HK1031731B - Epothilone derivatives - Google Patents

Description

Epothilone derivatives

Technical Field

The present invention relates to Epothilone derivatives, to processes for the preparation thereof and to intermediates thereto.

Background

Epothilones (epothilones) are macrolide compounds that are useful in the pharmaceutical field. For example, epothilones (epothilones) a and B have the following structures:

i epothilone A R ═ H

II epothilone B R ═ Me

It has now been found to have a similar microtubule-stabilizing effect as paclitaxel and thus to be cytotoxic to rapidly proliferating cells, e.g. tumour cells or other hyperproliferative cell diseases, see angelw.chem.int.ed.engl. (applied chemistry, international english edition), 1996, 35, No. 13/14.

Summary of The Invention

The present invention relates to compounds of the formula and any salt, solvate or hydrate thereof

Q is selected from

G is selected from the group consisting of alkyl, substituted or unsubstituted aryl, heterocyclic,

w is O or NR₁₅；

X is O or H, H;

y is selected from O; h, OR₁₆；OR₁₇，OR₁₇；NOR₁₈；H，NOR₁₉；H，NR₂₀R₂₁(ii) a H, H; or CHR₂₂；

OR₁₇OR₁₇Can be cyclic ketal;

Z₁and Z₂Is selected from CH₂，O，NR₂₃S, or SO₂Wherein Z is₁And Z₂Only one of which may be a heteroatom;

B₁and B₂Selected from OR₂₄Or OCOR₂₅Or O₂CNR₂₆R₂₇(ii) a When B is present₁When H and Y are OH, H, they may form six-membered cyclic ketals or acetals;

d is selected from NR₂₈R₂₉，NR₃₀COR₃₁Or a saturated heterocycle;

R₁，R₂，R₃，R₄，R₅，R₆，R₇，R₁₃，R₁₄，R₁₈，R₁₉，R₂₀，R₂₁，R₂₂，R₂₆and R₂₇Selected from H, alkyl, substituted alkyl or aryl, and when R is₁And R₂When alkyl, it may participate in the formation of cycloalkyl; when R is₃And R₄When alkyl, it may also participate in the formation of cycloalkyl;

R₉，R₁₀，R₁₆，R₁₇，R₂₄，R₂₅and R₃₁Selected from H, alkyl, or substituted alkyl;

R₈，R₁₁，R₁₂，R₂₈，R₃₀，R₃₂，R₃₃and R₃₀Selected from H, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, or heterocyclyl;

R₁₅，R₂₃and R₂₉Selected from H, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, heterocyclyl, R₃₂C＝O，R₃₃SO₂Hydroxyl, O-alkyl or O-substituted alkyl.

With the proviso that

The invention does not include

W and X are both O; and

R₁，R₂，R₇is H; and

R₃，R₄，R₆is methyl; and

R₈is H or methyl; and

Z₁and Z₂Is CH₂(ii) a And

g is 1-methyl-2- (substituted-4-thiazolyl) ethenyl;

q is as defined above

A compound of formula (V).

Detailed Description

The following definitions of various terms are used to describe the present invention. Unless otherwise specifically limited, the following definitions of terms are used throughout this application, whether for individual groups or for a portion of a larger group.

The term "alkyl" refers to a straight or branched chain unsubstituted hydrocarbon group having 1 to 20 carbon atoms, preferably 1 to 7 carbon atoms. The term "lower alkyl" refers to unsubstituted alkyl groups having from 1 to 4 carbon atoms.

The term "substituted alkyl" refers to alkyl substituted with, for example, one to four substituents such as: halogen, trifluoromethyl, trifluoromethoxy, hydroxy, alkoxy, cycloalkoxy, heterocyclyloxy, oxo, alkanoyl, aryloxy, alkanoyloxy, amino, alkylamino, arylamino, aralkylamino, cycloalkylamino, heterocyclylamino, disubstituted amino wherein 2 amino substituents are selected from alkyl, aryl, or aralkyl, alkanoylamino, aroylamino,aralkylaminoamino, substituted alkanoylamino, substituted arylamino, substituted aralkylaminoamino, mercapto, alkylthio, arylthio, aralkylthio, cycloalkylthio, heterocyclylthio, alkylthiocarbonyl, arylthiocarbonyl, aralkylthiocarbonyl, alkylsulfonyl, arylsulfonyl, aralkylsulfonyl, sulfamoyl (e.g., SO)₂NH₂) Substituted sulfamoyl, nitro, cyano, carboxy, carbamoyl (e.g. CONH)₂) Substituted carbamoyl (e.g., CONH alkyl, CONH aryl, CONH aralkyl, or two substituents on the nitrogen selected from alkyl, aryl or aralkyl), alkoxycarbonyl, aryl, substituted aryl, guanidino and heterocyclic groups such as indolyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidinyl, pyridyl, pyrimidinyl, and the like. It is to be noted that the above-mentioned substituents may be further substituted with halogen, alkyl, alkoxy, aryl or aralkyl.

The term "halogen" or "halo" refers to fluorine, chlorine, bromine and iodine.

The term "aryl" refers to monocyclic or bicyclic aromatic hydrocarbon groups having 6 to 12 carbon atoms in the ring, such as phenyl, naphthyl, biphenyl and biphenyl groups, each of which may be substituted.

The term "aralkyl" refers to an aryl group directly bonded to an alkyl group, such as benzyl.

The term "substituted aryl" refers to aryl substituted with, for example, one to four substituents such as: alkyl, substituted alkyl, halogen, trifluoromethoxy, trifluoromethyl, hydroxy, alkoxy, cycloalkoxy, heterocyclyloxy, alkanoyl, alkanoyloxy, amino, alkylamino, aralkylamino, cycloalkylamino, heterocyclylamino, dialkylamino, alkanoylamino, mercapto, alkylthio, cycloalkylthio, heterocyclylthio, ureido, nitro, cyano, carboxy, carboxyalkyl, carbamoyl, alkoxycarbonyl, alkylthiocarbonyl, arylthiocarbonyl, alkylsulfonyl, sulfamoyl, aryloxy and the like. The above substituents may be further substituted with halogen, hydroxy, alkyl, alkoxy, aryl, substituted alkyl or aralkyl.

The term "cycloalkyl" refers to an optionally substituted, saturated cyclic hydrocarbon ring system, preferably containing 1 to 3 rings, each ring having 3 to 7 carbon atoms, which may be further substituted with an unsaturated C₃-C₇The carbocyclic rings are fused. Examples of such groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclodecyl, cyclododecyl and adamantyl. Examples of the substituent include one or more of the above alkyl groups, or one or more of the above alkyl substituents.

The terms "heterocycle", "heterocyclic" and "heterocyclyl" refer to an optionally substituted, fully saturated or unsaturated, aromatic or nonaromatic cyclic group which is, for example, a 4 to 7 membered monocyclic, 7 to 11 membered bicyclic, or 10 to 15 membered tricyclic ring system which has at least one heteroatom in at least one carbon atom-containing ring. There may be 1, 2 or 3 heteroatoms selected from nitrogen, oxygen and sulfur in each heterocyclic ring containing a heteroatom, where the nitrogen and sulfur heteroatoms may also optionally be oxidized and the nitrogen heteroatoms may also optionally be quaternized. The heterocyclic group may be attached at any heteroatom or carbon atom.

Examples of monocyclic heterocyclic groups include pyrrolidinyl, pyrrolyl, indolyl, pyrazolyl, oxetanyl, pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl, oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl, thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl, tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxaazapyrrolidinylRadical, azaA group, 4-piperidonyl group, pyridyl group, N-oxopyridyl group, pyrazinyl group, pyrimidinyl group, pyridazinyl group, tetrahydropyranyl group, tetrahydrothiopyranyl sulfone group,morpholinyl, thiomorpholinyl sulfoxide, thiomorpholinyl sulfone, 1, 3-dioxopentyl and tetrahydro-1, 1-dioxothienyl, dioxacyclohexyl, isothiazolidinyl, thietanyl, triazinyl, triazolyl, and the like.

Examples of bicyclic heterocyclic groups include benzothiazolyl, benzoxazolyl, benzothienyl, quinuclidinyl, quinolinyl, quinolyl-N-oxide, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyranyl, indolizinyl, benzofuranyl, chromonyl, coumarinyl, 1, 2-naphthyridinyl, quinoxalinyl, indazolyl, pyrrolopyridyl, furopyridyl (e.g., furo [2, 3-c ] pyridyl, furo [3, 1-b ] pyridyl, or furo [2, 3-b ] pyridyl), isoindolyl, dihydroquinazolinyl (e.g., 3, 4-dihydro-4-oxoquinazolinyl), benzisothiazolyl, benzisoxazolyl, benzodiazinyl, benzofurazanyl, benzothioxanyl, benzotriazolyl, benzopyrazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl sulfone, dihydrobenzopyranyl, indolinyl, isochromonyl, isoindolinyl, 1, 5-naphthyridinyl, 2, 3-naphthyridinyl, piperonyl, purinyl, pyridopyridyl, quinazolinyl, tetrahydroquinolyl, thienofuranyl, thienopyridyl, thienothienyl, and the like.

Examples of substituents include one or more alkyl groups as described above or one or more alkyl substituents as described above. Also included are smaller heterocycles, such as epoxides and 1-aziridines.

The term "heteroatom" includes oxygen, sulfur and nitrogen.

The compounds of formula (V) may form salts with alkali metals such as sodium, potassium and lithium, with alkaline earth metals such as calcium and magnesium, with organic bases such as dicyclohexylamine, tributylamine, pyridine, and amino acids such as arginine, lysine, and the like. If the compound of formula (V) contains a carboxylic acid, the above salt can be obtained by exchanging the carboxylic acid protons for the desired ion, either in the medium in which the salt precipitates or in an aqueous medium followed by evaporation. Other salts can be prepared using techniques known to those skilled in the art.

The compounds of formula (V) may form salts with various organic and inorganic acids. Such salts include those formed with hydrochloric, hydrobromic, methanesulfonic, isethionic, sulfuric, acetic, trifluoroacetic, maleic, benzenesulfonic, toluenesulfonic acids and others (e.g., nitrates, phosphates, borates, tartrates, citrates, succinates, benzoates, ascorbates, salicylates, and the like). The salts may be obtained by reacting the compound of formula (V) with one equivalent of an acid in a medium in which the salt precipitates or in an aqueous medium which can be evaporated.

In addition, zwitterions ("inner salts") may be formed.

The compounds of formula (V) may also have prodrug forms. Any compound that can be converted in vivo to a biologically active agent (i.e., a compound of formula (V)) is a prodrug within the scope and spirit of the present invention.

For example, the compounds of formula (V) may form carboxylic acid esters. Carboxylic acid esters are conveniently obtained by esterifying the carboxylic acid functional groups present on the disclosed ring structures.

Various forms of prodrugs are known in the art. Examples of such prodrug derivatives can be found in:

a)design of Prodrugs (prodrug Design)Bundgaard eds, (Elsevier, 1985), andmethods in Enzymology (Methods in Enzymology),Vol.42p.309-396, K.Widder et al eds (Acamedic Press, 1985);

b)a Textbook of Drug Design and Development Development course)Eds by Krosgaard-Larsen and H.Burdgaard, Chapter 5, "design and Application of Prodrugs", H.Burdgaard, p.113-191 (1991);

c)H.Bundgaard，advanced Drug Delivery Reviews (Advanced drugs)Article (A) Release comments)，8，1-38(1992)；

d) H, Bundgaard, etc.,journal of Pharmaceutical Sciences Journal of science，77285 (1988); and

e) n. kakeya et al,chem Phar Bull (chemical and pharmaceutical Notification)，32，692(1984)。

Obviously, solvates (e.g. hydrates) of the compounds of formula (V) are also included within the scope of the present invention. Solvation processes are generally known techniques.

Use and utility

The compounds of formula (V) are microtubule stabilizing agents. Thus, they can be used to treat a variety of cancers or other dysplastic diseases, including (but not limited to) the following:

-cancers, including cancers of the bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; including squamous cell carcinoma;

-hematopoietic tumors of lymphocytic lineage, including leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphoblastoma, T-cell lymphoblastoma, hodgkin's (Hodgkins) lymphoblastoma, non-Hodgkins (Hodgkins) lymphoblastoma, hairy cell lymphomas and burkitts' lymphomas;

-hematopoietic tumors of myeloid lineage, including acute and chronic myeloid leukemia and promyelocytic leukemia;

-mesenchymally derived tumors, including fibrosarcoma and rhabdomyosarcoma;

-other tumors, including melanoma, seminoma, teratocarcinoma (tetratocancinoma), neuroblastoma and glioma;

tumors of the central and peripheral nervous system, including astrocytomas, neuroblastomas, gliomas and schwannomas;

-mesenchymally derived tumors including fibrosarcoma, rhabdomyosarcoma and osteosarcoma; and

other tumors, including melanoma, xeroderma pigmentosum, keratoacanthoma, seminoma, thyroid follicular cancer and teratoma (teratocarcinoma).

The compounds of formula (V) may also inhibit tumor angiogenesis and thus may affect abnormal cell proliferation. The anti-angiogenic properties of the compounds of formula (V) are also useful in the treatment of various forms of blindness associated with retinal vascularization, arthritis, in particular inflammatory arthritis, multiple sclerosis, restenosis and psoriasis.

The compounds of formula (V) can initiate or inhibit apoptosis, a physiological cell death process important for normal development and homeostasis. Changes in the pathways of apoptosis are strongly associated with the pathogenesis of various diseases in humans. As modulators of apoptosis, the compounds of formula (V) are useful in the treatment of a variety of human diseases accompanied by aberrant apoptosis including cancer (particularly, but not limited to, follicular lymphoma, cancers associated with mutations in the p53 gene, hormone-related tumors of the breast, prostate and ovary, and precancerous lesions such as familial adenomatous polyposis), viral infections (including, but not limited to, herpes viruses, poxviruses, Epstein-Barr virus, Sindbis virus and adenoviruses), autoimmune diseases (including, but not limited to, systemic lupus erythematosus, immune-mediated glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory bowel disease and autoimmune diabetes), neurodegenerative diseases (including, but not limited to, Alzheimer's disease, AIDS-related dementia, Parkinson's disease, amyotrophic lateral sclerosis, retinitis pigmentation, spinal muscular atrophy and cerebellar degeneration), aids, myelodysplastic syndrome, aplastic anemia, ischemic injury associated with myocardial infarction, stroke and reperfusion injury, cardiac arrhythmia, atherosclerosis, toxin-induced or alcohol-induced liver disease, hematologic disorders (including but not limited to chronic anemia and aplastic anemia), degeneration of the musculoskeletal system (including but not limited to osteoporosis and arthritis), aspirin-sensitive sinusitis, cystic fibrosis, multiple sclerosis, nephropathy, and cancer pain.

The compounds of the present invention may also be used in combination with known anti-cancer and cytotoxic drugs and treatments, including radiation therapy. If a fixed dose formulation is formulated, the combination product employs the compound of the invention in the dosage range described below and other pharmaceutically active agents within the allowable dosage range. When the conjugate formulation is not appropriate, the compound of formula (V) may be used sequentially with known anti-cancer or cytotoxic drugs and treatments. Particularly useful are cytotoxic drug binding agents wherein the compound of formula (V) is at G₂The M phase acts, while the second drug acts at a different phase of the cell cycle, e.g.the S phase.

For example thymidylate synthase inhibitors

DNA crosslinking agent

Topoisomerase I and II inhibitors

DNA alkylating agent

Nucleoside reductase inhibitors

Cytotoxic factors, e.g. TNF-alpha, or

Growth factor inhibitors, e.g. HER 2 receptor MAB

The compounds of the present invention may exist in a variety of optical, geometric and stereoisomeric forms. All such isomers and mixtures thereof are included within the scope of the present invention.

The compounds of the present invention may be formulated with pharmaceutically acceptable carriers or diluents for oral, intravenous or subcutaneous administration. Pharmaceutical compositions may be formulated in accordance with standard procedures using solid or liquid carriers, diluents and additives appropriate to the mode of administration desired. For oral formulations, the compounds of the present invention may be administered in the form of tablets, capsules, granules, powders, and the like. The compounds of the invention may be administered in a dosage range of about 0.05 to 200 mg/kg/day, preferably less than 100 mg/kg/day, in single dose form or in 2 to 4 divided doses.

Preferred compounds

Particularly preferred compounds of the formula (V)

Or

X is O

Y is O

Z₁And Z₂Is CH₂And an

W is NR₁₅。

Preparation method

The compounds of formula (V) may be prepared according to the following scheme.

Scheme 1

V (Q is vinyl) V (Q is ethylene oxide)

Wherein R is₃，R₄，R₅，R₆，R₈And R₁₅As defined above, P₁Is an oxygen protecting group.

Wherein W is NR₁₅And X is O can be prepared as described in scheme 1. Wherein P is₁Compounds of formula XII, which are oxygen protecting groups such as t-butyldimethylsilyl, can be synthesized in accordance with known procedures (i.e., Nicolaou, k.c., etc.,angew.chem.int.ed.engl. (applied chemistry, international English edition)(1997)36, 166-168) from a compound of formula VI. The aldol condensation of the compound of formula XII and the compound of formula XIV gives the compound of formula XIII. The compounds of formula XIV can be prepared by known methods (i.e., Schinzer, d. et al,eur.chem.chron. (european chemistry) Note)(1996)1, 7-10). The aldehyde of formula XVIII can be synthesized as shown in scheme 1 or by using known methods (i.e., Taylor, r.e., etc.,Tetrahedron lett.(tetrahedron letters), (1997), 38, 2061-2064) were prepared from compounds of formula XV. Compounds of formula XIX can be prepared by treating compound XVIII with an amine using dehydration conditions such as catalytic amounts of p-toluenesulfonic acid and azeotropic removal of water. The compound of formula XX may be prepared by treating the compound of formula XIX with an allylating reagent such as allylmagnesium bromide. Compounds of formula XXI can be prepared from compounds of formula XIII and XX by using standard amide bond coupling reagents (i.e., DCC, BOP, EDC/HOBT, PyBrOP). By using Grubbs (RuCl)₂(＝CHPh)(PCY₃)₂(ii) a See Grubbs, r.h. et al, angelw.chem.int.ed.engl. (applied chemistry, international english edition), (1995)34, 2039) or Schrock catalysts (see Schrock, R.R et al,am chem soc (american society of chemistry)(1990)112, 3875) to carry out ring-closing metathesis, a compound of the formula XXII can be prepared from a compound of the formula XXI. For example, when P₁Deprotection of a compound of formula XXI, wherein Q is vinyl and W is NR, to give a compound of formula XXI wherein Q is vinyl and W is NR, using hydrogen fluoride in acetonitrile or tetra-n-butylammonium fluoride in THF in the case of t-butyldimethylsilyl₁₅X is O and R₃、R₄、R₅、R₆The compound of formula V as defined above. Regioselective epoxidation of compounds of formula V wherein Q is vinyl using dimethyldiepoxide to give compounds wherein Q is oxirane and W is NR₁₅X is O and R₃、R₄、R₅、R₁₅The compound of formula V as defined above.

Scheme 2

Alternatively, as shown in scheme 2, by reacting a compound of formula XXIII with magnesium and an acid chloride (R)₅CH₂COCl) to give a compound of formula XXIV (see, e.g., Heathcock, c., etc.,chem. (journal of organic chemistry)1990, 55, 1114) -1117) followed by ozonolysis to give the compound of formula VIII.

Scheme 3

Alternatively, compounds of formula XIV can be prepared as shown in scheme 3. Reaction of a compound of formula XXV with pseudoephedrine affords a compound of formula XXVI. By applying the method of Meyers (i.e., Meyers, a. etc.,am chem soc (american society of chemistry)1994, 116, 9361-9362), compounds of the formula XXVII may be prepared by alkylation of compounds of the formula XXVI with pentenyl halides, for example 5-bromopentene. Compounds of formula XXVIII can be prepared from compounds of formula XXVII using a reducing agent such as lithium pyrrolidinoborohydride. Oxidation of a compound of formula XXVIII using, for example, pyridinium chlorochromate affords a compound of formula XIV. The compound of formula XXVII can be directly converted to the compound of formula XIV using a reducing agent such as lithium aluminum triethoxy hydride.

Scheme 4

Alternatively, compounds of formula XX can be prepared from allylglycine according to scheme 4. According to the field of forceN-protection of allylglycine by methods known in the art gives compounds of formula XXIX, wherein P₂Suitable groups for N-protection are, for example, tert-butoxycarbonyl. Optionally, when R₂₉Compounds of formula XXX may be prepared by alkylation of compounds of formula XXIX with an alkyl halide in the presence of a base such as sodium hydride, when other than hydrogen. Compounds of formula XXXI can be prepared from compounds of formula XXX using N, O-dimethylhydroxylamine and standard coupling reagents such as EDCI and HOBT. Compounds of formula XXXII can be prepared from hydroxamate XXXI by treatment with organometallic reagents such as alkyl or aryl magnesium halides. Wittig olefination of a compound of formula XXXII affords a compound of formula XXXIII (Wittig reagent as Danishefsky, S.E. et al,chem. (journal of organic chemistry)1996, 61, 7998-. N-deprotection of a compound of formula XXXIII by methods known in the art affords compounds of formula XX.

Scheme 5

V (Q is vinyl) V (Q is ethylene oxide)

Wherein W is NR can be prepared as shown in scheme 5₁₅Compounds of formula V wherein X is oxygen and G is a 1, 2-disubstituted alkene. Compounds of formula XXXV can be obtained by Wittig olefination of compounds of formula XXXII. Compounds of formula XXXIV may be prepared according to methods known in the art. Compounds of formula XXXVI may be prepared by N-deprotection of compounds of formula XXXV according to methods known in the art. Compounds of formula XXXVII may be prepared by coupling compounds of formula XXXVI and XIII using standard coupling reagents such as EDCI and HOBT. Following the procedure for the preparation of compounds of formula XXII in scheme 1, compounds of formula XXXVIII may be prepared from compounds of formula XXXVII. Compounds of formula XXXVIII may be converted to compounds wherein W is NR by the methods described in scheme 1 (steps o and p)₁₅Compounds of formula V in which X is oxygen and G is a 1, 2-disubstituted alkene。

Scheme 6

XXXXXII V (Q is vinyl) V (Q is ethylene oxide)

Compounds of formula V wherein W and X are oxygen and G is a 1, 2-disubstituted alkene can be prepared as shown in scheme 6. The compounds of formula XXXX may be prepared by treating a compound of formula XXXIX with an allylating reagent such as allylic magnesium bromide. Enantiomerically pure compounds of formula XXXX can be prepared using chiral reagents (see, e.g., Taylor, r.e., et al,tetrahedron Let t (Tetrahedron bulletin)1997, 38, 2061-2064; nicolaou, k.c., etc.,Angew.Chem.Int.Ed.Engl. (applied chemistry, International English edition)1997, 36, 166-; keck, G, et al,am chem soc (american society of chemistry),1993, 115, 8467). Compounds of formula XXXXI can be prepared from compounds of formulae XXXX and XIII by using standard esterification agents such as DCC and DMAP. Compounds of formula XXXXII can be prepared from formula XXXXI by olefin ring-closing metathesis as described in scheme 1 for compounds of formula XXXII. Compounds of formula V wherein W and X are oxygen and G is a 1, 2-disubstituted alkene can be prepared from compounds of formula XXXXII by deprotection (when Q is vinyl) and, if desired, epoxidation as described above (when Q is oxiranyl).

Scheme 7

XXXXXXVI V (Q is vinyl) V (Q is ethylene oxide)

Compounds of formula V wherein W and X are oxygen and G is alkyl, substituted alkyl, aryl, heteroaryl, bicyclic aryl or bicyclic heteroaryl can be prepared as shown in scheme 7. Compounds of formula XXXXIV may be prepared by allylating a compound of formula XXXIII wherein G is alkyl, substituted alkyl, aryl, heteroaryl, bicyclic aryl or bicyclic heteroaryl by reaction with an allylating reagent such as allylmagnesium bromide. The compound of formula XXXXV can be prepared by esterification of a compound of formula XIII with a compound of formula XXXXIV using, for example, DCC and DMAP. Compounds of formula xxxxxvi can be prepared from compounds of formula XXXXV by ring-closing metathesis as described above. Compounds of formula XXXXVI may be converted to compounds of formula V by deprotection followed by epoxidation as described in scheme 1.

Scheme 8

L V (Q is vinyl) V (Q is ethylene oxide)

Where W is NR can be prepared as shown in scheme 8₁₅A compound of formula V wherein X is oxygen and G is alkyl, substituted alkyl, aryl, heteroaryl, bicyclic aryl, or bicyclic heteroaryl. Compounds of formula XXXXVI are prepared by reacting a compound of formula XXXIII wherein G is alkyl, substituted alkyl, aryl, heteroaryl, bicyclic aryl or bicyclic heteroaryl with an amine under dehydrating conditions. Compounds of formula XXXXXVIII may be prepared from compounds of formula XXXXXVII by treatment with an allylating reagent such as allylmagnesium bromide. Compounds of formula XXXXIX can be prepared from compounds of formula XXXXVIII and compounds of formula XIII by using standard amide bond coupling reagents, e.g., EDCI and HOBT. Compounds of formula L can be prepared from compounds of formula XXXXIX by ring-closing metathesis as described above. According to the method described in scheme 1, by removingProtection and subsequent epoxidation, the compound of formula L can be converted to the compound of formula V.

Scheme 9

Where W is NR can be prepared as shown in scheme 9₁₅X is oxygen, G is

And D is selected from NR₂₈R₂₉，NR₃₀COR₃₁And standard heterocycles (i.e., piperidinyl, morpholinyl, piperazinyl, etc.) of formula V. Compounds of formula LI may be prepared by reductive amination of compounds of formula XXXII using a primary or secondary amine and a reducing agent such as sodium triacetoxyborohydride. Compounds of formula LIII, LIV, and V can then be prepared as described in scheme 1.

Scheme 10

Alternatively, compounds of formula V wherein X is oxygen, W is oxygen or NR can be prepared as shown in scheme 10₁₅Or oxygen, G is

And D is selected from NR₂₈R₂₉，NR₃₀COR₃₁And standard heterocycles (i.e., piperidines)Phenyl, morpholinyl, piperazinyl, etc.). The compound of formula V may be converted to the compound of formula LV by protecting the hydroxy group with a suitable protecting group such as tert-butyldimethylsilyl. Compounds of formula LVI can be prepared from compounds of formula LV by ozonolysis. Treatment of a compound of formula LVI with an amine and a reducing agent such as sodium triacetoxyborohydride affords a compound of formula LVII. Removal of the protecting group of the compound of formula LVII with, for example, hydrogen fluoride, gives a compound wherein X is oxygen and W is NR₁₅Or is oxygen, G is

A compound of formula V.

Scheme 11

Wherein W is NR can be prepared as shown in scheme 11₁₅X is oxygen, G is

A compound of formula V. Compounds of formula LVIII can be prepared from compounds of formula XXX by treatment with an amine and standard amide bond coupling reagents such as EDCI and HOBT. When P is present₂In the case of t-butoxycarbonyl, the compound of formula LX can be prepared from the compound of formula LVIII by N-deprotection using, for example, trifluoroacetic acid, followed by coupling of the compounds of formulae LIX and XIII using standard amide bond coupling reagents such as EDCI and HOBT. LXI compounds can be prepared from compounds of formula LX by ring-closing metathesis. Compounds of formula V can be prepared from compounds of formula LXI according to the procedures described in scheme 1.

Scheme 12

According to scheme 12 wherein W is oxygen, X is oxygen, and G is

A compound of formula V. The compound of formula LXII can be prepared by treating allyl glycine with nitrous acid. Compounds of formula LXIII can be prepared from compounds of formula LXII by treatment with an amine and standard amide bond coupling reagents such as EDCI and HOBT. Compounds of formula LXIV can be prepared from compounds of formulae LXIII and XIII using standard amide bond coupling reagents such as EDCI and HOBT. LXV compounds can be prepared from compounds of formula LXIV by ring-closing metathesis. The compound of formula V can be prepared from the compound of formula LXV according to the procedures described in scheme 1.

Scheme 13

As shown in scheme 13, compounds of formula V wherein G is a 1, 2-disubstituted ethyl group can be prepared by using a catalyst such as palladium on hydrocarbon compounds of formula V wherein G is a 1, 2-disubstituted vinyl group. In addition, compounds of formula V wherein G is a 1, 2-disubstituted cyclopropyl group can be prepared from compounds of formula V wherein G is a 1, 2-disubstituted vinyl group by cyclopropanation using diiodomethane and a zinc-copper coupling agent as described in scheme 4.

Scheme 14

According to scheme 14, wherein Z can be prepared₁A compound of formula V which is oxygen. Compounds of formula LXVII can be prepared from the alpha-hydroxy ester LXVI by reaction with 3-buten-1-yl-trifluoromethanesulfonate (or with 3-butenyl bromide and silver triflate (silver triflate)). The compound of formula LXVII can be reduced with a reducing agent such as diisobutylaluminum hydride to give the compound of formula LXVIII. Alternatively, compounds of formula LXVIII can be obtained from compounds of formula LXVII by a two-step reaction comprising reduction with lithium borohydride and oxidation with pyridinium chlorochromate. A compound of formula LXVIII can be substituted for the compound of formula XIV in scheme 1 to provide a compound of formula LXIX. Further reaction of LXIX as described above to give Z therein₁A compound of formula V which is oxygen.

Scheme 15

Similarly, as shown in scheme 15, wherein Z can be prepared₁Is NR₂₃A compound of formula V. Compounds of formula LXXI can be prepared by reacting an alpha-amino ester LXX with 3-buten-1-yl bromide. The compound of formula LXXI can be reduced with a reducing agent such as diisobutylaluminum hydride to give the compound of formula LXXII. Alternatively, compounds of formula LXXII can be obtained from compounds of formula LXXI by a two-step reaction comprising reduction with lithium borohydride and oxidation with pyridinium chlorochromate. A compound of formula LXXII can be substituted for the compound of formula XIV in scheme 1 to provide a compound of formula LXXII, which is further reacted as described above to provide wherein Z₁Is NR₂₃A compound of formula V.

Scheme 16

According to scheme 16, wherein Z can be prepared₂A compound of formula V which is oxygen. The compounds of formula LXXV can be prepared from the β -hydroxy ester LXXIV with an allylating agent such as allyl bromide (or with allyl bromide and silver triflate). The compound of formula LXXV can be reduced with a reducing agent such as diisobutylaluminum hydride to give the compound of formula LXXVI. Alternatively, compounds of formula LXXVI can be obtained from compounds of formula LXXV by a two-step reaction comprising reduction with lithium borohydride and oxidation with pyridinium chlorochromate. A compound of formula LXXVI can be substituted for compound of formula XIV in scheme 1 to give a compound of formula LXXVII. Further reaction of LXXVI as described above gives the compound in which Z₂A compound of formula V which is oxygen.

Scheme 17

Similarly, as shown in scheme 17, wherein Z can be prepared₂Is NR₂₃A compound of formula V. The compounds of formula LXXIX can be prepared from the β -amino ester LXXVIII with an allylating reagent such as allyl bromide. The compound of formula LXXIX can be reduced with a reducing agent such as diisobutylaluminum hydride to give the compound of formula LXXX. Alternatively, compounds of formula LXXX can be obtained from compounds of formula LXXIX by a two-step reaction comprising reduction with lithium borohydride and oxidation with pyridinium chlorochromate. Compounds of formula LXXX can be substituted for compounds of formula XIV in scheme 1 to provide compounds of formula LXXXI. Further reaction of LXXXI as described above gives the compound in which Z₂Is NR₂₃A compound of formula V.

Scheme 18

According to scheme 18 wherein W isOxygen or NR₁₅And Y is H, a compound of formula V of H. Compounds of formula V can be converted to compounds in which P is present by treatment with a reagent such as (tert-butyldimethylsilyl) trifluoromethanesulfonate₄And P₅A compound of formula LXXXII which is a hydroxy protecting group. Compounds of formula LXXXII can be prepared by treating a compound of formula LXXXII with Lawesson's reagent. By reduction with a reducing agent, e.g. tri-n-butyltin hydride, when W is oxygen, or by NR₁₅Compounds of formula LXXXIV can be prepared from compounds of formula LXXXIII by treatment with methyl iodide and sodium borohydride. When P is present₄And P₅In the case of a silyl group, the protecting group of the compound of formula LXXXIV is removed, for example, with hydrogen fluoride, to give a compound in which W is oxygen or NR₁₅And Y is H, a compound of formula V of H.

Scheme 19

Wherein W and Y are oxygen and R is prepared as shown in scheme 19₁A compound of formula V which is alkyl or substituted alkyl. Protection of the compound of formula V by treatment with a reagent such as (tert-butyldimethylsilyl) trifluoromethanesulfonate may give a compound in which P is₅And P₆A compound of formula LXXXV which is a hydroxy protecting group. Compounds of formula LXXXVI can be prepared from compounds of formula LXXXV by treatment with a reducing agent such as sodium borohydride. Compounds of formula LXXXVI can be prepared by protecting the hydroxy group with P-methoxybenzyl trichloroacetimidate, wherein P₇A compound of formula LXXXVII which is p-methoxybenzyl. When P is present₅And P₆In the case of tert-butyldimethylsilyl, the protecting group P of the compound of formula LXXXVII is removed in pyridine using, for example, hydrogen fluoride₅And P₆A compound of formula LXXXXXVIII may be obtained which is then selectively protected using, for example, tert-butyldimethylsilyl chloride, to give a compound wherein P is₈A compound of formula LXXXXIX which is tert-butyldimethylsilyl. From compounds of formula LXXXIX by treatment with a base such as lithium diisopropylamide followed by treatment with an alkylating agent such as methyl iodidePreparing the compound of formula C. Compounds of formula C may be protected by treatment with a reagent such as (tert-butyldimethylsilyl) trifluoromethanesulfonate to give compounds in which P₉A compound of formula CI that is a hydroxy protecting group. When P is present₇For P-methoxybenzyl, P is removed by using, for example, DDQ₇Groups the compounds of formula CII can be prepared from compounds of formula CI. When P is present₈And P₉In the case of silyl groups, compounds of formula CII wherein W and Y are oxygen and R is hydrogen can be prepared from compounds of formula CII by oxidation using, for example, TPAP/NMO, followed by removal of the protecting group using, for example, hydrogen fluoride₁A compound of formula V which is alkyl or substituted alkyl. The compound of formula V above can be further oxidized using dimethyldioxirane as described in scheme 1 to give the corresponding epoxy compound of formula V.

Scheme 20

V (Q is ethylene oxide) V (Q is vinyl)

As shown in scheme 20, compounds of formula V wherein X is oxygen and Q is an alkene can be prepared from compounds of formula V wherein X is oxygen and Q is an oxirane ring by treatment with a reactive metallocene such as a titanocene, zirconocene, or niocene (see, e.g., r.schobert and u.hohlein,Synlett(synthetic Rapid newspaper) (1990), 465-466).

Scheme 21

As shown in scheme 21, compounds of formula V wherein X is oxygen and W is NR can be prepared from compounds of formula V wherein X and W are both oxygen₁₅Wherein R is₁₅Being hydrogenA compound of formula V. Compounds of formula CIII can be prepared from compounds of formula V wherein X and W are both oxygen by forming an allylpalladium π complex using a pendant such as tetrakis (triphenylphosphine) palladium, followed by treatment with sodium azide (see, e.g., Murahashi, S. -I, et al J.Org.Chem. (J.Org.chem.) 1989, 54, 3292). The compound of formula CIII is then reduced with a reducing agent such as triphenylphosphine to give the compound of formula CIV. Compounds of formula CIV wherein X is oxygen and W is NR can be prepared from compounds of formula CIV by macrocyclic lactamization using diphenylphosphorylazide or bromotripyrrolidinylphosphonium hexafluorophosphate (PyBroP)₁₅(wherein R is₁₅Hydrogen) to the compound of formula V.

Process 22

As shown in scheme 22, compounds of formula V wherein X is oxygen and W is NR can be prepared from compounds of formula V wherein X and W are both oxygen₁₅Wherein R is₁₅Compounds of formula V which are alkyl, substituted alkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, O-alkyl, O-substituted alkyl. Compounds of formula CV can be prepared from compounds of formula V wherein X and W are both oxygen by forming an allylpalladium π complex using, for example, tetrakis (triphenylphosphine) palladium, followed by treatment with a primary amine. Compounds of formula V wherein X is oxygen and W is NR can be prepared from compounds of formula V by macrocyclic lactamization using diphenylphosphorylazide or bromotripyrrolidinylphosphonium hexafluorophosphate (PyBroP)₁₅Wherein R is₁₅Compounds of formula V which are alkyl, substituted alkyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, hydroxy, O-alkyl, O-substituted alkyl. When R is₁₅In the case of OH, it may be desirable to remove R therefrom₁₅Removal of protecting groups such as t-butyldimethylsilyl groups for intermediates such as O-t-butyldimethylsilyl groups.

The in vitro assay for the biological activity of the compounds of formula (V) is carried out as follows:

tubulin in vitro polymerization

The method of Williams and Lee (see Williams, R.C., Jr. and Lee, J.C. preparation of tubulin from brain. methods in Enzymology (tubulin from brain, methods in Enzymology) was followed85And Pt.D: 376. sup. 385, 1982) second cycle (2X) calf brain tubulin was prepared and stored in liquid nitrogen prior to use. Tubulin polymerization was quantitatively analyzed as modified by Swindell et al (see Swindell, C.S., Krauss, N.E., Horwitz, S.B., and ringer, I.Biologic active taxol analogs with deleted A-ring side chain substituents and variable C-2' ligands.) J.Med.Chem.34: 1176-1184, 1991). These improvements can be summarized, to some extent, as the tubulin polymerization capacity expressed in terms of the effective concentration of a given compound. In this method, polymerization buffer (0.1M MES, 1mM EGTA, 0.5mM MgCl) was added to a microculture cell of a Beckman (Beckman Instrument) Model DU 7400 UV spectrophotometer at 37 ℃₂Ph6.6) was added to the polymerization buffer containing tubulin. The final tubulin concentrations typically used were 1.0mg/ml and the compound concentrations were 2.5, 5.0 and 10. mu.M. After manually determining the initial and final times of the linear region including at least 3 time points, the initial slope of the change in OD measured every 10 seconds was calculated by the program attached to the instrument. Under these conditions, the linearity difference is generally < 10^-6The slope ranged from 0.03 to 0.002 absorbance units/minute with a maximum absorbance of 0.15 absorbance units. Effective Concentration (EC)_0.01) The insert concentration, defined as the initial slope that results in a 0.01 OD/min ratio, was calculated using the following formula: EC (EC)_0.01Concentration/slope. EC (EC)_0.01Values are expressed as mean and standard deviation values from 3 different concentrations. EC of the Compounds of the invention_0.01The value ranges from 0.01 to 1000. mu.M.

Cytotoxicity (in vitro)

Reported by T.L.Riss et al ("Compa)rison of MTT, XTT, and a noveltrazolium compound MTS for in vitro proliferative chemosensory assays, "(comparison of in vitro proliferation and chemosensitivity assays for MTT, XTT and a novel tetrazolium compound MTS) mol. 184a, 1992) cytotoxicity was evaluated in HCT-116 human colon carcinoma cells by MTS (3- (4, 5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfinyl) -2H-tetrazolium, inner salt) assay. Cells were seeded in 96-well microtiter plates at 4,000 cells/well and 24 hours later, drug was added and serial dilutions were performed. The cells were incubated at 37 ℃ for 72 hours at which time 333. mu.g/ml (final concentration) of the tetrazolium dye MTS bound to 25. mu.M (final concentration) of the electron coupler phenazine methosulfate was added. Dehydrogenases in living cells reduce MTS to a form that absorbs 492nM of light, which can be quantitatively determined by means of a spectrophotometer. The greater the absorbance, the greater the number of viable cells. IC for result₅₀Expressed as the drug concentration required to inhibit cell proliferation (i.e., absorbance at 450 nM) to 50% of that of untreated control cells. IC of the Compounds of the invention₅₀Values range from 0.01-1000 nM.

The following examples serve to illustrate the invention.

Example 1

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13 (E) -cyclohexadecene-2, 6-dione

N- [ (2-methyl) -1-propenyl ] morpholine

Morpholine (165.5g, 1.9mol) was added to isobutyraldehyde (173mL, 1.9mol) with stirring at a rate that did not cause the reaction temperature to exceed 30 ℃. After the addition was complete, the reaction mixture was stirred at room temperature for 2 hours and then heated at 160 ℃ for 20 hours in a flask equipped with a Dean-Stark trap. The reaction mixture was then cooled to room temperature and the flask was fitted with a vigreux distillation column. Distillation in vacuo gave 135g (50%) of Compound A as a clear, colorless oil. MS (M + H, 142).

B.2, 2-dimethyl-3-oxopentanal

A solution of propionyl chloride (44mL, 0.50mol) in diethyl ether (135mL) was added under a nitrogen atmosphere to a solution of Compound A (69g, 0.50mol) in diethyl ether (135mL) at 0 deg.C with stirring over 45 minutes. After the addition was complete, the reaction mixture was stirred at reflux for 2 hours and then at room temperature for 16 hours. The reaction mixture was filtered and the filter cake was washed with diethyl ether (50 mL). Volatiles were removed in vacuo. The residue was dissolved in water (80mL) and the solution was adjusted to pH 4. Ether (80mL) was added and the biphasic compound was stirred for 16 h. The reaction mixture was poured into a separatory funnel, and the aqueous layer was extracted with ether (5X 100 mL). The combined organic layers were dried (MgSO)₄) Filtered and evaporated in vacuo. The residue was distilled under high vacuum to give 10.4g (16%) of compound B as a clear colorless oil. MS (M-H, 127).

C.4-tert-butyldimethylsilyloxy-5, 5-dimethyl-6-oxo-1-octene

To a solution of (-) -B-methoxydiisopinocamphenylborane (25.7g, 81mmol) in diethyl ether (80mL) at 0 deg.C under nitrogen was added a 1.0M solution of allylmagnesium bromide in diethyl ether (77mL, 77mmol) over 1.5 hours. The reaction mixture was stirred at 25 ℃ for 1 hour and concentrated in vacuo. The residue was extracted with pentane (2X 150mL) and the extracts were filtered through Celite under a nitrogen atmosphere. The combined extracts were evaporated in vacuo to give B-allyldiisopinocamphenylborane. This material was dissolved in diethyl ether (200mL) under nitrogen and cooled to-100 ℃. Then, a solution of Compound B (11.42g, 89mmol) in diethyl ether (90mL) was added at-78 deg.C over 1 hour. The reaction mixture was stirred for a further 0.5 h and methanol (1.5mL) was added.The reaction mixture was warmed to room temperature and quenched with 3N NaOH (32mL) and 30% H₂O₂(64mL) and then refluxed for 2 hours. The reaction mixture was cooled to room temperature, the layers were separated and the organic phase was washed with water (500 mL). The combined aqueous phases were extracted with diethyl ether (2X 100 mL). The combined organic extracts were washed with saturated aqueous NaCl (100mL) and dried (MgSO)₄) Filtering and vacuum concentrating. The residue was dissolved in CH₂Cl₂(250mL), cooled to 0 ℃ and diisopropylethylamine (93mL, 535mmol) was added. Then, while stirring, trifluoromethanesulfonic acid (tert-butyldimethylsilyl) ester (69g, 260mmol) was slowly added at a rate such that the temperature did not exceed 10 ℃. After the addition was complete, the reaction mixture was poured into water (650mL), the layers were separated, and the aqueous layer was extracted with dichloromethane (2X 650 mL). The combined organic layers were dried (NaSO)₄) Filtering and vacuum concentrating. The residue was purified by flash chromatography, eluting first with hexane and then 10% ethyl acetate/hexane to give 17.2g (78%) of compound C as a clear colorless oil. By Mosher esters of alcohols¹The enantiomeric excess was 94% as determined by H NMR analysis.¹³C

NMR(CDCl₃，80MHz)d 215.8，136.1，116.5，52.8，39.0，31.9，26.0，22.4，20.1，18.1，7.6，-3.6，-4.4.

3-tert-Butyldimethylsiloxy-4, 4-dimethyl-5-oxoheptanal

O was bubbled through a solution of Compound C (10.8g, 38.0mmol) in dichloromethane at-78 deg.C₃Until the solution remained blue (1 hour). Then introducing O₂15 minutes after which N is introduced₂After 30 minutes, the solution finally became transparent. Triphenylphosphine (10g, 38mmol) was then added and the reaction mixture was warmed to-35 ℃ and left for 16 h. The volatiles were removed in vacuo and purified by flash chromatography, eluting with 8% ethyl acetate/hexanes to give 8.9g (74%) of compound D as a clear colorless oil.

¹H NMR(CDCl₃，300MHz)d 9.75(m，1H)，4.53(t，J＝4.8Hz，1H)，3.40-3.60(m，4H)，1.10(s，3H)，1.07(s，3H)，0.98(t，J＝7.0Hz，3H)，0.83(s，9H)，0.07(s，3H)，0.04(s，3H).

3-tert-Butyldimethylsiloxy-4, 4-dimethyl-5-oxoheptanoic acid

To a solution of compound D (3.90g, 13.6mmol) in t-butanol (75mL) was added 2-methyl-2-butene (5.85mL, 55.2mmol) at room temperature, followed by dropwise addition of a solution of sodium chlorite (4.61g, 40.8mmol) and sodium phosphate (2.81g, 20.4mmol) in water (15 mL). The reaction mixture was stirred for 0.5 h and then the solvent was removed in vacuo. Water (150mL) was added to the residue, followed by extraction with ethyl acetate (3X 150 mL). The combined organic extracts were dried (MgSO)₄) Filtered and the volatiles removed in vacuo. The residue was purified by flash chromatography, eluting with 20% ethyl acetate/hexane/1% acetic acid, to give 3.97g (92%) of compound E as a clear, colorless, viscous oil. MS (M + H, 303).

(R, R) -N- (2-hydroxy-1-methyl-2-phenylethyl) -N, 2- (S) -dimethyl-6-heptenamide

A suspension of LiCl (6.9g, 0.16mol) and preformed lithium diisopropylamide (Aldrich, 2.0M heptane/ethylbenzene/THF solution, 27.6mL, 55mmol) plus THF (70mL) was treated with dropwise addition of (R, R) -N- (2-hydroxy-1-methyl-2-phenylethyl) -N-methylpropionamide (6.0g, 27mmol, Meyers, A.G., et al J.am.chem.Soc. (Proc. chem.Chem.J.) in THF (30mL) at-78 deg.C over 10 minutes. The bright yellow reaction mixture was stirred at-78 deg.C for 1 hour, 0 deg.C for 15 minutes, 25 deg.C for 5 minutes, then cooled to 0 deg.C and treated with a solution of 5-bromo-1-pentene (4.8mL, 40mmol) in THF (5 mL). The reaction mixture was stirred at 0 ℃ for 24 hours and NH was poured in₄Saturated aqueous Cl (100mL) and ethyl acetate (100 mL). The phases were separated and the aqueous phase was extracted with further ethyl acetate (3X 100 mL). The organic extracts were combined, washed with saturated aqueous sodium chloride (200mL), and dried (Na)₂SO₄) And (4) concentrating in vacuum. Flash chromatography (SiO)₂，4.0×25cm，2％MeOH-CHCl₃) Compound F was obtained as a pale yellow oil (6.9g, 88%). MS (ESI)⁺)：290(M+H)⁺：MS(ESI^-)：288.2(M-H)^-。

(S) -2-methyl-6-heptenol

Pyrrolidine (2.6mL, 30mmol) and BH were added sequentially to a 250mL round bottom flask at 0 deg.C₃-THF complex (1.0M in THF, 31mL, 30 mmol). The borane-pyrrolidine complex was warmed to 25 deg.C (1 hour) and cooled to 0 deg.C, treated dropwise over 30 minutes with n-butyllithium (1.6M in hexane, 19mL, 30mmol) while carefully maintaining the internal temperature below 5.5 deg.C. The reaction mixture was stirred at 0 ℃ for another 30 minutes, and then a solution of compound F (3.0g, 10mmol) in THF (23mL) was added dropwise over 10 minutes. The reaction mixture was stirred (6 hours) at 25 ℃ and then quenched dropwise with 3N aqueous HCl (25 mL). The reaction mixture was then poured into 1N aqueous HCl (200mL) and extracted with ether (4X 80 mL). The combined organic phases were washed with 1: 1 saturated aqueous NaCl-1N aqueous HCl (2X 150mL) and concentrated in vacuo. To the residue was added aqueous NaOH (1N, 200mL) and the suspension was stirred for 30 min. The mixture was extracted with diethyl ether (3X 100mL), the combined diethyl ether layers were washed with 1: 1 saturated aqueous NaCl-1N aqueous NaOH (2X 200mL), and dried (Na)₂SO₄) And (4) concentrating in vacuum. By flash chromatography (SiO)₂4.0X 25cm, 15-25% diethyl ether-pentane gradient) to give mixture G (1.26G, 95%) as a colorless oil. [ a ] A]²⁵ _D-11(c 12，CH₂Cl₂)。

(S) -2-methyl-6-heptanal

Treatment of Compound G (0.24G, 1.9mmol) with pyridinium chlorochromate (0.61G, 2.8mmol) for CH₂Cl₂(6mL) and the reaction mixture was stirred at 25 ℃ for 5 hours. The resulting dark brown viscous paste was passed through a silica gel-Celite column (Celite 1.0X 1cm in SiO₂Over 1.0X 5cm, eluting with 50mL of dichloromethane). The solvent was removed in vacuo to give crude compound H (0.15g, 63%) as a colorless oil, which was pure enough for the subsequent reaction.

¹H NMR(300MHz，CD₂Cl₂)d 9.62(s，1H)，5.88-5.68(m，1H)，5.13-4.92(m，2H)，2.37-2.24(m，1H)，2.15-2.05(m，2H)，1.62-1.78(m，1H)，1.51-1.32(m，3H)，1.07(d，3H，J＝7.0Hz).

(3S, 6R, 7S, 8S) -3-tert-butyldimethylsilyloxy-4, 4, 6, 8-tetramethyl-7-hydroxy-5-oxo-12-tridecenoic acid

A solution of compound E (1.0g, 3.4mmol) in THF (5mL) was added dropwise over 3 minutes to a pre-formed LDA solution (Aldrich, 2.0M heptane/ethylbenzene/THF solution, 3.8mL, 7.6mmol) with additional THF (25mL) at-78 deg.C. The reaction mixture was stirred at-78 deg.C (10 min), warmed to-40 deg.C (20 min), and cooled to-78 deg.C, then compound H (0.56g, 4.4mmol) in THF (5mL) was added. The reaction mixture was warmed to-40 ℃, stirred for 1 hour, and diluted with a saturated aqueous solution of ammonium chloride (50 mL). The layers were separated and the aqueous layer was extracted with ethyl acetate (4X 50 mL). The combined organic layers were washed with saturated aqueous sodium chloride (100mL), dried (sodium sulfate) and concentrated in vacuo. Flash chromatography (SiO)₂2.5X 20cm, 2-5% methanol-chloroform gradient) followed by HPLC (YMC S-10, ODS, 30X 500mm column eluting with methanol at 20 ml/min) gave the desired cis-acetal product, compound I (0.60g, 43%) and the undesired diastereomer (0.32g, 22%) as well as starting compound E (. about.10%).

MS(ESI⁺)：879.3(2M+Na)⁺，451.2(M+Na)⁺，429.2(M+H)⁺；MS(ESI)：427.3(M-H)^-.

By the subsequent ester derivative (for the synthesis of epothilone C)¹³C and¹the stereochemistry of H NMR can be demonstrated by direct comparison with the same intermediates described in k.c. nicolaou et al, angelw.chem.int.ed.engl. (applied chemistry, international english edition), 1997, 36, 166.

(S) -2- [ N- [ tert-butoxy) carbonyl ] amino ] -4-pentenoic acid

The reaction mixture was washed successively with sodium bicarbonate (6.6g, 78mmol) and di-tert-butyl dicarbonate (10.4g,1.8mmol) L-2-amino-4-pentenoic acid (NovaBiochem, 3.0g, 26mmol) in THF-H₂O (1: 1, 200mL) solution. The reaction mixture was warmed to 25 ℃ and stirred for 16 hours. The pH of the mixture was adjusted to 4 by careful addition of saturated aqueous citric acid solution at 0 ℃ and the mixture was extracted with ethyl acetate (4X 50 mL). The combined organic layers were washed with saturated aqueous sodium chloride (75mL), dried (sodium sulfate) and concentrated in vacuo. By flash chromatography (SiO)₂4.0X 6cm, 5-10% methanol-chloroform gradient elution) to give Compound J (5.5g, 99%) as a colorless oil. MS (ESI)^-)：429.3(2M-H)^-，214.1(M-H)^-。

K.(S)-2-[N²- [ (tert-butoxy) carbonyl]Amino group]-N-methoxy-N-methyl-4-pentenamide

A solution of compound J (2.9g, 13mmol) in chloroform (55mL) was treated with N, O-dimethylhydroxylamine hydrochloride (1.4g, 15mmol), 1-hydroxybenzotriazole (2.0g, 15mmol), 4-methylmorpholine (4.4mL, 40mmol), and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (3.4g, 18mmol) in that order at 0 ℃. The reaction mixture was gradually warmed to 25 ℃, stirred for 16 hours, and diluted with water (100 mL). The phases were separated and the aqueous phase was extracted with ethyl acetate (3X 75 mL). The combined organic phases were washed with 5% aqueous hydrochloric acid (100mL), saturated aqueous sodium bicarbonate (100mL), saturated aqueous sodium chloride (100mL), dried (sodium sulfate), and concentrated in vacuo. By flash chromatography (SiO)₂3.0X 20cm, gradient elution 25-50% ethyl acetate-hexanes) to give compound K (2.5g, 71%) as a colorless oil. MS (ESI)⁺)：258.9(M+H)⁺202.9 (M-isobutylene), 158.9 (M-BOC); MS (ESI)^-)：257.2(M-H)^-。

L. (S) -3- [ N- [ (tert-butoxy) carbonyl ] amino ] -5-hexen-2-one

A solution of compound K (2.5g, 1.0mmol) in THF (65mL) was treated with methylmagnesium bromide (3.0M in ether, 8.1mL, 2.4mmol) at 0 deg.C. The reaction mixture was stirred at 0 deg.C (2.5 hours) and carefully poured into a saturated aqueous solution of ammonium chloride (100 mL). The two phases are separated and extracted with ethyl acetate (3 recovery)75mL) of the aqueous phase. The combined organic extracts were washed with saturated aqueous ammonium chloride (75mL), water (75mL), saturated aqueous sodium chloride (75mL), dried (magnesium sulfate), and concentrated in vacuo. By flash chromatography (SiO)₂3.0X 20cm, 10-25% ethyl acetate-hexanes gradient) to give (S) -2- [ N- [ (tert-butoxy) carbonyl ] as a colorless oil]Amino group]-5-hexen-2-one (2.2g, 67%). MS (ESI)⁺)：213.9(M+H)⁺157.9 (M-isobutene), 113.9 (M-BOC); MS (ESI)^-)：212.2(M-H)^-。

(S) -4- [3- [ N- [ (tert-butoxy) carbonyl ] amino ] -2-methyl-1 (E), 5-hexadienyl ] -2-methylthiazole

A solution of 2-methyl-4-thiazolylmethyl diphenylphosphine oxide (2.5g, 8.0mmol, Danishefsky et al J.Org.Chem. (J.org. chem., 1996, 61, 7998) in THF (38mL) is treated dropwise over 5 minutes at-78 ℃ with n-butyllithium (1.6M in hexane, 5.2mL, 8.4 mmol). The resulting bright orange mixture was stirred at-78 ℃ for 15 minutes and treated with compound L (0.81g, 3.8mmol) in THF (5 mL). After standing at-78 ℃ for 10 minutes, the cooling bath was removed and the reaction mixture was warmed to 25 ℃ (2 hours). The mixture was poured into a saturated aqueous solution of ammonium chloride (50mL), and the two phases were separated. With Et₂The aqueous phase was O (3X 50mL) extracted and the combined organic extracts were washed successively with water (75mL), saturated aqueous sodium bicarbonate (75mL), saturated aqueous sodium chloride (75mL), dried (sodium sulfate), and concentrated in vacuo. By flash chromatography (SiO)₂4.0X 30cm, 10-20% ethyl acetate-hexanes gradient) to give compound M (0.23g, 18%) as a colorless oil along with the recovered starting ketone (20-30%). MS (ESI)⁺)：309.1(M+H)⁺253.0 (M-isobutene); MS (ESI)^-)：307.3(M-H)^-。

(S) -4- (3-amino-2-methyl-1 (E), 5-hexadienyl) -2-methylthiazole

Compound M (0.15g, 0.49mmol) was treated with 4.0N 1, 4-dioxane hydrochloride solution (5mL) at 0 ℃ under argon (30 min). The volatiles were removed in vacuo and the resulting white foamy mass was dissolved in cold sodium bicarbonateSaturated aqueous solution (3 mL). The solution was extracted with ethyl acetate (4 × 10mL), the combined ethyl acetate layers were dried (sodium sulfate) and concentrated in vacuo. By flash chromatography (SiO)₂1.0X 5cm, 5-10% methanol-chloroform gradient elution) to give compound N (88mg, 88%) as a colorless oil. MS (ESI)⁺)：209.0(M+H)⁺；MS(ESI^-)：207.2(M-H)^-。

O. (3S, 6R, 7S, 8S) -N- (S) - [1- (2-methyl-4-thiazolyl) -2-methyl-1 (E), 5-hexadien-3-yl ] -3-tert-butyldimethylsilyloxy-4, 4, 6, 8-tetramethyl-7-hydroxy-5-oxo-12-tridecenamide

Compound M (88mg, 0.42mmol) in DMF (1.3ml) was treated with Compound I (0.15g, 0.35mmol), 1-hydroxybenzotriazole (49mg, 0.36mmol), and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (0.10g, 0.52mmol) in that order at 0 ℃. The reaction mixture was gradually warmed to 25 ℃, stirred for 15 hours, and diluted with water (3 mL). The mixture was extracted with ethyl acetate (3 × 10mL), and the combined organic phases were washed with 5% aqueous hydrochloric acid (10mL), saturated aqueous sodium bicarbonate (10mL), saturated aqueous sodium chloride (10mL), dried (sodium sulfate), and concentrated in vacuo. By flash chromatography (SiO)₂1.5X 20cm, 2.5% methanol-chloroform) to give compound O as a white foam (0.17g, 77%). MS (ESI)⁺)：619.3(M+H)⁺。

P.[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4-tert-butyldimethylsilyloxy-8-hydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13 (E) -cyclohexadecene-2, 6-dione

A solution of compound O (17mg, 27mmol) in degassed benzene (8.0mL) was treated with Grubb's catalyst [ bis (tricyclohexylphosphine) benzylideneruthenium dichloride (Strem Chemicals, 11mg, 14mmol) under argon. The reaction mixture was stirred at 25 ℃ for 15 h and treated with another portion of catalyst (5.0mg, 4.5 mmol). After a further 7 hours, the benzene was removed in vacuo and the black viscous residue passed through a pad of silica gel (1.0)X 3cm), eluting with diethyl ether (25 mL). The eluate was concentrated in vacuo to give a 5: 1(E/Z) mixture of separable geometric isomers. By PTLC (SiO)₂1mm plate, eluting twice with hexane-toluene-ethyl acetate 1: 1 solution) to give the E-isomer of Compound P (5.1mg, 34%) and the corresponding Z-isomer (1.0mg, 6.7%). For compound P: MS (ESI)⁺)：1181.7(2M+H)⁺，591.4(M+H)⁺. For the Z-isomer: MS (ESI)⁺)：1181.5(2M+H)⁺，613.2(M+Na)⁺，591.2(M+H)⁺；MS(ESI^-)：589.3(M-H)^-。

Q.[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13 (E) -cyclohexadecene-2, 6-dione

Trifluoroacetic acid (0.1mL) was added to a 1 dram vial containing a solution of compound P (2.3mg, 3.9mmol) in dichloromethane (0.4mL) at 0 ℃. The reaction mixture was sealed under Ar atmosphere and stirred at 0 ℃. After 4 hours, the volatiles were removed under a constant argon flow at 0 ℃. To the residue were added a saturated aqueous solution of sodium hydrogencarbonate (1mL) and ethyl acetate (1mL), and the two layers were separated. The aqueous phase was extracted with ethyl acetate (4 × 1mL), the combined ethyl acetate layers were dried (sodium sulfate) and concentrated in vacuo. By PTLC (SiO)₂20X 10X 0.025cm, 5% methanol-chloroform) to give [4S- [4R ] as a white thin film^*，7S^*，8S^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13 (E) -cyclohexadecene-2, 6-dione (1.3mg, 68%). MS (ESI)⁺)：953.5(2M+H)⁺，477.3(M+H)⁺；MS(ESI^-)：475.5(M-H)^-。

Example 2

The following compounds can be prepared according to the above reaction scheme:

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 13, 17-trioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 13, 17-trioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1, 10-dioxa-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1, 10-dioxa-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 14, 17-trioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 14, 17-trioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-bisHydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1, 11-dioxa-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1, 11-dioxa-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecan-9-one;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecan-9-one;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-3, 8, 8, 10, 12, 16-hexamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-3, 8, 8, 10, 12-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13, 16-hexamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-oxa-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 16-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-oxa-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-6, 8, 8, 10, 12, 16-hexamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-6, 8, 8, 10, 12-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-4, 8, 8, 10, 12, 16-hexamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-4, 8, 8, 10, 12-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-1, 5, 5, 7, 9, 13-hexamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-1, 5, 5, 7, 9-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-13-aza-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-13-aza-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-10-aza-1-oxa-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-10-aza-1-oxa-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-14-aza-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-14-aza-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-11-aza-1-oxa-13-cyclohexadecene-2, 6-dione;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-11-aza-1-oxa-13-cyclohexadecene-2, 6-dione;

[1S-[1R^*，3R^*，7R^*，10S^*，11R^*，12R^*，16S^*]]-N-phenyl-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-5, 9-dioxo-4, 17-dioxabicyclo [14.1.0 ]]Heptadecane-3-carboxamide;

[1S-[1R^*，3R^*，7R^*，10S^*，11R^*，12R^*，16S^*]]-N-phenyl-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-5, 9-dioxo-4, 17-dioxabicyclo [14.1.0]Heptadecane-3-carboxamide;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*]]-N-phenyl-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-2, 6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide;

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*]]-N-phenyl-4, 8-dihydroxy-5, 5, 7, 9-tetramethyl-2, 6-dioxo-1-oxa-13-cyclohexadecene-16-carboxamide;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) cyclopropyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione;

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) cyclopropyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione.

Example 3

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-azido-12, 13-epoxy-4, 4, 6, 8, 12, 16-hexamethyl-7-hydroxy-17- (2-methyl-4-thiazolyl) -5-oxo-16-heptadecenoic acid

A solution of epothilone B (0.35g, 0.69mmol) in degassed THF (4.5mL) was treated with a catalytic amount of tetrakis (triphenylphosphine) palladium (0) (80mg, 69mmol) and the suspension was stirred at 25 ℃ under Ar for 30 min. The resulting bright yellow homogeneous solution was immediately treated with a solution of sodium azide (54mg, 0.83mmol) in degassed water (2.2 mL). The reaction mixture was incubated at 45 ℃ for 1 hour, diluted with water (5mL) and extracted with ethyl acetate (4X 7 mL). The organic extract was washed with saturated aqueous sodium chloride (15mL) and dried (sulfuric acid)Sodium), concentrated in vacuo. By flash chromatography (SiO)₂3.0X 15cm, 95: 5.0: 0.5 chloroform-methanol-acetic acid) to give Compound A (0.23g, 61%) as a colorless oil. MS (ESI)⁺)：551(M+H)⁺；MS(ESI^-)：549(M-H)^-。

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-amino-12, 13-epoxy-4, 4, 6, 8, 12, 16-hexamethyl-7-hydroxy-17- (2-methyl-4-thiazolyl) -5-oxo-16-heptadecenoic acid

A solution of Compound A (0.23g, 0.42mmol) in THF (4.0mL) was treated with water (23mL, 1.25mmol) and triphenylphosphine on polymer (Aldrich, polystyrene cross-linked with 2% DVB, 0.28g, 0.84mmol) at 25 ℃. The suspension was stirred at 25 ℃ under Ar (32 h), filtered through a pad of Celite and concentrated in vacuo. By flash chromatography (SiO)₂1.5X 10cm, 95: 5.0: 0.5 to 90: 10: 1.0 chloroform-methanol-acetic acid gradient) to give Compound B (96mg, 44%) as a colorless oil. MS (ESI)⁺)：525.2(M+H)⁺；MS(ESI^-)：523.4(M-H)^-。

Alternatively, under Ar, under a mixture of Compound A (0.26g, 0.47mmol) and PtO₂(0.13g, 50 wt%) in a 25mL round-bottom flask was added absolute ethanol. Subjecting the resulting black mixture to one atmosphere of H₂Stirred for 10 hours and then with N₂Cleaning system, adding another part of PtO₂(65mg, 25 wt%). The reaction mixture is then brought to one atmosphere of H₂Stirred for 10 hours. Then using N₂The system was washed and the reaction mixture was filtered through a pad of Celite eluting with methylene chloride (3X 25 mL). The solvent was removed in vacuo and the residue was purified as above to give compound B (0.19g, 75%).

Alternatively, a solution of compound A (20mg, 36mmol) in THF (0.4mL) was treated with triphenylphosphine (19mg, 73mmol) under Ar. The reaction mixture was warmed to 45 ℃, stirred for 14 hours, and cooled to 25 ℃. The resulting iminophosphorane was treated with ammonium hydroxide (28%, 0.1mL) and the reaction mixture was warmed to 45 ℃. After 4 h, the volatiles were removed in vacuo and the residue was purified as above to give compound B (13mg, 70%).

C.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

A solution of compound B (0.33g, 0.63mmol) in degassed DMF (250mL) was treated with solid sodium bicarbonate (0.42g, 5.0mmol) and diphenylphosphorylazide (0.54mL, 2.5mmol) at 0 deg.C under Ar. The resulting suspension was stirred at 4 ℃ for 24 hours, diluted with phosphate buffer (250mL, pH 7) at 0 ℃ and extracted with ethyl acetate (5 × 100 mL). The organic extracts were washed with 10% aqueous lithium chloride (2X 125mL), dried (sodium sulfate) and concentrated in vacuo. The residue was first purified by flash chromatography (SiO)₂2.0X 10cm, 2-5% methanol-chloroform gradient elution) and then passed through a Chromatotron (2mm SiO. sub.2 mm)₂CF rotor, 2-5% methanol-chloroform gradient elution) to give the title compound as a colorless oil (0.13g, 40%).

¹H NMR(CDCl₃400MHz δ 6.98(s, 1H), 6.71(d, 1H, NH, J ═ 8.1Hz), 6.56(s, 1H), 4.69-4.62(m, 1H), 4.18-4.12(m, 1H), 4.01-3.96(m, 1H), 3.86(s, 1H), 3.38-3.34(m, 1H), 2.82(dd, 1H, J ═ 5.6, 6.0Hz), 2.71(s, 3H), 2.58(s, 1H), 2.43(dd, 1H, J ═ 9.0, 14.5Hz), 3.34(dd, 1H, J ═ 3.0, 14.5Hz), 2.14(s, 3H), 2.05-1.92(m, 2H), 1.82-1H (s, 1H), 1H, 7.41 (H), 7.6H, 6.0 (s, 3H), 3.00 (s, 3H), 3H, 8(s, 3H), 3H, 3.00 (d, 8, 3H, 8H, 3H, J ═ 8H, 6.00 (d, 3H); MS (ESI)⁺)：507.2(M+H)⁺；MS(ESI^-)：505.4(M·H)^-.

Example 4

Method for reducing epothilone and epothilone analogue oxirane ring

A two-necked flask was charged with magnesium turnings (24mg, 1.0 mmol). The flask was flame dried under vacuum and cooled under argon. Bis (cyclopentadienyl) titanium dichloride (250mg, 1.0mmol) was added first followed by anhydrous THF (5 mL). The suspension was drawn under low vacuum with stirring and the reaction flask was refilled with argon. The red suspension turned black and after 1.5 hours turned a uniform dark green color, with almost all of the magnesium metal being consumed. An aliquot (3.5mL, 0.70mmol, 3.5eq) was removed and cooled to-78 ℃ under argon. To this solution was added epothilone A (99mg, 0.20mmol, 1.0 eq). The reaction mixture was warmed to room temperature and stirred for 15 minutes. The volatiles were removed in vacuo and the residue was chromatographed twice on silica gel (25g) eluting with 35% ethyl acetate/hexanes to give 76mg (80%) of epothilone C as a pale yellow viscous oil.

Example 5

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-azido-3, 7-dihydroxy-12, 13-epoxy-4, 4, 6, 8, 16-pentamethyl-17- (2-methyl-4-thiazolyl) -5-oxo-16 (E) -heptadecenoic acid

Tetrakis (triphenylphosphine) palladium (0) (1.17g, 1.01mmol, 0.10eq) was added to a solution of epothilone A (4.97g, 10.1mmol, 1.0eq) in degassed THF (100mL) at room temperature and the mixture was stirred under argon for 30 min. To the above reaction mixture was added sodium azide (0.980g, 15.1mmol, 1.5eq) and degassed water (10mL) was added. The reaction mixture was incubated at 45 ℃ for 1 hour, cooled to room temperature, diluted with ethyl acetate (300mL) and then with water (150 mL). Mixing the water layerExtract with ethyl acetate (3X 100 mL). The combined organic extracts were washed with brine (150mL), dried (sodium sulfate), filtered, and concentrated in vacuo. The oily residue was purified by flash chromatography on silica gel (eluting with 0.1% acetic acid in 0-5% methanol/chloroform) to give compound a as a glass solid (1.84g, 34.0% yield). MS (ESI)⁺)：537(M+H)⁺；MS(ESI^-)：535(M-H)^-。

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-amino-3, 7-dihydroxy-12, 13-epoxy-4, 4, 6, 8, 16-pentamethyl-17- (2-methyl-4-thiazolyl) -5-oxo-16-heptadecenoic acid

Platinum oxide (0.980g, 4.30mmol, 1.25eq) was added to a solution of compound a (1.85g, 3.44mmol, 1.0eq) in absolute ethanol (137 mL). The reaction mixture was stirred vigorously in a hydrogen hood at room temperature for 16 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The oily residue was purified by preparative HPLC (YMC S-15ODS 50 × 500mm column 45 min/gradient, 0-100% B, 50 ml/min, retention time 17 min, a 0.1% acetic acid/5% acetonitrile/95% water, B0.1% acetic acid/5% water/95% acetonitrile). The appropriate fractions were concentrated in vacuo and the residue was lyophilized in aqueous acetonitrile to give compound B as a colourless solid (1.33g, 76.0% yield). MS (ESI)⁺)：511(M+H)⁺；MS(ESI^-)：509(M-H)^-。

C.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12-tetramethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

Compound B (0.860g, 1.68mmol, 1.0eq) was dissolved in anhydrous DMF (0.00250M, 672mL) and degassed at room temperature for one hour. The solution was cooled to 0 ℃ and solid sodium bicarbonate (1.13g, 13.4mmol, 4.0eq) and diphenylphosphoryl azide (1.85g, 6.72mmol, 8.0eq) were added under argon. The reaction mixture was stirred at 4 ℃ under argon for 16 hours. The reaction mixture is then cooledThe reaction was quenched by slow addition of pH 7 phosphate buffer (400mL) while cooling to-60 ℃. The temperature was kept below-30 ℃. The mixture was slowly warmed to room temperature and extracted with ethyl acetate (1 l). The aqueous layer was washed with ethyl acetate (4X 300 mL). The organic extracts were combined, washed with 10% aqueous lithium chloride (500mL), dried (sodium sulfate), filtered, and concentrated in vacuo. The oily residue was purified by preparative HPLC (YMC S-15ODS 50 × 500mm column 45 min/gradient, 0-100% B, 50 ml/min, retention time 35 min, a 5% acetonitrile/95% water, B5% water/95% acetonitrile). The appropriate fractions were concentrated in vacuo and the residue was lyophilized in aqueous acetonitrile to give the title compound as a colorless solid (0.220g, 26.0% yield). MS (ESI)⁺)：493(M+H)⁺；MS(ESI^-)：491(M-H)^-。

Example 6

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-1-aza-13 (Z) -cyclohexadecene-2, 6-dione;

tungsten hexachloride (0.19g, 0.49mmol, 0.5 equiv.) was dissolved in THF (5.0mL) and the solution was cooled to-78 ℃. N-butyllithium in hexane (1.6M, 0.63mL, 1.0mmol, 1.0 equiv) was added in one portion and the reaction mixture was warmed to room temperature over 20 minutes (the solution turned dark green when warmed to room temperature). A0.1M solution of the prepared tungsten reagent (0.79mL, 0.079mmol, 2.0 equiv.) was added to Compound 4C (0.020g, 0.039mmol, 1.0 equiv.) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes, then quenched with saturated sodium bicarbonate (2.0 mL). The quenched solution was diluted with water (10mL) and the solution was extracted with dichloromethane (4X 20 mL). The combined organic extracts were dried (sodium sulfate), filtered, and concentrated in vacuo. Make the residue pass throughThe mineral was removed by column chromatography on silica gel (elution with 19/1 chloroform/methanol). The eluate was concentrated in vacuo. The residue was purified by phplc (YMC S5 ODS 30-100% B, 5% aqueous acetonitrile a 95% aqueous acetonitrile B, 3 ml/min, 220nm, 30 min gradient elution) and the appropriate fractions were concentrated in vacuo. The viscous solid was lyophilized in aqueous acetonitrile to give the title compound as a white solid (4.3mg, 29%). TLC: rf ═ 0.57(9/1 chloroform/methanol, observed by UV); HRMS: (M + H)⁺The calculated value was 491.29436, and the measured value was 491.2934.

Example 7

[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

A.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-methyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione, N-oxide

A solution of epothilone B (2.0g, 3.9mmol) in dichloromethane (30mL) was treated with 3-chloroperoxybenzoic acid (1.0g, 5.9mmol) at 25 deg.C under Ar for 2 hours. An additional 0.5g (3.0mmol) of 3-chloroperoxybenzoic acid was added and the reaction mixture was stirred for 2 hours. The reaction mixture was filtered and the filtrate was concentrated in vacuo. The residue was dissolved in ethyl acetateThe ester (100mL) was washed with saturated aqueous sodium bicarbonate (75mL), 5% aqueous sodium sulfite (75mL), water (75mL), dried (sodium sulfate), and concentrated in vacuo. By flash chromatography (SIO)₂4.5X 30cm, 2-10% methanol-chloroform gradient elution) to give compound A as a white solid (1.04g, 50%). MS (ESI)⁺)：524.3(M+H)⁺；MS(ESI^-)：522.5(M-H)^-。

B.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl]-4, 17-dioxabicyclo [14.1.0]Heptadecane-5, 9-dione, [ epothilone F]

2, 6-lutidine (0.82mL, 7.0mmol) and trifluoroacetic anhydride (0.87mL, 6.2mmol) were added under Ar to a resealable tube containing a solution of Compound A (0.46g, 0.88mmol) in dichloromethane (10 mL). The reaction vessel was sealed under Ar, heated to 75 ℃ (12 minutes), cooled to 25 ℃, and the volatiles were removed under a constant stream of nitrogen. The reaction tube was placed in a high vacuum pump for 15 minutes. The resulting residue was dissolved in methanol (10mL) and washed with ammonium hydroxide (28-30% NH)₄Water, 1.0 mL). The mixture was heated to 45 deg.C (10 minutes) and the volatiles were removed in vacuo. The crude reaction mixture was purified by HPLC (YMC S-15ODS 30 × 500mm column, 50% acetonitrile-water gradient free elution, flow rate 20 ml/min, retention time 28 min). The appropriate fractions were concentrated in vacuo and the residue was lyophilized in aqueous acetonitrile to give the title compound B as a white solid (0.22g, 48%). MS (ESI)⁺)：524.3(M+H)⁺，1047.6(2M+H)⁺；MS(ESI^-)：522.5(M-H)^-。

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-azido-3, 7-dihydroxy-12, 13-epoxy-4, 4, 6, 8, 12, 16-hexamethyl-17- (2-hydroxymethyl-4-thiazolyl) -5-oxo-16 (E) -heptadecenoic acid

With catalytic amount of tetrakis (triphenylphosphine) palladium (0) (40mg, 3.4X 10)^-2mmol) Compound B (0.18g, 0.34mmol) in degassed THF (3.0mL) and the suspension stirred at 25 ℃ under argon for 30 min. The resulting pale yellow homogeneous solution was treated by the addition of a solution of sodium azide (27mg, 0.41mmol) in degassed water (1.5mL) in one portion. The reaction mixture was incubated at 45 ℃ for 1 hour, diluted with water (5mL) and extracted with ethyl acetate (4X 10 mL). The organic extracts were washed with saturated aqueous sodium chloride (15mL), dried (sodium sulfate) and concentrated in vacuo. By silica gel flash chromatography (SiO)₂2.5X 15cm, 95: 5 chloroform-methanol to 95: 5.0: 0.5 chloroform-methanol-acetic acid gradient) to give Compound C as a colorless oil (39mg, 20%). MS (ESI)⁺)：567.4(M+H)⁺，1133.6(2M+H)⁺；MS(ESI^-)：565.5(M-H)^-，1131.8(2M-H)^-。

(3S, 6R, 7S, 8S, 12R, 13S, 15S) -15-amino-3, 7-dihydroxy-12, 13-epoxy-4, 4, 6, 8, 12, 16-hexamethyl-17- (2-hydroxymethyl-4-thiazolyl) -5-oxo-16 (E) -heptadecenoic acid

Under Ar, in the presence of compound C (40mg, 71mmol) and PtO₂(12mg, 30 wt%) to a 10mL round-bottom flask was added absolute ethanol (3 mL). The resulting black mixture was stirred under one atmosphere of hydrogen for 10 hours. The system was then purged with nitrogen and the reaction mixture was filtered through a nylon membrane (washing with 25mL of methanol). VacuumThe solvent was removed to give foam compound D (29mg, 76%). It was pure enough for the next reaction. LCMS: 541.3(M + H)⁺。

E.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

A solution of compound D (29mg, 54mmol) in degassed DMF (21mL) was treated with solid sodium bicarbonate (36mg, 0.43mmol) and diphenylphosphorylazide (46mL, 0.21mmol) at 0 deg.C under argon. The resulting suspension was stirred at 4 ℃ for 19 h, cooled to-40 ℃, diluted with 25mL of pH 7 phosphate buffer (added slowly, keeping the internal temperature below-30 ℃), extracted with ethyl acetate (4 × 10mL), and the organic extracts washed with cold 10% aqueous lithium chloride (25mL), dried (sodium sulfate), and concentrated in vacuo. By Chromatotron (1mm SiO)₂GF rotor, 2-5% methanol-chloroform gradient elution) to give the title compound E (9.1mg, 34%) as a colorless oil. MS (ESI)⁺)：523.2(M+H)⁺；MS(ESI^-)：521.5(M-H)^-。

Example 8

[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl]-1-aza-13 (Z) -cyclohexadecene-2, 6-dione

A.[1S-[1R^*，3R^*(E)，7R^*，10S^*，11R^*，12R^*，16S^*]]-7, 11-dihydroxy-8, 8, 10, 12, 16-pentamethyl-3- [ 1-methyl-2- (2-tert-butyldiphenylsiloxymethyl-4-thiazolyl) ethenyl]-4-aza-17-oxabicyclo [14.1.0]Heptadecane-5, 9-dione

A solution of compound 7E (6.8mg, 13mmol) in dichloromethane (0.5mL) was treated with triethylamine (2.7mL, 20mmol), 4-N, N-dimethylaminopyridine (0.2mg, 1.3mmol) and tert-butyldiphenylchlorosilane (3.7mL, 14mmol) at 0 deg.C under argon. The reaction mixture was gradually warmed to 25 deg.C (1 hour), cooled to 0 deg.C, quenched by the addition of saturated aqueous sodium bicarbonate solution (1mL), and extracted with ethyl acetate (4X 2 mL). The combined organic extracts were washed with brine (5mL), dried (sodium sulfate) and concentrated in vacuo. By flash chromatography (SiO)₂1.0X 5cm, 2-5% methanol-chloroform gradient elution) to give compound A (7.0mg, 71%) as a colorless oil. MS (ESI)⁺)：761.5(M+H)⁺；MS(ESI^-)：759.7(M-H)^-。

B.[4S-[4R^*，7S^*，8R^*，9R^*，15R^*(E)]]-4, 8-dihydroxy-5, 5, 7, 9, 13-pentamethyl-16- [ 1-methyl-2- (2-hydroxymethyl-4-thiazolyl) ethenyl]-1-aza-13 (Z) -cyclohexadecene-2, 6-dione

A solution of tungsten (IV) chloride (0.10g, 0.25mmol) in dry THF was treated with n-butyllithium (1.6M in hexane, 0.32mL, 0.50mmol) at-78 deg.C under Ar. Within 40 minutes willThe reaction mixture was warmed to 25 ℃ and then cooled to 0 ℃. An aliquot (0.2mL, 20mmol) of the resulting dark green homogeneous solution was added to a 1 dram vial containing Compound A (7.0mg, 9.2mmol) at 0 deg.C under Ar. The reaction mixture was warmed to 25 ℃, stirred for 30 minutes, quenched by the addition of saturated aqueous sodium bicarbonate (0.5mL), and extracted with ethyl acetate (4 × 1 mL). The combined organic extracts were dried (sodium sulfate) and concentrated in vacuo. By preparative TLC (SiO)₂20X 0.025cm, eluting with 5% methanol-chloroform) to give an unseparated mixture of the silyl-protected B (13Z) isomer of compound and a small (< 10%) minor (13E) isomer, which is directly subjected to the next deprotection step.

The silyl-protected isomer mixture of Compound B (2.3mg, 3.1mmol) was treated with 0.3ml of HF-pyridine in THF (2: 1: 0.5 THF/pyridine/HF-pyridine solution, Aldrich Chemical Co.) at 25 deg.C. After 1 hour, the reaction mixture was neutralized with a saturated aqueous solution of sodium hydrogencarbonate (0.5mL), and extracted with ethyl acetate (4X 1 mL). The combined organic extracts were washed with a saturated aqueous solution of sodium bicarbonate (1mL), dried (sodium sulfate) and the volatiles removed in vacuo. By preparative TLC (SiO)₂20 × 10 × 0.025cm, eluting with 5% methanol-chloroform) to give the title compound (13Z isomer) as a thin film, and a mixture of a small amount of the unseparated (< 10%) minor (13E) isomer (0.96mg, 20%, vs. two steps). MS (ESI)⁺)：507.3(M+H)⁺；MS(ESI^-)：505.6(M-H)^-。

Claims (9)

1. A compound of the formula

Wherein Q is selected from

G is

W is NH;

x is O or H, H;

y is O;

B₁and B₂Is OH;

R₁，R₂，R₃，R₄，R₅，R₇and R₁₂Selected from H and alkyl;

R₆is methyl;

R₁₁is that

Or

Wherein R is H, lower alkyl, or lower alkyl substituted with OH;

R₈is hydrogen or methyl;

and is

Wherein the "alkyl" refers to a straight or branched unsubstituted hydrocarbon group having 1 to 20 carbon atoms;

wherein said "lower alkyl" refers to an alkyl group having 1 to 4 carbon atoms.

2. The compound of claim 1, wherein

Q is

And R₈Is methyl; r is methyl or hydroxymethyl.

3. Use of a compound of any one of claims 1-2 for the manufacture of a medicament for treating a disease in a patient; wherein the disease is selected from the group consisting of:

-cancers of the bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; squamous cell carcinoma;

-leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphomas, T-cell lymphomas, hodgkin lymphomas, non-hodgkin lymphomas, hairy cell lymphomas and burkitt lymphomas;

-acute and chronic myeloid leukemia and promyelocytic leukemia;

fibrosarcoma and rhabdomyosarcoma;

-melanoma, seminoma, teratoma, neuroblastoma and glioma;

astrocytomas and schwannomas;

fibrosarcoma, rhabdomyosarcoma and osteosarcoma; and

-xeroderma pigmentosum, keratoacanthoma, thyroid follicular cancer;

follicular lymphoma, carcinomas associated with mutations in the p53 gene, hormone-associated breast, prostate and ovarian tumors.

4. Use of a compound of any one of claims 1-2 for the manufacture of a medicament for treating cancer in a patient; wherein the cancer is selected from:

-cancers of the bladder, breast, colon, lung, ovary and pancreas;

-non-hodgkin's lymphomas;

-melanoma; and

-prostate tumors.

5. A compound having the formula:

wherein Q is selected from the group

And

R⁷is hydrogen or methyl; and

r is selected from hydrogen, alkyl or hydroxyalkyl, wherein the "alkyl" is as defined in claim 1.

6. The compound of claim 5 wherein Q is

R⁷Is methyl and R is methyl or hydroxymethyl.

7. A compound having the formula:

8. use of a compound of claim 7 for the manufacture of a medicament for treating a disease in a patient; wherein the disease is selected from the following:

-cancers of the bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, cervix, thyroid and skin; squamous cell carcinoma;

-leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia, B-cell lymphomas, T-cell lymphomas, hodgkin lymphomas, non-hodgkin lymphomas, hairy cell lymphomas and burkitt lymphomas;

-acute and chronic myeloid leukemia and promyelocytic leukemia;

fibrosarcoma and rhabdomyosarcoma;

-melanoma, seminoma, teratoma, neuroblastoma and glioma;

astrocytomas and schwannomas;

fibrosarcoma, rhabdomyosarcoma and osteosarcoma; and

-xeroderma pigmentosum, keratoacanthoma, thyroid follicular cancer;

follicular lymphoma, carcinomas associated with mutations in the p53 gene, hormone-associated breast, prostate and ovarian tumors.

9. Use of a compound of claim 7 for the manufacture of a medicament for treating cancer in a patient; wherein the cancer is selected from:

-cancers of the bladder, breast, colon, lung, ovary and pancreas;

-non-hodgkin's lymphomas;

-melanoma; and

-prostate tumors.