PT98175A - Process for the preparation of tricyclic compounds containing hetero atoms and intermediate products - Google Patents

Abstract

This invention relates to a process for the preparation of tricyclic compounds containing hetero atoms, namely macrolides, in particular a process for the preparation of derivatives of 12-(2'-cyclopentylvinyl)-11,28-dioxa-4- azatricyclo-[22.3.1.0 4,9]octacosene-18 or 12-(2'- cyclopentylvinyl)-11,28-dioxa-4-azatricyclo-[22.3.1.0 4,9]octacosadiene-14,18 and derivatives of 11-(2'- cyclopentylvinyl)-10,27-dioxa-4-azatricyclo-[21.3.1.0 4,8]heptacosene-17 or 11-(2'-cyclopentylvinyl)-10,27- dioxa-4-azatricyclo-[21.3.1.0 4,8] heptacosadiene-13,17, in particular compounds of formula I <IMAGE> in which the substituents have the various meanings mentioned in the claims, which includes a reaction for contracting the ring accompanied by preparation of the appropriate derivative. These have a useful pharmacological activity as immunosuppressive and antiproliferative and anti-inflammatory agents and as chemotheraputic agents for reversing drug resistance.

Description

(I.e.

The present invention relates to a process for the preparation of tricyclic compounds containing heteroatoms. It refers to the field of macrolides.

Referring in particular to a process for the preparation of a 12- (2'-cyclopentylvinyl) -1,28-dioxa-4-azatricyclo [22.3.1.0 3-octacos-18-ene, or 12- (2 '-cycle-

pentylvinyl) -11,28-dioxa-4-azatricyclo E22.3.1.09-9] -octacos-14,18-diene, or 11- (2'-cyclopentylvinyl) -10,27-dioxa- 4-azatricyclo [21.3.1.0 11,8] -heptacos-17-ene, or 11- (21-cyclopentylvinyl) -10,27-dioxa-4-azatricyclo [21.3.1. 0 *] -heptacos-13,17-diene, with the proviso that its cyclopentyl moiety is other than 3-formylcyclopentyl, hereinafter abbreviated as &quot; a compound of the invention &quot;.

A preferred compound of the invention is: a 17-alkyl (or 17-alkenyl) -1-hydroxy-14-oxy-12- (2'-cyanopentyl-1'-methyl-trans-vinyl) -23,25-dimethoxy - 13,19,21,27-tetra-estr-11,28-dioxa-4-azatricyclo [22.3.1.0 11,9 3-trans-octa-cosene-18-2,3,10,16-tetrone, or a derivative of 17-alkyl (or 17-alkenyl) -1-hydroxy-12- (2'-cyclopentyl-1'-methyl-trans-vinyl) -23,25-dimethoxy-13,19,21,27-tetramethyl- , 28-dioxa-4-azatricyclo [22.3.1.0 4,9 3-trans-octacosadiene-14,18- (or trans-18-octacho-sene) -2,3,10,16-tetrone, with condition that its part of cyclopentyl is different from 3-formyl-cyclopentyl. (I.e.

Even more preferred is: a 17β-alkyl (or 17Î ± -alkenyl) -1- (Î ± -hydroxy-14Î ± -oxy-12β- [2β- (cyclopent-1 (R) -yl) -1-methyl 1-trans-vinyl-23-dioxa-4-azatricyclo [2.2.0] octane-18-ene-17- (17Î ± -alkenyl) -1β-hydroxy-12- [2 '- (cyclopent-1-yl) (R) -1 ') - 1'-methyl-trans-amino-23β, 25β-dimethoxy-13α-, 9,21α, 27β-tetramethyl-11,28-dioxa-4-azatricyclo - [22.3.1.03-trans-octacosa-diene-17,18- (or-trans-18-octane-no) -9H, 24H-2,3,10,16-tetrone, with the proviso that their cyclopentyl moiety is other than 3-formylcyclopentyl. The cyclopentyl moiety is preferably 4

monocyclic, preferably at the 3-position. The tricyclic ring structure is preferably substituted as is known from the literature, e.g., from EP 184162, for analogous cyclohexyl tricyclic compounds. The compound of the present invention may be obtained by a process which comprises subjecting a 12- (2'-cyclohexylvinyl) -1,2,8-dioxa-4-azatricyclo [22.3.1, Îμ] octacoseno-18 or -14,18-diene, or a derivative of 11- (2'-cyclohexylvinyl) -10,27-dioxa-4-azatricyclo [22.3. 1.0-heptacoseno-n- or -13,17-diene, for ring contraction and appropriately deriving the resulting cyclopentyl compound. The invention further includes the use of this resulting cyclopentyl compound as an intermediate, e.g., in the preparation of a compound of the invention. The process of the invention is carried out in an analogous manner to known processes. Ring shrinkage is, e.g., effected by treating a corresponding derivative substituted on the cyclohexyl ring by a cyano group, with an acid having a non-nucleophilic anion, such as hydrofluoric acid. Preferably, an inert solvent such as

acetonitrile. The reaction is effected, e.g., in an analogous manner to that described in EP 427680. Protecting groups which may be conveniently present may be removed simultaneously or protection groups may be introduced in a conventional manner. The temperature is preferably about room temperature. Subsequent derivatization is also carried out in conventional manner, e.g., according, or analogously, to the methods described below.

The compounds of the invention may be isolated and purified from the reaction mixture in a conventional manner.

The designations and numbering of the carbon atoms used above are in accordance with the designations and numbering in EP 184162 for cyclohexyl analog compounds. However, various other designations and numbering are used in the literature. The numbering of the carbon atoms below used is in accordance with the numbering for compound FK506, published in H. Tanaka et al., J. Am. Chem. Soc. 109 (1987) 5031.

The present invention relates in particular to compounds of formula I

on what

R 1 is methoxymethyl; optionally protected hydroxymethyl; acyloxymethyl, wherein the acyl part optionally contains either a dimethylamino group which may be optionally quaternized, or a carboxy group which may be optionally esterified, or one or more amino groups and / or hydroxy groups which may optionally be protected; or aminooxyalkoxymethyl; or is optionally protected methoxy, formyloxy or hydroxy and there is a single bond between positions 23 and 24 or Rz is absent and there is a double bond between positions 23 and 24;

and R 3 is methyl, ethyl, n-propyl or allyl; in free form and, if such forms exist, in salt form, hereinafter referred to briefly as &quot; the particular compounds of the invention &quot;.

As shown in formula I, the carbon at the 32-position is asymmetrically substituted; Râ,, may thus be attached in the configuration or 9 to that carbon atom and the compounds of formula I may exist as the corresponding corresponding isomers or in the diastereoisomeric mixture form thereof.

A subgroup of particular compounds of the invention is that of the compounds Ipl, ie the compounds of formula I wherein * is hydroxymethyl, or R2 is optionally protected hydroxy and there is a single bond between positions 23 and 24, or Rz is absent and there is a double bond between positions 23 and 24; and R 3 is as defined in formula I.

A further subgroup of particular compounds of the invention is that of the compounds Ip 2, i.e. the compounds of formula I, wherein t

R 1 is optionally protected hydroxymethyl;

Rz is as defined for compounds Ipl; and R 3 is as defined in formula I; in free form and, when such forms exist, in salt form.

A further sub-group of particular compounds of the invention is that of the compounds Ip 3, i.e. the compounds of formula I in which:

R1 is from the acyloxymethyl formyloxymethyl group; or Rz is optionally protected formyloxy or hydroxy and there is a single bond between positions 23 and 24, or is absent and there is a double bond between positions 23 and 24; and R 3 is as defined in formula I, in the free form and, when such forms exist, in salt form.

Another subgroup of particular compounds of the invention is that of the compounds Ip4, i.e. the compounds of formula I, wherein 7

R4 is acyloxymethyl wherein the acyl moiety optionally contains either a dimethylamino group, which may be optionally quaternized, or one or more amino and / or hydroxy groups which may be optionally protected; R-z. is as defined above for compounds Ipl; and R 3 is as defined above in formula I; in free form and, when such forms exist, in salt form.

Another subgroup of particular compounds of the invention is that of the compounds Ip 5, i.e. the compounds of formula I wherein:

R1 is aminooxyalloxymethyl;

Rz is as defined above for compounds Ipl; and R 3 is as defined above for formula I; in free form and, when such forms exist, in salt form.

Another sub-group of particular compounds of the invention is that of the compounds Ip6, ie those compounds of formula I wherein: or R3 is methoxymethyl and R3 is methoxy or optionally protected hydroxy and there is a single bond between positions 23 and 24 , or is absent and there is a double bond between positions 23 and 24; or R1 is optionally protected hydroxymethyl and Rz. is methoxy; and R 3 is as defined in formula I; in free form, and when such forms exist, in salt form.

In an appropriate acyloxymethyl group, the acyl moiety preferably has a total of 1 to 20, preferably 1 to 5 carbon atoms, e.g. formyl, acetyl, isobutanoyl, benzoyl or pivaloyl. It is preferably formyl.

Suitable protecting groups for hydroxy and hydroxymethyl are conventional hydroxy protecting groups, such as tert-butoxycarbonyl, or trialkylsilyl, preferably tert-butyldimethylsilyl.

Hydroxymethyl or optionally protected hydroxy, as defined above in formula I for R1 and R2, is not to be understood as including a group R3 or R4 which is specified elsewhere in the definitions for formula I, such as acyloxymethyl , aminooxyalloxymethyl, formyloxy or methoxy. The preferred amino is preferably amino protected by a conventional amino protecting group, such as

bezyloxycarbonyl, or trialkylsilyl; is especially tert-butoxycarbonyl. The esterified carboxy is preferably esterified with alkyl having 1 to 4 carbon atoms, in particular methoxycarbonyl.

A &quot; particular compound of the invention &quot; is preferably in the free form. It is preferably in the unprotected form. is preferably hydroxymethyl optionally protected. R2 is preferably hydroxy. R 3 is preferably ethyl or allyl.

Another subgroup of particular compounds of the invention is that of the compounds of formula

och3

on what

Sis is methoxymethyl; hydroxymethyl optionally protected by tert-butyldimethylsilyl; benzoyloxymethyl; acyloxymethyl having 1 to 5 carbon atoms in the acyl part thereof optionally containing either a dimethylamino group which may be optionally quaternized or a carboxy group which may be optionally esterified by alkyl of 1 to 4 carbon atoms or an amino group which may be optionally protected by tert-butoxycarbonyl; or aminooxyalloxymethyl; or is methoxy, formyloxy or hydroxy optionally protected by tert-butyldimethylsilyl and there is a single bond between positions 23 and 24, or RgS is absent and there is a double bond between positions 23 and 24; and

R 2 is ethyl or allyl, in the free form and, when such forms exist, in salt form.

Particular compounds of the invention may be obtained by a process comprising the following steps: a) for the preparation of a compound of formula I wherein it is hydroxymethyl, or 5: ¾ is optionally protected hydroxy and there is a single bond between positions 23 and 24 or S2 is absent and there is a double bond between positions 23 and 24, and

R 2 is as defined above for formula I (i.e. a compound Ia),

a corresponding compound having a formyl group at the 32-position, i.e. a compound II of formula II

Wherein R 2 'is optionally hydroxy protected and there is a single bond between positions 23 and 24, or is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above for formula I' b) for the preparation of a compound of formula I and R4. is hydroxymethyl protected, or R 2 is hydroxy optionally protected and there is a single bond between positions 23 and 24, or R 12 is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above under formula I , (ie a compound Ib) a protecting group is suitably introduced into a compound of formula Ia; 11 -

V

c) for the preparation of a compound of formula I wherein R1 is optionally protected hydroxymethyl or formyloxymethyl, or R2 is optionally protected formyloxy or hydroxy and there is a single bond between positions 23 and 24, or R2 is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above under formula I, with the proviso that at least one of Rd and R 2 is formyloxymethyl or, respectively, formyloxy, (ie a compound Ic), a corresponding compound of formula I wherein R3 is optionally protected hydroxymethyl or formyloxymethyl or R4 is optionally protected formyloxy or hydroxy and there is a single bond between positions 23 and 24 or R8 is absent and there is a double bond between positions 23 and 24, and

R 1 is as defined above under formula I, with the proviso that at least one of R 1 and R 2 is other than formyloxymethyl or, respectively, formyloxy (i.e., compound Id); d) for the preparation of a compound of formula I in which R? is amino-oxyalkyloxymethyl, or R3 is optionally protected hydroxy and there is a single bond between positions 23 and 24, or Rz is absent and there is a double bond between positions 23 and 24, and R3 is as defined above under formula I (ie a compound Ie), is treated with an appropriate oxalyl derivative and then with ammonia, a corresponding Ia compound; e) for the preparation of a compound of formula I, wherein Ra is methoxymethyl or optionally protected hydroxymethyl, or Re is methoxy or optionally protected hydroxy and there is a single bond between positions 23 and 24 or R 1 is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above under formula I, with the proviso that at least one of R 11 is methoxymethyl or methoxy (ie an If compound), methyl suitably a corresponding compound of 12

formula I wherein Rd is optionally protected hydroxymethyl or is optionally protected hydroxy and there is a single bond between positions 23 and 24, or Ra is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above under formula I, (ie an Ig compound);

f) for the preparation of a compound of formula I wherein Si is acyloxymethyl wherein the acyl moiety optionally contains either a dimethylamino group which may be optionally quaternized, or a carboxy group which may be optionally esterified, or one or more amino groups and or optionally protected hydroxy, or R3 is optionally protected hydroxy and there is a single bond between positions 23 and 24 or R3 is absent and there is a double bond between positions 23 and 24, and R3 is as defined above under formula I (ie a compound of formula Ih), a corresponding compound of formula Ia is suitably acylated, and if desired, a compound of formula Ih wherein Râ, is as defined above is quaternized. contains a dimethylamino group;

and when a resulting compound of formula I has a protected hydroxy or hydroxymethyl group and / or a protected amino group, the protecting group (s) is optionally removed to provide a corresponding compound of formula I having one or more hydroxy or hydroxymethyl and / or an unprotected amino group (s) (ie a compound (1)); and the resulting compound of formula I is recovered in free form or, when such forms exist, in salt form. The invention further includes the use of a compound of Formula II as an intermediate, e.g. in the preparation of a compound of the invention as defined above, especially a particular compound of the invention as defined above.

A compound of formula II [which is a derivative of 12- (2'-cyclopentylvinyl) -1,2-dioxa-4-azatricyclo [22.3.1.0.02] octacos-18-ene or -14,18-diene , according to numeral in EP 184162] can be obtained by treating a compound of formula III

I am

NC-0 H

OCH3 where

R 1 is hydroxy or methoxy, R 2 ', or is optionally protected hydroxy and there is a single bond between positions 23 and 24, or is absent and there is a double bond between positions 23 and 24, and R 3 is as defined above under the formula I, [which is a 12- (2'-cyclohexylvinyl) -11,28-dioxa-4-azatricyclo [22.3.1.0 -octacos-18-ene or -14,18-diene derivative of according to the number in EP 1841623 with an acid having a non-nucleophilic anion.

This reaction constitutes a ring contraction. It is effected, e.g. as described above. Ο Ί //// .0 ο

Process variants a) to f) and deprotection will be seen as illustrations of appropriate derivatization of the cyclopentyl compound obtained by contraction of the ring.

A compound of formula III may be obtained, e.g. as described in EP 427660, by treating a corresponding 33-hydroxy compound - or with cyanogen bromide in the presence of a base; - or with thiophosgene, by reacting the resulting product with an inorganic azide and leaving the resulting unstable intermediate having an N-N-α-o-

II I

N-S at position 33 to decompose into a corresponding compound of formula III,

Where their preparation is not specifically described herein, eg in the Examples, the compounds used as starting materials are known or can be obtained in conventional manner from known compounds, eg starting from appropriate Streptomyces strains such as Streptomyces Tsukubaensis No. 9993 described, eg in Fujisawa EP 184162. Samples of

Fermentation Research Institute, Tsukuba, Ibaraki 305, Japan, according to the regulations of the Budapest Treaty under the NQ FERM BP-927 warehouse. This strain was redeposited on April 27, 1989, at the Agricultural Research Culture Collection International Depository, Peoria, Illinois, 61604, USA, under the Budapest Treaty under deposit WRLRRL 18488.

Process variants leading to &quot; particular compounds of the invention &quot; can be carried out in conventional manner. Process variant a) is a reduction in alcohol. It is preferably carried out in an inert solvent, e.g. a cyclic ether. The reducing agent is, eg, lithium-B-isopinocamphenyl-2-borabicyclo [3.3.1] nonylhydride (R-Alpinhidride®, Aldrich), in a cyclic ether, such as tetrahydrofuran or a borane-tert -butyl butylamine in a mixture of water and tetrahydrofuran. The temperature is preferably low, eg in the range of about -100 ° C to about -20 ° C, with the R-Alpinhydride® preferably between about -78 ° C, or between about -50 ° C to about -20 ° C, with the borane-tert-butoxycarbonyl-

butylamine, preferably at about 0 ° C. The reduction may alternatively be effected with diphenylsilane using a catalyst, such as a Wilkinson catalyst, preferably in an inert solvent such as an aromatic hydrocarbon, e.g. benzene. Diastereoisomeric mixtures (position 32) may be obtained, which may, if desired, be fractionated in conventional manner, e.g. by chromatography.

Process variant b) is an introduction of a protecting group. It may also be carried out in a conventional manner according to similar lines. Thus, for reactions involving a hydroxy group, in particular a hydroxy group at the 24-position and the substituent at the 32-position, the selective protection of only one of the two free hydroxy groups or the selective deprotection of only one of the two protected hydroxy groups, may be effected such that such substitution occurs only in the desired position. The reaction may, however, also be carried out with both unprotected positions. Mixtures of end products may thus be obtained: said mixtures may be separated in a conventional manner, for example by chromatography. The resulting final products further containing protecting groups may be subsequently deprotected, if desired. The reaction conditions may alternatively be selected so that they are removed simultaneously with or immediately after the reaction of the protecting groups (a process in a single reaction vessel). Depending on the reaction conditions used, such as the reaction duration, products are obtained which are predominantly mono-protected at the 32-position or diprotected at the 24-position and at the 32-position substituent. Process variant c) is a filler . It is carried out, for example, with a mixture of oxalyl halide, for example, chloride, and dimethylformamide (this mixture in solution in, for example, acetonitrile gives a Vilsmeyer complex). The reaction is carried out at ambient temperature or at a slightly reduced temperature, for example at a temperature from about room temperature to about 5 ° C. The formylation can also be carried out with the mixed anhydride of acetic acid and formic acid. A protective group may be removed at the same time. Process variant d) is an acylation. It is preferably carried out in an inert solvent, such as acetonitrile. The temperature is preferably about room temperature or slightly reduced, for example, between about 25Â ° and about 0Â °. The oxalyl derivative is preferably an oxalyl halide, for example, oxalyl chloride. After completion of the reaction, the mixture is stirred with ammonia. Thus a double bond may be formed between positions 23 and 24. Process variant e) is a methylation. It is preferably carried out in an inert solvent, such as a chlorinated hydrocarbon, for example dichloromethane. The methylating agent is preferably diazomethane in the presence of a Lewis acid, for example boron trifluoride-ether. The temperature is preferably from about 0 ° C to about room temperature. The protection group can be removed at the same time. Process variant f) is again an acylation. It is preferably carried out in an inert solvent, such as acetonitrile. The acylating agent is preferably an activated acyl derivative, such as an acyl halide or an anhydride. An acid scavenger such as dimethylaminopyridine or pyridine is usually employed. In addition, a carboxylic acid, such as glycine, protected on the amino moiety may be used, for example, tert-butoxycarbonyl or a carbodiimide, such as N-ethyl-N '- (3-dimethylaminopropyl) -carbodiimide or N, N'-dicyclohexylcarbodiimide, when necessary, in the presence of a base such as 4-dimethylaminopyridine, preferably in an inert solvent such as acetonitrile or a chlorinated hydrocarbon. The amino protecting group may be subsequently removed together with any hydroxy protecting group which may be present. If in the starting compound 1a is hydroxy and there is a single bond between positions 23 and 24, acylation may lead to the elimination of a water molecule at the 23,24 position, whereby a final compound can be formed in which Rz is absent and there is a double bond between positions 23 and 24. The optional subsequent guatemination step is also carried out in a conventional manner, eg by treatment with an alkyl iodide, preferably methyl iodide, in an inert solvent such as acetone. The opGional deprotection step may also be carried out in conventional manner. For the removal of, for example, tert-butyldimethylsilyl or tert-butoxycarbonyl, may be effected by treatment with hydrofluoric acid in a solvent such as acetonitrile. Depending on the 17

chosen reaction conditions (e.g., duration or temperature), the removal may be directed so that either all or only a few protecting groups are removed. Partial deprotection is particularly indicated when reacting a definitive hydroxy group in a subsequent reaction.

The following Examples illustrate the invention. They are not limiting. All temperatures are in degrees Celsius. In Nuclear Magnetic Resonance (NMR) spectra, all values of the chemical shifts are in ppm; the samples are measured in CDC13. the following abbreviations are used: BOC: cfr: db: OMe (or MeO): OtBDMS: Cf Ή FK'506: tert-butoxycarbonyl; colorless foamy resin, · double bond; methoxy; tert-butyldimethylsilyloxy; single bond, compound of formula

och3 18

(R) -hydroxy-12- [2 '- (4' '- (R) -hydroxy-3' '- (R) -methoxycyclohex- ) -1'-methyl-trans-vinyl] -23aÎ ±, β-dimethoxy-13β, 19,21β, 20-tetramethyl-11,28-dioxa-4-asatricyclo [22.3. (1)] octacos-18-trans-ene-2,3,10,16-tetraone (according to the numbering of atoms in EP 184162; however, numbering of the atoms of formula I) is used throughout the Examples; FR 520: as FK 506, but with CH 2 CH 2 (ethyl) instead of allyl at the 21-position in the formula cyclopentyl FK 506: compound of formula

cyclopent-1-FS520: as cyclopent-FK506, but with. . CH (ethyl) instead of allyl at the 21-position in the formula. 19

Example JL: 32-hydroxymethyl-cyclopentyl-FR520 [Formula I: R-hydroxymethyl; R2-hydroxy, single bond between positions 23 and 24; R 3 = ethyl] [Process variant a), reduction with a Wiperkinson catalyst].

A stirred solution of 25 g of 32-formyl-cyclopentyl-FR 520 (compound of formula II, Example 12 in EP 427 680) in 150 ml of benzene is cooled to 15 ° and treated successively with 7.25 ml of diphenylsilane and then with 200 mg of tris (phenylphosphine) rhodium (I) chloride and the cooling bath is removed immediately below. The resulting exothermic reaction occurs with increasing gas formation and the temperature rises to 29ø. 40 minutes after addition the reaction mixture is extensively concentrated to a bath temperature of not more than 40ø and a pressure of 30-40 mbar. The residue is taken up in 150 ml of acetonitrile, 5 ml of a 40% (w / w) aqueous solution of hydrofluoric acid are added in 5 minutes, and the mixture is stirred for another 25 minutes. The reaction mixture is poured into a well stirred mixture of 850 ml of ethyl acetate and 500 ml of saturated aqueous sodium bicarbonate solution and the resulting solution is thoroughly mixed. The aqueous phase is separated and extracted with 100 ml of ethyl acetate. The combined organic phases are successively washed with 100 ml of a saturated aqueous solution of sodium chloride and twice with portions of 100 ml of a saturated aqueous solution of sodium chloride, dried over sodium sulfate, filtered and evaporated The solvent was removed under reduced pressure. The residue is subjected to column chromatography (eluent: 1: 1 ethyl acetate / hexane) and the title compound is obtained as a diastereoisomeric mixture (cfr):

Diastereoisomer A: NMR (approximately 2: 1 mixture of conformers): main conformer: 4,258 (d, J = 1 Hz, 10-OH); minor conformer: 4,813 (d, J = 1.5H, 10-OH); C43 NMR (approximately 2: 1 mixture of conformers): main conformer: 213.4 (C-22); 196.2 (C-9); 159.05 (C-1); 164.8 (C-8); 97.1 (C-10)? 20 vj

Diastereoisomer B:% NMR (mixture approximately 2: 1 conformers): major conformer: 4,297 (d, J = 1 Hz, 10-OH); minor conformer: 4,795 (d, J = 1 Hz, 10-OH);  € ƒâ € ƒâ € ƒNMR (approximately 2: 1 mixture of conformers): main conformer: 213.4 (C-22) 196.12 (0-9)? 169.05 (C-1); 164.74 (C-8); 97.13 (C-10). The starting material is obtained in the following manner: (a) A solution of 2 g of 24-O-tert-butyldimethylsilyl-FR-20 and 0.94 g of 4-dimethylaminopyridine in 100 ml of dichloromethane was treated with a solution of 0.4 g of cyanogen bromide in 15 ml of dichloromethane and the mixture was stirred at room temperature for 20 minutes. The mixture is filtered on silica gel (eluent: n-hexane / ethyl acetate of acetic acid) and the solvent is removed from the relevant fraction under reduced pressure. 24-O-tert-butyldimethylsilyl-33-O-cyano-FS520 is obtained as a colorless foamy resin: α-NMR: mixture of rotamers: 4.3 (m; H-33); b) 0.5 g of the compound obtained in (a) above is dissolved in a mixture of 50 ml of acetonitrile and 2 ml of 40 wt% aqueous hydrofluoric acid and the mixture is stirred for 2.5 hours at room temperature . The reaction mixture is then partitioned between ethyl acetate and an aqueous saturated sodium bicarbonate solution, the aqueous phase is discarded and the organic phase is washed with saturated aqueous sodium chloride solution, dried dried over sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure. 32-formylcyclopentyl-FR 520 is obtained as a colorless foamy resin from the residue by silica gel column chromatography (eluent: n-hexane / ethyl acetate): NMR of the mixture of conformers : 9.64 (d, J = 2H, CHO); 2.87 (m, H-32); 2.67 (m, H-30).

Example 2: 24-O-fcb-butyldimethylsilyl-32-hydroxyformyl-cyclopentyl-FA520 [Formula I: R1 = hydroxymethyl; Sz = OtBDMS, single bond between positions 23 and 24, R3 = ethyl] [Process variant a), reduction with R-Alpinhidride® or a borane-tert-butylamine complex].

A solution of 180 mg of 24-O-tert-butyldimethylsilyl-32-formyl-cyclopentyl-FR52 (compound of formula II: Example 71 in EP 427680) in 15 ml of tetrahydrofuran at -78 ° 0.5 ml of a 0.5 M solution of 1-thio-B-isopinocanphenyl-2-borabicyclo [3.3.1] nonylhydride (R-Alpinhydride®, Aldrich) in tetrahydrofuran and the mixture is stirred for 1 hour , at this temperature. The reaction mixture is partitioned between 1 N HCl and ethyl acetate, the organic phase is separated, washed with 1 N HCl and a saturated aqueous solution of sodium chloride is dried over sodium sulfate, filtered, under reduced pressure.

The residue was chromatographed on silica gel column chromatography (eluents n-hexane / ethyl acetate 1: 5: 1). The title compound is obtained (cfr).

In one embodiment the reaction is carried out in 40 ml of tetrahydrofuran and 10 ml of water at a temperature ranging from 0-5Â ° C with 1 molar equivalent of a borane-tert-butylamine complex. Stirring is continued for 5 minutes at 0-5 °, the mixture is added to a mixture of 100 ml of 1M hydrochloric acid and 150 ml of ethyl acetate and stirred for 15 minutes, the organic phase is obtained and the title compound is obtained as follows: 1 H-NMR: δ 5.42 (d, J = 9 Hz, H-29); 5.21 (d, J = 7.5 Hz, H-26); 4.81 (d, J = 10 Hz, H-20); 3.82 (dxd, J = 9 and 1 Hz, H-14); 3.51 (m, CH 2 OH). The starting material is obtained as follows: A solution of 0.5 g of 32-formyl-cyclopentyl-FR 520 [compound obtained in (b) in Example 1] is stirred for 15 hours at room temperature , Tert-butyl dimethylsilyl chloride (5 g) and imidazole (0.25 g) in dry dimethylformamide (20 ml), and then partitioned between water and 22

ethyl acetate. The organic phase is separated, washed with an aqueous solution, saturated sodium chloride, dried over sodium sulfate, filtered and the filtrate is concentrated under reduced pressure. 24-O-tert-butyldimethylsilyl-32-cyclopentyl-FR52 is obtained from the residue as a colorless foamy resin followed by silica gel column chromatography (eluent: n-hexane / ethyl acetate 2 : 1): NMR of rotamer blends: 9.65 (d, J = 2 Hz, CHQ); 5.3S (d, J = 9 Hz, H-29); 5.01 (d, J = 7.5 Hz, H-26); 4.81 (d, J = 10 Hz, H-20); 3.82 (dxd, J = 9Hz, H-14).

Example 3; 32-tert-butyldimethylsilyloxymethyl-cyclopentyl-FR 520 [Formula I: R 4 = -CH 2 O-DBMS; Rz = hydroxy, single bond between positions 23 and 24; R 1 = ethyl] [process variant b), protection].

A solution of 0.5 g of 32-hydroxymethylcyclopentyl FR520 (compound of Example 1) in 30 ml of dimethylformamide is treated with 3 molar equivalents of tert-butyldimethylsilyl chloride and 3 molar equivalents of imidazole and the mixture is stirred for 2.5 hours at room temperature. The resulting mixture is partitioned between ethyl acetate and water, the organic phase is decanted dried over sodium sulfate, filtered and concentrated. The title compound (cf) is obtained from the residue by silica gel chromatography (eluent * n-hexane / ethyl acetate 3: 1). 1 H-NMR

Diaestereoisomer A: 5.34 (d, J = 2.5 Hz, H-26); 4.6 (db, J = 4 Hz, H-2)? 4.44 (db, J = 13 Hz, H-6 eau.); 4.25 (bs, 10-OH); 3.92 (m, H-24); 3.69 (d, J = 9 Hz, H-14); 3.59 (dxd, J = 2.5 and 10 Hz, H-15); 3,465 (d, J = 6 Hz, -CH2 OSi); 3.40 and 3.31 (s and s, 2x-OCH3); 0.9 (s, t-butyl); 0.04 (s, Si-CH 3). I 23

Diastereoisomer B: 5.34 (d, J = 2.5 Hz, H-26); 4.6 (db, J = 4 Hz, H-2); 4.44 (db, J = 13 Hz, H-6 equ.); 4.20 (bs, 10-OH); 3.92 (m, H-24); 3.69 (d, J = 9 Hz, H-14); 3.59 (dxd, J = 2.5 Hz, H-15); 3,465 (d, J = 6 Hz, -CHtOSi); 3.40 and 3.31 (s and s, 2x-OCH3); 0.9 (s, t-butyl); 0.04 (s, Si-CH *).

Example 4: 24-O-formyl-32-formyloxymethyl-cyclopentyl-FR520 [Formula I: R4 formyloxymethyl; S3 = formyloxy, single bond between positions 23 and 24; R 3 = ethyl 3 [process variant c), formylation]

A solution of 0.5 g of 4-dimethylaminopyridine (0.5 g) and formic acid / acetic acid (0.3 g) was treated with 0.5 g of 32-hydroxymethylcyclopentyl-FR520 (compound of example 1) in 30 ml of acetonitrile mixed and stirred at room temperature for 30 minutes. The mixture is partitioned between ethyl acetate and 1N hydrochloric acid, the organic phase is decanted, washed with brine, filtered and concentrated under reduced pressure. The title compound (cf) is obtained from the residue by silica gel column chromatography (eluent: n-hexane / ethyl acetate 2: 1): NMR of a mixture of conformers; 8.08 and 8.01 (s and s, 2X-O-CHO); 4.90 (d, J = 9 Hz, H-20); 4.55 (m, H-2); 4.44 (db, J = 13Hs, H-6 equ.); 4.05 (d, J = 7.5Hz, -CHa-OCO-); 3.77 (dxd, J = 9 and 1 Hz, H-14).

Example 5: a) 32-aminooxalyloxymethyl-24-dehydroxy- Δ3 -cyclopentyl-FR520 and b) 32-aminooxyalloxymethylcyclopentyl-FR520 [Formula I: R1 = aminooxyalloxymethyl; Rz = absent,

double bond between positions 23 and 24 and, in each case, R1 = hydroxy, single bond between positions 23 and 24; R 3 = ethyl [process variant d) treatment with the oxalyl derivative and then ammonia].

0.5 g of 24-0-tert-butyldimethylsilyl-32-tert-butyldimethylsilyloxymethylcyclopentyl-FR 520 (compound of example 16) in 30 ml of acetonitrile is treated with 0.2 ml of oxalyl chloride, and the mixture was stirred for 3 hours at room temperature. The reaction mixture is then added to a vigorously stirred mixture of 200 ml of ethyl acetate, 20 ml of an aqueous saturated ammonia solution and 100 ml of a saturated aqueous solution of sodium chloride, and the solution for 20 minutes. The components are not dissolved, the organic phase is decanted, washed successively with water, 1N hydrochloric acid and water, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue is taken up in 30 ml of acetonitrile, treated with 1.5 ml of 40% w / w aqueous hydrofluoric acid and stir the solution for 3 hours at room temperature. The solution is partitioned between ethyl acetate and saturated aqueous sodium bicarbonate solution, the organic phase is decanted, dried over sodium sulfate, filtered and concentrated. The title compounds (cfr) are obtained from the residue by silica gel column chromatography (eluent: n-hexane / ethyl acetate 2: 1): 1 H NMR:

Compound a): mixture of conformers: 6.97 and 5.77 (b, -COWHs); 6.83 (dxd, J = 6 and 15 Hz, H-24); 6.18 (dxd, J = 1 and 15 Hz, H-23); 4.20 (m, -CH2 C = O);

Compound b): mixture of conformers: 6.97 and 5.92 (b, -CONH), 4.60 (d, J = 3 Hz, H-2), 4.44 (db, J = 13 Hz, H-6 equ.), 4.19 (m, CH-zC = 0), 3.92 (m, H-24), 3.68 (d, J = 9Hz, H-14) τ 3.40 and 3.32 (2x s, 2x-OCH 3).

Example 6; 24-methoxy-32-methoxymethyl-cyclopentyl] -FR520 [Formula I: R4 = methoxymethyl; R 2 -methoxy, single bond between positions 23 and 24; R 3 = ethyl] [process variant e), methylation].

A solution of 1 g of 32-hydroxymethyl-cyclopentyl-FR-520 (compound of example 1) and 0.04 ml of trifluoro-trifluoride etherate in 50 ml of dichloromethane is cooled to 0-5 °, and treated is added with portions of a 10 ml (approximately 0.5 M) solution of diazomethane in dichloromethane, so that the initial yellow coloring which forms with the addition persists for as little time as possible. The mixture is then partitioned between ethyl acetate and a saturated aqueous solution of sodium bicarbonate, the organic phase is decanted, dried over sodium sulfate, filtered and concentrated. The title compound (cfr) is obtained from the residue by column chromatography on silica gel (eluent: n-hexane / ethyl acetate 2: 1): 1 H NMR: d, J = 8.8Hz, H-29); 5.18 (d, J = 7.6 Hz, H-26); 4.77 (d, J = 10.2 Hz, H-20); 3.81 (dxd, J = 9.7 / 1Hz, H-14); 2.67 (dxd, J = 13.5 / 8.4Hz, H-23).

Example 7: 24-O-tert-butyldimethylsilyl-32-isobutanoyloxy-methyl-cyclopentyl-FR520 [Formula I: R1 = isobutanoyloxymethyl; Rt = OtBDMS, single bond between positions 23 and 24; R 3 = ethyl] [process variant f), acylation].

0.5 g of 24-O-tert-butyldimethylsilyl-32-hydroxymethyl-cyclopentyl-FR 520 (compound of Example 2) and 0.5 g of dimethylaminopyridine in 30 ml of acetonitrile are treated with 0.3 ml of anhydride of isobutyric acid and the mixture is stirred for 4 hours at room temperature. The mixture is then partitioned between ethyl acetate and 1N aqueous hydrochloric acid, the organic phase is decanted, washed with 26β / 7 /

IK * &lt; A saturated aqueous solution of sodium chloride, filtered, and concentrated under reduced pressure. The title compound (cfr) is obtained from the residue by silica gel column chromatography (eluent: n-hexane / ethyl acetate 3: 1): 4H NMR: mixture of conformers: 3.95 (d, J = = 7Hz, -CHâ,ƒOCO-); 2.55 [sept, J = 7Hz, H-C (CH3) e3; 1.17 (d, J = 7 Hz, 2x-CH 3).

Example 8: 24-0-tert-butyldimethylsilyl-32 - [(M, M-dimethylamino) methylcarbonyloxymethylcyclopentyl] -FR520 [Formula I: R * = [(Î ±, Î'-dimethylamino) methyl] carbonyloxymethyl ; R2 = OtBDMS, single bond between positions 23 and 24; R 3 = ethyl] [process variant f), acylation.

A solution of 1 g of 24-O-tert-butyldimethylsilyl-32-hydroxymethylcyclopentyl-FR520 (compound of Example 2) in 35 ml of absolute pyridine with 1.2 equivalents of N, N'-dicyclohexylcarbodiimide , 1.2 equivalents of N, N-dimethylglycine and 0.2 equivalents of p-toluenesulfonic acid monohydrate and the mixture is stirred for 15 hours at room temperature. The reaction mixture is then partitioned between water and ethyl acetate, the organic phase is decanted, washed successively with 1N hydrochloric acid solution and saturated aqueous sodium chloride solution, dried over sodium sulfate sodium, filtered and concentrated under reduced pressure. The title compound (cf) is obtained from the residue by silica gel column chromatography (eluent: ethyl acetate / methanol 20: 1):

Characterization data: Deprotection gives the compound of Example 20.

Example 9: 32- [N-tert-butoxycarbonylamino) methyl] carbonyloxymethyl-24-tert-butyldimethylsilyl-cyclopentyl] -FR520 [Formula I: ΕΑ = ΒΟΟ-ΝΗΟΗ 2ΟΟΟΟΗε -; Rz = OtBDMS, link 27

between positions 23 and 24? R 3 = ethyl] [process variant f), acylation]

A solution of 0.5 g of 24-O-tert-butyldimethylsilyl-32-hydroxymethyl-4-cyclopentyl-FR 520 (compound of Example 2), 0.3 g of N- -BOC-glycine and 0.5 g of dicyclohexylcarbodiimide in 30 ml of acetonitrile. Then, the mixture is diluted with 30 ml of ether and the insoluble compounds are filtered. The solution is then partitioned between ethyl acetate and 1N aqueous hydrochloric acid, the organic phase is decanted, washed with a saturated aqueous solution of sodium chloride, filtered, dried over sodium sulfate and concentrated under reduced pressure. The title compound (cfr) is obtained from the residue by silica gel column chromatography (eluent: n-hexane / ethyl acetate 3: 1): ¹H NMR: mixture of conformers: 4.81 ( d, J = 10Hz, H-20), 4.44 (H-2); 4.42 (H-6 egu.); 4.03 (d, J = 7Hs, -CH20 C = O); 3.91 (d, J = 6H2, -NH-CH2 C = O); 3.82 (dxd, J = 1.5 and 9 Hz, H-14); 3.40 and 3.32 (s and s, 2x-OCH3); 1.46 (s, t-butyl-O-C = O); 0.87 (s, t-butyl-Si).

Example 10: 32- (Ν, Μ, Ν-trimethylammoniomethyl) carbonyloxymethylcyclopentyl-FR 520 [Formulai: R * = (H, C) 3 N + CH (CH 2) CH 2 -; R2 = OH, single bond between positions 23 and 24; R3 = ethyl; in the form of quaternary iodide ammonium salt] [process variant f), optional quaternization].

A solution of 0.5 g of 32- (Î ±, Î ± -dimethylaminomethyl) carbonyloxymethylcyclopentyl FR520 (compound of Example 20) in 25 ml of acetone and 10 ml of diethyl ether is treated at with 0.5 ml of methyl iodide and stirring solution for 8 hours. Thereafter, the reaction mixture is diluted with several portions of diethyl ether, 28

the precipitate is collected by suctioning, washed repeatedly with diethyl ether and dried under high vacuum. The title compound (amorphous pale yellow powder) is obtained: δ-NMR: mixture of conformers: 3.66 (S, N (CH 3) 3 +); 4.99 (bs, N-CH * -CO)

Example 11: 32-Isobutanoyloxymethylcyclopentyl-FR-52

[Formula I: R isobutanoyloxymethyl; Rz = OH, single bond between positions 23 and 24; R 3 = ethyl] [process variant: deprotection.

A solution of 0.8 g of 24-O-tert-butyldimethylsilyl-32-isobutanoyloxymethylcyclopentyl-FR520 (compound of Example 7) in 40 ml of acetonitrile is treated with 2.5 ml of a 40% solution of % w / w of hydrofluoric acid, and the mixture is stirred for 6 hours at room temperature. The reaction mixture is then partitioned between ethyl acetate and saturated aqueous sodium bicarbonate solution, the organic phase is decanted, dried over sodium sulfate, filtered and concentrated. The title compound (ofr) is obtained by silica gel column chromatography (eluent: n-hexane / ethyl acetate 2: 1): 1 H NMR: The **

* ******** · - * -H * zzzzzzzz s- x c. 2 cu x cu * * *

The following compounds of Formula I are obtained analogously:

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Example 19: 5.41 (d, J = 9 Hz, H-29); 4.82 (d, J = 10 Hz, H-20); 4.44 (H-2); 4.41 (H-6 equ.); 4.04 (H-24); 3.95 (d, J = 7 Hz, -CHOOC = O); 3.82 (dxd, J = 1.5 and 9 Hz, H-14); 3.40 and II o oo

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The compounds of the invention possess pharmacological activity. They are indicated for use as pharmaceuticals.

In particular, they possess immunosuppressive, anti-Droliferative and anti-inflammatory activity. Anti-inflammatory activity may, for example, be determined in the following test methods: 1. Oxazolone (mouse) allergic contact dermatitis [The test method is as described in FM Dietrich and R. Hess, Int. Arch . AIlerαν, 38 (1970) 246-259]:

The compounds deduce (reveal) in this assay an activity (inhibition of inflammatory swelling) of between about 20% and about 63%, with a single topical application in the form of a 0.01% solution. 2. Allergic contact dermatitis caused by DHFB (domestic pig): (The test method is as described in, for example, EF 315978):

Two topical applications of a 0.4% formulation of the compounds have resulted in the inhibition of the inflammatory reaction from about 16% to about 52%. 3. Irritant contact dermatitis induced by inhibition of forboi ester (TPA) (mouse) (The assay method is as described in e.g. EP 315978):

The compounds deduce (reveal) in an assay, with a single application of a 3.6% formulation, an inhibition of the inflammatory reaction, from about 10% to about 30%. Immunosuppressive and antiproliferative activity may, e.g. be determined in the following test methods:

(I.e.

1. Oroliferative response of 1infocytes to the stimulation of alloenics in the reaction with mixed lymphocyte (MLR) in vitro [T. Meo. &quot; The MLR in the Mouse &quot;, Immunoloaical Methools, L. LefKovits and B. Pernis, Eds., Academic Press, N.Y. (1979), 227-239]:

The compounds deduce (reveal) in this assay the suppression of 1 mixed infocytes [01] at a dosage of about <0.0008 to 1 g / ml to about 0.09 Âμg / ml. 2. Inhibition of primary human immune response in sheep erythrocytes in vitro

The assay method is as described in R.I. Mishell and R.W. Duton »Science 153 (1966) 1004-1006; R.I.

Mi Shell and R.W. Duton. Exp. Med. 126 (1967) 423-4423;

The compounds are active in this assay with an IC 50 of about 0.0024 μg / ml and about 0.32 μg / ml. 3. Inhibition of proliferation of human keratinocytes (the assay method is as described in e.g. EP 315978)

The compounds are active in this assay in concentrations ranging from about 1 μg / ml to about 10 μg / ml, resulting in an inhibition of about 30% to about 90%. The compound of Examples 1 and 12 (32-hydroxymethylcyclopentyl-FR 520) is the preferred compound as an immunosuppressive agent. For example, it has been determined that in the DWFB (domestic pig) allergic contact dermatitis test, this compound as a 0.4% preparation has an activity better than a preparation corresponding to 1.2% dexamethasone. It is therefore indicated that for this indication, the compound of Examples 1 and 12 may be administered to larger mammals, for example, to man, by similar administration forms, in similar or lower dosages than those conventionally employed with dexamethasone.

The compounds of the present invention in free form or where such forms exist in the form of pharmaceutically acceptable salts are therefore indicated as immunosuppressive and antiproliferative agents and as anti-inflammatory agents for use in the prevention and treatment of conditions requiring immunosuppressive and inflammatory conditions, such as: a) prevention and treatment of: - resistance in transplantation of organs or tissues, eg heart, kidneys, liver, bone marrow and skin; disease of patient rejection of grafts, such as the following to bone marrow grafts; - autoimmune diseases such as rheumatoid arthritis, systemic lupus erythematosus, Hashimoto's thyroidism, multiple sclerosis, severe myasthenia, type I diabetes and uveitis; - skin manifestations of immunologically mediated disorders; b) treatment of inflammatory and hyperproliferative diseases of the skin, such as psoriasis, atopic dermatitis, contact dermatitis, and other eczemas-dermatoses, seborrheic dermatitis, Lichen planus, pemphigus, puffy pemphigoid inflammations, epidermolysis bullosa, urticaria, angiodemas, vasculitides, erythema, cutaneous eosinophils, lupus erythematosus and acne; c) Alopecia areata.

The compounds may be administered systemically or topically. For the above indications, the appropriate dosage will of course vary, depending, for example, on the host, the mode of administration and the nature and severity of the condition being treated. However, in general satisfactory results are obtained systemically at daily dosages ranging from about 0.15 mg / kg to about 1.5 mg / kg of body weight of the animal. A daily dosage indicated in larger mammals is in the range of about 0.01 mg to about 100 mg, conveniently administered, for example, in divided doses up to four times daily or in the delayed form. For topical use satisfactory results are obtained with local administration with a concentration of 1 to 3% of active substance several times a day, for example 2 to 5 times a day. Examples of indicated dosage forms are lotions, gels and creams.

The compounds of the invention may be administered by any conventional route, in particular, enterally, for example, orally, for example in the form of tablets or capsules, or topically, e.g.

form of lotions »gels or creams.

The pharmaceutical compositions, for example, for topical application, comprising a compound of the invention in free form and, when such forms exist, in pharmaceutically acceptable salt form in association with at least one pharmaceutically acceptable carrier or diluent 'may be produced in a conventional manner, for example, by admixing the pharmaceutically acceptable diluent with a carrier. Dosage unit forms contain, for example, from about 0.0025 mg to about 50 mg of active ingredient. Topical administration is effected for example on the skin. Another form of topical administration is to the ocular globe, for the treatment of immunocompromised eye states, such as autoimmune diseases e.g., uveitis, keratoplasty and chronic keratitis; allergic conditions, for example vernal conjunctivitis; inflammatory conditions and corneal transplants through topical administration to the surface of the eye of a compound of the invention in a pharmaceutically acceptable ophthalmic vehicle. The ophthalmic vehicle is such that the compound is maintained in contact with the ocular surface for a period of time sufficient to allow the compound to penetrate the corneal and internal regions of the eye, for example, the anterior, posterior, vitreous, in the aqueous humor, in the vitreous, cornea, iris / ciliary, lenses, choroid / retina and sclera. The pharmaceutically acceptable ophthalmic vehicle may be, for example, an ointment, a vegetable oil, or an encapsulating material.

While anti-inflammatory, immunosuppressive and anti-proliferative activity is the main activity of the compounds of the invention, they also have some degree of activity in increasing the sensitivity to, or enhancement of efficacy, of chemotherapeutic drug therapy. This activity may, for example, be determined according to the test methods described in EP 360750.

The compounds of the invention are therefore indicated for use in reversing therapeutic drug resistance of various types, for example, acquired or cream, or in increasing sensitivity to drug therapy, for example, as a means of reducing levels out of 37

for example, in the case of anti-neoplastic and cytostatic drug therapy, as a means of decreasing the toxicity in general caused by the medicament, and more specifically, as a means of reversing or reducing resistance, including inherent resistance and · Acquired, to chemo. The invention thus also relates to the use of a compound of the invention as a pharmaceutical, such a compound for use as a pharmaceutical, to the use of such a compound for the preparation of a pharmaceutical composition, which comprises admixing with, at least , a pharmaceutically acceptable carrier or diluent, and to a process for the preparation of a pharmaceutical composition comprising mixing a compound of the invention together with at least one pharmaceutically acceptable carrier or diluent.

Claims (4)

IV REPORTS: Ia - A process for the preparation of tricyclic compounds containing heteroatoms, in particular, 12- (2'-cyclopentylvinyl) -11,28-dioxa-4-azatricyclo [22.3.1.04,4] -octacosene derivatives -18 or 12- (2'-cyclopentylvinyl) -11,28-dioxa-4 -9-azatricyclo [22.3.1.0 '] -octacosadiene-14,18 or 11- (2'-cyclo pentylvinyl) -10,27-dioxa-4 '-azatri-cyclo- [21.3.1.0'] heptacosene-17 or 11- (2'-cyclopentylvinyl) -10,27-dioxa-4-azatricyclo [21.3 .1.0 '] -heptacho-sadien-13-17, with the proviso that its cyclopentyl moiety is other than 3-formylcyclopentyl characterized in that the ring-shrinking reaction of an appropriate 12- ( 2-cyclohexylvinyl) -11,28-dioxa-4-azatricyclo [22.3.1.0 4 ']] octacosen-18, or 12- (2'-cyclohexylvinyl) -1,2dio-dioxa Azatricyclo [22.3.1.0 '] -octacosadiene-14,18, or a suitable derivative of 11- (2S-cyclohexylvinyl) -10,27-dioxa-4,4-azatricyclo [22.3.1.0 '] -heptaco 17-ene, or 11- (2'-cyclohexylvinyl) -10,27-dioxa-4-azatricyclo [22.3.1.0 '] hepta-cosadiene-13.17 and the compound of the resulting cyclopentyl. The A process according to claim 1, characterized in that one of the following compounds is obtained as a final product: a 17-alkyl (or 17-alkenyl) -1-hydroxy-14-oxy-12- (2 ' 1-methyl-trans-vinyl) -23,25-dimethoxy-13,19,21,27-tetrahydronaphth-11,28-dioxa-4-azatricyclo [22.3.1.0 ] -trans-octa-cosene-18-2,3,10,16-tetrone, or a 17-alkyl (or 17-alkenyl) -1-hydroxy-12- (2'-cyclopentyl-1'-methyl 3-yl) -23,25-dimethoxy-13,19,21,27-tetramethyl ester (22.3.1.0 ') -trans-octacosadiene-14,18 (or trans-18-octacosene) -2,3,10,16-tetrone, with the proviso that (17β-alkenyl) -lbeta (17β-alkenyl) -cyclopentyl-17β-allyl-cyclopentyl -hydroxy-14alpha-oxy-12- [2 '- (cyclopent-1' '- (R) -methyl-trans-vinyl] -23alpha, 25beta-dimethoxy-13-alpha 19,21alpha] -tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.0 (4 ')] transoctacosene-18-9alphaH, 24betaH-2,3,10,16-tetrone or a derivative of 17α-alkyl (or 17alpha-alkenyl) -betahydroxy-12- [2 '- (cyclopent-1' '(R) -yl) -1'-methyl-trans-vinyl] -23alpha, 25beta-dimethoxy -6-alpha9,21alpha, 27beta-tetramethyl-11,28-dioxa-4-azatryl-cyclo- [22.3.1.0 '] -translocazine-diene-17,18- (or-trans-18-octacosene) -9aÎ ± H, 24betaH -2,3,10,16-tetrone, with the proviso that its cyclopentyl moiety is other than 3-formylcyclopentyl moiety. 4. A process for the preparation of a compound of formula I in which it is methoxymethyl; optionally protected hydroxymethyl; acyloxymethyl wherein the acyl part optionally contains or a dimethylamino group which may be optionally quaternized, κ or a carboxy group which may be optionally esterified, or one or more amino and / or hydroxy groups which may be optionally protected; or amino-oxyalkyloxymethyl; or is optionally protected methoxy, formyloxy or hydroxy and there is a single bond between positions 23 and 24 or R2 is absent, and there is a double bond between positions 23 and 24; and R3 is methyl, ethyl, n-propyl or allyl; in free form and, when such forms exist, in salt form according to claim 1, characterized in that it comprises a) for the preparation of a compound of formula I wherein R 1 is hydroxymethyl, or R 2 is optionally protected hydroxy and there is a single bond between positions 23 and 24 or R2 is absent and there is a double bond between positions 23 and 24; and. Rg is as defined in this claim in formula I (i.e., a compound of formula Ia), if appropriately reducing a corresponding compound of formula II in which R 2 'or is optionally protected hydroxy and there is a single bond between positions 23 and 24, or is absent and there is a double bond between positions 23 and 24, and α 1 is as defined in this claim under formula I; b) for the preparation of a compound of formula I wherein R1 is protected hydroxymethyl, or R2 is hydroxy optionally protected and there is a single bond between positions 23 and 24, or R2 is absent and there is a double bond between positions 23 and 24, and R6 is as defined in this claim under formula I, if a protecting group is suitably introduced into a compound of formula Ia; c) for the preparation of a compound of formula I wherein R 1 is hydroxymethyl or formyloxymethyl optionally protected, or R 2 is formyloxy or optionally protected hydroxy and there is a single bond between positions 23 and 24, or R 2 is absent and there is a ' double bond between positions 23 and 24, and is as defined in this claim under formula I, with the proviso that at least one of R1 and R2 is formyloxymethyl or formyloxy, a corresponding compound of formula I wherein is optionally protected hydroxymethyl or formyloxymethyl or R 2 is formyloxy or optionally protected hydroxy and there is a single bond between positions 23 and 24, or R 2 is absent and there is a double bond between positions 23 and 24, and R 2 is as defined in this claim under formula I, with the proviso that at least one of R1 and R2 is other than formyloxymethyl or, respectively, Formyloxy; d) for the preparation of a compound of formula I wherein R1 is amino-oxyalloxymethyl, or R2 is optionally protected hydroxy and there is a single bond between positions 23 and 24, or R 2 is absent and there is a double single bond between positions 23 and 24, and R 3 is as defined in this claim under formula I, if a compound of the formula la is treated with a oxalyl and then with ammonia; e) for the preparation of a compound of formula I, wherein R 1 is methoxymethyl or optionally protected hydroxymethyl, or R 2 is methoxy or optionally protected hydroxy and there is a single bond between positions 23 and 24, or R 2 is absent and there is a double bond between positions 23 and 24, and R 3 is as defined in this claim under formula I, with the proviso that at least one of R 1 and R 2 is methoxymethyl or, respectively, methoxy, suitably methylating a compound corresponding compound of formula I wherein R 1 is optionally protected hydroxymethyl, or R 2 is hydroxy optionally protected and there is a single bond between positions 23 and 24, or R 2 is absent and there is a double bond between positions 23 and 24, and R 1 is as defined in this claim under formula I; f) for the preparation of a compound of formula I wherein R 1 'is acyloxymethyl wherein the acyl part optionally contains either a dimethylamino group which may be optionally quaternized or a carboxy group which may optionally be esterified or one or more amino and / or hydroxy optionally protected, or R2 is hydroxy optionally protected and there is a single bond between positions 23 and 24, or R2 is absent and there is a double bond between positions 23 and 24, and is as defined in this claim under formula I (i.e., a compound of formula Ih), a corresponding compound of formula Ia is suitably acylated and, if desired, quaternizing a compound of formula Ih wherein R- contains a dimethylamino group; and when a resulting compound of formula I has a protected hydroxy or hydroxymethyl group and / or a protected amino group optionally removing the protecting group (s) to give a corresponding compound of formula I having one or more hydroxy or hydroxymethyl groups and / or an unprotected amino group or groups? and recovering the resulting compound of formula I in the free form or, when such forms exist, in salt form. The 5. A process for the preparation of the compound of formula I according to claim 4, characterized in that 32-hydroxymethylcyclopentyl-FR 520 is obtained as the final product in the free form and, when such forms exist, the salt form. 6. A process for the preparation of pharmaceutical compositions having immunosuppressive and antiproliferative activity characterized in that a compound prepared according to claim 1 is mixed with at least one pharmaceutically acceptable carrier or diluent. 7. A process for the preparation of the cyclopentyl compound used as an intermediate for the preparation of the compounds of formula (I) according to claim 1, characterized in that a 12- (2'- cyclohexylvinyl) -1,28-dioxa-4-azatricyclo [22.3.1.0] octacosen-18 or a 12- (2-cyclohexylvinyl) -11,28-dioxa-4-azatricyclo [22.3. 1.04 '] - octacosa-diene-14,18-diene, or a derivative of 11- (2-O- cyclohexylvinyl) -10,27-dioxa-4-azatricyclo [22.3.1.oool] -heptacosene-17 or 11- (2'-cyclohexylvinyl) -10,27-dioxa-4- -48-azatricyclo [22.3.1.0 '] heptacosadiene-13.17. Lisbon, July 1, 1991 / The Official Agent of Industrial Property Américo da Silva Ca rvalh a Agssts Office of Propaganda for the Indus- trial Industry of the Border, No. IS? 2, at 1o0 LISBON Tel8.3877373 &gt; 38774SI