl-PGE. contains. The residue is chromatographed on Silic AR CO-4 (Mallinck-rodt).

The column is filled with mixtures of Skellysolve B and ethyl

009827/2053

- 73 -
acetate as well as ethyl acetate and methanol eluted as follows:

fraction I. V V 50 ml solvent B. + 5 ml of ethyl acetate J 45 ml Skellysolve ί) 10 "». 1 40 ml Il Il VJI 35 ml Il 11th 20 » ^{! I} 30th ml Il Il 25th 25th Il 30th 2 20th 35 3 VJl 35 4th VJl 40 5 10 40 6th 10 45 7th VJl 45 8th VJI 9 50 ml 10 Ethyl acetate 1% methanol 11th 50 ml Ii ■ + Vk " 12th 1 Il H Ethyl acetate + Ho '» 13th 1 ti Il Il + ' 2 ° ß> » 14th ti ''. ti + % " VJl 00 Il il + 4 ^c / o ' ^: 16 00 il il ₊ 17th il

The fractions are evaporated and then by thin-layer chromatography analyzed. · Fractions 7-9 contain djl-PGEV-methyl ester (Total weights 66 mg), fractions 10 and 11 contain a mixture of d, l-PGE. and d, l-PGE ^ methyl ester (Total weight; 20 mg), and fractions 12-16 contain (Total weight: 33 mg).

Example 21 djl-i-iso-PGE ^ methyl ester from the endo series

A solution of 47 mg of 6-endo- (1 ', 2' -dihydroxyheptyl) -2J3- (6 "-carbomethoxyhexyl) - 'bicyclo- / 3 _> 1, o | -hexan-3-one in 2 ml

009827/20 53

dry pyridine is cooled and taken in an ice bath Stirred with nitrogen, 0.3 ml of methanesulfonyl chloride are added added, and the mixture is for 2 1/2 hours in the stirred melting ice bath. The mixture is then cooled in a fresh ice bath and diluted with 10 ml of cold water and stirred for 10 minutes. Finally, it is poured into a separatory funnel containing ice and with three 25 ml volumes cold ethyl acetate extracted. The ethyl acetate extracts are combined, with 15 ml of cold water, 15 ml of cold 10biger Sulfuric acid, 15 ml of cold aqueous 100% sodium carbonate

| washed with sodium solution and two 15 ml quantities of cold water; then it is dried over sodium sulfate and evaporated under reduced pressure to give a residue which is the bis-mesylate of 6-endo- (1 ', 2 ¹ -dihydroxyheptyl) -2β- (6 "-carbomethoxyhexyl) -bicyclo- ^ 3,1 This residue is dissolved in 2 ml of acetone plus 1 ml l / water and left at room temperature for about 18 hours, then the mixture is evaporated under reduced pressure, 10 ml of water are added and the mixture is extracted with three 20 ml quantities of ethyl acetate. The ethyl acetate extracts are combined and washed with 10 ml of saturated aqueous sodium bicarbonate, dried over lithium sulfate and evaporated under reduced pressure to leave a residue.

f which, as demonstrated by thin layer chromatography analysis (silica gel plate, developed with ethyl acetate), contains d, l-3-iso-PGE.-methyl ester.

Example 22 PGE ₁ and 15-PGE ^ from endo series

A solution of 0.115 g of 6-endo- (1 ', 2 ^l -diIiydroxyheptyl) -2rA .- (^ "- earbomethoxyhexylO-bicyclo-jfS, 1,0j-hexan-3-one in 4 ml of dry pyridine is placed in an ice bath cooled and stirred under nitrogen while adding 0.6 ml of methanesulfonyl chloride. Stirring is continued and the ice bath melts for about 2 1/2 hours. The reaction mixture is then cooled in a fresh ice bath with 10 ml of an ice water Mixture diluted and another 10 minutes

touched. The mixture is poured into a separatory funnel containing crushed ice and extracted with three 25 ml quantities of cold ethyl acetate. The cold Athylaeetatextrakte are combined, washed with cold water, cold 10biger sulfuric acid, cold aqueous sodium carbonate and cold V / washed, then evaporated ater over sodium sulfate and potassium carbonate and dried under reduced pressure at less than 4O ⁰ C to give a residue remains, the bis- the Mesylate of 6-endo- (1 ', 2' -dihydroxyhepty ^ - ^ - io ^ earbomethoxyhexylJ-bicyclo- ^ ji »0 / -hexan-3-one. This residue is dissolved in 4 nil acetone.

"Dissolves, then 2 ml of water are added, and the mixture is turned off for about 18 hours at room temperature, then it is diluted with 5 ml of water and extracted with three 30 ml quantities of ethyl acetate. The ethyl acetate extracts are combined with 5 ml of saturated washed aqueous Natriunbicarbonat, evaporated over sodium sulfate and dried under a reduced pressure at below 40 ⁰ C; as a residue is obtained 0.102 g, product containing PGE ^ raethylester This residue is chron over 15 g silica gel;. atographiert with 15 nl fractions Solvent eluted as follows:

Solvent fraction

1-6 £ 25 ethyl acetate - £ 75 Skellysolve B. 7-11 5Ό) ί. "£ 50" °

.12-16

17-21

22-24 '. - 95 & ■ "'■. 5% methanol

The eluted fractions were evaporated and analyzed by thin layer chromatography (ethyl acetate on silica gel) * Fractions 17-19 (total weight: 19 mg) consist of PGE ^ methyl ester. Fractions 14-16 (total weight: 23 mg) consist of 15-epi-PGEi-diethy-lester with a little moiioinesylate. Fractions 10 and 11 (total volume: 12 mg). "Consists mainly of PGAj-methyl-ester, and fraction 9" (13 mag): consists of .1.5 ^ 'epi-P.GÄ ^ -methylester .

0Ö9827 / 2053

Example 23 Endo-bicyclo / ^ j 1, oJ-hexan-S-ol-ö-carboxylic acid-. methyl ester *

A mixture of 103 g Endo-bicyclo - / 5.1. , oT carbonsauremethylester and 650 ml of anhydrous diethyl ether is stirred under nitrogen and at ⁰ -5 C gekühlt.-2-84 ml. a 1 molar diborane solution in tetrahydrofuran is added dropwise over a period of 30 minutes, the temperature being kept below ⁰ ° C. The mixture is allowed the resulting mixture under stirring to warm .while 3 hours at 25 ⁰ C. Evaporation under reduced pressure gives a residue which is dissolved in 650 ml of anhydrous diethyl ether. The solution is cooled to O ⁰ C, and 3 η aqueous Natriumhydroxydlösting (172 ml) is added dropwise ·-under nitrogen with vigorous stirring for 15 minutes while the temperature is maintained at 0 ° to 5 ⁰ C. Now 94 ml 3.0 # weight aqueous hydrogen peroxide are added dropwise with stirring during 30 minutes at 0 ° to 5 ⁰ C was added. The resulting mixture / v ird stirred one hour while it warmed to 25 ⁰ C check. 500 ml of saturated aqueous sodium chloride solution are added and the diethyl ether layer is separated off. The aqueous layer is washed with four 20G ml quantities of ethyl acetate, the washing solutions are added to the diethyl ether layer, which is then washed with saturated aqueous hatriochloride solution, dried and evaporated to give 115 g of residue. This residue is distilled under reduced pressure and gives 69 g of a mixture of methyl esters of endo-bicyclo- £ 5> 1, oJ-hexan-3-ol-6-carboxylic acid and endo-bicyclo- ^ 3,1,0 "j-hexan-2-ol -6-carboxylic acid; Boiling point: 86-95 ⁰ C at 0.5. mm. ·

- ■ Example 24 Endo-bieyclo - / 3.1 _r o7-hexane-3-ol · -6-carbQnsäure - - Etethyle st he ~ t et rahydropyrany lather

66 g of Geralschs obtained in Example 23, from 2-01 and 3-01 in 66 ml Dihydropropan be at 15 - 20 ⁰ C stirred and cooled while: 3 en si anhydrous _t with hydrochloric ge-r

009827 / 2D53

saturated diethyl ether can be added. The temperature of the mixture is kept ⁰ C for one hour under cooling in the range of 20 ° to 3O, then 15 hours long "allowed to stand at this temperature. Evaporation gave a residue which is distilled at reduced pressure is obtained. This gives 66 g of a mixture of the methyl ester. Tetrahydropropanyläther of Endo-bl.cyclö- / 3,1L, Oj-hexane-3-οΙ-β-carboxylic acid and Endo-bicyclo - / 3,1, Oj-hexane-S ^ ol- O-carboxylic acid with a boiling point of 96-104 ° C at 0.1 mm.

Example 25 Endo-6-hydroxymethyl-bicyclo- | J ~ 3, 1,0 / -hexan-3-ol-3-t "ethrahydropyranyl ether

A solution of 66 g of the mixture of the products obtained according to Example 24 in 300 ml of anhydrous diethyl ether is added dropwise over 45 minutes to a stirred and cooled mixture of lithium aluminum hydride (21 g) in 1300 ml of anhydrous diethyl ether under nitrogen. The resulting mixture is stirred for 2 hours at 25 ⁰ G and then cooled to 0 C. 71 ml of ethyl acetate are added and the mixture is stirred for 15 minutes. 235 ml of water are now added and the diethyl ether layer is separated off. The aqueous layer is washed twice with diethyl ether and twice with ethyl acetate. A solution of Rochelle salts is added to the aqueous layer, which is now saturated with sodium chloride and extracted twice with ethyl acetate. All diethyl ether and ethyl acetate readings are combined , washed with saturated aqueous sodium chloride solution, dried and evaporated; 61 g of a mixture of the 3-tetrahydropyranyl ethers of endo-6-hydroxymethyl-bicyclo-

3,1,0 / -hexan-3-ol and endo-ö-hydroxymethyl-bicyclo- / 3,1,0 / -hexan-2-ol.

Example_26 Endo-bicyclo- / 3,1,0? -Hexan-3-ol-6-carboxaldehyde-

3-tetrahydropyranyl ether

A solution of the mixture (34 g) obtained in Example 25

9S27 / 20S3 g _AD ORIGINAL

Products, in 1 000 ml. Acetone is cooled to -1O ° C. Then Jones reagent (75 hl a solution of 21 g of chromic anhydride, 60 ml of water and 17 ml of concentrated sulfuric acid), which to ⁰ ° C had been cooled in advance, was added dropwise with stirring for 10 minutes at -10 ⁰ C. After stirring for an additional 10 minutes, 35 ml of isopropyl alcohol was added over 5 minutes and stirring was continued for 10 minutes. The reaction mixture is then poured into 8 liters of an ice-cream mixture. The mixture obtained is extracted six times with dichloromethane. The combined extracts are washed with an aqueous sodium bicarbonate solution, dried and evaporated; 27 g of a mixture, the tetrahydropyranyl ether of Bndo-bicyclo-JJ, 1 »0 are obtained. -hexan-3-01-6-carboxaldehyde and endo-bicyclo- / 3,1,0 / -hexan-H-ol-e-carboxaldehyde hyd. - ";■; ■ -"■■■;"; ./■ - / \;:

l ^e j ^ Pi- ⁶ LL ^ 7 Endo-6- (cis- and trans-l-heptenyl) -bicyclo-3,1, ö / -hexan-3-ol-tetrahydropyranyl ether

A mixture of 100 g of hexyl bromide, 160 g of tripheylphosphine and 300 '-' ml of toluene is refluxed for 7 hours while stirring on the reflux condenser. The mixture is then cooled to 10 ⁰ C, and the separating crystals are filtered, washed with toluene and dried "wherein nan Hexyltriphenylphosjphoniumbronid 147 g having a melting point of 197 200 C ^0. \.

A mixture of Hexyltriphenylphosphoniumbrociid (102 g) and benzene (1200 ml) is stirred under nitrogen while a solution of butyllithium in hexane (146 ml of a 15 ^ strength Gew./) vol solution) was added v / ill. The resulting mixture is stirred for 30 minutes. A solution of 27 g of the mixture of the products obtained according to Example 26 in 300 ml of benzene is added dropwise with stirring over a period of 30 minutes. The mixture is heated and stirred for 2.5 hours at 70 ⁰ C, then is cooled to 25 ⁰ C. The precipitate obtained is filtered off and washed with benzene. The FiItrat and washing,

\: -: ■■ - · : - ^Λ 00Β827 / 2053

benzene are combined. - washed with water and dried, 53 g of one. Mixtures of the tetrahydropyranyl ethers of Endo-6- (cis- and trans-i »-heptenyl) -blcyclo-£ 5.1,0 / -hexan-3-ol and Endo-6- (cis- and trans-1-heptenyl) -bicyclo- £ 3,1,6] -hexan-2-ol receives. . >

Example 23 Endo-6- (cis- and trans-1-heptenyl) -blcyclo-

^: /.3,1,oJ -hexan-3-ol,

3 g of oxalic acid are added to a solution of 58 g of the mixture • of the products obtained according to Example 27 in 1500 ml of methanol. The mixture is refluxed with stirring for 1.5 hours. After evaporation under reduced pressure, an oil is obtained which is dissolved in dichloromethane. This solution is washed in an aqueous sodium bicarbonate solution, dried and reduced under reduced! Pressure evaporated. The residue is dissolved in a hexangene mixture (Skellysolve 15) and 600 g of moist-packed silica gel are chronographed. The column is filled with 2 1 Skellysolve B and then one after the other pit Skellysolve, first 1 1 rxj-t 2.5%, 2 1 with 5 # ", 2 1 with 7.5? Ί, 5 1 with 10 $ and 3 1 with 1 5% ethyl acetate in Skellysolve B eluted. After evaporation of the combined! traction, corresponding to the 10 and 15% ethyl acetate to 16 ^ of a mixture obtained 'from endo-6- (cis and Xvzcnz-I- heptenyl) -bicyclo- / 3,1, oJ-hexan-3-ol and Endo-6- (eis- 'and trans-l-heptenyl) -bicyclo- / 3,1, Oj-hexan-2-ol.

Example 29 Lndo-6- (cis- and trans-l-heptenyl) -bicyclo- /? »^ Ιo7 ~ hexan-3-one -

A solution of 15 g of the mixture of the products obtained according to Example 28 in 450 μl of acetone is ^{cooled to -10 0} C and stirred, while 30 ml of Jones reagent (Example 26) during. 10 minutes are added dropwise. The resulting mixture is stirred for 10 minutes at _T / -10 ⁰ C. Isopropyl alcohol (15 ml ·) is now added, stirring is continued for 10 minutes. “The mixture - .. is poured into 2400 ml of water. The water is used 5 times

0Q9827 / 2053

1137101

- so - λ; ■■ λ. ■ .. ■ ■ .... ν '

Dichloromethane extracted. The combined extracts are washed with an aqueous sodium bicarbonate solution _? - dried and evaporated, and an oil is obtained which is chromatographed on 50Q g of silica gel, which is packed moist with hexane ^ isomer mixture (okellysolve B) and successively with 2 1 Skellysolve B, 2 1 2, Seigern ethyl acetate in Skelly.solve B as well . 10 1 5 ^ -igen ethyl acetate in Skellysolve B is eluted. The first 1.5 l of the eluate with 5% ethyl acetate are evaporated; -und jr.an receives 5.9 g of Endo ^ 6 - (cis- and trans-1-heptenyi) -bicyclo - / ^,!, ol-hexan-3-one; Rf ~ 0.62 for the thin film -. Chromatography on silica gel plates ,, which with 20 ^ Äthyl- ™. acetate developed in Cyclohexaii '^ rden. .

Following the procedures of Examples 27, 28 and 29 except that ■ in example 2? Butyl bromide, pentyl bromide, heptyl bromide and Octyl biomide can be used instead of hexyl chloride one the. 1-pen% iyl-, 1-hexenyl-, 1-octenyl- and 1-NOnenyler-. "bonds corresponding to the product of Example 29"

: Following the procedure of Examples 27, 28 and ? 9, but where, in Example 27, primary bromides of the formula X- (CHp) ^ OIUBr are used - """v / ends v / crlen, ir; 5 series. i equals 1, 2, 3 or 4 and X isobutyl, 'tert-butyl, I' / ii-diiluortutyl, 4,4-difluorobutyl and 4 , 4,4-. '. ■ ■■■ -trifluorobut / .J. 'instead of-.von- hexyl tromide, one obtains compounds, c.iei ö.o ^r . Product of the 3eis "ielE 29 ontsprcchen, v.'C with X- (CH ^) --CH = CH- is in place of the 1 -Hepteny! Group.

ilacli the version of Examples 27 , 28 and 29, but in Boispiol 27, jev / e-ils: the iJxo-Blcyclo- £ 3,1, 07-lic.x-un Vor.,; bindings -, nc-tcl-lo Cor in example 27 specified Bndo- ■ connections .. used. -weredt-n, according to example 29 the "de :; end-product diei-es example en ^f t speaking en Ξζο- Compounds recovered. Jie n'jtv / endlgen Er.o-nicyclo- ß, 1, Oj -hexane compounds v / crder: according to; Belgian patent 702 744 er.: ■■ iialten, - ■■ '-. " ■■■. * ■ '■ ",": ■ ... . ^; .

BADOBIÖINAU 009827/2053 ,, ™ _ -

Example 30 Endo-6- (cis- and "" tr_anp-1-octenyl) -bicyclo -J3-, 1,0 / -hexan-3-ol-tetrahydropyranyl ether

A mixture of heptyl bromide (100 g)., TriphenylphcGphin (150 g) and toluene (300 ml ") are stirred for 7 hours heated on the reflux cooler. The mixture is now cooled to 10 C, and the crystals that separate are filtered off, washed with toluene and dried to give heptyltriphenylphosphonium bromide received lt.

A mixture of 105 g of heptyltriphenylphosphoniuin bromide and 1200 ml of benaol is stirred under nitrogen while a solution of butyllithium in hexane (146 ml of a 1-5% solution - w / t) is added. The solution is stirred for 30 minutes. A solution of 26 g of the mixture of the products obtained according to Example 26 in 100 ml of benzene is then added dropwise with stirring over 30 minutes. The mixture is heated for 2.5 hours at GO - stirred 70 ⁰ G and then cooled to about 25 ° C "The resulting precipitate is filtered off and washed with a little benzene filtrate and washing liquid v / combined ground, three times with 250 ml.. Washed quantities of water and dried over sodium sulphate. The Bep.sol solution obtained is evaporated to dryness, and 40 g of a mixture of tetrahydropyranyl ethers of endo-6- (cis- and trans-loctenyl) -bicyclo ~ f3,1 are obtained, Oj -hexan-3-ol and Endo-6- (cis- and tranti-l-octenylj.-bicyclo- / 3,1, oy-hexan-2-ol.

Example 31 Endo-6- (cis- and trans-l-octenylj-bicyclo- / 3,1, Oj -hexan-3-ol

ο 1.5 g of oxalic acid become a solution of 40 g of the mixture

the products obtained according to Example 30 in 700 ml of methanol given. The mixture is stirred for 1.5 hours on

-j reflux condenser heated. After evaporation under reduced

_iN) pressure gives an oil which is dissolved in 400 ml dichlorometha.n ·

° will. This solution is mixed with an aqueous sodium bicarbonate

CTt ' "- ■

Washed up sodium solution, dried over sodium sulfate and under evaporated under reduced pressure. The residue (31 g) is v / ird in

100 ml of a hexane mixture (Skelly & Olve ~ B) dissolved and chromatographed on 600 g of moist-packed silica gel. The column is eluted with 2 liters of Skellysolve B and then successively with 1 liter of Skellysolve B with 2.5%, 5 liters with 10% and 3 liters with 15% ethyl acetate. After evaporation of the combined fractions, with 10 % and 15% ethyl acetate, 15.5 g of a 'mixture of Endo-6- (cis- and trans-l-octenyl) -bicyclo - / _ 3,1, OJ-hexifji- 3-ol and Endo-6- (cis- and trans-octenyl) -bieyclo- Ϊ31 1, 0 ■; -hexan-2-ol.

Example 32 Endo-6- (cis- and trans-1-octenyl) -bicyclo-13,1, OJ-hexan-3-one .

A solution of- 15.5 g of the mixture of products obtained according to Example 31 in 450 ml of acetone is cooled to -10 ⁰ C and stirred while 30 Jones reagent (Example 26) was added drop- ml ■ as v / hile 10 minutes are kept, the temperature between -10 ° and O ⁰ G is kept. The stirring will

Addition of the Jones reagent is continued for a further 10 minutes, and then 15 ml of isopropyl alcohol are added and the mixture is stirred for a further 10 minutes. The mixture is poured into 2.5 l of water. The water is extracted with five 500 ml quantities of dichloromethane. The combined extracts are grounded with aqueous sodium bicarbonate, dried over sodium sulfate and evaporated to give an oil. This oil is dissolved in 1-00 ral isomeric hexanes (Skellysolve B) and chromatographed on 500 g of silica gel which is packed moist with Skellysolve B. The column is filled with 21 Skellysolve 3 and then successively with ethyl acetate _; Holding Skellysolve B 2 1 aj-t 2.5% and 8 1 with 5% Athylace- , tat eluted. The first two liters of eluate with 5 ^ ethyl acetate are evaporated, and 4.8 gl? Ndo-6- (cis- and .trans-1 --- octenyl) -bicyclo- "£ 3, 1, Oj -hexane are obtained -3-on.

Example 33 Methyl-6-endo- (1-octenyl) -3-oxo-bicyclo- / 3,1, P / -hexane-2-heptanoate

A solution of 4.8 g of Endo-6- (ciQ and. Trans-l-octenyl) - bicyulo- ^

BATHROOM ORIGINAL - r

3,1,0 -hexan-3-one from Example 32 and 12.7 g of 7-iodo-heptanoic acid methyl ester in 75 ml of tetrahydrofuran are prepared, and nitrogen is bubbled through the solution for 5-10 minutes. A solution of 3.91 g of potassium tert-butoxide in. 150 ml of tetrahydrofuran is also flushed through with nitrogen. The two solutions are then simultaneously added dropwise at 25 ° C. to one end of a 70-80 cu long horizontal raw material for 45 minutes admitted. The reaction mixture drips from the tube in a 40 nl 5 / '> hydrochloric acid-containing flask, the mixture is b'ei under reduced pressure in a bath 40 - 50 ⁰ C concentrated, ur .; the greetings. Remove part of the tetrahydrofuran. The residue is diluted with 100 ml of water and then extracted with four 100 ml of ethyl acetate. The first three amounts of ethytes are combined and washed with 5% aqueous sodium thiosulphate solution and then with aqueous saturated sodium chloride solution. The wash liquors are extracted back with the fourth ethyl acetate extract. The ethyl acetate extracts are now combined, dried over anhydrous sodium sulfate and evaporated under reduced pressure, giving an oil. The whole raw Cl is dissolved in Skellysolve B and chronographed over JCG g of clay (type II). The column is nit steisende amounts Eenncl Skellysolve B containing 1.5 * 1, C eluiort with 10fi, 1.5 1 nit 20 f> 1.4 and 1 nit 50 $ benzene and schlieiS-lich nit i 1 benzene. The £ 10 and £ 20 Benzel S: ceiry-solve B eluates are evaporated, and 12.55 g of a mixture of 7-I.pd-heptanoic acid ethyl ester and Aus ~ ->. Ngsketone are obtained The last 1000 g of the 50% benzene eluate and the benzene eluate are also evaporated, and 1.192 g of Cl are obtained. This Cl is dissolved in Skellysolve B and chronographed over 150 g of silica gel. The column is filled with 750 ml of Skellysolve B. and eluted in succession with 750 nl 2.5% -> 30000 ml of 5% iind 750 nl 10% ethyl acetate in Skellysclve B •, a first fraction of 750 nl Skellysclve B and then 150 nl fractions being removed. Actions 11 through 15 are evaporated and pooled , and msii is obtained

0098? 7/2053

- 84 - '■■;■' ■. ■ '- ■;.: ·.

0.62 g of methyl 6-endo-6i-octenyl) -3-oxo-bicyGlo-£ 3.1 »ol-liexane-2-heptanoate (weaker polar isomer). Fractions 16 "bis are combined and give 0.238 g of methyl 6-endo (1-octenyl) - ^ -oxo-bicyclo- ^, 1, oJ-hexane-2-heptanoate (more polar isomer). -. ■ _r "-".. ■

Example 34 Methyl 6-endo- (1-octenyl) -3-oxo-bicyclo- ^ 3,1,07-hexane-2-heptanoate

A solution of 3.05 g of potassium t-butoxide in 400 ml of tetrahydrofuran v / i rd dropwise with stirring under nitrogen at 25 ° C. over a period of 45 minutes to a solution of 3.75 g of Endo-6- (cis- and traris-l-octenyl:) - bicyclo ~ / 3, ϊ, θ] - ■ '■'. ' hexan-3-one and 14.7 g of 7-lQd-heptanoic acid diethyl ester in 200 ml of tetrahydrofuran. The reaction mixture is stirred about 15 minutes after the addition of the butoxide solution has ended and 40 ml of 5% hydrochloric acid are now added. This mixture is diluted with 150 ml of water and extracted with four 100 ml quantities of ethyl acetate. The first three ethyl acetate extracts are combined, washed with 5% aqueous sodium chloride sulfate and then with saturated aqueous sodium chloride solution. The fourth ethyl acetate extract is as Rückwaseh- ■ _t -, medium used .. The ethyl acetate extracts are combined, evaporated over Fatriumoulfat dried under reduced pressure to "wol egg will receive oil This crude 01 v / ill in 50 ml Skellysolve B solved?. and cliromatographed over 300 g of clay (type II). The column is filled with 1.5 1 of Skellysolve 3 and then successively with 1.5 1 of 205% and 1.5 1 of 5 % Bensol'in Skellysolve B and finally with 1, Washed 5 liters of benzene. The 50 ^ ige benzene in "Skellysolve B and the first 300 ml of the benzene eluate are evaporated, and 413 g of 01 are obtained. This oil is dissolved in Skellysolve 3 and. chromatographed on silica gel. The column is eluted with 750 ml Skellysolve B, then with 750 ml 2, Seigern and 3000 ml 55iigen Ätliyla.eetat in Skellysolve B, with a fraction of 750 ml, 450 ml and then one after the other from 150 ml.

OK

will catch. Fractions 9-12 are evaporated and ~ agrees, and one receives 0.866 g of methyl-6-endo- (1-octenyl) -3-oxo-bicyclo-r "3, 1, Of-hexane-2-heptanoate (weaker polar Isomer). Fractions 13 "to 20 are injected and combined, 'and 0.312 g of methyl-6-endo- (1-octenyl) -3-oxo-bicyclo- / * 3.1 are obtained , 0] -liexane-2-heptanoate (more polar isomer).

Example 35 methyl-6-endo- (1,2-dihydro'xyoctyl) -3-oxo-bi-

cyclo- £ 3,1,07-hexane-2-heptanoate

A solution of 1.5 g of methyl-6-endo- (1-octenyl) -3-oxo-bicyclo-T "3.1 , Ö] -hexane-2-heptanoate (weaker polar isomer from example 33 and Example 34) and 1.3 g of osmium tetraoxide in 30 ml Pyridine is stirred for 15 hours at 25 ° C; Now a solution of 3.6 g of sodium bisulfite in a mixture of 60 ml Water and 39 ml pyridine are added and the stirring is stopped for about Continued for 5.5 hours. This mixture is made with 100 ml v / ater diluted and extracted with three 400 ml quantities of chloroform. The chloroform extracts are combined with 100 ml of water washed, dried over sodium sulfate and evaporated under reduced pressure; 1.56 g of an oil are obtained. This oil is dissolved in 40 ml of 40% ethyl acetate in Skellysolve B. and over 150 g of silica gel chromatograph. The column is with 2.1 1 40 $ and 1.5 1 50% ethyl acetate in Skellysolve B eluted with 150 ml eluate fractions collected will. The weaker polar erythro-glyeol contained in fractions 6-8 is obtained in an amount of 0.644 g. The amount of the more polar glycol obtained in fractions 9-16 is 0.712 g. . .

Example 36 Methyl 6-endo- (1,2-dihydroxyoctyl) -3-oxo-bicyclo- ^ 3,1,0J-hexane-2ß-heptanoate

A solution of 0.55 g of methyl-6-eiido- (1-octenyl) -3-oxo-bicyclo ~ / j3.1 > 0 / -hexane-2-heptanoate (more polar isomer from Examples 33 and 34) and 0.43 g of osmium tetroxide in 10 ml.

00982 7/20 53

Pyridine is stirred for about 15 hours at 25 ^° C .; a solution of 1.2 g of sodium bisulfite in a mixture of 20 ml of water and 13 ml of pyridine is now added, and the process is continued for 5-6 hours. This mixture is diluted with 40 ml of water and extracted with three 140 ml quantities of chloroform. The chloroform extracts are combined, washed with 40 ml of water, dried over sodium sulfate and evaporated under reduced pressure, whereby 0.54 g of methyl-6-endo- (1,2-dihydroxyoctyl) -3-oxo-bicyclo- £ 3.1, Oj-hexane-2-heptanoate are obtained.

For example 37 _{20-methylprostaglandin-E]} methyl ester and 15-epi-20-methylprostaglandin-E] -methyl ester

A solution of 0.63 g of methyl 6-end0- (i, 2-dihydroxyüctyl) -3-oxo-bicyclo- / 3,1, OJ-hexane-2rK.-heptanoate (weaker polar glycol, fractions 6-8 of Example 35) in 20 ml of pyridine is stirred under nitrogen and cooled ρκβρτι witrerrdd in an ice bath. 2 ml methanesulfonyl chloride are added, and the solution is stirred in the melting ice bath for 2.5 hours. The solution is diluted with 30 ml of ice and water, stirred for TO minutes and then poured into a crushed 21s containing separating funnel given. The mixture is with extracted three 100 ml quantities of cold ethyl acetate. The ethyl acetate extracts are combined and with 70 ml of cold, 10 / ί-iger Sulfuric acid, then with cold aqueous sodium bicarbonate solution and water and dried over sodium sulfate and potassium carbonate for one hour, then evaporated. 0.89 g of dismesylate is obtained as an oil, which in 36 ml Tetrahydrofuran is dissolved, diluted with 12 ml of water and placed at room temperature for about 20 hours. The mixture is diluted with 25 ml of water and under reduced pressure concentrated to remove the tetrahydrofuran. The mixture is then diluted with 50 ml of water and with three 100 ml quantities Ethyl acetate extracted. The ethyl acetate extracts are combined and with aqueous saturated sodium bicarbonate solution

QQ9827 / 2Q53

and washed twice with saturated aqueous sodium chloride solution, then dried over sodium sulfate and evaporated to dryness to give 0.63 g of oil. This oil is dissolved in a mixture of 25% ethyl acetate in Skellysolve B and chromatographed over 50 g of silica gel. .The column is filled with 400 ml of 2 tigers. 250 ml of 50% and 250 ml of 75% ethyl acetate in Skellysolve B, then with 250 ml of ethyl acetate and finally with 250 ml of ethyl acetate containing 5% methanol, eluting first two 150 ml fractions and then 50 ml -Practions are accepted. Fractions 20 and .21 (- *; rhy lace tot, which contains 5% methanol) are evaporated, and 69 mg of 20-methylprostaglandin-E .. methyl ester are obtained. Fractions 14 to 16 (75 ^ ethyl acetate in Skellysolve B then two iitiiylacetatfraktionen). are evaporated to give 9.7-Jng; T5-epi-20-methylprostaglandin-E ₁ -methyl ester.

The more polar glycol (0.70 g, fractions 9 to 16 in Example 35) is treated with methanesulfonyl chloride, then solubilized and worked up as described above; one obtains C, € 9 ^ oil which, as described above, is chromatographed to give 139 mg of 20-methylprostaglandin-E. methyl ester and 125 ag i5-epi-20-methylprostaf: landija-Eu-methyl ester, see below.

"The off" obtained the 'Gixromatogrammen of the two experiments described above 2NC-Meth3 lprostaglandin ^r-S. -laethyl esters are vexeinrigt and ^ v ^ once crystallized from a mixture of ether and Skellysolve B, whereby an analysis-pure sample of 20-methylprostaglanäin-E.-methyl ester is obtained; Selinielz point ": 67-63 ° C; mass spectra: peaks at 382, 364, -546, 333,> 15, 314, 297, 293, 279, 247 and 204.

_{The i5-epi-20-ffiethylprostaglsndin-S 1} -inethyl esters obtained from the chromatograms of the two yersucles described above are combined and crystallized from a mixture of ether and Skeilysole B ^^ to give 1 5-epi-20-methylpröstagiandin-B. -methyl ester to give '. "./.-.-."

009827/1053

Example 38 \ ": B-iso-2O-methylprO'Staglana ^- -methylei3t in E ₁ ': ER _{r i} __und --S-iso-1 5-epi-20-methylprostaglandin E ^ ~ methylester

According to the procedure of Example 37, 0.54 g of methyl 6-endo- (1,2-dihydroxyoctyl) -3-oxo-bicyclo- ^ 3 ·, 1, oj-hexane-2ßheptanoat (obtained according to Example 36) with methanesulfonylchlo , -rid treated in "pyridine" and worked up '' is obtained 0.46 g of dimesylate, which was dissolved in 20 ml of acetone, diluted with 12 ml of water and is switched off for about 20 hours at 25 "C ^0. the mixture is diluted with 25 ml Water is diluted and concentrated under reduced pressure to remove the acetone and then extracted with ethyl acetate; the extract is washed, dried and concentrated as described in Example 37 to give 0.31 g of oil dissolved in 20 ml of 2-sigment ethyl acetate in Skellysolve B and chromatographed over 50 g of silica gel eluted with 2-50 ml of ethyl acetate and 250 ml of 5% methanol in ethyl acetate. An eluate fraction of 200 ml is first removed, then five 100 ml fractions and finally 50 ml fractions. . ";. '' .. '■ -

Fractions 14 to 16. (Ethyl acetate, then 5 ^ -iges methanol in Ethyl acetate) are evaporated "and ^^ combined, ^ and one obtains 29 mg S-iao-SO-methylprostaglaridin-E ^ -methylester, fractions ■ ". ' 8 to 12 (75% ethyl acetate in Skellysolve B, then ethyl acetate) v / are also evaporated and combined, and 51 mg of 8-iso-i 5-epi-2O-Meth3rlprGStaglandin-E.-methyl ester are obtained.

Example · 39 · Meth: yl-: 6.-endo- (7-meΐhyl- ■ l.-Ό.cteπyl) ■ -3-oxo-bicy ■ cl ■ o- ". ^ 3,1, Oj-hexane 2-heptanoate

Following the procedure of Examples 30, 31, 32 and 34, but using l-bromo-6-methylheptane instead of 1-bromoheptane in Example 30, i-ethyl-6-endo- (7th) is obtained from the last chromatogram -me thyl-l ^J ~ octenyi) -3-oxo-bicyclo- 3.1, 0 -hexan-2-heptanoate in the form zv / eggs isomers: a weaker

... Q 0. ^ 827/2053 bad

j 1337908

and a more polar isomer.

Example 40 ■ Methyl-6-endo- (7-methyl-1,2-dihydroxyoc-tyl) -3-

oxo-bicyelo- jj> , 1, OJ "hexane-2-heptanoate

A solution of '1.0 g of methyl 6-endo- (7-methyl-1-octenyl) -3-oxo-bicyclo- / 3,1, Oj -hexane-2-heptanoate (weaker polar isomer, which according to Example was obtained 39) in 13.5 ml of tetrahydrofuran is heated to 5O ⁰ C and a warm solution of 530 mg potassium and 35 mg of osmium tetroxide in 6.5 ml of V / ater is added with stirring, the mixture 5 hours at 50 ⁰ C., then it is concentrated under reduced pressure to remove the tetrahydrofuran. The mixture is diluted with water and extracted three times with dichi arme than. The dichloromethane extracts are combined, washed with water, dried over sodium sulfate and evaporated under reduced pressure. 1.0 g of oil is obtained, which is chromatographed over 120 g of silica gel. The column is filled with 500 ml of 10%, 1000 ml of 25%, 1000 ml; 35 ^ -igeDi, 1000 ml 45 ^ -igem, 1000 ml 5Obigem and 1000 ml 60 ^ -igem ethyl acetate eluted in Skellysolve B. The 35 ^ -ige ethyl acetate is concentrated, and 255 mg of the weaker polar form of methyl-6-endo- (7rraethyl-1,2-dihydroxyoctyl) "3-oxo-bicyclo- ^ 3,1, Oj-hexan 2-heptanoate. The 50% ethyl acetate is concentrated, and 248 mg of the more polar form are obtained. · '

Example 4 1 20,20-Dimethylprostaglandin-E ^ -methylester and

1 5-epi-2O, 20-dimethylprostaglandin-E. -methyles ^ -er

A solution of 0.255 g of methyl ~ 6-endo- (7-methyl-1,2-dihydroxy " octyl) -3-oxo-bicyclo-3,1,0 -hexane-2-heptanoate (weaker polar glycol, which was obtained according to Example 40) in 7 nil Pyridine is stirred under nitrogen while cooling in an ice bath and 0.7 ml of methanesulfonyl chloride are added admitted. Stirring is continued for 2.5 hours. The ■

00.982 77 20 S3- '

.- 90 -

The solution is diluted with 30 ml of ice and water and stirred for 10 minutes, then it is poured into a separating funnel containing crushed ice and extracted three times with 100 ml of ethyl acetate. The ethyl acetate extracts are combined, washed with cold 10% sulfuric acid, cold 10% sodium carbonate and ice water, then dried over sodium sulfate and evaporated. 338 mg of Dime sy lat are obtained as an oil, which is dissolved in 8 ml of acetone, diluted with 4 ml of water and ^{stored at 25 °} C. for about 20 hours. The reaction mixture is now diluted with 25 ml of water and concentrated under reduced pressure; then 50 ml of water W are added and the mixture is extracted three times with ethyl acetate. The ethyl acetate extracts are combined, washed with saturated aqueous sodium carbonate solution and saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated; 258 mg of an oil are obtained.

After the above procedure, but using the more polar glycol (248 mg, prepared according to Example 40) as the starting material, 270 mg of an oil is obtained which, as was determined by thin-layer chromatographic analysis, is identical to the oil which was obtained from the weaker polar G-Iycol. These two oils are combined (528 mg) and over 70 g of silica gel -chromate -'-. The column is filled with 0.6 1 205%, 1 1 35%, 1 1 40%, 1 1 50% and 3 1 75% ethyl acetate in ' okellysolve B, then eluted with 1 l of ethyl acetate and 11 5/1 strength MeOH in ethyl acetate, with 75 sil fractions being removed. The eluate fractions 67 to 73 are evaporated and combined, and 64 mg of 15-epi-20,20-dimethylproate / aglandin E. methyl ester are obtained; IR absorption at 3440, 1740, 1250, 1200, 1165, 107 and 970 cm " ¹ .

The eluate fractions 88 to 104 are evaporated and combined, and 111 mg of 20,20-dimethylprostaglandin-E ₁ methyl ether are obtained. This is made of a mixture of ether and

Skellysolve B is crystallized to give an analytically pure sample of 20,20-Dimethylprostaglandin-E .. having a melting point at 75 - 76 ⁰ C is obtained; Mass spectrum: peaks at 378, 360, -347, 297, 279 and 218; IR absorption at 3310, '1735, 1325, 1310, 1290, 1275, 1260, 1225, 1195, 1150, 1105, 1065 and 975 cm " ¹ .

Example 42 8-Iso-20,20-dimethylprostaglandin-E. methyl ester and S-Iso-15-epi-20,20-dimethylprostaglandin-E ₁ - 'methyl ester

Following the procedure of Examples 40 and 41, except that in Example 40 the more polar methyl-6-endo- (7-methyl-1-octenyl) -3-oxo-bieyclo-3,1,0 -hexane-2- heptanoate is used instead of the weaker polar isomers, 8-iso-20,20-diniethylprostaglandin-Ej-methyl ester is obtained; Mass spectrum: peaks at 396, "378, 360, 347, 297, 279 and 218. Rf; 0.47 after thin layer chromatography on silica gel with the A-IX solvent system, and furthermore 8-Tso-i5-epi-2G, 2C - dimethylprostaglandi "n ^ E ₁ -methylester; Mass spectrum: peaks at 396, 378, 360, 347, 297, 279 and 218; Rf: 0.36 on silica plate with the A-IX solvent system.

Example 43 19-Methylprostaglandin E .. methyl ester and

1 5-epi-1 g-methylprostaglandin-E.] Methyl ester

Following the procedures of Examples 30, 31, 32 and 34, but with in Example 30, 5-methylhexyl bromide instead of Heptyl bromide is used, is obtained from the last chromatogram Methyl-ö-endo-C ^ -methyl-1-heptenylJ-S-oxo-bic.Yclof5,1,6? -Hexane-2-heptanoate in the form of two isomers: a:

'L-j _

weaker and one more polar isomer. -

Following the procedure of Examples 40 and 41, except that in Example 40 the weaker polar isomer of methyl-6-endc- (6-ynethyl-1-heptenyl j -3-Qxo-bicyclo-73.1, θ] -hexane -2-heptanoate

009827 / 20S3 ^

instead of methyl-ö-endo - ^ - ynethyl-l-octenylJ ^ -oxo-bicyelo- "Γ3,1, Oj -hexane-2-heptanoate ^ is used, one receives 19-methylprostaglandin-E, .- methyl ester; . melting point: 52 - 53 ⁰ Gj Absörptioia IR (mull) at 3430, 3290, 1740, 1675 (weak), 1300, 1275, 1225, "-1200, 1170, 165 and 990 cm"^1; and 15-epi-1 g-methylprostaglandin-E ,. methyl ester; IR absorption at 3420, 1740, 1250, 1200, 1165, 1075 and 1035 cnT ¹ , 'mass spectrum; Peaks. At 382, 364, 351, 346, 297, 293, 279 and 247.

Example 44 19-methylprostaglandin A ^ -methyl ester and

A solution of 200 mg of 19-methylprOstaglandin ~ E. | methyl ester ■ -.in a mixture of 2 ml of tetrahydrofuran and 2 ml of 0.5 η hydrochloric acid is stirred under nitrogen at 25 ° C for five days. The reaction mixture is thinned with saturated aqueous sodium chloride solution diluted and extracted with ethyl acetate. The EthyIacetatextrakt is with saturated aqueous sodium chloride -gewäsclien, dried over llrLtriuinsulfat and evaporated. 159 g of an oil are obtained, which over 25 g of silica Chromatographed and mixed with 350 ml of 20%, 400 ml of 30%, • 5:00 ml 40 ^ igein, 1000 ml 50 ^ igexa and 500 ml 60 ^ igexa ethyl acetate in Skellysolve B, then eluted with 500 ml ethyl acetate taking 25 ml fractions. The eluate fractions 17-22 are concentrated and united, and - = one receives 45 mg ^ -Methylprostaglandin-A.-methyl ester. '

(Ethanol solution) -maximum at 217 ku *, Schvilter at 204. After heating with sodium hydroxide in ethanol, the UY maximum was at 278 mu, the shoulder at 235 n ^ X.iig-methylprotaglandin-B.-r-methyl ester). The eluate fractions 28-35 are concentrated and combined, and 25 mg of 19-methylprostaglendin-A ^ are obtained; IR absorption at 332Ό,. 172O and 1585 cm " ¹

009827/2053 bad original

Example 45

bicyclo-f3, 1, θ! -hexane-2-heptanoate

Following the procedure of Examples 30, 31, 32 and 34 "but in Example 30 l-bromo-6,6-dimethylhepta.n is used instead of - '"". ■ 1-bromoheptane is obtained from the last ohmic , matogram methyl 6-endo- (6,6-dimethyl-l-heptenyl) ~ 3-oxo-bi- • cyclo- / 3,1, OJ-hexane-2-heptanoate in the form of two isomers: one weaker and one more polar isomer.

Example 46 Methyl-6-endo- (6,6-dimethyl-1,2-dihydroxyheptyl) -3-oxo-bicyclo- [3,1-, ÖJ -hexane-2-heptanoate

A solution of 12.0 g of methyl 6-ehdo- (6,6-dimethy1-1-heptenyl) -3-oxo-bicyclo- [j5,1, θ] -hexane-2-heptanoate (weaker polar isomer, the was obtained according to example 45) in 150 ml of tetrahydrofuran is heated stirred .on 50 ⁰ C under nitrogen; then 1 g of solid osinium tetroxide and then immediately a warm solution of 6.5 g of potassium chlorate in 76 ml of water are added all at once. The reaction mixture is stirred for 5 hours at 50 ° C. under nitrogen; then it is concentrated under reduced pressure to remove the tetrahydrofuran. The mixture is diluted with water and three times with Diehlor-. methane extracted. The dichloromethane extracts are combined, washed with water, dried over sodium sulfate and evaporated under reduced pressure. 14.0 g of an oil are obtained which is chromatographed over 2 kg of silica gel. The column is eluted with 8 1 15%, 12 1 25 ^, 16 1 35 ^, 16 1 45 ^ 0 and 8 1 60% ethyl acetate in Skellysolve B, with 600 ml practical ions are removed. Frcictions 22 to 66 are evaporated and combined, and 9.0 g of methyl-6-endo- (6,6-dimethyl-T, 2-dihydrohept.yl.) -3-oxobicyclo-3,1,0 are obtained "hexane-2-heptanoate.

Example 47 19, ^ -Biriiethylprostaglandin-1Z-methyleBter and - 1 5-epi-19,19-dimethylprostaglandin-E ^ -methyl ester

A solution of 9.0 g of methyl-6-endo- (6,6-dimethyl-1,2-dihydroxy-

'009827/2053

1937808

heptyl) - ^ - oxo-bicyclo- J3,1, ol-h.exane-2-heptanoate (prepared according to'Be is.piel 46) in 110 ml of pyridine is stirred under nitrogen and cooled in an ice bath, while 10.7 ml of methanesulf onylehlorld are added dropwise over 15 minutes. The mixture is stirred 2.3 hours at 0 ° C, then a dry ice-acetone Baäes to -10 ° C to -1-5 ⁰ "/ cooled by means, whereupon 10 ml of ice-V / ater slowly added with vigorous stirring while the temperature is kept below ⁰ G .. The mixture is poured into 500 ml of ice and water

poured »Then 200 ml of a cold dichloromefchan-ether-mixture are poured (1; 3) and 440 ml of cold 3 η hydrochloric acid were added, and P. ", the mixture is quickly separated. The mixture becomes three

extracted further times with 200 ml quantities of a cold dichloromethane-ether mixture: (1: 3). The dichloromethane ethereal extracts are combined, washed with cold 2% sulfuric acid, cold 10% aqueous sodium carbonate solution and cold saturated aqueous sodium chloride solution, dried over sodium sulfate and potassium carbonate and evaporated. 14> 0 g of oil are obtained, which is dissolved in 450 ml of an acetone-water mixture (2: 1) and placed at 25 ^° C. for about 20 hours. The reaction mixture is diluted with 200 ml v / water and concentrated under reduced pressure to remove the acetone 2SU. Subsequently, v / ground 100 ml of water supplied _k, and the mixture is washed four times with Jthylacetat Extra-P. here. The ethyl acetate extracts are mixed with ^ aqueous! Sodium-. washed carbonate and aqueous sodium chloride over sodium sulfate; dried and evaporated. 9.5 g of oil are obtained, which is chromatographed over 1.6 kg of silica gel. The column is filled with 4 1 20 ^, 8 1 .3Obig, 8 1 40%, 20 1 βθ ^ and 20 1 80 ^ -igem ethyl acetate in Skellysolve B, y then with 20 1 ethyl acetate and 4 1 Eluted 5 ^ -igem methanol in ethyl acetate, 600 ml portions are removed. The inhaled fractions 66 to 72 are evaporated and combined, and 1.253 g of IS is obtained

IaIIdIn-B ₁ methyl ester; IR absorption at 3420, 1740, 1245, 1200, 1165, 1075, 1020 and 970 cm " ¹ .

0O9827 / 2OS3

The eluate fractions 96-111 are evaporated and combined, and 1.228 g of 19,19-diiaethylprostagla.ndin-E.-methyl ester are obtained. This is crystallized from a mixture of ether and Skellysolve B, and gives a product of melting point: 53-55 ⁰ C; IR absorption (cheesecloth) at 3450, 3390, 3280, 1740, 1675 (weak), 1310, 1290, 1275, 1235, 1195, 1165, 1105, 1090, 1065, 1020 and 985 cn "¹; mass spectrum: peaks at 390 , 386, 378, 372, 358 and 343.

Example 48 19,19-Dimethylprostagla & din-F .. and

19,1 9-Dimethylprostaglandin P ^ -.

A solution _: of 500 mg of 19,19-dimethylprostaglandin-E ^ -methylester in -25 ml of isopropanol is ^{stirred at 0} ° G under nitrogen, and a cold solution of 250 mg of lirtrium borohydride in 5 ml of water is added. The mixture is stirred at ⁰ C for 2.5 hours, then nl is 1 acetone was added and the mixture is stirred for a further ten Hinuten long ⁰ G at O. The mixture is made slightly acidic (pH 5-6) with acetic acid and then concentrated under reduced pressure to remove the acetone and isopropanol. The residue is poured into saturated aqueous sodium chloride solution and extracted three times with ethyl acetate. The A thy lac et at extracts are combined, washed with saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated. This gives 507 mg of a mixture of 19,19-Dimethy! Prostaglandin F ^ methylester and 19,19-Dimethylprostaglandin _F-1ß methyl ester as a white solid. This mixture (503 ng) is dissolved in 15 ml of methanol, cooled to about 5 ⁰ C and stirred under nitrogen while 2 nl 50% aqueous KaJLiunhydroxyd be added. The mixture is then stirred under nitrogen for four hours at 25 ° C., then diluted with 100 ml of water and extracted once with ethyl acetate. The aqueous phase is acidified with dilute hydrochloric acid and extracted four times with ethyl acetate. The ethyl acetate extracts are combined, three times with ' Water and once with saturated aqueous ■ '"■

009827/2053

■ .... ■■■: _ 96 ■. .. ■ ': ■■. / ■: '· ■ ./; ■'

gen sodium chloride -washed, dried over sodium sulfate and evaporated. -506 mg of a white crystalline are obtained Material that is chromatographed over 150 g of silica gel will. The column is filled with 500 ml 50 ^ and 500 ml 75 $ - igen ethyl acetate in cyclohexane then with 4000 ml of ethyl acetate and then with 500 ml of 10% and 500 ml of 25% Methanol eluted in ethyl acetate. The ethyl acetate-cyclohexane eluates are discarded, then 50 ml fractions are removed, starting with the ethyl acetate eluate. Political groups. 16 - 35 are evaporated and combined, and one obtains 135-rnc 1 9.1 9 * -Dimethyl prostaglandin- ^ ^, which consists of a Mixture of ethyl acetate and Skellysolve B recrystallized will. Men receives 19,19-dimethylprostaglandin-F ^ - from the melting point: 10? - 1 over 9 ° C; IH absorption at 3320, 2700, 1710, 1325, 1305, 1290 ,. 1275, .1240, 1210, 1200, 109.5, .1,050, 1020, 985, 975 and 945 cm "; mass spectrum spectral pits at 384, 366, 348 and 294.

Fractions 46 to 34 are evaporated and combined, and 211 mg of 19,19-dimethyl-PGF are obtained. _β , which is recrystallized from a mixture of ethyl acetate and Skellysolve B; 19.1 9-BiKiethylijrostag ^J landin-F _1β from 3 c lime la punk t % ^{145-146 0} C are obtained; IR absorption at 3360, 2700 ,. 1710, 1305, • 1290, '12?: 0, .1030, 1015, 995, 970 and 950 cm " ¹ .

Example 49 Methyl - 6-endo - (- 1 -heptenyl ^ - ^ - oxo-bicyclo-M, 1, o -hexane-2- (2, 2-diinethylheptanoate)

Its solution top 6.33 S 6-Endo- ('1-heptenyl) -bicyclo- / 3, 1, 0 ^ J-3-an (prepared according to Example 29) and 14.6 g MoUiyl-7- ^: iodine -2,2-cJiaethylbeptaiioat in 2OG ml of tetrahydrofuran is ^{stirred under nitrogen at 25 0} G, and a solution of 3.8 g of potassium t-butoxide in BOO nl of totrahydrofuran is slowly drawn for 45 minutes. obc-n. 70 ml of S # hydrochloric acid and then 5 nl of pyridine are added. The mixture is concentrated under reduced pressure to remove most of the tetrahydrofuran and diluted with 200 ml of ice water,

BAD ORKälNÄL 009827/2053 ~~ * '"'

The mixture is extracted with two 200 ml quantities of an ether-dichloromethane mixture (3: 1). The ether-dichloromethane solution is washed successively with dilute hydrochloric acid, water, dilute aqueous sodium thiosulfate and saturated aqueous sodium chloride. The washed solution is dried over sodium sulfate and evaporated under reduced pressure to give 16.9 g of oil which is chromatographed over 1.5 kg of silica gel which would be wet packed with 2% methanol in dichloromethane. The ■ column is filled with 6 1 dichloromethane, 6 1 1 ^ -igem. and 6 1 2tfo strength methanol in dichloromethane eluted, whereby. JOO ml practicals to be removed. Fractions 25 to 36 are evaporated and combined, and 4.25 g of methyl-6-endo- (1-heptenyl) -3 ~ oxo-bicyclo - ^ 3,1, o7-hexane-2 .-- ( 2,2-diniethylheptanoate) (weaker polar isomer).

Example 50 "Methyl-6 ~ endo ~ (i., 2-dihydroxyheptyl) -5-oxo-

bicyclo- ^ 3,1, o7-hexane-2- (2,2-dimethylheptanoate)

A solution of 10.38 g of methyl 6-endO ~ (1 ~ heptenyl) -3-cixobicyclbj_3,1, o7-hexane-2- (2,2-dimethylheptanoate) (weaker polar isomer, which was obtained according to Example 49) in 250 ml of tetrahydrofuran is ^{heated to 50 °} C. and stirred. Osmium tetroxide (0.5 g) is added, then a warm solution of 8.5 g \. Potassium chlorate in 100 ml of water, and the mixture is stirred for SWEi hours and 40 minutes at 50 ⁰ C. The mixture is concentrated by reduced pressure distillation to remove most of the tetrahydrofuran. The aqueous residue is extracted with dichloromethane. The dichloromethane extract is washed with water and an aqueous saturated sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure to give 14.1 g of an oil. The oil is chromatographed on 1,400 g of silica, which is packed wet with an ethyl acetate-cyclohexane mixture (1: 1). The column is · eluted with an ethyl acetate-cyclohexane mixture (1: 1), where '200 ml -]? Rak- ·

009827/2053

tioneh are collected, fractions 20 to 45 are evaporated and combined, and 7.3 g of methylene 1-6 end o- (1,2-dihydroxyheptyl) -S-oxo-bicyclo- f3 , Λ ,, θ7 -hexane-2- (2,2-dimethylheptanoate). Fractions 10 to 19 are also evaporated and combined and dissolved in 200 ml of tert-butanol. Sine solution of 2.5 g Natriuinhydrosulfit in 60 ml of water and 30 g "Magnesol" (Magensiumsilikat) are added and the Genisch is stirred for 30 minutes at 25 ⁰ C. The mixture is filtered and the filtrate is concentrated under reduced pressure to remove the tert-butanol. The residue from oil and water is extracted with dichloromethane, the extract is washed with aqueous sodium chloride, dried over sodium sulfate and evaporated under reduced pressure, with 2.36 g of oil being obtained. The oil is chromatographed over 200 g of silica gel. The column is eluted with a mixture of cyclOhexane and ethyl acetate (1>. 1, ml fractions being removed. Fractions 16 to 35 are evaporated and chromatographed, and a further 0.770 g of methyl-6-endo- (1, 2- dihydroxyheptyl) -3-oxo-bicyclo- £ 3.1, oJ-hexane-2- (2,2-dimethylheptanoate).

The total yield is 8.07 g. .

See Example 51 2,2-dimethylprostaglandin E.i methyl ester and

epi-2, ^ -dimethylprostaglandin-E ^ -methyl ester

A solution of 8.07 g of methyl-6-endo- (i, 2-dihydroxyheptyl) -3-oxo-bicyclo- / 3.1 -., 0 -) - he: can -? .- (2.2- dinethylheptanoate) (which was obtained according to Example 50) in TOO ml of pyridine v / ird stirred under nitrogen and cooled in an ice bath, while 10.0 ml of methanesilfonyl chloride are added dropwise over the course of about 15 minutes. The mixture is ^{stirred at 0} ° C. for 2.5 hours, then 5 ml of water are added dropwise, the temperature being kept below 5 ^° C. The mixture is diluted with 100 g of L'is and extracted with a dichloromethane-ether mixture (1: 1). The dichloromethane ether extract is mixed with ice-cold dilute salic acid (10Q ml conc. Hydrochloric acid, mixed with 400 ml ice and water), aqueous sodium bicarbonate and saturated aqueous

009827/2053 /

washed with sodium chloride, dried over sodium sulfate and evaporated under reduced pressure. This gives 10.2 g of oil, which is dissolved in 300 ml of acetone and stirred with 150 nil Water is diluted. The Genisch is left to stand at 25 ° C for about 20 hours. Then it is diluted with 300 ril of water, and concentrated under reduced pressure until most of the acetone is removed, and with a dichloroethane-ether solution (1: 3) extracted. The dichloromethane ether solution wia: d one after the other with dilute aqueous sodium carbonate solution and saturated aqueous sodium chloride solution • Washed, dried over sodium sulfate and reduced under Evaporated pressure. 10.0 g of 01 are obtained, which corresponds to 1300 g Silica gel is chromatographed, which in a Ethyl acetate-cyclohexane-Genisch (1: 1) is packed wet. The column is filled with 8.5 1 of an ethyl acetate-cyclohexrine mixture (2: 1), 2 1 10? S-igeru and 2.5 1 20 ^ -igem methanol in ethyl acetate eluted, collecting 100 nl fractions. Prik honing 84 to 106 are evaporated and combined to give 1.18 g of 1 5-epi-2,2-Dinethylprostaglandin-E ..- methylei; -ter; Mas- " senspck + rum + Jeaks at: 396, 376 and 360, IR absorption at 3420, 1730, 1320, 1250, 1195, 1150, 1075, 1025 and 970 cn ".

Fractions 116 to 130 v / are evaporated and combined, and No. ii receives 1.48 g of 2,2-Dinethylpros taglp.ndin-E ^ -nethylester, mass spectrum: peaks at 396, 378 and 360; IR absorption at 3390, 1730, 1320, 1250, 1195, 1150, 1075, 1020 and 970 cn " ¹ .

Example 52 2,2-dimethylprostaglandin-F ^ -ir.ethylester and

2,2-Dimethylprastaclpnri in F ^ o -ethyl ester

A solution of 100 ng of 2,2-dinethylpro5taglp-nriin- ±; _1- meTnjiester in 5 ml of isopropanol is cooled to 0 C in an ice bath, and a solution of 50 ng of tatrituriborohydride in Tnl V / water is added. The genic becomes in the melting

ORjQ JM _ÄL

009-827 / 2053

Stirred ice bath for 2.5 hours; Now the Realct I ons mixture is treated with 1 ml of acetone and stirred for 10 minutes. Diluted Acetic acid is added until the mixture is neutral, then it is concentrated under reduced pressure until the Most of the isopropanol and acetone has been removed. Of the The residue is diluted with 10 ril of water and washed with 15 ml of ethyl, acetate extracted. The ethyl acetate extract is over sodium sulfate dried and evaporated under reduced pressure; 100 mg of residue remain.

This procedure is carried out with 600 mg of 2,2-Dimethy! Prostaglandin-. E ..- methyl ester repeated as starting material, and 600 mg Raw product "are obtained. The two products were the same, as by thin-layer chromatographic analysis (silica gel), developed with ethyl acetate ;; Vanillic phosphoric acid reagent stains made visible, both products are combined (700 Eg) and over 70. g Eieseisäure'gel chromatographed, dae in an ethyl acetate-Oyclohexn.n mixture (2: 1) was packed wet. The column is filled with: 500 ml of ethyl acetate, 500 rel 1 ^ -igen, 500 ml · 3? 'I-igem " and; .500 ml of 10% methanol in ethyl acetate elrn'.Rrt, 25 ml fractions ^^ were taken. Eractions 32 - 34

D prostagland-in-2? ^^ -nethyl ester? Melting point: 54 - 60 ° C;

injected and combined, and 170 mg of 2,2-dimethyl prostagland-in-2? ^^ -nethyl ester? Melting point: 54 - 60 ° C; Mass spectrum peaks at 39S, 380, 362, 327 and 308.

Pralctions 51 and 65 are also evaporated and combined, and one receives 29O rag 2,2-Dinethylprpstaglandin-Piomethylester; Schnelzpunlct: 69-74 ⁰ Cj Magsenspektrumi peaks at 590, 380 _r 362, 327 and 308. \

Example 53. 2,2-dimethyl prostaglandin P ^ p \.

A solution of 200 mg 2,2-ester in-5 nil methanol is mixed with -2.8 ml 4.5 ^ -igera: aqueous potassium hydroxide, and the G-enisoh is about 20 hours

Stopped at 25 ⁰ G under nitrogen, thin-layer chromatographic analysis of the reaction mixture indicated that the reaction was complete. The mixture is diluted with 30 ml of water and extracted with 15 ml of ethyl acetate. The aqueous solution is acidified with cold, dilute hydrochloric acid and extracted with two 25 ml quantities of ethyl acetate. The ethyl acetate extracts are combined, washed three times with water, dried over sodium sulfate and evaporated. 182 mg of a crystalline residue are obtained, which is recrystallized from an ether-pentane mixture. 142 mg of 2,2-dimethylprostaglandin-Fo are obtained; Melting point: 102-106 ⁰ C; Mass spectrum 384, 366, '34S and 294.

Example 54 2,2-Dimethylprostaglandin-i ^l ₁₀ ,

Following the procedure of Example 53, but using 2,2-dimethylprostaglandin- ¹ ^ -methyl ester instead of 2,2-dimethylprostaglandin-]?., Β-methyl ester, 2,2-dimethylprostaglandin- is obtained. F | (^; melting point: 108 - 112 ⁰ C; mass

384, 366, 348 and 294. ·

Example 55 methyl 6-endo- (i-heptenyl) -3-oxo-bicyclo- J3, 1, Oj -hexan-2- (3,3-dimethylheptanoat)

A) 7-iodo-3j3-dimethylheptanoic acid methyl ester

A cold mixture of 110 ml of 96% sulfuric acid and 13 ml of water is stirred while 24 g of boron trifluoride gas are fed in. A mixture of 83 g of 6-chloro-2-methylhexan-2-ol and 107 g of 1 ", 1-dichloroethane is added to the sulfuric acid solution over a period of two hours while stirring vigorously ₅ v / above the temperature at 0-5 ° C. The mixture is then stirred for two hours ^{at 10-15 ° C. and poured over}

G ice-ego-seru The mixture v / lrd with a "Äther-- Skellyaolve-B mixture.. (1: 1) _s The extract with cold verdu.rin.ter · aqueous sodium hydroxide solution is extracted.

ϊ, ίΰΐΟ i; .Ar * - _T

This alkaline solution is acidified with dilute hydrochloric acid and extracted with an ether-Skellysolve-B mixture. The ether-Skellysolve B solution is washed with water, then with saturated aqueous sodium chloride, dried over sodium sulfate and evaporated under reduced pressure. 45 g of 7-chloro-3,3-dioethylheptanoic acid are obtained, which is dissolved in 125 ml of ether and to which an excess of diazomethane in ether is added at room temperature. After 3-5 minutes, the excess diazomethane is destroyed by adding acetic acid. The mixture is washed with dilute hydrochloric acid, dilute aqueous potassium hydroxide, water and saturated aqueous sodium chloride solution, dried over sodium sulfate and evaporated under reduced pressure. 48.6 g of methyl 7-chloro-3,3-dimethylheptanoate are obtained. A solution of 48.6 g of this ester in 750 ml of dry acetone and 75 g of sodium iodide are stirred for 40 hours with heating in a reflux condenser. The mixture is cooled and filtered and the filtrate is concentrated to remove most of the acetone. The concentrated filtrate is diluted with water and extracted with an ether-Skellysolve-B-Genisch (1; 1). The ether-ske lly solve B extract is washed with fresh, dilute aqueous sodium thiosulfate, water, and saturated aqueous sodium chloride, dried over sodium sulfate, and concentrated by evaporation under reduced pressure to give a residue. This residue is distilled, obtaining 61.5 g tIethyl-7-Dod - 3,3-dimethylheptanoat with a boiling point (mid-fraction) of 79 ⁰ C at 0.05 rnra. .

B) methyl - 6-endo- (1-heptenyl) -3-oxo-bieyclo - [_ 3,1, o7-nexane-2- (3,3-dimethylheptanoate)

Following the procedure of Example 49, but using methyl 7-iodo-3> 3-diinethylheptanoate instead of methyl 7-iodo-2,2-dimethylheptanoate, the compound methyl-6-endo- (1-h. eptenyl) -3-oxo-bicyclo -ß _} \ _f QJ- ,

009827/2053 BADORfQINAL

- 103 -

hexane-2- (3,3-dimethylheptanoate), which is chromatographically converted into the weaker polar and more polar isomer is decomposed.

Example 56 3.3 ~ ^e ~ Eim thylprbstaglandin e ^ methyl ester and

1 5 ^ -βρι-3,3-dimethylprostaglandin-S ^ --methylester ·,

Follow the procedure of Examples 50 and 51, but below Use of methyl-6-endo- (i-heptenyl) -3-oxo-bicyclü - ^ 3,1, O) -hexane-2- (3; 3-d ime thylheptanoat) (weaker ρ οlares Isomer) instead of the methyl-6-endo- (1-heptenyl) -3-oxo-bieyclo

1.0 | -hexane-2- (2,2-dimethylheptaiioate) In Example 50, the 3,3-bimethyl-prostaglandin-E .. -methyl ester of melting point 37-38 ⁰ j mass spectrum: 396, 578, 360, 325, 307 and 293; IR absorption at 3400, 1740, 13 ^ 5, 123p, 1150, 1130, -1075, 1015 and 965 cm "¹; and 15-epi-3,3-Ditvetl; yI-prostaglandin-E ^ -methyl ester; left spectrum: 396, 378, 360, 347, 346, 325, 307 and 293; IR absorption at 34 ^ 0, 1735, 1350, 1230, 1150-1135, 1075, 1015 and 970 ce " ¹ ,

Example 57 Diethyl-7- | endo-6- (1-heptenyl) -3-oxo-bicyclic- / 3,1, θ] -hex-2-ylj heptanoate »

A solution of 1.45 g of potassium tert-butoxide in 50 μl of tetrahydrofuran is added for 20 minutes while stirring . a solution of 1.00 g of Endo-6- (cis- and tr? Jip ' ^! -iieptey-yl) -bicycloTb, 1, Oj-hexan-3-one and 4.1 g of 7-J _f od-} ieFtaii {= aure ~ 5ietrylester in 25 ml tetrahydrofuran is added dropwise while nitrogen is bubbled through the reaction mixture. Then add 25 al 5% saline acid, the tetrahydrofuran is evaporated off, twice the volume of water is added, and the mixture is extracted three times with ethyl acetate. The . Combined extracts are washed with aqueous Hatriunthioeulfatlösung, .dry and evaporated .. The Eückstrind is cliromatpgraphiert on ϊ 00 · g silica gel, with 500 si Skellysolve B, 500 ml 2.55 ^ strength ethyl acetate in Skellysalve B, 1,500 ml .5> 5- igem ethyl acetate in.Skeilj ^ solve B and 700 sil

/ ^, ^: \ _; 009827/2 *

- ■ f

-. 104 ■ -

10 $ ethyl acetate is eluted in Skellysolve B, and 100 ml fractions are collected. Fractions 15-19 are combined and evaporated, and 366 mg of methyl 7- ^ endo - 6- (1 -heptenyl) -3-oxo-bicyclo- β , 1,07hex-2ov, .yl ~ y are obtained -hoptanoat. Fractions 20-24 are also pre-cleaned and filtered, and 151 ag of the corresponding 2β-yl-IDomG, i: 'Gu

Example 58 Ethyl-7- {endo-6- (1,2-dihydroxyheptyl7-3 / -oxo-

bicyclo- ^ J?, 1, <^ - hex-2 <Xj-yl7-heptanoate

A solution of 4.0 g of potassium chlorate and 0.2 g of osmine tetroxide in 48 ml of water is converted into a solution of 4 »0 g of methyl-7- εendo-6- (1 -heptenyl) -3-oxo-" bicyclo- / ?, 1, Oj-hez- ^ jrylf-teptanoat in 100 ml of tetrahydrofuran. The (Jocisch is ^{heated to 50 0} O for 5 hours with stirring. Then the tetrahydrofuran is evaporated, and 50 ml of water / water are added to the residue The mixture is extracted with three 150 μl quantities of dichloromethane. The combined extracts are washed with water, dried and evaporated. The residue is chromatographed on 400 g of silica gel with 4.4% ethyl acetate In Skellysolve B, 4 liters of 50% ethyl acetate in Skellysolve B and 1.2 liters of "ethylaceto.t" olulert, with 406 ml fractions being collected. Fractions 6-9 and fractions 1 2-14 are each combined and evaporated, and 1.47 and 1.18 g of the two isomeric forms are obtained, namely the weaker and the more polar form, respectively. Methyl-7- / endo-6- (1,2-dlhydroxyheptyl ) -3-oxo-bicyclo ~ £ 3.1.0 J- hex-2oi - ^ - yl7-heptanoate.

Example 59 Methyl ^ -Zendo-e- (1,2-dirriesyloxyheptyl) -3-

bicyclo- / 3,1, Oy-hex-2Äs-yl ^? -lieptanoate

Methanesulfonyl chloride (1 ml) is stirred into a solution of methyl-7- ^ endo-6- (1,2-dihydroxyheptyl) -3-OZo-DiCj ⁷ OIo- I, Λ , Oj-hex-iiXr-yl ^ - heptanoate (520 mg) in 4 ml of pyridine at ⁰ ° C. under a nitrogen atmosphere, the

# 09 8 27./ 2 Ό-5.3ν: ■. '·:'. ■■■ BAD ORIGINAL

19379-8

is " ^{stirred at 0} ° C. for two hours. Now 5 ml of ice-cold water are added and the mixture is stirred for 5 minutes. A mixture of ice and water (15 μl) is added and the entire mixture is mixed three times with 100 ml The combined extracts are washed successively with ice-cold aqueous sodium chloride solution, 10% sulfuric acid, the common salt solution, 10% aqueous sodium carbonate solution and the common salt solution and evaporated to give methyl-7- / endo-6 - (1, ■ 2 ^< -dimesyloxyheptyl) -3-oxo-bicyclic Q- £ ~ 3,1, oJ-hex ^ OC-ylJ-heptanoate. ■ .. ■■.

Example 6 0 PGE ^ -Methyl Ester

A solution; 1/6 of the dimesylate from Example 59 is kept for 16 hours at 25 ° G in a mixture of 4 ml of acetone and 2 ml of Vfesser. An equal volume of a saturated aqueous sodium chloride solution is now added and the acetone is evaporated off. The remaining solution is extracted with 80 ml of ethyl acetate. The extract is washed in succession with 10 ^ aqueous sodium carbonate solution and saturated aqueous sodium chloride solution, dried and concentrated; steamed. The residue is chromatographed on 10 g of silica gel and with 100 ml of 25%, 100 ml of 50%, 100 ml of 75% of ethyl acetate in Skellysolve B _for 100 ml of ethyl acetate and then with 100 ml 5 $ metheaiol eluted in ethyl acetate, collecting 20 ml portions. Precautions 13-16 are combined and evaporated to give 15.3 mg of 1S-epi-PGE ^ methyl ester. Fractions 17-20 are also combined and evaporated, and 14.9 g of PGE.-methyl ester are obtained.

Claims (1)

193790S 15 661 ΓΑΪ E II I AKS ϊ RÜ C HE Process for the preparation of a compound of the formula 0 < Ί5 IT in which a number from 1 to 8 and a is a number from 0 to 4, ^a | 5 is an alkyl radical with 1 to 4 carbon atoms, or a radical -CH ₂ CCl-, Ϊ = isobutyl _T tert-butylf 3,3-difluorobutyl -, 4,4-Üifluorobutyl or 4,4,4-2rifluorobutyl radical, unö '"^ ~ ⁱ indicates the bond of the G group in question to the ring in <Xj ~ or β-configuration, characterized in that one! ^ sulfonic acid ester of the formulaϊ
■■ -, σ '''' CH- (CiL,) -X Ij da R ₆ O ₂ SO ₂₆ in the η _? a, R ^ ^ and Y have the above meaning, R _{6 represents} an alkyl radical with 1 to 5 carbon atoms and ^ Mäie bond of the - (CH ₂ ) _n -COOR ^ ₅ ~ group to the Cyelopentanriug iniV- or ß-configuration as well shows exo- or enäo-lpriflguratiöii with regard to the gimppäenen on the cyclopropane Tiikg, at 0-60 ⁰ C with water inasetst *: ..- '■ "■ ■" - "..-..' 0 Γ! 9 ft 7 7 /? il ζ ^ ν!,. iJ U /. r> it U., J O -X- 2. Process for the preparation of a compound of the formula; H CHOH- (CH ₂ ) _a -Y in which η is a number from 1 to 8 and * a is a number from 0 to 4, R _{15 is} an alkyl radical with 1 to 4 carbon atoms, or -CH ₂ CCl-, Ϊ an isobutyl-, tert-butyl-, 3,3- Represent difluorobutyl, 4,4-mfluorobutyl or 4 _t 4,4-trifluorobutyl radical, and indicates the bond of the group in question to the ring in ° C or ß-configuration, characterized in that one disulfonic acid ester of the formula: ₆ O ₂ SO OSO ₂ R ₆ in the n, a, r.,. and X have the above meaning, R _{6 is} an alkyl radical having 1 to 5 carbon atoms and '^ the bond of the - (CH ₂ ) -COOR ₁ c group to the cyclopentane ring in <x.- or ß-configuration and with respect to the the cyclopropane ring-bound group indicates ezo or endo configuration, at 40-100 ⁰ C alt a combination of water, a base, which is characterized by a pH-Yert of the aqueous solution of 8-12, and a sur formation of a basic and essentially homogeneous reaction mixtures, sufficient quantity of an inert, water-soluble organic 009827/2053 ■ i:.;, -. ;;; ■ jot niche diluent converts, 3. A process for the preparation of a compound of the formula _n -COQH CHOH- (CH ₂ ) _a -Y in which η is a number from 0 to 8 and a is a count, from O to 4, Y is an isobutyl, tert-butyl, 3,3-difluorobutyl, 4 t represent 4-difluorobutyl or 4,4 »4-trifluorobutyl radicals, and '"" -' the attachment of the appropriate group to indicates the ring in <? C or ß configuration, characterized in that a compound of the Pormel HO ' _H > ^y CHOH- (CH ₂ ) _a ~ Y in which n, a, Y and / ^ have the above meaning, with zinc and an alkanecarboxylic acid "with 2 to 6 carbons converts material atoms. 4. Process for the preparation of a compound of the formula . ,. ^{0 = 0} \ ■■■■■ -. : ■ H ^ CHOH- (CH ₂ ) _a -Y. in which η is a number from 1 to 8 and a is a number from O to 4, Y is an isobutyl, fert-butyl, 009827/2053 4 »4-2ifluorobutyl- | or 4,4,4 -'- Crifluorobutyl radical, and ^/ " ^v ~ ^; indicates the bond of the corresponding group to the Hing inoC- or ß-configuration, characterized in that a compound of the formula: C = C C GHOH- (CH ₂ ) _a -Y C C in which n ', a, Y and - ^ have the above meaning, reacts with zinc and an alkanecarbonic acid with 2 to β carbon atoms. ¹ 5. Process for the preparation of a compound of the formula: (CH ₂ J _n - CHOH- (CH ₀ ) -Y in which η is a number from 1 to 8 and a is a number from 0 to 4,! Lg carpentry or an AXkyl radical with 1 to 4 carbon atoms, and ϊ an isobutyl, tert-butyl-j 3 ₉ 5-difluorobutyl, 4, Represent 4-difluorobutyl or 4 »4,4-sifluorobutyl radical and - ^ 'indicate the bond of the corresponding group to the ring in the X or S configuration, characterized in that a compound of the formula: ^H0 H ^ ^ CBOH- (CHq) -X 009327/2053 ■■■ :: ■ ^\: * -β -. :: ■ ... ■ λ ■ - ■ ..;. ■ .. in which R ^ g, η ₈ a »Y and '---' have the above meaning, dehydrated with an acid. 6. Process for the preparation of a compound of the formula HO Tin '^ «» «ν ην _H ^ ^ CHOH-- in which η is a number from 1 to 8 and a is a number from 0 to 4 »It. g of hydrogen or an alkyl radical having 1 to 4 carbon atoms and Y isobutyl-i tert-butyl, 3t3-difluorobutyl, 4,4-difluorobutyl - or 4,4,4-Trifluorobutylreet represent, and ^ the bond of the corresponding groups to the ring in (TO- or ß-configuration indicates, characterized in that a compound of the formula: CCHg) _n -COOH ₁₆ H H> GHOH- (CHg) ₃ -Y in which η, Lg ₁ a, I and '^ have the above meaning, converted with 6arbonyl group | reducing agent ^^. 7. Process for the preparation of a compound of the formulaί ■ '■■■' ■ ■■■■■■■ - ■■ '' ■■ - ■ - ^; "O, HO ": ■■ ^ GC \ _: Ή GHOH- (CH ₉ ) -Ϊ , in which sa'a smlil from 1 to 6 and a a number from 0 to A ₉ ■ '■ ■ "" · ■ Ö09827 / 2053 ■; "" .. ■ .. ". \ /""■■ R ₁ f. An alkyl radical with 1 to 4 carbon atoms or -CHpCCl-, 1 an isobutyl-, tert-butyl- _s 3 _f 3-difluorobutyl ~, 4,4-mfluorobutyl or 4 »4,4-irifluorobutyl radical, and Z a represented by 2 fluorine atoms, methyl or ethyl groups or by an alkyl radical with 3 to 4 carbon atoms substituted ethylene radical, and ^^ indicates the bond of the corresponding group to the ring ine * --- or ß-configuration, characterized in that a Dieülfensäureester the formula ⁰ «^ in which m, a, Ϊ, Z and R ^ have the above meaning, Rg represents an alkyl radical having 1 to 5 carbon atoms and - ^ - 'the bond of the - (CH ₂ ^ -Z-COOR ₁ , - group to the Cyclopentane ring indicates inco or fl configuration as well as exo or endo configuration with regard to the group attached to the cyclopropane ring, reacted with water at 0 - 60 ° C. · 8. Process for the preparation of a compound of formula i (CH ₂ ^ - in which m is a number from 1 to 6 and a is a number from 0 to 4, R ₁ S- is an alkyl radical with 1 to 4 carbon atoms, or -CH ₂ CCl ₃ , Y is an isobutyl, tert-butyl, 3, 3-difluorobutyl, 4,4-difluorobutyl or 4 »4» 4-2rifluorobutyl radical and Z is a 009827/20 53 by 1 or 2 fluorine atoms, methyl or ethyl groups or substituted by an alkyl group with 5 Ms 4 carbon atoms Represent ethylene radical, and ^^ the bond of group concerned to the ring ini ^ - or ß-conguration indicates, characterized in that a disulfenic acid ester of OiL-CH- (CH ₀ ) _β -Υ OO ^R 6 ° 2 ^SO,. in which ra, a, Y, Z and R ^ have the above meaning, R _{6 is} an alkyl radical with 1 to 5 carbon atoms and the - (CH ₂ ) _m ~ Z-C00R ^ ^ -G group to the cyclopentane ring iricC- or ß-configuration as well as exo- or endo-configuration with regard to the group attached to the cyclopropane ring, at 40-10O ⁰ G with a combination of water, a base, the aqueous solution of which has a pH of 3 to 12, and a to form a basic and substantially homogeneous reaction mixture, a sufficient amount of an inert, water-soluble organic diluent is reacted. 9. Process for the preparation of a compound of formula i , L-z-cooli H ΟΗ0Η- (0Η ₂ ) ₀ -Ύ ■■■■: ■ in "the ε a number from 1 to 6 and a a number from 0 to A ₁ Y ein.Isobutyl-, tert-butyl-, 3» 3-difluprbutyl-, 4,4-bifluorobutyl ■ or 4 _} 4,4 -2rlfluorobutyl radical, and Z is a by 1 or 2 fluorine atoms, methyl or ethyl groupexi or 009877/2053 V / * substituted by an alkyl group with 3 to 4 carbon atoms Represent ethylene residue, and ^^ - 'the bond the corresponding group to the ring incO or 0 configuration indicates, characterized in that a compound of the formula: -Z-COO-CH ₂ CCI ₅ CHOH- (CH ₂ ) ^ Y in which a, m, Y, Z and - ^^ have the above meaning, with zinc and an alkanecarboxylic acid having 2 to 6 carbon atoms. 10. Process for the preparation of a compound of the formula: (CH ₂ ) _m -Z-CQ0H G
^ CHOH- (CH ₂ ) _a -Y in which m is a number from 1 to 6 and a is a number from 0 to 4, Y is an isobutyl, tert _{-butyl- 9} 3,3-difluorobutyl, 4,4-mfluorobutyl or 4,4 "4-trifluorobutyl radical and Z represents an ethylene radical substituted by 1 or 2 fluorine atoms, methyl or ethyl groups or by an alkyl group having 3 to 4 carbon atoms, and- ^ the bond of the corresponding group to the ring inoO or ß-configuration, characterized _@ that one Connection of the mold. 009827/20 53 19379G8 (CH ₂ J ₁₀ - CHOH- (CH ₂ ) _a -Y in which a, ia, - Y, Z and '^' have the above meaning, reacts with zinc and an alkanecarbonic acid with 2 to 6 carbon atoms. 11 · Process for the preparation of a compound of the formula in which ^ m is a number from 1 to 6 and a is a number from 0 to 4, R-jg is hydrogen or an alkyl radical having 1 to 4 carbon atoms, Y is isobutyl, tert-butyl, 3 _{f is} 3-bifluorobutyl, 4 , 4-difluorobutyl or 4,4,4-trifluorobutyl radical and Z represent an ethylene radical substituted by 1 or 2 fluoro atoms, methyl or ethyl groups or by an alkyl group having 3 to 4 carbon atoms, and ^/ ^ ^/ the bond of the corresponding group to the Ring \ ixiO ^ - or ß-Kxmfiguration indicates, characterized in that a compound of the formula: ^H0 CHOH- (CH ₂ ) _a -Y in the a, m _s ϊ _? Z _g R ^ g and ^ s have the above meanings, dehydrated with acid. .- ■ ...--. . . . ■ ". -. 00 982 7/20 53 12. Process for the production of a compound of the molded ice HO (CH ₂ J _1n -Z-COOR ₁₆ H CHOH- (GH ₂ ) ^ - Y in which m is a number from 1 to 6 and a is a number from 0 to 4f R _{16 is} hydrogen or an alkyl radical with 1 to 4 carbon atoms, Y is an isobutyl, tert-butyl, 3 »3-difluorobutyl, 4 ₉ 4- Difluorobutyl or 4,4,4-trifluorobutyl radical and Z represents an ethylene radical substituted by 1 or 2 fluorine atoms, methyl or ethyl groups or by an alkyl group having 3 to 4 carbon atoms, and M the bond of the corresponding group to the ring in ^ - or ß-configuration , characterized in that a compound of the formula: in which a, m, Y, Z, R ₁₆ and "^ have the above meaning, is reacted with a carbonyl group reducing agent. 15. The method according to claim 1 or 7, characterized in that that the Dlsulfonsä ^ reestex · is in the exo configuration. 14. The method according to claim 1 or 7 »characterized in that that the disulfene acid ester is present in endo configuration. 15. The method according to any one of claims 1,7 »13 or 14, characterized known that Rg is a methyl group. 009827/2053 16. The method according to any one of claims 1, 7, l-3 _t H or 15 »dadiirch characterized in that sufficient amounts of an inert, water-soluble organic diluent are used to form a substantially homogeneous reaction mixture. 17 · The method according to claim 16, characterized in that acetone is used as a diluent. 18. The method according to claim 2 or 8, characterized in that that the base is a water-soluble salt of carbonic acid used. 19. The method according to claim 2 or 8, characterized in, that an alkali metal carbonate is used as the base 20. The method according to any one of claims 3 » A ₉ 9 or 10, characterized in that acetic acid is used as the alkanecarboxylic acid. 21. The method according to any one of claims 5 or 11, characterized 8 that acetic acid is used as the acid. • 22. The method according to claim 6 or 12, characterized in that that he. as a carbony group reducing agent hatriuiaborhydrid used. 23. The method according to any one of claims 1 to 22 _1, characterized in that the remainder - (CHp) ₁₁ -OOOR ₁ Jc or "CCHg) _1n -Z-COGR ,. c iniT ^ -Konfiguratioa is bound. 24. The method according to any one of claims 1 to 22, characterized in that the remainder - (CH ₂ J _n -COOR ₁ - or C is bound in ß- £ configuration. 0 09 8? -7 / 2.0.5.3 25. The method according to any one of claims 1 to 24 »characterized in that a means the number 1» if Y is difluorobutyl or trifluorobutyl, the number Z ₉ if Y is isobutyl, and the number 3 _β if Y is tert-butyl, 26. The method according to any one of claims 1 to 25 »characterized in that that Y is xsobutyl. 27. The method according to any one of claims 1 to 25, characterized in that Y is tert-butyl. 28. The method according to any one of claims 1 to 25, characterized in that Y is 3 »3-I> ifluerbutyl. 29. The method according to any one of claims 1 to 25 »characterized in that Y 4,4-difluorobutyJ. is. 30. The method according to any one of claims 1 to 25, characterized in, that Y is 4,4,4-trifluorobutyl. 31 »Method according to one of Claims 1 to 3O _9, characterized in that n, where _{s in} the present case means the number 6 · 32. The method according to any one of claims 1 to 3O _8, characterized in that _{where m s is} present, SaJiI 4 means 33 · Method according to one of claims 1 to 30 or 32 »characterized in that Z ₉ where _{9 is} a dsrch 1
Fluorine & tora substituted ethylene. 34 · Varfatesxi na @ ii one 'of claims 1 to 30 or 32, thereby g @ kennaeiohnet- "that Z ₉ where present" is followed by 2 is" 0.0S82.7 / 20 - 13 - 35 «Method n & eii one of claims 1 to 30 or 32 j characterized in that Z _8, where present, a through 1 Ethylenreet substituted by methyl group is 36. The method according to any one of claims 1 to 30 or 32, characterized in that Z, where present, a through 2 methf groups substituted on the same carbon atom Is ethylene radical. 37 · method according to one of claims 1 to 30 or 32, characterized in that Z, where present, a through an alkyl radical with 3 to 4 carbon atoms: which is is in £ <* position to the carboxyl function, substituted Is ethylene radical. 38, compounds of the formulas: II CHOH- (CH ₂ ) _a -Y CHOH- (CH ₂ ) _a ~ r or IXI to 4 _n -COOR _i3 .H η a% buoX rm. 1 to 8 and a a SaM. ? on0 hydrogen ^ a. Alkylreet isit 1 to 4 a pMsaa ^ ologiseh 009827 / 20B3 Represent 4,4-difluorobutyl or 4 _e 4,4-Trifluorbutylr © et ^^ and dle bond of the corresponding group to the ring - indicating or ß-conf iguration. 39. Connections of the formulas: HO II / T "( ^CH 2>xn" ^Z ~ ^COOR 13 HO ^ * a " ¹ III in which ra a number from 1 to 6 and a a number from 0 to 4 "Rj" hydrogen, an alkyl radical with 1 to 4 carbon atoms or a pharmacologically acceptable cation, Y an isobutyl, tert-butyl _e 3 ₉ 3 ~ i ) ifluorobutyl- _> 4 »4-difluorobutyl or 4,4 _f 4-trifluorobiatyl radical and Z represent an ethylene radical substituted by 1 or 2 fluorine atoms _s methyl or ethyl groups or by an alkyl group with 3 to 4 carbon atoms,« nd - ^ - the Bonding of the appropriate group to the ring In ^ - or ß-configuration anseigt. 40. A compound of the formula X of claim 38. 41. Compound of formula II of claim 38. 009827/2053 42. Connection of the pears! IIΓ of claim 38. 43. Compound of the formula I toil claim 39. 44. Compound of the formula II of claim 39. 45. Compound of formula III of claim 39 « 46. Connection according to one of claims 38 to 45, characterized in that the remainder - (CHp) -COOR., -, or - (CH ₂ ) J _n -Z-GOOR ₁₃ "± ϊϊ -X ^ -configuration bound is. 47. Compound according to one of claims 38 to 45 »characterized in that the remainder - (GH ₂ J _n -COOR ₁₃ or - (CII ₂ ) Z-COOR,. * Bound in ß-configuration lot. 48 «Connection according to one of claims 38 to 47, characterized characterized in that a is the number 1 when Y is bifluorobutyl or irifluorobutyl, the number 2 »when Y Isobutyl and the number 3 when Ϊ is tert-butyl. 49. Connection according to one of claims 38 to 48, characterized characterized in that Y is isobutyl. | - - ^ ■■, ■: ■ '■■■■. - ■■■ '. "' ■■ ■ - ■■■■■ '■■" ■ .-.,:''-' ■:. ■ "■ '- - 50. Connection according to one of claims 38 to 48, characterized characterized in that Y is tert-butyl. 51 * Connection according to one of claims 38 to 48, characterized marked ^. that --- Y is 3,3-mfluorobutyl. 52. Connection: according to one of claims 38 to 48, daduaieh Gelcerm shows that Y is 4,4-difluorobutyl. 53, connection according to any one of claims 38 to 48, characterized ; it is marked that - ■ ϊ ^^ 4,4,4 | - $ rifluorobutyl. 009827/2053 BATHROOM ORIGINATION η * 54. Connection according to one of claims 38 to 53, characterized characterized in that n, when * in the present case, means the number 6. 55. Connection according to one of claims 38 to 53 »thereby characterized in that m, when present, denotes the number 4. 56. Connection according to one of claims 38 to 53 or 55 »thereby characterized in that 2, if present, a through is a fluorine atom substituted ethylene radical. 57. A connection according to any one of claims 38 to 53 or 55 »characterized in that Z, if present, is a through two fluorine atoms on the same carbon atom more substituted Is ethylene radical. 58. Compound according to one of claims 38 to 53 or 55, characterized in that Z, if present, is a duroh is a methyl group substituted ethylene radical. 59. Compound according to one of claims 38 to 53 or 55, characterized in that Z, if present, a through zv / ei methyl groups on the same carbon atom is a substituted ethylene radical. 60. Compound according to one of claims 38 to 53 or 55 »characterized in that Z, if present, is an ethylene radical substituted by an alkyl group with 3 to 4 carbon atoms in the o <J position to the carboxyl group. For: The Upjohn Company HiAA) Attorney 00982 7/20