IE910847A1 - Immunosuppressive macrocyclic compounds - Google Patents

Immunosuppressive macrocyclic compounds

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Publication number
IE910847A1
IE910847A1 IE084791A IE84791A IE910847A1 IE 910847 A1 IE910847 A1 IE 910847A1 IE 084791 A IE084791 A IE 084791A IE 84791 A IE84791 A IE 84791A IE 910847 A1 IE910847 A1 IE 910847A1
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IE
Ireland
Prior art keywords
compound
formula
producing
dioxa
tetraone
Prior art date
Application number
IE084791A
Original Assignee
Fisons Plc
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Publication date
Priority claimed from GB909005672A external-priority patent/GB9005672D0/en
Priority claimed from GB909008556A external-priority patent/GB9008556D0/en
Priority claimed from GB909008507A external-priority patent/GB9008507D0/en
Priority claimed from GB909009480A external-priority patent/GB9009480D0/en
Priority claimed from GB909017447A external-priority patent/GB9017447D0/en
Priority claimed from GB909023242A external-priority patent/GB9023242D0/en
Application filed by Fisons Plc filed Critical Fisons Plc
Publication of IE910847A1 publication Critical patent/IE910847A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/12—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains three hetero rings
    • C07D498/18—Bridged systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
    • C07H19/01—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing oxygen

Abstract

There are provided compounds of formula (I), wherein R<1> represents H, OH or alkoxy; R<2> represents H; in addition, R<1> and R<2> may together represent a second bond between the carbon atoms to which they are attached; R<3> represents methyl, ethyl, propyl or allyl; R<4> represents H, OH, alkyl, alkoxy, halogen, amino, S-alkyl, NHCHO or NHCO-alkyl; n represents 1 or 2; X represents O, (H, OH), (H, H) or =NH; and Y represents an optionally substituted cyclohexyl or substituted cyclopentyl group; with various provisos. The compounds are useful, inter alia, as immunosuppressive agents.

Description

-1 IMMUNOSUPPRESSIVE MACROCYCLIC COMPOUNDS This invention relates to immunosuppressive macrocyclic compounds, processes for their preparation, their use as medicaments, and compositions containing them.
European Patent Application 184162 (to Fujisawa Pharmaceuticals Co Ltd) discloses a number of macrocyclic compounds isolated from microorganisms belonging to the genus Streptomyces. The macrolides are numbered FR-900506, FR-900520, FR-900523 and FR-900525, and the preparation of some of their derivatives is also described.
International Patent Applications Nos WO 89/05304 and 15 PCT/GB90/01262 and European Patent Application No 413532 (to Fisons pic), European Patent Application 353678 (to Fuj isawa Pharmaceuticals Co Ltd), European Patent Applications 349049, 349061, 358508 and 388153 (to Merck & Co Inc) and European Patent Application 356399 and International Patent Application WO 90/15805 (to Sandoz AG) also disclose a number of immunosuppressive macrocyclic compounds.
We have now found a new group of immunosuppressive macrocyclic compounds which possess advantageous properties over those disclosed previously.
According to the present invention, there is provided a compound of formula I, wherein R1 represents H, OH or alkoxy; R2 represents H; in addition, R1 and R2 may together represent a second bond between the carbon atoms to which they are attached; R3 represents methyl, ethyl, propyl or allyl; R4 represents H, OH, alkyl, alkoxy, halogen, amino, 20 S-alkyl, NHCHO or NHCO-alkyl; n represents 1 or 2; X represents Ο, (H,OH), (H,H) or =NH; and Y represents a cyclic group of formula II, II (Η,ΟΗ), (H,methoxy) or 0; R° represents H, (R)-OH, (S)-OH, alkoxy, amino, 37 alkylamino, alkanoylamino, formyloxy or halogen; represents H; and in addition R5 and R6 may together represent a second bond between the carbon atoms to which 5 they are attached; or R6 and R7 may together represent a second bond between the carbon atoms to which they are attached; or a cyclic group of formula III, III in which R8 represents alkyl substituted by one or more 15 groups selected from OH, alkoxy, =0, and CO2H; or alkenyl optionally substituted by one or more groups selected from OH, =0, or CO2H; provided that a) when n represents 1; R1 represents OH; R3 2Q represents allyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent 0; b) when n represents 2; i) R1 represents OH; allyl or propyl; (H,methoxy) ; and R* represent 0; ii) when R1 and R2 together represent a second bond R3 represents methyl, ethyl, R** represents OH; R5 represents represents (R)-OH; then X does not >4 - 4 between the carbon atoms to which they are attached or each represent H; R3 represents allyl or propyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent 0; iii) when R1 represents OH, methoxy or together with R2 it represents a second bond between the carbon atoms to which they are attached; R3 represents allyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents methoxy; then X does not represent 0; iv) when R1 represents H or OH; R3 represents allyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent (H,OH); v) when R1 represents H; R3 represents propyl; R4 represents OH; R5 represents (H,OH); and R6 represents 15 (R)-OH; then X does not represent 0; vi) when R1 represents OH; R3 represents ethyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent (H,OH); vii) when R1 and R2 together represent a second bond 2q between the carbon atoms to which they are attached or each represent H; R3 represents ethyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent O; viii) when R1 represents OH; R3 represents allyl; R4 represents OH; R5 represents (H,0H) or (H,methoxy); and R6 represents (R)-OH; then X does not represent (H,H); ix) when R1 represents OH; R3 represents ethyl; R4 represents OH; R5 represents (H,methoxy); and R6 represents (R)-OH; then X does not represent (H,H); x) when R1 represents OH; R3 represents methyl, ethyl or allyl; R4 represents OH; R5 represents (H,OH); and R6 represents (R)-OH; then X does not represent 0; and xi) when R1 represents OH; R3 represents allyl; R4 represents OH; R5 represents 0; and R6 represents (R)-OH; then X does not represent 0; and pharmaceutically acceptable derivatives thereof.
Pharmaceutically acceptable derivatives which may be mentioned include esters, amides and salts of any carboxylic acid groups which may be present. The esters and amides preferably contain up to 6 carbon atoms. Salts include alkali metal and alkaline earth metal salts, for example sodium or calcium.
When any one of R1, R4, R5, R6, and R8 represent carbon-containing groups, we prefer those groups to contain up to 10 carbon atoms, more preferably up to 6 carbon 2q atoms· Groups which R8 may represent include CHO and CO2H.
Preferably, R1 represents H or OH. We prefer R4 to 25 represent H, OH, alkyl, halogen or «unino. Desirably, R5 represents (H,OH) or (H,methoxy). Preferably R6 represents H, (R)-OH or amino. We prefer R8 to represent an amide of a C02H group or alkyl substituted by alkoxy.
Subgroups of compounds which may be mentioned include: compounds of formula I in which Y represents a cyclic group of formula III; compounds of formula I in which R4 represents alkoxy; compounds of formula I in which R4 represents amino, alkylamino, alkanoylamino, halogen and thioalkyl; compounds of formula I in which R4 represents H or alkyl; and compounds of formula I in which R6 represents H, (S)-OH or halogen or together with R5 represents a second bond between the carbon atoms to which they are attached or together represent a pair of vicinal hydrogen atoms.
A preferred group of specific compounds which may be mentioned is: L5 17-allyl-l,14-dihydroxy-12-[2-(3-methoxycyclohexyl)-1methy1vinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28dioxa-4-azatricyclo[22.3.1.04·9]octacos-18-ene-2,3,10,16tetraone; 17-allyl-l,14-dihydroxy-12-[2-(cyclopentyl-3-carboxy1ic acid morpholine amide)-1-methylvinyl]-23,25-dimethoxy«u 13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone; 17-allyl-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyc1ohexy1) -1methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-ll,28dioxa-4-azatricyclo[22.3.1.04'9]octacos-18-ene-2,3,10,16tetraone; 17-allyl-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyc1ohexy1)-1methylvinyl]-23,25-dimethoxy-l,13,19,21,27-pentamethyl11.28- dioxa-4-azatricyclo[22.3.1.04·9]octacos-18-ene2.3.10.16- tetraone; 17-allyl-l-amino-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyclo hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetra methyl-11,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos-18ene-2,3,10,16-tetraone; 17-allyl-l-fluoro-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyclo hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos10 l8-ene-2,3,10,16-tetraone; 17-Ally1-1,14-dihydroxy-12-(2-(cyclopentyl-3-methanol(methyl ether))-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetra methyl-11,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos-18ene-2,3,10,16-tetraone; and 17-Allyl-l,14-dihydroxy-12-[2-(4-amino-3-methoxycyclohexyl)1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl11.28- dioxa-4-azatricyclo[22.3.1.04'9]octacos-18-ene2.3.10.16- tetraone. 2q The compounds disclosed in the above-mentioned applications may be used as starting materials for the production of compounds of the present invention. Alternatively, they may be prepared by total synthesis.
According to a further aspect of the invention, there is provided a process for the production of a compound of formula I as defined in claim 1, which comprises; (a) producing a compound of formula I in which R1 and R2 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, by dehydration of a corresponding compound in which R1 represents OH and R2 represents H; (b) producing a compound of formula I in which R1 and R2 each represent hydrogen, by reduction of a corresponding compound in which R1 and R2 together represent a second carbon-carbon bond between the carbon atoms to which they are attached; IQ (c) producing a compound of formula I in which X represents (H,OH), by reduction of a corresponding compound in which X represents 0; (d) producing a compound of formula I in which X represents (Η,Η), by reduction of a corresponding compound in which X represents 0; (e) producing a compound of formula I in which X represents 0, by oxidation of a corresponding compound in which X represents (H,0H); (f) producing a compound of formula I in which R4 2q represents alkoxy, by reaction of a corresponding compound in which R4 represents OH and X represents (H,OH) with an alkanol; (g) producing a compound of formula I in which R4 represents halogen, by reaction of a corresponding compound in which R4 represents OH with a suitable halogenating agent; (h) producing a compound of formula I in which R4 represents H or alkyl, by reaction of a corresponding compound in which R4 represents halogen with an organometallic reagent; (i) producing a compound of formula I in which R4 represents amino, by reaction of a corresponding compound 5 in which R4 represents halogen with ammonia; (j) producing a compound of formula I in which X represents =NH, by reaction of a corresponding compound in which X represents 0 with ammonia; (k) producing compound of formula I in which R' represents S-alkyl, by reaction of a corresponding compound in which R4 represents halogen with an alkylthiol; (1) producing a compound of formula I in which R4 represents NHCHO, by reaction of a corresponding compound in which R4 represents amino with formic acid; (m) producing a compound of formula I in which R4 represents NHCO-alkyl, by reaction of a corresponding compound in which R4 represents amino with an alkanoic anhydride; (n) producing a compound of formula I in which R6 represents (S)-OH, by elimination of a leaving group from a correpsonding compound in which R6 represents the leaving group; (o) producing a compound of formula I in which R8 represents H and R5 represents O, by elimination of a leaving group from a correpsonding compound in which R6 represents the leaving group; (p) producing a compound of formula I in which R6 and R7 together represent a second bond between the carbon atoms to which they are attached, by elimination of a leaving group from a correpsonding compound in which R8 represents the leaving group? (q) producing a compound of formula I in which Y represents a cyclic group of formula III and R8 represents CHO, by elimination of a leaving group from a correpsonding compound in which R8 represents the leaving group ? (r) producing a compound of formula I in which R8 Ιθ represents halogen, by reaction of a corresponding compound in which R8 represents a leaving group with halide ion? (s) producing a compound of formula I in which R5 and R8 together represent a second bond between the carbon atoms to which they are attached, by elimination of halogen and alkoxy from a corresponding compound in which R5 represents alkoxy and R8 represents halogen; (t) producing a compound of formula I in which R5 represents (H,H) and R8 represents H, by reduction of a corresponding compound in which R8 and R8 together represent a second bond between the carbon atoms to which they are attached; (u) producing a compound of formula I in which R8 represents H, by the action of hydride on a corresponding compound in which R8 represents a leaving group; (v) producing a compound of formula I in which R8 represents amino, by reduction of a corresponding compound in which R8 represents azido; (w) producing a compound of formula I in which R8 IE 91847 - 11 represents alkylamino or alkanoylamino, by reaction of a corresponding compound in which R6 represents amino with a suitable alkylating or acylating reagent; (x) producing compound of formula I in which R* represents alkyl substituted by OH, by reduction of a corresponding compound in which R8 represents alkyl substituted by =0; (y) producing compound of formula I in which includes a carboxylic acid group, by oxidation of a corresponding compound in which R8 includes an aldehyde group; and (z) producing a compound of formula I in which R8 represents optionally substituted alkenyl, by a Wittig reaction between a corresponding compound in which R8 includes an aldehyde and an appropriate Wittig reagent.
In process (a), the dehydration may be carried out in a solvent which does not adversely affect the reaction (eg toluene), in the presence of a trace amount of acid (eg p-toluenesulphonic acid), at a temperature of from 50 to 100’C.
In processes (b) and (t), the reduction may be carried out catalytically using hydrogen. Suitable catalysts include platinum catalysts (eg platinum black, platinum oxides), palladium catalysts (eg palladium oxides, palladium on charcoal), nickel catalysts (eg nickel oxide, Raney Nickel), and rhodium catalysts (eg rhodium on alumina).
Suitable solvents are those which do not adversely affect the reaction, and include methanol, ethanol, ethyl acetate, dichloromethane and dimethylformamide. The reduction may be carried out at or around room temperature.
In process (c), suitable reagents for the reduction include tri-nbutyltin hydride in a solvent which does not adversely affect the reaction (eg toluene) at a temperature of from 50 to 100°C, sodium borohydride, zinc in acetic IQ acid at or around room temperature, sodium triacetoxyborohydride in acetic acid, L-Selectride (Registered Trade Mark) in tetrahydrofuran, or borane/tbutylamine complex in a solvent such as methanol or ethanol.
In process (d), the reduction may be achieved by the action of H2S, preferably in the presence of pyridine or an amine (for example morpholine), in a solvent which does not adversely affect the reaction (for example 2q dimethylformamide, pyridine or methanol), at or around room temperature.
In process (e), the oxidation may be carried out in the presence of a suitable oxidizing agent, such as cupric acetate. Suitable solvents include those which do not adversely affect the reaction, for example methanol. The reaction may be carried out up to the reflux temperature of the solvent.
In process (f), the reaction may be carried out in the presence of a suitable acid catalyst, for example montmorilIonite K10. The solvent used may conveniently be the alkanol reagent, and the reaction may be carried out at or around room temperature.
In process (g), suitable halogenating agents include diethylaminosulphur trifluoride and thionyl chloride. The halogenation is preferably carried out in a solvent which does not adversely affect the reaction, for example dichloromethane, at or below room temperature, and preferably under an inert atmosphere.
In process (h), suitable organometallic reagents include lithium dialkyl copper reagents, which may be prepared from a copper halide and an alkyl lithium reagent. R4 preferably represents Cl in the starting material.
Suitable solvents include those which do not adversely 2q affect the reaction, for example diethyl ether. The reaction is preferably carried out at reduced temperature.
In processes (i) and (j), suitable solvents include those which do not adversely affect the reaction, for example 25 diethyl ether. R4 preferably represents Cl in the starting material. The reaction may be carried out at or around room temperature.
In process (k), suitable solvents include those which do not adversely affect the reaction, for example tetrahydrofuran (THF). R4 preferably represents Cl in the starting material. The reaction may be carried out at or around room temperature.
In process (1), the solvent is conveniently formic acid. The reaction may be carried out at or around room temperature, and in the presence of acetic anhydride.
In process (m), suitable solvents include those which do not adversely affect the reaction, for example methanol. The reaction may be carried out at below room temperature.
In processes (n)-(q), suitable leaving groups include tosylate, mesylate and triflate (trifluoromethylsulphonyloxy), and the elimination is carried out in the presence of an acid catalyst, preferably silica. The leaving group may be introduced by reaction of 2q a compound of formula I in which R8 represents (R)-OH with a suitable reagent, for example trifluoromethanesulphonic acid anhydride.
In process (r), suitable leaving groups include tosylate, 2g mesylate and triflate. Suitable sources of halide include tetra-nbutylammonium halides, for example tetra-nbutylammonium iodide. Suitable solvents include those which do not adversely affect the reaction, for - 15 Ε91847 example benzene. The reaction may be carried out at at or around room temperature.
In process (s), the elimination is preferably carried out by the action of powdered zinc. The solvent is preferably acetic acid and the reaction may be carried out at or around room temperature.
In process (u), suitable leaving groups include !θ imidazol-l-yl(thiocabonyl)oxy, which may be introduced by reaction of a corresponding compound in which R6 represents OH with 1,1'-thiocarbonyldiimidazole. Suitable sources of hydride include tributyltin hydride, and the reaction is preferably carried out in the presence of AIBN. Suitable solvents include those which do not adversely affect the reaction, for example benzene. The reaction may be carried out up to the reflux temperature of the solvent.
In process (ν), suitable reducing agents include 1,3-propanedithiol. Suitable solvents include those which do not adversely affect the reaction, for example methanol. The reaction is preferably carried out in the presence of triethylamine, and may be carried out at or 25 around room temperature. The azido compound may be produced by the action of azide ion on a corresponding compound in which R6 represents a leaving group, for example triflate.
In process (w), suitable alkylating agents include methyl iodide, and suitable acylating agents include acyl halides, for example acetyl chloride. Suitable solvents include those which do not adversely affect the reaction, for example dichloromethane. The reaction may be carried out at or around room temperature.
In process (x), suitable reducing agents include Ιθ L-Selectride. Suitable solvents include those which do not adversely affect the reaction, for example THF. The reaction is preferably carried out below room temperature. oxidizing agents include sodium in the presence of Suitable solvents include those affect the reaction, for example is preferably carried out at or In process (y), suitable chlorite, preferably 1-methylcyclohex-l-ene. which do not adversely tbutanol. The reaction around room temperature.
In process (z), suitable Wittig reagents include (carbomethoxymethylene)triphenylphosphorane. Suitable solvents include those which do not adversely affect the reaction, for example toluene. The reaction may be carried out at or around the reflux temperature of the solvent.
Conventional methods may then be used to produce the corresponding acid and amides from the product obtained with this preferred reagent.
Where necessary, hydroxy groups in intermediate compounds may be protected 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)]. A particularly useful protecting group which may be mentioned is tbutyldimethylsilyl.
Compounds in which R4 represents halogen and compounds in which R6 represents a leaving group are useful in the production of corresponding compounds of formula I.
The compounds of formula I may be isolated from their reaction mixtures using conventional techniques.
The compounds of formula I are useful because they possess pharmacological activity in animals; in particular they are 2q useful because they possess immunosuppressive activity, eg in the tests set out in Tests A, B, C and D. Thus the compounds, are indicated for use in the treatment or prevention of resistance to transplanted organs or tissues, such as kidney, heart, lung, bone marrow, skin, cornea, etc; and of autoimmune, inflammatory, proliferative and Μ O hyperproliferative diseases, and of cutaneous manifestations of immunologically-mediated diseases: for example rheumatoid arthritis, lupus erythematosus, systemic lupus erythematosus, Hashimoto's thyroiditis, multiple sclerosis, myasthenia gravis, . type 1 diabetes, uveitis, nephrotic syndrome, psoriasis, atopical dermatitis, contact dermatitis and further eczematous dermatitides, seborrheic dermatitis, Lichen planus, Pemphigus, bullous Pemphigoid, Epidermolysis bullosa, urticaria, angioedemas, vasculitides, erythemas, cutaneous eosinophilias, Alopecia areata, eosinophilic fasciitis, atherosclerosis etc. Ιθ The compounds of the invention are also indicated more generally in the treatment of respiratory diseases, for example reversible obstructive airways disease.
Further, the compounds of the invention are indicated in the treatment of a disease selected from intestinal inflammations/allergies such as Coeliac disease, proctitis, eosinophilic gastroenteritis, mastocytosis, Crohn's disease and ulcerative colitis; and food related allergic diseases which have symptomatic manifestation remote from the 2q gasto-intestinal tract, for example migraine, rhinitis and eczema.
The compounds of the invention are also indicated for use as antimicrobial agents, and thus may be used in the treatment of diseases caused by pathogenic microorganisms and the like.
We therefore provide the use of compounds of formula I as pharmaceuticals.
Further, we provide the use of a compound of formula I in the manufacture of a medicament for use as an immunosuppressive agent.
For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired IQ (eg topical, parenteral or oral) and the disease indicated. However, in general, satisfactory results are obtained when the compounds are administered at a daily dosage of from 0.001 to 20mg per kg of animal body weight.
For man the indicated total daily dosage is in the range of from O.Olmg to lOOOmg and preferably from 0.5mg to lOOmg, 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 2q administration, eg oesophageally, comprise from O.Olmg to 500mg, and preferably 0.5mg to lOOmg of the compound preferably admixed with a solid or liquid pharmaceutically acceptable diluent, carrier or adjuvant.
According to our invention we also provide a pharmaceutical composition comprising preferably less than 80%, and more preferably less than 50% by weight, of a compound of formula I 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 preferably is in a form having a 0.01 to ΙΟμιη. The suitable preserving, solubilisers (eg a such as hydroxypropyl mass median diameter of from IQ compositions may also contain stabilising and wetting agents, water-soluble cellulose polymer methylcellulose, or a water-soluble glycol such as propylene glycol), sweetening and colouring agents and flavourings. 15 The compositions may, if desired, be formulated in sustained release form.
For the treatment of reversible obstructive airways 2θ disease, we prefer the compound of formula I to be administered by inhalation to the lung, especially in the form of a powder.
According to a further aspect of the invention, there is 25 provided a method of effecting immunosuppression which comprises administering a therapeutically effective amount of a compound of formula I, as defined above, to a patient.
The compounds of formula I have the advantage that they are less toxic, more efficacious, are longer acting, have a broader range of activity, are more potent, are more stable, 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 have a number of chiral centres and may exist in a variety of stereoisomers. The invention provides all optical and stereoisomers, as well as racemic mixtures. The isomers may be resolved or separated by conventional techniques.
However, the preferred stereochemistry of various chiral carbon atoms are shown in formula Ia, Ilia and Y represents a cyclic group of formula Ila or Ilia, Test A Mixed Lymphocyte Reaction (MLR) I Ιθ The MLR test was performed in microtitre plates, with each well containing 5 χ 105 C57BL/6 responder cells (H-2b), χ 105 mitomycin C treated (25Mg/ml mitomycin C at 37°C for 30 minutes and washed three times with RPMI 1640 medium) BALB/C stimulator cells (H-2^) in 0.2ml RPMI 1640 15 medium supplemented with 10% fetal calf serum, 2mM sodium penicillin (50Mg/ml) and The cells were incubated at 37°C in a humidified atmosphere of 5% carbon dioxide and 95% of air for 68 hours and pulsed with 3H-thymidine 2Q (0.5μΰϊ) 4 hours before the cells were collected. The object compound of this invention was dissolved in ethanol and further diluted in RPMI 1640 medium and added to the cultures to give final concentrations of O.lMg/ml or less.
Test B hydrogen carbonate, streptomycin (50gg/ml).
Mixed Lymphocyte Reaction (MLR) II The MLR test was performed in 96-well microtitre plates with each well containing 3 x 10° cells from each of two - 23 responding donors in a final volume of 0.2ml RPMI 1640 medium supplemented with 10% .human serum, L-glutamine and penicillin/streptomycin. The compound under test was dissolved at lOmg/ml in ethanol and further diluted in RPMI 1640. The cells were incubated at 37°C in a humidified atmosphere at 5% carbon dioxide for 96 hours. 3H-thymidine (0.5gCi) was added for the final 24 hours of the incubation to provide a measure of proliferation.
Test C Ιθ Graft versus Host Assay (GVH) Spleen cells from DA and DAxLewis FI hybrid rats were prepared at approximately 108 cells/ml. 0.1ml of these suspensions were injected into the rear footpads of DAxLewis FI rats (left and right respectively). Recipient animals are dosed with the compound under test, either orally or subcutaneously, on days 0-4. The assay is terminated on day 7 when the popliteal lymph nodes of the animals are removed and weighed. The increase in weight of the left node relative to the weight of the right is a 2Q measure of the GVH response.
Test D Inhibition of Interleukin-2 (IL-21 secretion The test was performed following the method of S Sawada et al, J Immunol (6), Vol 139, ppl797-1803, but using the Jurkat cell line.
The invention is illustrated, but in no way limited, by the following Examples.
Example 1 17- Al1ν1-14-hydroxy-12-Γ 2-(4-hvdroxv-3-methoxvcvclohexvD-lmethvlvinvll-1.23.25-trimethoxv-13,19.21,27-tetramethvl11,28-dioxa-4-azatricvclo f 22.3.1.04·91octacos-18-ene5 2,3,10,16-tetraone (a) 17-Allvl-2,14-dihvdroxv-12-Γ 2-f 4-hvdroxv-3-methoxv cvclohexvl)-1-methvlvinyll-1.23,25-trimethoxy-13,19,21.27tetramethvl-11.28-dioxa-4-azatricvclo Γ 22.3.1.04·9loctacos18- ene-3.10.16-trione 17- Ally1-1,2,14-trihydroxy-12-[2-(4-hydroxy-3-methoxy cyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.04·9]octacos18- ene-3,10,16-trione (the compound of Example 5, WO 89/05304) (200mg) was added to a suspension of montmorillonite K10 (500mg) in methanol (5ml).
After stirring for 4 days at room temperature a further portion of montmorillonite was added (500mg) and stirring was continued for a further 2 days. The reaction mixture was then filtered through celite and was concentrated to an oil 2Q in vacuo. Column chromatography on silica then gave the subtitle compound as an oil (42mg).
MS: 843 [M+Na]+; 904 [M+Rb]+ (b) 17-Allyl-14-hvdroxv-12 - Γ 2 - (4-hvdroxy-3-methoxvcyc1o hexvll-1-methvlvinvl1-1.23,25-trimethoxy-13.19.21.27tetramethvl-11.28-dioxa-4-azatricvcloi22.3.1.04·9loctacos18-ene-2.3,10,16-tetraone The compound of step (a) (40mg) was dissolved in methanol (3ml) and to this was added cupric acetate (lOOmg). The resulting suspension was stirred and heated to reflux for 30 minutes. The reaction mixture was then cooled, filtered and evaporated in vacuo. Column chromatography on silica gave the title compound (30mg) as an oil.
MS (FAB): 902.5 [M+Rb]+; 840.8 [M+Na]+; 818.8 [M+H]+; 800.8 [M+H]+; 786.8 [M+H-CH3OH]+13C NMR 6: 211.7 (C16); 197.6 (C2); 169.3 (CIO); 166.2 (C3); 139.1 (C29); 130.5 (C31); 123.4 (C18); 116.7 (C42); 102.4 (Cl); 102.4 (Cl); 50.6 (C1-OCH3) IO Example 2 17-Allvl-l-amino-14-hvdroxv-12-Γ 2-(4-hvdroxv-3-methoxvcvclo hexyl)-1-methvlvinvll-23,25-dimethoxv-13,19,21,27-tetra methvl-11.28-dioxa-4-azatricvclor22.3.1.04·91octacos-18ene-2,3,10.16-tetraone and 17- Allvl-l.l4-dihvdroxv-12-Γ 2-(4-hvdroxv-3-methoxvcvclo hexvl)-1-methvlvinvll-23.25-dimethoxv-2-imino-13,19,21.27tetramethvl-11.28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos18- ene-3.10.16-trione 2Q (a) 17-Allvl-l-chloro-14-hydroxy-12-Γ2-(4-hvdroxv-3-methoxy cvclohexvl)-1-methvlvinvll-23.25-dimethoxv-13,19,21.27-tetra methvl-11.28-dioxa-4-azatricvclor 22.3.1.04'91octacos-18ene-2.3.10.16-tetraone A solution of 17-allyl-l,14-dihydroxy-12-[2-(4-hydroxy-32g methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo [22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (FR-900506) (0.6g) in dry dichloromethane (15ml) was added dropwise over 5 minutes at room temperature under an atmosphere of nitrogen to a solution of thionyl chloride (544μ1) and pyridine (1.33ml) in dry dichioromethane (15ml). After stirring for 5 minutes at room temperature the reaction mixture was added slowly to vigorously stirred saturated aqueous sodium hydrogen carbonate solution (50ml). After stirring for 5 minutes this mixture was extracted with diethyl ether (150ml) and the extract washed with dilute aqueous hydrochloric acid (1M, 50ml), water and IQ brine before being dried (MgSO4), filtered and evaporated in vacuo to give the subtitle compound as a foam (630mg).
MS: 908.4 [M+Rb]+; 906.4 [M+Rb] 870.7 [M-HCl+Rb] 844.9 [M+H]' 13, ’C NMR (CDC13) 6: 212.1 (C16); 189.3 (C2); 169.3 15 (CIO); 164.1 (C3); 140.4 (C19); 135.8 (C41); 132.3 (C29); 129.4 (C31); 122.6 (C18); 116.6 (C42); 108.9 (Cl); 84.3 (C34); 70.3 (C14); 48.2 (C20); 41.3 (C13); 9.8 (C39) (b) 17-Allvl-l-amino-14-hvdroxv-12-r2-(4-hvdroxv-3-methoxv cvclohexvl)-1-methvlvinyl1-23,25-dimethoxv-13,19,21,27-tetra methvl-11.28-dioxa-4-azatricvcloΓ 22.3.1.04'91octacos-18ene-2.3,10,16-tetraone and 17- Allvl-l,14-dihvdroxv-12-Γ 2-f 4-hvdroxv-3-methoxvcvclo hexyl)-1-methvlvinvll-23.25-dimethoxv-2-imino-13,19,21,2725 tetramethyl-11.28-dioxa-4-azatricyclor22.3.1.04'91octacos18- ene-3,10,16-trione A crude sample of the compound of step (a) (405mg) was taken up in THF (tetrahydrofuran) (8ml) and to this was added concentrated aqueous ammonia solution (4ml). After stirring for 20 minutes at room temperature the reaction mixture was diluted with water (20ml) and diethyl ether (50ml). The organic extract was then separated and washed 5 with brine before being dried (MgSO4), filtered and concentrated in vacuo to a foam. This was chromatographed on silica using HPLC eluting with 2% methanol in diethyl ether to give fraction A (190mg) and fraction B (98mg). Fraction A was further purified by chromatography on silica 1θ using HPLC eluting with ethyl acetate to give the first title compound (92mg) as a foam.
MS: 887.5 [M+Rb]+; 803.7 [M+H]+13C NMR (CDC13) 5: 213 (C16); 198.2 (C2); 169.2 (CIO); 166.2 (C3) ; 139.4 (C19); 135.7 (C41); 132.6 (C29); 129.615 (C31) ; 122.2 (C18); 116.5 (C42); 88.6 (Cl); 84.2 (C34); 76.7 (C12) ; 75.5 (C23); 71.1 (C24) ; 70.2 (C14); 56.4 (C9); 52.7 (C17); 48.6 (C20); 43.0 (C15) ; 39.9 (C13); 38.9 (C5); 31.3 (C36) ; 30.7 (C37); 27.9 (C8) ; 26.1 (C21); 24.6 (C6); 21.3 (C7); 20.4 (C44); 14.2 (C30); 9.5 (C39) 2Q Fraction B was further purified by chromatography on silica using HPLC eluting with hexane/acetone [2:1] to give the second title compound (70mg) as a foam.
MS: 887.5 [M+Rb]+; 825.7 [M+Na]+; 803.7 [M+H]+; 785.7 [M+H-H2O]+; 767.7 [M+H-2H2O]+2513C NMR (CDC13) 6: 214.4 (C16); 175.7 (C2); 169.9 (CIO); 168 (C3); 139.1 (C19); 134.7 (C41); 131.3 (C29); 128.2 (C31); 123.4 (C18); 116.7 (C42); 95.5 (Cl); 84.2 (C34); 75.2 (C23); 73.4 (C25); 71.5 (C24); 69.5 (C14); 52.9 (C17); 49.8 (C20); 44.9 (C15); 39.6 (C13); 39.3 (C5) ; 31.2 (C36) ; 30.8 (C37) ; 27.7 (C8) 26.2 (C21) ; 24.3 (C6) ; 21.0 (C44); 20.0 (C7) ,-14.5 (C30) ; 10.2 (C39) Example 3 17-Allvl-l-(1-thiopropvl)-14-hvdroxv-12-Γ2-(4-hvdroxv-3methoxvcvclohexvl)-1-methvlvinvll-23.25-dimethoxv13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-2.3,10.16-tetraone A solution of the compound of Example 2(a) (lOOmg) and IQ propanethiol (0.1ml) in THF (2ml) and saturated aqueous sodium hydrogen carbonate solution (2ml) was stirred vigorously for 24 hours at room temperature. Water (10ml) was then added and the reaction mixture was extracted with diethyl ether (20ml). The organic extract was then washed with brine before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/acetone [3:1] then gave the title compound (42mg) as a foam.
MS: 946 [M+Rb]+; 885 [M+Na]+; 863 [M+H]+; 787 [M+H-CH3(CH2)2SH]+; 769 [M+H-CH3(CH2)2SH-H2O]+13C NMR (CDC13) δ: 212.8 (C16); 191 (C2)? 169.3 (CIO); 166.7 (C3); 140.8 (C19); 135.2 (C41); 131.3 (C29)i 128.7 (C31); 122.3 (C18); 116.8 (C42); 89.6 (Cl); 84.1 (C34); 73’9 (C25); 73.5 (C35); 70.2 48.9 (C20); 44.9 (C15); 39.4 33.3 (C26); 31.1 (C36); 30.7 27.4 (sch2) ; 24.4 (C6); 14.3 (C30); 13.6 (S(CH2)2CH3); 10.2 (C39) Example 4 17-Allvl-l-(N-acetvl)amino-14-hvdroxv-12-Γ 2-(4-hvdroxv-3methoxvcvclohexvl)-1-methvlvinvll-23,25-dimethoxv13.19.21,27-tetramethvl-ll.28-dioxa-4-azatricvclo 122.3.1.04'91octacos-18-ene-2.3.10.16-tetraone A sample of the first title compound of Example 2 (crude, lOOmg) was taken up in methanol (10ml) and acetic anhydride (0.6ml) was added. After being stored at 4°C for 3 days further acetic anhydride (0.3ml) was added and the reaction mixture was stored at this temperature for a further 2 days. The reaction mixture was then poured into saturated aqueous sodium hydrogen carbonate solution (100ml) and this was then extracted with diethyl ether (100ml). The separated organic extract after washing with brine was dried (MgSO4), filtered and concentrated in vacuo to a foam. Chromatography on silica eluting with dichloromethane/acetone in an increasing acetone gradient then gave material which was further purified by chromatography on silica eluting with ethyl acetate to give the title compound (37mg) as a foam.
MS: 929.1 [M+Rb] + ; 867.9 [M+Na] + ,° 846 [M+H] + ; 769.1 [M+H-H2O-CH3CONH2]+13C NMR (CDC13) 5: 212 (C16); 190.2 (C2) ; 169.8 (CIO); 169.4 (CH3CONH); 163.1 (C3); 140.2 (C19); 135.6 (C41); 132.2 (C29) ; 129.4 (C31) ; 122.2 (C18); 116.4 (C42) ; 87.8 (Cl) ; 84.2 (C34); 76.8 (C12); 76.3 (C23); 74.9 (C24); 70.4 (C14) ; 52.7 (C17); 51.2 (C9); 47.8 (C20); 45.1 (C15) ; 44.1 (C5) ; 41.6 (C13); 31.3 (C36); 30.6 (C37) ; 27.3 (C8); 26.0 (C21); 24.3 (C6); 22.9 (CH3CONH); 21.4 (C7); 18.3 (C44); 16.9 (C47); 15.5 (C43); 14.8 (C30); 9.5 (C39) Further elution then gave the Cl isomeric compound (46mg). MS: 929.1 [M+Rb]+; 867.5 [M+Na]+; 845.6 [M+H]+; 827.6 [M+H-0H]+; 768.6 [M+H-H2O-CH3CONH2]+13C NMR (CDC13) 6: 210.4 (C16); 194.3 (C2); 169.4 (CIO); 169.0 (CH3CONH); 166.1 (C3); 137.8 (C19); 135.7 1Q (C41); 131.7 (C29); 129.5 (C31); 123.7 (C18); 116.5 (C42); 89.7 (Cl); 84.2 (C34); 77.9 (C12); 76.0 (C24); 74.5 (C23); 69.8 (C14); 39.5 (C13); 28.2 (C21); 27.3 (C8); 25.2 (C6); 23.1 (CH3CONH); 21.5 (C7); 16.9 (C47); 13.2 (C30); 9.9 (C39) Example 5 17-Allvl-l-(N-formvl)amino-14-hvdroxv-12-Γ 2-(4-hvdroxv-3methoxvcvclohexvl)-1-methylvinyl1-23, 25-dimethoxv13.19,21.27-tetramethvl-ll.28-dioxa-4—azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3.10.16-tetraone 2q a) 17-Allvl-14-tbutvldimethvlsilvloxv-12-r2-(4^butvldimethvlsilvloxv-3-methoxvcvclohexvl)-1-methvlvinvl1 -23,25-dimethoxv-l-hvdroxv-13.19,21.27-tetramethvl-ll,28dioxa-4-azatricyclo Γ 22.3.1.04·91octacos-18-ene-2,3,10,16tetraone 25 To a solution of FR-900506 (500mg, 0.622mmole) in dry dichloromethane (20ml) at room temperature under nitrogen was added 2,6-dimethylpyridine (0.4ml) and tbutyldimethylsilyl triflate (362mg, 1.32mmole). After 30 minutes at room temperature further ^butyldimethylsilyl triflate (362mg, 1.32mmole) was added and the reaction mixture was stirred for a further 30 minutes at room temperature. Dichloromethane (30ml) was then added and the reaction mixture was extracted with dilute aqueous hydrochloric acid (25ml) and brine (25ml). The organic extract was dried (MgSO4), filtered and evaporated to an oil in vacuo. Purification by column chromatography on silica eluting with hexane/acetone [9:1] gave the title IQ compound (606mg, 94%) as an oil.
MS: 1055 [M+Na]+; 1117 [M+Rb]+ b) 17-Allyl-l-chloro-12-Γ 2-(4-tbutvldimethvlsilvloxv-3methoxvcvclohexvl)-1-methvlvinvl1-14-^butvldimethvl silvloxv-23.25-dimethoxv-13.19.21.27-tetramethvl-ll,28-dioxa -4-azatricvcloΓ22.3.1.04'91octacos-18-ene-2,3,10,16tetraone A sample of the compound of step (a) (lg) in dry dichloromethane (10ml) was added dropwise over 5 minutes to a stirred solution of thionyl chloride (0.35ml) and pyridine (0.94ml) in dry dichloromethane (10ml). After stirring for a further 5 minutes at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution (50ml) and this was extracted with diethyl ether (100ml). The separated organic extract after washing with dilute aqueous hydrochloric acid (1M, 50ml), water and brine was then dried (MgSO4), filtered and concentrated in vacuo to give the subtitle compound as a foam (lg). c) 17-Allvl-l-amino-12-r2-f4-tbutvldimethvlsilvloxv-3methoxvcvclohexvl)-1-methvlvinvl1-14-^butvldimethvl silvloxv-23.25-dimethoxv-13,19.21.27-tetramethvl-ll.28-dioxa -4-azatricvclo Γ 22.3.1.04·91octacos-18-ene-2.3.10.165 tetraone A sample of the crude subtitle compound from step (b) (744mg) was dissolved in THF (10ml) and this was then added dropwise to concentrated agueous ammonia solution (5ml). The reaction mixture after being stirred vigorously for 15 IQ minutes was diluted with water (25ml) and diethyl ether (50ml). The diethyl ether extract was then separated and was washed with brine before being dried (MgSO4), filtered and concentrated in vacuo to a foam. Chromatography on silica eluting with hexane/ethyl acetate [5:1] then gave the subtitle compound as a foam (250mg). d) 17-Allvl-l- (N-formvl) amino-12- Γ 2- (4-^^ν1ά^6ίΗν1 silyloxy-3-methoxycyclohexyl)-l-methylvinvll-14tbutvldimethvlsilvloxv-23.25-dimethoxv-13,19.21,27tetramethvl-ll ,28-dioxa-4-azatricvclo Γ22.3.1.04'9]octacos20 18-ene-2.3.10.16-tetraone To a crude sample of the subtitle compound from step (c) (120mg) in formic acid (4ml) at room temperature was added acetic anhydride (0.2ml). After stirring for 4 hours at room temperature the reaction was stored at 4’C for 16 hours before being poured into saturated agueous sodium hydrogen carbonate solution (100ml). After stirring this mixture for 20 minutes at room temperature it was extracted with diethyl ether (50ml) and this extract was then washed with brine before being dried (MgSO4), filtered and concentrated in vacuo to give, the subtitle compound as an oil. e) 17-Allvl-l-(N-formvl)amino-14-hvdroxv-12-Γ 2-f 4-hvdroxv5 3-methoxvcvclohexvl)-1-methvlvinvl1-23.25-dimethoxv13.19,21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'9]octacos-18-ene-2,3,10.16-tetraone A crude sample of the subtitle compound from step (d) (120mg) was taken up in methanol (3ml) and aqueous IQ hydrofluoric acid was added (0.2ml). After 2 hours at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution (20ml) and this was then extracted with diethyl ether (40ml). The separated organic extract was then washed with brine before being dried (MgSO4), filtered and concentrated in vacuo to a foam. Chromatography on silica eluting with hexane/acetone [2:1] then gave the title compound (30mg) as a foam.
MS: 915.2 [M+Rb]+; 831.6 [M+H] 2θ 768.6 [M+H-H2O-H2NCHO]+ 813.6 [M+H-H2O]+; C NMR (CDC13) S: 214.6 (C16); 193.6 (C2); 169.2 (CIO); 166.1 (C3); 159.9 (NHCOH); 137.2 (C19); 135.3 (C41); 122.6 (C18); 116.5 (C42); 88.7 (Cl); 84 (C34); 77.9 (C12); 69.2 (C14); 56.2 (C9); 48.7 (C20); 43.6 (C15); 39.9 (C13); 24.2 (C6); 20.8 (C44); 16.7 (C47); 14.7 (C43); 14.0 (C30); 11.1 (C39) Example 6 17-Al1vl-1-fluorο-14-hvdroxv-12-Γ 2-f 4-hvdroxv-3-methoxvcvc1o hexvl)-l-methvlvinvll-23,25-dimethoxv-13,19,21.27tetramethvl-ll .28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos18-ene-2,3,10.16-tetraone To a cold (0°C) solution of the subtitle compound of g Example 5(a) (250mg) in dry dichloromethane (10ml) under nitrogen was added diethylaminosulphur trifluoride (lOOmg). After stirring for 2 hours at 0°C the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution (30ml) and this was then extracted with IQ diethyl ether (100ml) washing with brine The separated organic extract after was dried (MgSO4), filtered and concentrated in vacuo to a foam (248mg). This was then dissolved in acetonitrile (10ml) and 40% aqueous hydrofluoric acid (0.2ml) was added. After being stirred 15 for two hours at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution (50ml) and this was then extracted with diethyl ether (100ml). The separated organic extract was then washed with brine and was dried (MgSO4), filtered and concentrated in vacuo to an oil. Chromatography on silica eluting with dichloromethane/acetonitrile [2:1] then gave the title compound (28mg) as a foam.
MS: 890.5 [M+Rb]+; 828.9 [M+Na]+; 787 [M+H-HF]+; 769 [M+H-HF-H2O]+19F NMR 5: -139.55 (d,J=28.15Hz); -141.55 (d,J=28.15Hz) (two rotamers) 13C NMR (CDC13) 6: 211.9 (C16); 192.3 (C2); 169 (CIO); 164.2 (C3); 140 (C19)? 135.6 (C41); 132 (C29) ; 129.5 (C31); 122.8 (C18) ; 116.5 (C42); 112.8 (Cl); 84.2 (C34); 77.2 (C12) ; 76.0 (C23); 75.1 (C25); 73.5 (C35); 72.5 (C24); 69.8 (C14) ; 48.1 (C20)? 45 (C5); 43.8 (C15); 40.8 (C13); 32.3 (C26) ; 31.2 (C36); 30.7 (C37); 26.8 (C8); 25.9 (C21); 25.0 (C6) ; 21.7 (C7); 19.4 (C44); 15.8 (C47); 15.1 (C43); 14.5 (C30); 9.7 (C39) Example 7 The first title compound of Example 2 was tested in Test D, and found to inhibit IL-2 secretion by 50% (IC50) at a IQ concentration of 2xlO”loM.
Example 8 17-Allvl-l,14-dihvdroxv-12-Γ2-(cvclopentyl-3-carboxaldehvde) -1-methvlvinvll-23,25-dimethoxv-13,19.21,27-tetramethvl11.28-dioxa-4-azatricvclo r 22.3.1.04·91octacos-18-ene15 2,3.10.16-tetraone (a) 17-Allyl-1-hvdroxv-12-Γ 2-f 4-hvdroxv-3-methoxvcvc1o hexvl)-1-methvlvinvl1-14-tbutvldimethvlsilvloxv-23,25dimethoxv-13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3,10,16-tetraone 2Q A solution of the product from Example 5(a) (1.28g) in methanol (100ml) containing pyridinium p-toluene sulphonate was stirred for 18 hours at room temperature. Volatiles were then removed in vacuo and the residue was dissolved in diethyl ether. The ethereal solution after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine was dried (MgSO4), filtered and evaporated in vacuo to give the subtitle compound as a pale yellow foam (0.97g). (b) 17-Allyl-l-hvdroxv-12-r2-f4-trifluoromethvlsulphonvloxv -3-methoxvcvclohexyl)-1-methvlvinvlΊ-M-^butvldimethvl silvloxv-23.25-dimethoxv-13.19.21,27-tetramethyl-11.28-dioxa -4-azatricvcloΓ22.3.1.04'91octacos-18-ene-2,3.10.16tetraone To a cold (-10°C) stirred solution of the product of step (a) (0.97g) in dry dichloromethane (25ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.1ml). Ιθ After stirring for 15 minutes at -10’C, saturated aqueous sodium hydrogen carbonate solution was added and the reaction mixture was extracted with diethyl ether. The ether extracts were then washed with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous 15 hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine before being dried (MgSO4), filtered and concentrated in vacuo to give the title compound as an oil (0.95g). (c) 17-Allyl-l-hvdroxv-12-Γ 2-(cvclopentyl-3-carboxaldehvde) 2q -1-methvlvinvl1-14-tbutvldimethvlsilvloxv-23,25-dimethoxv13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene-2.3.10.16-tetraone Silica (55g, Merck Kieselgel 60) was added to a solution of the product from step (b) (0.9g) in dichloromethane 25 (250ml). Volatiles were then removed in vacuo at room temperature and the resulting freely flowing powder was stored at 8°C for 16 hours. The support was then washed with ethyl acetate and 10% acetone in ethyl acetate containing 2,6-dimethylpyridine. The combined organic extracts after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate 5 solution and brine were dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an acetone gradient then gave the title compound (0.126g) as a foam. (d) 17-Allvl-l,14-dihvdroxv-12-r2-fcvclopentvl-3carboxaldehvde)-1-methvlvinvll-23,25-dimethoxv-13.19.21,27tetramethvl-ll .28-dioxa-4-azatricvclo Γ 22.3.1.04·9 1 octacos18-ene-2,3.10.16-tetraone To a solution of the compound of step (c) (25mg) in acetonitrile (5ml) was added 40% aqueous hydrofluoric acid 15 (1ml). After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and the mixture was extracted with diethyl ether. The organic extract was then dried, (MgSO4), filtered and evaporated to an oil in vacuo. 2q Chromatography on silica eluting with acetone/hexane [1:2] then gave the title compound (18mg) as a foam.
Example 9 1,14-dihvdroxv-i2-r2-(cvclopentvl-3-carboxaldehvde)-1methvlvinvll-23.25-dimethoxv-17-propvl-13.19,21,272g tetramethyl-11,28-dioxa-4-azatricvclor22.3.1.04·9]octacos18-ene-2,3.10.16-tetraone To a solution of the product of Example 8 (15mg) in methanol (4ml) was added Pd-on-C (4mg, 10%) and the resulting suspension was then stirred in an atmosphere of hydrogen for 1 hour at O’C. The reaction mixture was then filtered and volatiles were removed in vacuo. Chromatography on silica then gave the title compound as a foam (13mg).
Example 10 17-Allvl-l.14-dihvdroxv-12-Γ2-(cvclopentvl-3-methanol)1-methvlvinvl1-23,25-dimethoxv-13,19,21.27-tetramethvl10 ll ,28-dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene2.3.10.16-tetraone (a) 17-Allvl-l-hvdroxv-12-Γ2-(cvclopentvl-3-methanol)-1methvlvinvl1-14-tbutvldimethvlsilvloxv-23,25-dimethoxv13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-2.3.10.16-tetraone To a solution of the product of Example 8(c) (170mg) in dry THF (15ml) at -70’C was added a solution of L-selectride in THF (1M) slowly under nitrogen until no starting material remained (0.4ml). Saturated agueous ammonium chloride solution (0.5ml) was then added at -70 °C followed by aqueous hydrogen peroxide solution (30% by weight, 1ml) and ethanolamine (0.1ml). After warming to O’C the reaction mixture was extracted with diethyl ether and this was washed with water (x2), dilute aqueous hydrochloric acid saturated aqueous sodium hydrogen carbonate before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with acetone/hexane [2:7] then gave the title compound (151mg) as a foam.
(IN) and solution, MS (FAB): 911 [M+Na]+; 972 [M+Rb]+. (b) 17-Allyl-l.14-dihvdroxv-12-Γ2-(cyclopentyl-3-methanol)1-methvlvinvl1-23,25-dimethoxv-13.19,21.27-tetramethvlll . 28-dioxa-4-azatricyclo Γ 22.3.1.octacos-18-ene5 2,3,10.16-tetraone To a solution of the product of step (a) (150mg) in acetonitrile (20ml) was added 40% aqueous hydrofluoric acid (3ml)' After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium IQ hydrogen carbonate solution and the mixture was extracted with diethyl ether. The organic extracts were then dried, (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with acetone/hexane [1:3] then gave the title compound (130mg) as a foam.
MS (plasma spray): 738.54 [M+H-2H2O] 756.58 [M+H-H2O]+; 774.6 [M+H]+; 791.57 [M+NH4]+ 13, ’C NMR (CDC13) 6: (Major rotamer) 212.5 (C16); 196.2 (C2); 169 (CIO); 164.7 (C3); 138.8 (C19); 135.5 (C40); 131.4 (C31); 131 (C29)} 122.4 (C18); 116.5 (C41); 97 2q (Cl); 77.7 (C12); 75 (C23); 69.9 (C14); 67 (C37); 56.5 (C9); 48.5 (C20); 43.6 (C15); 27.6 (C8); 26 (C21); 24.4 (C6); 20.9 (C7); 20.3 (C43); 13.9 (C30); 9.5 (C38) Example 11 1,14-dihvdroxv-12-Γ 2-(cvclopentvl-3-methanol)-1-methvlvinvl1 25 -23.25-dimethoxv-17-propvl-13,19,21.27-tetramethvl-ll.28dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene-2,3,10,16tetraone To a solution of the title compound of Example 10 (22mgj in methanol (10ml) was added 10% Pd-on-C (5mg) and the resulting suspension was then stirred in an atmosphere of hydrogen for 2 hours at O’C. The reaction mixture was then filtered and volatiles were removed in vacuo.
Chromatography on silica then gave the title compound as a foam (18mg).
MS (plasma spray): 794 [M+NH4]+ Example 12 17-Allvl-l-hvdroxv-12-r2-(cvclopentvl-3-carboxylic acid) 1Q -1-methvlvinvll-14-tbutvldimethvlsilvloxv-23,25-dimethoxv13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricyclo Γ22.3.1.04·9 ) octacos-18-ene-2.3,10,16-tetraone To a solution of the product of Example 8(c) (393mg) in tbutanol (30ml) containing 1-methylcyclohex-l-ene (4ml) was added dropwise a solution of sodium chlorite (0.75g) and sodium phosphate (0.75g) in distilled water (10ml). After stirring for 10 minutes at room temperature the reaction mixture was partitioned between ethyl acetate and water and the organic extract was separated. This was then 2q washed with aqueous sodium phosphate solution, an aqueous sodium thiosulphate/sodium phosphate mixture and aqueous sodium phosphate solution before being dried (MgSO4), filtered and evaporated in vacuo to give the title compound (350mg) as a foam.
Example 13 17-Al1vl-1.14-dihvdroxv-12-(2-(cvc1opentvl-3-carboxylic acid)-1-methvlvinvll-23,25-dimethoxv-13.19,21,27tetramethvl-ll . 28-dioxa-4-azatricvclo Γ 22.3.1.octacos41 18-ene-2.3.10.16-tetraone To a solution of the product of Example 12 (350mg) in acetonitrile (30ml) was added 40% aqueous hydrofluoric acid (3ml). After stirring for 1.5 hours at room temperature the reaction mixture was poured into ethyl acetate and the organic extract was washed with water and saturated aqueous sodium phosphate solution (x4) before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with acetone/hexane/acetic Ιθ acid [40:10:1] then gave the title compound (32mg) as a foam.
MS (FAB): 771.02 [M-OH+H]+; 811 [M+Na]+; 872.72 [M+Rb]+13C NMR 6: (Major rotamer) 212.6 (C16); 196.1 (C2) ; 181.6 (C37); 169.1 (CIO); 164.7 (C3); 138.9 (C19); 135.6 (C40); 132.7 (C29); 130.3 (C31); 122.6 (C18); 116.7 (C41); 98.6 (Cl); 77.8 (C12); 75.3 (C23); 73.6 (C25); 72.6 (C24); 70.0 (C14); 56.7 (C9); 52.9 (C17); 48.7 (C20); 26.3 (C21); 24.6 (C6); 21.1 (C7); 20.4 (C43); 14.1 (C30); 9.7 (C38). 2q Example 14 17-Allvl-l.14—dihvdroxv-12-Γ 2-(cvclopentvl-3-carboxvlic acid methyl ester)-1-methvlvinvl1-23.25-dimethoxv13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo >4.9 Γ 22.3.1. O*·*·2!octacos-18-ene-2,3,10,16-tetraone To a solution of the product of Example 13 (25mg) in diethyl ether (5ml) at 0°C was added diazomethane.
Volatiles were then removed in vacuo to give the title compound as a foam (25mg).
Example 15 1>14-dihvdroxv-12-r2-(cvclopentvl-3-carboxvlic acid methyl ester)-1-methvlvinvl1-23,25-dimethoxy-17-propyl-13.19.21,27tetramethvl-ll, 28-dioxa-4-azatricvclor 22.3. 1.0^-«-^loctacos5 18-ene-2.3.10.16-tetraone To a solution of the product of Example 14 (20mg) in methanol (10ml) was added 10% Pd-on-C (4mg) and the resulting suspension was then stirred in an atmosphere of hydrogen for 2 hours at O’C. The reaction mixture was then IQ filtered and volatiles were removed in vacuo. Chromatography on silica then gave the title compound as a foam (17mg).
Example 16 1.14-dihvdroxv-12-r2-(cvclopentvl-3-carboxvlic acid) -1-methvlvinvl1-23,25-dimethoxv-17-propvl-13.19.21,27tetramethvl-ll, 28-dioxa-4-azatricvclor22.3. 1.0-^-^loctacos18-ene-2.3.10,16-tetraone To a solution of the product of Example 15 (18mg) in methanol (10ml) was added 10% Pd-on-C (4mg) and the 2Q resulting suspension was then stirred in an atmosphere of hydrogen for 2 hours at O’C. The reaction mixture was then filtered and volatiles were removed in vacuo. Chromatography on silica then gave the title compound as a foam (17mg).
MS (FAB): 874 [M+Rb]+ Example 17 17-Allvl-l-hvdroxv-12-Γ 2-f cvclopentvl-3-methyl propenoate)-1-methvlvinvl1-14-^butvldimethvlsilvloxv43 23.25-dimethoxv-13,19.21.27-tetramethvl-ll.28-dioxa-4azatricvclo Γ 22♦3♦1.04'91octacos-18-ene-2.3.10.16-tetraone A solution of the product of Example 8(d) (140mg) and (carbomethoxymethylene)triphenylphosphorane (140mg) in dry distilled toluene (10ml) was stirred and heated at 70°C for one hour. After stirring at room temperature overnight the reaction mixture was diluted with diethyl ether and this was then washed with saturated aqueous sodium hydrogen carbonate solution and brine. The organic extract was then IQ dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing diethyl ether gradient then gave the title compound (70mg) as a foam.
Example 18 17-Allvl-l.14-dihvdroxv-12-r2-fcvclopentvl-3-methvl propenoate)-l-methvlvinvll-23.25-dimethoxv-13,19,21,27tetramethvl-ll . 28-dioxa-4-azatricvclor22.3.1.04 ·9J. octacos-18-ene-2,3.10.16-tetraone To a solution of the product of Example 17 (70mg) in 2q acetonitrile (10ml) was added 40% aqueous hydrofluoric acid (lml). After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and the mixture was extracted with diethyl ether. The combined ether extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, were dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound (55mg) as a foam.
MS (plasma spray): 792.78 [M+H-2H2O]+; 810.80 [M+H-H2O]+; 828.86 [M+H]+; 845.84 [M+NH4]+ MS (negative plasma spray): 826.09 [M-H]+5XH NMR (CDC13) Si 6.93 (1H, dd, J=8.1 and 16.6 Hz); .78 (1H, d, J=5.78 Hz), 3.71 (3H, s, CO2Me) NMR : (Major rotamer) 212.4 (C16); 196.1 (C2); 153.3 (C38); 138.8 (C19) ; 135.4 (C43); 122.6 (C18) ; 119 (C37) ; 116.6 (C44); 97.1 (Cl); 56.6 (C9); 51.3 (C40); 9.7 10 (C41).
Example 19 1«14-dihvdroxv-12-r2-fcvclopentvl-3-carboxaldehvde)-1-methvl vinvn-23,25-dimethoxv-17-ethvl-13.19.21.27-tetramethvlll .28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene15 2,3,10.16-tetraone (a) 1-Hvdroxv-12-Γ2-fcvclopentvl-3-carboxaldehvde) -1-methv1 vinvll-14-^'butvldimethvlsilvloxv-23,25-dimethoxv-17-ethvl13,19.21.27-tetramethvl-ll,28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-l8-ene-2.3.10.16-tetraone 2θ The subtitle compound was prepared from FR-900520 in a manner analogous to the compound of Example 8(c). (b) 1.14-dihvdroxv-12-r2-(cvclopentvl-3-carboxaldehvde)-1methvlvinvll-23.25-dimethoxv-17-ethvl-13,19,21« 27tetramethvl-ll .28-dioxa-4-azatricvclo Γ 22.3.1.04'91 octacos25 18-ene-2.3,10.16-tetraone The product of step (a) was deprotected following the method of Example 8(d) to give the title compound.
Example 20 1.14- dihvdroxv-12-r2-(cvclopentyl-3-methanol)-1-methvlvinvl1 -23,25-dimethoxv-17-ethvl-13,19,21.27-tetramethvl-ll,28dioxa-4-azatricvclor22.3.1.04·91octacos-18-ene-2.3,10.16tetraone The product of Example 19 was reduced by the method of Example 10(a) to give the title compound.
MS (plasma spray): 779 [M+NH4]+ Example 21 1.14- dihvdroxv-12-r2-icvclopentvl-3-carboxvlic acid) Ιθ -1-methvlvinvl1-23.25-dimethoxv-17-ethyl-13.19,21.27tetramethvl-ll , 28-dioxa-4-azatricvclo Γ22.3.1.04,91octacos18-ene-2,3,10,16-tetraone Oxidation of the product of Example 19(a) following the method of Example 12 and then deprotection following the method of Example 13 gave the title compound.
MS (FAB): 709 [M+Na]+ Example 22 1.14- dihvdroxv-12-r2-(cvclopentyl-3-carboxvlic acid methyl ester)-1-methylvinyl)-23,25-dimethoxv-17-ethvl-13,19,21,272q tetramethvl-11.28-dioxa-4-azatricvcloΓ22.3.1.04·9loctacos18-ene-2.3.10,16-tetraone Esterification of the product of Example 21 following the method of Example 14 yielded the title compound.
Example 23 1.14-dihvdroxv-12-r2-(cvclopentvl-3-methvl propenoate)-1methvlvinvll-23.25-dimethoxv-17-ethvl-13.19.21.27-tetra methvl-11.28-dioxa-4-azatricvclo Γ 22.3.1.04.91octacos-18-ene2.3.10.16-tetraone Wittig reaction on the product of Example 19(a) following the method of Example 17 and then deprotection following the method of Example 18 gave the title compound.
MS (plasma spray): 834 [M+NH4]+ Example 24 l-Hvdroxv-12-r2-(cvclopentvl-3-carboxaldehyde)-1methvlvinyl1-23,25-dimethoxv-17-propvl-13,19,21,27tetramethvl-ll ,28-dioxa-4-azatricvclo Γ22.3.1.04'9!octacos18-ene-2.3.10.16-tetraone a) l-Hvdroxv-12-r2-(4-trifluoromethvlsulphonyloxv-3methoxycyclohexyl)-l-methvlvinvll-23,25-dimethoxv-17-propvl13,19.21.27-tetramethvl-ll,28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-2.3,10,16-tetraone To a cold (-10’C), stirred solution of l-hydroxy-12-[2-(425 hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy17-propyl-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (Example 12, WO 89/05304) (0.3g) in dry dichloromethane (12ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.1ml) until no starting material remained. Saturated aqueous sodium hydrogen carbonate solution was then added and the reaction mixture was extracted with diethyl ether. The ether extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), and saturated aqueous sodium hydrogen carbonate solution, were dried (MgSO4), filtered and concentrated in vacuo to give the title compound as an oil (300mg). b) l-Hvdroxv-12-Γ2-(cvclopentvl-3-carboxaldehvde)-1methvlvinyl1-2 3.2 5-dimethoxv-17-propyl-13,19,21,27tetramethvl-ll .28-dioxa-4-azatricvclo Γ22.3.1.04,91octacos18-ene-2.3.10.16-tetraone 5 Silica (18g, Merck Kieselgel 60) was added to a solution of the product of step (a) (300mg) in dichloromethane (100ml). Volatiles were then removed in vacuo at room temperature and the resulting freely flowing powder was stored at 8'C for 16 hours. The support was then washed with acetone containing triethylamine and the solvent was evaporated in vacuo to an oil. Chromatography on silica eluting with hexane in an acetone gradient then gave the title compound as a foam (51mg).
Example 25 l-Hvdroxv-12-Γ2-(cvclopentvl-3-methanol)-1-methvlvinvl123°, 25-dimethoxv-17-propyl-13,19,21,27-tetramethyl-11.28dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene-2,3,10,16tetraone Reduction of the product of Example 24 following the method 2q of Example 10(a) yielded the title compound.
Example 26 l-Hvdroxv-12-Γ 2-(cyclopentyl-3-carboxvlic acid)-1-methyl vinvll-23.25-dimethoxv-17-propvl-13.19.21.27-tetramethvlll .28-dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene25 2,3,10,16-tetraone Oxidation of the product of Example 24 using the method of Example 12 gave the title compound.
Example 27 l-Hvdroxv-12-r2-(cvclopentvl-3-carboxvlic acid methvlester) -1-methvlvinvl1-23,25-dimethoxv-17-propvl-13,19,21,27-tetra methvl-11,28-dioxa-4-azatricvclo Γ 22.3♦1.04'91octacos-18ene-2,3.10.16-tetraone Esterification of the product of Example 18 using diazomethane following the method of Example 14 gave the title compound.
Example 28 l-Hvdroxv-12-r2-(cvclopentvl-3-methyl propenoate)-1-methvl vinyl]-23.25-dimethoxv-17-propyl-13.19,21.27-tetramethvlll ,28-dioxa-4-azatricvclor 22.3.1.04·91octacos-18-ene2.3.10.16- tetraone Wittig reaction with the product of Example 24 following the method of Example 17 yielded the title compound.
Example 29 l-Hvdroxv-12-Γ 2-(cvclopentvl-3-carboxaldehyde)-1-methvl vinyl1-23,25-dimethoxv-17-ethyl-13,19,21.27-tetramethvlll ,28-dioxa-4-azatricvcio Γ 22.3.1.04'91octacos-18-ene2.3.10.16- tetraone 2q (a) l-Hvdroxv-12-r2-f4-hvdroxv-3-methoxvcvclohexvl)-1methvlvinvll-23.25-dimethoxv-17-ethvl-13.19.21.27-tetra methvl-11,28-dioxa-4-azatricvclo Γ22.3.1.04'9]octacosa14.18-ene-2.3.10.16-tetraone 1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyi)-1methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28dioxa-4-azatricyclo[22.3.1.04·9]octacos-18-ene-2,3,10,16tetraone (FR-900520) (lOOmg) and p-toluenesulphonic acid (2mg) were dissolved in dry toluene (20ml) and were heated for 2 hours at 100°C under an atmosphere of nitrogen. Removal of solvent in vacuo and chromatography on silica eluting with hexane/acetone [2:1] gave the sub-title compound as a foam (80mg). MS (FAB): 774.8 [M+H]+; 796.85 [M+Na]+; 858.71 [M+Rb]+.13C NMR <5: (major rotamer) 201.15 (C16); 196.0 (C2); 169.2 (CIO); 165.1 (C3); 147 .8 (C15); 138.0 (C19); 123.82 (C18); 97.88 (Cl); 84. 05 (C34). (b) l-Hvdroxv-12-r2-(4-hvdroxv-3-methoxvcvclohexvl)-1methvlvinvl1-23,25-dimethoxv-17-ethvl-13.19.21.27-tetra methvl-11,28-dioxa-4-azatricvclo Γ 22.3.1.04·9loctacos-18ene-2,3,10,16-tetraone A sample of the product from step (a) was dissolved in 15 methanol (20ml) and 10% Pd-on-carbon (lOmg) was added. The mixture was stirred in an atmosphere of hydrogen for 1.5 hours at room temperature and pressure, and was then filtered through celite and evaporated to an oil in vacuo♦ Column chromatography on silica eluting with hexane/acetone 20 [2:1] gave the subtitle compound as a foam (50mg).
MS (FAB): 776 [M+H]+; 798 [M+Na]+; 860 [M+Rb]+. 13C NMR 6: (major rotamer) 212.34 (C16); 196.42 (C2); 169.38 (CIO); 165.16 (C3); 138.9 (C19); 124.16 (C18); 97.41 (Cl); 84.19 (C34). c) l-Hvdroxv-12-r2-(cvclopentvl-3-carboxaldehyde)-1methvlvinvll-23.25-dimethoxv-17-ethvl-13.19.21,27tetramethvl-ll, 28-dioxa-4-azatricvclor22.3.1.04·9loctacos18-ene-2,3,10,16-tetraone The title compound was prepared from the product of step (b) using the method of Example 1.
Example 30 l-Hvdroxv-12-F2-(cvclopentvl-3-methanol)-1-methvlvinvl15 23.25-dimethoxv-17-ethvl-13,19.21.27-tetramethvl-ll,28dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene-2,3.10,16tetraone Reduction of the product of Example 29 using the method of Example 10(a) yielded the title compound.
IQ Example 31 l-Hvdroxv-12-Γ2-(cvclopentvl-3-carboxvlic acid)-1-methvl vinyl1-23,25-dimethoxv-17-ethvl-13,19,21,27-tetramethvlll ,28-dioxa-4-azatricyclo f 223.1.04·91octacos-18-ene2,3,10,16-tetraone Oxidation of the product from Example 29 following the method of Example 12 gave the title compound.
Example 32 l-Hvdroxv-12-Γ2-fcvclopentvl-3-carboxvlic acid methyl ester)-1-methvlvinvl1-23,25-dimethoxv-17-ethvl-13,19,21,272Q tetramethvl-11.28-dioxa-4-azatricvclor22.3.1.04·9)octacos18-ene-2,3,10,16-tetraone Esterification of the product of Example 31 using the method of Example 14 yielded the title compound.
Example 33 l-Hvdroxv-12-Γ2-icvclopentvl-3-methvl propenoate)-1methvlvinvll-23,25-dimethoxv-17-ethvl-13,19,21,27tetramethvl-ll ,28-dioxa-4-azatricvclo Γ 22.3.1.04·91octacos18-ene-2,3,10.16-tetraone Wittig reaction of the product of Example 29 following the method of Example 17 gave the title compound.
Example 34 17-Allyl-l-hvdroxv-12-r2-(cvclopentvl-3-carboxaldehvde) -1-methvlvinvl1-23.25-dimethoxv-13.19.21,27-tetramethvlll .28-dioxa-4-azatricvclo Γ22.3.1.04/91octacos-18-ene2.3.10«16-tetraone The title compound was prepared from 17-allyl-l-hydroxy12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,2510 dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.04,9]octacos-l8-ene-2,3,10,16-tetraone (Example 17, WO 89/05304) using the method of Example 8(c).
Example 35 17-Allvl-l-hvdroxv-12-r2-fcvclopentvl-3-methanol)-115 methvlvinvll-23,25-dimethoxv-13.19.21.27-tetramethyl-ll.28dioxa-4-azatricvclor 22.3.1.04'9]octacos-18-ene-2,3.10,16tetraone Reduction of the product from Example 34 following the method of Example 10(a) gave the title compound. 2q Example 36 17-Allvl-l-hvdroxv-12-f2-(cvclopentvl-3-carboxvlic acid) -l-methvlvinvn-23.25-dimethoxv-13,19.21.27-tetramethvlll ,28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene2,3,10.16-tetraone Oxidation of the product of Example 34 following the method of Example 12 yielded the title compound.
Example 37 17-Allvl-l-hvdroxv-12-Γ 2-(cvc1opentvl-3-carboxylic acid methyl ester)-l-methvlvinvll-23.25-dimethoxy-13,19.21,27tetramethvl-ll ,28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos18-ene-2.3.10,16-tetraone Esterification of the product of Example 36 using the 5 method of Example 14 gave the title compound.
Example 38 17-Allvl-l-hvdroxv-12-r2-(cvclopentvl-3-methvl propenoate) -1-methvlvinvl1-23.25-dimethoxv-13,19,21.27-tetramethvl-ll.2 8-dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene2Q 2.3.10.16-tetraone Wittig reaction of the product of Example 34 following the method of Example 17 yielded the title compound.
Example 39 17-Allvl-l,14-dihvdroxv-12-Γ2-(4(S)-hydroxy-325 methoxvcvclohexvl)-1-methvlvinvl1-23,25-dimethoxv13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3.10,16-tetraone (a) 17-Allvl-l-hvdroxv-12-r2-f4-tbutvldimethvlsilvloxv3-methoxvcvclohexvl)-1-methvlvinvl1-14-^butvldimethvlsilyl 20 oxv-23.25-dimethoxv-13,19.21.27-tetramethvl-ll.28-dioxa-4azatricvclor22.3.1.04'91octacos-18-ene-2,3.10.16-tetraone The subtitle compound was prepared as in Example 5(a) (1.28g). (b) 17-Allvl-l-hvdroxv-12-Γ 2-(4-hvdroxv-325 methoxvcvclohexvl)-1-methvlvinvl1-14-^butvldimethvl silvloxv-23.25-dimethoxv-13,19,21,27-tetramethvl-ll,28dioxa-4-azatricyclo Γ 22.3.1.04'91octacos-18-ene-2,3.10.16tetraone A solution of the product from step (a) in methanol (100ml) containing pyridinium p-toluene sulphonate was stirred for 18 hours at room temperature. Volatiles were then removed in vacuo and the residue was dissolved in diethyl ether.
The ethereal solution after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine was dried (MgSO4), filtered and evaporated in vacuo to give the subtitle compound as a 2q pale yellow foam (0.97g). (c) 17-Allvl-l-hvdroxy-12-Γ 2-(4-trifluoromethylsulphonvloxv -3-methoxvcvclohexvl)-1-methvlvinvl1-14-tbutvldimethvl silyloxy-23,25-dimethoxv-13.19.21.27-tetramethvl-ll.28-dioxa -4-azatricvclor22.3.1.04'91octacos-18-ene-2.3,10,1625 tetraone To a cold (-10°C) stirred solution of the product of step (b) (0.97g) in dry dichloromethane (25ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.1ml). After stirring for 15 minutes at -10’C saturated aqueous 2q sodium hydrogen carbonate solution was added and the reaction mixture was extracted with diethyl ether. The ether extracts were then washed with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen 25 carbonate solution and brine before being dried (MgSO4), filtered and concentrated in vacuo to give the title compound as an oil (0.95g). (d) 17-Allvl-l-hvdroxy-12-r2-(4(S)-hvdroxv-3-methoxvcvclo hexvl)-1-methvlvinvl1-14-tbutvldimethvlsilvloxv-23,25dimethoxv-13,19,21.27-tetramethvl-ll,28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-2,3.10,16-tetraone Silica (55g, Merck Kieselgel 60) was added to a solution of the product of step (a) (0.9g) in dichloromethane (250 ml). Volatiles were then removed in vacuo at room temperature and the resulting freely flowing powder was stored at 8°C for 16 hours. The support was then washed with ethyl acetate and 10% acetone in ethyl acetate containing 10 2,6-dimethyl pyridine. The combined organic extracts after washing with saturated agueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine were dried (MgSO4), filtered and concentrated to an oil in lg vacuo. Chromatography on silica eluting with hexane in an acetone gradient then gave the title compound (0.28g) as a foam. (e) 17-Allvl-l.14-dihvdroxv-12-Γ2-(4(S)-hydroxy-3-methoxy cvclohexvl)-l-methvlvinvll-23.25-dimethoxv-13,19.21,27-tetra 2q methvl-11.28-dioxa-4-azatricyclor22.3.1.04·9]octacos-18ene-2.3,10.16-tetraone To a solution of the product of step (d) (0.28g) in acetonitrile (10 ml) was added 40% aqueous hydrofluoric acid (2ml). After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and the mixture was extracted with diethyl ether. The organic extract was then dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with acetone/hexane [1:2] then gave the title compound (0.22g) as a foam.
MS (FAB): 888.43 [M+Rb]+513C NMR (CDC13) 6: (Major rotamer) 212.4 (C16) 196.1 (C2); 168.9 (CIO); 164.6 (C3); 138.8 (C19) ; 135.4 (C41); 132.3 (C29); 128.9 (C31); 122.3 (C18); 116.4 (C42); 96.8 (Cl); 81.9 (C34); 77.4 (C12); 75 (C23); 73.5 (C25); 72.7 (C24); 56.8 (C9); 52.7 (C17); 48.4 (C 20); 43.3 (C15); 39.6 (C13); 39.1 (C5); 35.6 (C21); 34.6 (C27); 30.4 (C32); .9 (C7); 20.2 (C44); 13.7 (C30); 9.4 (C39).
Example 40 1.14-Dihvdroxv-12-r2-(4 fS)-hvdroxv-3-methoxvcvclohexyl)-1methvlvinvll-23.25-dimethoxy-17-ethvl-13.19,21.27-tetra methvl-11.28-dioxa-4-azatricvclor22.3.1.04·9]octacos-18ene-2.3.10.16-tetraone a) l-Hvdroxv-12-Γ2-f4 fS)-hvdroxv-3-methoxvcvclohexvl)-1methvlvinvll-14-tbutvldimethvlsilvloxv-23.25-dimethoxv-17ethvl-13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo 2q Γ22.3.1.04·91octacos-18-ene-2.3.10.16-tetraone Using the method of Example 39(a)-(d) the subtitle compound was prepared from 1,14-dihydroxy-12-[2-(4-hydroxy-3-methoxy cyclohexyl)-l-methylvinyl]-23,25-dimethoxy-17-ethyl13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (FR-900520). b) 1.14-Dihvdroxv-12-r2-(4(S)-hvdroxv-3-methoxycyclohexyl) -1-methvlvinyll-23,25-dimethoxv-17-ethvl-13.19.21.2756 tetramethvl-11,28-dioxa-4-azatricvclo Γ 22.3.1.04·91octacos18-ene-2,3,10.16-tetraone Using the method of Example 39(e) the title compound was prepared from the product of step (a).
MS (FAB): 876 [M+Rb]+ Example 41 1.14-dihvdroxv-12-r2-(4 fS)-hvdroxv-3-methoxvcvclohexvl)-1methvlvinvll-23,25-dimethoxv-17-propvl-13,19.21,27-tetra methvl-11,28-dioxa-4-azatricvclof22.3.1.04·9loctacos-18IQ ene-2,3,10,16-tetraone To a solution of the product of Example 39 (20mg) in methanol (10ml) was added 10% Pd-on-C (5mg) and the resulting suspension was then stirred in an atmosphere of hydrogen for 2 hours at O’C. The reaction mixture was then filtered and volatiles were removed in vacuo. Chromatography on silica then gave the title compound as a foam (16mg).
MS (FAB): 890 [M+Rb]+ Example 42 17-Allvl-l.14-dihvdroxv-12-Γ 2-(4-iodo-3-methoxvcvclohexvl) -1-methvlvinvl1-23,25-dimethoxv-13.19.21.27-tetramethvlll .28-dioxa-4-azatricvclo Γ 22.3.i.04'91octacos-18-ene2,3,10.16-tetraone a) 17-Allvl-l-hvdroxv-12-Γ 2-(4-iodo-3-methoxvcvclohexvl) -1-methvlvinvl1-14-fcbutvldimethvls i1vloxv-2 3.2 5-dimethoxv13.19,21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene-2.3,10.16-tetraone To a stirred, cold (-20’C) solution of the product of Example 39(d) (O.lg) in dry distilled dichloromethane (5ml) containing dry pyridine (0.4ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.3ml). After 20 minutes at -20°C 2ml of saturated aqueous sodium hydrogen carbon ate solution was added and the reaction mixture was extracted with diethyl ether. The organic extracts were then washed with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN) and saturated aqueous sodium hydrogen carbonate solution before being dried (MgSO4), filtered and concentrated to an oil in vacuo. This was taken up in dry benzene (10ml) containing triethylamine (0.1ml) and was heated under reflux for one hour. Tetra-nbutylammonium iodide (200mg) was then added and heating was continued for a further 30 15 minutes. The reaction mixture was then cooled and poured into ether. The separated ether layer was washed with dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate, sodium thiosulphate solution and brine, before being dried (MgSO4), filtered and 2Q evaporated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient gave the subtitle compound (30mg) as a foam. b) 17-Allvl-l.14-dihvdroxv-12-r2-(4-iodo-3-methoxvcvclo hexyl)-1-methvlvinyl1-23.25-dimethoxv-13.19.21.27-tetra methvl-11.28-dioxa-4-azatricvcloΓ22.3.1.04·91octacos-18ene-2,3,10,16-tetraone To a solution of the product of step (a) (30mg) in acetonitrile (7ml) was added 40% aqueous hydrofluoric acid (lml). After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and the mixture was extracted with diethyl ether. The combined ether extracts were then washed with saturated aqueous sodium hydrogen carbonate solution and brine before being dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with acetone/hexane [1:4] then gave the title compound (17mg) as a foam.
MS (FAB): 870.74 [M-I+Rb]+; 997.15 [M+Rb]+ C NMR (CDC13) δ: (Major rotamer) 213 (C16); 196.3 (C2) 169.1 (CIO); 164.8 (C3); 139.0 (C19); 135.7 (C41); 132.8 (C29); 129.1 (C31)} 122.4 (C18); 116.7 (C18); 97 (Cl); 78.9 (C34); 76.6 (C12); 75.2 (C23); 73.8 (C25); 73.0 15 (C24); 70.2 (C14); 56.7 (C9); 52.8 (C17); 26.3 (C21); 9.4 (C39).
Example 43 17-Allvl-l.14-dihvdroxv-12-r2-(3-methoxvcvclohexvl)-1methvlvinvn-23,25-dimethoxv-13.19.21.27-tetramethvl-ll.282Q dioxa-4-azatricyclor22.3.1.04'91octacos-18-ene-2,3,10,16tetraone a) 17-Allvl-l-hvdroxy-12-r2-(4-(imidazol-l-vl (thiocarbonvl)oxv)-3-methoxvcvclohexyl)-1-methvlvinvl1-14tbutvldimethvlsilvloxv-23,25-dimethoxy-13,19,21.2725 tetramethvl-11.28-dioxa-4-azatricvcloΓ22♦3.1.04'91octacos18-ene-2,3,10.16-tetraone A solution of the product of Example 39(b) (280mg) in dry distilled dichloroethane (40ml) containing - 59 's__ Ι,Ι'-thiocarbonyldiimidazole (2g) was heated under reflux for 36 hours under an atmosphere of nitrogen. Volatiles were then removed in vacuo and the residue was chromatographed on silica eluting with dichloromethane/acetone [9:1] to give the subtitle compound (105mg) as a foam. b) 17-Allvl-l,2-dihvdroxv-12-Γ 2-(3-methoxvcvclohexvl)-1methvlvinvl1-14-^butvidimethylsilvloxv-23,25-dimethoxv13.19,21.27-tetramethvl-ll.28-dioxa-4-azatricvclo IQ Γ22.3.1.04·91octacos-18-ene-3.10.16-trione A solution of the product of step (a) (105mg) in dry benzene (25ml) containing AIBN (2,2'-bisisobutyronitrile) (3mg) was heated to 40 °C under nitrogen. Tributyltin hydride (0.1ml) was then added dropwise by syringe. The temperature was then raised to 60°C over 5 minutes and a further 0.1 ml of tributyltin hydride was added. The temperature was then further raised to 90°C over 10 minutes and an additional 0.1ml of tributyltin hydride was added. After a further 10 minutes no starting material remained 2q and volatiles were removed in vacuo after cooling to room temperature. Chromatography on silica then gave the subtitle compound as an oil (85mg). c) 17-Allvl-l-hvdroxv-12-r2-(3-methoxvcvclohexvl)-1methvlvinvll-14-tbutvldimethvlsilvloxv-23.25-dimethoxv25 13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'9!octacos-18-ene-2.3.10,16-tetraone A solution of the product of step (b) (85mg) in glacial acetic acid (10ml) containing copper (II) acetate (lg) was ether. filtered heated at 80°C for 5 minutes. The cooled reaction mixture was then poured into saturated aqueous sodium hydrogen carbonate solution and this was extracted with diethyl The ether extracts were then dried (MqSO4), and concentrated to an oil in vacuo.
Chromatography on silica eluting with acetone/hexane [2:5] then gave the subtitle compound as a foam (40mg). d) 17-Allvl-l.14-dihvdroxv-12-Γ2-(3-methoxvcvclohexvl)-1methvlvinvll-23.25-dimethoxv-13.19.21.27-tetramethvl-ll,28jLq dioxa-4-azatricycloΓ22.3.1.04'91 octacos-18-ene-2,3.10,16tetraone To a solution of the product of step (c) (40mg) in acetonitrile (8ml) was added 40% aqueous hydrofluoric acid (lml). After stirrinq for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydroqen carbonate solution and the mixture was extracted with diethyl ether. The ether extracts were then dried (MqSO4), filtered and concentrated to an oil in vacuo. Chromatoqraphy on silica elutinq with hexane in an 2q increasinq acetone qradient then qave the title compound as a foam (20mq).
MS (plasma spray): [M+H-H2O]+; 788.77 [M+H]+; 13C NMR (CDC13) 6: 752.73 [M+H-2H2O]+; 805.79 [M+NH4]+ rotamer) 212.9 770.76 (C16) (Major 196.2 (C2); 169 (CIO); 164.7 (C3); 139.0 (C19); 135.6 (C41); 131.6 (C29); 130.5 (C31); 122.4 (C18); 116.7 (C42); (Cl); 78.9 (C34); 77 (C12); 75.2 (C23); 73.7 (C25); 72.8 (C24); 70.1 (C14); 56.4 (C9); 52.7 (C17); 48.5 (C20); 43.1 (C15); 39.7 (C13)? 39.2 (C5); 26.3 (C21)} 21.2 (Cl); 20.5 (C44)? 14.1 (C30); 9.4 (C39).
Example 44 1.14-dihvdroxv-12-r2-(3-methoxYcvclohexvl)-1-methvlvinvl15 23.25-dimethoxv-17-ethvl-13.19.21.27-tetramethvl-ll,28dioxa-4-azatricvclor 22♦3.1.04'91octacos-18-ene-2,3,10.16tetraone a) l-Hvdroxv-12-r2-(4-hvdroxv-3-methoxvcvclohexvl)-lmethvlvinvlΊ-14-^butvldimethvlsilvloxv-23,25-dimethoxv2Q 17-ethvl-13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene-2.3.10.16-tetraone The subtitle compound was prepared from 1,14-dihydroxy-12[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25dimethoxy-17-ethyl-13,19,21,27-tetramethyl-11,28-dioxa-425 azatricyclo[22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (FR-900520) following the method of Example 5(a) and 39(b). b) 1.14-dihvdroxv-12-Γ 2-(3-methoxycvclohexvl)-1methvlvinvll-23.25-dimethoxv-17-ethvl-13.19.21.27tetramethvl-ll. 28-dioxa-4-azatricvclor22.3.1.04'9loctacos20 18-ene-2.3.10.16-tetraone The title compound was prepared from the product of step (a) following the method of Example 43.
MS (plasma spray): 794 [M+NH4]+ Example 45 1.14-dihvdroxv-12-Γ2-f 3-methoxvcvclohexvl)-1-methvlvinyl123.25-dimethoxv-17-propyl-13,19.21.27-tetramethvl-ll,28dioxa-4-azatricvclo Γ22.3.1.04 z 9)octacos-18-ene-2,3,10,16tetraone To a solution of the product of Example 43 (28mg) in methanol (10ml) was added 10% Pd-on-C (5mg) and the resulting suspension was then stirred in an atmosphere of hydrogen for 2 hours at O’C. The reaction mixture was then filtered and volatiles were removed in vacuo. Chromatography on silica then gave the title compound as a foam (25mg).
MS (plasma spray): 808 [M+NH4]+ Example 46 IQ 17-Allyl-l-hydroxy-12-Γ 2-(3-methoxvcvclohexy1)-1-methvl vinyl1-23.25-dimethoxv-13,19.21.27-tetramethvl-ll.28-dioxa-4 -azatricvclor22.3.1.04'91octacos-18-ene-2.3.10.16-tetraone The title compound was prepared from 17-allyl-l-hydroxy-12[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25dimethoxy-13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo [22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (Example 17, WO 89/05304) following the method of Example 43.
Example 47 l-Hvdroxv-12-r2-(3-methoxvcvclohexvl)-1-methvlvinvll-23.252q dimethoxv-17-propvl-13.19.21,27-tetramethvl-ll.28-dioxa-4azatricvclo Γ22.3.1.04'9] octacos-18-ene-2.3.10.16-tetraone The title compound was prepared from l-hydroxy-12-[2(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25dimethoxy-17-propyl-13,19,21,27-tetramethyl-ll,28-dioxa-425 azatricyclo[22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (Example 12, WO 89/05304) following the method of Example 43.
Example 48 1-Hvdroxv-12-Γ2-(3-methoxvcvclohexyl)-1-methvlvinvl1-23,25dimethoxv-17-ethvl-13,19.21.27-tetramethvl-ll.28-dioxa-4azatricvclo Γ22.3.1.04,9Ί octacos-18-ene-2,3.10.16-tetraone a) 17-Ethvl-l-hydroxv-12-Γ2-(4-hvdroxv-3-methoxvcvclo 5 hexvl)-1-methvlvinvl1-23.25-dimethoxv-13,19.21.27-tetra methvl-11.28-dioxa-4-azatricvclo Γ 22.3.1.04·9loctacosa14.18-diene-2,3,10.16-tetraone 17-Ethyl-l,14-dihydroxy-12-[2-(4-hydroxy-3-methoxycyc1o hexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetra Ιθ methyl-11,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos-18ene-2,3,10,16-tetraone (FR-900520) (lOOmg) and p-toluenesulphonic acid (2mg) were dissolved in dry toluene (20ml) and were heated for 2 hours at 100°C under an atmosphere of nitrogen. Removal of solvent in vacuo and !5 chromatography on silica eluting with hexane/acetone [2:1] gave the sub-title compound as a foam (80mg).
MS (FAB): 774.8 [M+H]+; 796.85 [M+Na]+? 858.71 [M+Rb]+. 13C NMR 6: (major rotamer) 201.15 (C16); 196.0 (C2); 169.2 (CIO); 165.1 (C3); 147.8 (C15); 138.0 (C19); 123.82 (C18); 97.88 (Cl); 84.05 (C34). b) 17-Ethvl-l-hvdroxv-12-Γ2-(4-hvdroxv-3methoxvcvclohexvl)-1-methvlvinvl1-23,25-dimethoxv13,19.21,27-tetramethvl-ll.28-dioxa-4-azatricvclo 25 Γ22.3.1.04·91octacos-18-ene-2.3.10.16-tetraone A sample of the product from step (a) was dissolved in methanol (20ml) and 10% Pd-on-carbon (lOmg) was added. The mixture was stirred in an atmosphere of hydrogen for 1.5 hours at room temperature and pressure, and was then filtered through celite and evaporated to an oil in vacuo.
Column chromatography on silica eluting with hexane/acetone [2:1] gave the title compound as a foam (50mg).
MS (FAB): 776 [M+H]+? 798 [M+Na]+; 860 [M+Rb]+. 13C NMR δ: (major rotamer) 212.34 (C16); 196.42 (C2); 169.38 (CIO); 165.16 (C3); 138.9 (C19); 124.16 (C18); 97.41 (Cl); 84.19 (C34). c) l-Hvdroxv-12-Γ 2-(3-methoxvcvclohexvl)-1-methvlvinvl1- 17- Allvl-l.14-dihvdroxv-12-r2-(cvclohex-3-envl)-1methvlvinvll-23.25-dimethoxv-13.19,21,27-tetramethvlll ,28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos-18-ene2,3.10.16-tetraone 2q a) 17-Allvl-l-hvdroxv-12-Γ2-(4-iodo-3-methoxvcvclohexvl) -l-methvlvinvll-14-^butyldimethvlsilyloxv-23,25-dimethoxv13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3.10.16-tetraone To a stirred, cold (-20’C) solution of 17-allyl-l-hydroxy25 12 - [ 2 - (4 S)-hydroxy-3-methoxycyclohexyl) -1-methylviny1 ] -14 ^butyldimethylsilyloxy-23,25-dimethoxy-13,19,21,27tetramethyl-ll , 28-dioxa-4-azatricyclo[22.3.1.04'9]octacos18- ene-2,3,10,16-tetraone [the product of Example 39(d)] (0.54g) in dry distilled dichloromethane (25ml) containing dry pyridine (2ml) under nitrogen was added trifluoromethanesulphonic anhydride (1.2ml). After 20 minutes at -20°C 10ml of saturated agueous sodium hydrogen carbonate solution was added and the reaction mixture was extracted with diethyl ether. The organic extracts after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN) and saturated aqueous sodium hydrogen carbonate solution were IQ dried (MgSO4), filtered and concentrated to an oil in vacuo. This was taken up in dry benzene (30ml) containing dry pyridine (0.3ml) and tetra-nbutylammonium iodide (l.Og) was added. After heating for 30 minutes under reflux the reaction mixture was cooled to room temperature and poured into ether. The separated ether layer was washed with dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate, sodium thiosulphate solution and brine, before being dried (MgSO4), filtered and evaporated to an oil in vacuo.
Chromatography on silica eluting with acetone/hexane [1:4] then gave the title compound (500mg) as a diastereoisomeric mixture of iodides. [A smaller scale synthesis of the subtitle compound was described in Example 42(a)]. b) 17-Allvl-l.2-dihvdroxv-12-Γ 2-(cvclohex-3-envl)-12g methvlvinvll-14-tbutvldimethvlsilvloxv-23,25-dimethoxv13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-3.10.16-trione To a solution of the product of step (a) (500mg) in glacial acetic acid (8ml) was added zinc dust. After stirring for 10 minutes at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and this was extracted with diethyl ether. The ether extracts were then washed with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN) and saturated aqueous sodium hydrogen carbonate solution before being dried (MgSO4), filtered and concentrated in vacuo to give the subtitle compound (320mg) as an oil. c) 17-Allvl-l-hvdroxv-12-r2-(cvclohex-3-envl)-lmethvlvinvl1-14-tbutvldimethvlsilvloxv-23,25-dimethoxv13,19,21,27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04· 91octacos-18-ene-2.3.10,16-tetraone A solution of the product of step (b) (320mg) in glacial acetic acid (8ml) containing copper (II) acetate was heated at 85°C for 5 minutes. After cooling to room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and this was then extracted with diethyl ether. The organic extract after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN) and saturated aqueous sodium hydrogen carbonate solution was dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound as a foam (280mg). d) 17-Allvl-l.14-dihvdroxv-12-r2-ίcvclohex-3-envl)-167 methvlvinyn-23.25-dimethoxv-13.19.21,27-tetramethvl11.28-dioxa-4-azatricvclor22.3.1.04·91octacos-18-ene2.3.10,16-tetraone To a solution of the product from step (c) (280mg) in acetonitrile (20ml) was added 40% aqueous hydrofluoric acid (4ml). After stirrinq for 30 minutes at room temperature the reaction mixture was poured into saturated aqueous sodium hydroqen carbonate solution and the mixture was extracted with diethyl ether. The combined ether extracts IQ after washing with saturated aqueous sodium hydrogen carbonate solution were then dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound as a foam (0.227g).
MS (plasma spray): 720.52 [M+H-2H2O]+; 738.50 [M+H-H2O]+; 756.58 [M+H]+; 773.53 [M+NH4]+ MS (FAB): 840.81 [M+Rb]+13C NMR 5: (Major rotamer) 212.5 (C16); 196.2 (C2); 168.9 (CIO); 164.6 (C3); 138.8 (C19); 135.5 (C40); 131.4 (C31); 131.2 (C29); 126.9 (C34); 125.9 (C35); 122.4 (C18); 116.5 (C41); 96.9 (Cl); 77.4 (C12); 76.5 (C23); 73.5 (C25); 72.7 (C24); 69.9 (C14); 56.5 (C9); 52.7 (C17); 48.5 (C20); 43.4 (C15); 26.1 (C21); 20.3 (C43); 13.8 (C30); 9.4 (C38).
Example 50 1.14-Dihvdroxv-12-Γ 2-(cvclohex-3-envl)-1-methvlvinvl1-23,25dimethoxv-17-ethvl-13.19.21.27-tetramethvl-ll.28-dioxa-4azatricvclo Γ22.3.1.04'9 Ί octacos-18-ene-2.3.10,16-tetraone The title compound was prepared from the subtitle compound of Example 40(a) using the method of Example 49.
MS (FAB): 829 [M+Rb]+.
Example 51 1.14-dihydroxv-12-r2-(cvclohex-3-enyl)-1-methvlvinvl1-23,255 dimethoxv-17-propyl-13,19,21.27-tetramethvl-ll.28-dioxa-4azatricvclor22.3.1.04·91octacos-18-ene-2.3.10,16-tetraone a) 1-Hydroxy-12 - ί 2 - (4 (S)-hvdroxv-3-methoxycvc1ohexvl) 1-methvlvinvl1-14-tbutvldimethvlsilvloxv-23,25-dimethoxv17-propyl-13.19,21.27-tetramethvl-ll,28-dioxa-4-azatricvclo IQ Γ22.3.1.04·91octacos-18-ene-2.3.10,16-tetraone The subtitle compound was prepared from 1,14-dihydroxy12-[2-(4-hydroxy-3-methoxycyclohexy1) -1-methylvinyl)-23,25dimethoxy-17-propyl-13,19,21,27-tetramethyl-11,28-dioxa-4azatricyclo[22.3.1.04'9]octacos-18-ene-2,3,10,16-tetraone (Example 10, WO 89/05304) following the method of Example 39(a)-(d). b) 1.14-dihvdroxv-12-r2-(cvclohex-3-envl)-l-methvlvinvll23.25- dimethoxv-17-propvl-13.19.21.27-tetramethvl-ll.28dioxa-4-azatricvclor22. 3 ♦ 1.04'91octacos-18-ene-2.3,10.1620 tetraone The title compound was prepared from the product of step (a) following the method of Example 49.
MS (FAB): 843 [M+Rb]+ Example 52 17-Allvl-l-hvdroxv-12-Γ 2-(cvclohex-3-envl)-1-methvlvinvl123.25- dimethoxv-13,19,21.27-tetramethvl-ll.28-dioxa-4azatricvclor22.3.1.04'91octacos-18-ene-2,3.10,16-tetraone (a) 17-Allvl-l-hvdroxv-12-Γ2-(4(S)-hvdroxv-369 methoxvcvclohexvl)-1-methvlvinvl1-23,25-dimethoxv13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'9!octacos-18-ene-2.3.10.16-tetraone The subtitle compound was prepared from 17-allyl-l5 hydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl] -23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-aza tricyclo[22.3.1.04·9]octacos-18-ene-2,3,10,16-tetraone (Example 17, WO 89/05304) following the method of Example 39(a)-(d). (b) 17-Allvl-l-hvdroxv-12-Γ 2-f cvclohex-3-envl)-1-methvl vinvll-23,25-dimethoxv-13,19,21,27-tetramethvl-ll.28-dioxa-4 -azatricvclo Γ 22.3.1.04>91octacos-18-ene-2,3.10.16-tetraone The title compound was prepared from the product of step (a) following the method of Example 49(a)-(c).
Example 53 l-Hvdroxv-12-Γ2-(cvclohex-3-envl)-1-methvlvinvl1-23,25dimethoxv-17-ethvl-13.19,21,27-tetramethvl-ll,28-dioxa-4azatricvclor22.3.1.04·91octacos-18-ene-2,3.10.16-tetraone a) l-Hvdroxv-12-Γ2-(4 fS)-hvdroxv-3-methoxvcvclohexvl) 20 1-methvlvinvl1-23.25-dimethoxv-17-ethyl-13.19,21.27tetramethvl-ll .28-dioxa-4-azatricvclo Γ22.3.1.04.91octacos18-ene-2.3,10,16-tetraone The subtitle compound was prepared from 1-hydroxy12-[2-(4-hydroxy-3-methoxycyclohexyl)-1-methylvinyl]-23,2525 dimethoxy-17-ethyl-13,19,21,27-tetramethyl-11,28-dioxa-4-aza tricyclo[22.3.1.04,9]octacos-18-ene-2,3,10,16-tetraone [the product of Example 48(b)] following the method of Example 39(a)-(d).
MS (FAB): 861 [M+Rb]+ b) l-Hvdroxv-12-Γ2-(cvclohex-3-envl)-l-methvlvinvll-23,25dimethoxv-17-ethvl-13.19,21.27-tetramethvl-ll.28-dioxa-4azatricvclo Γ22.3.1.04'9!octacos-18-ene-2.3,10,16-tetraone The title compound was prepared from the product of step (a) following the method of Example 49(a)-(c).
Example 54 l-Hvdroxv-12-Γ2-(cvclohex-3-envl)-1-methvlvinvl1-23,25dimethoxv-17-propvl-13,19,21,27-tetramethvl-ll.28-dioxa-41Q azatricycloΓ22.3.1.04·91octacos-18-ene-2,3,10,16-tetraone a) l-Hvdroxv-12-Γ 2 — (4-trifluoromethylsulphonyloxv-3methoxvcvclohexvl)-l-methvlvinvll-23.25-dimethoxv-17-propvl13.19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'9!octacos-18-ene-2.3.10.16-tetraone To a cold (-10°C) stirred solution of 1-hydroxy-12-(2-(4-hydroxy-3-methoxycyclohexyi)-1methylvinyl]-23,25-dimethoxy-17-propyl-13,19,21,27tetramethyl-ll ,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos18-ene-2,3,10,16-tetraone (Example 12, WO 89/05304) (0.3g) 2Q in dry dichloromethane (12ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.1ml) until no starting material remained. Saturated aqueous sodium hydrogen carbonate solution was then added and the reaction mixture was extracted with diethyl ether. The ether extracts, after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), and saturated aqueous sodium hydrogen carbonate solution, were dried (MgSO4), filtered and concentrated in vacuo to give the subtitle compound as an oil (300mg). b) l-Hvdroxv-12-r2-(4(S)-hvdroxv-3-methoxycvclohexvl)-1methvlvinvl1-23,25-dimethoxv-17-propyl-13.19.21.27tetramethvl-ll . 28-dioxa-4-azatricvclo Γ 22.3.1.04'91octacos5 18-ene-2,3,10.16-tetraone Silica (18g, Merck Kieselgel 60) was added to a solution of the product of step (a) (300mg) in dichloromethane (100ml). Volatiles were then removed in vacuo at room temperature and the resulting freely flowing 2q powder was stored at 8°C for 16 hours. The support was then washed with acetone containing triethylamine and the solvent was evaporated in vacuo to an oil. Chromatography on silica eluting with hexane in an acetone gradient then gave the title compound as a foam (79mg).
|+ . MS (FAB): 772.83 [M+H-H2O] + ; 812.85 [M+Na]r; 874.65 [M+Rb]+ C NMR (CDC13) 6: (Major rotamer) 212.2 (C16) 196.2 (C2); 169.2 (CIO); 165.1 (C3); 138.0 (C19); 131.3 (C29); 130.2 (C31) ; 124. 1 (C18); 97 20 <C35>; 56.1 (C9) ; 53.4 (C17); 49.1 (C13); 34.5 (C27) ; 30.5 (C32); 26.3 (C41) c) l-Hvdroxv-12-Γ2-(cvclohex-3-enyl)-l-methvlvinvll-23.25dimethoxv-17-propvl-13,19.21.27-tetramethvl-ll.28-dioxa-425 azatricvcloΓ22.3.1.04'91octacos-18-ene-2,3,10,16-tetraone The title compound was prepared from the product of step (a) following the method of Example 49(a)-(c).Example 55 1,14-Dihvdroxv-12-(2-cvclohexvl-l-methvlvinvl)-23,2572 dimethoxv-17-propvl-13,19,21.27-tetramethvl-ll,28-dioxa-4azatricvclo Γ 22.3.1.04'91octacos-18-ene-2,3,10.16-tetraone To a solution of the product of Example 49 (60mg) in dry methanol (12ml) was added 10% Pd-on-C (lOOmg) and the resulting suspension was stirred in an ice bath for one hour under an atmosphere of hydrogen. The reaction mixture was then filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound as a foam (44mg).
MS (plasma spray): 724.56 [M+H-2H2O]+; 742.54 [M+H-H2O]+; 760.63 [M+H]+; 777.61 [M+NH4]+ MS (FAB): 844.86 [M+Rb]+13C NMR (CDC13) δ: (Major rotamer) 213.1 (C16) 195.9 (C2); 168.7 (CIO); 164.4 (C3); 138.0 (C19); 131.9 (C31); 130.3 (C29) ; 123 (C18); 96.7 (Ci) ; 74.9 (C23) ; 73.3 (C25); 72.5 (C24); 69.8 (C14) ; 56.3 (C9) ; 52.6 (C17) ; 48.3 (C20); 43.1 (C15) ; 39.3 (C13) ; 38.8 (C5) ; 36.2 (C32) ; 34.2 (C27); 20.1 (C43); 9.2 (C38).
Example 56 1.14-Dihvdroxv-12-r2-cvclohexvl-l-methvlvinvn-23.25dimethoxv-17-ethvl-13,19.21.27-tetramethvl-ll.28-dioxa-4azatricvclor22.3.1.04'91octacos-18-ene-2,3.10,16-tetraone To a solution of the product of Example 50 (15mg) in dry methanol (4ml) was added 10% Pd-on-C (6mg) and the resulting suspension was stirred in an ice bath for one hour under an atmosphere of hydrogen. The reaction mixture was then filtered and concentrated to an oil in vacuo.
IE 91847 Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound as a foam (14mg).
MS (FAB): 831 [M+Rb]+ Example 57 l-Hvdroxv-12-r2-cvclohexvl-l-methvlvinvl1-23.25-dimethoxv17-ethvl-13,19.21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04,9!octacos-18-ene-2,3,10,16-tetraone The title compound was prepared from the product of Example 2Q 53 following the method of Example 55.
Example 58 l-Hvdroxv-12-r2-cvclohexvl-l-methvlvinvl1-23,25-dimethoxv17-propvl-13,19,21.27-tetramethvl-ll,28-dioxa-4-azatricvclo Γ22♦3.1.04'91octacos-18-ene-2,3,10,16-tetraone The title compound was prepared from the product of Example following the method of Example 55.
Example 59 17- Allvl-l,14-dihvdroxv-12-Γ 2-(3-methoxvcvclohexvl)-1methvlvinvlt-23.25-dimethoxv-13,19,21.27-tetramethvl-ll.282q dioxa-4-azatricvcloΓ 22.3.1.04·91octacos-18-ene-3,10,16trione a) 17-Allvl-l-hvdroxv-12-Γ 2-(A-^utvldimethylsilyloxv3-methoxvcvclohexyl)-1-methvlvinvl1-14tbutvldimethvlsilvloxv-23.25-dimethoxv-13.19.21.2725 tetramethvl-11.28-dioxa-4-azatricvclor22.3.1.04·91octacos18- ene-2.3.10.16-tetraone To a cold (0’C) stirred solution of FR-900506 (lg) in dry dichloromethane (25ml) containing 2,6-dimethylpyridine IE 91847 - 74 was added under (5ml) under nitrogen tbutyldimethylsilyltriflate (2ml) until all the starting material had disappeared. The reaction mixture was then quenched with water and, after stirring for 5 minutes at room temperature, was extracted with diethyl ether. The ether extracts after washing with dilute aqueous hydrochloric acid (IN)(x2), saturated aqueous sodium hydrogen carbonate solution and brine were dried (MgSO4), filtered and concentrated in vacuo to give the subtitle compound as an oil (1.28g). b) 17-Allvl-l-hvdroxv-12-Γ 2-(4-hvdroxv-3methoxvcvclohexy1)-1-methvlvinvll-14-^butvldimethyl silvloxv-23.25-dimethoxv-13.19,21,27-tetramethvl-ll,28dioxa-4-azatricvclor22.3.1.04·91octacos-18-ene-2.3,10.16tetraone A solution of the product from step (a) in methanol (100ml) containing pyridinium p-toluene sulphonate was stirred for 18 hours at room temperature. Volatiles were then removed in vacuo and the residue was dissolved in diethyl ether. The ethereal solution after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine was dried (MgSO4), filtered and evaporated in vacuo to give the subtitle compound as a pale yellow foam (0.97g). c) 17-Allyl-l-hvdroxv-12-(2-(4-(imidazol-l-yl (thiocarbonvl1oxv)-3-methoxvcvclohexvl1-1-methvlvinvll-14^butvldimethvlsilvloxv-23,25-dimethoxv-13,19,21,27IE 91847 - 75 10 tetramethvl-11.28-dioxa-4-azatricyclor22.3.1.04·91octacos18-ene-2.3,10,16-tetraone A solution of the product of step (b) (280mg) in dry distilled dichloroethane (40ml) containing 1,1'-thiocarbonyldiimidazole (2g) was heated under reflux for 36 hours under an atmosphere of nitrogen. Volatiles were then removed in vacuo and the residue was chromatographed on silica eluting with dichloromethane/acetone [9:1] to give the subtitle compound (105mg) as a foam. d) 17-Allvl-l.2-dihvdroxv-12-r2-(3-methoxvcvclohexvl)-1methvlv i nvl1-14-^butvldimethvlsi1vloxy-23.25-d imethoxy13.19.21,27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-3.10,16-trione A solution of the product of step (c) (105mg) in dry benzene (25ml) containing AIBN (2,2’-bisisobutyronitrile) (3mg) was heated to 40 °C under nitrogen. Tributyltin hydride (0.1ml) was then added dropwise by syringe. The temperature was then raised to 60°C over 5 minutes and a further 0.1 ml of tributyltin hydride was added. The temperature was then further raised to 90°C over 10 minutes and an additional 0.1ml of tributyltin hydride was added. After a further 10 minutes no starting material remained and volatiles were removed in vacuo after cooling to room temperature. Chromatography on silica then gave the subtitle compound as an oil (85mg). e) 17-Allvl-l-hvdroxv-12-r2-(3-methoxvcyclohexvl)-1methvlvinvl1-14-^butvldimethvlsilvloxv-2 3,25-dimethoxvIE 91847 - 76 13,19,21,27-tetramethvl-ll,28-dioxa-4-azatricyclo Γ 22♦3.1.04'91octacos-18-ene-2.3.10.16-tetraone A solution of the product of step (d) (85mg) in glacial acetic acid (10ml) containing copper (II) acetate (lg) was heated at 80°C for 5 minutes. The cooled reaction mixture was then poured into saturated agueous sodium hydrogen carbonate solution and this was extracted with diethyl ether. The ether extracts were then dried (MgSO4), filtered and concentrated to an oil in vacuo.
IQ Chromatography on silica eluting with acetone/hexane [2:5] then gave the subtitle compound as a foam (40mg). f) 17-Allvl-l,14-dihvdroxv-12-Γ 2-(3-methoxvcvclohexvl)-1methvlvinvll-23.25-dimethoxv-13,19.21.27-tetramethvl-ll.28dioxa-4-azatricvclor22.3.1.04'91octacos-18-ene-2.3.10.1615 tetraone To a solution of the product of step (e) (40mg) in acetonitrile (8ml) was added 40% agueous hydrofluoric acid (lml). After stirring for 1 hour at room temperature the reaction mixture was poured into saturated aqueous sodium hydrogen carbonate solution and the mixture was extracted with diethyl ether. The ether extracts were then dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the subtitle compound as a foam (20mg).
MS (plasma spray): 752.73 [M+H-2H2O] 805.79 [M+NH4]+ 770.76 (Major rotamer) 212.9 (C16); [M+H-H2O]+; 788.77 [M+H]+; 13C NMR (CDC13) 6: ΊΊ 196.2 (C2); 169 (CIO); 164.7 (C3); 139.0 (C19); 135.6 (C41); 131.6 (C29); 130.5 (C31); 122.4 (C18); 116.7 (C42) ; (Cl); 78.9 (C34); 77 (C12); 75.2 (C23); 73.7 (C25); 72.8 (C24); 70.1 (C14); 56.4 (C9); 52.7 (C17); 48.5 (C20); 43.1 (C15); 39.7 (C13); 39.2 (C5); 26.3 (C21); 21.2 (C7); 20.5 (C44); 14.1 (C30); 9.4 (C39). g) 17-Allvl-l.14-dihvdroxv-12 - Γ 2 - (3-methoxvcvclohexy1)-1methvlv invl1-2 3,2 5-dimethoxv-13,19,21.27-1etramethv1-11.28dioxa-4-azatricvclo Γ 22.3.1.04·91octacos-18-ene-3,10,16trione Hydrogen sulphide gas was bubbled through a solution of the product of step (f) (40mg) in pyridine (2ml) and dimethylformamide (0.1ml) for 2 hours at room temperature. After standing for 4 hours at room temperature dilute aqueous hydrochloric acid was added and the reaction mixture was extracted with ethyl acetate. The ethyl acetate extract was then dried (MgSO4), filtered and concentrated in vacuo. Chromatography on silica eluting with ethyl acetate then gave the title compound as a foam (25mg).
MS (FAB): 858 (M+Rb)+; 796 (M+Na)+; 774 (M+H)+; 756 (M-0H)+13C NMR (CDC13) 6: 214.3 (C16); 174 (C3); 169.4 (CIO); 141.2 (C19); 135.4 (C41); 131.6 (C29); 129.8 (C31); 121.4 (C18); 116.6 (C42); 97.8 (Cl); 78.9 (C34); 48.4 (C20); 20.7 (C7); 14.3 (C30); 9.7 (C39) Example 60 17-Al1vl-14-hvdroxv-12-Γ 2-(4-hvdroxv-3-methoxycyc1ohexvl) -1IE 91847 methvlvinvll-23,25-dimethoxv-13.19.21,27-tetramethvl-ll,28dioxa-4-azatricvclor22.3.1.04/9loctacos-18-ene-2.3.10.16tetraone and 5 17-Al1vl-14-hvdroxv-12-Γ 2-f 4-hvdroxv-3-methoxycyclohexyi)-1methvlvinvl1-23.25-dimethoxv-l.13.19.21.27-pentamethyl11,28-dioxa-4-azatricvclo Γ22.3.1.04'9)octacos-18-ene2,3,10,16-tetraone a) 17-Allvl-l-chloro-14-hvdroxv-12-r2-(4-hvdroxv-31Q methoxycyclohexyi)-l-methvlvinvll-23,25-dimethoxv13,19.21,27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'9)octacos-18-ene-2,3.10.16-tetraone A solution of FR-900506 (500mg) in dry dichloromethane (25ml) was added dropwise over 1 minute to a stirred, cool (O’C) solution of thionyl chloride (0.45ml) and pyridine (1.11ml) in dry dichloromethane (20ml) under nitrogen. After 20 minutes, saturated aqueous sodium hydrogen carbonate solution was added and the mixture was stirred at room temperature for 20 minutes. The organic extract was 2q then separated and washed with dilute aqueous hydrochloric acid (1M, 20ml), water (20ml) and brine (10ml) before being dried (MgSO4), filtered and evaporated in vacuo to give the sub-title compound as an an oil (512mg). b) 17-Al1vl-14-hvdroxv-12 - Γ 2 - f 4-hydroxy-3-methoxvcvclo hexvl)-1-methvlvinvll-23,25-dimethoxv-l3,19.21.27tetramethvl-ll .28-dioxa-4-azatricvclo Γ 22.3.1.04'9]octacos18-ene-2,3.10,16-tetraone and IE 91847 - 79 17-Allyl-14-hvdroxv-12-Γ2-(4-hvdroxv-3-methoxvcvclohexvl)-1methvlvinvll-23.25-dimethoxv-l.13.19.21.27-pentamethvl11,28-dioxa-4-azatricvclor 22.3.1.04'91octacos-18-ene2,3,10.16-tetraone To a cold (-50°C), stirred suspension of copper (I) iodide (463mg) in dry diethyl ether (20ml) under nitrogen was added a dilute (1.1M) solution of methyl lithium in ether (4.42ml). After stirring for 30 minutes at -40°C the reaction mixture was cooled to -70°C and a solution of the 2Q product from step (a) (400mg) in dry ether (20ml) was added dropwise. After stirring for 20 minutes, saturated aqueous ammonium chloride solution was added and the reaction mixture was allowed to warm to room temperature. The ethereal layer was then separated and was washed with water (20ml) and brine (20ml) before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica then gave a first isomer of the first title compound (Isomer A, 5mg), a second isomer of the first title compound (Isomer B, 20mg), and the second title compound (4.5mg).
MS (FAB): Isomer A - 770.8 [M+H-H2O]+; 788.8 [M+H]+; 810.8 [M+Na]+; 872.6 [M+Rb]+ Isomer B - 872.4 [M+Rb]+ 2nd title compound - 784.8 [M+H-H20]+; 802*8 [M+H]+; 824.8 [M+Na]+; 886.5 [M+Rb]+ —C NMR (CHC13) 6: Isomer A 211.4 (C16); 200.7 (C2); 169 (CIO); 165.6 (C3); 139.6 (C19); 135.7 (C41); 131.9 (C31); 131.2 (C29); 122.4 (C18) ; 116.5 (C42); 84.2 (C34); 80.5 (C12); 78.3 (Cl) ; 76.9 (C23); 75.2 (C24); 74.9 (C25); 73.5 (C35); 68.5 (C14) ; 53.4 (C17); 52 (C9); 47.7 (C20); 45.5 (C15); 44.3 (C5) ; 40.1 (C13); 35.2 (C40); 34.9 (C32); 34.8 (C22); 34.6 (C33) ; 32.7 (C26); 31.5 (C27); 31.2 (C36); 30.5 (C37); 27.1 (C21) ; 25.8 (C8); 24.9 (C6); 20. 8 (C7); 20.5 (C44); 17.1 (C43) ; 16.4 (C47); 13.3 (C30); 10.1 (C39) Isomer Β - 213.2 (C16) ; 197 (C2); 170.2 (CIO); 163.8 1Q (C3); 137.3 (C19); 135.2 (C41); 131.9 (C29); 128.5 (C31); 123.4 (C18); 116.7 (C42); 84.1 (C34); 83.5 (Cl); 79.3 (C12); 70.2 (C14); 55.9 (C9); 51.9 (C17); 49.4 (C20); 44.7 (C15); 40 (C5); 40.1 (C13) ; 38.5 (C40); 10.1 (C39) 2nd title compound - 212.4 (C16); 203.3 (C2); 169.4 15 (CIO); 167 (C3); 139.1 (C19); 135.6 (C41); 131.8 (C29); 129.7 (C31); 123 (C18); 116.6 (C42); 84.2 (C34); 82.9 (Cl); 77.3 (C12); 69.7 (C14); 52.5 (C17); 52 (C9); 47.7 (C20); 45.2 (C15); 44 (C5); 39.9 (C13); 14.1 (C48); 10 (C39). 2Q Isomers A and B differ in their stereochemistry at Cl. Example 61 17-Allvl-l.14-dihvdroxv-12-r2-(cvclopentvl-3-methanol(methyl ether))-1-methvlvinvl1-23,25-dimethoxv-13.19,21,27-tetra methvl-11.28-dioxa-4-azatricvclor 22.3.1.04·9loctacos-1825 ene-2.3,10,16-tetraone To a solution of the compound of Example 10(a) (73mg) in diethyl ether (2ml) containing boron trifluoride diethyl etherate (0.1ml) was added an ethereal solution of diazomethane. After standing for 30 minutes at room temperature volatiles were removed in vacuo and the residue was chromatographed on silica eluting with hexane/acetone [4:1] to give 17-allyl-l-hydroxy-12-[2-(cyclopentyl5 3-methanol(methylether))-1-methylvinyl]-23,25-dimethoxy-14tbutyldimethylsilyloxy-lS,19,21,27-tetramethyl-ll,28-dioxa -4-azatricyclo[22.3.1.04'9]octacos-18-ene-2,3,10,16tetraone (20mg) as a foam. This was dissolved in acetonitrile (5ml) and 40% agueous hydrofluoric acid Ιθ (0.5ml) was then added. After stirring for 75 minutes at room temperature the reaction mixture was poured into ethyl acetate and was washed with saturated aqueous sodium hydrogen carbonate solution and brine before being dried, (MgSO4), filtered and evaporated to an oil in vacuo.
Chromatography on silica eluting with acetone/hexane [1:3] then gave the title compound (lOmg) as a foam. 13C NMR (CDC13) 6: (Major rotamer) 213.8 (C16); 196.2 (C2); 168.9 (CIO); 164.9 (C3); 138.9 (C19); 135.6 (C40); 122.5 (C18); 116.6 (C41); 97 (Cl); 77.4 (C12); 75.2 (C23); 2Q 70.1 (C14); 58.8 (cyclopentylCH2OCH3); 56.3 (C9); 52.8 (C17); 48.6 (C20); 29.7 (C8); 26.3 (C21); 24.6 (C6); 21.1 (C7); 20.4 (C43); 14.1 (C30); 9.5 (C38) MS (FAB): 872 [M+Rb]+; 810 [M+Na]+; 788 [M+H]+.
Example 62 17-Allvl-l.14-dihvdroxv-12-r2-(4-amino-3-methoxvcvclohexvl)l-methvlvinvll-23.25-dimethoxv-13,19.21,27-tetramethvlll ,28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene2,3,10,16-tetraone a) 17-Al1ν1-1-hvdroxv-12-Γ 2-(4-a z ido-3-methoxvcyc1ohexyl) 1-methvlvinvl1-23.25-dimethoxv-14-tbutvldimethvlsilvloxv13.19.21.27- tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3.10.16-tetraone To a stirred, cold (-20°C) solution of the product of Example 39(b) (0.19g) in dry distilled dichloromethane (7ml) containing dry pyridine (0.63ml) under nitrogen was added trifluoromethanesulphonic anhydride (0.41ml). After 20 minutes at -20 °C saturated aqueous sodium hydrogen !θ carbonate solution (3ml) was added and the reaction mixture was extracted with diethyl ether. The organic extracts were then washed with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), and saturated aqueous sodium hydrogen carbonate solution ^5 before being dried (MgSO4) , filtered and concentrated to an oil in vacuo. This material was dissolved in dry DMF (5ml) and sodium azide (0.5g) was added. After stirring for 30 minutes at room temperature the reaction mixture was poured into water and this was then extracted with ethyl acetate. The organic extract after washing with brine was dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica then gave the subtitle compound (83mg) as a foam. b) 17-Allvl-l-hvdroxy-12-r2-f4-amino-3-methoxvcvclohexvl)25 1-methvlvinvl1-23.25-dimethoxv-14-tbutvldimethvlsilvloxv13.19.21.27- tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04'91octacos-18-ene-2.3.10.16-tetraone To a stirred solution of the product of step (b) (50mg) in dry, distilled methanol (5ml) under nitrogen was added 1,3-propanedithiol (0.03ml) and triethylamine (0.04ml). After stirring for 1 hour at room temperature the reaction mixture was columned on silica eluting with hexane/acetone 5 [3:1] to give the subtitle compound as a foam (37mg). 13C NMR (CDC13) 5: (Major rotamer) 209.6 (C16)? 196.5 (C2); 169.1 (CIO); 164.7 (C3); 138.5 (C19); 135.7 (C41); 133.3 (C29); 128.3 (C31); 123.2 (C18); 116.6 (C42); 97.6 (Cl); 82.4 (C34); 56.4 (C9); 53.7 (C17); 49.3 (C20); 43.7 10 (C15); 40 6 (C13); 39.2 (C5); 10.5 (C39).
MS (FAB): 1001.6 [M+Rb]+ c) 17-Allvl-l,l4-dihvdroxv-12-r 2-(4-amino-3-methoxvcvclo hexvl)-1-methvlvinvl1-23.25-dimethoxv-13,19.21.27-tetra methvl-11.28-dioxa-4-azatricvclor 22. 3.1.04'91octacos-1815 ene-2,3,10,16-tetraone To a solution of the product of step (b) (35 mg) in acetonitrile (7ml) was added 40% aqueous hydrofluoric acid (0.5ml). After stirring for 2.5 hours at room temperature the reaction mixture was poured into ethyl acetate and the 2Q separated organic extract was then washed with saturated aqueous sodium hydrogen carbonate solution and brine before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Column chromatography on silica eluting with hexane/acetone [2:1] then gave the title compound (15mg) as 2g a foam. 13C NMR (CDC13) 6: (Major rotamer) 212.9 (C16); 196.2 (C2); 169.1 (CIO); 164.8 (C3); 139.1 (C19); 135.7 (C41); 132.9 (C29); 128.5 (C31); 122.6 (C18); 116.8 (C42); 97.2 (Cl); 82.9 (C34); 78 (C12); 75.4 (C23); 73.8 (C25); 73.0 (C2 4); 70.2 (C14); 57.1 (C9); 53.1 (C17); 48.7 (C20); 43.3 (C15); 39.8 (C13); 39.4 (C5)} 24.1 (C6); 21.3 (C7); 20.6 (C44); 14.2 (C30); 9.7 (C39).
MS (FAB): 888.5 [M+Rb]+; 826.7 [M+Na]+; 786.7 [M+H-H2O]+ Example 63 17-Allvl-l.14-dihvdroxv-12-r2-(4-acetamido-3-methoxvcvclo hexvl)-l-methylvinvll-23.25-dimethoxv-13.19.21,27-tetra 10 methvl-11.28-dioxa-4-azatricvcloΓ22.3.1♦04·9loctacos-18ene-2,3,10,16-tetraone a) 17-Allvl-l-hvdroxv-12-Γ 2-(4-acetamido-3-methoxvcvclo hexvl)-l-methylvinvll-23.25-dimethoxv-14-^butyldimethvl silvloxv-13,19,21.27-tetramethvl-ll.28-dioxa-4-azatricvclo Γ22.3.1.04·91octacos-18-ene-2.3,10.16-tetraone To a solution of the product of Example 62(b) (20mg) in dry dichloromethane (3ml) was added pyridine (0.1ml) and acetyl chloride (0.1ml). After stirring for 10 minutes at room temperature the reaction mixture was poured into water and 2Q this was then extracted with diethyl ether. The organic extract was then washed with dilute aqueous hydrochloric acid and brine before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane/acetone [3:1] then gave the subtitle compound (15mg) as an oil. b) 17-Allvl-l.14-dihvdroxv-12-r2-(4-acetamido-3-methoxy cvclohexvl)-l-methvlvinvn-23,25-dimethoxv-13.19.21.27-tetra methvl-11.28-dioxa-4-azatricvclor22.3.1.04·9loctacos-1885 ene-2,3.10.16-tetraone A portion of the product from step (a) (13mg) was dissolved in acetonitrile (4ml) and to this was added 40% aqueous hydrofluoric acid (0.1ml). After stirring for 2 hours at 5 room temperature the reaction mixture was poured into ethyl acetate and the separated organic extract was then washed with water, saturated aqueous sodium hydrogen carbonate solution and brine before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Column chromatography 2q on silica eluting with hexane/acetone [2:1] then gave the title compound (8mg) as a foam. 13C NMR (CDC13) δ: (Major rotamer) 212.4 (C16); 196.2 (C2) ; 169 (CIO); 164.7 (C3) ; 139 (C19) ; 135.5 (C41) ; 122.4 (C18); 116.7 (C42); 97 (Cl); 9.4 (C39) 153H NMR (CDC13) 6: 2.01 [3H,s,NHCOCH3] MS (FAB): 930.5 [M+Rb]+; 868.9 [M+Na]+ Example 64 17-Allvl-l.14-dihvdroxv-12-r2-(4-formvloxv-3-methoxvcvclo hexvl)-l-methvlvinvll-23,25-dimethoxv-l3.19,21,27-tetra 2q methvl-11.28-dioxa-4-azatricvcloΓ22.3.1.04'91octacos-18ene-2.3.10.16-tetraone To a solution of the compound of Example 39(c) (1.103g) in dry DMF (20ml) was added sodium azide (2.58g). After stirring for 2 hours at room temperature the reaction mixture was poured into water and this was then extracted with ethyl acetate. The organic extract after washing with brine was dried (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane/acetone [3:1] then gave 17-allyl-l-hydroxy-12-[2-(4formyloxy-3-methoxycyclohexyl)-1-methylvinyl]-23,25dimethoxy-14-tbutyldimethylsilyloxy-13,19,21,27-tetra methyl-11,28-dioxa-4-azatricyclo[22.3.1.04'9]octacos-185 ene-2,3,10,16-tetraone (115mg) as a foam. A portion of this (71mg) was dissolved in acetonitrile (14ml) and to this was added 40% aqueous hydrofluoric acid (0.5ml). After stirring for 3.5 hours at room temperature the reaction mixture was poured into ethyl acetate and the IQ separated organic extract was then washed with water, saturated aqueous sodium hydrogen carbonate solution and brine before being dried (MgSO4), filtered and evaporated in vacuo to an oil. Column chromatography on silica eluting with hexane/acetone [2:1] then gave the title compound (19mg) as a foam. 13C NMR (CDC13) 6: (Major rotamer) 212.7 (C16); 196.2 (C2); 169.2 (CIO); 164.8 (C3); 160.6 (OCHO-); 138.9 (C19); 135.5 (C41); 132.4 (C29); 129.5 (C31); 122.4 (C18); 116.6 (C42); 96.9 (Cl); 78.7 (C34); 77.3 (C12); 75.1 (C23); 72.8 (C24); 70 (C14); 56.6 (C9); 52.7 (C17); 48.5 (C20); 43 (C15); 39.6 (C13); 39.2 (C5); 28.2 (C8); 26.2 (C21); 24.5 (C6); 21.1 (C7); 20.4 (C44); 14.1 (C30); 9.3 (C39).
MS (FAB): 916.2 [M+Rb]+; 854.5 [M+Na]+; 832.6 [M+H]+; 814.6 [M+H-H2O]+ Example 65 17-Allvl-l.14-dihvdroxv-12-Γ2-f3-oxo-cvclohexvl)-1-methvl vinvll-23,25-dimethoxv-13,19,21.27-tetramethvl-ll,28-dioxa4-azatricvcloΓ22.3.1.047 91octacos-18-ene-2,3,10,1687 tetraone and 17-Allvl-l.14-dihvdroxv-12-Γ2-(3-methoxv-cvclohex-4-envl)-1methvlvinvll-23.25-dimethoxv-l3.19,21.27-tetramethvl-ll.285 dioxa-4-azatricvcloΓ22.3.1.04·91octacos-18-ene-2.3,10,16tetraone Silica (220g, Merck Kieselgel 60, Art. 15111) was added to a solution of the compound of Example 39(c) (250ml). Volatiles were then removed in vacuo at room temperature IQ and the resulting freely flowing powder was stored at 8°C for 16 hours. The support was then washed with ethyl acetate and 10% acetone in ethyl acetate containing 2,6-dimethylpyridine. The combined organic extracts after washing with saturated aqueous sodium hydrogen carbonate solution, dilute aqueous hydrochloric acid (IN), saturated aqueous sodium hydrogen carbonate solution and brine were dried, (MgSO4), filtered and concentrated to an oil in vacuo. Chromatography on silica eluting with hexane in an acetone gradient then gave the compound of Example 39(d) 2Q (1.12g) as a foam. Further elution then gave the compound of Example 8(c) (0.5g) as a foam.
Mixed fractions were then combined, treated with 40% aqueous hydrofluoric acid as above, and re-chromatographed on silica eluting with ethyl acetate to give the first title compound (200mg). 13C NMR (CDC13) 5: (Major rotamer) 212.5 (C16); 210.7 (C34)? 196.2 (C2); 169 (CIO); 164.7 (C3); 139 (C19); 135.5 (C41); 133.1 (C29) ; 129 (C31); 122.5 (C18); 116.7 (C42); 97.1 (Cl); 77.6 (C12); 75.2 (C23); 73.7 (C25); 72.8 (C 24); 69.9 (C14); 56.3 (C9); 52.9 (C17); 48.6 (C20); 47.6 (C33) ; 43.5 (C15) ; 41.2 (C35); 39.7 (C13); 37.9 (C32) ; 26.2 (C21); 25.8 (C8) ; 24.5 (C6); 21.1 (C7); 20.4 (C44) ; 13.8 (C30) ; 9.7 (C39).
MS (FAB): 856 [M+Rb]+; 794 [M+Na]+; 736 [M+H-2H2O]+ Further elution then gave the second title compound. 13 NMR (CDC13): S (C2); 168.9 (CIO); 132.3 (C29); 129.9 (C35); 128.1 (C36); (C14); 56.5 (C9); (C21); 24.4 (C6); 21 (Major rotamer) 212.6 (C16); 196.2 164.6 (C3); 139.7 (C19); 135.5 (C41); (C31); 122.3 (C18); 116.5 (C42); 128.5 96.9 (Cl); 73.5 (C25); 72.7 (C24); 70.5 52.7 (C17); 48.4 (C20); 27.6 (C8); 26.1 (C7); 20.3 (C44); 14 (C30); 9.3 (C39).
MS (FAB): 870 [M+Rb]+; 808 [M+Na]+ Example 66 17-Allvl-l.14-dihvdroxv-12-r2-(cvclopentvl-3-carboxylic acid morpholine amide)-l-methvlvinvll-23.25-dimethoxv -13,19.21.27-tetramethvl-ll,28-dioxa-4-azatricvclo 2Q Γ22.3.1.04'9)octacos-18-ene-2.3.10,16-tetraone To a solution of the product of Example 13 was added morpholine (0.03ml) followed by triethylamine (0.03ml) and 2-chloro-l-methylpyridinium tosylate (70mg). After stirring for 1 hour at room temperature a further portion of the tosylate (40mg) was added and stirring was continued for 5.5 hours at room temperature. Additional triethylamine (0.03ml) and morpholine (0.03ml) was then added and the reaction mixture was stirred overnight at room temperature. The reaction was then quenched with dilute aqueous hydrochloric acid (2M, 10ml) and the mixture was extracted with ethyl acetate. The organic extracts were then washed with saturated aqueous sodium hydrogen 5 carbonate solution and brine before being dried (MgSO4), filtered and evaporated to an oil in vacuo. Chromatography on silica eluting with hexane in an increasing acetone gradient then gave the title compound (30mg) as a foam.
MS (FAB): 941.4 [M+Rb]+; 880.2 [M+Na]+; 858.4 [M+H]+; 1Q 840.4 [M+H-H2O]+13C NMR (CDC13) 5: (Major rotamer) 212.4 (C16); 196.2 (C2); 174.6 (cyclopentylCO); 169.1 (CIO); 164.7 (C3); 138.9 (C19); 135.6 (C40); 132.5 (C29); 131.3 (C31); 122.7 (C18); 116.7 (C41); 97.1 (Cl); 70.0 (C14); 67 and 66.8 (morpholine 15 CH2O); 56.3 (C9); 52.9 (C17); 48.8 (C20); 46.1 and 42.3 (morpholineCH2N); 27.8 (C8); 26.2 (C21); 24.5 (C6); 21.0 (C7); 20.3 (C43); 14.1 (C30); 9.9 (C38)

Claims (18)

CLAIMS:
1. A compound of formula I, wherein R 1 represents H, OH or alkoxy; R 2 represents H; in addition, R 1 and R 2 may together represent a second bond between the carbon atoms to which they are attached; R 3 represents methyl, ethyl, propyl or allyl; R 4 represents H, OH, alkyl, alkoxy, halogen, amino, S-alkyl, NHCHO or NHCO-alkyl; n represents 1 or 2; X represents 0, (H,OH), (H,H) or =NH; and Y represents a cyclic group of formula II, in which R 5 represents (Η,Η), (Η,ΟΗ), (H,methoxy) or O; R 6 represents H, (R)-OH, (S)-OH, alkoxy, amino, alkylamino, alkanoylamino, formyloxy or halogen; R 7 represents H; and in addition R 5 and R 6 may together 5 represent a second bond between the carbon atoms to which they are attached; or R 6 and R 7 may together represent a second bond between the carbon atoms to which they are attached; or a cyclic group of formula III, III 15 in which R 8 represents alkyl substituted by one or more groups selected from OH, alkoxy, =0, and CO 2 H; or alkenyl optionally substituted by one or more groups selected from OH, =0, or CO 2 H; provided that
2. Q a) when n represents 1; R 1 represents OH; R 3 represents allyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 represents (R)-OH; then X does not represent O; b) when n represents 2; R x represents OH; R 4 i) R J represents methyl, ethyl, allyl or propyl; R 4 represents OH; R 5 represents (H,methoxy) and R 6 represents (R)-OH; then X does not represent O; ii) when R 1 and R 2 together represent a second bond between the carbon atoms to which they are attached or each represent H; R 3 represents allyl or propyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 5 represents (R)-OH; then X does not represent 0; iii) when R 1 represents OH, methoxy or together with R 2 it represents a second bond between the carbon atoms to which they are attached; R 3 represents allyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 IQ represents methoxy; then X does not represent 0; iv) when R 1 represents H or OH; R 3 represents allyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 represents (R)-OH; then X does not represent (H,OH); v) when R 1 represents H; R 3 represents propyl; R 4 15 represents OH; R 5 represents (H,OH); and R 6 represents (R)-OH; then X does not represent 0; vi) when R 1 represents OH; R 3 represents ethyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 represents (R)-OH; then X does not represent (H,OH); 2Q vii) when R 1 and R 2 together represent a second bond between the Carbon atoms to which they are attached or each represent H; R 3 represents ethyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 represents (R)-OH; then X does not represent 0; 25 viii) when R 1 represents OH; R 3 represents allyl; R 4 represents OH; R 5 represents (H,OH) or (H,methoxy); and R 6 represents (R)-OH; then X does not represent (H,H); ix) when R 1 represents OH; R 3 represents ethyl; R 4 represents OH; R 5 represents (H,methoxy); and R 6 represents (R)-OH; then X does not represent (H,H); x) when R 1 represents OH; R 3 represents methyl, ethyl or allyl; R 4 represents OH; R 5 represents (H,OH); and 5 R 6 represents (R)-OH; then X does not represent 0; and xi) when R 1 represents OH; R 3 represents allyl; R 4 represents OH; R 5 represents 0; and R 6 represents (R)-OH; then X does not represent 0; and pharmaceutically acceptable derivatives thereof. Ιθ 2. A compound of formula I, as claimed in claim 1, wherein R 1 represents H or OH.
3. A compound of formula I, as claimed in claim 1 or claim 2, wherein R 4 represents H, OH, alkyl, halogen or amino. 15
4. A compound of formula I, as claimed in any one of the preceding claims, wherein R 5 represents (H,0H) or (H,methoxy).
5. A compound of formula I, as claimed in any one of the preceding claims, wherein R 6 represents H, (R)-OH or 2θ amino.
6. A compound of formula I, as claimed in any one of the preceding claims, wherein R 8 represents an amide of a CO 2 H group or alkyl substituted by alkoxy.
7. A compound of formula I, as claimed in claim 1, which 25 is: 17-allyl-l,14-dihydroxy-12-(2-(3-methoxycyclohexyl)-1methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-ll,28dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,1694 tetraone; 17-allyl-l,14-dihydroxy-12-[2-(cyclopentyl-3-carboxylic acid morpholine amide)-1-methylvinyl]-23,25-dimethoxy13,19,21,27-tetramethyl-ll,28-dioxa-4-azatricyclo 5 [22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16-tetraone; 17-allyl-14-hydroxy-12-(2-(4-hydroxy-3-methoxyeye1ohexy1)-1methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-ll,28dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene-2,3,10,16tetraone ; IQ 17-allyl-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyclohexyl)-1methylvinyl]-23,25-dimethoxy-l,13,19,21,27-pentamethyl11,28-dioxa-4-azatricyclo[22.3.1.0 4,9 ]octacos-18-ene2,3,10,16-tetraone; 17-allyl-l-amino-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyclo 15 hexyl)-1-methylvinyl]-23,25-dimethoxy-l3,19,21,27-tetra methyl-11,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18ene-2,3,10,16-tetraone; 17- allyl-l-fluoro-14-hydroxy-12-[2-(4-hydroxy-3-methoxycyclo hexyl) -1-methy1vinyl]-2 3,2 5-dimethoxy-13,19,21,2720 tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos18- ene-2,3,10,16-tetraone; 17-Allyl-l,14-dihydroxy-12-[2-(cyclopentyl-3-methanol(methyl ether))-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetra methyl-11,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-1825 ene-2,3,10,16-tetraone; or 17-Ally1-1,14-dihydroxy-12-[2-(4-amino-3-methoxycyclohexy1) 1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethylll ,28-dioxa-4-azatricyclo[22.3.1.0 4 ' 9 ]octacos-18-ene95 2,3,10,16-tetraone.
8. The use of a compound of formula I, as defined in claim 1, as a pharmaceutical.
9. A pharmaceutical composition comprising a compound of 5 formula I, as defined in claim 1, in association with a pharmaceutically acceptable adjuvant, diluent or carrier.
10. The use of a compound of formula I, as defined in claim 1, in the manufacture of a medicament for use as an immunosuppressive agent. IQ
11. A method of effecting immunosuppression which comprises administering a therapeutically effective amount of a compound of formula I, as defined in claim 1, to a patient.
12. A process for the production of a compound of formula 15 I as defined in claim 1, which comprises: (a) producing a compound of formula I in which R 1 and r 2 together represent a second carbon-carbon bond between the carbon atoms to which they are attached, by dehydration of a corresponding compound in which R 1 represents OH and 2q R 2 represents H; (b) producing a compound of formula I in which R 3 · and R 2 each represent hydrogen, by reduction of a corresponding compound in which R and R together represent a second carbon-carbon bond between the carbon 25 atoms to which they are attached; (c) producing a compound of formula I in which X represents (Η,ΟΗ), by reduction of a corresponding compound in which X represents 0; (d) producing a compound of formula I in which X represents (H,H), by reduction of a corresponding compound in which X represents 0; (e) producing a compound of formula I in which X 5 represents 0, by oxidation of a corresponding compound in which X represents (H,0H); (f) producing a compound of formula I in which R 4 represents alkoxy, by reaction of a corresponding compound in which R 4 represents OH and X represents (H,OH) with an IQ alkanol; (g) producing a compound of formula I in which R 4 represents halogen, by reaction of a corresponding compound in which R 4 represents OH with a suitable halogenating agent; (h) producing a compound of formula I in which R 4 represents H or alkyl, by reaction of a corresponding compound in which R 4 organometallic reagent; represents halogen with an (i) producing compound of formula I in which 2Q represents amino, by reaction of a corresponding compound in which R 4 represents halogen with ammonia; (j) producing a compound of formula I in which X represents =NH, by reaction of a corresponding compound in which R 4 represents halogen with ammonia; 25 (k) producing a compound of formula I in which R 4 represents S-alkyl, by reaction of a corresponding compound in which R 4 represents halogen with an alkylthiol; (1) producing compound of formula I in which R - 97 *r represents NHCHO, by reaction of a corresponding compound in which R 4 represents amino with formic acid; (m) producing a compound of formula I in which R 4 represents NHCO-alkyl, by reaction of a corresponding 5 compound in which R 4 represents amino with an alkanoic anhydride; (n) producing a compound of formula I in which R 6 represents (S)-OH, by elimination of a leaving group from a correpsonding compound in which R 6 represents the leaving 1Q group; (o) producing a compound of formula I in which R 6 represents H and R 5 represents 0, by elimination of a leaving group from a correpsonding compound in which R 6 represents the leaving group; 25 (P) producing a compound of formula I in which R 6 and R 7 together represent a second bond between the carbon atoms to which they are attached, by elimination of a leaving group from a correpsonding compound in which R 6 represents the leaving group; 20 (9) producing a compound of formula I in which Y represents a cyclic group of formula III and R 8 represents CHO, by elimination of a leaving group from a correpsonding compound in which R 6 represents the leaving group; ♦ 6 25 (r) producing a compound of formula I in which R represents halogen, by reaction of a corresponding compound in which R 6 represents a leaving group with halide ion; (s) producing a compound of formula I in which R 5 and R 8 together represent a second bond between the carbon atoms to which they are attached, by elimination of halogen and alkoxy from a corresponding compound in which R 5 represents alkoxy and R 8 represents halogen; 5 (t) producing a compound of formula I in which R 5 represents (H,H) and R 8 represents H, by reduction of a corresponding compound in which R 5 and R 8 together represent a second bond between the carbon atoms to which they are attached; Ιθ (u) producing a compound of formula I in which R 8 represents H, by the action of hydride on a corresponding compound in which R 8 represents a leaving group; (v) producing a compound of formula I in which R 8 represents amino, by reduction of a corresponding compound 15 in which R 8 represents azido; (w) producing a compound of formula I in which R 8 represents alkylamino or alkanoylamino, by reaction of a corresponding compound in which R 8 represents amino with a suitable alkylating or acylating reagent; 20 (x) producing a compound of formula I in which R 8 represents alkyl substituted by OH, by reduction of a corresponding compound in which R 8 represents alkyl substituted by =0; (y) producing a compound of formula I in which R 8 25 includes a carboxylic acid group, by oxidation of a corresponding compound in which R 8 includes an aldehyde group; or (z) producing a compound of formula I in which R 8 represents optionally substituted alkenyl, by a Wittig reaction between a corresponding compound in which R 8 includes an aldehyde and an appropriate Wittig reagent.
13. A compound of formula I given and defined in claim 1 or a pharmaceutically acceptable derivative thereof, substantially as hereinbefore described and exemplified.
14. A process for the production of a compound of formula I given and defined in claim 1 or a pharmaceutically acceptable derivative thereof, substantially as hereinbefore described and exemplified.
15. A compound of formula I given and defined in claim 1 or a pharmaceutically acceptable derivative thereof, whenever produced by a process claimed in claim 12 or 14.
16. Use according to claim 8, substantially as hereinbefore described.
17. A pharmaceutical composition according to claim 9, substantially as hereinbefore described.
18. Use according to claim 10, substantially as hereinbefore described.
IE084791A 1990-03-13 1991-03-13 Immunosuppressive macrocyclic compounds IE910847A1 (en)

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GB909009480A GB9009480D0 (en) 1990-04-27 1990-04-27 Compounds
GB909017447A GB9017447D0 (en) 1990-08-09 1990-08-09 Compounds
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