Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D225/00—Heterocyclic compounds containing rings of more than seven members having one nitrogen atom as the only ring hetero atom
  • C07D225/02—Heterocyclic compounds containing rings of more than seven members having one nitrogen atom as the only ring hetero atom not condensed with other rings
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D313/00—Heterocyclic compounds containing rings of more than six members having one oxygen atom as the only ring hetero atom
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
  • C07F7/02—Silicon compounds
  • C07F7/08—Compounds having one or more C—Si linkages
  • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
  • C07F7/1804—Compounds having Si-O-C linkages
• • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
  • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
  • Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
  • Y02P20/00—Technologies relating to chemical industry
  • Y02P20/50—Improvements relating to the production of bulk chemicals
  • Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Definitions

• the present invention relates to analogs of dictyostatin, intermediates for the synthesis of such analogs and methods of synthesis of such intermediates and analogs.
• References set forth herein may facilitate understanding of the present invention or the background of the present invention. Inclusion of a reference herein, however, is not intended to and does not constitute an admission that the reference is available as prior art with respect to the present invention.
• the discovery and development of new chemotherapeutic agents for the treatment of cancer is currently of high importance. Some of the best currently available chemotherapeutic agents are natural products or natural product analogs. For example, Taxol (paclitaxel) is a natural product that is currently being used to treat patients with breast and ovarian cancer among others.
• Taxol A number of analogs of Taxol, including Taxotere (docetaxel), are also powerful anticancer agents. [0006] Recently, the natural product (+)-discodermolide and its analogs have shown great promise as anticancer agents. Discodermolide has been shown to have a mechanism of action similar to Taxol, but it is active against Taxol-resistant cell lines and it is more water soluble than Taxol. Accordingly, it may have a different and/or broader spectrum of action than Taxol and be easier to formulate and administer. Analogs of discodermolide have been made and tested for activity. For example, see Myles, D. C. Emerging microtubule stabilizing agents for cancer chemotherapy, Annual Reports In Medicinal Chem; Academic Press: San Diego, CA, 2002; pp 125-132. An interesting feature of discodermolide is that both enantiomers are biologically active.
• R 1 is H, an alkyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom
• R 2 is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R a , R and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R 1 SiR a R b R c or a benzyl group, wherein R 1 is an alkylene group
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or , -NR s R h ,
• R ,23a a is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23b is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR R b R c , CH 2 OR d , or COR e , or R 23a and R 23 together form a portion of six- membered acetal ring incorporating CR l R u ;
• R 4 and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group; and
• R 5 is H or OR 2b , wherein R 2 is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ; provided that the compound is not dictyostatin 1.
• R ,23a , ⁇ R23b are H.
• R is OH or OSiR a- Rnb Rnc .
• C2-C3 E-sterioisomers of the compounds or their enantiomers are provided. [0014]
• one such intermediate is a compound of the following structure or its enantiomer.
• R is an alkyl group, an aryl group, an alkoxylalkyl group, -R ic Si ⁇ R»ar R>b Rr>c or a benzyl group wherein R 1 is an alkylene group; R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 23a is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23 is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23 and R 23b together form a portion of six- membered acetal ring incorporating CP R";
• R l and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group;
• R 5 is H or OR 2 , wherein R 2b is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ; and
• R 10 is H or alkyl.
• the compound has the following stereostructure, or its enantiomer:
• R 2d is H, C(O)CH 3 or C(O)NH 2 .
• R 1 is H, an alkyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 and R 2d are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 5 is H or OR 2b , wherein R 2b is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e ;
• R* and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group
• R 2c is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c ,
• the compound has the following stereostructure or its enantiomer:
• R 1 is alkenyl
• R 2 and R 2d are independently, H, OC(O)CH 3 or OC(O)NR g R h wherein R g and R h are independently H, an alkyl group or an aryl group
• R lla and R l l are H or together form a portion of a six-membered acetal ring containing C(H)(p-C 6 H 4 OCH 3 ) or C(CH 3 ) 2
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R 1 SiR a R R c or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R is H or OR , wherein R is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R lla and R llb are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , COR e , or R lla and R ll together form a portion of six-membered acetal ring containing CR l R u ;
• R l and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group;
• R 2c is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR , or COR e
• R 10 is H or alkyl
• R I4a and R I4b are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , COR e , or R 14a and R 14 together form a six-membered ring containing CR V R W , wherein R v and R are independently H, an alkyl group, an aryl group or an alkoxyaryl group.
• the compound has the following stereostructure or its enantiomer:
• R 10 is H or alkyl.
• R is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c ,
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R ll and R 11 are independently H, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c ,
• R 2c is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e , and R 10 is H or alkyl;
• R 14a and R 14 are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , COR e , or R 14 and R 14b together form a six-membered ring containing CR V R W , wherein R v and R are independently H, an alkyl group, an aryl group or an alkoxyaryl group.
• the compound has the following stereostructure or its enantiomer:
• R is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R a , R and R c are independently an alkyl group or an aryl group;
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R 2c is H, aprotecting group, an allcyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e , and R 10 is H or alkyl;
• R 16 is H or alkyl
• R is CH 2 OR , CHO, CO 2 R , wherein R is H, a protecting group, an allcyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ; and
• the compound has the following stereostructure or its enantiomer:
• a process for synthesizing dictyostatin analogs includes a process for conversion of a first compound having the following formula or its enantiomer:
• R 1 is H, an alkyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 is H, a protecting group, an allcyl group, a benzyl group, a trityl group, -SiR a R b R c ,
• R 2d is H
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR'Tl R c or a benzyl group, wherein R 1 is an alkylene group;
• R e is an allcyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R and R h are independently H, an alkyl group or an aryl group;
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 23 is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23 is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23a and R 23 together form a portion of six- membered acetal ring incorporating CR ⁇ R";
• R* and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group; and R 10 is H;
• the first compound has the following stereostructure or its enantiomer:
• R 1 is H, an alkyl group, an allcenyl group, an alkynyl group, or a halogen atom
• R 2 is H, an allcyl group, a benzyl group, a trityl group, -SiR R b R c , CH 2 OR d , or COR e
• R 2d is H
• R a , R and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R 1 SiR R b R c or a benzyl group, wherein R 1 is an alkylene group;
• R 1 is allcenyl
• the first compound is reacted with 2,4,6-trichlorobenzoylchloride.
• R a , R and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, ,a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• R lla and R ll are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , COR e , or R lla and R ll together form a portion of six-membered acetal ring containing CR l R u ;
• R l and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group;
• the present invention provides a compound having the following formula, or its enantiomer
• R is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a tntyl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R a , R b and R c are independently an allcyl group or an aryl group
• R d is an allcyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R g and R 1 are independently H, an allcyl group or an aryl group;
• R 11 is a protecting group, an allcyl group, and aryl group, a benzyl group, a trityl group,
• R 2c is H, a protecting group, an allcyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e , and R 10 is H or allcyl;
• R 16 is H or allcyl
• R is CH 2 OR , CHO, CO 2 R , wherein R is H, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e ; and
• the compound has the following stereostructure, or its enantiomer
• R 2 is H, an allcyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR , or COR e ;
• the present invention provides a compound having the following formula, or its enantiomer x r O OR 1 ⁇ 1 CHR12 '
• X is H, NCH 3 (OCH 3 ), or a leaving group
• R 11 is H, a protecting group, an alkyl group, and aryl group, a benzyl group, a trityl group,
• R l and R u are independently H, an allcyl group or an aryl group
• R 2c is H, a protecting group, an allcyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e , and R 10 is H or allcyl.
• R 10 is H or allcyl.
• R a and R llb are independently H, a protecting group, an alkyl group, and aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , COR e , or R lla and R ll together form a portion of six-membered acetal ring containing CR*R U ;
• R t and R u are independently H, an allcyl group, an aryl group or an alkoxyaryl group;
• R ,2 a protecting group, an allcyl group, an aryl group, a benzyl group, a trityl group,
• R , R and R c are independently an allcyl group or an aryl group
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• R 11 is an alkyl group, and aryl group, a benzyl group, a trityl group, -SiR a R R c , CH OR ,
• R 10 is H or alkyl
• R is CH OR , CO R , wherein R is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ; and R 24 is C ⁇ C to a compound of the following formula, or its enantiomer
• R l la is H, an allcyl group, and aryl group, a benzyl group, a trityl group, -SiR a R R c ,
• CH 2 OR d , COR e and R l l is an alkyl group, and aryl group, a benzyl group, a trityl group,
• R l and R u are independently H, an alkyl group or an aryl group
• the process includes at least the steps of semi-reduction of the alkyne, asymmetric reduction of the ketone and protection of a resulting alcohol, or asymmetric reduction of the ketone, protection of a resulting alcohol and semihydrogentation of the alkyne, or asymmetric reduction of the ketone, semi-hydrogentation of the alkyne and protection of a resulting alcohol.
• the present invention provides a compound of the following formula, or its enantiomer
• R >2 is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e ;
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R 1 SiR a R b R c or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• R a , R and R c are independently an alkyl group or an aryl group
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R R c or a benzyl group, wherein R 1 is an alkylene group;
• R g and R h are independently H, an alkyl group or an aryl group;
• R 16 is H or alkyl
• X is CI, Br or I with a second compound of the following formula, or its enantiomer
• X is NCH 3 (OCH 3 ), or a leaving group
• R 11 an alkyl group, and aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , COR e ;
• the present invention provides a process for reacting a first compound of the following formula, or its enantiomer
• R 2 a protecting group, an allcyl group, an aryl group, a benzyl group, a trityl group,
• R d is an allcyl group, an aryl group, an alkoxylalkyl group, -R ⁇ iR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an allcyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• R 16 is H or alkyl
• R 17 is CH 2 OR 2f , CHO, CO 2 R 10 , wherein R 2f is H, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R 11 an alkyl group, and aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , COR e ;
• R 1 is H, an alkyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 is H, a protecting group, an allcyl group, a benzyl group, a trityl group, -SiR a R R c ,
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 4 is (CH 2 ) P where p is an integer in the range of 4 to 12,
• R 5 is H or OR 2 , wherein R 2 is H, a protecting group, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR R R c , CH 2 OR d , or COR e ; and
• R 26 is H, a protecting group, an allcyl group, an aryl group, -SiR a R R c , or COR e ; [0038] In one embodiment, the compound of has the following stereostructure, or its enantiomer
• R ,23a is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23b is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e , or R 23a and R 23b together form a portion of six- membered acetal ring incorporating CR ⁇ R";
• R* and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group.
• R is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a ⁇ R)t> ⁇ R>c ,
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an allcyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R g and R h are independently H, an alkyl group or an aryl group;
• R 2 is H, a protecting group, an allcyl group, a benzyl group, a trityl group, -SiR a R R c ,
• R is an alkyl group, an aryl group, an alkoxylalkyl group, -R 1 SiR a R R c or a benzyl group, wherein R 1 is an alkylene group;
• R 3 is (CH 2 ) n where n is and integer in the range of 0 to 5, -CH 2 CH(CH 3 )-, -CH-CH-,
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 5 is H or OR 2 , wherein R 2b is H, an allcyl group, an aryl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e ; and
• R 26 is H, a protecting group, an alkyl group, a aryl group, -SiR a R R c , or COR e ; including the step of reacting a starting compound having the formula:
• the starting compound has the following structure, or its enantiomer
• R 1 is allcenyl;
• R 4
• R 3a is H, a protecting group, an allcyl group, a benzyl group, a trityl group, -SiR a R b R c , CH OR d , or COR e
• R 23 is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e , or R 23a and R 23 together form a portion of six- membered acetal ring incorporating CP R";
• R 4 and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group and the product compound has the following structure, or its enantiomer
• the present invention provides a process for converting a starting compound of the following structure
• R 1 is H, an allcyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 and R 2d are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R a , R b and R c are independently an alkyl group or an aryl group
• R d is an allcyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• R 4 is (CH ) P where p is an integer in the range of 4 to 12,
• the starting compound has the following structure, or its enantiomer
• R 1 is alkenyl
• R 4 is
• R 23a is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH OR d , or COR e
• R 23b is H, a protecting group, an allcyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e
• R 23a and R 23b together form a portion of six- membered acetal ring incorporating CR'R U ;
• R l and R u are independently H, an alkyl group, an aryl group or an alkoxyaryl group and the product compound has the following structure, or its enantiomer
• the present invention provides a compound of the following structure or its enantiomer
• R 1 is H, a protecting group, an allcyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 and R 2d are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R e is an alkyl group, an allyl group, a benzyl group, an aryl group, an alkoxy group, or
• R g and R h are independently H, an alkyl group or an aryl group;
• -CH C(CH 3 )-, or -C ⁇ C-;
• R ,23a is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e
• R 23b is H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e , or R 23a and R 23b together form a portion of six- membered acetal ring incorporating CR l R u ;
• R 10 is H or alkyl
• R is H, a protecting group, an allcyl group, an aryl group, an alkenyl group, an alkynyl group, or a halogen atom;
• R 2 and R 2d are independently H, a protecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , or COR e ;
• R a , R b and R c are independently an allcyl group or an aryl group;
• R d is an alkyl group, an aryl group, an alkoxylalkyl group, -R'SiR ⁇ R 0 or a benzyl group, wherein R 1 is an alkylene group;
• R g and R h are independently H, an alkyl group or an aryl group;
• -CH C(CH 3 )-, or -C ⁇ C-;
• R 5 is H or OR 2b , wherein R 2b is H, an alkyl group, an aryl group, a benzyl group, a trityl group, -SiR a R b R c , CH 2 OR d , or COR e ;
• R la and R 11 are independently H, a portecting group, an alkyl group, a benzyl group, a trityl group, -SiR a R R c , CH 2 OR d , COR e , or R lla and R ub together form a portion of six-membered acetal ring incorporating CR*R U ;
• R 4 and R u are independently H, an allcyl group, an aryl group or an alkoxyaryl group;
• R 2c is H, an allcyl group, a benzyl group, a trityl group, -SiR R b R c , CH 2 OR d , or COR e , and
• R 10 is H or alkyl
• alkyl group refers to a substituent on another group (for example, an alkyl group as a substituent of an alkylamino group or a dialkylamino group).
• aryl refers to phenyl or naphthyl.
• halogen or halo refer to fluoro, chloro, bromo and iodo.
• alkoxy refers to -OR, wherein R is an alkyl group.
• trityl refers to a triphenyl methyl group or -C(Ph) 3 .
• Certain groups such as amino and hydroxy groups may include protective groups as known in the art. Preferred protective groups for amino groups include tert- butyloxycarbonyl, formyl, acetyl, benzyl, /j-methoxybenzyloxycarbonyl, trityl.
• the present invention includes the synthesis of the compounds of the present invention as well as the biological assaying of such compounds and the biological activity of such compounds against, for example, cancer (such as breast, prostate cancer and ovarian cancer).
• cancer such as breast, prostate cancer and ovarian cancer
• the present invention provides a method of treating a patient for cancer, including the step of administering a pharmaceutically effective amount of a biologically active compound of the present invention or a pharmaceutically acceptable salt thereof.
• Figure 1 illustrates two embodiments of the synthesis of dictyostatin bottom fragment 15.
• Figure 2 illustrates two embodiments of the syntheses of dictyostatin middle fragment 29.
• Figure 3 illustrates one embodiment of the coupling of bottom and middle fragments of dictyostatin and elaboration to build the upper fragment.
• Figure 4 illustrates one embodiment of the construction of dictyostatin 1 and representative analogs 50 and 59.
• Figure 5 illustrates an embodiment of the synthesis of representative analog C 16-desmethyldictyostatin 79.
• Figure 6 illustrates an embodiment of the synthesis representative C6-epi,C14- epi intermediate 95.
• Figure 7 illustrates an embodiment of the synthesis of representative analog C2-E,C6-epi,C14-epi dictyostatin 100 and its C2-C3 Z-isomer.
• Figure 8 illustrates an embodiment of the synthesis of representative analog C6-epz ' ,C14-ep,C19-epi dictyostatin 108 and its C2-C3 E-isomer.
• Figure 9 illustrates representative examples and methods of synthesis of lactam analogs of the present invention.
• Figure 10 illustrates representative turbidity profiles of 16- desmethyldictyostatin in comparison to that of dictyostatin 1 in a tubulin-only (no MAPs, no GTP, assembly supported by monosodium glutamate) assay.
• Figures 1-9 show exemplary synthetic pathways and intermediates for the synthesis of dictyostatin analogs.
• the synthesis of an exemplary "bottom fragment" 15 for making dictyostatin and analogs is shown in Figure 1.
• 1,3-Propanediol 3 was elaborated via Evans chiral auxiliary-based methods to the known, bis-TBS-protected Homer-Wadsworth-Emmons product 10 in nine steps. See, Phukan, P.; Sasmal, S.; Maier, M. E. Eur. J. Org. Chem. 2003, 1733, and Andrus, M. B.; Argade, A. B. Tetrahedron Lett. 1996, 37, 5049.
• Alcohol 41 ⁇ was protected with a TBS group to give 42, whose PMP acetal was cleaved with DJJBAL-H to give alcohol 43 ( Figure 4).
• the allylic trityl group was removed with ZnBr 2 to give alcohol 45.
• Dess-Martin oxidation to the aldehyde and Still-Gennari reaction gave the (E,Z)-conjugated ester 46.
• the PMB group was removed with DDQ to give 47 and saponification with aqueous KOH in ⁇ tOH-THF to give acid 48.
• Analogs lacking the C9 oxygen atom can likewise be prepared by methods shown in that application. See, for example, Figures 8 and 11, among others.
• the preferred method for forming the macrolactones is the Yamaguchi lactonization. See, for example, Inanaga, J.; Kuniko, H.; Hirolco, S.; Katsuki, T.; Yamaguchi, M. Bull. Chem. Soc. Jpn. 1919, 52, 1989.
• An example of the Yamaguchi lactonization is the conversion of hydroxy acid 48 to lactone 49 in Figure 4.
• alkynyl ketone 32 For one example, see the conversion of alkynyl ketone 32 to alkynyl alcohol 33, to alkenyl alcohol 34, to silyl ether 35 in Figure 3.
• the preferred conditions for reduction of the ketone involve use of the Noyori reagent, see K. Matsumura, S. Hashiguchi, T. ariya, R. Noyori, J Am. Chem. Soc. 1997, 119, 8738-39.
• many other common ketone reducing agents both chiral and achiral, can also be used. See, for example, Itsuno, S. Enantioselective Reduction of Ketones. Org. React. (N.Y.) 1998, 52, 395-576.
• the preferred order is semi-reduction of the C10-C11 alkyne, followed by asymmetric reduction of the C9 ketone followed by (optionally) protection of the resulting alcohol.
• Other orders of reactions are asymmetric reduction of the ketone, protection of the alcohol and semi-reduction of the alkyne, or asymmetric reduction of the ketone, semi-reduction of the alkyne and protection of the alcohol.
• a preferred method is the deprotonation of the alkyne with a strong base, for example BuLi, followed by addition of a carboxylic acid derivative that is activated with a suitable leaving group.
• a strong base for example BuLi
• Preferred activated carboxylic acids for acylation are Weinreb amides where the leaving group is the N- methoxy-N-methyl amide group.
• Many other agents such as esters, acid halides, acid imidazolides, etc. can also be used. These have standard leaving groups such as alkoxide, imidazole and halide.
• the alkynyl anion can also be generated in situ from a silylalkyne by desilylation or from a geminal-haloalkene by treatment with two or more equivalents of a lithiating agent like BuLi.
• the alkynyl anion can be reacted with an aldehyde instead of an activated carboxylic acid to produce a C9 alcohol directly after workup. This route is more direct, but mixtures of epimers at C9 may result and chromatographic separation of the epimers may be required.
• One epimer of a C9 (or other) alcohol can be converted to the other by a Mitsunobu reaction.
• Lactam analogs of dictyostatin are important as anticancer agents because of their increased hydrolytic stability compared to the lactones, both in vivo and in vitro. These analogs are readily made by starting with intermediates of the current invention, as exemplified in Figure 9. Standard oxidation of the free C21 alcohol of 110 to a ketone followed by reductive amination provides 111. If desired, the C21 amine stereoisomers can be separated by chromatography. Hydrolysis of the ester to the acid followed by macrolactamization provides lactam 112.
• dictyostatin macrolactam 115 made for example by the sequence 113 -» 114 - 115, is exemplary of a lactam analog of this invention.
• the steps in the sequence can be conducted in different orders and also on different intermediates.
• the C21 nitrogen atom is installed earlier in the synthesis, it is optionally protected with a standard nitrogen protecting group for the subsequent steps prior to macrolactamization.
• this nitrogen can be installed by a Mitsunobu reaction of a suitably acidic nitrogen nucleophile (for example, azide) with a C21 alcohol. This reaction occurs with inversion, so the configuration of C23 is chosen accordingly.
• Tubulin polymerization The abilities of the new compounds to cause tubulin polymerization were determined under reaction conditions consisting of purified bovine brain tubulin (1 mg/mL) in the presence or absence of microtubule-associated proteins (MAPs, 0.75 mg/mL) and GTP (100 ⁇ M). Test agents were initially screened at 10 and 40 ⁇ M. In these experiments, test agent-induced assembly of soluble tubulin into polymer, with respect to the presence and absence of cofactors and at different temperatures, was monitored in a multi-cuvette, temperature-controlled spectrophotometer via development of turbidity in the solution.
• MAPs microtubule-associated proteins
• GTP 100 ⁇ M
• the initial temperature was closely controlled at 0 °C, then rapidly raised to 10 °C, to 20 °C, then finally to 30 °C to determine both the temperature at which a test agent induced assembly as well as the extent of agent-induced assembly.
• the temperature increases were followed by a rapid decrease in temperature back to 0 °C to determine the cold-stability of polymer formed.
• the effects of dictyostatin 1 and discodermolide 2 were similar and far more potent than those of paclitaxel.
• the C16-desmethyl compound 79 is especially potent among the analogs.
• Figure 10 shows the simplest of its turbidity profiles in comparison to that of dictyostatin 1 in a tubulin-only (no MAPs, no GTP, assembly supported by monosodium glutamate) assay wherein initial temperature was 0 °C for 2 min, followed by rapid rise in temperature to 30 °C for 20 min, then rapid decrease to 0 °C. Turbidity profiles showed that analogs 50 and 59 also caused tubulin assembly at temperatures lower than 30 °C. The results showed that all of the compounds had effects on the isolated target, tubulin, but with a range of potencies. [0091] Antiproliferative activity.
• C6- epz ' ,C14-epz ' -C19-ept-dictyostatin 108 and its C2E-diene derivative 109 were antiproliferative agents, giving mid micromolar GI50 values. Even though 100 also had three stereo/geometric alterations (C2E,C6-epi,C14-epi), it was a more potent antiproliferative agent than 108 and 109, showing high nanomolar GI50 values. With one notable exception (vide infra), the fold- resistance values for 1 and its analogs against 1A9/Ptxl0 and 1A9/Ptx22 cell lines were much lower than that observed for paclitaxel.
• test agents to inhibit the binding of radiolabeled forms of the microtubule stabilizers paclitaxel, discodermolide and epothilone B from tubulin polymer were determined.
• Dictyostatin 1 was equipotent to discodermolide in inhibition of the binding of radiolabeled paclitaxel and epothilone B to microtubules. These two compounds were the most potent of all agents tested.
• the open chain methyl ester 50 and the 16-desmethyl analog 79 were ca. 60% as potent as 1 in inl ibiting the binding of radiolabeled paclitaxel to microtubules.
• Test Agent (4 ⁇ M) [ 3 H]Paclitaxel [ 3 H]Discodermolide [ 14 C]Epothilone B dictyostatin (1) 75 ⁇ 5 (3) 40 + 3 (3) 88 ⁇ 1 (3) discodermolide (2) 76 ⁇ 6 (4) nd 90 ⁇ 1 (3) paclitaxel nd 6 ⁇ 5 (3) 26 + 1 (3)
• the mixture was stirred at 0 °C for 2 h and then quenched by the addition of saturated aqueous NaHCO 3 .
• the phases were separated and the aqueous layer was extracted with CH 2 C1 2 .
• the combined organic phases were washed with 0.5 M aqueous NaHSO 4 .
• the organic phase was dried over Na SO 4 , filtered and concentrated under reduced pressure.
• PPh 3 (7.05 g, 26.2 mmol), imidazole (1.78 g, 26.2 mmol), diisopropylethylamine (4.6 mL, 26.2 mmol) in benzene (80 mL), diethyl ether (165 mL) and acetonitrile (33 mL) were stirred at room temperature and treated with iodine (6.65 g, 26.2 mmol). The resulting mixture was vigorously stirred until the formation of a beige suspension. A solution of the alcohol 21 (5.0 g, 13.1 mmol) in Et O (20 mL) was added dropwise to the suspension and the resulting mixture was stirred at room temperature for 30 min.
• the resulting mixture was stined at -78 °C for 1 h, at 0 °C for 15 min and at room temperature for 5 min.
• the suspension was cooled to 0 °C and the iodide was added as a solution in THF (6 mL followed by a 6 mL rinse).
• the reaction mixture was stirred at room temperature for 24 h and quenched with half-saturated aqueous NH 4 C1.
• TBSOTf (0.40 mL, 1.74 mmol) was added to a stirred solution of alcohol 41 ⁇ (1.39 g, 1.17 mmol) and 2,6-lutidine (0.27 mL, 2.33 mmol) in CH 2 C1 2 (23 mL) at 0 °C. After stirring for 1 h at ambient temperature, the reaction mixture was quenched by the addition of water (50 mL) and extracted by CH 2 C1 .
• N O- dimethylhydroxylamine hydrochloride 0.064 g, 0.65 mmol
• Et 3 ⁇ 0.09 mL, 0.65 mmol
• DMAP 8 mg, 0.065 mmol
• the reaction mixture was cooled to 0 °C, DCC (0.14 g, 0.65 mmol) was added.
• the mixture was stirred at ambient temperature for 15 h and filtered.
• the filtrate was washed with 0.5 N HCI, saturated aqueous NaHCO 3 , and brine, dried over anhydrous MgSO 4 and concentrated.
• Ynone 82 (7.06 g, 7.59 mmol) was taken up in z ' -PrOH (100 mL). Noyori catalyst (1.02 g, 1.52 mmol, 20 mol%) was added in one portion and the solution was stirred for 12 h.
• ketophosphonate 38 (0.85 g, 2.20 mmol) and Ba(OH) 2 (0.30 g, activated by heating to 100 °C for 1-2 h before use) in THF (40 mL) was stirred at room temperature for 30 min.
• the reaction mixture was diluted with Et 2 O (30 mL) and washed with sat'd NaHCO 3 (50 mL) and brine (50 mL). The organic solution was dried (MgSO 4 ) and the solvent was evaporated in vacuo.
• NiCl 2 *6H 2 O (0.20 g, 0.84 mmol) then portionwise NaBH 4 (0.17 g, 4.49 mmol) were added to a stirred solution of unsaturated ketone 89 (2.60 g, 1.72 ⁇ mol) in MeOH (60 mL), THF (20 mL) at 0 °C. After 1 h, the reaction mixture was evaporated and filtered with celite using Et 2 O as an eluent (30 mL).
• TBSOTf (0.30 mL, 2.57 mmol) was added to a stirred solution of alcohol 91 ⁇ (1.02 g, 0.86 mmol) and 2,6-lutidine (0.20 mL, 1.71 mmol) in CH 2 C1 2 (17 mL) at 0 °C and the reaction mixture was stirred for 1 h at ambient temperature. The reaction mixture was quenched by the addition of water (50 mL). The reaction mixture was extracted by CH 2 C1 2 and dried over MgSO 4 followed by the evaporation of the solution under reduced pressure.
• reaction mixture was quenched by addition of a sat'd NH 4 C1 solution (5 mL) and diluted with diethyl ether (20 mL). The layers were separated and organic phase was washed with brine (30 mL) and dried with MgSO 4 , filtered, and concentrated.
• reaction mixture was quenched by adding sat'd NaHCO 3 (5 mL).
• the organic phase was washed by sat'd NaHCO 3 solution (3 x 10 mL) and brine, dried over MgSO 4 and concentrated.
• reaction mixture was stirred at 0 °C for 30 min and then added to 4-DMAP (60 mL, 0.02 M solution in toluene) at 25 °C and stirred overnight.
• the reaction mixture was concentrated, Et O (10 mL) was added and the crude was washed with 0.5 N HCI (2 x 10 mL), dried over MgSO .
• Tubulin polymerization assay Tubulin polymerization assay. Tubulin assembly was monitored turbidimetrically in Gilford 250 spectrophotometers equipped with electronic temperature controllers as described previously (ter Haar et al, 1996). The reaction mixtures without the compounds consisted of tubulin (1 mg/ml), heat-treated MAPs (0.75 mg/ml, if present), GTP (100 ⁇ M, if present), and 0.1M (4-morpholinyl)ethane sulfonate (MeS). Baselines were established after addition of all reaction components except the compounds to the cuvettes held at 0°C.
• GI 50 values The fifty percent growth inhibition values were calculated for the compounds against all the three cell lines.
• Pelleting Assay determination ofECso: The assay was performed under three different reaction conditions following the procedure reported earlier (Gapud et al, 2004).
• Reaction condition 1 included 0.2 M monosodium glutamate (MSG), 10 ⁇ M tubulin, 5% DMSO and varying concentrations of test agents.
• Reaction condition 2 included 0.8 M MSG, 400 ⁇ M GTP, 10 ⁇ M tubulin, 5% DMSO, and varying concentrations of test agents.
• Reaction condition 3 had 0.6 M MSG, 200 ⁇ M GTP, and 10 ⁇ M tubulin, and 5% DMSO, and varying concentrations of the test agents.
• the experimental protocol for all the three reaction conditions was the following.
• the reaction mixtures were incubated at room temperature (20-22 °C) for 15 min and spun for 10 min at 14,000 rpm in an Eppendorf microtube centrifuge. Aliquots of the supernatants were removed and assayed for protein content by the method of Lowry.
• the EC 0 was defined as drug concentration required to polymerize 50% of tubulin compared to the pellet found in the DMSO control reaction determined for each test system. On average 5.5 + 4.0% of the tubulin pelleted in the DMSO control.
• Multiparameter fluorescence microscopy high information content cell-based fluorescence screening HeLa cells growing at log phase were trypsinized and plated in 40 ⁇ L at a density of 7,000-8,000 cells per well in calf skin collagen I-coated 384-well plates (Falcon #3962; Fisher Scientific). Cells were exposed to test agents or 0.5% DMSO within 2- 8 h of plating. Concentrated DMSO stock solutions of all test agents were diluted into solutions of HBSS medium plus 10% FBS and added to the microplate wells (10 ⁇ L per well), using an automated liquid handling system (Biomek® 2000; Beckman-Coulter, Inc.) to provide a serial 2-fold dilution of each test agent.
• an automated liquid handling system Biomek® 2000; Beckman-Coulter, Inc.
• This step extracts a fraction of the soluble cellular components, including soluble tubulin.
• the wells were washed with HBSS (100 ⁇ L/well), followed by addition of a primary antibody solution containing mouse anti- ⁇ - tubulin (1:3000) and rabbit anti-phosphohistone H3 (1:500) in HBSS (10 ⁇ L/well). After 1 h at room temperature, the wells were washed with HBSS as above, followed by the addition of a secondary antibody solution containing fluorescein-5-isothiocyanate (FITC)-labeled donkey anti-mouse (1:300) and Cy3-labeled donkey anti-rabbit (1:300) antibodies diluted in HBSS (10 ⁇ L/well).
• FITC fluorescein-5-isothiocyanate
• HBSS HBSS was added (100 ⁇ L/well).
• the plates were placed in an ArrayScan® HCS Reader with the Target Activation BioApplication Software coupled to Cellomics® Store and the vHCSTM Discovery Toolbox (Cellomics, Inc.) to analyze images. Briefly, the instrument was used to scan multiple optical fields, each with multiparameter fluorescence, within a subset of the wells of the 384- well microplate.
• the Bio Application software produced multiple numerical feature values, such as subcellular object intensities, shapes, and location for each cell within an optical field. Data were acquired from a minimum of 1,000 cells per well, except in cases where added test agents reduced the attachment of cells to the substrate.

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