DK173287B1 - Phenylnonatetraenoic acid derivatives, a process for their preparation and pharmaceutical compositions containing them - Google Patents

Description

DK 173287 B1

The present invention relates to novel phenylnonatetraene acid compounds of the general formula I, R, R

r i7 i1 R ^ Λ .CH = CH-C * CH-CH = CH-C = CH-R, 987654321 1 R | represents hydrogen, chlorine, fluorine or trifluoromethyl; R 2 represents hydrogen, lower alkoxy, trifluoromethyl-lower alkoxy or hydroxy; R 4 represents straight chain length alkyl having 4-9 carbon atoms or - CH 2 (CH 2) + CH 2 OH; X represents -CH-0-. -CH-. -O- or -N-; R10 Rio R10 R10 10 Rs represents -C-0Ag;

ISLAND

and R7, R8, R9 and R1q independently represent lower alkyl or hydrogen; each n is an integer selected from 6 or 7; and salts thereof, wherein R 2 represents hydrogen, as well as pharmaceutical preparations containing them.

The compounds of general formula Z and their salts act as disease modifiers for the treatment of rheumatoid arthritis as well as related diseases such as osteoarthritis.

FIG. 1, 2, 3, 4, 5, G and 7 show schematic process steps for preparing the above compounds of general formula I.

The term "lower alkyl" as used herein refers to both straight and branched alkyl of 1-7 carbon atoms. Among the preferred lower alkyl groups are methyl, ethyl, isopropyl, n-butyl, etc., with methyl and ethyl being particularly preferred. The term "lower alkoxy" means lower alkoxy groups having 1-7 carbon atoms such as methoxy, ethoxy, isopropoxy, isobutoxy, etc. The term "trifluoromethyl-lower alkoxy" means a trifluoromethyl-substituted lower alkoxy substituent where lower alkoxy has the meaning given above.

2 DK 173287 B1

Also within the scope of the present invention are salts of the compound of the above general formula I with pharmaceutically acceptable, non-toxic, inorganic or organic bases, e.g., alkali metal and alkaline earth metal salt. Among the preferred salts are sodium, potassium, magnesium or calcium salts as well as salts with ammonia or appropriate non-toxic amines such as lower alkylamines, e.g., triethylamine, hydroxy-lower alkylamines, e.g., 2-hydroxyethylamine, bis- hydroxyethyl) amine or tris- (2-hydroxyethyl) amine, cycloalkylamines, e.g., dicyclohexylamine, or benzylamines, e.g., N, N * -dibenzylethylenediamine and dibenzylamine. These salts can be prepared by treating the compounds of general formula I wherein R 9 represents hydrogen, with inorganic or organic bases by conventional methods well known in the art.

The compounds of general formula I can be used to treat 15 patients suffering from rheumatoid arthritis or related diseases. In such cases, the compounds modify the effects of these diseases by reducing the destruction of the bone joints caused by this disease and reducing inflammation, heat and pain in the joints caused by rheumatoid arthritis and related diseases. The compounds of general formula I and their salts are also useful in the treatment of diseases due to immune hyperactivity such as transplant autoimmunity, autoimmune diseases, and graft versus host disease ("graft versus host disease"). The unexpected lack of toxicity of the compounds of the invention can be demonstrated by the compound (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid having a LD ^ value in mice exceeding 1,000 mg / kg both interperitoneally and orally.

3 DK 173287 B1

The compounds of the present invention are effective anti-arthritis agents which can be seen from the results obtained when these compounds are administered to rats in accordance with the chronic adjuvant arthritis testing system described by Billingham 5 and Davies, Handbook of Experimental Pharmacologists (eds. JR Vane and SH Ferreira), vol. 50/11, pp. 10B-144, Springer-Verlag, Berlin, 1979.

In this procedure, adjuvant arthritis was induced by sub-plantar injection on day 0 of 0.05 ml of adjuvant (a suspension of varicella, dried Mycobacterium butvricum, 0.5% (w / v), in heavy mineral oil containing 0.2 % digitonin) in Charles's right hind paw

Male River Lewis rats (120-140 g), which were placed individually in cages and fed ad libitum and water. Paw volumes (both hind legs) were measured immediately after injection of the adjuvant. Paw volumes were also measured to monitor the development of inflammation-induced swelling in the 15 arthritis paws at 3-7 day intervals by immersing the paw at the level of the lateral malleolus in a mercury plethysmograph.

The drugs were administered once a day (starting on the day of adjuvant injection) by incubation using Tween® 80 (polyoxyethylene sorbitan monooleate) at a dose of 0.25 ml / 100 g body weight as vehicle. Arthritis control rats received daily doses of carrier alone. On days 23-25, the rats were sacrificed, plasma was collected and plasma ibrinogen levels were determined (ammonium sulphate turbidimetric method), as described by Exner et al., Amer. J.

Cl in. Path. 71. 1979, pp. 521-527. The anti-inflammatory activity of the test drugs was determined by comparing the degree of paw swelling (paw volume on a particular day, i.e., day 4 to day 25, minus paw volume on day 0) of drug-treated arthritis rats with the degree of paw swelling in the carrier-treated arthritis rats. Drug-induced decreases in plasma fibrinogen levels, an acute phase 30 protein produced in plasma in rats with adjuvant-induced arthritis, were also used to quantify the anti-inflammatory activity.

The results of various compounds of the present invention in comparison with Indomethacin and vitamin 13 cis are listed in Table I below.

4 DK 173287 B1

TABLE I

»Reduction 1 Plis« * - ~ ''>

Or »l paw volume Mbrlene dosi * _pi day .3 ____ In out Increase in __STRUCTURE ___ μ Hol / tg Right Left (reduction] ___ bodyguard (g) _

Berer - ... ---- ---- 28 · J1 lndo * «lhicfn 3 -66.« 7 -10 t 46. 60 -31 -49 -18 -7 i

C02M

». <1 -S. -SS

1ν ^ Λ "00, Η".

TQL 7S - * 7 -? «-Σι * v (QT? 5 -5Ϊ -53 ♦ 24 75 -29 -32 · 35 ^ S ^ OCgHig ____________ 95 -37 -49 OS '» 0 (CH;) 8Om (Ol 75 In Table I above, the percent reduction in the paw volume shows the effectiveness of the compounds of the invention in reducing the swelling caused by adjuvant arthritis. The compounds of the invention 5 effectively reduce the swelling caused by the adjuvant, and the compounds of the invention were effective in reducing plasma fibrinogen, which is generally associated with rheumatoid arthritis.

Furthermore, it is apparent from the weight gain of the animals that the compounds of the invention at the dose tested did not produce any significant reduction in the weight gain of the animals. This shows the lack of toxicity of the compounds of the invention.

The compounds of general formula I and pharmaceutically acceptable salts thereof may be used for several pharmaceutical compositions. In these compositions, the compounds can be administered in the form of oral unit dosage forms such as tablets, pills, powders, capsules, and in such forms as injections, solutions, suppositories, emulsions, dispersions and in other convenient forms. The pharmaceutical compositions containing the compounds of general formula I are conveniently formed by admixing it with a non-toxic pharmaceutical organic carrier or a non-toxic pharmaceutical inorganic carrier. Typical pharmaceutically acceptable carriers are, for example, water, gelatin, lactose, starches, magnesium stearate, talc, vegetable oils, polyalkylene glycols, vaseline and other commonly used pharmaceutically acceptable carriers. The pharmaceutical compositions may also contain non-toxic excipients such as emulsifiers, preservatives and wetting agents and the like, such as, for example, sorbitan monolaurate, triethanol aminoleate, polyoxyethylene sorbitan, dioctyl sodium sulfosuccinate and the like.

The administered daily dose of the compounds, of course, depends on the particular compound being used, the route of administration chosen, and the size of the recipient. The dose administered is not dependent on specific limits, but it is usually in effective amounts according to the pharmacological function of the compounds of the invention. A representative typical method for administering the compounds of the general formula I or salts thereof is by oral administration. By this route, the compounds of the general formula I or their salts can be administered at a daily dose of from 0.5 mg / kg orally to 100 mg / kg orally. These compounds may preferably be administered daily to patients in oral unit dosage forms at daily doses of 1-30 mg / kg body weight, with doses of 1 to 10 mg / kg being particularly preferred.

The compound of general formula I wherein X represents -O- may

are prepared from compounds of the general formula III

Wherein R 1 and R 2 have the meaning given above, via the reaction scheme 5 shown in FIG. First

In the reaction scheme of FIG. 1, n, R 1, R 2, R 7 and R g have the meaning given above, and R 1 represents lower alkyl, Z represents a leaving group, Y represents aryl, preferably phenyl, and Z 1 represents halogen.

The compound of general formula III is converted to the compound of general formula IV via reaction step a) by reducing the aldehyde group to an alcohol. This reaction is carried out using a conventional reducing agent which converts aldehydes into alcohols.

Any conventional reducing agent in reaction step a) can be used for this purpose. In carrying out this reaction, it is generally preferred to use an alkali metal borohydride such as sodium borohydride as reducing agent. Any usual condition for performing reaction step a) may be used in such reduction reactions. If R 2 represents hydroxy, it is generally preferred to protect the hydroxy designated R 2 during the reduction 20 of the compound of general formula III and the subsequent conversion to the compound of general formula ΙΙ-Ά. Any conventional hydrolyzable hydroxy protecting group such as a lower alkanoyl group may be used to protect the hydroxy group when R 2 represents hydroxy. This ester protecting group can be cleaved by conventional ester hydrolysis after the formation of the Wittig salts of general formulas XIII and XVI or after the formation of the ether of general formula X.

The compound of general formula IV is converted to the compound of general formula VI via reaction step b) by treating compound 30 of general formula IV with a triarylphosphine hydrohalide. In this way, the phosphonium salt of the general formula VI is formed. Any conventional method of reacting an allyl alcohol with a triarylphosphine hydrohalide can be used to carry out this reaction. The phosphonium salt of general formula VI is reacted via a Wittig reaction with the compound of general formula VII in step c) to form the compound of general formula VIII. There. any of the conditions commonly used in Wittig reactions can be used to carry out reaction step c).

On the other hand, the compound of general formula III can be converted directly to the compound of general formula VIII via reaction with the phosphonium salt of the compound of general formula IX as in reaction step e). The reaction between the phosphonium salt of the general formula IX and the compound of the general formula III to form the compound of the general formula VIII is carried out using the same conditions described in connection with reaction step c).

The compound of general formula VIII is converted to the compound of general formula X by esterification or alkylation of the compound of general formula VIII with a compound of general formula V as in reaction step d). In the compound of the general formula V, Z can represent any conventional leaving group such as mesyloxy, tosyloxy or a halide. Any conventional method can be used to etherify a hydroxy group via reaction with a halide or leaving group to carry out reaction step d).

According to another embodiment of the invention, the compound of general formula X wherein the hydroxy group, when R 2 represents hydroxy, is protected via a hydrolyzable ester, can be prepared from the compound of general formula III by alkylation or etherification of the compound of the general formula formula III having the compound of general formula V to form the compound of general formula XI. This reaction is carried out by alkylating the compound of general formula III with the compound of general formula V as in step d). In reaction steps f) and d), where r4 represents hydroxyalkyl, hydroxy 30 contained in R4 need not be protected. This is because, under the conditions used in this reaction step, the compound of general formula V will react either with the compound of the general formula III or the compound of the general formula VIII to form the compound of the general formula XI or the compound of the general formula X , 35 without the need to protect the hydroxy group on the alkyl chain.

Alkylation or etherification will occur directly with the phenyl hydroxy moiety on either the compound of the general formula III or the compound of the general formula VIII, and there is only slightly, if at all, reaction with the hydroxy group on the alkyl chain represented by R4. The compound 40 of the general formula XI is converted to the compound of the general formula XII via reaction step g) by reduction. The same conditions described in connection with reaction step a) can be used to convert the compound of general formula XI to the compound of general formula XII.

e DK 173287 B1

The compound of general formula XII is converted via reaction step h) to the compound of general formula XIII by treating the compound of general formula XII with a triarylphosphine hydrohalide in the same manner as described above in step b). The compound of general formula XIII is converted to the compound of general formula X by reacting the compound of general formula XIII with the compound of general formula VII via reaction step i). This reaction step is carried out in the same manner as described above in connection with reaction step c).

According to another embodiment of the invention, the compound of the general formula X is prepared by first converting the compound of the general formula XI to the compound of the general formula XIV. The compound of general formula XI is converted to the compound of general formula XIV by aldol condensation with the compound of general formula XX. Any method of aldol condensation can be used to react the compound of the general formula XI with the compound of the general formula XX to form the compound of the general formula XIV. In the next step, the compound of general formula XIV is condensed via either a Grignard reaction with vinyl magnesium halide or a lithium condensation reaction with vinyl lithium to prepare the compound of general formula XV. Reaction step k) can be carried out using any of the conditions conventional for lithium condensations or Grignard condensation reactions. The compound of general formula XV is converted to the compound of general formula XVI by reacting the compound of general formula XV with a triarylphosphine hydrohalide in the manner described above in reaction step b) 30. The compound of general formula XVI is then converted to the compound of general formula X via reaction step m) by reaction with the compound of general formula XVII. Reaction step m) is performed using a Wittig standard reaction as described in connection with reaction step c). The compound of the general formula XVI by reaction with the compound of the general formula XVII gives the compound of the general formula X. The compound of the general formula X can be converted to the free acid, ie. the compound of general formula I, wherein R 5 represents COOH, by ester hydrolysis. Any conventional ester hydrolysis procedure will give the compound of general formula I wherein R 5 represents COOH.

The compound of the general formula I wherein X represents -N-, * 10

are prepared from compounds of general formula XXII

9 DK 173287 B1 where R | and R 2 has the meaning given above, via the reaction scheme shown in FIG. 2nd

5 In FIG. 2, R (, R 2, R 4, R 7, R 9, R 9 ', Z' and Y have the meaning given above. In Fig. 2, R 13 represents the same as R 4 with a carbon atom smaller than the alkyl group designated R 4. Therefore, R represents

X J

with 3-9 carbon atoms or -CH2 (CH2) mCH2OH, where m is an integer

less than n, i.e. m is an integer of 5-6. In FIG. 2 represents R

10 represents hydrogen or lower alkyl of 1-7 carbon atoms and R 'represents lower alkyl of 1-7 carbon atoms. In this embodiment, R "represents lower alkyl of one carbon atom less than the alkyl group represented by R".

10 In FIG. 2, the compound of general formula XXII is reacted with an acid chloride of general formula XIX, where Z 'represents halogen, to form the compound of general formula XXIII which is then converted either to the compound of general formula XXVI or to the compound of general formula XXXI. When R 3 in the compound of general formula XIX represents -0112- (012),, 0112011, where m is an integer 20 of 5-6 carbon atoms, the presence of the hydroxy group on the substituent R will not affect the reaction to form of the compound of the general formula XXVI or the compound of the general formula XXXI. It has been found that this hydroxy group remains unaffected throughout the series of reactions shown in FIG. 2nd

25 On the other hand, this hydroxy group can be protected by forming a hydrolyzable functional ether group. which protects the hydroxy group during these reactions. Any conventional ether protecting group may be used to protect the hydroxy group during these reactions. Among the preferred ether protecting groups are: tetrahydropyranyloxy, tert-butoxy, tri-lower alkylsilyloxy, i.e. trimethylsilyloxy, etc. Any conventional ether protecting group may be used to protect the terminal hydroxy group which may be present as R. On the other hand, in FIG. 2 reactions are performed without any protection of the terminal hydroxy group.

In the first step of the reaction, the compound of the general formula XXII is reacted with the compound of the general formula XIX to form the compound of the general formula XXIII. Any conventional method of condensing an amine with an acid halide can be used to carry out this reaction. In the next step, the compound of general formula XXIII is converted to the compound of general formula XXIV via reaction step n) by treating the compound of general formula XXIII with a reducing agent. Any conventional alkali metal aluminum hydride reducing agent may be used to perform this reaction, the preferred reducing agent being lithium aluminum hydride. Any of the conditions conventional for reduction with an alkali metal aluminum hydride reducing agent may be used to carry out this reaction.

The compound of general formula XXIV can be converted to compound 15 of general formula XXVI via intermediate XXV. In the first step of this process, step o), the compound of general formula XXIV is converted to the phosphonium salt of general formula XXV by reaction with a triarylphosphine hydrohalide as described above in connection with reaction step b). The compound of general formula 20 XXV is converted to the compound of general formula XXVI via reaction step p) by reaction with the aldehyde of general formula VII (cf. Fig. 1). Reaction step p) to prepare the compound of general formula XXVI is carried out by a Wittig reaction in the same manner as described in reaction step c) above. The compound of the general formula XXVI wherein R 9 represents lower alkyl may, if desired, be converted to the free acid by conventional basic hydrolysis. Any basic hydrolysis method can be used to hydrolyze ester to convert the compound of general formula XXVI to the free acid. On the other hand, the compound of general formula XXVI wherein R4 contains a terminal hydroxy group etherified with a conventional ether protecting group can be converted to the free alcohol by acid hydrolysis. Any of the conventional methods of hydrolysis of ethers can be used to carry out this reaction. The ether hydrolysis of the compound of the general formula XXVI can be carried out either before or after the acidic hydrolysis used to hydrolyze the ester protection Ry '. On the other hand, the compound of the general formula X, if R4 in the compound of the general formula X of FIG. 1 contains an etherified hydroxy group, hydrolyzed in the same manner as the compound of the general formula XXVI.

On the other hand, the compound of general formula XXIV can be converted to the tertiary amine compound of general formula XXXI. In this reaction, the compound of general formula XXIV is reacted via reaction step q) with the acid halide of general formula XIX-A to prepare the compound of general formula XXVII in the same manner as described above in connection with the reaction for conversion. of the compound of general formula XXII to the compound of general formula XXIII.

The compound of general formula XXVII is converted to the compound of general formula XXIX via reaction step r) by treating the compound of general formula XXVII with a lithium aluminum hydride reducing agent as described in connection with reaction step n).

In the next step of this reaction scheme, the compound of general formula XXIX is converted to the compound of general formula XXX via reaction step s) by treatment with a triarylphosphine hydrohalide.

This reaction is carried out in the same manner as described in connection with reaction step b) above. The compound of general formula XXX 15 is converted to the compound of general formula XXXI via reaction step t) by reaction with the compound of general formula VII {FIG. 1).

In carrying out reaction step t), a Wittig reaction is used. Reaction step t) can be carried out in the same manner as described above in connection with reaction step c). The compound of general formula XXXI, wherein R 9 'represents lower alkyl, may, if desired, be converted to the corresponding compound of general formula XXXI containing the free acid group by basic hydrolysis. On the other hand, the compound of general formula XXXI, if R4 contains a terminal hydroxy group protected by formation of a hydrolysable ether, can be converted to the corresponding compound, where R4 represents a free hydroxy group, by conventional ether hydrolysis. This ether hydrolysis can be carried out before or after hydrolysis of the ester group designated R9 '.

In the reaction scheme of FIG. 2, when R 2 represents hydroxy, it is preferred that this hydroxy group be protected via formation of an ester protecting group. The ester protecting group may be removed after formation of the wittig salts of general formula XXX.

Those compounds of general formula I wherein X represents -CH-,

Rio

35 is prepared from a compound of general formula XXXV

X 1 represents Z 2 represents ether, bromine or iodine, R 1 and R 2 have the meaning given above and R represents hydrogen or lower alkyl as shown in Fig. 3. Figure 3 has Rj, R2, R4, R7, Rg, R'9, R1Q, R, Y, Z 'and Z "as defined above.

5 In FIG. 3, the compound of general formula XXXV is first reacted with the compound of general formula XXXIV via reaction step u) to prepare the compound of general formula XXXVI. This reaction is carried out via a Wittig reaction. In the compound of the general formula XXXVI, R may, if desired, represent -CH2- (CH2) m-OH, where the free hydroxy group may, if desired, be protected by forming any of the above-mentioned conventional ether groups. On the other hand, it has been found that this OH group can be a free hydroxy group and need not be protected by an ether protecting group. In carrying out the reactions of FIG. 3, this free hydroxy group is not affected by the reactions which convert the compound of general formula XXXVI to the compound of general formula ΧΧΧΧΓ. However, in order to obtain the best yields, it is generally preferred to protect this hydroxy group via formation of a hydrolyzable ether.

Reaction step u) is performed via a Wittig reaction between the compound of general formula XXXV and the compound of general formula XXXIV using the same reaction conditions described above in connection with reaction step c).

The compound of the general formula XXXVI can be converted to the compound 25 of the general formula XXXVII via reaction step v) by hydrogenation.

Any hydrogenation method can be used to carry out this reaction. The conventional hydrogenation methods include treatment of the compound of general formula XXXVI in an inert organic solvent medium with hydrogen gas in the presence of a catalyst. Any conventional hydrogenation catalyst may be used in carrying out this reaction. Among the preferred catalysts is palladium. In carrying out this reaction, any conventional inert organic solvent may be used. Furthermore, in reaction step v) any of the conditions conventional for catalytic hydrogenation can be used.

In the next step of this reaction, the compound of the general formula XXXVTI is converted to the compound of the general formula XXXIX via reaction step w) by treating the compound of the general formula 40 XXXVII with formaldehyde or a formaldehyde-releasing compound. In carrying out this reaction, the compound of the general formula XXXVII is first metallized with alkali metal alkyl, e.g. n-butyllithium. This reaction is generally carried out in an inert organic solvent such as an ether solvent. Among the preferred solvents are diethyl ether and tetrahydrofuran. In carrying out this reaction, the temperature and pressure are not critical. The reaction can be carried out at room temperature and atmospheric pressure. If desired, higher and lower temperatures can be used. After treating the compound with. the general formula XXXVIII with alkali metal alkyl is added formaldehyde or a formaldehyde-releasing compound to the reaction medium. Any compound capable of delivering formaldehyde such as paraformaldehyde can be used in carrying out this reaction. The reaction is carried out in the same reaction medium and using the same conditions under which the metallization of the compound of general formula XXXVIII was carried out.

The compound of general formula XXXIX is converted to the compound of general formula XXXX via reaction step x) by treating the compound of general formula XXXIX with triarylphosphine hydrohalide. This reaction is carried out in the same manner as described in connection with reaction step b) above. The compound of general formula XXXX 20 is converted via the reaction step y) to the compound of general formula XXXXI by reaction with the compound of general formula VII (cf. Fig. 1). Reaction step y) is carried out via a Hittig reaction using the same conditions as described in connection with reaction step c). The compound of general formula XXXXI can be converted to the corresponding compound containing the free carboxyl group instead of R9 '. This reaction is carried out by conventional ester hydrolysis in the manner described above. Any conventional ester hydrolysis process can be used. If R4 contains a terminal hydroxy group protected by use of an ether protecting group, that ether group can be hydrolyzed, giving the free hydroxy group by conventional ether hydrolysis such as using an aqueous inorganic acid. Any conventional method of ether hydrolysis can be used. The protected ether hydroxy group can be hydrolyzed either before or after hydrolysis of the ester group to form the free acid of the compound of general formula XXXXI.

If it is desired to prepare compounds of the general formula in which X represents -C = CH-, the compound is reacted with the general formula R10 40 flour XXXV of FIG. 3 via reaction step u) with the compound of general formula XXXIV, where R = R4, to form the compound of general formula XXXVI where = R4. This compound of general formula XXXVI, where R4 = R4, is then subjected to the same series of reactions as the compound of general formula XXXVII, ie. reaction steps w), x) and y), to form the compound of general formula I wherein X represents - ^ = CH-.

R10 In the reaction scheme of FIG. 3, where R 2 in the compound of the general formula XXXV represents hydroxy, it is preferred to protect this hydroxy group via esterification with a lower alkanoic acid. This ester protecting group may be cleaved after formation of the Wittig salt of the general formula XXXX.

The compound of general formula I and II wherein X represents -CH-O, * 10

may be prepared from a compound of the general formula L

VfHZ \ φζ * 1 wherein R 1, R 2 and Z "have the meaning given above, via the reaction scheme of Fig. 4, In Fig. 4, R (, R 2, R 7, R 8, R 10 * R 9 ', Y and Z "has the meaning given above and R = R 4, wherein a hydroxyl group contained in R is protected in the form of a hydrolysable ether group such as tetrahydropyranyl and the ether groups mentioned above.

The compound of general formula L is converted to a compound of general formula LI by reaction with an alkali metal alkoxide of general formula L-A. This reaction is carried out by reacting the compound of general formula L with the compound of general formula L-A using the conventional conditions for reacting an alkali metal alkoxide with a halide.

In this step, the compound of the general formula LI is converted to a compound of the general formula LII by first treating the compound of the general formula LI with an alkyl lithium such as n-butyl lithium to metallize the compound of the general formula LI. The metallized compound of the general formula LI is then reacted with 30 formaldehyde or a formaldehyde-releasing compound. In the conversion of the compound of general formula LI to the compound of general formula LII, the same reaction conditions as described in connection with reaction step w) are used. The compound of the general formula LII is converted to a phosphonium salt of the general formula LUI, the compound of the general formula LII being treated with a triarylphosphine hydrohalide in the manner indicated in reaction step b) above. The phosphonium salt of the general formula Lill is reacted via a Wittig reaction with the compound of the general formula VII (cf. Fig. 1) to form a compound of the general formula LIV. The reaction to form the compound of the general formula LIV is carried out in the same manner as described in step c) above.

When R in the compound of the general formula LIV contains a protected hydroxy substituent, this substituent can be hydrolyzed to form the free hydroxy compound by conventional methods of hydrolysis of ether groups which can be readily removed. Any conventional method can be used to hydrolyze ether protecting groups. The conditions conventional for the hydrolysis of ether protecting groups will not affect the other ether group in association with the general formula LIV. The compound of the general formula LIV can be converted to the free acid by conventional ester hydrolysis.

If R 2 in the compounds of the general formulas L, LI, LII and LUI represents hydroxy, it is preferred that the hydroxy group is protected via a hydrolyzable ester group such as lower alkanoyloxy. The hydrolyzable ester protecting group may be cleaved after formation of the Wittig salt of the general formula LUI.

If desired, the double bonds of the compound of general formula I at positions 2-3, 4-5, 6-7 and 8-9 can be either in cis or in trans configuration. On the other hand, these compounds may be a mixture of the various cis and trans isomers. In the compound of general formula VII, the double bonds therein may be either in cis cells in trans configuration depending upon the desired stereoconfiguration of the double bonds in the compounds of general formula I. The Wittig reaction carried out in the preparation of compounds 30 of general formulas I and II, as in steps c), e), i), etc., gives the double bond at the 8-9 position as a mixture of 8-9 cis and trans isomers. These cis and trans isomers can be cleaved by conventional methods such as fractional crystallization, etc.

In addition, when the compounds of general formula I have a double bond in the trans configuration at the 2-3 position, this isomer can be converted to the corresponding cis double bond by conventional isomerization methods known in the art. These methods include: treating the compound of general formula I with iodine in an inert organic solvent. Isomerization with iodine 40 gives the compound of general formula I with a 2-3 double bond at the cis position.

16 DK 173287 B1

The compounds of general formula I include all of the geometric isomers, including mixtures of these geometric isomers.

The compound of general formula XI wherein R 1 represents fluorine is a novel compound and can be prepared from a compound of general formula LV

"1" rk.

* 3 wherein R 2 has the meaning given above via the reaction scheme of FIG.

5. In FIG. 5 has R 2 and the meaning given above. R 3 is hydrogen.

In FIG. 5, the compound of the general formula LV is alkylated by reaction 10 with an allyl bromide. If the compound wherein R 2 represents hydroxy is desired, then, as a starting material, the compound of general formula LV wherein the hydroxy group designated R 2 is protected by esterification, i.e. the compound of the general formula LV wherein R 2 represents a protected hydroxy group. Any conventional method of alkylating a hydroxy group with an allyl bromide can be used to carry out the reaction of converting the compound of the general formula LV into the compound of the general formula LVI. The compound of general formula LVI is rearranged to a compound of general formula LVII by heating the compound of general formula LVI to a temperature of 190-230 ° C.

This rearrangement can take place without the use of any solvent or in the presence of a high boiling point hydrocarbon solvent. When R 3 represents hydrogen, the compound of general formula LVII is formed as a mixture with the isomer of the compound of general formula LVII where the allyl group sits para to the fluorine substituent on the benzyl ring. This isomer can be cleaved or used in subsequent reactions and separated from the reaction mixture in a later step.

The compound of general formula LVII is then converted to a compound of general formula LVIII by reaction with the compound of general formula V (Figure 1) as described in reaction step d) above. In the next step of the reaction scheme, the compound of general formula LVIII is converted to a compound of general formula LIX by isomerization with a strong base such as an alkali metal alkoxide in the vicinity of an inert organic solvent, preferably potassium tert.butoxide in dimethyl sulfoxide. The compound of the general formula LIX is converted to a compound of the general formula LX by treating the compound of the general formula LIX with ozone gas. In carrying out this reaction, temperatures of -70 ° C to -20 ° C are used. This reaction is further carried out in an inert organic solvent. Any conventional inert organic solvent may be used, preferably halogenated hydrocarbons such as methylene chloride.

A compound of general formula LX is the compound of general formula XI, wherein R (represents fluorine. This compound can be converted to a compound of general formula X according to the reaction scheme of FIG.

First

When R 2 represents hydroxy in the compound of general formula III, there are two hydroxy groups. Therefore, it is generally preferred to prepare the compound of general formula XI wherein R 2 represents protected hydroxy from a compound of general formula LXI as shown in FIG. 6. In this way, compounds of the general formula I, wherein X is -O-, R2, is hydroxy, and R4 is as defined above. In FIG. 6, R 1 and R together with the attached oxygen atom represent hydroxy protected with a conventional hydrolyzable protecting group, preferably lower alkanoyl.

In FIG. 6, the compound of the general formula LXI is converted to a compound of the general formula LXII using the same reaction as described in connection with the conversion of a compound of the general formula LV to a compound of the general formula LVI (cf. FIG. 8). The compound of the general formula LXII is then converted to a compound of the general formula LXIII using the same procedure as described in connection with the conversion of the compound of the general formula LVI to the compound of the general formula LVI I. In the next step, the compound is converted. with the general formula LXIII for a compound of the general formula LXIV by the same procedure as described in connection with step g ') of FIG. 5 and then to a compound of general formula LXV by the method described in connection with step h ') of FIG. 5. The conversion of the bromobenzene compound of the general formula LXV to the phenol compound of the general formula LXVI takes place by well known methods such as those described, for example, by Kidwell and Darling, Tetrahedron Letters.

1966, pp. 531-535.

In the next step for the preparation of the intermediate of the general formula XI wherein R 2 represents protected hydroxy, i. compound of the general formula XI-A, the hydroxy group of the compound of the general formula LXVi is protected by esterification with any conventional hydrolyzable ester group to form a compound of the general formula LXVII where R together with the bond oxygen 5 forms a hydrolyzable ester group. Any conventional method can be used to esterify a hydroxy group with an organic acid such as lower alkanoic acid having 1-7 carbon atoms to prepare the compound of general formula LXVII. The compound of the general formula XI * A is formed from the compound of the general formula LXVII in the reaction described above in connection with the conversion of a compound of the general formula LIX into a compound of the general formula LX (cf. Fig. 5). .

In converting a compound of general formula XI-A into a compound of general formula XVII as shown in FIG. 1, it is generally preferred to hydrolyze the ester substituent constituting R 1 after the formation of the Wittig salt of general formula XIII or XVI.

According to another embodiment of the invention, the compound of general formula XI, wherein Rf represents CF3 (the compound of general formula XI-B), can be formed by the one of FIG. 7 from a compound of the general formula LXX. In the first step of this reaction, the compound of the general formula LXX is converted to a compound of the general formula LXXI using the same procedure as described above in the reaction of step d) wherein the compound of the general formula VIII is reacted with a compound 25 of the general formula V to form a compound of the general formula X. In this reaction, where R2 represents OH, alkylation occurs very slowly on the hydroxy group sitting ortho relative to the CF3 group. Therefore, protection of this group may not be necessary since the alkylation preferably occurs with the meta-hydroxy group. Any 30 mixtures of alkylated products obtained by this reaction can be separated by conventional separation methods. The compound of the general formula LXXI is converted to the compound of the general formula XII-B by conventional methods for formylating a benzene ring such as by treatment with alkyl lithium and dimethylformamide.

The invention is further illustrated by the following. In the examples, the ether is diethyl ether and the solvents were removed under vacuum.

EXAMPLE 1 [[{2- (Nonyloxy) phenyl] methyl] triphenylphosphonium bromide 110 g of 2-hydroxybenzaldehyde was alkylated by mixing this compound with 180 g of 1-bromnonane, anhydrous potassium carbonate and 800 ml of dimethylformamide. This mixture became. heated at 80 ° C for 14 hours.

Hexane and water were then added and the hexane extract was concentrated and the residue was distilled to give 210 g of 2-nonyl oxybenzaldehyde, boiling point 121 ° C (0.3 mm Hg). A solution of 100 g of 2-nonyloxybenzaldehyde prepared as described above in 1000 ml of 10 ethanol at 10 ° C was reduced by treatment with an excess of 6 g of sodium borohydride, and after stirring the mixture for an additional 15-20 minutes at room temperature, the compound 2 nonyloxybenzyl alcohol isolated by extraction in hexane. Removal of the hexane in vacuo gave 98 g of crude 2-nonyloxybenzyl alcohol. The resulting 2-nonyloxybenzyl alcohol was added to a mixture of 144 g of triphenylphosphine hydrobromide in 500 ml of acetonitrile, and the resulting solution was refluxed for 14 hours. Removal of the solvents in vacuo and crystallization of the residue of a tetrahydrofuran / ethyl ether mixture gave 208 g of pure [[(2- (nonyloxy) phenyl] methyl] triphenylphosphonium bromide.

Example 2 [(2-Hydroxyphenyl) methyl] triphenylphosphonium bromide 2-Hydroxybenzyl alcohol was treated with triphenylphosphine hydrobromide in acetonitrile as described in Example 1 to give 25 [(2-hydroxyphenyl) methyl] triphenylphosphonium bromide.

EXAMPLE 3 (All-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester

A solution of 1 mole of [(2-hydroxyphenyl) methyl] triphenylphosphonium bromide in tetrahydrofuran was converted to the ylide at -35 ° C with a solution of n-butyllithium in hexane (2.1 mole equivalents) and then exposed to mole 7 -formyl-3-methyl-2,4,6-octatric acid ethyl ester and then stirred at -70 ° C for a further 15 minutes. Isolation of the organic products with a hexane / ethyl acetate mixture (4: 1 parts by volume) and dilute mineral acid (2M aqueous HCl) gave the pure (all-9-(2-hydroxyphenyl) -3), 7-Dimethyl-2, 4,6,8-nonatetraenoic acid ethyl ester (90% yield) after chromatography followed by crystallization of a dichloromethane / hexane mixture.

Example 4 (All-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester

A mixture of 0.5 mole of 2-hydroxybenzaldehyde and 0.6 mole (7-carboxy *).

2,6-Dimethyl-2,4,6-heptatrien-1-yl) triphenylphosphonium bromide in 750 ml of 1,2-epoxybutane was refluxed for 30 minutes, cooled, poured into an ether / hexane mixture (1: 1 parts after volume), filtered and concentrated. The residue was then crystallized by a hexane / ether mixture to give (all-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester (38% yield), m.p.

14 -14 5 ° C.

Example 5 (All-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid

A solution of 150 g of [[2- (nonyloxy) phenyl] methyl] triphenylphosphonium bromide in 1100 ml of tetrahydrofuran was cooled to -50 ° C to give a fine suspension of the solid salt, to which was added 20 180 ml of a 1 6M solution of n-butyllithium in hexane to give a solution of the ylide The mixture was then stirred for a further 15 minutes at -40 ° C, cooled to -70 ° C and treated with 65 g of 7-formyl-3-methyl-2 4,6-Octatrienoic acid ethyl ester dissolved in 250 ml of tetrahydrofuran Adding hexane and 40% aqueous methanol to the reaction mixture followed by concentration of the hexane extract gave 64 g of (all-E) -9- [2- (nonyloxy) phenyl ] -3,7-Dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester (58% yield), mp 52-53 ° C. This ester was then hydrolyzed to give a solution of 70 g of the ester in 1000 ml of ethanol. This solution was treated with 80 g of aqueous potassium hydroxide (in 400 ml of water) and heated under reflux for 1 hour. Water and aqueous mineral acid was then added and the solids extracted in chloroform. Concentration of this organic extract and crystallization of the residue of an ethyl acetate / hexane mixture gave 38 g of (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8- nonatetraenoic acid, mp 102-103 ° C.

EXAMPLE 6 21 (All-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid

A mixture of 24 g of sodium hydride (50% by weight in mineral oil) and 1000 ml of dimethyl formamide at 10 ° C was treated with 0.4 equivalents of 5 (all-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2.4 , 6,8-nonatetraenoic acid ethyl ester. The resulting mixture was then stirred at room temperature until al. hydrogen evolution was stopped to form the sodium salt of (all-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester. A solution of 0.5 equivalents of 1-nonyl-10 tosylate in 200 ml of dimethylformamide was then added to this brine, and the reaction mixture was stirred at 45 ° C for 14 hours.

Hexane / water was then gently added, the hexane extract was concentrated and the residue was purified by chromatography on silica gel. Crystallization of hexane then gave the pure (all-E) -9- [2- (nonyl-oxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester. Hydrolysis of this ester as in Example 5 gave (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid.

EXAMPLE 7 (All-E) - 3,7-dimethyl-9- [2 - [(2,2-dimethyloctyl) oxy] phenyl] -2,4,5,8-20 nonatetraenoic acid 2-Hydroxybenzaldehyde was condensed with 2, 2-dimethyl-1-iodooctane to give 2- (2,2-dimethyloctyloxy) benzaldehyde which was reduced to 2- (2,2-dimethyloctyloxy) benzyl alcohol and then converted to ([2- (2,2-dimethyloctyloxy)) phenyl] methyl] triphenylphosphonium bromide as in Example 1. Condensation of this phosphonium bromide with 7-formyl-3-methyl-2,4,6-octatric acid ethyl ester as described in Example 3 followed by hydrolysis as in Example 5 gave (all-E) - 3,7-dimethyl-9- (2 - [(2,2-dimethyloctyl) oxylphenyl] -2,4,5,8-nonatetraenoic acid, m.p. 113-117 ° C (of a dichloromethane / hexane mixture).

Example 8 (All-E) -3,7-Dimethyl-9- [2 - [(octyloxy) methyl] phenyl] -2,4,6,8-nonatetraenoic acid

Lithium octanoate prepared from octanol and n-butyllithium in a mixture of tetrahydrofuran / hexane and dimethylformamide was condensed with 2-bromobenzyl bromide to give 2- (octyloxy) methyl bromo-benzene. This material was treated with n-butyllithium in an ether / hexane mixture and then treated with paraformaldehyde to give 2- (octyloxy) methylbenzyl alcohol. This material was then treated with triphenylphosphonium bromide to give 5 [(2 - [(octyloxy) methyl] phenyl] methyl] triphenylphosphonium bromide.

Condensation of this material with 7-formyl-3-methyl-2,4,6-octatric acid ethyl ester as in Example 3 followed by hydrolysis as in Example 5 gave (all-E) -3,7-dimethyl-9- [2 - [(octyloxy) methylphenyl) -2,4,6, B-nonatetraenoic acid, m.p. 120-121 ° C (of a dichloromethane / hexane mixture).

EXAMPLE 9 (All-E) -9- [2-Chloro-6- (nonyloxy) phenyl] -3,7-dimethyl-2,4, S, B-nonatetraenoic acid 2-Chloro-6-hydroxybenzaldehyde was alkylated with 1- bromnonane as in Example 1 to give 2-chloro-6-nonyloxybenzaldehyde. Reduction with sodium borohydride as in Example 1 gave 2-chloro-6-nonyloxybenzyl alcohol, which after treatment with triphenylphosphine hydrobromide in acetonitrile as in Example 1 gave [[2-chloro-6-nonyloxy] phenyl] methyl] triphenylphosphonium bromide. Condensation with 7-formyl-3-methyl-2,4,6-oc-tetraenoic acid ethyl ester as described in Example 3 gave (all-E) -9- (2-chloro-6- (nonyloxy) phenyl) -3.7 dimethyl-2,4,6,8-nonatetraensyreet-ethyl ester. The ester was subjected to hydrolysis as in Example 5 to give (all-E) -9- [2-chloro-6- (nonyloxy) phenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, m.p. 129 -131 ° (of an ethyl acetate / hexane mixture).

EXAMPLE 10 (All-E) -9- (5-methoxy-2-nonyloxyphenyl) -3,7-dimethyl-2,4,6,8-nona-tetraenoic acid 5-Methoxy-2-hydroxybenzaldehyde was alkylated with nonyl bromide and reduced with sodium borohydride as in Example 1 to give 5-methoxy-2-nonyloxybenzyl alcohol, which upon exposure to triphenylphosphine hydrobromide gave [(5-methoxy-2-nonyloxyphenyl) methyl] triphenylphosphonium bromide. Condensation of this material with 7-formyl-3-methyl-2,4,6-octatric acid ethyl ester as in Example 3 followed by hydrolysis as in Example 5 gave (all-E) - 9- (5-methoxy-2-nonyloxyphenyl) - 3,7-dimethyl-2,4,6, B-nonatetraenoic acid, m.p. 125-126 ° C (of methanol).

EXAMPLE 11 (All-E) - 9- [2- (8-hydroxyoctyl) oxy] phenyl-3,7-dimethyl-2,4,6,8-nona-tetraenoic acid

The sodium salt of (all-E) -9- (2-hydroxyphenyl) -3,7-dimethyl-2,4,6,8-5 nonatetraenoic acid ethyl ester in dimethylformamide prepared as described in Example 6 was treated with 1,8-dihydroxyoctane monotosylate as described in Example 6 to give (all-E) -9- (2- (8-hydroxyoctyl) oxy] phenyl-3,7-diethyl-2,4,6,8-nonatetraenoic acid ethyl ester after chromatography on silica gel Hydrolysis as in Example 5 gave (all-E) - 9- [2-10 (8-hydroxyoctyl) oxy] phenyl-3,7-dimethyl-2,4,6, B-nonatetraenoic acid, m.p. 122-123 ° C (of ethyl acetate).

Example 12 (All-E) - [5- (2-nonyloxyphenyl) -3-methyl-2,4-pentadienyl] triphenylphosphonium bromide 62 g of 2- (nonyloxy) benzaldehyde dissolved in 500 ml of acetone was treated with 100 ml of 1M aqueous sodium hydroxide at room temperature for 18 hours.

Then brine and ethyl acetate / hexane (1: 1 parts by volume) were added. Concentration of the organic phase followed by crystallization of hexane afforded 53 g of 4- (2-nonyloxyphenyl) -3-buten-2-one.

A solution of 58 g of 4- (2-nonyloxyphenyl) -3-buten-2-one in 200 ml of tetrahydrofuran was added at -30 ° C to 200 ml of a 1.6M solution of vinyl magnesium bromide in tetrahydrofuran (diluted to 1 1 with more tetrahydrofuran). After completion of the addition, the mixture was stirred at 0 ° C for 30 minutes, quenched with 100 ml of saturated aqueous 25 ammonium chloride and 2 L of ether, and solids were filtered off. Concentration of the organic extract and purification by chromatography on silica gel gave 40 g of (E) -5- (2-nonyloxyphenyl) -3-hydroxy-3-methyl-1,4-pentadiene as an oil.

A solution of 66 g of (E) -5- (2-nonyloxyphenyl) -3-hydroxy-3-methyl-1,4-pentadiene in 250 ml of acetonitrile was added to a slurry of 66 g of triphenylphosphine hydrobromide in an additional 300 ml of acetonitrile at 10 ° C.

After warming to room temperature, the mixture was stirred at this temperature for 2 hours to give a solution. This solution was then extracted with 2 x 250 ml hexane and the acetonitrile phase was concentrated (about 400 ml) and cooled to -10 ° C. The solids were filtered off, washed with acetonitrile and hexane and dried to give 21 g of pure (all-E) - [5- (2-nonyloxyphenyl) -3-methyl-2,4-pentadienyl] triphenylphosphonium bromide .

EXAMPLE 13

Starting from (all-E) - [S- (2-nonyloxyphenyl) -3-methyl-2,4-pentadienyl] triphenylphosphonium bromide and using the procedure of Example 5, the ylid was reacted with 3-formyl-2 -butenoic acid ethyl ester to give (all-E) -9- (2-nonyloxyphenyl) -3,7-dimethyl-2,4,6,8-nona-tetraenoic acid after purification by chromatography on silica gel and hydrolysis with mock ethanolic potassium hydroxide solution as in Example 5.

EXAMPLE 14 (Ζ, Ε, Ε, Ε) -9- [2- (Nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid 10 g (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraacetic acid ethyl ester was dissolved in 200 ml of hexane containing 0.5 g of iodine and stirred at room temperature for 30 minutes. The hexane was washed free of 15 iodo with an aqueous sodium thiosulfate solution (10% by weight), dried and concentrated to give a mixture of double bond isomers. Separation by chromatography on silica gel gave 1.5 g of pure (Z, E, E, E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester. Hydrolysis with aqueous ethanolic potassium hydroxide under reflux gave pure (Z, E, E, E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, m.p. 136 ° C.

EXAMPLE 15 (Ε, Ε, Ε, Ζ) -9- [2- (Nonyloxy) phenyl] - 3,7-dimethyl-2,4,6,8-nonatetraenoic acid

The parent liquor resulting from the crystallization of 25 (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester in Example 5 was a mixture containing various isomers. Purification by chromatography gave an 80% pure ethyl ester which, after hydrolysis as in Example 5, gave pure (E, E, E, Z) -9- (2- (nonyloxy) p-henyl] -3,7-dimethyl-2, 4,6,8-nonatetraenoic acid, mp 105-109 ° C.

EXAMPLE 16 2-Decyl-1-bromobenzene 25.1 moles of nonylmethyltriphenylphosphonium bromide in 200 ml of tetrahydrofuran was converted to the ylide with 0.1 mole equivalent of n-butyllithium (1.6M in hexane) at -10 ° C.

0.09 mol of 2-bromobenzaldehyde was then added in 25 ml of tetrahydrofuran and after stirring for a further 30 minutes at 0 ° C, hexane and aqueous methanol were added (40:60). The hexane extract was concentrated and the residue was distilled to give 2- (1-decenyl) -1-bromobenzene (90%).

This material was dissolved in hexane containing a 10% palladium / -10 carbon catalyst and hydrogenated at room temperature and atmospheric pressure until the olefinic bond was saturated. The solids were filtered off and distillation of the residue gave pure 2-decyl-1-bromo-benzene (80%), boiling point 120 ° C (0.001 mm Hg).

EXAMPLE 17 2-Decyl-1-hydroxymethylbenzene 0.1 mole of 2-decyl-1-bromobenzene dissolved in 150 ml of ether was treated with 0.11 equivalents of n-butyl lithium (1.6M in hexane) and the mixture was stirred at room temperature. for 2 hours.

0.2 molar equivalents of dry paraformaldehyde was then added and the mixture was stirred for an additional 18 hours at room temperature.

Water and more ether were then added and the ether extracts dried and concentrated. After chromatography, the residue gave pure 2-decyl-i-hydroxymethylbenzene (75% yield).

EXAMPLE 18 (All-E) -9- (decylphenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid 2-Decyl-1-hydroxymethylbenzene was converted to the phosphonium salt with triphenylphosphonium hydrobromide in acetonitrile by the procedure of Example 1 This salt was then exposed to n-butyllithium in tetrahydrofuran as described above and then treated with 7-formyl-3-methyl-2,4,6-heptatrienoic acid methyl ester as described above.

26 DK 173287 B1

Purification of the crude condensation product by chromatography on silica gel followed by basic hydrolysis gave pure (all-E) -9- (decylphenyl) -3,7-dimethyl-2 (4,6,8-nonatetraenoic acid, m.p. 108 ° C (of hexane / ether).

EXAMPLE 19 (All-E) -9- (2-Octylaminophenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester 1 mole of 2-aminobenzyl alcohol was treated with 2.2 moles of octanoyl chloride in a mixture of dichloromethane and triethylamine at 0 ° C. After 30 minutes at 10 ° C, the mixture was washed with water and the ether was distilled off. The crude residue was dissolved in 2000 ml of tetrahydrofuran, treated with 1500 ml of 1 N aqueous sodium hydroxide and stirred at room temperature for 3 hours.

Addition of water and ether gave the crude hydroxymethyl octylamide. Purification by chromatography gave the pure octyl amide (85%).

100 g of this material was dissolved in 500 ml of tetrahydrofuran and added to a slurry of 2 molar equivalents of lithium aluminum hydride in 1000 ml of tetrahydrofuran. The mixture was then refluxed for 6 hours and cooled to 0 ° C and quenched with 100 mL of aqueous sodium sulfate solution.

The solids were filtered off, the solvents removed in vacuo and the residue purified by chromatography on silica gel to give 75 g of pure 2-hydroxymethyl-N-octylamaline.

This material was dissolved in 300 ml of acetonitrile containing 1.1 equivalents of triphenylphosphine hydrobromide and the mixture was refluxed for 24 hours and then concentrated. The residue was treated with ether to give the phosphonium salt as a white solid.

This material was converted to the corresponding ylide with 1.5 mol-30 equivalents of n-butyllithium and stirred at 0 ° C for 1 hour. 1.6 mole equivalents of excess 7-formyl-3-methyl-2,4,6-heptatrienoic acid ethyl ester were then added in tetrahydrofuran and the mixture was stirred at 10 ° C for 1 hour.

Adding hexane and aqueous methanol (2: 3) and removing hexane 35 in vacuo gave the crude coupled product. Purification by chromatography 271 on silica gel and crystallization of hexane gave pure (all-E) -9- (2- oc-methylaminophenyl) -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester (25%) , mp 38-40 ° C.

EXAMPLE 20 2- Fluoro-6-nonyloxybenzyl alcohol

A solution of 100 g of 3-fluorophenol in 1000 ml of dimethylformamide containing 165 g of potassium carbonate was treated with 115 g of allyl bromide and heated at 80 ° C for 18 hours.

Water and hexane were then added and the hexane extract was washed with 10 5% aqueous sodium hydroxide solution and saturated brine and evaporated to give 155 g of the allyl ether. This material (134 g) was heated at 220 ° C for 16 hours to give a mixture of 3-fluoro-2- (2-butenyl) phenol and 5-fluoro-2- (2-butenyl) phenol. This mixture was dissolved in 2000 ml of dimethyl formamide containing 170 g of 15 1-bromnonane and 150 g of potassium carbonate and heated at 80 ° C for 16 hours. Dilution with water and extraction with hexane gave a mixture of products after concentration. Distillation gave a mixture of 3 - [(2-fluoro-6-nonyloxy) phenyl] butene and 3 - ((3-fluoro-2-nonyloxy) phenyl] butene (186 g), bp 120-125 ° C (0.1 g) mm Hg).

This mixture of isomers (185 g) in 1000 ml of dimethyl sulfoxide containing 1.5 g of potassium tert-butoxide was left at room temperature for 6 hours. Addition of water and extraction with hexane gave a mixture of 1- [(2-fluoro-6-nonyloxy) phenyl] butene and 1- [(3-fluoro-2-nonyloxy) phenyl] butene.

This mixture of isomers (175 g) was dissolved in a mixture of dichloromethane and methanol (9: 1, 2000 ml) and exposed to an ozone stream at -40 ° C for 8 hours. Then, the reaction mixture was poured into a mixture of water, hexane and dimethyl sulfide (100 ml) and stirred at room temperature for 1 hour.

The hexane extract was washed with water, dried over MgSO 4, treated with an additional 50 ml of dimethyl sulfide and left at room temperature for 16 hours.

Removal of the solvents gave 155 g of a mixture of the aldehydes 2-fluoro-6-nonyloxybenzaldehyde and 4-fluoro-2-nonyloxybenzaldehyde.

28 DK 173287 B1

This mixture of aldehydes (150 g) in 2000 ml of ethanol was exposed to 15 g of sodium borohydride at 5 ° C and then stirred at room temperature for 30 minutes. 1500 ml of water and 500 ml of brine were added and the mixture of alcohols was extracted in hexane. Removal of the solvents and chromatography of the residue on silica gel (5% ethyl acetate / hexane mixture) gave 76 g of pure 2-fluoro-6-nonyloxybenzyl alcohol.

EXAMPLE 21 (All-E) - 9-2-fluoro-6 - (nonyloxy) phenyl-3,7-dimethyl-2,4,6,8-nonatetraenoic acid A mixture of 19 g of 2-fluoro-6- nonyloxybenzyl alcohol and 26 g of triphenylphosphine hydrobromide in 250 ml of acetonitrile were heated under reflux for 14 hours and then concentrated to dryness to give 42 g of [f (2-fluoro-6-nonyloxy) phenyl] methyl] triphenylphosphonium bromide. This phosphonium salt was dissolved in 600 ml of tetrahydrofuran, cooled to -50 ° C and treated with 45 ml of n-butyllithium (1.6M in hexane).

After a further 15 minutes of stirring at -50 ° C, 8.4 g of 7-formyl-3-methyl-2,4,6-octatrienoic acid ethyl ester was added and the reaction mixture was warmed to room temperature and stirred for a further 15 minutes. Hexane was then added and the mixture was washed with water and 40% aqueous methanol and dried over MgSCV. Concentration of the hexane extract and purification by chromatography (5% ether / hexane) gave 11 g of the pure trans isomer.

Crystallization of hexane / ethyl acetate gave 9.5 g (all-E) -9- [2-fluoro-6- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester.

A solution of 6.5 g of the ester in 150 ml of ethanol was treated with a solution of 7 g of potassium hydroxide in 40 ml of water and heated at reflux for 1 hour. The cooled reaction mixture was poured into cold aqueous hydrochloric acid and the acid extracted in chloroform. Removal of the solvents and crystallization of hexane / ethyl acetate gave pure (all-E) -9- (2-fluoro-6- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, m.p. 107-109 ° C.

EXAMPLE 22 Capsule Formulations

No. Ingredients mg / capsule tng / capsule mg / capsule 5 1 (all-E) -9- [2- 15 30 60 (nonyloxy) phenyl] - 3.7- dimethyl-2,4, - 6.8- nonatetraenoic acid 2 Lactose 239 224 194 10 3 Starch 30 30 30 4 Talc 15 15 15 5 Magnesium stearate 11 1

Capsule filling weight 300 mg 300 mg 300 mg 15 ---

Procedure: 1) 1-3 mix appropriately.

2) Talc and magnesium stearate are added and mixed for a short period of time.

20 3) Fill in capsules on an appropriate encapsulation machine.

EXAMPLE 23

Capsules are prepared by the method of Example 22 except that the active ingredient (# 1) was (all-E) -9-Γ2-2 fluoro-6- (nonyloxy) phenyl1-3,7-dimethyl-3 , 4,6,8-nonatetraenoic acid.

EXAMPLE · 24

Tablet formulation (wet granulation)

No. Ingredients mg / tablet mg / tablet mg / tablet 5 1 (all-E) -9-12- 250 250,500 (nonyloxy) phenyl] - 3.7-dimethyl-2,4, - 6.8- nonatetraenoic acid 2 Lactose 98.5 147, 5 170 10 3 Polyvinylpyrroli- 15 30 40 don (pvp) 4 Modified starch 15 30 40 5 Corn starch 15 30 40 6 Magnesium stearate 1.5 2.5 5 15 ----------------

Tablet weight 245 mg 490 mg 795 mg

Procedure: 1) 1, 2, 4 and 5 are mixed in an appropriate mixing machine, granulated with PVP and dissolved in water / alcohol. The granulation is dried. The dry granulation is ground through an appropriate mill.

2) Magnesium stearate is added and compressed on an appropriate pressing machine.

EXAMPLE · 25

Tablets are prepared in the same manner as in Example 24 except that the active ingredient (# 1) was (all-E) -9- [2-fluoro-6- (nonyloxy) phenyl] -3,7-dimethyl - 2,4,6,8-nonatetraenoic acid.

EXAMPLE 26 DK 173287 B1

Tablet formulations (direct compression)

No. Ingredients mg / tablet mg / tablet mg / tablet 5 1 (all-E) -9- [2- 15 eo (nonyloxy) phenyl] - 3.7-dimethyl-2,4, - 6.8- nonatetraenoic acid 2 Lactose 207 192 162 10 3 Avicel® 45 45 45 4 Direct Compression 30 30 30 Starch 5 Magnesium Stearate 3 3 3 15 Tablet Weight 300 mg 300 mg 300 mg

Procedure: 1) 1 is mixed with an equal amount of lactose. Mix thoroughly.

2) Mix with # 3, 4 and the remaining amount of # 2.

Mix thoroughly.

3) Magnesium stearate is added and mixed for 3 minutes.

4) Compress on an appropriate piston.

EXAMPLE 27 25 Capsule Formulations

No. Ingredients mg / capsule mg / capsule 9 / capsule 1 (all-E) - (3,7-di-60-methyl) -9- [2 - [(8-30 hydroxyoctyl) oxy] -phenyl] 2,4,6,8-nonatetraenoic acid 2 Lactose 239 224 194 3 Starch 30 30 30 35 4 Talc 15 15 15 5 Magnesium stearate li 1

Capsule filling weight 300 mg 300 mg 300 mg 32 DK 173287 B1

Procedure: 1) 1-3 are mixed in an appropriate mixing machine.

2) Talc and magnesium stearate are added and mixed for a short time.

5 3) Fill in capsules on an appropriate encapsulation machine.

EXAMPLE 28

Tablet formulations (wet granulation)

No. Ingredients mg / tablet mg / tablet mg / tablet 10 -----—- 1 (all-E) - (3,7-di-100,250,500 methyl) -9- [2 - [(octyl-oxy) methyl ] phenyl] - 2,4,6,8-nonatetraene-acid 2 Lactose 98.5 147.5 170 3 Polyvinylpyrrolidone 15 30 40 4 Modified starch 15 30 40 5 Corn starch 15 30 40 20 6 Magnesium stearate 1.5 2.5 5

Tablet weight 245 mg 490 mg 795 mg

Procedure: 1) No. 1, 2, 4 and 5 are mixed in an appropriate mixing machine, granulated with PVP and dissolved in water / alcohol. The granulation is dried. The dry granulation is ground through an appropriate mill.

2) Magnesium stearate is added and compressed on a suitable press machine.

EXAMPLE 29 t [2-Trifluoromethyl-S - (nonyloxy) phenyl] methyl] triphenylphosphonium bromide

A mixture of 51 g of α, or α, trifluoro-c-cresol, 70 g of 1-bromnonane and 100 g of potassium carbonate in dimethylformamide was heated at 85 ° C for 48 hours. Addition of water and hexane gave 89 g of pure (3-trifluoromethylphenylnonylphher, boiling point 115 ° C (0.1 mm Hg). 89 g of this product in 1.5 L ether at -20 ° C was mixed with 233 ml of n -butyl lithium (1.5M in hexane) and then stirred for 2 hours at room temperature.This mixture was then cooled to -40 ° C, treated with excess dry dimethylformamide (40 ml) in 100 ml of ether, heated to 0 ° C and then treated with water Extraction with hexane and chromatography on silica gel (5% ether / hexane) afforded 35 g (2-trifluoromethyl-6-nonyloxy) benzaldehyde Reduction of this product 10 with sodium borohydride in ethanol at The procedure described in Example 1 gave 32 g of (2-trifluoromethyl-6-nonyloxy) benzene methanol after chromatography on silica gel 31 g of this material was converted to [[2-trifluoromethyl-6- (nonyloxy) phenyl] methyl] triphenyl phosphonium bromide by reaction. with triphenylphosphine hydrobromide by the procedure described in Example 1.

EXAMPLE 30 (All-E) -9- [2- (trifluoromethyl) -6- (nonyloxy) phenyl] -3,7-dimethyl-2.4.6.8-nonatetraenoic acid 97 mmol [[2-trifluoromethyl-6- (nonyloxy) phenyl ] methyl] triphenylphosphonium bromide in 600 ml of tetrahydrofuran was reacted with (all-E) -9- (2- (trifluoromethyl) -6- (7-formyl-3-methyl-2,4,6-octatrienoic acid ethyl ester) nonyloxy) phenyl] -3,7-dimethyl-2.4.6.8-nonatetraenoic acid ethyl ester by the procedure described in Example 3. Purification by chromatography and crystallization of hexane gave the pure ethyl ester (41%). Hydrolysis (5.2 g) as in Example 5 gave 3 g of pure (all-E) -9- [2- (trifluoromethyl) -6- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, mp 135-136 ° C (of ethyl acetate / hexane).

Example 31 (All-E) -9- [2- (Hexyloxy) phenyl] - 3,7-dimethyl-2,4,6,8-nonatetraene acid [[2- (Hexyloxy) phenyl] methyl] triphenylphosphonium bromide was prepared by the procedure described in Example 1 by reaction of 2-hydroxybenzaldehyde and 1-bromohexane, was converted to (all-E) -9- (2- (hexyloxy) phenyl] -3,7-dimethyl-2,4 , 6,8-nonatetraenoic acid, mp 137-138 ° C (of ethanol) by the procedure described in Example 3.

EXAMPLE 32 34 DK 173287 B1 [[2- (Nonyloxy) -S- (hydroxy) phenyl] methyl] triphenylphosphonium bromide

A solution of 1 mole of 4-bromophenol in 500 ml of tetrahydrofuran was added to a slurry of 1.17 moles of sodium hydride in 1.2 l of dimethylformamide at 25 ° C. After completion of the reaction, 1.32 moles of allyl chloride were added and after a further 3 hours stirring at 45 ° C the product was isolated with water and hexane. Distillation gave allyl- (4-bromophenyl) ether, bp 65-67 ° C at 0.1 mm Hg (82%). This material was heated at 195 ° C with dimethylanaline for 4 hours and then distilled to give 0.81 mol of 2-allyl-4-bromophenol.

A solution of 0.81 mole of this material in 200 ml of tetrahydrofuran was added to a mixture of 0.8 mole of 1-bromnonane, 0.92 mole of sodium hydride and 1 g of potassium iodide in dimethylformamide at 25 ° C.

After the hydrogen evolution was complete, the mixture was heated at 50 ° C for 14 hours, cooled, added to an excess of water and extracted with hexane. Distillation gave 256 g of nonyl (2-allyl-4-bromophenyl) ether, bp 147-156 ° C at 0.1 mm Hg. 255 g of this material in dimethylsulfoxide and 0.5 l of tetrahydrofuran were heated at 35-40 ° C with 2 g of potassium tert.butoxide for 2 hours, after which the reaction was quenched with 5 ml of acetic acid and water.

Isolation of the reaction products with hexane gave 234 g of pure 1- [2- (nonyloxy) -5- (bromo) phenyl] propylene, bp 145-155 ° C at 0.1 mm Hg. A solution of 0.56 mol of this material in 600 ml of tetrahydrofuran was converted to the Grignard reagent with 1 mol of magnesium 25 at 55 ° C for 3 hours. After completion of the reaction, the mixture was cooled to 0 ° C and treated with 0.75 mole of trimethylborate in 200 ml of ether.

After a further 30 minutes of stirring at 25 ° C, the mixture was cooled to 0 ° C and exposed to 500 ml of a mixture of 10% ammonium chloride and 10% hydrogen peroxide and stirred for an additional 30 hours at 25 ° C. Addition of water and hexane gave the crude material after removal of the hexane in vacuo. The crude product was passed through a plug of silica gel to give 73 g of para- [2- (1-propenyl-4- (nonyloxy) phenyl] phenol. Acetylation of 0.8 g of this material with acetyl chloride and triethylamine in dichloromethane gave [2 - (1-35 propenyl) -4- (nonyloxy) -1- (acetoxy)] benzene (89%) 99 g of this material was dissolved in a mixture of 150 ml of methanol and 1.5 l of dichloromethane and treated with ozone at 40 ° C until all starting material was used, 50 ml of dimethyl sulfide and 500 ml of water were then added and after vigorously stirring for 30 minutes at 40 ° C, the organic phase was dried over MgSO 4 and concentrated to give 83 g 2- (nonyloxy) -5- (acetoxy) Ibenzaldehyde Reduction of 80 g of this material with 6 g of sodium borohydride in ethanol at 20 ° C for 2 hours gave crude [2- (nonyloxy) -5- (acetoxy) 1benzene methanol. DK 173287 B1 nol, which was immediately exposed to 300 ml of 40% aqueous potassium hydroxide in ethanol for 30 minutes at 60 ° C. Acidification with aqueous acid (6M hydrogen chloride) and extraction with ch. loroform gave the crude product after concentration. Dilution of the residue with hexane gave 5 63 g of pure [3- (hydroxymethyl) -4- (nonyloxy)] phenol as a solid.

A solution of 62 g of this material in a mixture of 0.5 l of acetonitrile and 86 g of triphenylphosphine hydrobromide was heated under reflux for 14 hours and concentrated to dryness at 50 ° C to give in [2- (nonyloxy) -5- (hydroxy) phenyl] methyl] triphenylphosphonium bromide in the form of a glass.

EXAMPLE 33 (All-E) -9- [5-hydroxy-2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid 0.23 mol Γ [2- (nonyloxy) - 5- (hydroxy) phenyl] methyl] triphenylphosphonium bromide in 1.5 L of tetrahydrofuran at -70 ° C was treated with 315 ml of n-butyllithium (1.6M in hexane) and then treated with 59 g of ethyl 8-formyl -3,7-dimethyl-2,4,6-octatrienoate in tetrahydrofuran. The mixture was then heated to -15 ° C, acidified with acetic acid and extracted into ether and 40% aqueous methanol. Purification by silica gel chromatography gave pure (all-E) -9- [5- hydroxy-2- (non-nyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester . Hydrolysis of 6 g of this ester by the procedure described in Example 5 gave 3.5 g (all-E) -9- [5-hydroxy-2- (nonyloxy) phenyl] -3,7-dimethyl-2,4, 6, B-nonatetraenoic acid, m.p. 170-173 ° C (of ethyl acetate).

Example 34 (All-E) -9- [2- (nonyloxy) -5- (2,2,2-trifluoroethoxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid 4.4 g (all-E) -9- [5-hydroxy-2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester was heated at 90 ° C for 72 hours with 7 g of potassium carbonate and 6 g of 2,2,2-trifluoroethyl-p-toluene sulfonate in 200 ml of dimethyl formamide. Work-up with water and hexane followed by purification on silica gel gave 0.75 g of the pure ethyl ester. Hydrolysis of 0.9 g of this ester by the procedure described in Example 5 gave 0.6 g of pure (all-E) -9- [2- (nonyloxy) -5- (2,2,2-trifluoroethoxy) phenyl ] -3,7-Dimethyl-2,4,6,8-nonatetraene acid, melting point 121 ° C, after crystallization of a mixture of tetrahydrofuran and hexane.

EXAMPLE 35 (2) -t [2- (1-Decenyl) phenyl] methyl] triphenylphosphonium bromide and 5 (E) - {[2- (1-Decenyl) phenyl] methyl] triphenylphosphonium bromide

An (E, Z) mixture of 2- (1-decenyl) -1-bromobenzene prepared according to Example 16 (1: 4) was converted to an (E, Z) mixture of 2- (1-decenyl) -1-hydroxymethylbenzene by the process described in Example 17. This mixture was separated by chromatography on 10 silica gel to give the pure (E) and (Z) alcohols. Reaction of each of these isomers with triphenylphosphine hydrobromide as described in Example 1 gave the corresponding phosphonium salts, i.

(2) - [(2- (1-decenyl) phenyl] methyl] triphenylphosphonium bromide and (E) - [[2- (1-decenyl) phenyl] methyl] triphenylphosphonium bromide.

Example 36 (All-E) -9- [2- (1-decenyl) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraene acid (E) - [(2- (1- The decenyl) phenyl] methyl] triphenylphosphonium bromide was converted to the ethyl ester of (all-E) -9- [2- (decenyl) phenyl] -3,7-20 dimethyl-2,4,6,8-nonatetraenoic acid in the example of Example 1. base hydrolysis as in Example 1 and crystallization of the crude acid of acetonitrile gave (all-E) -9- [2- (1-decenyl) phenyl] -3,7-dimethyl-2,4,6 , 8-nonatetraenoic acid, mp 105-107 ° C.

Example 37 (E, E, E, E, Z) -9- [2- (1-Decenyl) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid

The title compound was prepared in the same manner as in Example 36 using (Z) - [[2- (1-decenyl) phenyl] methyl] triphenylphosphonium bromide. Hydrolysis of the ethyl ester and crystallization of the crude acid of ether gave pure (E, Ε, Ε, E, Z) -9- [2- (1-decenyl) phenyl] -3,7-dimethyl-2.4, 6,8-nonatetraenoic acid, mp 103-105 ° C.

In the examples below, compound A (all-E) -9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6, B-nonatetraenoic acid. In the examples below, compound A was tested in various experiments for its anti-inflammatory activity in animal models of inflammation and in 5 chronic models of adjuvant arthritis.

In all the experiments, compound A and the other retinoids tested simultaneously were formulated in peanut oil containing 0.05% propyl gallate as antioxidant. Dosage rates used were 5 ml.kg 'for rats and 10 ml.kg for mice. Controls were dosed with the appropriate amount of peanut oil carrier.

EXAMPLE 38

Effect of Compound A on Delayed Hypersensitivity to Methylated Bovine Serum Albumin (MBSA)

Animals·. MFI male and female mice, sub strain E33. Starting weight approx.

25 g.

Materials: Methylated bovine serum albumin (MBSA) (Sigma).

Freund's complete adjuvant (Difco).

Method: Groups of 10 mice were sensitized (day 0) by intradermal injection at two abdominal sites of 0.05 ml of 20 of a water-in-oil emulsion of MBSA and Freund's complete adjuvant. On day 9, mice were infected by injecting 20 μΐ of a 1% MBSA solution into one paw and 20 μΐ water into the contralateral paw. Paw volumes were measured 24 hours later by mercury displacement plethysmography. The mean percentage increase in the paw volume of the MBSA-infected paw compared to the water-infected paw was calculated for each treatment group. Carrier and retinoid dosing began on day 0 and 30 ended on day 9.

Results: The results are shown in Table II below.

38 DK 173287 B1

Table II

Effect of Compound A in MBSA Delayed Hypersensitivity Test 5 Treatment Dose% Increase in% Reduction Average mg. kg'1 paw volume (cf. essential oil control) reduction of body weight (g) 10 -: -—

Peanut oil 109 ± 11 M 3.8 F 0.2

Etretinate 10 59 ± 9 ** 46 M -0.8 F -2.0

TiS

Compound A 10 102 ± 12 6 M 3.3 F -0.2

Compound A30 50 ± 7 + ** 54 M2.8 F 0.5

Compound A 100 40 ± 5 *** 63 M 3.3 20 F -0.2

Each group consisted of 4 male noses and 6 female mice (in separate cages). The drugs were administered orally in a dose amount of 10 ml.kg "1 (10 doses).

25 ns: Not significant **: p <0.01 ***: p <0.001 compared to carrier control using "Student's two-tailed t test".

Example 39 The effect of compound A on the development of adjuvant arthritis in rats

Animals: AHH / R female rats (PVG-derived with an initial weight in the range of 110-140 g) were used.

Materials: Adjuvant for injection. A homogenized suspension of heat-killed Mj tuberculosis (human strains C, DT and PN), 5 mg.ml'1 in liquid paraffin was prepared.

Method: The rats were randomly divided into groups of five and adjuvant arthritis was induced by subplantar injection of 0.1 ml of adjuvant suspension in the right hind paw of each rat. The test compounds were administered by intubation each morning starting with the day of adjuvant injection. Two groups of control rats received carrier 39, as did a group of three normal rats that were included for comparison purposes. Dosage was taken daily until the end of the trial on day 15 except for the first weekend (days 5 and 5 6). The treatment groups are shown in Table III and have etretinate as standard retinoid.

Measurements of right hind paw volume were made initially and on days 2 and 4 after adjuvant injection (primary phase). The right and left hind paw volumes were then measured on day 8 and every two or three days until the end of the trial on day 15 (secondary phase). At this time, the mobility of each ankle joint and the incidence and severity of secondary lesions on muzzle, ears, forepaws, left hindpaw and tail 15 were also assessed for the degree of possible flexion and using an arbitrary scoring system.

Evaluation of results:

Time course curves for the injected paws were integrated from days 0 to 4 to reflect primary swelling and from days 8 to 15 (secondary swelling). The secondary swelling of the uninjected paw was similarly integrated from days 8 to 15. Calculations were performed using a specific computer program that calculated averages ± see for each integrated area. The significance of the differences relative to controls was determined by Student's t-test (2-tailed) and percentage reductions in control ranges were calculated. Percentage improvements in joint mobility and percentage reductions in lesion degree were also determined. In the latter case, "Wilcoxon rank sum test (2 tailed)" was used to express the difference to the degree of control using raw data. The mean change in body weight in each group was recorded.

Results: The results are listed in Table III below.

40 DK 173287 B1

Table III

Efficacy of Compound A and Eretin Retinate P & Adjuvant Arthritis High Rats

Treatment Dose \ reduction of paw swelling * Lesions- ^ Arthritis- ® Cndration 1 mg.kg * Primary Secondary Secondary degree Degree Oxidized body heat (j) Oral (right) (left) (1NC) ----- y -—-

You U-i!> Day 0-15 Check "♦» # 3 e 3.7

Adjuvant - - - - - - - 0.6-5.5 control *

Compound A 15 -6 30 91 · 72 66 * 2i2 - 0, fc 65 '16 5l "' 75" 65 * 66 '* 5, *, * 1,2'

Crtrtlla '! 15 9. 30 67 * 56 * 50 ♦ 1.0 - 5.0 1) Statistical analysis by Student's t-test (2-tailed) 5 2) Statistical analysis by Wilcoxon rank sum test (2-tailed) * = P <0.05 , ** = p <0.02, *** »p <0.01 EXAMPLE 40

The effect of compound A on established type II collagen arthritis

Animals: Alderley Park Strain 1 male and female rats.

10 Materials · Type 2 collagen (made from bovine nasal septum cartilage), Freund's incomplete adjuvant (Difco).

Method: The rats were sensitized to type 2 collagen by intradermal injection with 1 ml of a water-in-oil emulsion consisting of equal parts of a 1 mg.ml / 15 solution of type 2 collagen in 0.45M NaCl, 0.02M Tris, pH 7.4, and Freund's incomplete adjuvant. Rats developing arthritis were allocated on day 15 after sensitization to an arachis oil treated control group (6 male rats, 4 female rats) or to the 20 group treated with compound A (6 male rats, 5 female rats). Rear foot volume measurements were performed to ensure equal distribution of rats between groups. Urine collection was performed overnight between days 15 and 16 and dosing commenced on day 25 16. These urine samples were analyzed for glycosaminoglycans (GAG). Compound A was given with 100 mg-kg'1 orally. On the night between days 19 and 20, another collection of urine was made. These urine samples were analyzed for glycosamine glycans (GAG). On day DK 173287 Bl 41 20, another measurement of the hind paw was made.

The rats were then anesthetized with sodium pentobarbitone, annualized and sacrificed, and radiographs of the hind and forepaws were taken. The rats were dosed on 5 days 16-19 incl. (4 doses).

Results: The results are shown in Table IV below.

Table IV

The Effects of Compound A on Type II Collagen Arthritis Test 10 ---

Day Compound A Control 100 mg. kg "1 Peanut oil

Potevol. GAG ^ g) weight (g) Potevol. GAG ^ g) Weight (g) 15 16 2.44 + 0.08 15B6 ± 307 247 * 14 2.48 * 0.07 1538 * 192 249 * 19 20 2.66 ± 0.06 634 * 72 229 * 10 2.46 ± 0.07 729 ± 134 258 ± 19

Comment: Both male and female rats lost weight during treatment with compound A.

EXAMPLE 41

Effect of Compound A on Non-Immune Inflammation Animals ·. Alderley Park Strain 1 female rats weighing 170-205 g were used at the beginning of the experiment.

Materials: λ-carrageenan. Prepared as a solution in saline 25 and sterilized by autoclaving.

Method: Compound A was administered orally to groups of 8 rats once daily for 10 days at doses of 10, 30 and 100 mg.kg'1. Control animals received carrier. One hour after the last dose, the animals were anesthetized with methohexitone 30 (Brietal, 50 mg.kg'1) and 0.2 ml of 1% 1-carrageenan was injected into the pleural cavity. Four hours later, the animals were sacrificed with an overdose of pentobarbitone (Sagatal), the pleural exudate was collected, and the pleural cavity was washed out with 2 ml of phosphate-buffered saline (PBS-A, Oxoid). The exudate volume was recorded and cell counts were determined with an automatic cell counter (Coulter). Differential cell counts were made on exudate streaks stained with Giemsa stain to separately determine the number of polymorphonuclear leukocytes (PMN) and mononuclear cells. (MN).

Immediately after collecting the pleural exudates, the tibia of the animals was excised and their rupture strength was determined.

The body weight of the animals was recorded daily. Statistical analyzes were performed using Student's 2-tailed t-test.

Results: The results are given in Table V. The dose is given in mg per mg. kg per day.

Table V

The effects of compound A on the development of 4 hours carrageenan mesothelioma -. SMl.t Skliwi * b * n · - rorblixwi «Oo * I» YOlUMR e * ll «t» l 5 «i« Ρ * β * Sul * C tw bride »tyrk * (pcrorelt) fk * jr *» v * ni? r «) (al) t« 10 * «« 10 * t «10 * IC * i> r« i. «nlt» mdrlng andrlno »utring atidrlno (» 9! undr 1 * 9

Control 1 * 1 1.40 139.1 119.1 10.1 C, 7 liridilwll ·) «0.1 * 0.1 O,» * 0.) * 0.1 rorblnd.1 «. λ «·» ·. »« I10, i "s -4, S ΠΙ, *" * · -5, «0.4H k · -17.4», SH * · -1.4 (solution) * 0 , 00 «7.3« *, 7 * 0.0 0.1 rorbUd.1 .. λ o, »·· -> *,» in.i «-11,1» · «, * · -11, <S · "· * · '1.1« 0.09 0.0 O · 0.7 * 0.1 7o7bUd.l. «K>“ * »0.00 ·· -41.9 109, l" · 1 · -19.7 ΙΟΙ, * - * · -14.7 Ί ~ Γ · -7 ', 7 *, 7 "* · 0.1, 0.00 * 0.1« 7.1 «1.1 «0.1 NS: Not significant 15 * ap <0.05, ** = p <0.01 compared to carrier-treated controls using Student's 2-tailed t test.

EXAMPLE 42

Effect of Compound A on Impregnated Mushroom Granuloma Test in Rats 20 Animals: AHH / R female rats (PVG derived) with a starting weight in the range of 120-140 g were used.

Materials: Mushroom preparation. Pellets (6.5 mm in diameter) were punched from cellulose sponge cloth ("Wettex") and 0.1 ml of a suspension containing 0.5 mg.ml'1 heat-killed M 2. tuberculosis (human strains C, DT and PN) in sterile saline was applied to each pellet. Pellets were dried, weighed and autoclaved.

43 DK 173287 B1

Method: The rats were randomly divided into groups of five and daily dosing with test compounds started.

After the fifth dose, the rats were anesthetized with Saga number (45 mg.kg'1 intraperitoneally). The spines were shaved and two pellets were implanted subcutaneously (one on each side) in each rat via a small dorsal midline incision. The incision was closed and the rats were allowed to wake up from the anesthetic.

7 days after implantation, the rats were sacrificed and 10 pellets were removed, dissected free of foreign tissue and weighed. Each pellet was then placed in a 4 ml aliquot of distilled water, cut into fine scissors and sonicated. After centrifugation, the Na + and K + contents of the supernatant were determined by flash photometry. In addition, the adrenal gland and thymus gland from each rat were dissected and weighed, and the lower back bones were removed to measure the tibial fracture strength.

Body weights were also recorded throughout the trial period.

Results: The results are listed in Table VI below.

Assessment of the results

The mean ± SE for each of the parameters was calculated and differences relative to the control values were determined by Student's t-test (2-tailed). Percentage reductions in granuloma weight, Na + and 25 K + content and percentage changes in adrenal and thymus, weight and tibial fracture strength were determined.

Table VI

The effect of compound A, etretinate and dexamethasone on the impregnated fungal granuloma test in rats «action 0» · * · »reduction of (..i.ulo. rartar btnyro- thyeui »- aklnnabana- body * vo9t [91

VidVTdt Must »Κ» v «* t» edt bride-dry control · ------. », 4.1.1 rudder tie · * AI * por- ^ s« Μ "» «,» ·· - *, * ·· .7,1.0,74 45 Ρ · Γ * * l0 »· no, · .1», 0 *. »14.1 -1.1» .11,4.0, »« orally 9 * * 1 - 1

Circulation night IS W 4 1.7 5, * O, »07.» ·· * 11.7 -4.4 01,) 0.4 orally 1 '' ”ηι · ΜΜ (). Ι · οη 0.5 mb- s 4S # S · * · S! # I ·· * 75.7 »·· -44.S ··· -TV1 · -11.5 * cutaat r 9 · · * 1 44 DK 173287 B1 * p = <O, OS, ** p = <0.02, *** p = <0.01

Comment: 2 control rats and etretinate-treated rat died during the anesthesia.

Animals: Lewis River male rats were used for 5 of these experiments.

Materials: Heat-killed, dried Mycobacterium butvricum.

Method: Adjuvant arthritis was induced by injection of 0.1 ml of adjuvant (a suspension of heat-killed, dried Mvcobac terium butvricum. 0.5% (w / v) in heavy mineral oil containing 0.2% digitonin) at the halo. Arthritis was allowed to develop for 21 days, after which the volume of both hind legs was measured with a mercury plethystnograph. The rats were divided into groups of 8 with equal mean paw volumes, and then the rats were treated with compound A, indomethacin (as a control drug) or carrier for 7 days. At the end of the treatment period, the volume of both hind legs was again measured to assess the anti-inflammatory effects. Changes in body weight were also followed, and at the end of the experiment, plasma was collected to determine plasma fibrinogen (Exner et al., Amer. J. Clin. Path. 71. pp. 521-527).

Results: The results are given in Table VII below.

Table VII

a 25 Effect on the course of established adjuvant arthritis Am. in Plasma- Am. in

Treat- Dose potable. fibrinogenkropsvægt

Group lingb (mg / kg) (ml) e (mg / dl) (g) C

30 _____

Arthritis (10) d Carrier + 0.53 + 0.08 1773 ± 30 7.5 ± 1.2

Arthritis (10) Forb. A 100 -0.96 ± 0.10 * 874 ± 57 * 5.2 ± 2.1 Arthritis Indome- (10) Thacin 1 -1.22 + 0.15 * 978 ± 100 * 25.6 ± 3, 1 * a) Average ± SE is listed.

b) The drugs were administered once daily for 7 days at 40 intubation beginning on day 22 after induction of scar thritis. Tween® 80 was used as a carrier.

c) Change in paw volume / body weight is equal to paw volume / body weight on day 28 minus paw volume / body weight on day 22.

Claims (19)

1. Phenylnonatetraenoic acid derivatives of the general formula I Ϊ1 * 7 * · R CH «CH-C * CH-CH = CH-C» C H-Rs 0 | In x-r4 where R 1 represents hydrogen, chlorine, fluorine or trifluoromethyl; R 2 represents hydroxy, lower alkoxy, trifluoromethyl-lower alkoxy or hydrogen; R 4 represents alkyl of equal chain length pA 4 to 9 carbon atoms or -Cl 2 Cl 2 IdCH 2 OH; x represents -CHO-. -CH-. -ISLAND-. -C-CH- or -N-: R10 R10 R10 R10 R5 represents COOR9; 15 and R7, Rg, R9 and R10 represent hydrogen or lower alkyl; n is 6 or 7; and pharmaceutically acceptable salts thereof, wherein R 9 represents hydrogen. A compound according to claim 1, characterized in that R (represents hydrogen, chlorine or fluorine; R 1 represents hydrogen or lower alkoxy, R 4 represents alkyl of 8-9 carbon atoms having a straight chain length of 8 or 9 carbon atoms; -CH-0 X represents IL R10 Rio 10 Rj represents COOR9, and 25 n, R7, Rg, R <j and R have the meaning set forth in claim 1, and salts thereof, where hydrogen represents DK 173287 B1 A compound according to claim 1 or 2, characterized in that R 4 represents alkyl as defined in these claims. A compound according to claim 35, characterized in that X represents -O-. A compound according to claim 4, characterized in that R 1 represents chlorine or fluorine. A compound according to claim 4, characterized in that R 2 represents lower alkoxy. A compound according to claim 1, characterized in that the compound is (9- [2-chloro-6- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid. A compound according to claim 1, characterized in that the compound is (all-E) -9- [2-fluoro-6- (nonyloxy] phenyl] -3-7-dimethyl-2,4,6,8-nonatetraenoic acid . A compound according to claim 1, characterized in that the compound is 3,7-dimethyl-9- (5-methoxy-2-nonyloxyphenyl) -2,4,6,8-nonatetraenoic acid. A compound according to claim 1, 20, characterized in that the compound is 9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid. A compound according to claim 1, characterized in that the compound is (all-E) -3,7-dimethyl-9- (2-octylaminophenyl) -2,4,6,8-nonatetraenoic acid. Compound according to claim 1, characterized in that the compound is (all-E) -3,7-dimethyl-9- [2 - [(8-hydroxyoctyl) oxy] phenyl] -2,4,6,8 -nonatetraensyre. DK 173287 Bl A compound according to claim 1, characterized in that it is selected from (all-E) -8- [2- [2- (trifluoromethyl) -6- (nonyloxy) phenyl] -3,7-dimethyl-2, 4,6,8-nona-tetraenoic acid, 3,7-dimethyl-9- [2- (octyloxy) phenyl] -2,4,6,8-nonatetraenoic acid, 3.7-dimethyl-9- (2- (2, 2-dimethyloctyl) oxy] phenyl] -2,4,6,8-nonatetraenoic acid, 9- [2- (nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid ethyl ester, 10 ( all-E) -9- [2- (hexoxyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, (all-E) -9- [5-hydroxy- (2-nonyloxy) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid, (all-E) -9- [2- (nonyloxy) -5- (2,2,2-trifluoroethoxy) phenyl] -3, 7-15 dimethyl-2,4,6,8-nonatetraenoic acid, 3.7-dimethyl-9- [2 - [(octyloxy) methyl] phenyl] -2,4,6,8-nonatetraenoic acid, 9- (decylphenyl) -3 , 7-dimethyl-2,4,6,8-nonatetraenoic acid, 3.7-dimethyl-9- (2-octylaminophenyl) -2,4,6,8-nonatetraenoic acid ethyl ester and (all-E) -9- [2 - (1-decenyl) phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid. Compounds according to claim 1 for use as antirheumatic, antiarthritis and immunosuppressive agents. Compounds according to claim 14, characterized in that it is 9- [2- (nonyloxy) phenyl] -3,7-diraethyl-2,4,6,8-nonatetraenoic acid. Process for the preparation of compounds of the general formula in accordance with claim 1 or pharmaceutically acceptable salts thereof, characterized in that a) a compound of the general formula IIR, iOC in XR,. ψ is reacted with a compound of the general formula VII or 17) a compound of the general formula XVI R, R7 Y Ro I * I7 | Z \ A ^ CH = CH-C = CH-CH2 P-VZ 'OR v 5 is reacted with a compound of general formula XVII R' 0 XVII i8 'OHC-C ^ CH-C-OR1g or c) a compound having the general formula VIII R, R7 Rq 0 ri i7 i8 i "2 I, CH = CH-CH = CH-CH = CH-C = CH-C-OR'Q OH 10 is reacted with a compound of the general formula R4Z, wherein R 1, R 2f R 4, R 7 and R 9 in the above formulas bore the designation of claim 1; 1 * 9 'represents lower alkyl; Y represents aryl; Z represents a leaving group; and Z' represents a halide ion; whereupon, if desired, a carboxylic alkyl ester group COOR9 contained in the reaction product is converted to the free acid and, further, if desired, converted the acid to a pharmaceutically acceptable salt. Pharmaceutical compositions, characterized in that they contain a compound according to claim 1 and a carrier. Use of a compound according to claim 1 for the preparation of a pharmaceutical composition for use as an antirheumatic, anti-arthritis or immunosuppressive agent. Use according to claim 18, characterized in that the compound is 9- [2- (nonyloxy) -phenyl] -3,7-dimethyl-2,4,6,8-nonatetraenoic acid.