U.S. patent number RE46,965 [Application Number 13/924,892] was granted by the patent office on 2018-07-24 for intermediates for the preparation of analogs of halichondrin b.
This patent grant is currently assigned to Eisai R&D Management Co., Ltd.. The grantee listed for this patent is Eisai R&D Management Co., Ltd.. Invention is credited to Brian Austad, Trevor Calkins, Charles E. Chase, Francis G. Fang, Bryan M. Lewis.
United States Patent |
RE46,965 |
Austad , et al. |
July 24, 2018 |
**Please see images for:
( Certificate of Correction ) ** |
Intermediates for the preparation of analogs of Halichondrin B
Abstract
The present invention provides macrocyclic compounds, synthesis
of the same and intermediates thereto. Such compounds, and
compositions thereof, are useful for treating or preventing
proliferative disorders Formula (F-4). ##STR00001##
Inventors: |
Austad; Brian (Tewksbury,
MA), Chase; Charles E. (Londonderry, NH), Fang; Francis
G. (Andover, MA), Calkins; Trevor (Stoughton, WI),
Lewis; Bryan M. (North Brunswick, NJ) |
Applicant: |
Name |
City |
State |
Country |
Type |
Eisai R&D Management Co., Ltd. |
Tokyo |
N/A |
JP |
|
|
Assignee: |
Eisai R&D Management Co.,
Ltd. (Tokyo, JP)
|
Family
ID: |
34972515 |
Appl.
No.: |
13/924,892 |
Filed: |
June 24, 2013 |
PCT
Filed: |
June 03, 2005 |
PCT No.: |
PCT/US2005/019669 |
371(c)(1),(2),(4) Date: |
March 28, 2007 |
PCT
Pub. No.: |
WO2005/118565 |
PCT
Pub. Date: |
December 15, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
13495909 |
Jan 6, 2015 |
RE45324 |
|
|
|
60576642 |
Jun 3, 2004 |
|
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60626769 |
Nov 10, 2004 |
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60663300 |
Mar 18, 2005 |
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Reissue of: |
11628396 |
Jun 3, 2005 |
7982060 |
Jul 19, 2011 |
|
Reissue of: |
11628396 |
Jun 3, 2005 |
PCT/US2005/019669 |
Jun 3, 2005 |
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
C07H
15/26 (20130101); C07D 493/22 (20130101); C07D
307/12 (20130101); C07D 407/14 (20130101); C07D
493/22 (20130101); A61P 35/00 (20180101); C07D
307/20 (20130101); C07C 33/423 (20130101); C07D
407/06 (20130101); C07D 493/10 (20130101); C07D
307/12 (20130101); C07D 493/04 (20130101); C07F
7/188 (20130101); A61K 31/341 (20130101); C07D
493/18 (20130101); C07D 493/20 (20130101); C07D
307/33 (20130101); A61P 35/02 (20180101); C07D
493/08 (20130101); C07F 7/1804 (20130101); C07D
307/28 (20130101); C07D 493/08 (20130101); A61K
31/341 (20130101); C07F 7/188 (20130101); C07F
7/1804 (20130101); C07D 493/04 (20130101); C07D
493/18 (20130101); C07D 317/72 (20130101); C07D
493/10 (20130101); C07C 33/423 (20130101); C07D
407/06 (20130101); C07D 307/33 (20130101); C07D
493/20 (20130101); C07D 307/28 (20130101); C07H
15/26 (20130101); C07D 407/14 (20130101); C07D
317/72 (20130101); Y02P 20/55 (20151101); Y02P
20/55 (20151101) |
Current International
Class: |
C07D
307/12 (20060101); C07D 493/04 (20060101); C07D
407/14 (20060101); C07D 407/06 (20060101); C07F
7/18 (20060101); C07D 493/08 (20060101); C07D
307/33 (20060101); C07D 307/28 (20060101); C07D
493/18 (20060101); C07D 493/22 (20060101); C07D
493/20 (20060101); C07D 317/72 (20060101); C07H
15/26 (20060101); C07C 33/42 (20060101); A61K
31/341 (20060101); C07D 493/10 (20060101) |
Field of
Search: |
;514/450 ;549/348 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
|
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|
|
|
0 572 109 |
|
Dec 1993 |
|
EP |
|
0572109 |
|
Dec 1993 |
|
EP |
|
0572109 |
|
Dec 1993 |
|
EP |
|
2002-518384 |
|
Jun 2002 |
|
JP |
|
2002-518384 |
|
Jun 2002 |
|
JP |
|
WO 93/17690 |
|
Sep 1993 |
|
WO |
|
WO-93/17690 |
|
Sep 1993 |
|
WO |
|
WO 99/65894 |
|
Dec 1999 |
|
WO |
|
WO-99/65894 |
|
Dec 1999 |
|
WO |
|
WO 2004034990 |
|
Apr 2004 |
|
WO |
|
WO 2005/118565 |
|
Dec 2005 |
|
WO |
|
WO 2005118565 |
|
Dec 2005 |
|
WO |
|
WO 2006076100 |
|
Jul 2006 |
|
WO |
|
WO 2007061874 |
|
May 2007 |
|
WO |
|
WO 2009/046308 |
|
Apr 2009 |
|
WO |
|
WO-2009/046308 |
|
Apr 2009 |
|
WO |
|
WO 2009/064029 |
|
May 2009 |
|
WO |
|
WO-2009/064029 |
|
May 2009 |
|
WO |
|
WO 2009/124237 |
|
Oct 2009 |
|
WO |
|
WO-2009/124237 |
|
Oct 2009 |
|
WO |
|
WO 2011/094339 |
|
Aug 2011 |
|
WO |
|
WO-2011/094339 |
|
Aug 2011 |
|
WO |
|
WO 2012147900 |
|
Nov 2012 |
|
WO |
|
WO 2014087230 |
|
Jun 2014 |
|
WO |
|
WO 2015066729 |
|
May 2015 |
|
WO |
|
WO 2015085193 |
|
Jun 2015 |
|
WO |
|
WO 2015134399 |
|
Sep 2015 |
|
WO |
|
WO 2015183961 |
|
Dec 2015 |
|
WO |
|
WO 2015184145 |
|
Dec 2015 |
|
WO |
|
WO 2016141209 |
|
Sep 2016 |
|
WO |
|
WO 2016179607 |
|
Nov 2016 |
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WO |
|
Other References
Duan and Kishi, "Synthetic Studies on Halichondrins: A New
Practical Synthesis of the C.1-C.12 Segment," Tetrahedron Lett.
34(47), pp. 7541-7544 (1993). cited by applicant .
Stamos et al., "A Mild Preparation of Vinyliodides from
Vinylsilanes," Tetrahedron Lett. 37(48), pp. 8647-8650 (1996).
cited by applicant .
Communication pursuant to Rule 114(2) EPC from counterpart European
Application EP 05760356.5, dated Jul. 4, 2012. cited by applicant
.
Wan et al., "Asymmetric Ni(II)/Cr(II)-Mediated Coupling Reaction:
Stoichiometric Process," Organic Letters 4(25), pp. 4431-4434
(2002). cited by applicant .
Supporting Information for Wan et al., "Asymmetric
Ni(II)/Cr(II)-Mediated Coupling Reaction: Stoichiometric Process,"
Organic Letters 4(25), pp. 4431-4434 (2002). cited by applicant
.
RN: 546141-26-8; CN: 1,2-Propaneldiol,
3-[(2R,3R,4R,5S)-tetrahydro-3-hydroxy-4-[(phenylsulfony)methyl]-5-(2-prop-
en-1-yl)-2-furanyl]-1,2-dibenzoate, (2S)-; Entered STN: Jul. 11,
2003. cited by applicant .
RN: 546141-39-3; CN: 1,2-Propaneldiol,
3-[(2R,3R,4R,5S)-tetrahydro-3-methoxy-4-[(phenylsulfony)methyl]-5-(2-prop-
en-1-yl)-2-furanyl]-,1,2-dibenzoate, (2S); Entered STN: Jul. 11,
2003. cited by applicant .
RN: 546141-40-6; CN: 1,2-Propaneldiol,
3-[(2R,3R,4R,5S)-tetrahydro-3-methoxy-4-[(phenylsulfony)methyl]-5-(2-prop-
en-1-yl)-2-furanyl]-, (2S); Entered STN: Jul. 11, 2003. cited by
applicant .
RN: 185411-09-0; CN: L-arabino-D-allo-Dodeconic acid, 3,7:6,
10-dianhydro-8,9-O-cyclohexylidene-2,4,5-trideoxy-, methyl ester;
Entered STN: Jan. 28, 1997. cited by applicant .
English translation of the Office Action from the Japanese Patent
Office for the counterpart Japanese Application No. 2007-515,643,
dated Apr. 24, 2012. cited by applicant .
English translation of the Office Action from the Japanese Patent
Office for the counterpart Japanese Application No. 2007-515,643,
dated Aug. 16, 2012. cited by applicant .
Takai et al., "Reactions of Alkenylchromium reagents prepared from
alkenyl trifluoromethanesulfonates (triflates) with chromium (II)
chloride under nickel catalysis," J. Am. Chem. Soc. 108, pp.
6048-6050 (1986). cited by applicant .
Third Party Observation pursuant to Article 115 EPC relating to
counterpart European Application EP 05760356.5, dated Mar. 26,
2012. cited by applicant .
Third Party Observation pursuant to Article 115 EPC relating to
counterpart European Application EP 05760356.5, dated Jan. 9, 2013.
cited by applicant .
U.S. Patent and Trademark Office, "Clarification of Criteria for
Reissue Error in View of In re Tanaka " dated Aug. 1, 2011. cited
by applicant .
File History of U.S. Appl. No. 13/868,641, "Intermediates for the
Preparation of Analogs of Halichondrin B," in the name of Farid
Benayoud et al., filed Apr. 23, 2013. cited by applicant .
Aicher, T.D., et al.; "Synthetic Studies Towards Halichondrins:
Synthesis of the C.27-C.38 Segment," Tetrahedron Lett. 33(12):
1549-1552 (1992). cited by applicant .
Austad et al,, "Process development of Halaven.RTM.: Synthesis of
the C14-C35 fragment via iterative Nozaki-Hiyama-Kishi
reaction--Williamson ether cyclization," Synlett. 24:327-332
(2013). cited by applicant .
Austad et al., "Commercial manufacture of Halaven.RTM.:
Chemoselective transformations en route to structurally complex
macrocyclic ketones," Synlett. 24;333-337 (2013). cited by
applicant .
Austad et al., "Commercial manufacture of Halaven.RTM.:
Chemoselective transformations en route to structurally complex
macrocyclic ketones," Synlett. 24 (2013). Supporting Information,
13 pages. cited by applicant .
Bai et al., "Halichondrin B and Homohalichondrin B, Marine Natural
Products Binding in the Vinca Domain to Tubulin. Discovery of
Tubulin-based Mechanism of Action by Analysis of Differential
Cytotoxicity Data," J. Biol. Chem. 266(24): 15882-15889 (1991).
cited by applicant .
Burke, S.D., et al., "Enantioselective Synthesis of a Halichondrin
B C(20) 'C(36) Precursor," Tetrahedron Lett., 36(39): 7023-7026
(1995). cited by applicant .
Burke; S.D., et al., "Synthesis of a C(22)-C(34) Halichondrin B
Precursor via Ring Opening--Double Ring Closing Metathesis." J.
Org. Chem., 63: 8626-8627 (1998). cited by applicant .
Burke, S.D., et al., "Synthesis of a C(22) ' C(34) Halichondrin
Precursor via a Double Dioxanone-to-Dihydropyran Rearrangement,"
Tetrahedron Lett., 32(32): 3961-3964 (1991 ). cited by applicant
.
Aicher et al., "Total Synthesis of Halichondrin Band
Norhalichondrin B," J. Am. Chem. Soc. 114(8): 3162-3164 (1992).
cited by applicant .
Chen C., et al., "Ni(II)/Cr(II)-Mediated Coupling Reaction: An
Asymmetric Process," J. Org. Chem., 60:5386-5387 (1995). cited by
applicant .
Chase et al., "Process development of Halaven.RTM.: Synthesis of
the Cl1-C13 fragment from D-(-)-Gulono-1, 4-lactone," Synlett.
24:323-326 (2013). cited by applicant .
Choi et al., "Assymrnetric Ni(II)/Cr(II)-Mediated Coupling
Reaction: Catalytic Process," Org Lett. 4(25): 4435-4438 (2002).
cited by applicant .
Cooper, A.J., et al., "Total Synthesis of Halichondrin B from
Common Sugars: An F-Ring Intermediate from D-Glucose and Efficient
Construction of the C1 to C21 Segment," Tetrahedron Lett., 34(51):
8193-8196 (1993). cited by applicant .
Dong, C. et al. "New Syntheses of E7389 C14 ?C35 and Halichondrin
C14 ?C38 Building Blocks: Reductive Cyclization and Oxy-Michael
Cyclization Approaches" J. Am. Chem. Soc. 131: 15642-15646 (2009).
cited by applicant .
Duan and Kishi, "Synthetic studies on halichondrins: A new
practical synthesis of the C.1-C.12 segment," Tetrahedron Lett.
34(47):7541-7544 (1993). cited by applicant .
Flemming et al., "Nitrile Anion Cyclizations," Tetrahedron 58:1-23
(2002). cited by applicant .
Hirata et al., "Halchondrins--Antitumor Polyether Macrolides from a
Marine Sponge," Pure Appl. Chem. 58(5): 701-710 (1986). cited by
applicant .
Horita et al., "Synthetic Studies of Halichondrin B, an Antitumor
Polyether Macrolide Isolated from a Marine Sponge. 8. Synthesis of
the Lactone Part (C1-C36) via Horner-Emmons Coupling Between C1-C15
and C16-C36 Fragments and Yamaguchi Lactonization," Tetrahedron
Lett, 38(52): 8965-8968 ( 1997). cited by applicant .
Horita, K., et al., "Research on Anti-Tumor Active Site of Marine
Source Natural Product, Halichondrin B.," International Congress
Series, 1157 (Towards Natural Medicine Research in the 21st
Century), 327-336 (1998). cited by applicant .
Horita, K., et al., "Synthetic Studies of Halichondrin B, an
Antitumor Polyether Macrolide Isolated From a Marine Sponge, 2.
Efficient Synthesis of C 16-C26 Fragments via Construction of the D
Ring by a Highly Stereocontrolled Iodoetherification," Synlett,
40-43 ( 1994 ). cited by applicant .
Horita, K., et al., "Synthetic Studies of Halichondrin B, an
Antitumor Polyether Macrolide Isolated from a Marine Sponge, 3.
Synthesis of C27-C36 Subunit via Completely Stereoselective
C-Glycosylation to the F ring," Synlett, 43-45 (1994). cited by
applicant .
Horita, K., et al., "Synthetic Studies of Halichondrin B. an
Antitumor Polyether Macrolide Isolated from a Marine Sponge. 7.
Synthesis of Two C27-C36 Units via Construction of the F ring and
Completely Stereoselective C-Glycosylation Using Mixed Lewis
Acids," Chem. Pharm. Bull., 45(10): 1558-1572 (1997). cited by
applicant .
Horita, K., et al., "Synthetic Study of a Highly Antitumorigenic
Marine Phytochemical, Halichondrin B," Phytochemicals and
Phytopharmaceuticals, Shahihi, F. and Ho, C.-T., Eds., AOCS Press,
Champaign, IL, 2000. 386-397. cited by applicant .
Jackson et al.. "A Total Synthesis of Norhalichondrin B" Angew.
Chem. Int. Ed. 48: 2346-2350 (2009). cited by applicant .
Jiang, L., et al., "A Novel Route to the F-Ring of Halichondrin B.
Diastereoselection in Pd(O)-Mediated meso and C2 Diol
Desymmetrization," Org. Lett., 4(20): 3411-3414 (2002). cited by
applicant .
Jiang, L., et al., "A Practical Synthesis of the F-Ring of
Halichondrin B via Ozonolytic Desymmetrization of a C2-Symmetric
Dihydroxycyclohexene," J. Org. Chem., 68: 1150-1153 (2003). cited
by applicant .
Kim, D. et al. "New Syntheses of E7389 C14?C35 and Halichondrin
C14?C38 Building Blocks: Double-Inversion Approach" J. Am. Chem.
Soc. 131: 15636-15641 (2009). cited by applicant .
Mattocks, "Novel Reactions of Some ?-Acyloxy Acid Chlorides," J.
Chem. Soc. 371: 1918-1930 (1964). cited by applicant .
Mattocks, "Novel Reactions of Some ?-Acyloxy-acid Halides," J.
Chem. Soc. 932: 4840-4845 (1964). cited by applicant .
Newman, "Drug Evaluation: Eribulin, a Simplified Ketone Analog of
the Tubulin Inhibitor Halichondrin B, for the Potential Treatment
of Cancer," Curr. Opin. Invest. Drugs, 8:1057-1066 (2007). cited by
applicant .
Sakamoto et al., "Stereoselective Ring Expansion via Bicyclooxonium
ion. A Novel Approach to Oxocanes," Org. Lett. 4(5):676-678 (2002).
cited by applicant .
Schreiber, "Hydrogen Transfer from Tertiary Amines to
Trifluorcacetic Anhydride." Tetrahedron Lett. 21:1027-1030 (1980).
cited by applicant .
Stamos et al., "A mild preparation of vinyliodides from
vinylsilanes," Tetrahedron Lett. 37(48): 8647-8650 (1996). cited by
applicant .
Stamos et al., "New Synthetic Route to the C.14-C.38 Segment of
Halichondrins," J. Org. Chem. 62:7552-7553(1997). cited by
applicant .
Stamos et al., "Synthetic Studies on Halichondrins: A Practical
Synthesis of the C.1-C.13 Segment" Tetrahedron Lett. 37(48):
8643-8646 (1996). cited by applicant .
Stamos, D.P., et al., "Ni(II)/Cr(II)-Mediated Coupling Reaction:
Beneficial Effects of 4-Tert-Butylpyridine as an Additive and
Development of New and Improved Workup Procedures," Tetrahedron
Lett., 38(36): 6355-6358 (1997). cited by applicant .
Towle et al. "Halichondrin B Macrocyclic Ketone Analog E7389:
Medicinal Chemistry Repair of Lactone Ester Instability Generated
During Structural Simplification to Clinical Candidate" Annual
Meeting of the American Association for Cancer Research, Apr. 6-1
0. 2002, 5721. cited by applicant .
Vahdat et al., "Phase II Study of Eribulin Mesylate, a Halichondrin
B Analog, in Patients with Metastatic Breast Cancer Previously
Treated with an Anthracycline and a Taxane," J. Clin. Oncol.
27(18): 2964-2961 (2009). cited by applicant .
Wan et al., "Asymmetric Ni(II)/Cr(II)-Mediated Coupling Reaction:
Stoichiometric Process," Org. Lett. 4(25): 4431-4434 (2002). cited
by applicant .
Xie, C., et al., "Synthesis of the C20-C26 Building Block of
Halichondrins via a Regiospecific and Stereoselective SN2'
Reaction," Org. Lett., 4(25):4427-4429 (2002). cited by applicant
.
Yang et al., "Second Generation Synthesis of C27-C35 Building Block
of E7389, a Synthetic Halichondrin Analogue," Org. Lett. 11 (20):
4516-4519 (2009). cited by applicant .
Yu et al., New Synthetic Route to the C.14-C.21 Fragment of
Halichondrin B, Book of Abstracts, 219th ACS National Meeting, San
Francisco, CA, Mar. 26-30, 2000 (2000). cited by applicant .
RN 1 85411-09-0 CN L-arabino-D-allo-Dodeconic acid, 3,7:6, 1
0-dianhydro-8,9-0-cyclohaxylidene-2,4,5-trideoxy-, methyl ester
Entry Date: Entered STN: Jan. 28, 1997. cited by applicant .
RN 546141-26-8 CN 1 ,2 Propanediol,
3-[(2R,3R,4R,5S)-tetrahydro-3-hydroxy-4-[(phenylsulfonyl)methyl]-5-(2-pro-
pen-1-yl)-2-furanyl]-, 1,2-dibenzoate, (2S)--Entry Date: Entered
STN: Jul. 11, 2003. cited by applicant .
RN 546141-39-3 CN 1 ,2-Propanediol,
3-[(2R,3R,4S,5S)-tetrahydro-3-methoxy-4-[(phenylsulfonyl)methyl]-5-(2-pro-
pen-1-yl)-2-furanyl]-, 1,2-dibenzoate, (2S)--Entry Date: Entered
STN: Jul. 11, 2003. cited by applicant .
RN 546141-40-6 CN 1 ,2-propanediol,
3-[(2R,3R,4S,5S)-tetrahydro-3-methoxy-4-f(phenylsulfonyl)methyl]-5-(2-pro-
pen-14)-2-furanvll-, (2S)--ED Entered STN: Jul. 11, 2003. cited by
applicant .
Wan et al., "Asymmetric Ni(II)/Cr(II)-Mediated Coupling Reaction:
Stoichiometric Process," Org. Lett. 4(25): 4431-4434 (2002)
Supporting Information, 8 pages. cited by applicant .
Written Opinion from International Application No.
PCT/US2005/019669, dated Sep. 7, 2005. cited by applicant .
Takai et at "Reactions of Alkenylchromium Reagents Prepared from
Alkenyl Trifluoromethanesulfonates (Triflates) with Chromium(l1)
Chloride under Nickel Catalysis" J. Am. Chem. Soc. 1966, 108:6048.
cited by applicant .
Communication pursuant to Rule 114(2) EPC from counterpart European
Application EP 05760356.5, dated Apr. 3, 2012. cited by applicant
.
Office Action from the Canadian Intellectual Property Office for
the counterpart Canadian Application No. 2,567,984, dated Oct. 7,
2011. cited by applicant .
English translation of the Office Action from the Japanese Patent
Office for the counterpart Japanese Application No. 2007-515,643,
dated Sep. 13, 2011. cited by applicant .
English translation of the Office Action from the Chinese Patent
Office for the counterpart Chinese Application No. 201010236637.2,
dated Sep. 16, 2011. cited by applicant .
Jerry March, "Advanced Organic Chemistry," 4th Ed., pp. 348-357,
John Wiley and Sons, N.Y. (1992). cited by applicant .
English translation of the Office Action from the Chinese Patent
Office for the counterpart Chinese Application No. 201010236637.2,
dated Jul. 4. 2012. cited by applicant .
Communication enclosing the Extended European Search Report for
European Patent Application No. 12178696,6, dated Oct. 16. 2012.
cited by applicant .
Sutherland et al., "The Synthesis of 6.alpha.- and
6.beta.-Fluoroshikimic Acids," J. Chem. Soc., Chem. Commun.
18:1386-1387, 1989. cited by applicant .
Greene et al., "Protective Groups in Organic Synthesis," John Wiley
& Sons, Inc., New York, Third Edition, pp. 24, 127, 128, 134,
142, 170,207,209,215, and 216 (1999). cited by applicant .
March, "Advanced Organic Chemistry," John Wiley & Sons, New
York, Fourth Edition, pp. 386-388 (1992). cited by applicant .
Carruthers et al., "Modern Methods of Organic Synthesis," Cambridge
University Press, Cambridge, Fourth Edition, p. 65 (2004). cited by
applicant .
Ritter, "Synthetic Transformations of Vinyl and Aryl Triflates,"
Synthesis 8:735-762, 1993. cited by applicant .
Nicolaou et al., "Total Synthesis of the CP Moiecules CP-263, 114
and CP-225,917--Part 1. Synthesis of Key Intermediates and
Intelligence Gathering," Angew. Chem. Int. Ed. 38:1669-1675, 1999.
cited by applicant .
Nicolaou et al., "Total Synthesis of Brevetoxin A: Part 3:
Construction of GHIJ and BCDE Ring Systems," Chem. Eur. J.
5:628-645, 1999. cited by applicant .
The AkzoNobel Technical Bulletin. "Diisobutylaluminum hydride
(DIBAL-H) and Other Isobutyl Aluminum Alkyls (DIBAL-BOT, TIBAL) as
Specialty Organic Synthesis Reagents," 14 pages (2006). cited by
applicant .
Wang et al., "Facile preparation of peracetates and
per-3-bromobenzoates of alpha-mono- and disaccharides," Molecules 1
0:1325-1334, 2005. cited by applicant .
Youssefyeh, "Acylatrons of Ketals and Errol Ethers,"J. Am. Chem.
Soc., 85:3901-3902, 1963. cited by applicant .
Ph.D. Thesis of Thomas Daniel Aicher, "Synthetic Studies towards
Halichondrin B," Chapter 4, pp. 35-54, 1990. cited by applicant
.
First Examination Report from the India Patent Office for the
counterpart India Application No. 4812/CHENP/2006 dated Oct. 12,
2012. cited by applicant .
Written Amendment and Written Argument filed in the Japanese Patent
Office for the counterpart Japanese Application No.
2007-515,643dated Feb. 4, 2013. cited by applicant .
Communication pursuant to Article 94(3)PEC from the European Patent
Office for the counterpart EP Application No. 12178696.6 dated Aug.
8, 2013. cited by applicant .
Notification of Grounds for Rejection from the South Korea Patent
Office for the counterpart South Korean Application No.
10-2007-7000085 dated Jun. 30, 2013. cited by applicant .
Response to Office Action filed in the European Patent Office for
the counterpart EP Application No. 05760356.5 dated Sep. 12, 2013,
cited by applicant .
Result of Consultation from the European Patent Office for the
counterpart EP Application No. 05760356.5 dated Oct. 16, 2013.
cited by applicant .
Written Submissions filed in the from the European Patent Office
for the counterpart EP Application No. 05760356.5 dated Oct. 21,
2013. cited by applicant .
Notice of Oral Proceedings from the European Patent Office
scheduling Oral Proceedings on Nov. 21, 2013, for the counterpart
EP Application No. 05760356.5 dated Oct. 29, 2013. cited by
applicant .
Supporting Information for Choi, Hyeong-Wook, et al., Asymmetric
Ni(II)/Cr(II)-Mediated Coupling Reaction: CaLatytic Process,
Organic Letters, vol. 4, No. 25, pp. 1-8 (2002). cited by applicant
.
Namba, K., et al., "New Catalytic Cycle for Couplings of Aldehydes
with Organochromium Reagents", Organic Letters., vol. 6, No. 26,
pp. 5031-5033 (2004). cited by applicant .
Supporting information for Kurosu, M., et al., "Fe/Cr- and
Co/Cr-Mediated Catalytic Asymmetric 2-Haloallylations of
Aldehydes", Journal of the American Chemical Society, vol. 126, No.
39, pp. S-1-5-9, (2004). cited by applicant .
Communication enclosing the extended European search report for
European Patent Application No. 15159875.2, dated Oct. 12, 2015 (7
pages). cited by applicant .
Choi et al., "Synthetic Studies on the Marine Natural Product
Halichondrins," Pure Appl. Chem. 75:1-17, 2003. cited by applicant
.
International Preliminary Report on Patentability from
International Application No. PCT/US2005/019669, issued Dec. 4,
2006. cited by applicant .
International Search Report from International Application No.
PCT/US2005/019669 dated Aug. 29, 2005 (date of completion of
search) and Sep. 7, 2005 (date of mailing of report). cited by
applicant .
Written Opinion from International Application No.
PCT/US2005/019669, mailed Sep. 7, 2005. cited by applicant .
Anderson, "Developing Processes for Crystallization-Induced
Asymmetric Transformation," Org. Process. Res. Dev. 9: 800-813
(2005). cited by applicant .
Bernet et al., "Carbocyclische Verbindungen aus Monosacchariden.
Umsetzungen in der Glucosereihe," Helv. Chim. Acta. 62: 1990-2016
(1979). cited by applicant .
Blanchette et al., "Horner-Wadsworth-Emmons Reaction: Use of
Lithium Chloride and an Amine for Base-Sensitive Compounds,"
Tetrahedron Lett. 25(21): 2183-2186 (1984). cited by applicant
.
Burke, S.D., et al., "Synthetic Studies Toward Complex Polyether
Macrolides of Marine Origin," Spec. Publ. R. Soc. Chem., 198:
(Anti-Infectives), 73-85 (1997). cited by applicant .
Choi et al., "Synthetic Studies on the Marine Natural Product
Halichondrins," Pure Appl. Chem. 75(1): 1-17 (2003). cited by
applicant .
Hirata et al., "Halichondrins--Antitumor Polyether Macrolides from
a Marine Sponge," Pure Appl. Chem. 58(5): 701-710 (1986). cited by
applicant .
Hori et al., "Efficient Synthesis of 2,3-trans-Tetrahydropyrans and
Oxepanes: Rearrangement-Ring Expansion of Cyclic Ethers Having a
Chloromethanesulfonate," Tetrahedron Lett. 40: 2145-2148 (1999).
cited by applicant .
Horita et al., "Synthetic Studies of Halichondrin B, an Antitumor
Polymer Macrolide Isolated from a Marine Sponge. 8. Synthesis of
the Lactone Part (C1-C36) via Horner-Emmons Coupling Between C1-C15
and C16-C36 Fragments and Yamaguchi Lactonization," Tetrahedron
Lett. 38(52): 8965-8968 (1997). cited by applicant .
Horita, K., et al., "Synthetic Studies on Halichondrin B, an
Antitumor Polyether Macrolide Isolated from a Marine Sponge. 9.
Synthesis of the C16-C36 unit via Stereoselective Construction of
the D and E Rings," Chem. Pharm. Bull., 46(8): 1199-1216 (1998).
cited by applicant .
Jackson et al., "The Halichondrins and E7389," Chem. Rev. 109:
3044-3079 (2009). cited by applicant .
Kurosu et al., "Fe/Cr- and Co/Cr-Mediated Catalytic Asymmetric
2-Haloallylations of Aldehydes," J. Am. Chem. Soc. 126: 12248-12249
(2004). cited by applicant .
Mitsunobu, "The Use of Diethyl Azodicarboxylate and
Triphenylphosphine in Synthesis and Transformation of Natural
Products," Synthesis. 1-28 (1981). cited by applicant .
Tokunaga et al., "Asymmetric Catalysis with Water: Efficient
Kinetic Resolution of Terminal Epoxides by Means of Catalytic
Hydrolysis," Science 277: 936-938 (1997). cited by applicant .
Uemura et al., "Norhalichondrin A: An Antitumor Polyether Macrolide
from a Marine Sponge," J. Am. Chem. Soc. 107: 4796-4798 (1985).
cited by applicant .
Yu et al., Anticancer Agents from Natural Products; CRC Press: Boca
Raton, FL, 241-265. (2005). cited by applicant .
Zheng et al., "Macrocyclic Ketone Analogues of Halichondrin B,"
Bioorg. Med. Chem. Lett. 14: 5551-5554 (2004). cited by applicant
.
Zheng, W. et al. "Synthetic macrocyclic ketone analogs of
halichondrin B: structure-activity relationships" American
Association for Cancer Research, San Francisco, CA Apr. 1-5, 2000,
1915. cited by applicant .
Alley et al. "Comparison of the Relative Efficacies and Toxicities
of Halichondrin B Analogues" Proceedings of the AACR-NCI-EORTC
Conference on Molecular Targets and Cancer Therapeutics, Nov.
14-18, 2005, C230, p. 257. cited by applicant .
Dabybeen et al. "Comparison of the Activities of the Truncated
Halichondrin B Analog NSC 707389 (E7389) with Those of the Parent
Compound and a Proposed Binding Site on Tubulin" Molecular
Pharmacology 2006, 70:1866. cited by applicant .
Seletsky et al. "Structurally simplified macrolactone analogues of
halichondrin B" Bioorg. Med. Chem. Lett. 14:5547 (2004). cited by
applicant .
Towle et al. "In Vitro and In Vivo Anticancer Activities of
Synthetic Macrocyclic Ketone Analogues of Halichondrin B" Cancer
Research 2001, 61:1013. cited by applicant .
Wang et al. "Structure-Activity Relationships of Halichondrin B
Analogues: Modifications at C.30-C.38" Bioorg. Med. Chem. Lett.
2000, 10:1029. cited by applicant.
|
Primary Examiner: Diamond; Alan
Attorney, Agent or Firm: Finnegan, Henderson, Farabow,
Garrett & Dunner LLP
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
.[.This application claims priority under 35 U.S.C. .sctn.371 from
international application PCT/US05/019669, filed Jun. 3, 2005,
which claims priority to U.S. Provisional Patent Applications
60/576,642, filed Jun. 3, 2004, 60/626,769, filed Nov. 10, 2004,
and 60/663,300, filed Mar. 18, 2005, the entire contents of each of
which are hereby incorporated herein by reference..]. .Iadd.Notice:
More than one reissue application has been filed for the reissue of
U.S. Pat. No. 7,982,060. The reissue applications are Reissue
application. Ser. No. 13/495,909, and the present application filed
herewith. This application is a divisional reissue application of
U.S. patent application Ser. No. 13/495,909, filed Jun. 13, 2012,
which is a broadening reissue of U.S. Pat. No. 7,982,960, issued on
Jul. 19, 2011, from U.S. patent application Ser. No. 11/628,396,
which is the National Stage Entry of International Application No.
PCT/US2005/019669, filed Jun. 3, 2005, which claims the benefit of
priority of U.S. Provisional Application No. 60/576,642, filed on
Jun. 3, 2004, U.S. Provisional Application No. 60/626,769, filed on
Nov. 10, 2004, and U.S. Provisional Application No. 60/663,300,
filed on Mar. 18, 2005, all of which are incorporated herein by
reference in their entirety..Iaddend.
Claims
We claim:
.[.1. A compound of formula F-4: ##STR00154## wherein: each of
PG.sup.1, PG.sup.2, and PG.sup.3 is independently hydrogen or a
suitable hydroxyl protecting group; R.sup.1 is R or --OR; each R is
independently hydrogen, C.sub.1-4 haloaliphatic, benzyl, or
C.sub.1-4 aliphatic; and LG.sup.1 is a suitable leaving
group..].
.[.2. The compound according to claim 1, wherein said compound is
of formula F-4': ##STR00155## .].
.[.3. The compound according to claim 2, wherein R.sup.1 is OR
wherein R is hydrogen, methyl, or benzyl..].
.[.4. The compound according to claim 2, wherein PG.sup.1 and
PG.sup.2 are both hydrogen..].
.[.5. The compound according to claim 2, wherein each of PG.sup.1
and PG.sup.2 is independently a suitable hydroxyl protecting
group..].
.[.6. The compound according to claim 5, wherein one or both of
PG.sup.1 and PG.sup.2, taken together with the oxygen atom to which
each is bound, is a silyl ether or an arylalkyl ether..].
.[.7. The compound according to claim 6, wherein PG.sup.1 and
PG.sup.2 are both t-butyldimethylsilyl..].
.[.8. The compound according to claim 5, wherein PG.sup.1 and
PG.sup.2 are taken together, with the oxygen atoms to which they
are bound, to form a diol protecting group..].
.[.9. The compound according to claim 8, wherein said diol
protecting group is a cyclic acetal or ketal, a silylene
derivative, a cyclic carbonate, or a cyclic boronate..].
.[.10. The compound according to claim 2, wherein LG.sup.1 is
sulphonyloxy, optionally substituted alkylsulphonyloxy, optionally
substituted alkenylsulfonyloxy, optionally substituted
arylsulfonyloxy, or halogen..].
.[.11. The compound according to claim 10, wherein LG.sup.1 is
mesyloxy tosyloxy, chloro, iodo, bromo, or triflate..].
.[.12. The compound according to claim 2, wherein said compound is:
##STR00156## .].
.[.13. The compound according to claim 1, wherein said suitable
hydroxyl protecting group, taken with the oxygen atom to which it
is bound, is selected from an ester, an ether, a silyl ether, an
alkyl ether, an arylalkyl ether, and an alkoxyalkyl ether..].
.[.14. The compound according to claim 13, wherein said ester is a
formate, acetate, carbonate, or sulfonate; said silyl ether is a
trialkylsilyl ether; or said alkoxyalkyl ether is an acetal..].
.[.15. The compound according to claim 13, wherein said ester is
formate, benzoyl formate, chloroacetate, trifluoroacetate,
methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate,
3-phenylpropionate, 4-oxopentanoate,
4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetyl),
crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,
2,4,6-trimethylbenzoate, methyl carbonate, 9-fluorenylmethyl
carbonate, ethyl carbonate, 2,2,2-trichloroethyl carbonate,
2-(trimethylsilyl)ethyl carbonate, 2-(phenylsulfonyl)ethyl
carbonate, vinyl carbonate, ally! carbonate, or p-nitrobenzyl
carbonate; said silyl ether is trimethylsilyl, triethylsilyl,
t-butyldimethylsilyl, t-butyldiphenylsilyl, or triisopropylsilyl
ether; said alkyl ether is methyl, trityl, t-butyl, allyl, or
allyloxycarbonyl ether; said alkoxyalkyl ether is methoxymethyl,
methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl,
beta-(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ethers; or
said arylalkyl ether is benzyl, p-methoxybenzyl (MPM),
3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,
2,6-dichlorobenzyl, p-cyanobenzyl, 2- picolyl, or 4-picolyl..].
.[.16. The compound according to claim 1, wherein LG.sup.1 is
mesyloxy or tosyloxy..].
.[.17. The compound according to claim 9, wherein said cyclic
acetal or ketal is methylene, ethylidene, benzylidene,
isopropylidene, cyclohexylidene, or cyclopentylidene; or said
silylene derivative is di-t-butylsilylene or a
1,1,3,3-tetraisopropyldisiloxanylidene derivative..].
.[.18. The compound according to claim 1, wherein LG.sup.1 is
sulphonyloxy, optionally substituted alkylsulphonyloxy, optionally
substituted alkenylsulfonyloxy, optionally substituted
arylsulfonyloxy, or halogen..].
.Iadd.19. A compound of formula: ##STR00157## and/or a
pharmaceutically acceptable salt thereof..Iaddend.
.Iadd.20. A composition comprising a compound: ##STR00158## and/or
a pharmaceutically acceptable salt thereof, and a compound of
formula: ##STR00159## and/or a pharmaceutically acceptable salt
thereof..Iaddend.
Description
TECHNICAL FIELD OF INVENTION
The present invention relates to compounds useful as intermediates
in the synthesis of pharmaceutically active macrolide
compounds.
BACKGROUND OF THE INVENTION
The invention relates to pharmaceutically active macrolides,
synthesis thereof and intermediates thereto. Halichondrin B is a
potent anticancer agent originally isolated from the marine sponge
Halichondria okadai, and subsequently found in Axinella sp.,
Phakellia carteri, and Lissondendryx sp. A total synthesis of
Halichondrin B was published in 1992 (Aicher, T. D. et al., J. Am.
Chem. Soc. 114: 3162-3164). Halichondrin B has demonstrated in
vitro inhibition of tubulin polymerization, microtubule assembly,
beta.sup.s-tubulin crosslinking, GTP and vinblastine binding to
tubulin, and tubulin-dependent GTP hydrolysis and has shown in
vitro and in vivo anti-cancer properties. Accordingly, there is a
need to develop synthetic methods for preparing analogs of
Halichondrin B useful as anti-cancer agents.
SUMMARY OF THE INVENTION
As described herein, the present invention provides methods for
preparing analogs of Halichondrin B having pharmaceutical activity,
such as anticancer or antimitotic (mitosis-blocking) activity.
These compounds include a compound of formula B-1939:
##STR00002##
These compounds are useful for treating cancer and other
proliferative disorders including, but not limited to, melanoma,
fibrosarcoma, leukemia, colon carcinoma, ovarian carcinoma, breast
carcinoma, osteosarcoma, prostate carcinoma, and lung carcinoma.
The present invention also provides synthetic intermediates useful
for preparing said analogs of Halichondrin B.
DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS OF THE INVENTION
The methods and intermediates of the present invention are useful
for preparing various analogs of Halichondrin B as described in,
e.g. U.S. Pat. No. 6,365,759 and U.S. Pat. No. 6,469,182 the
entirety of which are incorporated herein by reference. These
Halichondrin B analogs are prepared generally by the assembly of
three fragments F-1, F-2, and F-3, as shown by Scheme I below:
##STR00003## 1. Fragment F-1
According to one embodiment, the present invention provides a
compound F-1:
##STR00004## wherein: each of PG.sup.1 and PG.sup.2 is
independently hydrogen or a suitable hydroxyl protecting group;
R.sup.1 is R or OR; R.sup.2 is CHO or --CH.dbd.CH.sub.2; and each R
is independently hydrogen, C.sub.1-4 haloaliphatic, benzyl, or
C.sub.1-4 aliphatic, provided that when R.sup.1 is OMe then
PG.sup.1 and PG.sup.2 do not form an acetonide group.
In certain embodiments, R.sup.1 is OR. In other embodiments,
R.sup.1 is OR wherein R is hydrogen, methyl, or benzyl.
In certain embodiments, PG.sup.1 and PG.sup.2 are hydrogen. In
other embodiments, one of PG.sup.1 and PG.sup.2 is hydrogen.
Suitable hydroxyl protecting groups are well known in the art and
include those described in detail in Protecting Groups in Organic
Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd edition, John
Wiley & Sons, 1999, the entirety of which is incorporated
herein by reference. In certain embodiments, each of PG.sup.1 and
PG.sup.2, taken with the oxygen atom to which it is bound, is
independently selected from esters, ethers, silyl ethers, alkyl
ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of such
esters include formates, acetates, carbonates, and sulfonates.
Specific examples include formate, benzoyl formate, chloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate,
4,4-(ethylenedithio) pentanoate, pivaloate (trimethylacetyl),
crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,
2,4,6-trimethylbenzoate, or carbonates such as methyl,
9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl,
2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and
p-nitrobenzyl. Examples of such silyl ethers include
trimethylsilyl, triethylsilyl, t-butyldimethylsilyl,
t-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl
ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl,
3,4-dimethoxybenzyl, trityl, t-butyl, allyl, and allyloxycarbonyl
ethers or derivatives. Alkoxyalkyl ethers include acetals such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl,
benzyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and
tetrahydropyranyl ethers. Examples of arylalkyl ethers include
benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, o-nitrobenzyl,
p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2-
and 4-picolyl.
In certain embodiments, one or both of the PG.sup.1 and PG.sup.2
moieties of F-1 are silyl ethers or arylalkyl ethers. In yet other
embodiments, one or both of the PG.sup.1 and PG.sup.2 moieties of
F-1 are t-butyldimethylsilyl or benzoyl. In still other
embodiments, both of the PG.sup.1 and PG.sup.2 moieties of F-1 are
t-butyldimethylsilyl.
According to an alternate embodiment, PG.sup.1 and PG.sup.2 are
taken together, with the oxygen atoms to which they are bound, to
form a diol protecting group, such as a cyclic acetal or ketal.
Such groups include methylene, ethylidene, benzylidene,
isopropylidene, cyclohexylidene, and cyclopentylidene, a silylene
derivative such as di-t-butylsilylene and a
1,1,3,3-tetraisopropyldisiloxanylidene derivative, a cyclic
carbonate, and a cyclic boronate. Methods of adding and removing
such hydroxyl protecting groups, and additional protecting groups,
are well-known in the art and available, for example, in P. J.
Kocienski, Protecting Groups, Thieme, 1994, and in T. W. Greene and
P. G. M. Wuts, Protective Groups in Organic Synthesis, 3.sup.rd
edition, John Wiley & Sons, 1999. According to another
embodiment, PG.sup.1 and PG.sup.2 are taken together to form an
acetonide group.
According to one embodiment, R.sup.2 is CHO.
According to another embodiment, R.sup.2 is --CH.dbd.CH.sub.2.
In certain embodiments, the present invention provides a compound
of formula F-1 having the stereochemistry depicted in compound
F-1':
##STR00005## wherein each variable is as defined above and
described in classes and subclasses above and herein.
In certain embodiments, the following compounds F-1a and F-1b are
provided:
##STR00006## wherein "TBS" refers to t-butyldimethylsilyl.
Details of the syntheses of F-1a and F-1b are set forth in the
Examples infra.
2. Fragment F-2
According to another embodiment, the present invention provides a
compound F-2:
##STR00007## wherein: each is independently a single or double
bond, provided that both groups are not simultaneously a double
bond; LG.sup.1 is a suitable leaving group; X is halogen or
--OSO.sub.2(R.sup.y); R.sup.y is C.sub.1-6 aliphatic or a 5-7
membered saturated, partially unsaturated, or fully unsaturated
ring, wherein R.sup.y is optionally substituted with up to 3 groups
selected from halogen, R, NO.sub.2, CN, OR, SR, or N(R).sub.2; each
R is independently hydrogen, C.sub.1-4 haloaliphatic, or C.sub.1-4
aliphatic; and PG.sup.3 is a suitable hydroxyl protecting
group.
As used herein, a suitable leaving group is a chemical moiety that
is readily displaced by a desired incoming chemical moiety.
Suitable leaving groups are well known in the art, e.g., see,
"Advanced Organic Chemistry," Jerry March, 4.sup.th Ed., pp.
351-357, John Wiley and Sons, N.Y. (1992). Such leaving groups
include, but are not limited to, halogen, alkoxy, sulphonyloxy,
optionally substituted alkylsulphonyloxy, optionally substituted
alkenylsulfonyloxy, optionally substituted arylsulfonyloxy, and
diazonium moieties. Examples of suitable leaving groups include
chloro, iodo, bromo, fluoro, methanesulfonyloxy (mesyloxy),
tosyloxy, triflate, nitro-phenylsulfonyloxy (nosyloxy), and
bromo-phenylsulfonyloxy (brosyloxy). In certain embodiments, the
LG.sup.1 moiety of F-2 is sulphonyloxy, optionally substituted
alkylsulphonyloxy, optionally substituted alkenylsulfonyloxy, or
optionally substituted arylsulfonyloxy. In other embodiments, the
LG.sup.1 moiety of F-2 is optionally substituted alkylsulphonyloxy.
In yet other embodiments, the LG.sup.1 moiety of F-2 is mesyloxy or
tosyloxy.
In certain embodiments, the X moiety of F-2 is halogen. In other
embodiments, the X moiety of F-2 is sulphonyloxy, optionally
substituted alkylsulphonyloxy, optionally substituted
alkenylsulfonyloxy, or optionally substituted arylsulfonyloxy. In
still other embodiments, the X moiety of F-2 is triflate.
In certain embodiments, the PG.sup.3 moiety of F-2, taken with the
oxygen atom to which it is bound, is a silyl ether. In other
embodiments, the PG.sup.3 moiety of F-2, taken with the oxygen atom
to which it is bound, is an ester group. According to one aspect of
the present invention, the PG.sup.3 moiety of F-2 is
t-butyldimethylsilyl. According to another aspect of the present
invention, the PG.sup.3 moiety of F-2 is pivaloyl or benzoyl.
In certain embodiments, the present invention provides a compound
of formula F-2 having the stereochemistry depicted in formula
F-2':
##STR00008## wherein each variable is as defined above and
described in classes and subclasses above and herein.
In certain embodiments, a compound F-2a or F-2b is provided:
##STR00009## wherein "MsO" refers to mesylate, "TfO" refers to
triflate, "OPv" refers to pivaloate, "OBz" refers to benzoate, and
"TSO" refers to tosylate.
In other embodiments, the present invention provides a compound of
formula F-2b wherein said compound is crystalline. According to
another embodiment, a compound of formula F-2b is provided wherein
said compound is crystallized from an alkane solvent. In certain
embodiments, crystalline F-2b is provided wherein said compound is
crystallized from pentane or heptane. In other embodiments,
crystalline F-2b is provided wherein said compound is crystallized
at about 0.degree. C.
Compounds of formula F-2 are prepared generally from intermediates
F-2d and F-2e as shown in Scheme A below.
##STR00010##
Accordingly, another aspect of the present invention provides a
compound of formula F-2d:
##STR00011## wherein: R' is --CH.dbd.CH.sub.2 or --C(O)H; Alk is a
C.sub.1-4 straight or branched aliphatic group; and PG.sup.5 is a
suitable hydroxyl protecting group.
Suitable hydroxyl protecting group PG.sup.5 is as described and
defined for the PG.sup.3 moiety of compound F-2, supra. In certain
embodiments, PG.sup.5, taken with the oxygen atom to which it is
bound, is a silyl ether. In other embodiments, PG.sup.5 is
t-butyldimethylsilyl.
According to one embodiment, the Alk moiety of compound F-2d is
methyl.
In certain embodiments, a compound of formula F-2d' is
provided:
##STR00012##
Yet another aspect of the present invention provides a compound of
formula F-2e:
##STR00013## wherein: R'' is OH, OPG.sup.3, or LG.sup.4; LG.sup.4
is a suitable leaving group; and each PG.sup.3 is independently a
suitable hydroxyl protecting group, provided that R'' is other than
OMs when PG.sup.3 is t-butyldiphenylsilyl.
One of ordinary skill in the art would recognize that the R''
moiety of compound F-2e may be transformed from OH to a protected
hydroxyl group, OPG.sup.3, or, alternatively, directly to LG.sup.4.
Such transformations are known to one skilled in the art and
include, among others, those described herein. In certain
embodiments, R'' is OH or LG.sup.4. The LG.sup.4 leaving group of
formula F-2e is as described and defined for the LG.sup.1 moiety of
compound F-2, supra. In certain embodiments, LG.sup.4 is tosyloxy
or mesyloxy.
The PG.sup.3 moiety of compound F-2e is as defined and described
for the PG.sup.3 moiety of compound F-2, supra. In certain
embodiments, PG.sup.3, taken with the oxygen atom to which it is
bound, is a silyl ether. In other embodiments, PG.sup.3 is
t-butyldiphenylsilyl.
Still another aspect of the present invention provides a compound
F-2f:
##STR00014## wherein Alk, PG.sup.3 and PG.sup.5 are as defined
generally and in classes and subclasses described above and herein.
Compounds of formula F-2f are used to prepare compounds of formula
F-2 by methods described herein and those known in the art.
Details of the synthesis of F-2a are set forth in the Examples
infra.
Alternatively, compounds of formula F-2 are prepared from D-quinic
acid as shown by Scheme II below. Details of the preparation of
compounds of formula F-2 are set forth in the Examples infra.
##STR00015## ##STR00016## ##STR00017##
Yet another method for preparing compounds of formula F-2 from
D-quinic acid provides an alternative route from intermediate 12 to
intermediate 17 as shown in Scheme III below.
##STR00018##
Scheme III above shows an alternate method for preparing
intermediate 17 from intermediate 12 via Eschenmoser-Tanabe
Fragmentation, wherein each .[.Rx.]. .Iadd.R.sup.x.Iaddend. is
independently OPG.sup.x or CN wherein PG.sup.x is a suitable
hydroxyl protecting group as described herein. Intermediate 17 is
then used to prepare compounds of formula F-2 according to Scheme
II above.
Still another method for preparing compounds of formula F-2 from
D-quinic acid provides an alternative route from intermediate 9 to
intermediate 17 as shown in Scheme IV below.
##STR00019## ##STR00020## wherein PG.sup.y is a suitable carboxyl
protecting group, as described herein, and each PG.sup.x is
independently a suitable hydroxyl protecting group as described
herein.
Yet another method for preparing intermediates useful for preparing
compounds of formula F-2 from D-quinic acid is shown in Scheme V
below.
##STR00021##
In Scheme V above, intermediate 7 (from Scheme II), wherein R is a
methyl ester, is used to prepare ER-817664 as a crystalline
intermediate. As depicted in Scheme V above, each PG.sup.5 and
PG.sup.6 is independently a suitable hydroxyl protecting group. In
certain embodiments, PG.sup.5 and PG.sup.6 are taken together to
form a cyclic diol protecting group. In other embodiments, PG.sup.5
and PG.sup.6 are taken together to form a cyclohexylidene
protecting group. As depicted in Scheme V above, LG.sup.5 is a
suitable leaving group. Such suitable leaving groups are well known
in the art and include those described herein. In certain
embodiments, LG.sup.5 is mesyloxy or tosyloxy.
According to another embodiment, the present invention provides a
compound of formula A:
##STR00022## wherein: designates a single or double bond; n is 1,
2, or 3; each of PG.sup.5 and PG.sup.6 is independently a suitable
hydroxyl protecting group; W is CH-A or C(O); A is a C.sub.1-6
aliphatic group, wherein A is optionally substituted with one or
more Q.sup.1 groups; each Q.sup.1 is independently selected from
cyano, halo, azido, oxo, OR, SR, SO.sub.2R, OSO.sub.2R, N(R).sub.2,
NR(CO)R, NR(CO) (CO)R, NR(CO)N(R).sub.2, NR(CO)OR, (CO)OR, O(CO)R,
(CO)N(R).sub.2, O(CO)N(R).sub.2, or OPG.sup.1, wherein PG.sup.1 is
a suitable hydroxyl protecting group, and wherein: two Q.sup.1 on A
are optionally taken together to form a 3-8 membered saturated,
partially unsaturated, or aryl ring having 0-4 heteroatoms
independently selected from nitrogen, oxygen, or sulfur; and each R
is independently selected from hydrogen or an optionally
substituted group selected from C.sub.1-6 aliphatic, a 5-10
membered saturated, partially unsaturated or aryl carbocyclic ring,
or a 4-10 membered saturated, partially unsaturated or aryl ring
having 0-4 heteroatoms independently selected from nitrogen,
oxygen, or sulfur, wherein: two R groups on the same nitrogen atom
are optionally taken together with said nitrogen atom to form a 3-8
membered saturated, partially unsaturated, or aryl ring having 1-4
heteroatoms independently selected from nitrogen, oxygen, or
sulfur.
In certain embodiments, the present invention provides a compound
of formula A having the stereochemistry as depicted in formula
A':
##STR00023## wherein each variable is as defined above and
described in classes and subclasses above and herein.
In certain embodiments, the present invention provides a compound
of formula A' wherein W is C(O) and said compound is of formula
A'-1:
##STR00024## wherein each variable is as defined above and
described in classes and subclasses above and herein.
As defined generally above, the A group of formulae A and A' is a
C.sub.1-6 aliphatic group, wherein A is optionally substituted with
Q.sup.1. In certain embodiments, the A group of formulae A and A'
is a C.sub.2-5 aliphatic group, wherein A is substituted with one
or more Q.sup.1 groups.
As defined generally above, each Q.sup.1 group of formulae A and A'
is independently selected from cyano, halo, azido, oxo, OR, SR,
SO.sub.2R, OSO.sub.2R, N(R).sub.2, NR(CO)R, NR(CO) (CO)R,
NR(CO)N(R).sub.2, NR(CO)OR, (CO)OR, O(CO)R, (CO)N(R).sub.2,
O(CO)N(R).sub.2, or OPG.sup.1, wherein PG.sup.1 is a suitable
hydroxyl protecting group. In certain embodiments, each Q.sup.1
group of formulae A and A' is independently selected from cyano,
halo, azido, oxo, N(R).sub.2, OR, SR, SO.sub.2R, or OSO.sub.2R. In
other embodiments, each Q.sup.1 group of formulae A and A' is
independently selected from cyano, halo, azido, oxo, OR, SR,
SO.sub.2R, OSO.sub.2R, N(R).sub.2, NR(CO)R, NR(CO)R, and O(CO)N
(R).sub.2. In still other embodiments, exemplary Q.sup.1 groups
include NH(CO) (CO)-(heterocyclic radical or heteroaryl),
OSO.sub.2-(aryl or substituted aryl), O(CO)NH-(aryl or substituted
aryl), aminoalkyl, hydroxyalkyl, NH(CO) (CO)-(aryl or substituted
aryl), NH(CO) (alkyl) (heteroaryl or heterocyclic radical),
O(substituted or unsubstituted alkyl) (substituted or unsubstituted
aryl), and NH(CO) (alkyl) (aryl or substituted aryl).
In certain embodiments, the A group of formulae A and A' has one of
the following characteristics: (1) A has at least one substituent
selected from hydroxyl, amino, azido, halo, and oxo; (2) A is a
C.sub.1-6 alkyl group having at least one substituent selected from
hydroxyl, amino, and azido; (3) A has at least two substituents
independently selected from hydroxyl, amino, and azido; (4) A has
at least two substituents independently selected from hydroxyl and
amino; (5) A has at least one hydroxyl substituent and at least one
amino substituent; (6) A has at least two hydroxyl substituents;
(7) A is a C.sub.2-4 aliphatic group that is substituted; (8) A is
a C.sub.3 aliphatic group that is substituted; (9) A has an
(S)-hydroxyl alpha to the carbon atom linking A to the ring
containing G or an (R)-hydroxyl; and (10) A is a C.sub.1-6
saturated aliphatic group having at least one substituent selected
from hydroxyl and cyano.
The term "(S)-hydroxyl" means that the configuration of the carbon
atom having the hydroxyl group is (S). Embodiments of the invention
also include compounds wherein A is substituted at least once on
each carbon atom: (1) alpha and gamma, (2) beta and gamma, or (3)
alpha and beta to the carbon atom to which A is attached. Each of
the alpha, beta, and gamma carbon atoms are independently in the
(R) or (S) configuration. In certain embodiments, the invention
provides said compound wherein A is substituted at least once on
each carbon atom alpha and beta to the carbon atom to which A is
attached.
Exemplary A groups of formulae A and A' include
2,3-dihydroxypropyl, 2-hydroxyethyl, 3-hydroxy-4-perfluorobutyl,
2,4,5-trihydroxypentyl, 3-amino-2-hydroxypropyl,
1,2-dihydroxyethyl, .[.2,3-dihyroxy-4-perflurobutyl.].
.Iadd.2,3-dihydroxy-4-perfluorobutyl.Iaddend.,
3-cyano-2-hydroxypropyl, 2-amino-1-hydroxy ethyl,
3-azido-2-hydroxypropyl, 3,3-difluoro-2,4-dihydroxybutyl,
2,4-dihydroxybutyl, 2-hydroxy-2-(p-fluorophenyl)-ethyl,
--CH.sub.2(CO) (substituted or unsubstituted aryl), --CH.sub.2(CO)
(alkyl or substituted alkyl, such as haloalkyl or hydroxyalkyl) and
3,3-difluoro-2-hydroxypent-4-enyl.
In certain embodiments, the A group of either of formulae A and A'
is 3-amino-2-hydroxypropyl.
According to one aspect, the present invention provides a compound
of either of formulae A and A', wherein Q.sup.1 is OPG.sup.1,
wherein PG.sup.1 is a suitable hydroxyl protecting group. Suitable
hydroxyl protecting groups are well known in the art and include
those described in detail in Protecting Groups in Organic
Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd edition, John
Wiley & Sons, 1999, the entirety of which is incorporated
herein by reference. In certain embodiments, the PG.sup.1 moiety of
either of formulae A and A', taken with the oxygen atom to which it
is bound, is selected from esters, ethers, silyl ethers, alkyl
ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of such
esters include formates, acetates, carbonates, and sulfonates.
Specific examples include formate, benzoyl formate, chloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate,
4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetyl),
crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,
2,4,6-trimethylbenzoate, carbonates such as methyl,
9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl,
2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and
p-nitrobenzyl. Examples of such silyl ethers include
trimethylsilyl, triethylsilyl, t-butyldimethylsilyl,
t-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl
ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl,
3,4-dimethoxybenzyl, trityl, t-butyl, allyl, and allyloxycarbonyl
ethers or derivatives. Alkoxyalkyl ethers include acetals such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl,
benzyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and
tetrahydropyranyl ethers. Examples of arylalkyl ethers include
benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, o-nitrobenzyl,
p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2-
and 4-picolyl.
In certain embodiments, the PG.sup.1 moiety of either of formulae A
and A', taken with the oxygen atom to which it is bound, is a silyl
ether or arylalkyl ether. In yet other embodiments, the PG.sup.1
moiety of either of formulae A and A' is t-butyldimethylsilyl or
benzoyl. In still other embodiments, the PG.sup.1 moiety of either
of formulae A and A' is t-butyldimethylsilyl ("TBS").
As defined generally above, two Q.sup.1 on A are optionally taken
together to form a 3-8 membered saturated, partially unsaturated,
or aryl ring having 0-4 heteroatoms independently selected from
nitrogen, oxygen, or sulfur. In certain embodiments, two Q.sup.1 on
A are taken together to form an epoxide ring.
In certain embodiments, the PG.sup.5 and PG.sup.6 groups of formula
A and A' are independently selected from those suitable protecting
groups described above for the PG.sup.1 group of formula A and A'.
In other embodiments, the PG.sup.5 and PG.sup.6 groups of formula A
and A' are taken together to form a cyclic diol protecting group.
Such diol protecting groups are well known in the art and include
those described by Greene and include cyclohexylidene and
benzylidene diol protecting groups.
In certain embodiments, the present invention provides a method for
preparing compounds of formula F-2 according to Schemes V-a, V-b,
and V-c below:
##STR00025## ##STR00026##
In other embodiments, the present invention provides crystalline
ER-817664.
##STR00027## ##STR00028##
##STR00029##
Using ER-817664 as a crystalline intermediate, Scheme VI-a shows a
general method for using this compound in the preparation of
intermediates useful for preparing compounds of formula F-2.
##STR00030##
The triol intermediate depicted in Scheme VI-a above is used in an
alternate method for preparing intermediates useful for preparing
compounds of formula F-2, as shown in Scheme VI-b below.
##STR00031##
In Scheme VI-b, shown above, the triol intermediate is treated with
periodate to form the aldehyde. This compound is homologated with
methyl Wittig reagent, and the resulting olefin reduced, to form
the ester compound. The remaining free hydroxyl group is treated
with N-iodosuccinimide to form the iodo intermediate and the ester
reduced with sodium borohydride to form the hydroxyl compound
depicted above. One of ordinary skill in the art will recognize
that the resulting iodo compound corresponds to compound 21
depicted in Scheme II, supra, wherein compound 21 has a protecting
group at the hydroxyl position. The final treatment with zinc
affords the lactone depicted above. One of ordinary skill in the
art will recognize that the resulting lactone compound corresponds
to compound 22 depicted in Scheme II, supra, wherein compound 22
has a protecting group at the hydroxyl position.
Yet another alternate method for preparing intermediates useful for
preparing compounds of formula F-2 from D-quinic acid provides an
alternative route from intermediate 2, of Scheme II as shown in
Scheme VII below.
##STR00032##
In Scheme VII above, intermediate 2 (from Scheme II) is used to
prepare ER-812829 in a stereoselective manner. It will be
appreciated that other protecting groups are useful for protecting
the diol of ER-812829. Such groups are known to one of ordinary
skill in the art and include cyclohexylidene and benzylidene diol
protecting groups. First, at step (a), the hydroxyl group of
ER-811510 is treated with 2-bromo acetylchloride to form ER-812771.
The bromo intermediate is treated with triphenylphosphine to form a
Wittig reagent in situ in a manner substantially similar to that
described by Murphy, et al, Tetrahedron Letters, 40, (1999)
3455-3456. This Wittig reagent then forms the lactone ER-812772. At
step (d), stereoselective hydrogenation of the double bond affords
ER-812829.
The present invention also provides a method for preparing
intermediates useful for preparing compounds of formula F-2 from
D-quinic acid, from intermediate ER-812829 depicted in Scheme VII
above, as shown in Scheme VII-a below.
##STR00033## ##STR00034## 3. Fragment F-3
According to yet another embodiment, the present invention provides
a compound F-3:
##STR00035## wherein: each PG.sup.4 is an independently selected
suitable hydroxyl protecting group; R.sup.3 is CHO or C(O)OR.sup.4;
R.sup.4 is a suitable carboxyl protecting group; and LG.sup.2 is a
suitable leaving group.
Suitable carboxylate protecting groups are well known in the art
and are described in detail in Protecting Groups in Organic
Synthesis, T. W. Greene and P. G. M. Wuts, 3.sup.rd edition, John
Wiley & Sons, 1999. In certain embodiments, the R.sup.4 group
of F-3 is an optionally substituted C.sub.1-6 aliphatic group or an
optionally substituted aryl group. Examples of suitable R.sup.4
groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl,
benzyl, and phenyl wherein each group is optionally
substituted.
As described above, suitable leaving groups are well known in the
art, e.g., see "Advanced Organic Chemistry," Jerry March, 4.sup.th
Ed., pp. 351-357, John Wiley and Sons, N.Y. (1992). Such leaving
groups include, but are not limited to, halogen, alkoxy,
sulphonyloxy, optionally substituted alkylsulphonyloxy, optionally
substituted alkenylsulfonyloxy, optionally substituted
arylsulfonyloxy, silyl, and diazonium moieties. Examples of
suitable leaving groups include chloro, iodo, bromo, fluoro,
methanesulfonyloxy (mesyloxy), tosyloxy, triflate,
nitro-phenylsulfonyloxy (nosyloxy), and bromo-phenylsulfonyloxy
(brosyloxy). In certain embodiments, the LG.sup.2 moiety of F-3 is
iodo.
According to an alternate embodiment, the suitable leaving group
may be generated in situ within the reaction medium. For example,
LG.sup.2 in a compound of formula F-3 may be generated in situ from
a precursor of that compound of formula F-3 wherein said precursor
contains a group readily replaced by LG.sup.2 in situ. In a
specific illustration of such a replacement, said precursor of a
compound of formula F-3 contains a group (for example, a
trimethylsilyl group) which is replaced in situ by LG.sup.2, such
as an iodo group. The source of the iodo group may be, e.g.,
N-iodosuccinimide. Such an in situ generation of a suitable leaving
group is well known in the art, e.g., see Id.
As described above, suitable hydroxyl protecting groups are well
known in the art and include those described in detail in
Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M.
Wuts, 3.sup.rd edition, John Wiley & Sons, 1999 the entirety of
which is incorporated herein by reference. In certain embodiments,
each PG.sup.4, taken with the oxygen atom to which it is bound, is
independently selected from esters, ethers, silyl ethers, alkyl
ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of such
esters include formates, acetates, carbonates, and sulfonates.
Specific examples include formate, benzoyl formate, chloroacetate,
trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,
p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate,
4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetyl),
crotonate, 4-methoxy-crotonate, benzoate, p-benzylbenzoate,
2,4,6-trimethylbenzoate, carbonates such as methyl,
9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl,
2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and
p-nitrobenzyl. Examples of such silyl ethers include
trimethylsilyl, triethylsilyl, t-butyldimethylsilyl,
t-butyldiphenylsilyl, triisopropylsilyl, and other trialkylsilyl
ethers. Alkyl ethers include methyl, benzyl, p-methoxybenzyl,
3,4-dimethoxybenzyl, trityl, t-butyl, allyl, and allyloxycarbonyl
ethers or derivatives. Alkoxyalkyl ethers include acetals such as
methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl,
benzyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and
tetrahydropyranyl ethers. Examples of arylalkyl ethers include
benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, o-nitrobenzyl,
p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2-
and 4-picolyl.
In certain embodiments, one, two, or three of the PG.sup.4 moieties
of F-3, taken with the oxygen atom(s) to which they are bound, are
silyl ethers or arylalkyl ethers. In yet other embodiments, one,
two, or three of the PG.sup.4 moieties of F-3 are
t-butyldimethylsilyl or benzyl. In still other embodiments, all
three of the PG.sup.4 moieties of F-3 are t-butyldimethylsilyl.
According to another embodiment, a compound of formula F-3 is
provided wherein said compound has the stereochemistry as depicted
in formula F-3':
##STR00036## wherein each variable is as defined above and
described in classes and subclasses above and herein.
In certain embodiments, a compound F-3a is provided:
##STR00037## wherein "TBS" refers to t-butyldimethylsilyl.
Details of the synthesis of F-3a are set forth in the Examples
infra.
4. Assembly of F-1, F-2, and F-3 to Prepare Compound I
Coupling of the fragments F-1 and F-2 is accomplished, in general,
as set forth in Scheme VIII below.
##STR00038##
Scheme VIII above shows a general method for preparing intermediate
F-5a from fragments F-1 and F-2. First, fragments F-1 and F-2 are
coupled using methods substantially similar to that described by
Kishi, et al., Org Lett 4:25 p 4431 (2002) to afford intermediate
F-4. This coupling is performed in the presence of the chiral
oxazole (ER-807363) or, alternatively, in the absence of ER-807363.
However, the coupling reaction of F-1 and F-2 proceeds with higher
selectivity when performed in the presence of ER-807363.
Intramolecular Williamson ether formation of F-4, by treating F-4
with potassium hexamethyldisilazide, then furnishes tetrahydropyran
F-5 as a mixture of stereoisomers. The stereoisomers are then
separated to afford F-5a. The details of these steps are set forth
in the Examples infra.
According to another embodiment, the present invention provides a
compound F-4:
##STR00039## wherein PG.sup.1, PG.sup.2, PG.sup.3, LG.sup.1, and
R.sup.1 are as defined in general and in subclasses above and
herein. .Iadd.PG.sup.3 is hydrogen or a suitable hydroxyl
protecting group..Iaddend.
In certain embodiments, the present invention provides a compound
of formula F-4 wherein said compound has the stereochemistry
depicted in formula F-4':
##STR00040## wherein PG.sup.1, PG.sup.2, PG.sup.3, LG.sup.1, and
R.sup.1 are as defined in general and in subclasses above and
herein.
The present invention also provides a compound F-4a:
##STR00041## wherein "MsO" refers to mesylate, "TBS" refers to
t-butyldimethylsilyl, and "OPv" refers to pivaloate.
Details of the synthesis of F-4a are set forth in the Examples
infra.
According to yet another embodiment, the present invention provides
a compound F-5:
##STR00042## wherein each PG.sup.1, PG.sup.2, PG.sup.3, and R.sup.1
is as defined in general and in subclasses above and herein.
In certain embodiments, the present invention provides a compound
of formula F-5 having the stereochemistry as depicted in formula
F-5' or F-5a:
##STR00043## wherein each PG.sup.1, PG.sup.2, PG.sup.3, and R.sup.1
is as defined in general and in subclasses above and herein.
The PG.sup.3 group of intermediate F-5a is removed and the
resulting hydroxyl compound F-6 is then coupled with a compound
F-3', wherein R.sup.3 is CHO, to form F-7 as depicted in Scheme IX
below.
##STR00044##
Scheme IX above shows a general method for preparing an
intermediate-F-9 from F-3'and F-6. First, the sulfone intermediate
F-6 is treated with n-butyl lithium then with the aldehyde F-3'.
The resulting diol intermediate F-7 is then oxidized with
Dess-Martin reagent to form the ketone-aldehyde intermediate F-8
which is then treated with SmI.sub.2 to afford intermediate F-9.
The details of these steps are set forth in the Examples infra.
##STR00045##
Scheme X above sets forth a general method for preparing the
Halichondrin B analogs of the present invention from F-9a (LG.sup.2
is iodo). First, an intramolecular coupling is achieved, by
conditions substantially similar to those described at Scheme V
above, to form hydroxyl compound F-10. In an alternate method, the
intramolecular coupling is performed in the presence of the chiral
oxazole ligand, described herein. The addition of the chiral
oxazole ligand imparts a higher yield and greater efficiency for
the reaction. The details of this reaction are set forth in the
Examples below. Compound F-10 is then oxidized to form F-11. The
hydroxyl protecting groups of F-11 are removed by appropriate means
to afford F-12. One of ordinary skill in the art would recognize
that the methods appropriate to achieve removal of the protecting
groups of compound F-11 depend upon the actual protecting groups
used and include those described by Greene. For example, when each
of the hydroxyl protecting groups of F-11 is a TBS group, such
removal may be achieved by treatment with optionally buffered
tetrabutylammonium fluoride. The details of these steps are set
forth in the Examples infra.
Intermediate F-12 is useful for preparing various analogs of
Halichondrin B as described in, e.g. U.S. Pat. Nos. 6,365,759 and
6,469,182 the entirety of which are incorporated herein by
reference.
EXAMPLES
Using the preparation of Halichondrin B analog B-1939 to exemplify,
the following Examples describe the synthesis of Halichondrin B
analogs using the methods and compounds of the present
invention.
##STR00046##
One of ordinary skill in the art would recognize that many analogs
of Halichondrin B are prepared by the methods and from the
compounds of the present invention including, but not limited to,
those analogs of Halichondrin B described in U.S. Pat. Nos.
6,214,865 and 6,365,759, the entirety of which are herein
incorporated by reference. Accordingly, it will be appreciated that
the synthetic methods described below, by way of example, do not
limit the scope of the invention which is defined by the appended
claims.
Example 1
Preparation of F-1a
##STR00047##
In an appropriately sized vessel, D-glucurono-6,3-lactone (1 wt., 1
eq.) was combined with ACN (3 vol.) and acetone (9 vol.). Catalytic
conc. sulfuric acid was added and the system held at reflux for 3
hours. The system was checked for dissolution of
D-glucurono-6,3-lactone. The reaction was cooled to 25.degree. C.
and stirred for 15 hours. Solid sodium bicarbonate (0.5 wts) was
added and the reaction stirred for 3 additional hours. Solids were
removed by filtration and the organics were partially concentrated
and azeotroped with additional ACN (2 wts). ER-806045 was taken
into the next reaction without isolation.
##STR00048##
Crude ER-806045 (1 wt, 1 eq.) was dissolved in ACN (6.5 vol.) at
-20.degree. C. Pyridine (1.5 vol., 4.0 eq.) was added and
SO.sub.2Cl.sub.2 (0.38 vol., 1.02 eq.) was added slowly, while
keeping the internal temperature below 5.degree. C. The reaction
was quenched by inverse addition into cool water (28 vol.) with an
ACN rinse (0.5 vol.), keeping the internal temperature below
10.degree. C. The white solid, ER-806410 (0.87 wt., 79% of
theoretical) was isolated by filtration with a heptane rinse (2
vol.) and drying.
##STR00049##
An appropriately sized vessel was charged with ER-806410 (1 wt, 1
eq.) and THF (10 vol.) and then cooled to 10.degree. C. Wet
palladium on carbon (5%, 0.5 wts) was added and the heterogeneous
solution stirred for ten minutes. The reaction was buffered with
pyridine (0.44 wts, 1.3 eq.) and placed under a hydrogen atmosphere
for 3 hours. The reaction was filtered and the solids rinsed with
water (2 vol.) and EtOAc (10 vol.). The resulting solution was
acidified with 1N HCl (2.1 vol.), mixed well and the resulting
layers were separated. The organic layer was sequentially washed
with aqueous sodium bicarbonate (5 vol.) and water (5 vol.). The
organics were concentrated under reduced pressure and the resulting
product recrystallized from IPA (3.4 vol.) and further cropped by
the addition of heptane (3.4 vol.) at 15.degree. C. ER-806047 was
isolated as a white solid (67% yield).
##STR00050##
An appropriately sized vessel was charged with ER-806047 (1 wt, 1
eq.) and toluene (8 vol.) and then cooled to -40.degree. C. A 17 wt
% solution of DIBAL in toluene (4.6 wts, 1.1 eq.) was added,
keeping the internal temperature below -35.degree. C. After
assaying the reaction, excess reagent was quenched by the addition
of acetone (0.15 wts, 0.5 eq.), keeping the temperature below
10.degree. C. The reaction was diluted with EtOAc (7 vol.) and 15%
aqueous citric acid (8 wts) below 10.degree. C.; and stirred at
20.degree. C. until a clear solution was obtained. The layers were
separated and the aqueous layer back extracted twice with EtOAc
(2.times.10 vol.). The combined organics were washed sequentially
with aqueous sodium bicarbonate (5 vol.) and brine (5 vol.) and
then dried with magnesium sulfate (0.2 wts). After filtration, the
organic layers were partially concentrated at reduced pressure, and
azeotroped with toluene (4 vol.). The products were stored as a THF
solution for use in the next reaction.
##STR00051##
An appropriately sized vessel was charged with a 20 wt % ether
solution of TMSCH.sub.2MgCl (2.04 wts, 3.0 eq.) and chilled below
5.degree. C. A THF (7 vol.) solution of ER-806048 (1 wt, 1 eq.) was
added to the reaction vessel keeping the internal temperature below
15.degree. C. The reaction was warmed to 35.degree. C. for 1.5
hours. The reaction was cooled, diluted with toluene (7 vol.) and
quenched with AcOH (3 vol.) below 20.degree. C. The reaction was
further diluted with 10% aqueous ammonium chloride (6 wts), mixed
well, and the layers were separated. The organic layer was washed
sequentially with aqueous sodium bicarbonate (5 vol.) and brine (5
vol.). After drying over magnesium sulfate (0.2 wts) and
filtration, the solution was concentrated under reduced pressure
and ER-807114 isolated as a concentrated solution in toluene (90%
yield).
##STR00052##
An appropriately sized vessel was charged sequentially with
ER-807114 (1 wt, 1 eq.) and THF (20 vol.), and the solution was
cooled below 5.degree. C. A 15 wt % solution of KHMDS in toluene
(9.16 wts, 2.0 eq.) was added. The reaction was quenched with 10%
aqueous ammonium chloride (5 vol.). The layers were separated and
the organic layer washed sequentially with ammonium chloride (5
vol.), 2N HCl (8.5 vol.), aqueous sodium bicarbonate (5 vol.), and
water (5 vol.). The organics were transferred to concentration
vessels using EtOAc, and concentrated to a viscous oil (90% yield).
The material was recrystallized from toluene (4 vol.) and heptane
(4 vol.) at 35.degree. C. with additional cropping at lower
temperatures and heptane (2.times.4 vol.) at 15 and 10.degree. C.
(94% yield).
##STR00053##
An appropriately sized vessel was charged with KOtBu (0.67 wts, 1.2
eq.) and THF (7.7 vol.), and cooled to an internal temperature of
-20.degree. C. A solution of ER-806049 (1 wt, 1 eq.) in THF (2.3
vol.) was added keeping the internal temperature below -7.degree.
C. Neat BnBr was added, maintaining -7.degree. C. as the maximum
temperature. The reaction was stirred at -20.degree. C. for 2 hours
and 10 hours at 10.degree. C. The reaction was quenched with 10%
aqueous NH.sub.4Cl (4 wts), diluted with toluene (4 vol.), and
mixed well. The layers were separated and the organic layer washed
with 10% brine (4 wts) and dried over MgSO.sub.4 (0.15 wts).
ER-806050 was isolated as a tBuOH solution (2.5 vol.) after
concentration at reduced pressure (95% yield).
##STR00054##
An appropriately sized vessel was charged sequentially with
K.sub.3Fe(CN).sub.6 (3.5 wt., 3.4 eq.), K.sub.2CO.sub.3 (1.5 wt.,
3.4 eq.), (DHQ).sub.2AQN (0.0134 wt., 0.005 eq.), water (18 vol.),
t-BuOH (13 vol.), and ER-806050 in tBuOH (1 wt, 1 eq. in 5 vol.).
The heterogeneous mixture was cooled to an internal temperature of
0.degree. C., and K.sub.2OsO.sub.4.2H.sub.2O (0.0029 wt., 0.22 mole
%) was added. After 36 hours at 0.degree. C., the reaction was
quenched with Na.sub.2S.sub.2O.sub.3 (3.5 eq., 1.7 wt.) and the
flask allowed to warm to ambient temperature overnight. After 15
hours, the mixture was transferred to a workup vessel and diluted
with toluene (15 vol.) and water (4 vol.). The biphasic mixture was
vigorously stirred and separated. The organic layer was washed with
brine (10 vol.), and concentrated and solvent exchanged to afford a
crude mixture of diols ER-806051 and ER-806052 as a 10% toluene
solution (92% yield).
##STR00055##
The toluene solution (10.1 wt %, 9.9 wts) of ER-806051/52 (1 wt, 1
eq.) was further diluted with additional toluene (3 wts).
N-Methylmorpholine (0.94 wts, 3.0 eq.) and DMAP (0.075 wts, 0.2
eq.) were added to the toluene solution and the resulting mixture
was cooled below 15.degree. C. Benzoyl chloride was added keeping
the internal temperature below 25.degree. C. The reaction was then
stirred for 12 hours at 75.degree. C. The reaction was cooled to
15.degree. C. and the temperature kept below 25.degree. C. during
the 1N HCl (5 vol.) quench. Layers were mixed well and separated.
The organic layer was sequentially washed with brine (3 wts),
aqueous sodium bicarbonate (3 wts), and brine (3 wts). The organic
layer was dried (MgSO.sub.4, 0.25 wts), treated with activated
carbon (0.1 wts), and filtered (Celite.RTM., 0.3 wts) with a
toluene (1 wt). The products were partially concentrated under
reduced pressure, azeotroped with toluene (3 wts). Bisbenzoate
ER-806053/54 was isolated in 95% yield as a toluene solution (5
vol.).
##STR00056##
Under an inert atmosphere, a 20 wt % solution of TiCl.sub.4 (6.42
wts, 3.6 eq.) in toluene was cooled to 15.degree. C. Keeping the
internal temperature below 30.degree. C., Ti(OiPr).sub.4 (0.64 wts,
1.2 eq.) was added, and the resulting solution stirred for 15
minutes. AllylTMS (1.03 wts, 4.8 eq.) was premixed with
ER-806053/54 (1 wt, 1 eq.), available as a 22 wt % solution in
toluene from the previous step (4.55 wts, 1 eq.), and added to the
freshly generated Ti(OiPr)Cl.sub.3. The internal temperature during
the addition was kept below 30.degree. C. The reaction was stirred
between 20-30.degree. C. for 2 hours. The reaction was cooled to
-5.degree. C. and quenched with 1N HCl (6 vol.), keeping the
internal temperature below 30.degree. C. After mixing well, the
layers were separated and the organic layer sequentially washed
with 1N HCl (3 vol.) and brine (2.times.3 vol.). The organic layer
was stirred with MgSO.sub.4 (0.3 wts) and activated carbon (0.15
wts) and filtered through a Celite.RTM. plug (0.2 wts), rinsing
with toluene (1 vol.). The product, as a 3:1 mixture at C-34 was
isolated after concentration in 83% yield. Recrystallization from
IPA/n-heptane afforded ER-806055 with .[.99.5%.]. .Iadd.>99.5%
.Iaddend.d.e. at C-34 (71% yield).
##STR00057##
At room temperature, an appropriately sized vessel was charged with
alcohol ER-806055 (1 wt., 1.0 eq.), toluene (7 vol.), DMSO (0.31
wt., 2.0 eq.) and Et.sub.3N (0.78 wt., 4.0 eq.). The resulting
solution was cooled to -19.degree. C. TCAA (0.84 wt., 1.4 eq.) was
added drop-wise, keeping the internal temperature below -10.degree.
C. The reaction was stirred for an additional 10 minutes. The
reaction was diluted with IPA (0.5 vol.) and quenched with 1N HCl
(5 vol.), keeping the internal temperature below 10.degree. C. The
layers were separated and the organic layer sequentially washed
with aqueous NaHCO.sub.3 (5 wt.) and water (3 vol.). The organic
layer was partially concentrated at reduced pressure (100% crude
yield) and further azeotroped with additional toluene (4 vol.). The
resulting ketone (ER-806058) was dissolved in a final 4 vol. of
toluene, checked for water content and used as is in the next
reaction.
##STR00058##
A solution of ER-107446 (1 wt, 1.5 eq.) in THF (2.7 vol.) was
cooled to 10.degree. C. and treated with 25.5 wt % LHMDS in THF
(5.2 wt., 1.4 eq.), keeping the internal temperature below
15.degree. C. In a second vessel, the toluene solution of crude
ER-806058 (21.9 wt %, 5.4 vol.) was cooled to 10.degree. C. The
contents of vessel one were transferred into the substrate
containing solution, keeping the internal temperature below
20.degree. C. The reaction was stirred for 30 minutes then quenched
by adding 1M HCl (6.5 vol.), keeping the internal temperature below
20.degree. C. The layers were separated and the organic layer
washed four times with 1:1 MeOH/water (4.times.5 vol.) and then
with aqueous bicarbonate (5 vol.), and brine solutions (2.times.5
vol.). The product was dried over MgSO.sub.4 (0.52 wt.), filtered
(rinsing with 0.7 vol. of toluene), and concentrated to a heavy oil
at reduced pressure.
##STR00059##
ER-806059 was dissolved in 1:1 toluene/CH.sub.3CN (5 vol.) at room
temperature. Filtered TMSI (1.23 wt., 4 eq.) was added, keeping the
initial temperature below 40.degree. C. The reaction was heated to
60.degree. C. for 2 hours. The reaction was cooled to -15.degree.
and quenched with 25% aqueous ammonium hydroxide below 30.degree.
C. The reaction contents were stirred overnight and the layers
separated. The organic layer was charged with additional toluene (5
vol.) and water (2 vol.). The layers were mixed well and separated.
The organic layer was then washed sequentially with 10% aqueous
sodium sulfite (5 vol.), 1N HCl (5 vol.), 5% aqueous sodium
bicarbonate (5 vol.), and brine (5 vol.). The organic layer was
dried over MgSO.sub.4 (0.2 wt.), filtered, partially concentrated
and used in the next reaction as a 50% solution in toluene.
##STR00060##
In an appropriately sized vessel, NaBH(OAc).sub.3 (1.19 wt, 3.15
eq.), Bu.sub.4NCl (1.04. wt, 2.1 eq.), DME (8.2 vol.), and toluene
(4 vol.) at 65.degree. C., were combined and stirred at room
temperature. The mixture was heated to 75.degree. C. for one hour.
ER-806060 (1 wt., 1 eq.) as a 50% wt solution in toluene was added
at 75.degree. C. and rinsed in with additional toluene (0.3 vol.).
The reaction temperature was raised to 85.degree. C. and the
reaction stirred for 2-4 hours. The reaction was cooled to
<10.degree. C. and quenched with water (3.2 vol.) keeping the
internal temperature below 20.degree. C. The layers were mixed well
and separated. The organic layer was sequentially washed with
aqueous sodium bicarbonate (2.times.5 vol.) and water (2.times.5
vol.). The organic layer was concentrated and solvent exchanged to
afford a 40 wt % solution of ER-806061 in MeOH.
##STR00061##
A 40 wt % solution of ER-806061 (1 wt., 1.0 eq.) in MeOH was
dissolved in additional methanol (1.6 vol.). Potassium carbonate
(0.24 wts, 1.0 eq.) was added and the reaction temperature was
raised to 50.degree. C. for one hour. The reaction was cooled to
15.degree. C., and quenched with 1N HCl (3.5 vol., 2 eq.) with the
internal temperature below 30.degree. C. The reaction was diluted
with water (3.9 vol.) and toluene (3 vol.). The layers were
separated and the aqueous layer back extracted with toluene (1.5
vol.). The aqueous phase was charged with sodium bicarbonate (0.3
wts) and sodium chloride (0.6 wts), and back extracted with nBuOH
(3 vol.). The three organic phases were combined and concentrated
to dryness to afford crude triol ER-806064 and inorganic salts. The
product was dissolved in 7:1 toluene/nBuOH at 80.degree. C., hot
filtered, and recrystallized by cooling and stirring overnight.
ER-806064 (F-1b) was isolated in a 57%, five step overall yield
after filtration and a toluene rinse. FAB(+)-MS m/z 357 (M+H).
Melting point 96.2.degree. C.
##STR00062##
Purified triol ER-806064 (1 wt., 1 eq.) was dispersed in acetone (2
vol.), diluted with 2,2-dimethoxypropane (1 vol.), and treated with
conc. sulfuric acid (0.0086 wts, 0.03 eq.) at 25.degree. C. The
reaction was stirred until homogenous. The reaction was diluted
with toluene (5 vol.) and quenched by the addition to 5%
K.sub.2CO.sub.3 (2 vol.). The layers were mixed well and separated.
The organic layer was washed with 10% brine, dried with
Na.sub.2SO.sub.4 (0.5 wt.). The solution was filtered (toluene
rinse) and concentrated at reduced pressure to afford ER-806126 as
a yellow oil. The material was used as is in the next stage.
##STR00063##
Solid NaOtBu (0.34 wt., 1.4 eq.) was dissolved in THF (2.7 vol.)
and DMF (0.3 vol.), and then cooled below 10.degree. C. A solution
of ER-806126 (1 wt., 1 eq.) in THF (2.5 vol.) was added to the
NaOtBu solution with a THF rinse (0.5 vol.), keeping the internal
temperature below 15.degree. C. After a 30 minute stir, methyl
iodide (0.204 vol., 1.3 eq.) was added keeping the temperature
below 15.degree. C. (exothermic). The reaction was warmed to
25.degree. C. and the reaction quenched with water (5 vol.) and
diluted with toluene (7 vol.). The layers were mixed well and
separated. The organic layer was washed twice with brine (2.times.5
vol.), dried over Na.sub.2SO.sub.4 (0.5 wt.), filtered, and
concentrated under reduced pressure.
##STR00064##
ER-806068 (1 wt., 1 eq.) was dissolved in 1 vol. of MeOH. Water
(1.5 vol.) and 2 N HCl (1.25 vol., 1 eq.) were added and the
reaction stirred at 25.degree. C. The reaction was quenched by
inverse addition to 2M NaOH (1.34 vol.) at 10.degree. C. The
reaction was diluted with isopropyl acetate (5 vol.), the layers
were mixed well and separated. The aqueous layer was back extracted
with 5 vol. of isopropyl acetate and the combined organic layers
were dried over MgSO.sub.4 (0.5 wt.), filtered, and concentrated at
reduced pressure to afford crude diol ER-806063.
##STR00065##
To a 25.degree. C. solution of crude ER-806063 (1 wt., 1.0 eq.) in
DMF (4 vol.), was charged imidazole (0.62 wt., 3.4 eq.), followed
by TBSCl (1.02 wt., 2.53 eq.) with the internal temperature below
30.degree. C. The reaction was stirred at 25.degree. C. The
reaction was diluted with MTBE (10 vol.) and washed with H.sub.2O
(4 vol.). The organic layer was washed sequentially with 1M HCl (3
vol.), water (3 vol.), aqueous sodium bicarbonate (3 vol.), and
brine (3 vol.). The organic layer was dried over MgSO.sub.4 (0.5
wt.), filtered with one volume MTBE rinse, and concentrated at
reduced pressure and solvent exchanged for heptane (4 vol.).
##STR00066##
ER-806065 (1 wt., 1 eq.) was dissolved in heptane, isooctane, or
IPA (10 vol.) The solution was cooled below -60.degree. C.
(.+-.10.degree. C.). Ozone was bubbled through the solution at low
temperature until the solution retained a blue color. Nitrogen was
purged through the solution for 15-30 minutes, and the reaction
warmed to 5.degree. C. while the nitrogen flush was continued. 7-15
wt. % Lindlar Catalyst (5% Pd on CaCO.sub.3 poisoned with Pb, 0.1
wt.) was added. The reactor head was purged several times with
nitrogen, evacuated, and placed under 1 atmosphere H.sub.2 (g). The
reaction was then warmed to room temperature (20-25.degree. C.).
The reaction was stirred for 2.5 hours. The resulting heterogeneous
solution was filtered through Celite.RTM. (1.0 wt.) with an MTBE (2
vol.) rinse. The solution was concentrated to dryness, isolating
1.0 wt. of crude ER-806067. The crude isolate was recrystallized
from heptane or isooctane to afford ER-806067 (F-1a) as a white
crystalline solid in a 68% five step yield. FAB(+)-MS m/z 601
(M+H). Melting point 64.5.degree. C.
Example 2
Preparation of F-2a
##STR00067##
A reactor was charged with pre-rinsed Amberlyst 15 (0.05 wt.) and
water (4.63 vol.) and cooled to an internal temperature of
0-5.degree. C. The reactor was charged with 2,3-dihydrofuran (1
wt., 1 eq.) and stirred for 1.5 hours maintaining internal
temperature around 5.degree. C. A second reactor was charged with
water (4.63 vol.) and heated to an internal temperature of
35.degree. C. The same reactor was charged with tin powder (2.2
wt., 1.3 eq.), distilled 2,3-dibromopropene (3.71 wt., 1.3 eq.),
and 48% hydrobromic acid (0.002 vol.), respectively. After
observation of reaction initiation indicated by a temperature spike
to 36-38.degree. C., the second reactor was charged with
2,3-dibromopropene portion-wise (9.times.0.37 wt.) while
maintaining the internal temperature below 45.degree. C. After
complete addition, the contents of the second reactor were stirred
at an internal temperature of 35.degree. C. for an additional 60
minutes. The filtered contents of the first reactor were charged
into the second reactor at a rate such that internal temperature
did not exceed 45.degree. C. After complete addition, the heat
source was removed and the second reactor was charged with
Celite.RTM. 545 (2.0 wt.) and the resulting mixture stirred for 30
minutes. The heterogeneous mixture was filtered through a
Celite.RTM. 545 pad (2.0 wt.) and the cake washed with additional
water (5 vol.). All filtrates were combined into a reactor and
charged with concentrated hydrochloric acid (1.5 vol.) until the
cloudy solution becomes clear. With vigorous stirring, the reactor
was charged with sodium chloride (3.6 wt.) and the layers allowed
to partition. The organic layer was separated and set aside. The
aqueous layer was extracted with n-butanol (20 vol.). The aqueous
layer was drained and the reactor charged with the organics from
the first separation. The organics were washed with concentrated
sodium bicarbonate (24 vol.), followed by a back extraction of the
aqueous layer with n-butanol (20 vol.). All organics were combined
and concentrated in vacuo. The concentrate was dissolved in MTBE
(10 vol.), filtered and the filtrate was concentrated to two vol.
With stirring, the concentrate was cooled to an internal
temperature of 0.degree. C. and then n-heptane (4 vol.) was added.
The heterogenous mixture was stirred for 2 hours at an internal
temperature of 0.degree. C. and the desired product was isolated
via filtration and dried under vacuum to yield ER-806909 (1.34 wt.,
0.45 eq.) as a white powder.
##STR00068##
A reactor was charged with imidazole (0.65 wt., 2 eq.), ER-806909
(1 wt., 1 eq.), and anhydrous DMF (4.04 vol.). With stirring, the
reactor was cooled to an internal temperature of 0.degree. C. and
then with tert-butylchlorodiphenylsilane (1.25 wt., 0.95 eq.) at a
rate such that internal temperature did not exceed 15.degree. C.
While maintaining the internal temperature <15.degree. C., the
reaction was stirred for an additional 1 hour. The reactor was
charged with water (3.2 vol.) and n-heptane (6.4 vol.). The mixture
was stirred for 5-15 minutes and the layers allowed to separate.
The organic layer was separated and set aside. The aqueous layer
was extracted with n-heptane (3.2 vol.). All organics were combined
and washed with brine (3.2 vol.) and concentrated in vacuo to a
constant weight to yield ER-806545 (2.13 wt., 0.95 eq.) as a yellow
oil. The product was utilized in the next stage without further
purification.
##STR00069##
The enantiomers of ER-806545 were separated via Simulated Moving
Bed (SMB) chromatography to yield ER-808373 (0.55 wt., 0.55 eq.)
and ER-806721 (0.45 wt., 0.45 eq.) as yellow oils. The SMB
chromatography protocol used to separate the enantiomers of
ER-806545 is as follows.
TABLE-US-00001 Columns and media: Chiracel OD 20 .mu.m 30 mm
.times. 150 mm (12 columns) Solvent system: 96:4 (vol/vol)
heptane:tert-butanol (mobile phase) Simulated Moving Knauer SMB
System CSEP C912 Bed Chromatography apparatus: Isotherms
(Langmurian, determined by frontal analysis): Undesired isomer: Qi*
= 2.8768 .times. Ci/(1 + 0.02327 .times. Ci) Desired isomer: Qi =
4.5320 .times. Ci/(1 + 0.0034595 .times. Ci) *Where Qi = solid
phase concentration (in g/L) and Ci = liquid phase con- centration
(g/L). Column porosity: 0.658 Temperature: 27.degree. C.
Flow rates, etc. calculated by simulation on EuroChrom 2000 for
Windows, SMB Guide ver.1.2, Wissenschaftliche Geratebau Dr.-Ing.
Herbert Knauer GmbH, D-14163 Berlin; authors, H. Kniep and A.
Seidel-Morgenstern:
TABLE-US-00002 Feed concentration: 36 g/L (ER-806545) Feed flow
rate (Pump 1): 15 mL/min Eluent flow rate (Pump 2): 76.4 mL/min
Zone IV (Pump 3) flow rate: 107 mL/min Zone II (Pump 4) flow rate:
134.3 mL/min (actual flow rate = 143.5 mL/min) Zone I flow rate:
183.4 mL/min Zone III flow rate: 149.3 mL/min Raffinate* flow rate:
42.3 mL/min *Raffinate = weakly bound isomer Extract flow rate:
49.1 mL.min *Extract = strongly bound isomer Tact time 0.8864 min
(53.18 sec, actual tact (port switching time): time = 54 sec)
The enantiomers of ER-806545 were separated using the above
protocol in the following manner. For an 11 hour run, 10 L of 36
g/L ER-806545 in mobile phase was pumped (Silog model Chemtech)
through a 142 mm diameter 0.45 .mu.m pore size nylon filter
(Cole-Parmer # 2916-48) and into the Feed tank. The Eluant tank was
filled with 36L mobile phase that has been filtered through an
in-line 45 mm diameter 1 .mu.m glass fiber filter (Whatman GFC),
additional vol. were added throughout the run. The internal
temperature of the SMB apparatus was adjusted to 27.degree. C.
For initial startup, the Feed and Eluant inlets were both connected
to the Eluant tank. The Feed and Eluant pumps were primed and
purged with mobile phase solvent. The SMB apparatus column
switching was initiated, the pumps were turned on and the flow
rates were gradually increased to full speed while maintaining
absolute flow rate differences between each pump. Once fall speed
was achieved the Raffinate and Extract flow rates were measured and
adjustments to pump flow rates were made to correct for deviations
in pump specifications. The Feed pump (Pump 1) was reduced to 0
mL/min, the inlet reconnected to the Feed tank, the pump primed
with Feed solution and then the flow rate gradually increased back
to full operating speed. The Raffinate and Extract outlets were
collected into separate tanks and samples of each of each were
acquired every 2 hours. The samples were monitored for chiral
purity by analytical HPLC using the HPLC method set forth below.
Adjustments to the flow rates of Pumps 2, 3 and 4 as well as to the
tact time were made to afford the desired outlet purities.
At the end of the run, the Feed pump was once again reduced to zero
flow rate and connected to the Eluant tank. The Feed pump was
brought back to full speed and the system was allowed to wash for
20 minutes. The Raffinate and Extract outlets were maintained for
10 minutes (10 tacts) during the wash period and, for the remainder
of the wash, the outlets were collected into a separate tank. The
column wash was eventually concentrated and added to the Feed on
subsequent runs.
The collected Extract (ER-806721) at the end of each run was pooled
with material collected from the same starting material lot and the
final pooled lot was analyzed again for chiral purity by the
analytical HPLC method described in Table 1 below. The same
procedure was applied to the collected Raffinate (ER-808373).
TABLE-US-00003 TABLE 1 HPLC Analysis of ER-806721 chiral purity:
Column: Chiracel OD 10 um 250 .times. 4.6 mm, DAICEL Chemical
Industries, Ltd., cat. no. 14025 Flow rate: 0.8 mL/min Temp.
(.degree. C.): 27 preferred over 35 Inj. Vol.: 10 uL usually,
sample in Solvent A, 5 mg/mL Instrument: Waters Alliance W2690 with
UV W2487 (also with Advanced Laser Polarimeter) Mobile Phase
Constituents: (PDR-Chiral, Inc.) A 99:1 Heptane:2-Propanol B C D
Gradient Table: (%) Gradient Time (min) A B C D 0 100 0 0 0
isocratic Run time 30 min Detection: Absorbance at 254 nm UV
##STR00070## ##STR00071## ##STR00072##
A reactor was charged with triphenylphosphine (0.7 wt., 1.2 eq.),
p-nitrobenzoic acid (0.45 wt., 1.2 eq.), ER-808373 (1 wt., 1 eq.),
and anhydrous toluene (8 vol.). The reaction was cooled to internal
temperature of 0.degree. C. and DEAD (1.17 wt., 1.2 eq.) was slowly
added at a rate such that the internal temperature did not exceed
7.degree. C. n-Heptane (3.3 vol.) was added and the mixture cooled
to an internal temperature 10.degree. C. then stirred for 30-40
minutes. The resulting precipitate was removed by filtration. The
filter-cake was washed with n-heptane (3.3 vol.), TBME (0.55 vol.),
n-heptane (1.1 vol.), and MTBE (0.55 vol.), respectively. All
filtrates were combined and concentrated in vacuo. The crude
concentrate was dissolved in THF (8 vol.) then water (0.8 vol.) and
lithium hydroxide dihydrate (0.18 wt., 2 eq.) were added. The
mixture was stirred at ambient temperature then n-heptane (3.3
vol.) was added and stirred for 5 minutes. Water (2.2 vol.) and
n-heptane (3.3 vol.) were added, the biphasic mixture was stirred
for 5 minutes, and the layers were allowed to partition. The
aqueous layer was separated and back extracted with n-heptane as
necessary. The organic layers were combined and concentrated in
vacuo. The crude product was purified via SiO.sub.2 column
chromatography to yield ER-806721 (0.74-0.85 wt., 0.74-0.85 eq.) as
a light yellow oil.
##STR00073##
A reactor was charged with ER-806721 (1 wt., 1 eq.) and anhydrous
dichloromethane (4.2 vol.). The reaction was cooled to an internal
temperature of 0-5.degree. C., then triethylamine (0.34 wt., 1.5
eq.), p-toluenesulfonyl chloride (0.51 wt., 1.2 eq.), and
4-(dimethylamino)-pyridine (0.001 wt., 0.25 eq.) were added. The
resulting mixture was stirred at ambient temperature for 48 hours
then water (1.8 vol.) and dichloromethane (1.8 vol.) were added.
After sufficient mixing, the organics were separated and
concentrated. The concentrate was dissolved in MTBE (1.8 vol.) and
washed with brine (1.8 vol.). The organic layer was separated and
set aside. The aqueous layer was back extracted with MTBE (1.8
vol.) then all organics were combined and concentrated in vacuo.
The crude oil was filtered through a plug of SiO.sub.2 (70-230
mesh, 1 wt.) eluting with MTBE (7 vol.) and the filtrates were
concentrated in vacuo. The concentrate was dissolved in IPA (5
vol.) and water (0.25 vol.) was added. The resulting mixture was
cooled to an internal temperature of 15.degree. C. and then seeded
with ER-807204. After seeding, the mixture was cooled to an
internal temperature of 0.degree. C. and stirred for 4-5 hours. The
suspension filtered, the filter cake was washed with cold IPA (1
vol.), and the cake dried in vacuo to a constant weight to yield
ER-807204 (1.05 wt., 0.78 eq.) as a white powder. IR (thin film,
cm-1) .lamda. 2597, 1633, 1363, 1177, 907, 729. LRMS m/z 602
(M+H).
##STR00074##
A reactor was charged with 21% sodium ethoxide in ethanol (2.97
wt., 0.9 eq.). The solution was heated to an internal temperature
of 65.degree. C. then diethyl malonate (3.24 wt., 2 eq.) was added
at a rate such that the internal temperature did not exceed
70.degree. C. The mixture was stirred for 30 minutes and then
ER-806906 (1 wt., 1 eq.) was added over 3-5 hours. Upon complete
addition, the reaction was stirred for 60 minutes and then cooled
to an internal temperature of 50.degree. C. Concentrated
hydrochloric acid (0.84 wt., 1.05 eq.) was added at a rate such
that internal temperature did not exceed 65.degree. C. The DMF (3
vol.) and ethanol were removed via distillation then a solution of
magnesium chloride hexahydrate (0.21 wt., 0.1 eq.) in distilled
water (0.25 vol., 1.4 eq.) was added. The resulting mixture was
heated to an internal temperature of 135.degree. C. while removing
the distillate. The mixture was heated at reflux then cooled to
room temperature and brine (12 vol.) and TBME (16 vol.) were added.
The organic layer was separated and washed with water (1.3 vol.)
and brine (1.2 vol.) then concentrated in vacuo. The product was
purified via distillation to yield ER-805552 (0.95-1.09 wt., 0.71
eq.).
##STR00075##
A reactor was charged with LHMDS 1.0 M in toluene (6.61 wt., 1.04
eq.) and cooled to an internal temperature of -75.degree. C.
ER-805552 (1 wt., 1 eq.) was dissolved in anhydrous THF and added
to the reactor at a rate such that internal temperature did not
exceed -70.degree. C. Upon complete addition, the resulting mixture
was stirred for 30 minutes. A second reactor was charged with
anhydrous THF (2.5 vol.) and methyl iodide (1.27 wt., 1.25 eq.) and
cooled to an internal temperature of -75.degree. C. The solution of
ER-805552 in THF was added into the methyl iodide solution at a
rate such that internal temperature did not exceed -65.degree. C.
Upon complete addition, the reaction was stirred at an internal
temperature of -78.degree. C. for 30 minutes. The reaction was
inverse quenched a solution of 1 N hydrochloric acid (10 vol.) and
MTBE (8 vol.) with vigorous stirring. After complete addition, the
aqueous layer was separated and discarded. The organic layer was
washed with brine solution (3 vol.) and concentrated in vacuo and
the product purified via distillation to afford ER-806724 (0.75
wt.) as a .about.6/1 mixture of diastereomers.
##STR00076##
A reactor was charged with N,O-dimethylhydroxylamine HCl (1.05 wt.,
1.5 eq.) and anhydrous CH.sub.2Cl.sub.2 (8.1 vol.) and cooled to an
internal temperature of 0.degree. C. 2 M trimethylaluminum in
toluene (3.93 wt., 1.5 eq.) was added at a rate such that internal
temperature did not exceed 5.degree. C. The reaction was stirred
for an additional 10 minutes and ER-806724 was added at a rate such
that internal temperature did not exceed 5.degree. C. The reaction
was diluted with CH.sub.2Cl.sub.2 (15 vol.) then inverse quenched
into 1.3 M sodium tartrate (20 vol.) at an internal temperature of
0.degree. C. at a rate such that the internal temperature did not
exceed 10.degree. C. After complete addition, the layers were
partitioned and the aqueous layer was separated and set aside. The
organics were washed with water (1 vol.), dried over sodium sulfate
(1 wt.), filtered and concentrated in vacuo until minimal methylene
chloride was being removed. To the concentrate was added anhydrous
DMF (6.3 vol.), imidazole (0.64 wt., 1.5 eq.) and
t-butyldimethylsilyl chloride (0.94 wt., 0.97 eq.), respectively.
Water (5 vol.) and MTBE (10 vol.) were added and the resulting
mixture stirred then the layers were allowed to partition. The
aqueous layer was separated and discarded. The organic layer was
washed with water (5 vol.) and the layers separated. 1 N sodium
hydroxide (2.5 vol.) and methanol (2.5 vol.) were added and the
resulting mixture stirred. The aqueous layer was separated and the
organic layer washed with brine (2.5 vol.) then concentrated in
vacuo to afford ER-806753 (1.94 wt., 0.91 eq.) as a brown oil.
##STR00077##
A reactor was charged with ER-806753 (1 wt., 1 eq.),
CH.sub.2Cl.sub.2 (5 vol.) and NMO-50% in water (0.8 wt., 1.1 eq.).
The mixture was cooled to an internal temperature of 10.degree. C.
and then 0.197 M OsO.sub.4 in toluene (0.06 vol., 0.004 eq.) was
added. Sodium sulfite (0.1 wt., 0.25 eq.) and water (0.85 vol.)
were added and the reaction stirred for 1 hour. The mixture was
diluted with brine (0.85 vol.) and the organics were concentrated
in vacuo to approximately 1/3 vol. A second reactor was charged
with sodium periodate (1.3 wt., 2 eq.) followed by THF (2.5 vol.).
The mixture was diluted with pH=7 phosphate buffer (3.0 vol.) and
cooled to an internal temperature of 20.degree. C. The concentrated
diol was added at a rate such that the internal temperature did not
exceed 30.degree. C. After complete addition, the resulting mixture
was stirred at room temperature. Water (1.25 vol.), MTBE (7 vol.)
and brine solution (1.25 vol.) were added and the layers separated.
The organics were washed a second time with a mixture of brine
solution (1 vol.) and saturated sodium bicarbonate (1 vol.).
Finally, the organics were stirred over a mixture of brine (1 vol.)
and 10% (w/v) sodium thiosulfate solution (1 vol.) for 1 hour then
concentrated in vacuo. The crude material was purified via
SiO.sub.2 column chromatography to yield ER-806754 (0.93 wt., 0.93
eq.) as a yellow oil. IR (thin film, cm-1) .lamda. 2953, 2856,
1725, 1664, 1463, 1254, 1092, 833. LRMS m/z 332 (M+H).
##STR00078##
A reactor was charged with ER-806629 (1.53 wt., 3.1 eq.) and THF
(10.5 vol.) and the solution was degassed with nitrogen sparge for
60 minutes. A second inserted reactor was charged with ER-807204 (1
wt., 1.0 eq.), ER-806754 (0.66 wt., 1.2 eq.) and THF (2.7 vol.) and
this solution was degassed with argon sparge for 45 minutes. The
reactor containing ER-806629 was charged with CrCl.sub.2 (0.63 wt.,
3.1 eq.) and followed by Et.sub.3N (0.52 wt., 3.1 eq.). The dark
green suspension was stirred at an internal temperature of 30 to
35.degree. C. for 1 hour, cooled to 0 to 5.degree. C. and then
NiCl.sub.2 (0.1 eq.) was added. The first reactor was charged with
the contents of the second reactor slowly over 0.5 hours and the
reaction allowed to warm to rt. The reaction was cooled to an
internal temperature of 0.degree. C. then ethylenediamine (1.0 wt.,
10 eq.) was added over 30 minutes and the reaction stirred at an
internal temperature of 25.degree. C. for at least 30 minutes. To
the reaction was added water (4 vol.), TBME (10 vol.) and n-heptane
(1 vol.) and the resulting mixture stirred for 15 minutes and the
phases allowed to separate (.about.30 min). The aqueous phase was
separated and back extracted with TBME (.about.7.5 vol.). The
organic layers were combined and washed with water (5 vol.), brine
(3 vol.), and concentrated in vacuo to minimum volume. To the crude
mixture was added IPA (10 vol.) and SiO.sub.2 (1 wt.) and the
resulting mixture stirred at an internal temperature of 25.degree.
C. for up to 4 days. The slurry was filtered and the filter cake
washed with IPA (2.times.1 volume). To the filtrate was added
n-heptane (6.6 vol.) and the mixture was concentrated in vacuo
until a suspension formed. The mixture was filtered and the cake
washed with n-heptane then the mixture was concentrated in vacuo.
The crude product was purified via SiO.sub.2 column chromatography
to yield ER-807524 (0.54 wt., 0.48 eq.), as a clear yellow oil. IR
(thin film, cm-1) .lamda. 2934, 1668, 1471, 1108, 833. LRMS m/z 704
(M+Na).
##STR00079##
A reactor was charged with ER-807524 (1 wt., 1 eq.) and anhydrous
THF (1.25 vol.). The mixture was cooled to an internal temperature
of -20.degree. C. and 3 M methyl magnesium chloride (0.59 vol., 1.2
eq.) was added at a rate such that the internal temperature did not
exceed 0.degree. C. Upon complete addition, the mixture was warmed
to an internal temperature of 0.degree. C. over 2 hours. The
reaction mixture was inverse quenched into semi-saturated ammonium
chloride (2.62 vol.) and the resulting mixture diluted with TBME (2
vol.) with vigorous mixing. The aqueous layer was discarded and the
organics washed with brine (2 vol.) then concentrated in vacuo. The
crude product was purified via SiO.sub.2 column chromatography to
yield ER-807525 (0.79-0.82 wt., 0.85-0.88 eq.) as yellow oil.
##STR00080##
A reactor was charged with ER-807525 (1 wt., 1 eq.),
N-phenylbistrifluoromethanesulfonamide (0.59 wt., 1.1 eq.), and
anhydrous THF (4.1 vol.) and the mixture cooled to an internal
temperature of -75.degree. C. 0.5 M KHMDS in toluene (2.75 wt., 1
eq.) was added at a rate such that internal temperature did not
exceed -60.degree. C. then the reaction was warmed to -20.degree.
C. over 2 hours. The reaction was quenched with semi-saturated
NH.sub.4Cl (2.4 vol.) at a rate such that internal temperature did
not exceed 0.degree. C. The mixture was warmed to an internal
temperature of 20.degree. C. and n-heptane (2.4 vol.) was added.
The mixture was stirred and the aqueous layer was separated and
discarded. The organic layer was washed three times with saturated
sodium bicarbonate (2.3 vol. each) then concentrated in vacuo to
yield ER-807526 (1.2-1.4 wt., 1.0-1.2 eq.). The material was
utilized in the next stage without further purification.
##STR00081##
A reactor was charged with ER-807526 (1 wt., 1 eq.) and anhydrous
methanol (3 vol.) at 20.degree. C. 1.25M HCl in IPA (4 vol., 5 eq.)
was added and the mixture stirred for 3 hours. Solid NaHCO.sub.3
(0.42 wt., 5 eq.) was added portion-wise with stirring until the pH
of the reaction mixture reached 6-7. The reaction mixture was
filtered with methanol (3.times.2 vol.) washes. All filtrates were
concentrated in vacuo and then purified via SiO.sub.2 column
chromatography to yield ER-807527 (0.43 wt., 0.79-0.85 eq.).
##STR00082##
The diastereomeric mixture of ER-807527 was separated by
preparative HPLC chromatography and the desired fractions
concentrated to yield ER-806730 (0.56 wt., 0.56 eq.) as a clear
yellow oil. The preparative HPLC chromatography protocol used to
isolate ER-806730 is as follows.
TABLE-US-00004 Column and Media: Kromasil spherical silica 60
.ANG., 10 .mu.m particle size packed to 7.7 cm (diam.) .times. 30
cm (length) in a 7.7 cm .times. 60 cm Varian Dynamax Rampak column.
HPLC Packing Station: Varian (Rainin) Dynamax Rampak 41/77 mm
Column Packing Station HPLC Pumps: Varian (Rainin) SD-1 Titanium
Pump Heads Primary HPLC Detector: Waters R403 Refractive Index
detector Secondary HPLC Detector: Varian (Rainin) UV-1 detector
with preparative flow cell Chromatography Control Varian (Rainin)
Dynamax DA version 1.4.6 and Acquisition Software: Chromatography
Data Varian (Rainin) Dynamax R version 1.4.3 Processing Software:
Mobile Phase: 28.5:63.7:7.85 (vol/vol) n-heptane: methyl tert-butyl
ether:2-propanol Flow Rate: 140 mL/min Column Temperature: Ambient
(25.degree. C.) Detection: Refractive Index, negative polarity at
16 X affenuation and UV at 215 nm. Mobile Phase Gradient: Isocratic
Run Time: 40 minutes Injection Volume: 10 ml of 0.8 g/mL of
ER-807527
The above protocol was used to separate the diastereomers of
ER-807527 in the following manner. Each lot of ER-807527 was first
diluted to 0.1 g/ml in the mobile phase and filtered under vacuum
through a 47 mm, 1 .mu.m pore size, glass fiber filter (Whatman
GFC). The filtrate was then concentrated under vacuum on a rotary
evaporator. Flow on the SD-1 HPLC pump A (primed and purged with
mobile phase) was initiated and the flow rate gradually increased
to 140 mL/minute. The system was washed until the UV and RI
detectors achieved a stable baseline. The RI detector reference
flow cell was flushed with fresh mobile phase.
Chromatography of 8 g injections of ER-807527 was accomplished by
diluting the current lot of ER-807527 to a concentration of 0.8
g/mL in the mobile phase. Injecting 10 mL aliquots of the dissolved
material and collecting the eluant corresponding to the ER-806730
peak approximately beginning at the peak apex approximately 24
minutes and continuing to 35 minutes. Subsequent injections and
fraction collection were continued until the starting material is
exhausted.
The fractions corresponding to ER-806730 were pooled and
concentrated under vacuum on a rotary evaporator. The
diastereomeric purity and area-% purity area were assessed using
the HPLC analytical method described in Table 2.
TABLE-US-00005 TABLE 2 HPLC Analysis of Diastereomeric Purity of
ER-806730: Column: Kromasil .[.Slica.]. .Iadd.Silica .Iaddend.250
.times. 4.6 mm, 5 .mu.m, MetaChem cat. no. 0475-250X046 Flow rate:
1 mL/min Temp. (.degree. C.): 27 Inj. Vol.: 10 .mu.L, sample in
Solvent A, 2.5 mg/mL Instrument: Waters Alliance W2690 with UV
W2487 Mobile Phase Constituents: A 30:67:3 n-Heptane:Methyl
tert-Butyl Ether:2-Propanol B 2-Propanol C D Gradient Table: (%)
Gradient Time (min) A B C D 0 100 0 0 0 isocratic 22 100 0 0 0
isocratic 26 90 10 0 0 linear 32 90 10 0 0 isocratic Run time 32
min with 18 min re-equilibration time at initial conditions
Detection: Absorbance at 205 nm UV ##STR00083## ##STR00084##
A reactor was charged with ER-806730 (1 wt., 1 eq.) and anhydrous
dichloromethane (4.8 vol.) and cooled to an internal temperature of
0.degree. C. 2,4,6-Collidine (1.16 wt., 4 eq.) and DMAP (0.03 wt.,
0.1 eq.) were added and the resulting mixture stirred for 15
minutes and then trimethylacetyl chloride (0.3 wt., 1.05 eq.) was
added at a rate such that internal temperature did not exceed
10.degree. C. Water (3 vol.) was added and the mixture stirred for
15 minutes. TBME (10 vol.) was added and the mixture stirred for an
additional 10 minutes. The organic layer was washed with 1N HCl (10
vol.) washing until a negative result for 2,4,6-collidine is
obtained then with water (5 vol.), saturated sodium bicarbonate (5
vol.), and saturated brine (5 vol.), respectively. The organic
layer was concentrated in vacuo and the concentrate purified via
SiO.sub.2 column chromatography to yield ER-806732 (1.02 wt., 0.85
eq.) as a yellow oil.
##STR00085##
A reactor was charged with ER-806732 (1 wt., 1 eq.) and anhydrous
THF (2.35 vol.) and cooled to an internal temperature of 0.degree.
C. Triethylamine (0.22 wt., 1.1 eq.) was added followed by
methanesulfonyl chloride (0.24 wt., 1.05 eq.) at a rate such that
internal temperature did not exceed 10.degree. C. The reaction was
stirred at an internal temperature of 0.degree. C. then n-heptane
(3.4 vol.) was added with vigorous stirring and the layers were
allowed to partition. The organics were washed with saturated brine
(3.4 vol.), dried over saturated sodium sulfate (2 wt.), filtered
and the cake washed with n-heptane until a negative result for
ER-805973 (F-2a) was obtained. The filtrates were concentrated in
vacuo to obtain ER-805973 (1.12 wt., 0.97 eq.). The crude ER-805973
(F-2a) was used in the next stage without further purification. IR
(thin film, cm-1) .lamda. 2961, 1725, 1413, 1208, 926. LRMS m/z 579
(M+H).
Example 3
Alternate Preparation of ER-806730
##STR00086##
Quinic acid (1 wt), cyclohexanone (2.11 eq, 1.08 wt), and conc.
sulfuric acid (0.011 eq, 0.0056 wt) were added to a reactor. The
reaction mixture was heated to 160.degree. C. and water was removed
by azeotropic distillation (azeotrope begins at 100.degree. C.).
The reaction was cooled to 90.degree. C. to 100.degree. C. and
sodium bicarbonate (0.0096 wt) and toluene (3.6 wt) were added. The
reaction was cooled to ambient temperature over 4-6 hours and the
resulting precipitate was filtered, washed with toluene
(2.times.0.9 wt), and dried to provide 1 (0.97 wt) as a white
powder.
##STR00087##
Compound 1 (1 eq, 1 wt) and imidazole (2.5 eq, 0.80 wt) were
combined, purged with N.sub.2, and suspended in anhydrous THF (10
v). TMSCl (1.2 eq, 0.61 wt) was added at a rate that maintained the
temperature below 30.degree. C. The reaction was cooled to ambient
temperature, heptane (10 v) was added and the resulting suspension
filtered. The filter cake was washed with 1:1 hpt/THF (10 v) the
filtrate solvent was exchanged with toluene by atmospheric
distillation to provide a solution of 2a (calcd. at 1.34 wt) in
toluene (.about.5 wt).
The solution of 2a was cooled to -78.degree. C. and DIBAL-H (1.5 M
in toluene, 1.2 eq, 2.1 wt) was added at a rate that maintained the
temperature below -65.degree. C. The excess DIBAL-H was quenched
with MeOH (0.3 eq, 0.034 wt) and the solution warmed to 0.degree.
C. The solution was transferred to a solution of 30% wt/wt aqueous
Rochelle salt (10 wt) and sodium bicarbonate (1 wt) at a rate that
maintained the temperature below 25.degree. C. The mixture was
stirred vigorously to obtain a biphasic solution. The layers were
separated and the aqueous layer was extracted with MTBE (5v). The
combined organic layers were washed with water (2.5 wt) and then
saturated brine (2.5 wt). The organics were concentrated and
solvent exchanged with THF to provide a solution of 2b (calcd. at
0.98 wt) in THF (5 v).
The solution of 2b was cooled to 5.degree. C. and acetic acid (2.9
eq, 0.51 v) was added. Water (1.0 eq, 0.055 v) was added and the
solution stirred at 0.degree. C. to 5.degree. C. Up to two
additional aliquots of acetic acid and one aliquot of water were
added as needed to facilitate deprotection of the silyl group.
Et.sub.3N (12 eq, 3.6 wt) and DMAP (0.05 eq., 0.02 wt) were added
at a rate that maintained the temperature below 20.degree. C.
Acetic anhydride (6 eq, 2.0 wt) was added and the reaction stirred
at rt. The reaction was cooled to 5.degree. C. and added to
saturated aqueous sodium bicarbonate (10 v) at a rate that
maintained the temperature below 30.degree. C. The resulting
mixture was allowed to stir for 3-4 hours and the layers allowed to
separate. The aqueous layer was extracted with MTBE (5 v) and the
combined organics were washed with water (5 v). The extracts were
solvent exchanged with IPA by distillation to provide a solution of
2c in IPA (3 v). The solution was cooled to 5.degree. C. and the
resulting crystals were filtered. The mother liquor was
concentrated and a second crop obtained after recrystallization to
provide 2c (0.87 wt) as a white crystalline solid.
##STR00088##
2c (1 wt) was dissolved in acetonitrile (6 v) and methyl
3-trimethylsilylpent-4-eneoate (3.0 eq, 1.86 wt) was added followed
by TFAA (0.2 eq, 0.083 v). BF.sub.3OEt.sub.2 (1.0 eq, 0.42 v) was
then added to the solution at a rate that maintained the
temperature below 25.degree. C. The reaction was added to saturated
aqueous sodium bicarbonate (10 v) and the resulting mixture stirred
for 15 minutes. The mixture was extracted with heptane (10 v),
followed by MTBE (5 v) and the combined extracts were concentrated
to provide 3 (calcd. at 0.72 wt) as an orange oil.
##STR00089##
A solution of 3 (1 wt) in THF (9 v) was treated with sodium
methoxide (25% wt/wt in methanol (1.5 eq, 2.2 wt)) at a rate that
maintains the temperature below 25.degree. C. The reaction was
quenched by addition to 1 N HCl (10 v). The organic layer was
separated and the aqueous was extracted with MTBE (10 v). The
combined organics were washed with water (2.5 v), saturated sodium
bicarbonate (2.5 v), and water (2.5 v). The solution was
concentrated to provide a solution of 4 (calcd. at 0.88 wt) in THF
(2.5 v). The solution was used directly in the next step.
##STR00090##
Methanol (5 v) was added to the solution of 4 (1 wt) in
tetrahydrofuran (2.5 v). 1N HCl (0.75 eq, 2 v) was added and the
reaction was warmed to 60-80.degree. C. The reaction was cooled to
rt and added to saturated aqueous bicarbonate. The mixture was
extracted with DCM (3.times.2.5 v) and the combined DCM extracts
were solvent exchanged with EtOAc to provide a solution of 5 in
EtOAc (3 v). Heptane (2 v) was added to induce crystallization and
the resulting suspension cooled to 0.degree. C. The solids were
collected by filtration and the filter cake washed with cold
EtOAc/heptane (1:1 v/v) and dried to provide 5 (0.55 wt) as a white
powder.
##STR00091##
Compound 5 (1 wt) was dissolved in ACN (10 v) then
2-acetoxy-2-methylpropanyl bromide (4.0 eq, 2.2 wt) and water (1
eq, 0.067 wt) were added consecutively. The resulting mixture was
stirred at ambient temperature then cooled to 5-10.degree. C. NaOMe
(25% wt/wt in MeOH, 8 eq, 6.2 wt) was added and the reaction
allowed to warn to ambient temperature. The reaction was quenched
by the addition of saturated sodium bicarbonate (10 v) and
extracted with MTBE (2.times.10 v). The solvent was exchanged with
methanol by atmospheric distillation to provide a solution of 6
(calcd at 0.91 wt) in methanol (10 v).
##STR00092##
A solution of 6 (1 wt) in methanol (10 v) was heated to 55.degree.
C. Sodium borohydride (5 eq, 0.68 wt) was added in 6 portions and
the reaction cooled to 5.degree. C. and quenched with 1 N HCl (10
v). Brine (5 v) was added and the reaction extracted with EtOAc
(2.times.10 v). The extracts were combined and concentrated to
provide 7 as a tan residue.
##STR00093##
Compound 7 (1 wt) was dissolved in CH.sub.2Cl.sub.2 (10 v) then
DMAP (0.1 eq, 0.054 wt), Et.sub.3N (3.0 eq, 1.85 v), and TBDP-SCl
(1.2 eq, 1.38 v) were added at ambient temperature. Sodium
bicarbonate (10 v) was added and the organic layer separated. The
aqueous layer was extracted again with CH.sub.2Cl.sub.2 (10 v), the
organic extracts combined and concentrated to provide 8 (calculated
at 1.8 wt) as a colorless oil.
##STR00094##
LDA (1.5 M in cyclohexane, 4 eq, 6 v) was added to a solution of 8
(1 wt) in THF (10 v) at ambient temperature. The solution was
warmed to 50.degree. C. then quenched with 1 N HCl (5 v) and
extracted with MTBE (10 v). The extracts were concentrated to
provide 9 (0.9 wt) as an oil.
##STR00095##
A solution of 9 (1 wt) in CH.sub.2Cl.sub.2 (5 v) in MeOH (5 v) was
cooled to -60.degree. C. and treated with O.sub.3 keeping
temperature below -50.degree. C. The reaction was purged with
N.sub.2, NaBH.sub.4 (0.5 eq, 0.04 wt) was added and the mixture
warmed to 0.degree. C. Additional NaBH.sub.4 (1 eq, 0.08 wt) was
added in portions and the reaction allowed to warm to ambient
temperature. After 3 hours, the mixture was quenched with 1N HCl,
(10 v), CH.sub.2Cl.sub.2 (5 v) was added and the layers were
allowed to partition. The aqueous layer was re-extracted with
CH.sub.2Cl.sub.2 (10 v) and the organic extracts were combined and
concentrated to provide 10 (0.97 wt) as a colorless oil.
##STR00096##
Compound 10 was dissolved in THF (10 v) and phosphate buffer (pH=7,
5 v) was added. NaIO.sub.4 (2 eq, 0.854 wt) was added and the
reaction warmed to ambient temperature. Water (5 v) and MTBE (10 v)
were added and the resulting mixture was stirred vigorously for 10
minutes. The organic layer was separated and washed with 10% wt/v
aqueous sodium thiosulfate (5 v), water (5 v), and brine (5 v) then
dried by azeotropic distillation with THF (.about.200 ppm water) to
provide a solution of 11 (calcd. at 0.93 wt) in THF (10 v). This
solution was used directly in next step.
##STR00097##
(Carbomethoxymethylene)triphenylphosphorane (1 wt) was added to the
solution of 11 (1 wt) in THF (10 v) and heated to 65.degree. C.
Heptane (40 v) was added and the resulting mixture stirred for 30
minutes. The resulting precipitate was filtered and the filtrate
concentrated to a total 10 v. SiO.sub.2 (5 wt) was added and the
suspension filtered over a pad of SiO.sub.2 eluting with MTBE
(20-40 v). The solvent was exchanged with MeOH to provide a
solution of 12 (calcd. at 0.95 wt) in MeOH (10 v) which was used
directly in the next step.
##STR00098##
A solution of 12 (1 wt) in MeOH (10 v) was added to 10% Wt/wt Pd(C)
(0.23 eq, 0.37 wt) and treated with H.sub.2. The suspension was
filtered while rinsing the filter cake with THF (10 v). The solvent
was exchanged with THF to provide a solution of 13 (calcd. at 0.95
wt) in THF (10 v) which was used directly in the next step.
##STR00099##
The solution of 13 (1 wt) in THF (10 v) was cooled to 0-5.degree.
C. and LAH (1 M (THF), 0.78 eq, 1.5 v) was added at a rate that
maintained the temperature below 10.degree. C. Water (1.7 eq, 0.06
v) was then added at a rate that maintained the temperature below
10.degree. C. NaOH (10% wt/wt in water, 0.16 eq, 0.06 v) was added
followed by water (4.98 eq, 0.17 v) at a rate that maintained the
temperature below 10.degree. C. and the resulting mixture stirred
vigorously while warming to ambient temperature. The suspension was
filtered and the filter cake rinsed with THF (5 v). The filtrate
was partially concentrated to provide 14 (calcd. at 0.9 wt) in THF
(10 v) which was used directly in next step.
##STR00100##
The solution of 14 (1 wt) in THF (10 v) was cooled to 0.degree. C.
then imidazole and TrCl (1.5 eq, 0.59 wt) were added. Saturated
aqueous NaHCO.sub.3 (5 v) was added and the mixture extracted with
heptane (10 v). The extract was washed with brine (10 v) and
concentrated to provide 15 (1.35 wt).
##STR00101##
Compound 15 (1 wt) was dissolved in THF (10 v) and treated with
TBAF (1M, 1.2 eq, 1.6 v). The reaction mixture was concentrated to
2 v then heptane (5 v) and SiO.sub.2 (5 wt) were added. The
resulting suspension was filtered and eluted with heptane (5 v)
followed by THF (10 v). The THF eluent was collected to provide a
solution of 16 (calcd at 0.61 wt) in THF (10 v) which was used
directly in the next step.
##STR00102##
PPh.sub.3 (5 eq, 2.3 wt), pyridine (10 eq 1 vol), and NIS (3.0 eq,
1.1 wt) were added to the solution of 16 (1 wt) in THF (10 v). 20%
wt/wt aqueous citric acid (10 eq, 14 wt) was then added and the
resulting mixture allowed to stir for 10 minutes. The reaction was
diluted with heptane (10 v) and the aqueous layer separated. The
organic layer was washed with water (5 v), 10% wt/v aqueous sodium
thiosulfate (5 v), water (5 v) and brine (5 v). The solvent was
exchanged with EtOH and concentrated to 5 v. Water (10 v) was added
and the resulting precipitate was collected by filtration to obtain
17 (0.65 wt) as a white solid.
##STR00103##
Compound 17 (1 wt) and KCN (6 eq, 0.54 wt) were suspended in EtOH
(5 v) and water (10 v) and the resulting suspension heated to
80.degree. C. The reaction was diluted with water (5 v) and EtOAc
(10 v) and mixed for 10 minutes. The aqueous layer was removed and
the organic layer washed with water (5 v) and brine (5 v). The
solvent was exchanged with EtOH to provide 18 (0.75 wt) in EtOH (10
v) which was used directly in next step.
##STR00104##
Zn (37 eq, 3.9 wt) was added to the solution of 18 (1 wt) in EtOH
(10 v) and the mixture heated to 75-80.degree. C. The reaction was
partially concentrated to 2-3 v, cooled to ambient temperature, and
partitioned between MTBE (10 v) and water 5 (v). The aqueous layer
was removed and the organic layer washed with saturated bicarbonate
(5 v), water (5 v), and brine (5 v), then dried by THF azeotropic
distillation to .about.200 ppm water to provide 19 (0.81 wt) in THF
(10 v). The resulting solution was used directly in next step.
##STR00105##
LDA (1.0 M in THF, 1.2 eq, 2.4 v) was added to the solution of 19
(1 wt) in THF (10 v) at -78.degree. C. The resulting mixture was
stirred for 10 minutes then the enolate solution was added to a
solution of MeI (1.5 eq, 0.19 v) in THF (5 v) at -78.degree. C. The
reaction was inverse quenched into saturated sodium bicarbonate (10
v) and extracted with MTBE (15 v). The extract was washed with
brine (5 v), concentrated, then purified by chromatography to
provide 20 (0.86 wt).
##STR00106##
AlMe.sub.3 (2 M in toluene, 1.5 eq, 1.5 v) was added to a
suspension of dimethylhydroxylamine hydrogen chloride (1 wt) in
CH.sub.2Cl.sub.2 (2.5 v) at 0.degree. C. A solution of 20 (1 wt) in
CH.sub.2Cl.sub.2 (5 v) was added at a rate that maintained the
reaction temperature below 5.degree. C. The reaction mixture was
then added to aqueous sodium tartrate (1.3 M, 20 v) keeping the
temperature below 10.degree. C. The layers were allowed to
partition, were separated, and the organic layer was dried with
Na.sub.2SO.sub.4 (5 wt). The resulting suspension was filtered and
the filtrate concentrated. The residue was dissolved in DMF (2 v)
then imidazole (0.19 wt) and TBSCl (0.29 wt) were added. The
reaction was diluted with water (5 v) and MTBE (10 v) and allowed
to stir for 10 minutes. The aqueous layer was removed and the
organic layer washed with water (5 v). The extract was added to a
solution of aqueous NaOH (1N, 0.78 v) and MeOH (0.7 v). The
reaction was allowed to stir then the aqueous layer was removed and
the organic washed with brine (2.5 v) then concentrated to provide
21 (1.2 wt).
##STR00107##
Methyl magnesium chloride (3.0 M, 59 wt, 1.2 eq) was is added to a
solution of 21 (1 wt) in anhydrous THF (1.11 wt, 1.25 v) at a rate
that maintained the reaction temperature below 0.degree. C. After
stirring at 0.degree. C., the reaction was reverse quenched into
saturated ammonium chloride (2.5 v) and water (2.3 v). The
resulting mixture was diluted with MTBE (10 v) and stirred
vigorously. The aqueous layer was separated and the organic layer
washed with brine (2.5 v) and concentrated to provide 22 (0.84
wt).
##STR00108##
Compound 22 (1 wt) was dissolved in THF (4 v) and cooled to
-78.degree. C. KHMDS (1.5 M in toluene, 1.01 eq, 2.78 wt,) was
added while maintaining the temperature below -60.degree. C. A
solution of Tf.sub.2NPh (0.62 wt, 1.1 eq) in THF (1.5 v) was added
and the reaction warmed to -20.degree. C. Saturated ammonium
chloride (2.5 v), water (2.5 v), and n-heptane (2.5 v) were added
and the mixture warmed to ambient temperature. The layers were
allowed to partition and the aqueous layer removed. The organic
extract was washed with saturated aqueous sodium bicarbonate
(3.times.2.5 v) and brine (2.5 v) then concentrated in vacuo to
provide 23 (1.1 wt).
##STR00109##
Compound 23 was dissolved in MeOH (2.5 v) and cooled to 15.degree.
C. HCl (5N in IPA, 1.30 eq, 1.18 wt) was added and the resulting
solution allowed to warm to 25.degree. C. The reaction was cooled
to 0.degree. C. and sodium bicarbonate (3 eq, 0.33 wt) was added.
The reaction was stirred for 15 minutes and the resulting
precipitate removed by filtration. The filter cake was washed with
ACS grade methanol (1 v) and filtrates were combined and
concentrated. The crude concentrate was purified by chromatography
to provide ER-806730 (24) (0.5 wt).
Example 4a
Example 4a provides an alternate method of preparing compounds of
formula A, an intermediate to F 2, using the general scheme set
forth at Scheme V above. This method uses ER-812935 as an
intermediate as prepared according to Example 3 (compound 4),
above.
##STR00110##
ER-812935 (1 wt) was dissolved in THF (10 v) and cooled to
0.degree. C. LAH (1.0 M in THF, 0.70 eq, 2.0 v) was added keeping
the temperature below 5.degree. C. While stirring vigorously,
excess reagent was quenched with water (0.078 v) keeping the
temperature below 5.degree. C. While maintaining the vigorous
stirring, NaOH (15% wt/wt in water (0.078 v)) was added followed by
water (0.18 v). After adding Celite.RTM. (2 wt), the suspension was
filtered and the cake rinsed with THF (5 v). The solution of
ER-817633 (0.92 wt, calcd. based on 100% conversion) was
concentrated to 5 v and used directly in the next stage.
##STR00111##
The previously prepared solution of ER-817633 (1 wt in 5 v THF) was
diluted with THF (5 v), cooled to 5.degree. C. and Et.sub.3N (3
eq., 0.94 wt) was added. MsCl (1.05 eq, 0.25 v) was added at a rate
that maintained the temperature below 10.degree. C. The reaction
was quenched by addition of water (5 wt). Heptane (8 v) was added
and the mixture allowed to partition. The aqueous phase was
separated and extracted with MTBE (2 v). The combined organic
extracts were washed with saturated sodium bicarbonate (5 v) and
water (1.9 v) The organic layer was concentrated and solvent
exchanged with EtOH to prepare a solution of ER-818937 (1.23 wt
calcd. based on 100% conversion) in EtOH (1 v) which was used
directly in the next stage.
##STR00112##
The previously prepared solution of ER-818937 (1 wt in EtOH (0.8 v)
is diluted with EtOH (190 proof, 9 v). KCN (3 eq., 0.41 wt) was
added and the suspension was heated to 70-80.degree. C. The
reaction was cooled to ambient temperature and water (10 v) was
added followed by MTBE (10 v). The layers were separated and the
aqueous extracted with MTBE (5 v). The combined organics are washed
with water (2 v) and saturated brine (4 wt). The extracts were
concentrated and used directly in the next stage.
##STR00113##
ER-818950 was dissolved in acetic acid (5 v) and hydrogen chloride
(1.0 M, 1 eq, 3 v) was added and the reaction was stirred at
ambient temperature. The reaction was cooled to 0.degree. C. and
NaOH (50% wt/wt, 30 eq, 7 wt) was added at a rate that maintained
the temperature below 10.degree. C. The solution was extracted with
heptane (2.times.10 vol). The aqueous phase was saturated with NaCl
and extracted with ACN (2.times.10 v). The combined ACN extracts
were concentrated and solvent exchanged with EtOAc by atmospheric
distillation to provide a solution of ER-817664 in EtOAc (3 v).
Salts were filtered from the hot solution which was then cooled to
0.degree. C. The suspension was filtered to provide ER-817664 as a
white crystalline solid.
Example 4b
Example 4b provides an alternate method of preparing compounds of
formula F-2 using the general scheme set forth at Schemes Vb and Vc
above. This method uses ER-817664 as an intermediate as prepared
according to Example 4a, above.
##STR00114##
ER-817664 (1 wt) was dissolved in ACN (10 v), the suspension was
cooled to 0.degree. C. and 2-acetoxy-2-methylpropanyl bromide (4.0
eq, 2.4 v) was added followed by the addition of H.sub.2O (1.0 eq.,
0.07 v). The resulting mixture was stirred at 0.degree. C. for 2
hours. NaHCO.sub.3 (sat. aqueous, 8.0 eq. 40 v) was added slowly at
0.degree. C. The resulting mixture was stirred at room temperature
for 30 minutes prior to extraction with MTBE (2.times.20 v). The
organic layer was washed with brine (5 v) and concentrated to give
the product as colorless oil.
##STR00115##
The starting bromide (1 wt), depicted immediately above, was
dissolved in toluene (10 v). DBU (1.8 eq., 0.73 v) was added and
the mixture was heated at 80.degree. C. The mixture was cooled to
room temperature, diluted with MTBE (20 v), and washed with water
(5 v) and then brine (5 v). The organic layer was then concentrated
to give the product as an off-white powder.
##STR00116##
The starting olefin compound (1 wt), depicted immediately above,
was dissolved in CH.sub.2Cl.sub.2 (5 v) and MeOH (5 v), and cooled
to between -40.degree. C. to -45.degree. C. The solution was then
treated with O.sub.3. Excess O.sub.3 was removed by N.sub.2 purge
and the solution was warmed to -15.degree. C. NaBH.sub.4 (1.0 eq,
0.18 wt) was added and the mixture was .[.warm.]. .Iadd.warmed
.Iaddend.to 0.degree. C. K.sub.2CO.sub.3 (1.3 eq.) was added and
the suspension stirred at rt. The reaction was neutralized with 1N
HCl (.about.4 eq, .about.20 v) at 0.degree. C. and the solution was
extracted with MTBE(10 v) to remove lypophilics. The aqueous layer
was concentrated to remove CH.sub.2Cl.sub.2 and MeOH. THF (4 v) was
added followed by NaIO.sub.4 (2 eq, 2 wt). The reaction was
extracted with MTBE (10 v) and n-BuOH (10 v). The combined organic
extracts were concentrated and the resulting powder was triturated
with EtOAc. After filtration the lactol was isolated as a pale
yellow powder.
##STR00117##
ER-818638 (1 wt) and LiCl (2.0 eq, 0.35 wt) was stirred in ACN (8.7
v). Hunig's base (1.5 eq) was added at 25.degree. C. 1 N HCl (5 v)
was added and the mixture was extracted with MTBE (10 v). The
organics were concentrated to provide ER-818640 which was used as
is in the next step.
##STR00118##
The starting .alpha.-olefin ester compound (1 wt), depicted
immediately above, was dissolved in MeOH (10 v) and added to 10 wt
% of Pd(C) (0.09 eq, .about.0.33 wt) under N.sub.2. The suspension
was then stirred under H.sub.2. The suspension was filtered through
a Celite.RTM. pad (20 wt), rinsing the filter cake with MeOH (20
v). The filtrate was concentrated and purified by flash
chromatography to give product as colorless oil (94.3% yield).
##STR00119##
Pyridine (10 eq.), Ph.sub.3P (7 eq.) and NIS (4 eq.) were added to
the solution of the ester (1 wt) in THF (15 v) separately. The
reaction mixture was stirred at ambient temperature. Aqueous citric
acid (20 wt %, 10 eq) was added and the mixture diluted with TBME
(30 v). The aqueous layer was separated and the organic layer
washed with water (5 v), aqueous Na.sub.2S.sub.2O.sub.3 (10% wt/v,
(5 v), water (5 v) and brine (5 v). The organic layer was
concentrated and purified by flash chromatography to give product
as colorless oil.
##STR00120##
The starting iodide (1 wt) was dissolved in MeOH (30 v) and heated
to 55.degree. C. NaBH.sub.4 (47 eq.) was added in 6 portions at
55.degree. C. over 80 minutes. The reaction was cooled to 0.degree.
C. and quenched with 1N HCl (30 v). After stirring 5 minutes, the
mixture was diluted with brine (30 v) and extracted with DCM (50
v.times.2). The organic layer was dried over Na.sub.2SO.sub.4 and
concentrated. The crude product was used directly in the next
step.
##STR00121##
The starting alcohol (1 wt), depicted immediately above, was
dissolved in EtOH (70 v) and Zn (165 eq.) was added. The suspension
was refluxed at 75-80.degree. C. The reaction mixture was cooled to
ambient temperature and 1N HCl (70 v) was added. The mixture was
extracted with DCM (3.times.100 v), the organic layer washed with
brine and concentrated.
##STR00122##
The starting lactone, as depicted immediately above, was dissolved
in DCM (50 v), Et.sub.3N (5.0 eq.), DMAP (0.3 eq.) and TBDPSCl (1.5
eq.) were added separately at ambient temperature under N.sub.2,
and the resulting solution was stirred at ambient temperature for
2-3 hours. Upon the completion of the reaction, the mixture was
diluted with TBME (100 v), washed with sat. aq. NaHCO.sub.3
solution (10 v), H.sub.2O (10 v) and brine (10 v). The organic
layer was concentrated and purified by flash chromatography to give
the product as colorless oil.
Example 4c
Example 4c provides another alternate method of preparing compounds
of formula F-2 using the general scheme set forth at Scheme VII
above. This method uses ER-811510 as an intermediate as prepared
according to Example 3, above where acetone is used instead of
cyclohexanone.
##STR00123##
ER-811510 (1 wt, 1 eq) was dissolved in methylene chloride (6.3 v)
and cooled to -5.degree. C. Pyridine (0.41 vol, 1.1 eq) was added
followed by bromoacetyl bromide (0.44 vol, 1.1 eq) while keeping
the temperature below 0.degree. C. The reaction was stirred at
.[.I.]. .Iadd.1 .Iaddend.hour and warmed to room temperature. Water
(8 vol) was added and the layers separated. The organic layer was
washed sequentially with aqueous copper sulfate pentahydrate (1.0
M, 10 vol), water (8 vol), and brine (10 vol) then dried over
magnesium sulfate, filtered and concentrated in vacuo to afford
ER-812771 as a tan solid.
##STR00124##
ER-812771 (1 wt, 1 eq) was dissolved in acetonitrile (6 v) and
triphenylphosphine was added and the reaction heated at 50.degree.
C. for 45 minutes. The reaction was cooled to -10.degree. C. then
1,8-diazabicyclo[5.4.0]undec-7-ene (DBU, 0.35 vol, 0.8 eq) was
added. The reaction was stirred for 15 minutes, heated to
80.degree. C. for 45 minutes then cooled to ambient temperature.
Ammonium chloride (saturated aqueous, 10 vol) was added and the
aqueous layer extracted with ethyl acetate (3.times.10 v). The
combined organic layers were dried over magnesium sulfate and
concentrated in vacuo. The crude product was purified by
chromatography to afford ER-812772 as a white solid.
##STR00125##
ER-812772 (1 wt, 1 eq) was dissolved in ethyl acetate (8 v). 10%
Palladium on carbon (0.05 wt, 0.01 eq) was added, the reaction
purged with nitrogen then stirred under hydrogen atmosphere for 2
hours. The catalyst was removed by filtration through Celite with
ethyl acetate washed. The combined filtrates were concentrated in
vacuo to afford ER-812829 as a white solid.
Example 5
Preparation of F-3a
##STR00126##
D-Gulonolactone (1 wt., 1 eq.), cyclohexanone (2 to 3 eq.), toluene
(6 vol.), and p-toluenesulfonic acid (0.021 wt., 0.02 eq.) were
charged to the reaction vessel. Reaction mixture was heated to
reflux with stirring. Upon azeotropic removal of water the reaction
was complete. The reaction mixture was cooled to 85 to 90.degree.
C. and agitation was increased. Heptane (5.2 vol.) was added over
20-30 minutes with stirring. Cooled to 65-70.degree. C. and stirred
for 30 minutes at 65-70.degree. C. The solid product was filtered
at 65-70.degree. C., maintaining mother liquor temp .[.35.degree.
C.]. .Iadd.>35.degree. C.Iaddend.. Re-filtered at 35-40.degree.
C. and maintained the mother liquor at ambient temperature for 30
minutes. Re-filtered the mother liquor. The filter cake was washed
two times with heptane (2.times.1.7 vol.) then dried to afford
ER-805715. Yield 84% (1.6 wt.).
In an alternate method for preparing ER-805715, D-gulonolactone (1
wt), cyclohexanone (1.32 wt, 2.4 eq), p-TsOH-monohydrate (0.02 wt,
0.02 eq) and toluene (12 vol) were refluxed together for 19 hours,
while azeotropically removing water. The mixture was washed with 5%
aqueous NaHCO.sub.3 (4 vol) followed by saturated aqueous NaCl (2
vol.times.2, pH=7). The organic phase was concentrated by
distillation (ca. 4.5 vol of toluene remaining) and cooled to
100.degree. C. before heptane (10 vol) was added, maintaining
internal temperature .[.80.degree. C.]. .Iadd.>80.degree.
C.Iaddend.. The mixture was heated to reflux for at least 1 hour
before it was cooled to and aged at 85.degree. C. for 3 hours, at
80.degree. C. for 3 hrs and then cooled to 40.degree. C. in 12 hrs.
The product was collected by filtration and the cake washed with
heptane (2 vol). The filter cake was dried by airflow to afford
ER-805715 (1.48 wt) in 78% of yield.
##STR00127##
ER-805715 (1 wt., 1 eq.) was charged to reaction vessel and
dissolved in anhydrous THF (3.34 vol.) and anhydrous toluene (2.5
vol.). The mixture was cooled to -15 to -10.degree. C. DIBALH (1.5M
in toluene, 2.4 vol., 1.2 eq.) was added over 1 hour and the
mixture stirred for 15-30 minutes at -15 to -10.degree. C. The
reaction was inverse quenched into KNa-Tartrate solution (1 wt. KNa
Tartrate in 2.9 wt. water) at 10.degree. C. and the resulting
mixture allowed to warm to room temperature and stir for 4 hours.
The mixture was filtered then the layers separated and extracted
with MTBE (2 vol.). The organic layers were combined and the
solvents removed in vacuo to afford ER-805814. Yield 100%, (1.02
wt.).
##STR00128##
ER 805814 (1 wt.) was dissolved in anhydrous THF (3.3 vol.) and
treated with (methoxymethyl)triphenylphosphonium chloride (2.11
wt., 2.1 eq.). The reaction mixture was heated to 28-32.degree. C.
then a solution of KOtBu (0.66 wt., 2 eq.) in anhydrous THF (2.64
vol.) was added over 100-140 minutes, maintaining reaction
temperature 30-35.degree. C. After 5 hours, the reaction was cooled
to 20-25.degree. C., MTBE (5.11 vol.) was added and the mixture
stirred. Brine (3 wt.) and water (3 wt.) were added (exothermic at
start of addition, controlled by bath @20-25.degree. C.). Organic
layer was separated and treated with a solution of maleic anhydride
(0.27 wt.) in MTBE/THF (1/1 v/v, 1.78 vol.). NaOH solution (0.088
wt. in 2.5 vol. water) was added slowly to the reaction mixture.
The organic layer was concentrated to give crude ER 805815 (0.985
wt.). The residue was triturated three times with MTBE/heptane (1/4
v/v, 6.6 vol.). The extract was filtered through SiO.sub.2 (3 wt),
eluting with MTBE/heptane (1/2 v/v, 45 vol.). The filtrate was
concentrated to give ER-805815 (0.88 wt., 81% yield).
In an alternate method for preparing ER-805815, a solution of
t-BuOK (0.989 wt, 3 eq) in THF (4 wt) was added to a suspension of
(methoxymethyl)triphenylphosphonium chloride (3.12 wt, 3.1 eq) in
THF (1.78 wt), maintaining the reaction temperature between
0-10.degree. C. The addition vessel was rinsed with THF
(2.times.0.7 wt). A solution of ER-805814 (1 wt, 1 eq) in THF (1.42
wt) was added to the reaction, maintaining 0-10.degree. C. The
addition vessel was rinsed with THF (2.times.0.7 wt). The mixture
was stirred at 20-30.degree. C. overnight and 30-35.degree. C. for
3 hours. The reaction was cooled below 30.degree. C. and diluted
with MTBE (3.7 wt) followed by 10 wt % aqueous NaCl (4 wt)
solution. The mixture was stirred for 30 minutes and the layers
were separated. Maleic anhydride (0.63 wt, 2.2 eq) was added and
the mixture stirred at room temperature for 30 minutes. Water (6
wt) and a solution of NaOH (48 wt %, 0.64 wt, 2.6 eq) was added
dropwise, maintaining the reaction below 15.degree. C. After
stirring below 15.degree. C., the lower layer was separated. Water
(6 wt) was added followed by a solution of NaOH (48 wt %, 0.64 wt,
2.6 eq), keeping the mixture below 15.degree. C. during the
addition. After stirring below 15.degree. C., the lower layer was
separated. The organic layer was washed three times with a 15 wt %
aqueous NaCl solution (3.times.4 wt). The organic layer was
concentrated in vacuo. The residue was diluted with MTBE (1 wt) and
concentrated in vacuo. The residue was diluted dropwise with IPE (3
wt) at 40-50.degree. C. over 30 minutes. The suspension was stirred
for 1 hour at 40-50.degree. C. and slowly cooled to 0-10.degree. C.
and stirred for 1 hour. The solids were filtered and the cake
washed with IPE (2 wt). The filtrate and washings were concentrated
in vacuo. The residue was treated with MeOH (2.37 wt) and water
(0.4 wt) and extracted with heptane (2.74 wt). The lower layer was
extracted 9 times with heptane (2.05 wt). The extracted solutions
were combined and concentrated in vacuo to give ER-805815 (1.07 wt,
98.6%).
In an alternative method for workup of ER-805815, the crude organic
layer that is produced following brine wash and concentration is
treated with MTBE (2.86 wt) and celite (0.5 wt). After stirring for
2.5 h, heptane (1.46 wt) was added over 2 hrs and the mixture
stirred overnight. The precipitate was filtered. The filter cake
was washed with MTBE/Heptane (1:1) (5 wt). The filtrate was
concentrated in vacuo until the volume was decreased to about 3
volume. The residue was dissolved in MeOH (2 wts) and H.sub.2O (6
wts). The mixture was extracted with heptane/MTBE (5:1) (3*6 wts).
The organic layer was separated and concentrated to provide
ER-805815 which was used as is for the following step.
##STR00129##
ER-805815 (1 wt) was dissolved in acetone (2.4 vol) and water (0.4
vol). N-Methylmorpholine N-oxide (0.62 wt, 2 eq) was added and the
mixture cooled to 0-5.degree. C. OSO.sub.4 (0.15M in water, 0.065
vol) was added and the reaction was maintained at 0-5.degree. C.
The reaction mixture was stirred at 0-5.degree. C. for 12 hours.
Water (0.2 vol) was added over 1 hour at 0-2.degree. C. The mixture
was stirred for one hour at 0-5.degree. C. The product was filtered
and the solids washed twice with pre-cooled (0-5.degree. C.)
acetone/water (1/1, v/v, 2.times.0.7 vol). The product was dried to
afford ER-805816 (0.526 wt, 52% yield, residual Os <17 ppm).
In an alternate method for preparing ER-805816, a solution of
ER-805815 (1 wt, 1 eq) in acetone (4 wt) was charged into a
four-necked flask, then water (0.5 wt) was added at ambient
temperature. To the mixture was added anhydrous
N-methylmorpholine-N-oxide (0.38 wt, 1.2 eq). Potassium osmate
dihydrate (0.003 wt, 0.003 eq) was added portion-wise at 25 to
35.degree. C. while cooling with water. The mixture was kept at
this temperature for 4 hours. A solution of sodium thio sulfate
0.075 wt, 0.49 eq) in water (0.5 wt) was added at ambient
temperature, then the mixture was stirred for 0.5 hour. The mixture
was cooled to 0-5.degree. C. and stirred for 2 hours. The resulting
precipitate was collected and the wet cake was washed with methanol
(0.6 wt) and water (1.5 wt) to obtain the crude product (1.25 wt).
The crude product sample was dried (0.611 wt). The crude ER-805816
(1.25 wt) was added to water (3.05 wt) and stirred for 2 hours at
about 25.degree. C. The precipitate was filtered and washed with
water (1.53 wt) to afford the crude wet cake (1.05 wt). The crude
product sample was dried and sampled, ICP Os=37 ppm. The crude
ER-805816 (1.05 wt) was added to water (2.81 wt) and stirred for 2
hours at about 25.degree. C. The precipitate was filtered and
washed with water (1.4 wt) and methanol (0.45 wt) to afford the
crude ER-805816 (0.736 wt). The wet cake was dried crude product
(0.56 wt, ICP (Os)=28 ppm). ER-805816 (0.56 wt) was dissolved in
acetone (1.76 wt) at 45 to 55.degree. C. To the solution was added
active carbon (0.027 wt) and stirred at same temperature for 0.5
hour. The mixture was filtered and the cake was washed with hot
acetone (0.214 wt). The filtrate was kept at 45 to 50.degree. C.
and water (0.83 wt) was added over 10 minutes and temperature was
kept at 40 to 50.degree. C. during water addition. The mixture was
cooled to 0 to 5.degree. C. and stirred for 1.5 hours. The white
precipitate was filtered and washed with a solution of acetone
(0.17 wt) and water (0.22 wt) then dried to give ER-805816 (0.508
wt, 0.49 eq, KF 5.0%, ICP (Os) 9.6 ppm).
##STR00130##
ER-805816 (1 wt) was slurried in acetic acid (0.89 vol, 5.8 eq) and
acetic anhydride (3.57 wt, 13 eq). Anhydrous ZnCl.sub.2 (0.2 wt,
0.54 eq) was added. Reaction mixture was stirred for 24 hours
18-22.degree. C. Reaction was quenched into ice (5 wt) and water (5
vol). EtOAc (10 vol) was added with stirring and the aqueous layer
is separated. The aqueous layer was back extracted with EtOAc (10
vol). The combined organic layers were washed sequentially with
brine (10 vol), 5% aqueous NaOAc (6 vol), and brine (6 vol). The
organic layer was concentrated. The crude concentrate was dissolved
in 25% EtOAc/hex (4 vol) and filtered through SiO.sub.2. The pad
was washed with 25% EtOAc/hex (2.times.12 vol) and further 25%
EtOAc/hex (48 vol). The organic layer was concentrated to give
ER-805819 (1 wt, 81%).
In an alternate method for preparing ER-805819, zinc chloride (0.2
wt, 0.54 eq), acetic anhydride (2.75 wt, 10 eq), and acetic acid (1
wt, 6 eq) were combined. The mixture was cooled to 15-20.degree. C.
ER-805816 (1 wt, 1 eq) was added, maintaining the internal
temperature at 15 to 30.degree. C. The mixture was then stirred at
35-40.degree. C. for 6 hours. The reaction mixture was cooled below
25.degree. C. Methanol (3.2 wt, 4 vol) was added drop-wise
maintaining reaction temperature below 25.degree. C. Heptane (2.7
wt, 4 vol) was added. Water was added (4 wt, 4 vol) maintaining
reaction temperature below 25.degree. C. The mixture was stirred
for 15 minutes, and then the phases were separated. The lower layer
was washed twice with heptane (2.7 wt, 4 vol) and the heptane
layers were discarded. The lower layer was extracted twice with
toluene (6.1 wt, 8 vol). The combined toluene layers were washed
twice with 17 wt % potassium bicarbonate aqueous solution (0.82 wt
KHCO3 in 3.98 wt water, 4.36 vol), twice with water (4 wt), and
concentrated. Methanol (3.95 wt, 5 vol) was added at 25-30.degree.
C. and the mixture stirred for 10 minutes. Water (0.3 wt) was added
at 25-30.degree. C. The mixture was cooled to 0.degree. C. and
seeded. The mixture was stirred at 0.degree. C. for 1 hour. Water
(0.7 wt) was added drop-wise over 1 hour. Water (4 wt) was added
drop-wise over 1 hour. The resulting precipitated solids were
filtered, and the filter cake washed twice with a 0.degree. C.
methanol (1.03 wt) and water (0.7 wt) solution. The cake was dried
to afford ER-805819 (0.99 wt, 0.84 eq).
##STR00131##
ER-805819 (1 wt) was dissolved in anhydrous acetonitrile (15 vol)
and treated with methyl 3-trimethylsilylpent-4-eneoate (0.93 vol, 2
eq). The reaction mixture was cooled to 0-5.degree. C. and
BF.sub.3OEt.sub.2 (0.54 vol, 1.95 eq) was added over 5 minutes,
maintaining reaction temperature between 0-5.degree. C. Reaction
mixture was stirred 0-5.degree. C. for 12 hours. Reaction was
quenched into saturated sodium bicarbonate (20 vol) with vigorous
stirring. Extracted twice with EtOAc (2.times.8 vol). The combined
organics were washed with brine (12 vol) and concentrated to give
ER-805821 (1 wt, 88% yield, use as is).
In an alternate method for preparing ER-805821, ER 805819 (1 wt, 1
eq) and .[.ER.]. methyl 3-trimethysilylpent-4-eneoate (0.93 vol, 2
eq) were dissolved in anhydrous acetonitrile (5.46 wt, 7 vol). The
reaction mixture was cooled to 0-5.degree. C. and BF.sub.3.[..].
.Iadd.-.Iaddend.OEt.sub.2 (0.54 vol, 1.95 eq) was added over 5
minutes, while maintaining reaction temperature between 0-5.degree.
C. The reaction mixture was stirred at 0-5.degree. C. for 20 hours
then heptane (5.47 wt, 8 vol) was added at 0-5.degree. C. The
phases were separated and the lower layer treated with heptane
(5.47 wt, 8 vol) at 0-5.degree. C. The reaction was quenched by
dropwise addition of 7.4% potassium bicarbonate aqueous solution
(0.64 wt KHCO.sub.3 and 8 wt water), while maintaining the reaction
temperature at 0-15.degree. C. Toluene (8.65 wt, 10 vol) was added
and the mixture stirred for 30 minutes. The lower layer was
separated and the upper organic layer washed twice with water (10
vol) and concentrated to afford ER-805821 as a crude oil (1.05 wt,
0.935 eq).
##STR00132##
ER-805821 (1 wt) was dissolved in anhydrous THF (8.4 vol) and
anhydrous MeOAc (2 vol). Triton B(OH) (3.6 vol) was added over 2
minutes, reaction maintained 17-23.degree. C. Reaction was stirred
for 1.5 hour. Reaction mixture was filtered. The filtrate was
concentrated and passed through a pad of SiO.sub.2 (5 wt, EtOAc, 20
vol). The filtrate was washed with brine (2.2 vol) and evaporated
to give ER-805822 (0.54 wt, 72% yield).
In an alternate method for preparing ER-805822, ER-805821 (1 wt, 1
eq, 11.18 g, 21.81 mmol) was dissolved in anhydrous MTBE (4.4 wt, 6
vol.) and cooled to 0-5.degree. C. NaOMe (28 wt % in MeOH, 0.564
wt, 1.5 eq) was added to the mixture over 1 hour at 0-5.degree. C.
and stirred for 3 hour at same temperature range. The reaction was
quenched by addition of acetic acid (0.188 wt, 1.6 eq.),
maintaining 0-5.degree. C. during addition. The mixture was stirred
overnight then treated with a 5 wt % aqueous solution of KHCO.sub.3
(3 wt) and ethyl acetate (3.6 wt, 4 vol) at 0-5.degree. C., and
then stirred for 15 minutes. After phase separation, the lower
layer was extracted with ethyl acetate (3.6 wt, 4 vol) twice. The
combined organic layer was concentrated. To the residue was added
acetone (2 wt, 2.5 vol) and IPE (2 wt, 2.7 vol) and stirred
overnight at 0-5.degree. C. The mixture was filtered through
Celite.RTM. (0.25 wt) and washed with acetone (2 wt). The filtrate
was concentrated to afford the crude oil (0.55 wt). To the residue
was added acetone (0.2 wt, 0.25 vol.) and IPE (0.54 wt, 0.75 vol.)
and stirred for 1 hour at 40-50.degree. C. The solution was seeded
with ER-805822 at room temperature and stirred overnight at room
temperature. To the suspension was added IPE (1.27 wt, 1.75 vol.)
over 2 hours at room temperature. After stirring for 5 hours at
room temperature, the precipitate was collected by filtration, and
the cake washed with acetone/IPE (1/10) (2 vol.). The obtained cake
was dried in a tray-type chamber at 30-40.degree. C. overnight to
afford the desired product ER-805822 (0.286 wt, 0.38 eq) in 38.0%
yield from ER-805819.
In an alternative method for workup of crude ER-805822, the residue
following concentration of the final EtOAc solution was dissolved
in IPA (2 wt) and the solution was heated to 50.degree. C. Heptane
(5 wts) was added and the mixture was cooled to 20.degree. C. and
seeded. The mixture was stirred at 20.degree. C. overnight. Heptane
(10 wts) was added and the mixture was cooled to -5.degree. C. in
30 min and stirred at -5.degree. C. for 5 hrs. The mixture was
filtered and the filter cake was washed with heptane (2 wts). The
filter cake was dried with air flow under vacuum to provide
ER-805822 (60%).
##STR00133##
ER 805822 (1 wt) was dissolved in ethyl acetate or another
appropriate solvent (5 vol) and water (5 vol). NaIO.sub.4 (0.58 wt,
1.05 eq) is added portionwise over 30 min to 1 hour, maintaining
reaction temperature 0-10.degree. C. Reaction is stirred for up to
2 hours. The reaction mixture was treated with NaCl (1 wt) and
stirred for 30 min at 0 to 10.degree. C. The reaction mixture was
filtered and the cake is rinsed with ethyl acetate (2 vol). The
phases were separated and the lower layer extracted with EtOAc (5
vol) three times. The combined organic layer was washed with 20%
aqueous NaCl (5 wt). The organic layer was concentrated to give ER
804697 (1 wt). The residue was dissolved in toluene (2 vol) and the
solution concentrated. The residue was dissolved in acetonitrile (7
vol) and used for the next step.
##STR00134##
NiCl.sub.2 (0.025 wt) and CrCl.sub.2 (2.5 wt) were charged to
reaction vessel under inert atmosphere. Anhydrous dichloromethane
(5 vol) was charged. Stirring was initiated and the mixture was
cooled to 0-3.degree. C. Anhydrous DMSO (6.7 vol) was added with
vigorous stirring over 45 minutes, maintaining temperature below
20.degree. C. ER-804697 (1 wt) was dissolved in anhydrous
dichloromethane (1 vol) and charged to the reaction vessel. The
resulting mixture was warmed to 25.degree. C. and
1-bromo-2-trimethylsilylethylene (2.58 wt) was added neat over 20
minutes. The reaction temperature was maintained below 45.degree.
C. The reaction was stirred for 30 minutes at 25-35.degree. C.
following complete addition. Methanol (5 vol) was added and the
mixture was stirred for 10 minutes. MTBE (33 vol) was charged and
the slurry transferred into 1N HCl (25 vol) and water (10 vol). The
mixture was stirred for 5 minutes. The aqueous layer was back
extracted with MTBE (10 vol) and the combined organics washed
sequentially with 0.2N HCl (17 vol), twice with 1% NaCl solution
(2.times.17 vol), and brine (13 vol). The organic layer was
concentrated and purified (SiO.sub.2, 25 wt, 10 column vol
EtOAc/Hex 1/3.5 v/v) to give ER-804698 (0.53 wt, 61%).
In an alternate method, this reaction was performed in the presence
of the chiral ligand ER-807363 in a manner substantially similar to
that described for the preparation of ER-118047, infra.
In an alternate method for preparing ER-804698, DMSO (7 vol.) and
MeCN (7 vol) were degassed and cooled to 0-10.degree. C. The
solution was treated portionwise with CrCl.sub.2 (10 eq, 3.47 wt)
and NiCl.sub.2 (0.1 eq, 0.037 wt) such that the internal
temperature did not exceed 20.degree. C. A solution of ER-804697 (1
wt, 1 eq) in MeCN (7 vol) and 1-bromo-2-trimethylsilylethylene (5
eq, 2.5 wt) were added dropwise at 0-10.degree. C., not allowing
the internal temperature to exceed 15.degree. C. The reaction
mixture was stirred at 5-15.degree. C. overnight. To the mixture
was added methanol (5.5 wt), water (7 wts), and MTBE (5.2 wts). The
reaction was stirred for 1 hour and the lower layer was separated
(layer 1). To the upper layer was added a premixed solution of NaCl
(1.5 wts) and water (13.5 wts). The mixture was stirred for 1 hour
and the lower layer was separated (layer 2). To the upper layer was
added heptane (4.8 wts), methanol (2.8 wts), and a premixed
solution of NaCl (1.5 wts) and water (13.5 wts). The mixture was
stirred for 1 hour and the lower layer was separated (layer 3). The
upper layer was drained and saved (organic 1). The reactor was
charged with layer 1, methanol (2.8 wts), and MTBE (2.8 wts). The
mixture was stirred overnight. The lower layer was separated and
discarded. The upper layer was treated with layer 2. The mixture
was stirred for 1 hour and the lower layer was separated and
discarded. The upper layer was treated with layer 3 and heptane
(4.8 wts). The mixture was stirred for 1 hour and the lower layer
was separated and discarded. The upper layer was drained and saved
(organic 2). The reactor was charged with layer 3, MTBE (0.8 wts),
and heptane (2.7 wts). The mixture was stirred for 1 hour and the
lower layers was separated and discarded. The upper layer was
combined with organic 1 and organic 2. The combined organics were
filtered and concentrated at reduced pressure to afford the crude
ER-804698 which was purified by chromatography (SiO.sub.2, 25 wt,
10 column vol EtOAc/Hex 1/3.5 v/v) to give ER-804698 (0.67 wt, 57%
yield).
In an alternate method of preparation of ER-804698, the crude
material is taken directly to the next step without
purification.
##STR00135##
ER 804698 (1 wt, 1 eq) was treated with AcOH (4.2 wts) and water
(4.2 wts). The mixture was heated to 90-97.degree. C. for 100 min.
The mixture was cooled to below 15.degree. C. then washed with
heptane (2.times.2.7 wts) twice below 15.degree. C. After phase
separation, a mixture of 20 wt % aqueous KHCO.sub.3 solution (7.7
wts, 35 eq) and MTBE (5.95 wts) was added dropwise to the lower
layer such that temperature does not exceed 15.degree. C. After
phase separation the upper layer was washed successively with 5 wt
% aqueous KHCO.sub.3 solution (0.2 wts), and twice with 5 wt %
aqueous NaCl solution (2.times.0.2 wts). The organic layer was
concentrated under reduced pressure and MTBE (1.49 wts) was added.
The mixture was heated to 55.degree. C. and stirred until
dissolved. Heptane (1.00 wts) was added to the solution and the
solution was cooled to 40-45.degree. C. Additional heptane (4.47
wts) was added to the solution and the solution was cooled to
5-15.degree. C. and then stirred overnight. The crystals were
filtered and rinsed with heptane to provide ER-807023 (0.58 wts,
71% yield).
##STR00136##
ER-807023 (1 wt, 1 eq) and MTBE (7.43 wts) were charged to a
reactor under a nitrogen atmosphere. To the reaction was added
2,6-lutidine (2.15 wts, 7.5 eq). To the mixture was added dropwise
TBSOTf (2.47 wts, 3.5 eq) at 0.degree. C. The reaction mixture was
stirred for 30 min at 0-10.degree. C., then warmed to 23.degree. C.
over 1 hr and held at 23.degree. C. for 16 hrs. MeOH (0.21 wts, 2.5
eq.) and water (14.8 wts) were added dropwise sequentially to the
reaction mixture, maintaining temperature below 30.degree. C. After
phase separation, the upper layer was washed with 1N aqueous
hydrochloric acid (16.2 wts), 5% NaCl aq. (14.8 wts), 5%
NaHCO.sub.3 aq. (14.8 wts), 5% NaCl aq. (14.8 wts), and 5% NaCl aq.
(14.8 wts), respectively. The upper organic layer was concentrated
by distillation under reduced pressure to afford the crude
ER-804699. MeOH (7.91 wts) was added and the mixture was heated to
50.degree. C. for 30 min. The mixture was cooled to 0.degree. C.
over 5 h, and then stirred overnight at 0.degree. C. The solid was
filtered, and the cake was washed with cold MeOH (4 wts) and dried
to yield ER 804699 (1.42 wts, 74% yield).
##STR00137##
Into a reactor under a nitrogen atmosphere was charged a solution
of ER-804699 (1 wt, 1 eq) in toluene (2.60 wts). Acetonitrile (4.72
wts) was added. TBSCl (0.011 wts, 0.05 eq) was added. The reaction
mixture was warmed to 30.degree. C. and the NIS was added (1.25
wts, 4 eq). The reaction mixture was stirred at 22 hrs at
30.degree. C. The reaction was cooled to 25.degree. C. and the
mixture of aqueous sodium thiosulfate and sodium bicarbonate (10.35
wts) were added over 10 minutes keeping the internal temperature
below 30.degree. C. The reaction was stirred for 30 minutes at
25.degree. C. The aqueous layer was separated. The upper layer was
washed twice with 10% NaCl (aq) (2.times.9.9 wts). The organic
layer was concentrated under reduced pressure to give crude
ER-803895 that was purified using silica gel chromatography to
provide ER-803895 (0.96 wt, 89.5% yield).
Example 6
Assembly of F-1a, F-2a, and F-3a and Preparation of B-1939
A. Preparation of (R) or (S)
N-[2-(4-Isopropyl-4,5-dihydro-oxazol-2-yl)-6-methyl-phenyl]-methanesulfon-
amide
##STR00138##
A pre-dried glass lined reactor was charged with triphosgene (1
wt., 1 eq.) and anhydrous THF (2 vol.) and was cooled to an
internal temperature of -10.degree. C. A second pre-dried glass
lined .Iadd.reactor .Iaddend.was charged with ER-807244 (1.27 wt.,
2.5 eq.) and anhydrous THF (3 vol.) then cooled to an internal
temperature of -10.degree. C. The contents of the first reactor
were transferred into the second reactor at a rate such that
internal temperature did not exceed 15.degree. C. After complete
addition, the reaction was stirred at an internal temperature of
0.degree. C. for 1 hour and then gradually warmed to 25.degree. C.
A sparge of nitrogen was used for 18 hours to scrub away excess
phosgene with trapping of the off-gases through a 2 N NaOH
solution. MTBE (3 vol.) was added and the solvent removed by
distillation under N.sub.2 purge at 40.degree. to 46.degree. C.,
adding more MTBE as needed. Upon complete removal of the phosgene,
the mixture was cooled to an internal temperature of 5.degree. to
10.degree. C. and the solution filtered with MTBE (3 vol.) washes
to yield ER-807245 (1.12 wt., 0.97 eq.) as a white crystalline
solid.
##STR00139##
Into a pre-dried and inerted reactor 1, was added ER-807245 (1 wt.,
1 eq.) and anhydrous DMF (4 vol.). With stirring, the mixture was
heated to an internal temperature of 95.degree. C. D or L-Valinol
(1.05 eq., 0.61 wt.) was dissolved in anhydrous (DMF 1.3 vol.) in
reactor 2 with heating to an internal temperature of 90.degree. C.
The contents of reactor 2 were transferred into reactor 1 at
internal temperature 90.degree. C. CO.sub.2 evolution was .[.be.].
observed and the reaction was vented with a N.sub.2 bleed. The
reaction solution was stirred at 90.degree. C. for 3 hours and then
cooled to an internal temperature of 65.degree. C. Then, an aqueous
slurry of lithium hydroxide (0.47 wt., 2 eq.) in water (2 vol.) was
added to reactor 1 and the suspension stirred at an internal
temperature of 65.degree. C. for 1 hour. The reactor was charged
with water (5 vol.) cooled to an internal temperature of
.about.5.degree. C. over 3 hours. The mixture was stirred for 8
hours at internal temperature .about.5.degree. C. and the desired
product collected by filtration with water (2.times.4 vol.) washes
followed by n-heptane (2.times.3 vol.). The product was dried under
vacuum and N.sub.2 flow at 35.degree. C. for 24 hours or until
KF.ltoreq.250 ppm to yield ER-806628 or ER-808056 (0.80 wt., 0.60
eq.) as a crystalline solid.
##STR00140##
A pre-dried and inerted reactor under nitrogen was charged with
ER-806628 or ER-808056 (1 wt., 1 eq.), pyridine (3 wt., 11.4 eq.)
and DMAP (0.03 wt., 0.05 eq.). The reaction was cooled to an
internal temperature of -10.degree. C. then methanesulfonyl
chloride (1.46 wt., 3 eq.) was added at a rate such that internal
temperature was below 15.degree. C. Upon complete addition, the
reaction was stirred at an internal temperature of 0-15.degree. C.
for 1 hour and then slowly warmed to 25.degree. C. over 2 hours.
MTBE (2.6 vol.), was added followed by process water (2 vol.) at a
rate such that the internal temperature did not exceed 35.degree.
C. The biphasic mixture was titrated with 6N hydrochloric acid,
(.about.1.9 vol.) portion-wise until the pH of the aqueous
layer=.about.3 to 5. If pH went under 3, 30% (w/w) aqueous solution
of Na.sub.2CO.sub.3 was added to back titrate to the desired pH.
The phases were allowed to partition and the aqueous phase
separated. All organics were combined with water (0.7 vol.) and the
aqueous phase discarded. The MTBE was distilled to a level of
.about.2 vol. at atmosphere pressure to constant bp 55.degree. C.
and KF <500 ppm. Additional MTBE was added if necessary. The
solution was cooled to an internal temperature of 5-10.degree. C.
with seeding when necessary to induce crystallization. n-Heptane
(0.5 vol.) was added and the mixture stirred for 18 hours at
5.degree. C. ER-806629 or ER-807363 was collected by filtration
with n-heptane (2.times.3 vol.) washes. A second crop of crystals
was obtained by concentration of the filtrates to 1/2 volume and
cooling to 0.degree. C. The filter cake was dried under N.sub.2 for
18 hours. The crude weighted ER-806629 was charged into a pre-dried
reactor and MTBE (3 vol.) was added. The resulting mixture was
heated to an internal temperature of 45-50.degree. C. for 45
minutes and then slowly cooled to 5.degree. C. over 3 hours, with
seeding when necessary. n-Heptane (0.5 vol.) was added and the
mixture stirred for 18 hours at an internal temperature of
5.degree. C. The solid product was collected via filtration and
n-heptane (2.times.3 vol.) washes then dried under vacuum at
35.degree. C. for 24 hours to yield ER-806629 or ER-807363 (1.7
wt., 0.57 eq.) as a crystalline solid.
B. Assembly of F-1a and F-2a Intramolecular Ether Formation
##STR00141##
An appropriately sized reactor 1 was charged with ER-807363 (1.82
wt, 3.55 eq) and the atmosphere was exchanged for nitrogen.
Anhydrous THF (15 vol) was added. In reactor 2, ER-806067 (F-1a,
1.14 wt, 1.1 eq) and ER-805973 (F-2a, 1 wt, 1 eq) were combined and
dissolved in anhydrous THF (6.3 vol). With stirring, both reactors
were sparged with nitrogen for 30-45 minutes. Under an inert
atmosphere, reactor 2 was charged with CrCl.sub.2 (0.75 wt, 3.55
eq) and then heated to an internal temperature of 30.degree. C.
Reactor 2 was charged with triethylamine (0.62 wt, 3.55 eq) at a
rate such that internal temperature did not exceed 45.degree. C.
After complete addition, an internal temperature of 30.degree. C.
was maintained for 1 hour. After 1 hour, reactor 2 was cooled to
0.degree. C. and charged in an inert fashion with NiCl.sub.2 (0.02
wt, 0.1 eq), followed by the contents of reactor 1 and the reaction
was warmed to rt. Reactor 2 was cooled to an internal temperature
of 0.degree. C. and then ethylenediamine (1.2 vol, 10 eq) was added
at a rate such that the internal temperature did not exceed
10.degree. C. Note: An exotherm was observed. The reaction was
stirred for 1 hour, and then water (8 vol) and n-heptane (20 vol)
were added and the biphasic mixture stirred for 4 minutes and the
layers allowed to partition. The organic layer was separated and
the aqueous layer back extracted with MTBE (20 vol). The combined
organic layers were concentrated in vacuo to a crude oil followed
by an azeotrope with anhydrous THF (2.times.10.5 vol). The crude
product was dissolved in anhydrous THF (4.5 vol) and then stored at
-20.degree. C. until utilization in the next stage.
##STR00142##
The ER-808227/THF solution from the previous step was analyzed via
KF analysis. If KF <1000 ppm, then proceeded. If .[.KF 1000
ppm.]. .Iadd.KF>1000 ppm.Iaddend., azeotroped in vacuo with
anhydrous THF (4.1 vol.). Repeated azeotrope until specification
was met. The final solution meeting specifications contained the
dissolved crude ER-808227 in anhydrous THF (4.1 vol.). Once the
specification was met, an appropriately sized inerted reactor was
charged with anhydrous THF (106 vol.) and the ER-808227/THF
solution from the previous step. The reactor was cooled to an
internal temperature of -15 to -20.degree. C., then 0.5 M KHMDS in
toluene (9.1 wt., 3.0 eq.) was added at a rate such that internal
temperature did not exceed -12.degree. C. Approximately 4.5 eq.
KHMDS was necessary to drive the reaction to completion. The
reaction was reverse quenched into semi-saturated ammonium chloride
(40 vol.) at an internal temperature of 0.degree. C. n-Heptane (80
vol.) was added, stirred for 2-5 minutes, and then allowed to
partition. The organic layer was separated, the aqueous layer was
back extracted with MTBE (70 vol.), then the organic layers were
combined and washed with saturated sodium chloride solution (70
vol.). The organic layer was separated and concentrated in vacuo.
To the crude concentrate was added n-heptane (60 vol.). Note:
ER-807363 precipitated out of solution. The resulting suspension
was filtered and the solids washed with n-heptane (20 vol.). The
filtrate was concentrated in vacuo to afford crude ER-806746
(.about.4 wt.) as a brown oil.[.:.]..Iadd.. .Iaddend.Note: When
additional ER-807363 precipitated out of solution, the filtration
process was repeated. The crude ER-806746 was purified via
SiO.sub.2 column chromatography to yield ER-804027 (1.16 wt., 0.55
eq.) as a clear yellowish oil. The chromatography was performed as
follows: the column was first flushed with sufficient MTBE to
remove water then flushed with heptane to remove the MTBE. The
ER-806746 was loaded onto the column as a solution in heptane then
eluted from the column with heptane/MTBE (5:1) then heptane/MTBE
(4:1) with the fractions monitored at 230 nm by UV detector.
##STR00143##
A reactor was charged with ER-804027 (1 wt, 1 eq) and anhydrous
dichloromethane (7.6 vol). The reactor was cooled to an internal
temperature of -78.degree. C. and then 1 M DIBALH in
dichloromethane (3.0 wt, 2.25 eq) was added at a rate such that
internal temperature did not exceed -60.degree. C. Methanol (0.1
vol) was added at a rate such that internal temperature did not
exceed -60.degree. C. Note: hydrogen gas evolved and was diluted
with a stream of nitrogen. Upon complete addition, the mixture was
warmed to ambient temperature and then 1 N hydrochloric acid (10.6
vol) and MTBE (25 vol) were added. The mixture was stirred for 20
minutes and the layers allowed to partition. The organic layer was
separated and the aqueous layer .Iadd.was .Iaddend.back extracted
.[.layer.]. with MTBE (15.3 vol). The organic layers were combined
and washed with water (3 vol), saturated sodium bicarbonate (3
vol), and saturated sodium chloride (3 vol), respectively, then
concentrated in vacuo. The crude concentrate was purified via
SiO.sub.2 column chromatography to yield ER-804028 (0.84 wt, 0.93
eq) as a white foam.
C. Incorporation of F-3a and Transformations to B-1939
##STR00144##
ER-803895 (F-3a) was dissolved in anhydrous toluene (14 wt.) and
cooled to .[..rarw.75.degree. C..]. .Iadd.<-75.degree. C.
.Iaddend.under an argon atmosphere. DIBALH (1.5M in toluene, 0.95
wt., 1.3 eq.) was added at a rate to maintain the internal reaction
temperature .[..rarw.70.degree. C..]. .Iadd.<-70.degree. C.
.Iaddend.The resulting mixture was stirred for 30 minutes then
quenched with anhydrous methanol (0.13 wt., 3.2 eq.), maintaining
the internal reaction temperature .[..rarw.65.degree. C..].
.Iadd.<-65.degree. C. .Iaddend.The reaction mixture was allowed
to warm to -10.degree. C. and transferred with an MTBE rinse (3.74
wt.) to a workup vessel containing 1N HCl (10.2 wt.). The mixture
was stirred for 30 minutes and the aqueous layer .[.is.]. .Iadd.was
.Iaddend.drained. The organic phase was washed sequentially with 1N
HCl (10.2 wt.), water (10 wt.), saturated aqueous sodium
bicarbonate (10 wt.), and brine (10 wt.) then concentrated under
reduced pressure. The concentrate was purified via silica gel
chromatography to afford ER-803896 (0.96 wt., 93% yield). The
product is stored at -20.degree. C. under argon.
##STR00145##
At 0.degree. C., a solution of azeotropically dried sulfone
ER-804028 (1.0 wt., 1 eq.) in anhydrous tetrahydrofuran (5 vol.,
4.45 wt.) was treated with n-butyl lithium (1.6M in hexanes, 1.02
wt., 1.5 vol., 2.05 eq.) such that the internal temperature did not
exceed 5.degree. C. The mixture was stirred at internal temperature
0 to 5.degree. C. for 10 minutes then cooled to .[..rarw.75.degree.
C..]. .Iadd.<-75.degree. C. .Iaddend.Azeotropically dried
aldehyde ER-803896 (1.07 wt., 1.23 eq.) was dissolved in anhydrous
hexanes (3.53 wt., 5.35 vol.) then cooled to .[..rarw.75.degree.
C..]. .Iadd.<-75.degree. C. .Iaddend.The aldehyde solution was
added to the ER-804028 anion by cannula such that internal
temperature .ltoreq.-65.degree. C. The mixture was stirred for 45
minutes at internal temperature -78.degree. C. then quenched by the
addition of saturated ammonium chloride (5 vol.), methyl tert-butyl
ether (10 vol.), and water (5 vol.). The aqueous layer was
discarded and the organic layer concentrated under reduced
pressure. The crude material was purified via C-18 reverse phase
chromatography to afford ER-804029 (84%, 1.57 wt.).
##STR00146##
Sulfone-diol ER-804029 (1 wt., 1 eq.) was dissolved in wet
dichloromethane (7.4 vol., 0.04 wt % water) and placed in a
20-25.degree. C. water bath. Dess-Martin Reagent (0.67 wt., 2.5
eq.) was added in one portion. The reaction mixture was quenched
with saturated sodium bicarbonate (10 vol) and 10 wt % aqueous
sodium sulfite (10 vol.) and stirred for 30 minutes. The mixture
was diluted with saturated sodium chloride (10 vol) and extracted
with MTBE (25 vol). The aqueous layer was discarded and the organic
layer concentrated and purified by silica gel chromatography to
afford ER-804030 (0.9 wt., 90%). The material was stored under
inert gas atmosphere at -20.degree. C.
##STR00147##
To a pre-dried reactor under inert atmosphere was charged samarium
diiodide solution (2.5 eq.) and the solution cooled to internal
temperature .[..rarw.70.degree. C..]. .Iadd.<-70.degree. C.
.Iaddend.ER-804030 (1 wt.) was dissolved in anhydrous methanol (4.1
wt.) and anhydrous THF (2.3 wt.) and then cooled to
.[..rarw.70.degree. C..]. .Iadd.<-70.degree. C.
.Iaddend.ER-804030 was added to the cold samarium solution at a
rate such that the internal temperature did not exceed -70.degree.
C. The reaction was quenched with potassium carbonate/Rochelle's
Salts/water (1/10/100; w/w/v, 15 vol.) and MTBE (5 vol.) such that
internal temperature did not exceed -65.degree. C. Upon complete
addition of the workup solution, the reaction was warmed to room
temperature and the mixture transferred to a separatory vessel
using the workup solution (20 vol. rinse) and MTBE (20 vol. rinse).
The aqueous layer was discarded, the organic layer evaporated, and
the residue purified via silica gel chromatography to afford
ER-118049 (0.77 wt., 85%). The product was stored at -20.degree. C.
under inert atmosphere.
##STR00148##
A pre-dried reactor was charged with (S)-ligand ER-807363 (2.05 wt)
and the atmosphere was exchanged for nitrogen. The CrCl.sub.2 (0.85
wt, 10 eq) was added in one portion followed by anhydrous
acetonitrile (21.5 wt) and the mixture was warmed and maintained
between 30.degree. C. to 35.degree. C. Triethylamine (0.7 wt, 0.96
vol, 10 eq) was added in one portion and the mixture stirred for
one hour. The NiCl.sub.2 (0.09 wt, 1 eq) was added in one portion,
followed by the keto-aldehyde ER-118049 in anhydrous THF (2.43 wt,
2.73 vol) over 30 minutes. The heat was removed then heptane (20.5
wt, 30 vol) and Celite.RTM. (1.5 wt) were added. The mixture was
stirred for 5 minutes and filtered over a pad of Celite.RTM. (15
wt) and the Celite.RTM. pad rinsed with heptane (7.3 vol) and
acetonitrile (5 vol). The filtrate was transferred to a separatory
funnel and the lower layer removed. The combined heptane layers
were washed with acetonitrile (maximum 47.2 wt, maximum 60 vol) as
necessary. The heptane layer was evaporated under reduced pressure
and the product purified by silica gel chromatography to afford
ER-118047/048 (0.64 wt, 70%).
##STR00149##
Allyl alcohol ER-118047/048 was dissolved in dichloromethane (0.04
wt % water, 9 vol) and the reactor was placed in a water bath
(20.degree. C.) and the solution was treated with Dess-Martin
Reagent (0.48 wt, 1.5 eq). The reaction mixture was treated with
saturated aqueous sodium bicarbonate (9 vol) and 10 wt % aqueous
sodium sulfite (9 vol) then stirred for 20 minutes and transferred
to a separatory funnel with DCM (10 vol). The aqueous layer was
discarded, and the organic layer evaporated to a residue. The crude
material was purified by flash chromatography (prepped with 3 CV
(1:1 (V/V) DCM/heptane, the material was loaded with 1:1
DCM/heptane then eluted with 10/10/1 heptane/DCM/MTBE). The
product-containing fractions were concentrated and stored under
inert atmosphere at -20.degree. C.
##STR00150##
Alternatively, the oxidation of ER-118047/48 to form the di-ketone
ER-118046 was accomplished as follows. A flask was charged with
ER-118047/48 (1 wt, 1.0 eq) and toluene (10 vol) and DMSO (0.15
wts, 2.5 eq) were added at room temperature. Et.sub.3N (0.31 wts,
4.0 eq) was added and the solution was cooled to -15.degree. C.
TCAA (0.33 wts, 1.4 eq) was added neat and the reaction warmed to
0.degree. C. then stirred for 10 minutes at 0.degree. C. The
reaction was stirred for additional 10 minutes then was quenched
with IPA (0.15 vol). The reaction was stirred at 0.degree. C. for
10 minutes. 1N HCl (5 vol) was added over 2 minutes, and the
reaction was warmed to room temperature and diluted with MTBE (5
vol). Two clear layers formed and the aqueous layer was removed and
discarded. The organic layer was washed with 5 vol of 5%
bicarbonate (aqueous), concentrated to a heavy yellow oil on a
rotary evaporator and purified by silica gel chromatography (91%
isolated yield).
##STR00151##
Into an appropriately sized reaction vessel (vessel A) was charged
imidazole hydrochloride (0.39 wt, 5 eq) followed by 1 M TBAF in THF
(7.6 vol, 10 eq) at ambient temperature. The resulting mixture was
stirred until it is homogenous (15-30 minutes). Into a second
reaction vessel (vessel B) was charged ER-118046 (1 wt, 1 eq) and
THF (33 vol). The contents of vessel B were placed under an inert
atmosphere and stirred until ER-118046 was fully dissolved. The
contents of flask A (TBAF/Imidazole) were charged as a single
portion into flask B (ER-118046/THF). After 3-4 days, the reaction
solution was loaded onto a column and purified by silica gel
chromatography.
##STR00152##
The dried ER-118064 (F-12 wherein R.sup.1 is MeO) residue was
dissolved in anhydrous dichloromethane (28 vol) under a nitrogen
atmosphere and treated with PPTS (1.0 wt, 5.2 eq) in one portion.
After 30-90 minutes, the reaction mixture was directly loaded atop
an appropriate column and purified by silica gel chromatography.
The desired fractions of ER-076349 were concentrated in vacuo. The
material resulting from the concentration of all pure fractions was
azeotroped twice from toluene (20 vol), affording ER-076349 as a
crunchy colorless solid/foam (0.44 wt, 0.79 eq after correction for
residual toluene).
##STR00153##
In a clean dry reaction vessel (flask C) ER-076349 (1 wt, 1 eq) was
dissolved in anhydrous toluene (20 vol) and concentrated to dryness
under reduced pressure. The substrate was re-dissolved in anhydrous
toluene (20 vol) and concentrated to dryness. The substrate was
dissolved in DCM (5 vol), and the solution placed under an argon
atmosphere. Collidine (0.66 wts, 4.0 eq) was added as a single
portion. Pyridine, as a solution in DCM (Flask B), was added as a
single portion (5 mole %). The resulting mixture in flask C was
cooled to an internal temperature of -20 to -25.degree. C. A DCM
solution of Ts.sub.2O was added drop-wise keeping the internal
temperature below -16.degree. C. (1.02 eq). The reaction was
stirred at -20 to -25.degree. C. for 80 minutes then warmed to
0.degree. C. over 20 minutes and stirred for an additional 20
minutes. The reaction was quenched with water (2 vol). The bath was
removed, and the reaction allowed to warm to room temperature
(15-20.degree. C.) and stirred (20 minutes). The reaction was
rinsed to a larger vessel using the IPA (100 vol) and aqueous
ammonium hydroxide (100 vol) was added to the reaction. The
reaction was stirred at room temperature for 15-36 hours,
monitoring for the disappearance of the tosylate (ER-082892) and
epoxide (ER-809681) which formed in situ. The reaction was
concentrated to dryness or near dryness at reduced pressure. The
resulting material was diluted with DCM (25-40 vol) and washed pH
10 buffer (NaHCO.sub.3/Na.sub.2CO.sub.3 (aq), 10 vol). The aqueous
phase was back extracted with 25 vol of DCM and the combined
organic layers were concentrated to dryness. The resulting free
amine was purified by silica gel chromatography using a buffered
ACN/water mobile phase. The pooled fractions were concentrated at
reduced pressure to remove ACN. The resulting aqueous layer was
diluted with DCM (40 vol) and with 30 vol of a pH 10 buffered stock
solution (NaHCO.sub.3/Na.sub.2CO.sub.3). The layers were mixed well
and separated. The aqueous phase was back extracted with 25 vol of
DCM and the combined organic layers were concentrated to dryness.
The resulting free amine was polish filtered as a solution in 3:1
DCM/pentane and concentrated to dryness (0.80 wts) to afford
B-1939.
While we have described a number of embodiments of this invention,
it is apparent that our basic examples may be altered to provide
other embodiments that utilize the compounds and methods of this
invention. Therefore, it will be appreciated that the scope of this
invention is to be defined by the appended claims rather than by
the specific embodiments that have been represented by way of
example.
* * * * *