PT95305A - PROCESS FOR THE PREPARATION OF MACROCYCLIC COMPOUNDS - Google Patents

Description

Description of the patent of invention of PI SONS PLC, British, industrial and commercial, with headquarters in Pison House, Princes Street, Ipswich, Suffolk Ipl 1QH, England, (inventors: David Keith Donald, Mark Furber, David Norman Hardern and Paul Leff, residing in England), to " PROCESS FOR THE PREPARATION OF MACROCYCLIC COMPOUNDS11

The present invention relates to novel pharmaceutical uses of certain known macromeric compounds and to novel macrocyclic compounds having the same utility. European Patent Application No. 184162 (to Fujisawa Pharmaceuticals Co Ltd) discloses various macrocyclic compounds which are derivatives of 12- (2-cyclohexyl-1-methylvinyl) -13,19,21,29-tetramethyl- 28-dioxa-4-azatricyclo [22.3.1.0] octacos-18-ene (numbers PR-90056, PR-900520 and FR-900525) and isolated from microorganisms belonging to the genus Streptomvces . Macrocyclic compounds and some of their derivatives are indicated as immunosuppressive agents. - 1 -

International Patent Application No. WO 89/05304 (as well as European Patent Application No. 323042 to Pisons plc) discloses a large number of macrocyclic compounds which may be derived from those disclosed in European Patent Application No. 184162. They indicate The compounds are, first, as immunosuppressive agents.

European patent applications Nos 349049 and 349061 (both to Merck & Co Inc) each disclose a macrocyclic compound which is indicated as an immunosuppressive agent.

The compounds and methods disclosed in the above-mentioned patent applications in the production of the novel compounds of the present invention may alternatively be produced, the compounds may be produced by total synthesis.

A new group of macrocyclic compounds which act as antagonists of immunosuppressive compounds, especially immunosuppressive macrocyclic compounds including 12- (2-cyclohexyl-1-methylvinyl) -13,19,21,2,2-tetramethyl derivatives have now been discovered, -1,2,8-dioxa-4-9-azatricyclo [2.2.3.1.0] ocfcaeos-18-ene and rapamycin, as is seen in the mixed lymphocyte reaction (MLR) (described in WO 89/0304, Example A). The new group of compounds is, however, useful for example in the treatment of immunodepression or of a disorder involving immunodepression.

Thus, according to the present invention there is provided the use of a compound of formula I,

J

HO

wherein R1 and R2 independently represent H or OH, or may together represent a second carbon-carbon bridge between the carbon atoms to which they are attached. R3 represents methyl optionally substituted by -C02H or an ester thereof or an amide thereof; ethyl optionally substituted by 0, OH or -OH or an ester or amide thereof; propyl optionally substituted by OH or O; or ally optionally substituted by OH;

R4 is H and R5 together represent a second carbon-carbon bridge between the carbon atoms to which they are attached; R7 is H or OH; R6 is OCH8; R8 is OH or o3 is X or (η, ηη)? Y represents 0 or (H? OH); n is 1 or 2; And in addition to their aforementioned meanings R and R may together represent an oxygen atom. In this case R 6 and R 5 together represent a second carbon bond bridge between the carbon atoms to which they are attached; R 7 and R 6 may together represent an oxygen atom; and 34, R, R and Y, together with the carbon atoms to which they are attached, may represent a substituted furanyl-methyl ring; provided that in the manufacture of a medicament for the treatment of immunodepression or of a disorder involving immunodepression: i) when R represents H; R represents methyl,

ethyl, propyl or allyl; R and R represent a second carbon-carbon bridge between the

to which they are linked? R

represents OCHO? and Y is 0? thus, R being OH; and

Ii) when n is H1, then R3 is not methyl or ethyl.

Examples of disorders involving immuno-depression include AS | DS (AIDS), carcinoma, senile dementia, trauma (including wound dressings, surgery and shock), chronic bacterial infections and certain disorders of the central nervous system. The immunosuppressive to be treated may be caused by an excessive dose of an immunosuppressive macrocyclic compound, for example the 12- (2-cyclohexyl-1-methylvinyl) -13,19,21,2,2-tetramethyl- 28-dioxo-4-

β-azatricyclo [22.3.1.0] octacos-18-ene, as referred to in FR-900506, or by rapamycin. It is very common for patients to take too much of these medicines when they realize that they have forgotten to take their medicines at the prescribed time. This situation can cause serious side effects.

Another situation in which the compounds of formula I may be used to treat immunodepression is in vaccination. It is sometimes found that the antigen introduced into the body to acquire immunity to the disease acts as an immunosuppressive agent and thus the antibodies are not produced by the body and the immunity is not acquired. By introducing a compound of formula I into the body (for example through the vaccine) unwanted immunosuppression can be suppressed and immunity can be acquired.

In addition, the present invention further provides novel compounds of the formula I as previously defined, provided that X2 is i) when R 'is OH; represents H? R3 and R4. together represent a second carbon-carbon bridge between the carbon atoms to which they are attached

7 9 tram connected; R 2 is H; R 2 and X are each 0; and n is 2; thus R 3 is not 2-oxopropyl, 2,3-dihydroxypropyl or ethanalyl; ii) When R 2 represents OH; R 2 represents H; R 2 and R 3 together represent a second carbon-carbon bridge between the carbon atoms to which they are attached; R represents OH; R represents 3 OCH; X and Y are each 0; and n is 2; wherein R3 is not allyl or 1-hydroxyprop-2-enyl; and iii) when R 1 represents OH; R2 represents H; R5 and R6 together represent a second carbon-carbon bridge between the carbon atoms to which they are attached; R represents H; R represents OCH3? X and Y are each (H, OH); and n is 2; as well as R being non-allyl.

In the novel use or in the novel compounds, it is preferred that R is ethyl substituted by 0 or propyl substituted by 0.7

Desirably, R represents OH.

It is preferred that R2 is OH, preferably (S) -OH (i.e., having an absolute stereochemical configuration S, at its point of attachment to the molecule).

When R 3 includes an ester or amide group, it is preferred that the alcohol or the amide radical contains from 1 to 10 carbon atoms, for example the alcoholic moiety may be methanol.

The known compounds of formula I (as defined above) of WO 89/05304 include: 17-allyl-1,14,20-trihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) 1-methylvinyl] -2,3,25-dimethoxy-13,19,21,27-tetramethyl-11,28dioxa-4-asatricyclo [2.2.3.1.0,7] octanes-18-ene-2,3,10,16-tetrone , 17- (1-hydroxyprop-2-enyl) -1,14-20-trihydroxy-12β- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl- 25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.3.1.0 4,9] octacos-18-ene-2,3,10,16-tetraone. 17- (2,3-dihydroxypropyl) -1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy -13,19,21,29-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.3.1.04,8] octacos-18-ene-2,3,10,16-tetraone. 17-etenalyl-1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxyhexyhexyl) -1-methylvinyl] -23,25-dimethoxy-13,19,21,27- tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.04oct-cos-18-ene-2,3, 1Î ±, 16-tetraone. 17- (2-oxopropyl) -1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -β-methyl vinyl] -23,25-dimethoxy- 21, 27-te-

- Tramadol-11,28-dioxa-4-aza-bicyclo [2.2.1] octane-18-ene-2,3,10,16-tetraone. 17-allyl-1,12,14,16-tetrahydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methylvinyl] -2,3,25-dimethoxy- 21,27-tetramethyl-11,28-dioxa-4-azatricyclo [4.2.1] octane-18-ene-3,10-dione.

According to the present invention there is provided a process for the production of a novel compound of the formula I which comprises (a) the production of a compound of formula I in which R 3 represents propyl substituted by o, by oxidation of a corresponding compound in which R 3 represents allyl b) in the production of a compound of formula I which contains a vicinal diol group by oxidation of a carbon-carbon double bridge in a corresponding compound; C) in the production of a compound of formula I in which R is ethyl substituted by O by oxidative cleavage of a corresponding compound in which R3 represents 2,3-dihydroxypropyl; d) in the production of a compound of formula I in which R 3 represents methyl substituted by -CO 2 H or an ethyl group substituted by -CH 2 OH by oxidation of a corresponding compound in which R 3 represents ethanyl or propenalyl; e) in the production of a compound of formula I which contains two vicinal hydrogen atoms, by reduction of a corresponding compound containing a carbon-carbon double bridge? f) in the production of a compound of formula I in which X or Y represent (H, OH) by reduction of a corresponding compound wherein X, X or Y represent 0? g) in the production of a compound of formula I, in which R 3, R 4 and Y together with the carbon atoms to which they are attached represent a furanylmethyl ring substituted by the action of an acid on a corresponding compound in that 3 4. R 2 is 2-oxopropyl, R 2 is H and Y is 0? - 8 -

I

h) in the production of a compound of formula 1 in which R1 and R5 together represent an oxygen atom and R6 and R7 together represent a second carbon-carbon bridge between the carbon atoms to which they are attached by action of an acid on a corresponding compound in which R1 represents OH, R5 and R6 together represent a second carbon-carbon bridge between the carbon atoms to which they are attached, and R7 represents OH;

i) in the production of a compound of formula I in which R7 and R8 together represent an oxygen atom by the action of a dehydrating agent on a corresponding compound of formula I in which R7 represents OH and R8 represents OCH3; j) in the production of a compound of formula I, in which R 7 represents OH, through an allylic oxidation of a corresponding compound, wherein R 7 represents H; or (k) in the production of a compound of formula I, in which R 3 represents allyl substituted by hydroxy, through an allylic oxidation of a corresponding compound, wherein R 3 represents allyl.

The esters and amides of barbiturates which may be represented by R 3 may be produced according to conventional methods.

When desired or necessary, the hydroxy groups may be protected and deprotected using a conventional chemical protecting group Z bal as described in " Protective Groups in Organic Chemistry ", ed. 1 W P McOmie, Plenum Press (1973) and in " Protective Groups in Or " " Antic Synthesis ", T W Greene, Wiley Interscience 9

(1981) J. Further, European Patent Application No. 184162 describes the use of protecting groups in the macrocyclic compounds.

In process (a), suitable oxidizing agents include a palladium (II) halide, for example palladium (II) chloride, together with a copper halide, for example copper (I) chloride.

Suitable solvents include those which do not adversely affect the reaction, for example dimethylformamide (DMF) and water. The reaction is preferably carried out at a temperature in the range of 0 to 100 ° C, preferably at or near ambient temperature.

In process (b), suitable oxidizing agents include osmium tetroxide, potassium permanganate and iodine, together with silver acetate, osmium tetroxide is preferably used together with a regenerating agent such as 4-N-oxide -methylmorpholine. Suitable solvents include those which do not adversely affect the reaction, for example diethyl ether or tetrahydrofuran (THF). In the case of potassium permanganate, preference is given to alkaline aqueous solutions. The reaction is preferably carried out at a temperature of from 0 to 100 ° C, preferably at or near ambient temperature.

In process (c), the appropriate reagents preferably include a tetraacetate and phenylaldehyde acetate. Suitable solvents include those which do not adversely affect the reaction, for example benzene and glacial acetic acid. The reaction is preferably carried out at a temperature in the range of from 100 DEG C., preferably at or near ambient temperature.

In process (c), suitable oxidizing agents include sodium chloride together with hydrogen

sodium phosphate. Suitable solvents include those which do not adversely affect the reaction such as water. The reaction is preferably carried out at a temperature of from 0 to 100 ° C, preferably at or near ambient temperature.

In process (e), reduction of a catalytic mode using hydrogen can be carried out. Suitable catalytic agents include platinum catalytic agents (for example, black platinum) and palladium catalysts (for example palladium-on-carbon). Suitable solvents include those which do not adversely affect the reaction, for example methanol and ethanol. The reaction may be carried out at or near ambient temperature.

In process (f), suitable reducing agents include borane (for example as a borane-ammonium complex) and sodium borohydride. Suitable solvents include those which do not adversely affect the reaction, for example diethyl ether and dichloromethane. The reaction is preferably carried out at or near ambient temperature. Where desired or required, (H, OH) may be reoxidized to 0 by copper (II) acetate in acetic acid.

In processes (g) and (ft), suitable acids include p-toluenesulfonic acid. Suitable solvents include those which do not adversely affect the reaction, for example toluene. The reaction is preferably carried out at a temperature above room temperature, for example in a steam bath.

In process (i), suitable reagents include the Martin sulfurane reagent. Suitable solvents include those which do not adversely affect the reaction, for example dichloromethane. The reaction is preferably carried out at a temperature below ambient temperature,

for example at -30øC.

In processes (j) and (k), suitable reagents include S and O 2, preferably in the presence of the t-butyl hydrogen peroxide. Suitable solvents include dichloromethane and the reaction should be carried out at or near ambient temperature. The present invention further provides the use of novel compounds of formula I as pharmaceutical agents and a pharmaceutical composition containing such a compound in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

Of course, the dosage administered in the aforementioned therapeutic use will vary according to the compound used, the mode of administration, the desired treatment (for example parenteral or oral topical) and the referenced disease. However, in general, satisfactory results are obtained when administering the compounds at a dosage ranging from 0.1 to 200 mg per kg body weight of the animal. The total dosage indicated for man is in the range of from 1 mg to 100 mg and preferably from 10 mg to 500 mg which may be administered, for example twice a week, or in doses divided by 1 to 6 times a day or as a delayed release. Thus, dosage forms suitable for administration, for example by the esophageal route, should contain from 2 mg to 500 mg, and preferably from 1 mg to 500 mg of the compound, preferably in admixture with a solid, diluent, carrier or adjuvant or liquid, pharmaceutically acceptable salts.

Pharmaceutical compositions suitable for administration of the compounds of formula I (as defined above) should comprise (preferably 12

less than 80% and most preferably less than 50% by weight) of a compound of formula 1 '(as defined above) in combination with a pharmaceutically acceptable adjuvant, diluent or carrier. Examples of suitable adjuvants, diluents and carriers are the following: for tablets, capsules and dragees - microcrystalline cellulose, calcium phosphate, diatomaceous earth, a sugar, such as lactose, dextrose or mannitol, talc, stearic acid, starch, sodium and / or gelatin? for suppositories - natural or heavy oils or waxes? and for inhalation compositions - crude lactose. The compound of formula I (as defined above) should preferably have a shape having a mass median diameter of 0.01 to 10 microns. The compositions may further contain preservatives, stabilizers, wetting agents, , sweeteners, coloring and flavoring agents If desired, the compositions may be further elaborated as a delayed release. Preferred are those compositions which are to be taken via the esophageal route and which release their components into the body. gastrointestinal region.

In addition, it has further been discovered that the toxicity of the immunosuppressive compounds, in particular the immunosuppressive macrocyclic compounds including 12- (2-cyclohexyl-1-methylvinyl) -13,19,21,29- tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.0] octa-cos-18-ene (for example PR-9005-6) and rapamycin may be reduced by administering it together with a compound of formula I, as defined above.

Thus, according to the second aspect of the present invention, there is provided a pharmaceutical mixture comprising a compound of the formula I (as defined above) and an immunosuppressive compound. 13 -

Preferably, the largest proportion of the active ingredient in such a mixture consists of the compound of formula I ' for example, the compound of formula I may be present in a proportion greater than 10: 1 by weight, for example 99: 1. The present invention also provides a method for the treatment of immunosuppression or an immunosuppression disorder which comprises administering a therapeutically effective amount of a compound of formula I as defined above to a patient who is in such a situation.

The compounds of the formula I (as defined above) have the advantages, in relation to the compounds previously used in the above-mentioned therapeutic areas, that they are less toxic, more effective, have a longer action, have a broad field of activity are more potent, produce very small side effects, are absorbed more quickly or have other pharmaceutical properties.

The compounds of the formula I (as defined above) have a number of chiral centers and may exist in the form of a large stereoisomeric variety. The present invention may provide the use of all optical and stereoisomeric forms, as well as racemic mixtures and all optical and stereoisomeric forms of the novel compounds of formula I. The present invention may be illustrated by the following examples : 14 -

Example 1

17- (2-oxopropyl) -1-hydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methaninyl] -2,3,25-dimethoxy-13,19 Tetramethyl-11,28-dideoxy-4-azatricyclo [2.2.1.0,7] octacos-18-ene-2,3,10,16-tefracone a) 17- (2 -oxopropyl) -1-hydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -3-methylimino] < 23,25-dimethoxy-13,19,21,27-tetramethyl- 11,28-dioxa-4-azatricyclo [2.2.3.1] octane-14,18-diene-2,3,10,16-tetraone

J

To a solution of palladium (II) chloride (25 mg) and copper (I) chloride (50 mg) in DMF (dimethylformamide) (6 ml) and water (1 ml), which had been previously A solution of 17-allyl-1-hydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] 7,23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [

Octacose-14,18-diene-2,3,10,16-tetra-one (the compound of Example 4, 89/05304) (100 mg) in DMF (2 mg). The reaction was then stirred and oxygenated for 3 hours at room temperature, whereupon it was diluted with diethyl ether. The organic extract was then washed with dilute hydrochloric acid (1M) in brine and brine before being dried (MgSO 4), filtered and evaporated to give the title compound as an oil. b) 17- (2-oxopropyl) -1-hydroxy-1-2- [2- (4-hydroxy-3-methoxycyclohexyl-1-methylvinyl] -2,3,25-dimethoxy-13,19,21,27- tetramethyl-11.28-dioxa-4-azatricyclo-4,9- / 22.3.1.0 / octacos-18-ene-2,3,10,16, tetraone

The crude product from step (a) was redissolved in dry methanol and stirred with 10% Pd-on-carbon (10 mg) under a hydrogen atmosphere for 4 hours.

hours. The reaction mixture was then filtered, concentrated in vacuo and chromatographed on silica gel eluting with acetone / hexane (2: 3) to give the title compound as a foam (84%). mg).  € ƒâ € ƒâ € ƒMS (FAB): 889 (M +): Rb7 + 827 (M + Na +); 805 (m + h] +; 1 H NMR (CDCl 3): δ 211.1 (C 16): 207 (C 41); 196.3 (C2): 169.1 (C10); 166.1 (3); 138.7 (C19)? 132 (C31), 131 (C29), 121.8 (C18); 97.3 (GI); 84 (C34); 82.4 (Cl 2): 75.1 (C 23)? 56.2 (C9); 48.9 (0))? 47.6 (C17), 13.3 (C39)

Example 2 -

(Ethanalyl) in the form of an oil. The resulting compound was dissolved in butanol (25 ml) and 1-methyl-cyclohex-1-ene (6 ml) and sodium chloride solutions (0.8 g) were added portionwise over 10 minutes. g) and sodium hydrogen phosphate (0.81 g) both dissolved in 5 ml of water. After being stirred at room temperature for 0.5 hour, the mixture was diluted by the addition of ethyl acetate. After this operation, hydrochloric acid (2M) diluted in aqueous solution was added and the organic extracts dried (MgSO4), filtered and evaporated in vacuo to provide an oil. Chromatography on silica gel was then performed using a mixture of diethyl ether / methanol / acetic acid / 90% and then chromatographed on silica gel eluting with dichloromethane / methanol / acetic acid / 170: 9: 1 to give the title compound as a foam (402 mg, 32%). NMR (CDCl 3) δ 196.3 (C2) Î'177.7 (C41): 168.83 (C10): 164.6 (C3) Î'139.7 (C C 19), 132.7 (C 29), 129.3 (C 31), 120.6 (C 18), 97.3 (C 1), 84.2 (C 34), 70.5 (C 14), 9.8 (C 39 ).

Example 3 2- (14-Dihydroxy-12- [2- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -3,2-dimethoxy-13,19- 21,27-tetramethyl-4,9,11,22-dioxa-4-azatricyclo 22.3.1.0 7-octacos-18-ene-2,3,10,16-tetraone-7-yl ) -methylethanoic acid ester

To a solution of the title compound in Example 2 (50 mg), a solution of diazomethane with diethyl ether is added in dichloromethane (3 ml) at room temperature. The reaction mixture was then concentrated in vacuo and chromatographed on silica gel using acetone / hexane / 1: 2 as eluant to give the title compound as a solid. -

puma (48 mg). MS (FAB): 920 (M + H +); 858 [M + Na] +? 818 " M-OH7 +13 C NMR (CDCl3): 212.7 (C16); 196 (C2); 173 (C41); 168.8 (C10); 164.6 (C3) 139.5 (C19); 132.9 (C29); 129.1 (C31) 120.6 (C18); 97.2 (Cl); 84.2 (G34)? 70.8 (C14); 14.5 (C30); 9.7 (C39).

EXAMPLE 4 N-Marpholinamide of 2- (R) -1,4-dihydroxy-12Î ± - [2- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] 23, 25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricylole Γ 22.3.1, 49.9 Î'-octacos-18-ene-2,3,10, 16-tetraone-17-yl) -ethanoic acid

A solution of the compound of Example 2 (63.1 mg) morpholine (20 mg) diluted 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (2.5 g) was stirred at ambient temperature for 2.5 hours 28 mg) and 4-dimethylaminopyridine (2 mg) in dry dichloromethane (4 ml). After this operation, water was added and the reaction mixture was extracted with dichloromethane. After washing with dilute hydrochloric acid (1M) in aqueous solution and saturated sodium hydrogen carbonate solution, the dichloromethane extracts were filtered (MgSO4), filtered and evaporated in vacuo to give an oil. Chromatography on silica gel was then effected using hexane / acetone as eluent to give the title compound as a foam (58 mg, 84%). MS (FAB): 975 (M +); 891 [M + H] +? 873 / " M-OH 7 *. 13 * " ** C NMR (CDCl 3) δ 213.2 (C 16); 195.9 (C2); 170.1 (C41); 168.8 (C10); 164.8 (C3); 139.1 (C19); 133.2 (C29); 128.9 (C31); 121.5 (C18); 97.4 (Cl), * 84.2 (C34); 66.3 and 65.8 (morpholine CO); 101 (G39).

Example 5 N-Ethanolamide, 2- (1,14-dihydroxyl) -2- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23, 25-dimethoxy-13,19,22,22-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] 3,10,1-tetraone-17-yl) -ethanoic acid -1-®-4-methyl- n · m m m - · ~ i-ιτι ηι · ιΓ ~ '" The title compound was prepared from the compound of Example 2, following a similar method to that used in Example 4. MS (FAB): 950 (M + H +) +; 888 M + MaJ7 " 1 "; (M + H) + IR (+) C NMR (CDCl3) Î'; 61.2 (meth) ethanediamine); 44.0 (CN of ethanolamine) / 16.2 (G47); 15.8 (C43); 14.7 (C30); 9.7 (C39)

Example 6

2- (1,14-Dihydroxy-12- [2- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -3,2,25-dimethoxy- 19, 21, 27 " -49-tetramethyl-11,28-dioxa-4-azatricyclo / 22,3,1.0 * octacos-18-ene-2,3,10,16-tetraone-17- yl) -ethanoic acid methyl ester with glycine methyl ester The title compound was prepared from the compound of Example 2, following a similar method to that used in Example 4. MS (FAB): 977 (M-5Rb] + 915 (+ Na +) +; M + H +; 875 (M + H) +13 C NMR (CDCl3) .delta. 213 (G16)? 195, (C2) 171.7 (glycine carbonyl) t 52.4 (OCH3 of glycine); 9.9 (C39);

Example 7 2- (1,14-Dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,23,25- -dimethoxy-13,19,21,29-tetramethyl-11,28-dioxa-4-azatricyclo 19-

- / [. The title compound was prepared from the compound of Example 2, except that the title compound was prepared from the title compound as a white solid.1H NMR (DMSO-d6, following a similar method to that used in Example 4. MS (FAB): 973 (M + 1) +; 911 Vi + Na2 +; 1 H-NMR (CDCl 3) δ 213/4 (ClO); 196 (C2); 169/4 (CIO); 168.6 (C41) / 164.7 (C3); 138.8 (CL9)? 133.3 (G29)

Example 8 N-benzylamide of 2- (1-hydroxy-12-Z-2- [2- (4-hydroxy-3-methoxycyclohexyl) dimethyl-13,19,21,21,29-tetramethyl-1 H -indole-4-azatricyclo [22.3.1.0 4/9] octacos-18-ene-2,3 , 10,16-tetraone-17-yl) ethane The title compound was prepared from the compound of Example 2, following a method similar to that used in Example 4. MS (FAB): 995% 933, " M + Na7 +; 911 [M + H] +; 1 H-NMR (CDCl 3, δ: 213.3 (C 16), 196.1 (C 2), 171.6 (C 41), 128.9 (C), 128 (C) 4 (G18), 97.5 (Cl), 84.3 (C34), 10.2 (C39)

Example 9 N-Butylamide 2- (1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] 23, 25-dimethoxy-13β, 21β, 21β, 22β, 13β-tetrahydro-11β-dioxa- [4-azatricyclo] , 18-ene-octacos-18-ene-2,3,10,16-tetraone-17-yl) -acetic acid The title compound was prepared from the compound of Example 2, following a method similar to that used in Example 4.

 € ƒâ € ƒâ € ƒMS (FAB) Î'961 M-fBb / 8,899 M + Na + 877 Z + M + H7 +? 859 / Î ± ηη7 + 13C rMN (GDC1) Î »213.4 (C16) / 196.1 (C2) 171.7 (C41)

Example 10 2- (1,14-Dihydroxy- [2- (2-oxo-3-methylethyl) cyclohexyl) -1-methylvinyl] -benzoic acid, dimethyloxy-13,19,21,2,2-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] octane-2,3,16,16- tetraone-17-yl) -ethanoic acid

A solution of the compound of Example 2 (51 mg) 2-chloro-1-methylpyridinium tosylate (33.4 mg), triethylamine (29 μM) and p-cyanophenol (10.2 mg) in THF (2 ml). Then, hydrochloric acid was added in aqueous solution and the organic extracts were dried (MgSO4) (after being washed with a saturated aqueous sodium hydrogen carbonate solution), filtered in vacuo to give an oil.

Chromatography on silica gel was then performed using dichloromethane / methanol Z 39: 1 to give the title compound as a foam (24.9 mg, 43%). MS (FAB): 1007 Z "M * RbJ7 +; NMR (CDCl3) δ 212.4 (C16), 196 (C2), 170.1 (C41)? 168.8 (€ 10)? 164.4 (C3), 153.8 (OC), 140.4 (C19), 133.6 (C), 132.7 (C29), 129.1 (C31), 122.6 (Car), 120.1 (C18), 118.2 (CN of p-cyanophenol), 10g, 7 (E arCN), 97 (Cl), 84.1 (C 34), 14.4 (C 30), 9.9 (C C39)

Example 11 2- (1,14-Dihydroxy-2- [2- (4-hydroxy-3-methoxy-6-hydroxyethyl) -1-methylvinyl] -dimethoxy-13,21

Tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] octane-18-ene-2,3,10,16-tetraone- 17-yl) -ethanone The title compound was prepared from the compound of Example 2, following a similar method to that used in Example 10. MS (FAB): MS (Rf) + 920 (M + Na + ; 898 [Î ±] +7.7807 [M +] + 13C NMR (GDG13) Î »: 212.4 (C16); 195.9 (C2), 171.1 (C41); 168.8 (C10); 164.5 (C3); 150.5 (OC); 140 (C19). Ο

2- (4-Hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,3,25-dimethoxy-2- (1,1-dimethylethyl) 13,19,21,21,7-tetramethyl-11,28-dioxa-4-azatopyran-4,9-v / 22.3.1.0 'octacos-18-ene-2,3,10,16-tetraone-17 -yl) -ethanoic acid The title compound was prepared from the compound of Example 2, following a similar method to that used in Example 10. MS (FAB): m + R @ 3 +; NMR (CDCl3): 212.6 (C16), 196.2 (C2), 170.3 (C41), 169 (C10), 164.7 (C3), 155 , 4 (OC), 145.5 (ct)

EXAMPLE 13 17-Allyl-14,14-trihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl-1-methylvinyl] -23,25-dimethoxy-13,19,21, 27-tetramethyl-11,11-dioxa-4-a, zatricyclo 22.3.1.04.99 octacos-18-ene-2,3,10-trione A was added to a solution of FR-900506 (100 mg) with ether / dichloromethane-diethyl ether an excess borane-ammonia complex (150 mg) After stirring for 2 hours,

at room temperature, hydrochloric acid (1M) was added in aqueous solution, dried (NaSO 4) and the crude extract filtered, and evaporated in vacuo to provide the crude 1,1,2,14,16- tetrahydroxyacetic acid as an oil. The resulting compound was placed in acetic acid and copper (II) acetate (100 mg) was added. After heating on a steam bath for 10 minutes, the reaction mixture was cooled to room temperature and water was added. The reaction mixture was then extracted with ethyl acetate and the organic extracts, then washed with saturated aqueous sodium hydrogen carbonate, dried (MgSO4), filtered and evaporated in vacuo to give to obtain an oil.

Chromatography was then carried out on silica gel using acetone / hexane (1: 1) as eluant to give the title compound (30 mg) in a stereochemical C16 -C16 configuration, followed by the title compound ( 24 mg) having a stereochemical S-configuration at C16. (16R) -Etereoisomer δ (FAB) 890 M + Rb7 * 13 C NMR (CDCl3) δ: 199.4 (C2) 169.1 (C10) (16S) -Estereoisomer) MS (FAB) 890 (M + H +) +; 828 [M-i-Na] 7 * 806 [M + H] +? 1 H-NMR (CDCl 3) δ (CDCl 3) δ (CDCl 3) 1,14-dihydroxy- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -26,28-dimethoxy- 13,18,22,24.30-pentamethyl-11,17β-trioxa-4-azatriazine [Î ±] 25, 3, 01, 04, 9, 16, 20, 16- 20 (20), 18.21 -triene-2,3,10-trione

A solution of 1,14-dihydroxy-12 - [[2 - [[[1,2,3]

- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,2-dimethoxy-17- (2-oxopropyl) -13,19,21,22-tetramethyl-11,28-dioxa-4 (the compound of Example 29, WO 89/05304) (100 mg) in dichloromethane dry dichloromethane (25 ml) containing p-toluenesulfonic acid (5 mg). The volatile products were then removed in vacuo and the residue chromatographed on silica gel eluting with acetone / hexane / 3: 2 as eluant to give the title compound (19 g) as a white solid. foam.  € ƒâ € ƒâ € ƒMS (FAB): 887 (M + H + +) + 325 + 785

13 C NMR (CDCl 3, δ 195.9 (C 2), 168.9 (CDCl 3), 164.9 (C 3), 150.8 (C 16), 148.3 (C 18) 106.3 (C19): 97 (Cl): 84.1 (C37): 8.5 (C42)

EXAMPLE 15 1.14-Hydroxy - [2- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,3,25-dimethoxy] -17-propanalyl-13,19,21 tetraethyl-11,28-dioxa-4-azatricyclo [22.3.1.0] octacos-18-ene-2,3,10,16-tetraone

A solution of FR-900506 (3 g) in DMF (90 ml) and water (15 ml) containing palladium (II) chloride (0.4 g) and copper (I) chloride (1.8 g ) at room temperature while bubbling air through the reaction mixture for 2 hours. The reaction mixture was then diluted in diethyl ether and the organic extract was dried (MgSO4) (after washing with hydrochloric acid ( 1M) diluted in aqueous solution, water and brine) filtered and evaporated in vacuo to provide an oil. Chromatography was performed on silica gel, eluting with hexane in an increasing gradient of acetone to give the title compound as a foam (30 mg). 24

NMR (CDCl 3) δ 213.1 (C16), 202.1 (G42), 186.3 (G2), 169 (C10), 164.7 (M + H) +. C3), 139.8 (C19), 13.26 (C29)

Example 16 17-Allyl-1 '' - hydroxy-12 '- [2' - [ 2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,23,25-dimethoxy] 13,19. Tetramethyl-11,11,14,14,14-trioxa-4-azatholol / 22, 3,1,1 ', 10-monacos-19-ene-2. 3,10,16-tetraone

A sample of 17-allyl-14,16-trihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methyl- dimethoxy-13,19,21,22-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] oct-18-ene-2,3,10,16-tetra (as prepared in Example 13 of WO 89/05304, 60 mg) over 10 minutes in toluene (10 ml) containing p-toluenesulfonic acid. Evaporation of the solvent in vacuo and chromatography on silica gel gave the title compound as an oil (30 mg). NMR (CDCl 3) δ: (single rotamer) 210.22 (C16 (Î'195.96 (C2); 168.96 (C10); 165.24 (C3); 135.92 (20) 802.47 886.2 (M + H +) +

Example 17 17-propyl-1-hydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -25-methoxy-13,19. 21. 27-tetramethyl-Î ±, Î ±, β, β- [Î ±] -4- [Î ±] -1-oxazin-4-one. . * .0 (nonas-18-ene-2,3,10,16-tetraone

A sample of 17-allyl-1,14,20-trihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25- dimethoxy-13,19,21,27 tetramethyl-11,28-dioxa-4-aza-trichloroethyl, 3,1,0, 4-octacos-18-ene. 2,3,10,16-tetraone (as prepared in Example 13 of V70 89/05304, 121 mg) in dry dichloromethane (10 ml) and additionally,

was added a Martin stilfuran reagent at ~ 30 ° C. After heating to 0 ° C, the volatiles were removed in vacuo and the residue was chromatographed on silica gel to give a foam. To a solution of the product in dry methanol (5 ml), 10% Pd-emarbonium (10 mg) was added to the solution and the resulting suspension was stirred at room temperature for 7 hours under an atmosphere of hydrogen. the volatiles were removed in vacuo and the residues were purified by column chromatography on silica gel to give the title compound as an oil (19 mg). MS (FAB): 770.76 [M + H] + = 792.74 [M + Na] +;  € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒâ € ƒ854.44 (M-H) +: 196.11 (C2), * 168.96 (C10); 165.21 (C3); 146.16 (C14) 56.47 (OCH3); 55.95 (C H 3); 51.37 (C17)

Example 18

17-alll-1-hydroxy-2- [2 (4-hydroxy-3-methoxy-cyclohexyl-1-methylvinyl] -25-dimethoxy-13,19,21,27,7-tetramethyl -11,28-29-trioxa-4-azatricyclo [22.3.1.1.02.0 nonacosa-14,18-diene-2,3,10,16-tetraone

A sample of 17-allyl-14,20-trihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy-13,19 , 21,27-tetramethyl-11,28-dioxa-4-azatra-cyclo [2.2.3.1.0] octacos-18-ene-2,3,10,16-tetraone (as prepared in Example 13 of WO 89/05304, 80 mg) was dissolved in dry dichloromethane (5 ml) and a solution of Martin's sulfurane reagent was added portionwise at -30 ° C until all starting material had disappeared (30 minutes). Then, the reaction mixture was diluted with ethyl acetate, washed with water and then brine. The organic extracts were dried (MgSO4), filtered and evaporated in vacuo to provide an oil. Chromatography on silica gel provided the title compound

The title compound was obtained as an oil (20 mg). NMR (CDCl3) C? * (mixture of rotamers) 212.05, 211.04 (Cl 6); 197.47, 194.95 (C2); 169.24, 169.09 (C10). MS (FAB): 775.11 (M + H) + = 797.09 [M + Na] +; £ 858.79 M-> b__7 +

Example 19 12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -23,25-dimethoxy-17-propyl-13,19,21,22-tetramethyl- 15-trihydroxy-11,28-dioxa-4-azatricyclo [22.3.1.0,7-octacos-18-ene-2,3,10,16-tetraone

A sample as an oil of 1-hydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,3,25-dimethoxy-17-propyl Methyl-11,28-dioxa-4-azatricyclo [22.3.1.0] octacosa-14,18-diene-2,3,10,16- tetraone (the compound referred to in Example 11 of WO 89/05304, 510 mg) and in dry tetrahydrofuran (12 ml) was added N-methyl morpholine N-oxide (0.3 g) and osmium tetroxide 3 ml) of 4% aqueous solution. Then, it was stirred at room temperature for one hour, and solid sodium metabisulfite (1 g) was added, followed by celite, after 5 minutes to obtain a thick mixture. The reaction mixture was then diluted with ethyl acetate and filtered. The organic extract was dried (magnesium sulfate), washed with saturated aqueous sodium bicarbonate solution and brine, filtered and concentrated in vacuo to provide an oil. Column chromatography on silica gel, using acetone / hexane / 2: 5-7 as the eluent gave two isomers of the title compound which differed by their stereochemical configuration at G14 and / or C15.

Isomer 1 (180 mg) 27

Claims (1)

1 * 3 -.  € ƒâ € ƒâ € ƒ1 C NMR (for the major rotamer) 211.82 (C16); 196.03 (C2) f 169.10 (C10); 164.26 (C3); 139.28 (C19): MS: 823 (M + Na +) + 906 / Fi + RbJ * Isomer 2 (32 mg) Î'NMR (for the main rotamer): 212.99 (C16): 194.39 (C2 ); 169.60 (C10), 164.85 (C3); 138.33 (C19) EMI 823 / " μ + Η_7 +; Example 20 In the mixed lymphocyte reaction (MLR) (described in WO 89/05304, Example A), it was found that the compound 17- (2-oxopropyl) -N-methyl- 1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl] -2,2-dimethoxy-13,19,22,21-tetramethyl- 11, 28-dioxa-4-azatricyclo [22.3.1.0] octacos-18-ene-2,3,16,16, had a pA2 value of 8.3 for the antagonist compound ER-900506. This means that a concentration of 5x10 4 of the antagonist compound is required to occupy 50% of the available receptor sites in the screening, according to which it competes with FR-900506. A process for the preparation of a compound of formula I wherein R1 and R2 independently represent H or OH, or together may represent a second carbon-carbon bond between the carbon atoms to which they are attached; represents methyl optionally substituted by -CO-α or an ester or amide thereof; ethyl optionally substituted with 0, OH or -CD2H or an ester or amide thereof; propyl optionally substituted by OH or O; or ally optionally substituted by OH; represents H; R 2 and R 3 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H or OH; 29 - R.8 R 9 and Y 9 represents OH or OCH 4 represents 0 or (H, 01-1); represents 0 or (¾ OH); and n is 1 or 2; in addition to the above meanings R1 and R23 together may represent an oxygen atom and in which case R6 and R6 together represent a carbon-carbon second bond between the carbon atoms to which they are attached; R 2 and R 3 may together represent an oxygen atom; and R3 and R4 together with the carbon atoms to which they are attached may represent a furanyl ring substituted by methyl; provided that 23 i) when R represents H; Râ 'is methyl, ethyl, propyl or allyl; R 2 and R 3 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 2 is 0 and Y is 0; then R 2 is OH; and 1 2 5 when R x is OH; R represents H? R 2 and R 3 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 2 is H; R 2 is OCH 2; each X and Y represents O? and n is 2; then R 2 does not represent 2-oxopropyl, 2,3-dihydroxypropyl or ethynyl; Ii) when n is 1, then R 2 is not methyl or ethyl; in the condition of 3 i) Ii) when R 2 is OH, R 2 is H; R3 and R6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached R3 is OH; R 2 is OCH 3; each X and Έ is 0; and n is 2? then R3 is not allyl or 1-hydroxy-prop-2-enyl; and iii) when R is OH? R represents H? R3 and R6 together represent a second carbon-carbon bond between the carbon atoms to which they are so attached; R represents H? R4 is OCH3; each X and Y is (H, OH)? and n is 2? Then R does not represent allyl. 1 2 Iv) in the case where the radicals R and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached? the radi-56 moieties R and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached; the radical R 2 represents hydrogen atom; the radical R 2 is the group 0 CH 3 - X is the oxygen atom; Y is the group (H, OH); and n is the integer 2? then the 3 r radical does not represent the allyl group? and (v) in case the radical R 1 is OH; the radical R4 represents the hydrogen atom; the radicals R and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached; the radical R represents the hydrogen atom; the radical R 2 represents the group OCH 3; X is the group (H, OH); Y is the oxygen atom? and n is the integer 2? then the radical does not represent the ethanalyl group? A compound of formula I wherein R is propyl substituted by 0 is produced by oxidation of a corresponding compound in which R is allyl; there is produced a compound of formula I which contains an adjacent diol by oxidation of a carbon-carbon double bond to a corresponding compound; there is produced a compound of formula I in which R 2 represents ethyl substituted by O by oxidative cleavage of a corresponding compound in which R 2 represents 2,3-dihydroxy-propyl; there is produced a compound of formula I in which R 3 represents methyl substituted by -CH 2 H or substituted ethyl-CO 2 H by oxidation of a corresponding compound in which R represents ethanalyl or propanalyl; there is produced a compound of formula I which contains two adjacent hydrogen atoms by reduction of a corresponding compound containing a carbon-carbon double bond; there is produced a compound of formula I in which X or Y represents (H, OH) by reduction of a corresponding compound in which X and Y represents O; there is produced a compound of formula I in which R 1 and R 2 and Y together with the carbon atoms to which they are attached represent a furanyl ring substituted by methyl, the acid in a corresponding compound in which 34 is 2-oxopropyl, R3 is H and Y is O; h) i) j) k) a compound of formula I is produced in which R 2 and R 3 together represent an oxygen atom and R 2 and R 3 together represent a second carbon-carbon bond between the carbon atoms which are linked by the action of acid on a responding compound in which R 2 represents OH, R 2 and R 3 together represent a second carbon-carbon bond between the carbon atoms to which they are bonded and R 2 is OH; 7, a compound of formula I in which R 1 and R 2 together represent an oxygen atom is produced by the action of a dehydrating agent on a corresponding compound of formula I in which R 2 is OH and R 2 is OCH 3, ; 7 a compound of formula I in which R is OH is produced by allylic oxidation of a corresponding compound in which R represents H; or a compound of formula I in which R represents a hydroxy-substituted allyl group is produced by allyl oxidation of a corresponding compound in which R represents alkoxy. A process according to claim 31, wherein R is ethyl substituted by O or propyl substituted by O, A process as claimed in claim 7, wherein R is OH ' 4. A process as claimed in claim 21, wherein R is OH? 17-allyl-1,14,20-trihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -piperazin-1-yl] -amide The compound according to claim 1, 7-methyl-vinyl 7-23, 25-dimethoxy-13,19,21-27-tetramethyl-11,28-dioxa-4-aza-tricyclo [23.3.1.0] octacos-18- 2, 3,10,16-tetra-one. 17- (1-hydroxy-prop-2-enyl) -1,14,20-trihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl- Dimethoxy-13,19,21,27,29-tetra-methyl-11,28-dioxa-4-aza-tricyclo [2.2.2] octane-18-ene- 2,3,10,16-tetra-one. 17- (2,3-dihydroxy-propyl) -1,14-dihydroxy-12-7- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl- , 25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [23.3.1.0 octacos-18-ene-2,3,10,16-tetraone. 17-ethanalyl-1,14-dihydroxy-12 - ([2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,3,25-dimethoxy-13,19 , 21,27-tetramethyl-4-9,28-dioxa-4-azatricyclo [23.3.1.0] octacos-18-one-2,3,10,16-tetraone. 17- (2-oxopropyl) -1,14-dihydroxy-12-7-2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -23,25-dimethoxy-13 Tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] octane-18-ene-2,3,10,16-tetraone, or 17-allyl-1 1,2,4,16-tetrahydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -23,25-dimethoxy- 19,21,27-hexahydro-11,28-dioxa-4-azatricyclo [2.2.1] octane-18-ene-3,10-dione 2. A process according to claim 1, wherein the following compounds of formula 17- (2-oxopropyl) -1-hydroxy-12- [2- (4-hydroxy-3-methoxy- -hexyl) -1-methyl-vinyl] -2,3,25-dimethoxy-13,19,21,27-tetramethyl-9,11-dioxa-4-azatricyclo [2.2.3.1.0 Octacosa-14,18-diene-2,3,10,16-tetraone. 17- (2-oxopropyl) -1-hydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -23,25-dimethoxy-13,19,21 , 27-tetramethyl-9- [11,28-dioxa-4-azatricyclo [22.3.1.0 '/ octacos-18-ene-2,3,10,16-tetraone, 2- - (1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl-1-methylvinyl] -2,3,25-dimethoxy-13,19,21,2,7-tetramethyl- Dihydro-4-azatricyclo [2.2.1] octane-18-ene-2,3,10,16-tetraone-17-yl) ethanoic acid , 2- (4-hydroxy-12-methoxy-cyclohexyl) -1-methyl-vinyl-2S-dimethoxy-2- Tetramethyl-11,28-dioxa-4-azatricyclo2.2.3.1.0 (octanes-18-ene-2,3,10,16-tetraone-17-yl) ethanoic acid , 35 - 2- (1,14-Dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,3,25-dimethoxy- tetramethyl-11,28-dioxa-4-azatricyclo [2.2.3] octacos-18-ene-2,3,10,16-tetraone-17-yl) 2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,3,25-tetrahydroisoquinolinecarboxylic acid N-ethanedioate, 2- (1,14-dihydroxy- dimethoxy-13,19,21,2,2-tetramethyl-11,28-dioxa-4-azatricyclo 22.3.1.04,9] octacos-18-ene-2,3,10,16-tetraone-17-yl) ethanoic acid , 2- (1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl) -23,25-dimethoxy- -13,19,21,28-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.3] octane-18-ene-2,3,10,16-tetraone-17- yl) -ethanoic acid methyl ester with glycine methyl ester, 2- (1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) dimethyl-13,19,21,23-tetramethyl-11,28-dioxa-4-azeza [c] [2] benzazepine-2-carboxylic acid [ 3,10,16-tetraone-17-yl) -ethanoic acid, 2- (1,14-Dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -23,25- dimethoxy-4,13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.0-octacos-18-ene-2,3, o, 16-tetraone-17-yl) ethanoic acid , 2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl-7- (1,1-dimethylethyl) Dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [2.2.1] octane-18-ene-2,3,10,16 17-yl) -ethanoic acid, 2- (1,14-dihydroxy-12-7- (4-hydroxy-3-methoxy-cyclohexyl) -1- methyl-vinyl] -2,3,25-dimethoxy-4,13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.0] -octacos-18-ene-2,3,10,16- tetraone-17-yl) -ethanoic acid. 36 2- (4-Hydroxy-3-methoxy-cyclobenzyl) -1-methyl-vinyl] -23,25-dimethoxy-2- (1,14-dihydroxy- Tetramethyl-11,28-dioxa-4-azatricyclo [2.2.3.1.0 4,9] -octacos-18-ene-2,3,10,16-tetraone-17-yl) 2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,2,3,4-tetrahydroisoquinoline-2-carboxylic acid ester p-nitrophenyl ester methoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclopent-9-ene-22.3.1.0 '- octacos-18-ene-2,3,4,5- 17-allyl-1,14,16-trihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -piperidine] dimethyl-13,19,21,22-tetramethyl-δ-11,28-dioxa-4-azatricyclo [2.2.3.1.0] octacos-18-ene -2,4,10-trione, 1,14-dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -26,28-dimethoxy-13 , 18, 22, 24/30-penta-methyl-11,17,13-trioxa-4-azatricyclo [25.3.1.0 [(4-chlorophenyl) 21, triene-2,3,10-trione. 14-Dihydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,3,25-dideoxy-17-propanalyl-13,19,21 , 17-allyl-17-en-2,3,10,16-tetraone, 17-allyl-1-azabicyclo [2.2.1] octane- hydroxy-12- [2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -2,3,25-dimethoxy-13,19,21,2,2-tetramethyl- 3-cyclohexyl-4-azatetracyclo [2.2.3.1.1, 7-nonoxo-2,3,10,16-tetraone, 17-propyl-1-hydroxy-1,2- 2- (4-hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl) -25-methoxy-13,19,21,29-tetramethyl-11,28-29-29 17-allyl-1-hydroxy-12- [2- (4-azabicyclo [3.2.1] octane- hydroxy-3-methoxy-cyclohexyl) -1-methyl-vinyl] -25-methoxy-13,19,21,23-tetramethyl-11,28,29-trioxa-4-azatetracyclo [ (4-hydroxy-3-methoxy-cyclohexyl) -7-methyl-1,2,3,4-tetrahydronaphthalene, -23,25-dimethoxy-17-propyl-13,19,21,27-tetramethyl-1,14,15-trihydroxy-11,28-dioxa-4-azatricyclo A process for the preparation of a pharmaceutical composition which comprises as active ingredient a compound of formula I, when prepared according to the preceding claims, in association with a pharmaceutically acceptable adjuvant, diluent or carrier. 7, characterized in that an immunosuppressive compound is additionally incorporated. The applicant claims the priorities of the UK applications lodged on 14 and 15 September 1989 under Nos 89/20849 and 89/20985 and 22 March 1990 under Nos 90/06449, 8 June 1990 under the N2 90/12795 and 6 July 1990, under Nos. 90/14959. Lisbon, September 13, 1990