NOVEL MACROCYCLIC COMPOUNDS AND NOVEL METHOD OF TREATMENT

Abstract: Compounds of formula (I), wherein R<1> and R<2> independently represent H or OH, or a second carbon-carbon bond; R<3> represents optionally substituted methyl, ethyl or propyl; R<4> represents H; R<5> and R<6> represent a second carbon-carbon bond; R<7> represents H or OH; R<8> represents OCH3; R<9> represents OH or OCH3; X and Y independently represent O or (H, OH); n represents 1 or 2; and in addition some of the substituents form rings with each other; with various provisos; are indicated in the treatment of immunodepression. A number of novel compounds of formula (I) are also provided.

Description: NOVEL MACROCYCLIC COMPOUNDS AND NOVEL METHOD OF TREATMENT This invention relates to novel pharmaceutical uses o certain known macrocyclic compounds, and to nove macrocyclic compounds which have the same novel utility. 5 European patent application No 184162 (to Fujisaw Pharmaceuticals Co Ltd) discloses several macrocycli compounds which are derivatives of 12-(2-cyclohexyl- 1-methylvinyl)-13,19,21,27-tetramethyl-ll,28-dioxa-4- azatricyclo[22.3.l.0 4 ' 9 ]octacos-18-ene (numbere 10 FR-900506, FR-900520, FR-900523 and FR-900525) and whic are isolated from microorganisms belonging to the genu Streptomyces. The macrocyclic compounds and a number o their derivatives are indicated as immunosuppressiv agents. ,c International patent application No WO 89/05304 (als European Patent application No 323042, both to Fisons pic) discloses a large number of macrocyclic compounds which ma be derived from those disclosed in European patent application No 184162. Again, the compounds are primarily 2 Q indicated 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 patent 2 5 applications mentioned above may be used in the production of the novel compounds of the present invention. Alternatively, the novel compounds may be produced by total synthesis. We have now found a novel group of macrocyclic compounds which act as antagonists of immunosuppressive compounds, particularly macrocyclic immunosuppressive compounds including derivatives of 12-(2-cyclohexyl-l-methylvinyl)-13,19,21,27-tetramethyl- 11,28-dioxa-4-azatricyclo[22.3.1.0 4 > 9 ]octacos-18-ene and rapamycin, as shown in the mixed lymphocyte reaction (MLR) (described in WO 89/05304, Example A). The novel group of compounds are therefore useful inter alia in the treatment of immunodepression or a disorder involving immunodepressio . Thus, according to the present invention, there is provided the use of a compound of formula I, wherein R 1 and R 2 independently represent H or OH, or they may together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 3 represents methyl optionally substituted b -C0 2 H or an ester or amide thereof; ethyl optionally substituted by O, OH or -C0 2 H or an ester or amide thereof; propyl optionally substituted by OH or O; or allyl optionally substituted by OH; R 4 represents H; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H or OH; R 8 represents OCH 3 ; R 9 represents OH or OCH 3 ; X represents O or (H,OH) ; Y represents O or (H,OH) ; and n represents 1 or 2; in addition to their significances above R 1 and R 5 may together represent an oxygen atom, in which case R 6 and R 7 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 and R 8 may together represent an oxygen atom; ar R 3 , R 4 and Y, together with the carbon atoms to which they are attached, may represent a methyl-substituted furanyl ring; provided that i) when R 2 represents H; R 3 represents methyl, ethyl, propyl or allyl; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 8 represents OCH 3 ; and Y represents O; then R 7 represents OH; and ii) when n is 1, then R 3 is not methyl or ethyl; in the manufacture of a medicament for the treatment of immunodepression or a disorder involving immunodepression. Examples of disorders involving immunodepression include AIDS, cancer, senile dementia, trauma (including wound healing, surgery and shock) , chronic bacterial infection, and certain central nervous system disorders. The immunodepression to be treated may be caused by an overdose of an immunosuppressive macrocyclic compound, for example derivatives of 12-(2-cyclohexyl-l-methylvinyl)- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' ]octacos-18-ene such as FR-900506, or rapamycin. Overdosing of such medicaments by patients is quite common upon their realising that they have forgotten to take their medication at the prescribed time, and can lead to serious side effects. A further 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 for the acquisition of immunity from disease acts as an immunosuppressive agent, and so antibodies are not produced by the body and immunity is not acquired. By introducing a compound of formula I into the body (for example in the vaccine) the undesired immunosuppression may be overcome and immunity acquired. The present invention further provides the novel compounds of formula I, as defined above, provided that i) when R 1 represents OH; R 2 represents H; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H; R 9 represents OCH 3 ; X and Y each represent O; and n represents 2; then R 3 does not represent 2-oxopropyl, 2,3-dihydroxypropyl or ethanalyl; ii) when R 1 represents OH; R 2 represents H; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents OH; R 9 represents OCH 3 ; X and Y each represent O; and n represents 2; then R 3 does not represent allyl or l-hydroxyprop-2-enyl; and iii) when R~ represents OH; R 2 represents H; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H; R 9 represents OCH 3 ; X and Y each represent (H,OH) ; and n represents 2; then R 3 does not represent allyl. In the novel use or the novel compounds, we prefer R 3 to be ethyl substituted by 0 or propyl substituted by 0 - Desirably, R 7 is OH. We prefer R 2 to be OH, more preferably (S)-OH (ie to have S absolute stereochemistry at its point of attachment to the molecule) . When R 3 comprises an ester or amide group, we prefer the alcohol or amine moiety to contain from 1 to 10 carbon atoms, for example the alcohol moiety may be methanol. 5 Known compounds of formula I (as first defined above) from WO 89/05304 include: 17-Allyl-l,14,20-trihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo 0 [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-(l-Hydroxyprop-2-enyl)-1,14,20-trihydroxy-12-[2- (4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25- dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 5 17-(2,3-Dihydroxypropyl)-l,14-dihydroxy-12-[2- (4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25- dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-Ethanalyl-l,14-dihydroxy-12-[2-(4-hydroxy-3- 0 methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-(2-Oxopropyl)-l,14-dihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 25 13 ,19,21, 7-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-Allyl-l,2,14,16-tetrahydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-3,10-dione. According to the invention there is further provided a process for the production of a novel compound of formula I, which comprises: a) producing a compound of formula I, in which R 3 represents propyl substituted by 0, by oxidation of a corresponding compound in which represents allyl; b) producing a compound of formula I, which contains a vicinal diol, by oxidation of a carbon-carbon double bond in a corresponding compound; c) producing a compound of formula I, in which R 3 represents ethyl substituted by O, by oxidative cleavage of a corresponding compound in which R 3 represents 2,3-dihydroxypropy1; d) producing a compound of formula I, in which R 3 represents methyl substituted by -C0 2 H or ethyl substituted by -C0 2 H, by oxidation of a corresponding compound in which R 3 represents ethanalyl or propanalyl; e) producing a compound of formula I, which contains two vicinal hydrogen atoms, by reduction of a corresponding compound which contains a carbon-carbon double bond; f) producing a compound of formula I, in which X or Y represents (H,0H) , by reduction of a corresponding compound in which X or Y represents O; g) producing 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 methyl-substituted furanyl ring, by the action of acid on a corresponding compound in which R 3 represents 2-oxopropyl, R 4 represents H and Y represents O; h) producing a compound of formula I, in which R 1 and R 5 together represent an oxygen atom and R 6 and R 7 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, by the action of acid on a corresponding compound in which R 1 represents OH, R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, and R 7 represents OH; i) producing a compound of formula I, in which R 7 and R 8 together represent an oxygen atom, by the action of a dehydrating agent on a corresponding compound of formula I in which R 7 represents OH and R 8 represents OCH 3 ; j) producing a compound of formula I, in which R 7 represents OH, by allylic oxidation of a corresponding compound in which R 7 represents H; or k) producing a compound of formula I, in which R 3 represents allyl substituted by hydroxy, by allylic oxidation of a corresponding compound in which R 3 represents allyl. Esters and amides of carboxylic acids that R 3 may represent may be produced by conventional methods. Where desired or necessary, hydroxy groups may be protected and deprotected using conventional protecting group chemistry [as described in "Protective Groups in Organic Chemistry", ed: J W F McOmie, Plenum Press (1973), and "Protective Groups in Organic Synthesis", T W Greene, Wiley-Interscience (1981) ] . In addition, European paten application No 184162 describes the use of protectin groups in macrocyclic compounds. In process (a) , suitable oxidizing agents include a palladium (II) halide, for example palladium (II) chloride, in conjunction with a cuprous halide, for example copper (I) chloride. Suitable solvents include those that do not adversely affect the reaction, for example dimethylformamide (DMF) and water. The reaction is preferably carried out at a temperature of from 0 to 100°C, more preferably at or around room temperature. In process (b) , suitable oxidizing agents include osmium tetroxide, potassium permanganate, and iodine in conjunction with silver acetate. Osmium tetroxide is preferably used in conjunction with a regenerating agent such as 4-methylmorpholine N-oxide. Suitable solvents include those that do not adversely affect the reaction, for example diethyl ether or tetrahydrofuran (THF) . In the case of potassium permanganate, aqueous alkaline conditions are preferred. The reaction is preferably carried out at a temperature of from 0 to 100 β C, more preferably at or around room temperature. In process (c) , suitable reagents include lead tetraacetate and phenyliodoso acetate. Suitable solvents include those that do not adversely affect the reaction, for example benzene and glacial acetic acid. The reaction is preferably carried out at a temperature of from 0 to 100°C, more preferably at or around room temperature. In process (d) , suitable oxidizing agents include sodium chlorite in conjunction with sodium hydrogen phosphate. Suitable solvents include those that do not adversely affect the reaction, for example water. The reaction is preferably carried out at a temperature of from 0 to 100"C, more preferably at or around room temperature. In process (e) , the reduction may be carries out catalytically using hydrogen. Suitable catalysts include platinum catalysts (for example platinum black), and palladium catalysts (for example palladium-on-carbon) . Suitable solvents include those that do not adversely affect the reaction, for example methanol and ethanol. The reaction may be carried out at or around room temperature. In process (f) , suitable reducing agents include borane (for example in the form of borane-ammonia complex) and sodium borohydride. Suitable solvents include those that do not adversely affect the reaction, for example diethyl ether and dichloromethane. The reaction may be carried out at or around room temperature. Where desired or necessary, the (H,OH) group may be oxidized back to O by the action of copper (II) acetate in acetic acid. In processes (g) and (h) , suitable acids include p-toluenesulphonic acid. Suitable solvents include those that do not adversely affect the reaction, for example toluene. The reaction is preferably carried out at a temperature above room temperature, for example on a steam bath. In process (i) , suitable reagents include Martin' sulphurane reagent. Suitable solvents include those tha do not adversely affect the reaction, for exampl dichloromethane. The reaction is preferably carried out a a temperature below room temperature, for example -30 β C. In processes (j) and (k) , suitable reagents include Se0 2 , preferably in the presence of ^utyl hydrogen peroxide. Suitable solvents include dichloromethane, and the reaction may be carried out at or around room temperature. The invention further provides the use of the novel compounds of formula I as pharmaceuticals, and a pharmaceutical composition comprising such a compound in association with a pharmaceutically acceptable adjuvant, diluent or carrier. For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired (eg topical, parenteral or oral) and the disease indicated. However, in general, satisfactory results are obtained when the compounds are administered at a dosage of from 0.1 to 200mg per kg of animal body weight. For man the indicated total daily dosage is in the range of from lmg to lOOmg and preferably from lOmg to 500mg, which may be administered, for example twice weekly, or in divided doses from 1 to 6 times a day or in sustained release form. Thus unit dosage forms suitable for administration, eg oesophageally, comprise from 2mg to 500mg, and preferably lmg to 500mg of the compound preferably admixed with a solid or liquid pharmaceutically acceptable diluent, carrier or adjuvant. Suitable pharmaceutical compositions for administration of compounds of formula I (as first defined above) comprise (preferably less than 80%, and more preferably less than 50% by weight) of a compound of formula I (as first defined above) in combination with a pharmaceutically acceptable adjuvant, diluent or carrier. Examples of suitable adjuvants, diluents or carriers are: for tablets, capsules and dragees - microcrystalline cellulose, calcium phosphate, diatomaceous earth, a sugar such as lactose, dextrose or mannitol, talc, stearic acid, starch, sodium bicarbonate and/or gelatin; for suppositories - natural or hardened oils or waxes; and for inhalation compositions - coarse lactose. The compound of formula I (as first defined above) is preferably in a form having a mass median diameter of from 0.01 to 10 microns. The compositions may also contain suitable preserving, stabilising and wetting agents, solubilisers, sweetening and colouring agents and flavourings. The compositions may, if desired, be formulated in sustained release form. We prefer compositions which are designed to be taken oesophageally and to release their contents in the gastrointestinal tract. We have also found that the toxicity of immunosuppressive compounds, particularly immunosuppressive macrocyclic compounds including derivatives of 12-(2-cyclohexyl-1-methylvinyl)-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene (fo example FR-900506) and rapamycin, may be reduced by administering them in association with a compound of formula I as first defined above. Thus, according to a second aspect of the invention, there is provided a pharmaceutical mixture comprising a compound of formula I (as first defined above) and an immunosuppressive compound. Preferably, the greater proportion of active ingredient in such a mixture is the compound of formula I, for example the compound of formula I may be present at a ratio of greater than 10:1 by weight, for example 99:1. The invention also provides a method of treatment of immunodepression or a disorder involving immunodepression which comprises administering a therapeutically efficacious amount of a compound of formula I, as first defined above, to a patient suffering from such a condition. The compounds of formula I (as first defined have the advantage that they are less toxic, more efficacious, are longer acting, have a broader range of activity, are more potent, produce fewer side effects, are more easily absorbed or have other useful pharmacological properties, than compounds previously used in the therapeutic fields mentioned above. The compounds of formula I (as first defined above) have a number of chiral centres and may exist in a variety of stereoisomers. The invention provides the use of all optical and stereoisomers, as well as racemic mixtures, and all optical and stereoisomers of the novel compounds of formula I per se. However, the preferred stereochemistry of various chiral carbon atoms are shown in formula la, wherein R 1 to R 8 , X, Y and n are as defined above. The invention is illustrated by the following examples. Example 1 17-(2-Oxopropyl)-l-hydroxy-12-r2-(4-hvdroxy-3- methoxycvclohexyl)-1-methylvinyl]-23.25-dimethoxy- 13.19.21.27-tetramethyl-ll.28-dioxa-4-azatricyclo r22.3.1.0 ' 9 ]octacos-18-ene-2.3.10.16-tetraone a) 17-f2-0xopropyl^-l-hvdroxy-12-r2-(4-hvdroxy-3- methoxycyclohexyl)-1-methylvinyl1-23.25-dimethoxy- 13.19.21.27-tetramethyl-ll.28-dioxa-4-azatricyclo T22.3.1.0 ' 9 1octacosa-14,18-diene-2.3.10.16-tetraone To a solution of palladium (II) chloride (25mg) and copp (I) chloride (50mg) in DMF (dimethylformamide) (6ml) a water (1ml) which had been previously oxygenated by havin air bubbled through it for 30 minutes at room temperatur was added a solution of 17-allyl-l-hydroxy-12-[2-(4- hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacosa-14,18-diene-2,3,10,16-tetraone (th compound of Example 4, WO 89/05304) (lOO g) in DMF (-2ml) The reaction mixture was then stirred and oxygenated for hours at room temperature after which it was diluted wit diethyl ether. The organic extract was then washed wit dilute aqueous hydrochloric acid (1M) and brine befor being dried (MgS0 4 ) , filtered and evaporated to give th subtitle compound as an oil. b) 17-(2-Oxopropyl)-l-hγdroxy-12-r2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl1-23.25-dimethoxy- 13.19.21.27-tetramethyl-ll.28-dioxa-4-azatricyclo r22.3.1.0 4 ' 9 1octacos-18-ene-2.3.10.16-tetraone The crude product of step (a) was then re-dissolved in dr methanol and was stirred with 10% Pd-on-carbon (lOmg) unde an atmosphere of hydrogen for 4 hours. The reactio mixture was then filtered, concentrated in vacuo an chromatographed on silica eluting with acetone/hexane [2:3] to give the title compound as a foam (84mg) . MS (FAB): 889 [M+Rb] + ; 827 [M+Na] + ; 805 [M+H] + ; 787 [M-OH] + 13 C NMR (CDC1 3 ) <5: 211.1 (C16) ; 207 (C41) ; 196.3 (C2) ; 169.1 (CIO); 166.1 (C3) ; 138.7 (C19) ; 132 (C31) ; 131 (C29) ; 121.8 (C18); 97.3 (Cl) ; 84 (C34) ; 82.4 (C12) ; 75.1 (C23); 56.2 (C9) ; 48.9 (C20) ; 47.6 (C17) ; 13.3 (C39) Example 2 2-(1.14-Dihydroxy-12- 2-(4-hydroxy-3-methoxycvclohexyl)-1- methylvinyl1-23.25-dimethoxγ-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricvclof22.3.1.0 4 ' 9 1octacos-18-ene-2.3.10.16- tetraone-17-yl>-ethanoic acid A solution of FR-900506 (1.2g), 4-methylmorpholine N-oxide (1.3g) and osmium tetroxide (0.7ml of a 4% solution in water) in THF (tetrahydrofuran) (25ml) and water (14ml) was stirred for 3.5 hours at room temperature. Solid sodium metabisulphite was then added followed by ethyl acetate and Florisil (registered trade mark) and the combined mixture was filtered through celite. The separated organic extract (after washing with saturated aqueous sodium hydrogen carbonate solution) was dried (MgS0 4 ) , filtered and evaporated in vacuo to give the crude 17-(2,3- dihydroxypropyl) compound as an oil. This was then dissolved in benzene (40ml) and lead tetraacetate (1.46g) was added. After stirring for 3 minutes at room temperature the reaction mixture was diluted with diethyl ether and was filtered, concentrated in vacuo, redissolved in diethyl ether, re-filtered and re-concentrated in vacuo to give the crude 17-(ethanalyl) compound as an oil. This was then dissolved in butanol (25ml) and 1-methyl cyclohex-1-ene (6ml) and to this was added portionwise over 10 minutes solutions of sodium chlorite (0.8g) and sodi hydrogen phosphate (0.81g) each dissolved in 5ml of wate After stirring for 0.5 hours at room temperature t reaction mixture was quenched by the addition of eth acetate. Dilute aqueous hydrochloric acid (2M) was th added and the organic extracts (after washing wit saturated aqueous sodium hydrogen carbonate solution) wer dried (MgS0 4 ) , filtered and evaporated to an oil i vacuo. Chromatography on silica eluting with diethy ether/methanol/acetic acid [190:9:1] and then furthe chromatography on silica eluting with dichloromethane/ methanol/acetic acid [170:9:1] gave the title compound as foam (402mg, 32%) . MS (FAB) : 906 [M+Rb] + 13 C NMR (CDC1 3 ) δ : 196.3 (C2) ; 177.7(C41); 168 (CIO); 164.6 (C3) ; 139.7 (C19) ; 132.7 (C29) ; 129.3 (C31) 120.6 (C18) ; 97.3 (Cl) ; 84.2 (C34) ; 70.5 (C14) ; 9.8 (C39) Example 3 2-(1.14-Dihydroxγ-12-f2-(4-hydroxy-3-methoxycyclohexyl)-l- methylvinyll-23.25-dimethoxy-13 f 19.21.27-tetramethyl-ll.28- dioxa-4-azatricyclof22.3.1.0 ' 9 ]octacos-18-ene-2 f 3.10.16- tetraone-17-yl)-ethanoic acid Methyl ester To a solution of the title compound of Example 2 (50mg) i dichloromethane (3ml) at room temperature was added solution of diazomethane in diethyl ether until no startin material remained. The reaction mixture was the concentrated in vacuo and chromatographed on silica elutin with acetone/hexane [1:2] to give the title compound as foam (48mg) . S (FAB): 920 [M+Rb] + ; 858 [M+Na] + ; 818 [M-OH] + 13 C NMR (CDC1 3 ) δ : 212.7 (C16) ; 196 (C2) ; 173 (C41) ; 168.8 (CIO); 164.6 (C3) ; 139.5 (C19) ; 132.9 (C29) ; 129.1 (C31) ; 120.6 (C18) ; 97.2 (Cl) ; 84.2 (C34) ; 70.8 (C14) ; 14.5 (C30) ; 9.7 (C39) Example 4 2-11.14-Dihydroxy-12-f2-(4-hydroxy-3-methoxycyclohexyl)-1- methylvinyn-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricyclof22.3.1.0 ' 9 1octacos-18-ene-2.3.10.16- tetraone-17-yll-ethanoic acid N-Morpholine amide A solution of the title compound of Example 2 (63.1mg), morpholine (20mg) , l-(3-dimethylaminopropyl)-3- ethylcarbodii ide hydrochloride (28mg) and 4-dimethylaminopyridine (2mg) in dry dichloromethane (4ml) was stirred at room temperature for 2.5 hours. Water was then added and the reaction mixture was extracted with dichloromethane. After washing with dilute aqueous hydrochloric acid (1M) and saturated aqueous sodium hydrogen carbonate solution, the dichloromethane extracts were dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/acetone [1:1] then gave the title compound as a foam (58mg, 84%) . MS (FAB): 975 [M+Rb] + ; 891 [M+H] + ; 873 [M-0H] + . 13 C NMR (CDC1 3 ) δ : 213.2 (C16) ; 195.9 (C2) ; 170.1 (C41) ; 168.8 (CIO); 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 (CO of morpholine) ; 10.1 (C39) . Example 5 2-(l P 14-Dihγdroxy-12-r2-(4-hydroxy-3-methoxγcyclohexyl)-1- methylvinyll-23.25-dimethoxγ-13.19.21.27-tetramethyl-ll.28- 5 dιoxa-4-azatricγclor22.3♦1.0 ' J ]octacos-18-ene-2.3.10.16- tetraone-17-yl)-ethanoic acid N-Ethanolamide A solution of the title compound of Example 2 (22.8mg) and 1,3-dic/clohexyl carbodiimide (6.7mg) in dry dichloromethane (2ml) was stirred at room temperature for 5 10 minutes. Ethanolamine (lOmg) was then added and the reaction mixture was stirred at room temperature for 1.5 hours. Dilute aqueous hydrochloric acid (1M) was then added and the reaction mixture was extracted with dichloromethane. The organic extracts, after washing with 15 saturated aqueous sodium hydrogen carbonate solution, were dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with dichloromethane/methanol/acetic acid [140:9:1] then gave the title compound as a glass (19.4mg, 81%). 20 MS (FAB): 950 [M+Rb] + ; 888 [M+Na] + ; 866 [M+H] + ; 848 [M-0H] + 13 C NMR (CDC1 3 ) δ : 198.7 (C2) ; 173.8 (C41) ; 169. (CIO); 16".6 (C3) ; 140 (C19) ; 133.2 (C29) ; 129 (C31) ; 122.4 (C18) ; 97.8 (Cl) ; 84.2 (C34) ; 61.2 (CO of ethanolamine) ; 25 44.0 (CN of ethanolamine); 16.2 (C47) ; 15.8 (C43) ; 14.7 (C30) ; 9.7 (C39) Example 6 2-(1.14-Dihvdroxy-12-T2-(4-hvdroxy-3-methoxycvclohexγl)-1- methylvinyl]-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricyclof22.3.1.0 4 ' 9 ]octacos-18-ene-2.3.10.16- tetraone-17-yl)-ethanoic acid amide with σlvcine methyl ester To a solution of the title compound of Example 2 (43.6mg) and methyl glycinate (3mg) in dichloromethane (3ml) was added triethylamine (22μl) and 2-chloro-l- methylpyridinium tosylate (24.2mg). After stirring at room temperature for 15 minutes dilute aqueous hydrochloric acid (2M) was added and the reaction mixture was extracted with dichloromethane. The organic extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, were then dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/ acetone [1:1] then gave the title compound as a foam (34.1mg, 72%). MS (FAB): 977 [M+Rb] + ; 915 [M+Na] + ; 893 [M+H] + ; 875 [M-OH] + 13 C NMR (CDC1 3 ) 6 : 213 (C16) ; 195.9 (C2) ; 171.7 (glycine carbonyl) ;170.3 (C41) ; 168.7 (CIO); 164.7 (C3) ; 139.2 (C19) ; 132.9 (C29) ; 128.9 (C31) ; 120.9 (C18) ; 97.2 (Cl) ; 84.1 (C34) ; 72.4 (C24) ; 71.1 (C14) ; 52.4 (OCH3 of glycine); 9.9 (C39) Example 7 2-{1.14-Dihvdroxγ-12-r2-(4-hvdroxy-3-methoxycyclohexγl)-l- methylvinyl1-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricγcloT22.3.1.0 4 ' 9 ]octacos-18-ene-2.3.10.16- tetraone-17-yl)-ethanoic acid N-Piperidine amide To a solution of 2-chloro-l-methylpyridinium tosylat (24.9mg) in dry dichloromethane (1ml) under nitrogen a room temperature was added a solution of the title compoun of Example 2 (42.3mg), piperidine (lOmg) and triethylamin (22μl) in dry dichloromethane (1.5ml). After 3 hours a room temperature a further portion of 2-chloro-l- methylpyridinium tosylate (15mg) and triethylamine (15μl) was added and stirring was continued for further two hours at room temperature. Water was then added and the reaction mixture was extracted with dichloromethane. After washing with dilute aqueous hydrochloric acid (1M) and saturated aqueous sodium hydrogen carbonate solution, the dichloromethane extracts were dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with dichloromethane/methanol [29:1] then gave the title compound as a foam (13.3 mg, 29%). MS (FAB): 973 [M+Rb] + ; 911 [M+Na] + ; 889 [M+H] + 13 C TMR (CDC1 3 ) δ : 213.4 (C16) ; 196 (C2) ; 169. (CIO); 168.6 (C41) ; 164.7 (C3) ; 138.8 (C19) ; 133.3 (C29) ; 128.7 (C.31) ; 121.7 (C18) ; 97.3 (Cl) ; 84.1 (C34) ; 10.1 (C39) Example 8 2-(l.14-Dihγdroxy-12-r2-(4-hvdroxγ-3-methoxycyclohexyl)-l- methylvinyl]-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricyclof22.3.1.0 4 ' ]octacos-18-ene-2.3.10.16- tetraone-17-yl>-ethanoic acid N-Benzylamide A solution of the title compound of Example 2 (48mg) , 2-chloro-l-methylpyridinium tosylate (27.lmg), triethylamine (25μl) and benzylamine (lOmg) in THF (3ml) was stirred at room temperature for 1.5 hours. Dilute aqueous hydrochloric acid (2M) was then added and the mixture was extracted with ethyl acetate. The organic extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, were dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/acetone [3:2] then gave the title compound as a foam (33.6 mg, 63%). MS (FAB): 995 [M+Rb] + ; 933 [M+Na] + ; 911 [M+H] + ; 893 [M-OH] + 13 C NMR (CDC1 3 ) δ : 213.3 (C16) ; 196.1 (C2) ; 171.6 (C41) ; 168.9 (CIO); 164.9 (C3) ; 139.3 (C19) ; 133.2 (C29) ; 128.9 (C ar ) ; 128 (C ar ) ; 121.4 (C18) ; 97.5 (Cl) ; 84.3 (C34) ; 10.2 (C39) Example 9 2-(1.14-Dihvdroxγ-12-r2-(4-hvdroxy-3-methoxycvclohexyl)-1- methylvinyll-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricγclor22.3.1.0 4 ' 9 ]octacos-18-ene-2.3.10.16- tetraone-17-γl>-ethanoic acid N-Butylamide A solution of the title compound of Example 2 (57.8mg), 2-chloro-l-methylpyridinium tosylate (37.8mg), triethylamine (11.5μl) and butylamine (12mg) in THF (2ml) was stirred at room temperature for 3 hours. A further portion of 2-chloro-l-methylpyridinium tosylate (37.8mg) and triethylamine (11.5μl) were then added and stirring was continued for an additional 2 hours at room temperature, after which dilute aqueous hydrochloric acid (2M) was added and the organic extracts (after washing wit saturated aqueous sodium hydrogen carbonate solution) wer dried (MgS0 4 ) , filtered and evaporated to an oil i vacuo. Chromatography on silica eluting wit dichloromethane/methanol [29:1] then gave the titl compound as a foam (36.3mg, 59%). MS (FAB): 961 [M+Rb] + ; 899 [M+Na] + ; 877 [M+H] + ; 85 [M-OH] + 13 C NMR (CDC1 3 ) δ : 213.4 (C16) ; 196.1 (C2) ; 171. (C41) ; 168.9 (CIO); 165 (C3) ; 139.2 (C19) ; 133.3 (C29) ; 129.2 (C31) ; 121.6 (C18) ; 97.5 (Cl) ; 84.4 (C34) ; 10.2 (C39) Example 10 2-(l.14-Dihydroxy-12-r2-(4-hydroxy-3-methoxycyclohexγl)-l- methylvinyl]-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricγclo[22.3.1.0 4 ' 9 ]octacos-18-ene-2.3.10.16- tetraone-l7-yl)-ethanoic acid p-Cvanophenγl ester A solution of the title compound of Example 2 (51mg) , 2-chloro-l-methylpyridinium tosylate (33.4mg), triethylamine (29μ) and p-cyanophenol (10.2mg) in TH (2ml) was stirred at room temperature for 2 hours. Dilute aqueous hydrochloric acid (2M) was then added and the organic extracts (after washing with saturated aqueous sodium hydrogen carbonate solution) were dried (MgS0 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with dichloromethane/methanol [39:1] then gave the title compound as a foam (24.9 mg, 43%). MS (FAB): 1007 [M+Rb] + , 905 [M-OH] + 13 C NMR (CDC1 3 ) δ : 212.4 (C16) , 196 (C2) , 170. (C41) , 168.8 (CIO), 164.4 (C3), 153.8 (0C ar ) , 140.4 (C19) , 133.6 (C ar ) , 132.7 (C29) , 129.1 (C31) , 122.6 (C ar ) , 120.1 (C18) , 118.2 (CN of p-cyanophenol) , 109.7 (C ar CN) , 97 (Cl) , 84.1 (C34) , 14.4 (C30) , 9.9 (C39) 5 Example 11 2-f1.14-Dihγdroxy-12-r2-f4-hvdroxγ-3-methoxycvclohexyl)-1- methylvinyl]-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricvclor22.3.1.0 /9 1octacos-18-ene-2.3.10.16- tetraone-17-yll-ethanoic acid Phenyl ester 0 A solution of the title compound of Example 2 (45mg) , 2-chloro-l-methylpyridinium tosylate (32.3mg), triethylamine (30μl) and phenol (28mg) in dry dichloromethane (2ml) was stirred at room temperature for 2 hours. Dilute aqueous hydrochloric acid (1M) was then 5 added and the organic extracts (after washing with saturated aqueous sodium hydrogen carbonate solution) were dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/ acetone [3:2] then gave the title compound as a foam 0 (17.6mg, 36%). MS (FAB); 982 [M+Rb] + ; 920 [M+Na] + ; 898 [M+H] + ; 880 [M-0H] + 13 C NMR (CDC1 3 ) δ : 212.4 (C16) ; 195.9 (C2) ; 171.1 (C41) ; 168.8 (CIO); 164.5 (C3) ; 150.5 (0C ar ) ; 140 (C19) ; 25 132.7 (C29);- 129.4 (C ar ) ; 129.3 (C31) ; 125.8 (C ar ) ; 121.4 (C ar ) ; 120.7 (C18) ; 97.2 (Cl) ; 84.1 (C34) ; 9.7 (C39) Example 12 2-(1.14-Dihydroxy-12-r2-(4-hydroxγ-3-methoxycvclohexyl)-1- methylvinyl]-23.25-dimethoxy-13.19.21.27-tetramethyl-ll.28- dioxa-4-azatricγclor22.3.1.0 4 ' 9 1octacos-18-ene-2.3.10.16- tetraone-17-yl)-ethanoic acid p-Nitrophenyl ester 5 A solution of the title compound of Example 2 (150mg) , 2-chloro-l-methylpyridinium tosylate (77mg) , triethylamin (77μl) and p-nitrophenol (47mg) in dry dichlorometha (2ml) was stirred at room temperature for 2.5 hours. Th reaction mixture was then concentrated in vacuo an 10 purified directly by chromatography on silica eluting with hexane/diethyl ether [3:2] to give the title compound as a foam (135mg, 78%) . MS (FAB): 1027 [M+Rb] + ; 925 [M-0H] + 13 C NMR (CDC1 3 ) δ : 212.6 (C16) ; 196.2 (C2) ; 170. 15 (C41) ; 169 (CIO); 164.7 (C3) ; 155.4 (OC ar ) ; 145.5 (NC ar ) ; 140.6 (C19) ; 132.9 (C29) ; 129.3 (C31) ; 125.4 (C ar ) ; 122.6 (C ar ) ; 120.3 (C18) ; 97.2 (Cl) ; 84.3 (C34); 9.8 (C39) Example 13 2017-Allyl-1.14.16-trihvdroxy-12-r2-f4-hvdroxy-3- methoxycyclohexyl)-1-methylvinyl1-23 f 25-dimethoxy- 13.19.21 f 27-tetramethyl-ll.28-dioxa-4-azatricyclo r22.3.1.0 4,9 ]octacos-18-ene-2 f 3,10-trione To a solution of FR-900506 (lOOmg) in diethyl 25 ether/dichloromethane was added excess borane ammonia complex (150mg) . After stirring for 2 hours at room temperature dilute aqueous hydrochloric acid (IM) was added and the ethereal extract was dried (Na 2 S0 4 ) , filtered and evaporated in vacuo to give the crude 1,2,14,16- tetrahydroxy compound as an oil. This was taken up in acetic acid and copper (II) acetate (lOOmg) was added. After heating on a steam bath for 10 minutes the reaction 5 mixture was cooled to room temperature and water was added. The reaction mixture was then extracted with ethyl acetate and the organic extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, were dried (MgS0 4 ) , filtered and evaporated to an oil in 0 vacuo. Chromatography on silica eluting with acetone/hexane [1:2] then gave the title compound (30mg) with R-stereochemistry at C16 followed by the title compound (24mg) with S-stereochemistry at C16. (16R)-Stereoisomer 5 MS (FAB): 890 [M+Rb] + 13 C NMR (CDC1 3 ) δ : 199.4 (C2) ; 169.1 (CIO); 165.8 (C3) ; 136.9 (C41) ; 136.3 (C19) ; 132.6 (C29) ; 128.2 (C31) ; 125.4 (C18) ; 115.9 (C42) ; 98.7 (Cl) ; 84.2 (C34) ; 56.5 (C9) ; 49 (C20) ; 43.2 (C17) ; 10.4 (C39) 0 (16S)-Stereoisomer MS (FAB): 890 [M+Rb] + ; 828 [M+Na] + ; 806 [M+H] + ; 788 [M-OH] + 13 C NMR (CDC1 3 ) δ 196.5 (C2) ; 169.3 (CIO); 165.3 (C3) ; 137.8 (C41) ; 136.2 (C19) ; 132.9 (C29) ; 128.5 (C31) ; 25 126.6 (C18) ; 115.7 (C42) ,* 97 (Cl) ; 84.3 (C34) ; 56.4 (C9) ; 9 (C39) Example 14 1.14-Dihvdroxy-12-r2-f4-hvdroxy-3-methoxycvclohexyl)-l- methylvinyl1-26.28-dimeth_)_>rv-13.18.22.24,30-pentamethyl- 11.17.31-trioxa-4-azatetracvclor25.3.1.0 4 ' 9 .0 16 ' 20 1 hentriaconta-16 (20) .18.21-triene-2.3.10-trione A solution of 1,14-dihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-17-(2- oxopropyl)-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricycl [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone (th compound of Example 29, WO 89/05304) (lOOmg) in dr dichloromethane (25ml) containing p-toluenesulphonic acid (5mg) was heated under reflux for 30 minutes. Volatiles were then removed in vacuo and the residue was chromatographed on silica eluting with acetone/hexane [3:2] to give the title compound (19mg) as a foam. MS (FAB): 887 [M+Rb] + ; 825 [M+Na] + ;803 [M+H] + ; 785 [M-OH] + 13 C NMR (CDC1 3 ) δ i 195.9 (C2) ; 168.9 (CIO); 164. (C3); 150.8 (C16) ; 148.3 (C18) ; 106.3 (C19) ; 97 (Cl) ; 84.1 (C37); 8.5 (C42) Example 15 1.14-dihγdroxy-12-r2-(4-hydroxy-3-methoxycyclohexγl)-1- methylvinyll-23.25-dimethoxy-17-propanalyl-13.19.21.27- tetramethyl-ll.28-dioxa-4-azatricyclor22.3.1.0 4 ' 9 ]octacos- 18-ene-2.3.10.16-tetraone A solution of FR-900506 (3g) in DMF (90ml) and water (15ml) containing palladium (II) chloride (0.4g) and copper (I) chloride (l * 8g) was stirred at room temperature while air was bubbled through the reaction mixture for 2 hours. The reaction mixture was then diluted with diethyl ether and the organic extract (after washing with dilute aqueous hydrochloric acid (IM) , water and brine) was dried (MgS0 4 ) , filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane in an 5 increasing acetone gradient then gave the title compound as a foam (30mg) . MS (FAB) : 821 [M+H] 13 C NMR (CDC1 3 ) δ : 213.1 (C16) , 202.1 (C42) , 196.3 (C2) , 169 (CIO), 164.7 (C3), 139.8 (C19) , 132.6 (C29) , 10 129.6 (C31) , 121.9 (C18) , 97.1 (Cl) , 84.2 (C34) , 70.3 (C14) , 9.6 (C39) Example 16 17-Allyl-l-hydroxy-12-r2-(4-hγdroxy-3-methoxycvclohexyl) -1- methylvinyll-23.25-dimethoxγ-13.19.21.27-tetramethyl- 15 11.28.29-trioxa-4-azatetracvcloT22.3.1.1 14 ' 18 .0 4 ' 9 1 nonacos-19-ene-2.3.10.16-tetraone A sample of 17-allyl-l,14,20-trihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo 20 [22.3.1.0 ' 9 ]octacos-18-ene-2,3,10,16-tetraone (as prepared in Example 13 of WO 89/05304, 60mg) was heated in toluene (10ml) containing p-toluenesulphonic acid for 10 minutes. Evaporation of the solvent in vacuo and chromatography on silica then gave the title compound as an 25 oil (30mg) . 13 C NMR (CDC1 3 ) δ : (single rotamer) 210.22 (C16) ; 195.96 (C2); 168.96 (CIO); 165.24 (C3) ; 135.92 (C20) ; 134.69 (C41) ; 131.13 (C29) ; 128.78, 128.65 (C19, C31) ; 117.60 (C42); 97.53 (Cl) ; 84.19 (C34) ; 80.79 (C18) ; 73.167 (C14) ; 52.47 (C17) ; 44.638 (C15) ; 29.992 (C21) MS (FAB): 802.47 [M+H] + ; 886.2 [M+Rb] + -Example 17 17-Propyl-l-hydroxy-12-r2-(4-hydroxγ-3-methoxycγclohexyl)-1- ethylvinyl]-25-dimethoxγ-13.19.21.27-tetramethyl-ll.28.29- trioxa-4-azatetracyclor22.3.1.1 20 ' 23 .0 ' 9 ]nonacos-18-ene -2.3.10.16-tetraone A sample of 17-allyl-l,14,20-trihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone (as prepared in Example 13 of WO 89/05304, 121mg) was dissolved in dry dichloromethane (10ml) and to this was added Martin's sulphurane reagent at -30 β C. After warming to 0°C, volatiles were removed in vacuo and the residue was chromatographed on silica to give a foam. To a solution of this in dry methanol (5ml) was added 10% Pd-on-carbon (lOmg) and the resulting suspension was stirred in an atmosphere of hydrogen for 7 hours at room temperature. Volatiles were then removed in vacuo and the residue was purified by column chromatography on silica to yield the title compound as an oil (19mg). MS (FAB): 770.76 [M+H] + , 792.74 [M+Na] + , 854.44 [M+Rb] + 13 C NMR (CDC1 3 ) δ (single rotamer) 197.81 (C16) ; 196.11 (C2); 168.96 (CIO); 165.21 (C3) ; 146.16 (C14) ; 139.46 (C19) ; 135.92 (C41) ; 134.63 (C31) ; 129.51 (C29) ; 129.40 (C15) ; 125.31 (C18) ; 116.29 (C42) ; 97.33 (Cl) ; 92.07 (C20) ; 84.09 (C34) ; 81.75 (C12) ; 80.03 (C23); 77.10 (C25) ; 74.67 (C24) ; 73.43 (C35) ; 56.78 (C9) ; 56.47 (OCH 3 ); 55.95 (OCH 3 ) ; 51.37 (C17) 5 Example 18 17-Allyl-l-hydroxy-12-r2-(4-hydroxy-3-methoxycyclohexyl)-1- methylvinyn-25-methoxy-13.19.21.27-tetramethyl-ll,28,29- trioxa-4-azatetracyclor22.3.1.1 20 ' 23 .0 4 ' 9 1nonacosa- 14.18-diene-2.3,10.16-tetraone 0 A sample of 17-allyl-l,14,20-trihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone (as prepared in Example 13 of WO 89/05304, 80mg) was dissolved 5 in dry dichloromethane (5ml) and to this was added excess Martin's sulphurane reagent at -30 β C portionwise until all the starting material had disappeared (30 minutes) . The reaction mixture was then diluted with ethyl acetate and washed with water and then brine. The organic extract was 20 dried (MgS0 4 ) , filtered and evaporated in vacuo to an oil. Chromatography on silica then gave the title compound as an oil (20mg) . 13 C NMR (CDC1 3 ) 6: (mixture of rotamers) 212.05, 211.04 (C16) ; 197.47, 194.95 (C2) ; 169.24, 169.09 (CIO); 25165.21, 164.72 (C3) ; 136.85, 137.44 (C19) ; 131.43, 130.97 (C29) ; 131.25, 128.90 (C31) ; 125.02, 124.35 (C18) ; 98.82, 97.44 (Cl) ; 91.56, 91.29 (C20) ; 84.17 (C34) MS (FAB): 775.11 [M+H] + ; 797.09 [M+Na] + ; 858.79 [M+Rb] + Example 19 12-f2-(4-Hydroxy-3-methoxycyclohexyl)-1-methylvinyll-23.25- dimethoxy-17-propyl-13.19.21.27-tetramethγl-l.14.15- trihγdroxy-11.28-dioxa-4-azatricyclof22.3.1.0 4 ' 9 J. octacos-18-ene-2.3.10.16-tetraone A sample of l-hydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-17-propyl-13,19,21,27- tetramethyl-ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacosa -14,18-diene-2,3,10,16-tetraone as an oil (the compound o Example 11 of WO 89/05304, 510mg) was dissolved in dr tetrahydrofuran (12ml) and N-methyl morpholine N-oxid (0.3g) and osmium tetroxide (3ml of a 4% aqueous solution) were added. After stirring for 1 hour at room temperatur solid sodium metabisulphite (lg) was added followed after 5 minutes by celite to produce a thick slurry. The reactio mixture was then diluted with ethyl acetate and filtered. The organic extract after washing with saturated aqueous sodium bicarbonate solution and brine was dried (magnesiu sulphate), filtered and concentrated in vacuo to an oil. Column chromatogarphy on silica eluting with acetone/hexane [2:5] give two isomers of the title compound which differed in stereochemistry at C14 and/or C15. Isomer 1 (180mg) 13 C NMR δ z (For the major rotamer) 211.82 (C16) 196.03 (C2); 169.10 (CIO); 164.26 (C3) ; 139.28 (C19) ; 132.12 (C29) ; 128.22 (C31) ; 121.92 (C18) ; 95.65 (Cl) ; 83.94 (C34); 75.76 (C12); 74.81 (C23) ; 74.63 (C15) ; 73.26 (C25) ; 73.26 (C35) ; 72.22 (C24) ; 71.66 (C14) ; 56.02 (C9) ; 48.05 (C20) ; 47.29 (C17) ; 39.08 (C5) ; 37.90 (C13) ; 34.92 (C33) ; 34.69 (C32) ; 34.47 (C27);33.66 (C22) ; 32.78 (C40) ; 32.63 (C26) ; 31.14 (C36) ; 30.57 (C37) ; 28.66 (C8) ; 25.78 (C21) ; 5 24.47 (C6) ; 21.24 (C7) ; 19.83 (C41) ; 19.83 (C44) ; 16.08 (C47) ; 15.66 (C43) ; 14.68 (C30) ; 14.07 (C42) ; 8.62 (C39) X H NMR δ : (For the Major rotamer) 5.48 (lH,brs,H-12) ; 5.15 (lH,d,J=9.6Hz,H-18) ; 5.08 (lH,d, =8.8Hz,H-31) ; 3.79 (lH,dd,J=1.5 and 8Hz,H-24) ; 1.66 (3H,brs,H-30) ; 1.60 0 (3H,brs,H-43) ; 1.02 (3H,d,J=6.5Hz,H-47) ; 0.92 (3H,t,J=7.6 Hz,H-42); 0.90 (3H,d,J=7.6Hz,H-44) ; 0.83 (3H,d,J=7 Hz,H-39) MS: 823 [M+H] + ; 845 [M+Na] + ; 906 [M+Rb] + Isomer 2 (32mg) 13 C NMR δ : (For the major rotamer) 212.99 (C16) ; 15 194.39 (C2); 169.60 (CIO); 164.85 (C3) ; 138.33 (C19) ; 131.44 (C29) ; 129.95 (C31) ; 123.16 (C18) ; 97.79 (Cl) ; 84.17 (C34) ; 76.54 (C12) ; 75.73 (C23) ; 75.68 (C15) ; 73.52 (C25) ; 73.52 (C35) ; 72.78 (C24) ; 71.88 (C14) ; 55.85 (C9) ; 49.21 (C17) ; 48.78 (C20) ; 39.36 (C5) ; 38.83 (C13) ; 36.04 (C40) ; 20 35.34 (C32) ; 34.83 (C33) ; 34.46 (C27) ; 33.21 (C22) ; 32.35 (C26) ; 31.21 (C36) ; 30.58 (C37) ; 28.34 (C8) ; 26.33 (C21) ; 24.75 (C6) ; 20.76 (C41) ; 20.63 (C7) ; 20.42 (C44) ; 16.31 (C47) ; 15.31 (C43) ; 14.17 (C30) ; 13.95 (C42) ; 9.64 (C39) . 1 H NMR δ : (For the major rotamer) 5.24 (lH,brs,H-12) ; 254.74 (lH,t,J=3.4Hz,H-9) ; 3.66 (lH,dd,J=1.4 and 9.6Hz,H-24); 1.63 (3H,brs,H-30) ; 1.57 (3H,brs,H-43) ; 0.99 (3H,d,J=6.4Hz,H-47) ; 0.94 (3H,d,J=7.3Hz,H-39) ; 0.92 (3H,d,J=4.5Hz,H-44) ; 0.90 (3H,t,J=7.5 Hz,H-42) MS: 823 [M+H] + ; 845 [M+Na] + ; 906 [M+Rb] + Example 20 In the mixed lymphocyte reaction (MLR) (described in W 89/05304, Example A) , 17-(2-Oxopropyl)-l,14-dihydroxy- 12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25- dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4,9 ]octacos-18-ene-2,3,10,16-tetraone, was found to have a pA 2 value of 8.3 with agonist compound FR-900506. This means that a concentration of 5xlO~ 9 M of the antagonist compound is required to occupy 50% of the available receptor sites in the screen for which it competes with FR-900506.

Claims: CLAIMS : 1. The use of a compound of formula I, wherein R 1 and R 2 independently represent H or OH, or they may together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 3 represents methyl optionally substituted by -C0 2 H or an ester or amide thereof; ethyl optionally substituted by O, OH or -C0 2 H or an ester or amide thereof; propyl optionally substituted by OH or 0; or allyl optionally substituted by OH; R 4 represents H; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H or OH; R 8 represents OCH 3 ; R 9 represents OH or OCH 3 ; X represents 0 or (H,OH) ; Y represents O or (H,OH) ; and n represents 1 or 2; in addition to their significances above R 1 and R 5 may together represent an oxygen atom, in which case R 6 and R 7 together represent a secon carbon-carbon bond between the carbon atoms to which the are attached; R 7 and R 8 may together represent an oxygen atom; and R 3 , R 4 and Y, together with the carbon atoms to which they are attached, may represent a methyl-substituted furanyl ring; provided that i) when R 2 represents H; R 3 represents methyl, ethyl, propyl or allyl; R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 8 represents OCH 3 ; and Y represents O; then R 7 represents OH; and ii) when n is 1, then R 3 is not methyl or ethyl; in the manufacture of a medicament for the treatment of immunodepression or a disorder involving immunodepression. 2. A compound of formula I, as defined in claim 1, provided that i) when R 1 represents OH; R 2 represents H; R 5 and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H; R 9 represents OCH 3 ; X and Y each represent O; and n represents 2; then R 3 does not represent 2-oxopropyl, 2,3-dihydroxypropyl or ethanalyl; ii) when R 1 represents OH; R 2 represents H; R 5 and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached.; R 7 represents OH; R 9 represents OCH 3 ; X and Y each represent O; and n represents 2; then R 3 does not represent allyl or l-hydroxyprop-2-enyl; and iii) when R 1 represents OH; R 2 represents H; R 5 and R together represent a second carbon-carbon bond between the carbon atoms to which they are attached; R 7 represents H; R 9 represents OCH 3 ; X and Y each represent (H,OH) ; and n represents 2; then R 3 does not represent allyl. 3. The use of a compound of formula I, as defined in claim 1, or a compound of formula I as defined in claim 2, wherein R 3 is ethyl substituted by O or propyl substituted by O. 4. The use of a compound of formula I, as defined in claim 1, or a compound of formula I as defined in claim 2, wherein R 7 is OH. 5. The use of a compound of formula I, as defined in claim 1, or a compound of formula I as defined in claim 2, wherein R 2 is OH. 6. The use of a compound of formula I, as defined i claim 1, wherein the compound of formula I is: 17-Allyl-l,14,20-trihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-(l-Hydroxyprop-2-enyl)-1,14,20-trihydroxy-12-[2- (4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25- dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 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-13,19,21,27-tetramethyl-ll ,28-dioxa-4-azatricyclo [22.3.1.0 4 *9 ]octacos-18-ene-2,3,10,16-tetraone, 17-Ethanalyl-l,14-dihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 17-(2-0xopropyl)-l,14-dihydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, or 17-Allyl-l,2,1 ,16-tetrahydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-3 ,10-dione. 7. A compound of formula I, as defined in claim 2, which s: 17-(2-Oxopropyl)-l-hydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo 5 [22.3.1.0 4 ' 9 ]octacosa-14,18-diene-2,3,10,16-tetraone, 17-(2-Oxopropyl)-l-hydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone, 0 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid, 2-{1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) 5 -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid Methyl ester, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- 20 11,28-dioxa-4-azatricyclo[22.3.1.0 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid N-Morpholine amide, 2-(1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 252,3,10,16-tetraone-17-yl}-ethanoic acid N-Ethanolamide, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid amide with glycin methyl ester, 2-{1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- 11,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid N-Piperidine amide, 2-(l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl)-ethanoic acid N-Benzylamide, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl)-ethanoic acid N-Butylamide, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl}-ethanoic acid p-Cyanophenyl ester, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl)-ethanoic acid Phenyl ester, 2-{l,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl) -1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl- ll ,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene- 2,3,10,16-tetraone-17-yl)-ethanoic acid p-Nitrophenyl ester, 17-Allyl-l,14,16-trihydroxy-l2-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10-trione 1,14-Dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1- methylvinyl]-26,28-dimethoxy-13,18,22,24,30-pentamethyl- 11,17,3l-trioxa-4-azatetracyclo[25.3.1.0 4 ' 9 .0 16 ' 20 ] hentriaconta-16(20) ,18,21-triene-2,3,10-trione, 1,14-dihydroxy-12-[2-( -hydroxy-3-methoxycyclohexyl)-1- methylvinyl]-23,25-dimethoxy-17-propanalyl-13,19,21,27- tetramethyl-ll,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos- 18-ene-2,3,10,16-tetraone, 17-Allyl-l-hydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy- 13,19,21,27-tetramethyl-ll,28,29-trioxa-4-azatetracyclo [22.3.1.I 14 ' 18 .0 4 ' 9 ]nonacos-19-ene-2,3,10,16-tetraone, 17-Propyl-l-hydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-25-dimethoxy-13,19,21,27- tetramethyl-ll,28,29-trioxa-4-azatetracyclo[22.3.1.1 20 ' 23 . ° ' ]nonacos-18-ene-2,3,10,16-tetraone, 17-Allyl-l-hydroxy-12-[2-(4-hydroxy-3- methoxycyclohexyl)-1-methylvinyl]-25-methoxy-13,19,21,27- tetramethyl-ll,28,29-trioxa-4-azatetracyclo[22.3.1.I 20 ' 23 . 0 4 ' 9 ]nonacosa-14,18-diene-2,3,10,16-tetraone, or 12-[2-(4-Hydroxy-3-methoxycyclohexyl)-1-methylvinyl]- 23,25-dimethoxy-17-propyl-13,19,21 ,27-tetramethyl-l,14,15- trihydroxy-11,28-dioxa-4-azatricyclo[22.3.1.0 ' 9 ] octacos-18-ene-2,3,10,16-tetraone. 8. The use of a compound of formula I, as defined claim 2, as a pharmaceutical. 9. A pharmaceutical composition comprising a compound o formula I, as defined in claim 2, in association with 5 pharmaceutically acceptable adjuvant, diluent or carrier. 10. A pharmaceutical mixture comprising a compound o formula I as defined in claim 1 or claim 2, and a immunosuppressive compound. 11. A process for the production of a compound of formul 0 I, as defined in claim 2, which comprises: a) producing a compound of formula I, in which R represents propyl substituted by O, by oxidation of corresponding compound in which R 3 represents allyl; b) producing a compound of formula I, which contains 5 vicinal diol, by oxidation of a carbon-carbon double bon in a corresponding compound; c) producing a compound of formula I, in which R represents ethyl substituted by O, by oxidative cleavage o a corresponding compound in which R 3 represent Q 2,3-dihydroxypropyl; d) producing a compound of formula I, in which R represents methyl substituted by ~ 0 2 H or ethy substituted by -C0 2 H, by oxidation of a correspondin compound in which R 3 represents ethanalyl or propa τ -lyl; _ e) producing a compound of formula I, which co;_ ins tw vicinal hydrogen atoms, by reduction of a correspondin compound which contains a carbon-carbon double bond; f) producing a compound of formula I, in which X or represents (H,OH) , by reduction of a corresponding compound in which X or Y represents O; g) producing 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 methyl-substituted furanyl ring, by the action of acid on a corresponding compound in which R 3 represents 2-oxopropyl, R 4 represents H and Y represents O; h) producing a compound of formula I, in which R 1 and R together represent an oxygen atom and R 6 and R 7 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, by the action of acid on a corresponding compound in which R 1 represents OH, R 5 and R 6 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, and R 7 represents OH; i) producing a compound of formula I, in which R 7 and Q R together represent an oxygen atom, by the action of a dehydrating agent on a corresponding compound of formula I in which R 7 represents OH and R 8 represents OCH 3 ; j) producing a compound of formula I, in which R 7 represents OH, by allylic oxidation of a corresponding compound in which R 7 represents H; or k) producing a compound of formula I, in which R 3 represents allyl substituted by hydroxy, by allylic oxidation of a corresponding compound in which R 3 represents allyl. 12. A method of treatment of immunodepression, or a disorder involving immunodepression, which comprise administering a therapeutically efficacious amount of compound of formula I, as defined in claim 1, to a patien suffering from such a condition. 5 10 15 20 25 4002cαnplete/jrh

Inventors: DONALD DAVID KEITH, FURBER MARK, HARDERN DAVID NORMAN, LEFF PAUL, DONALD, DAVID, KEITH, FURBER, MARK, HARDERN, DAVID, NORMAN, LEFF, PAUL
Applicants: FISONS PLC
Application year: 1990
Main classification: A61K31
IPC codes: A61K31, A61P37, C07D498, C07H19