CA2418458A1 - Total synthesis of 14 beta-fluorosteroids via the transannular diels-alder reaction - Google Patents

Abstract

This work describes the total synthesis of 14.beta.-fluorosteroids via the Diels-Alder transannular reaction (DATA). All the synthetic routes tested are presented as well as all the intermediates obtained. First (chapter 1), we synthesized a thirteen-member macrocycle comprising olefins of trans-cis-cis geometry (TCC). After the key DATA reaction, a steroid tetracycle is obtained with the geometry at the junctions of the desired trans-synisic formed cycles (TSC). Chapter 2 describes all the attempts to homologate the ketone at position C-I7 and the formation of methyl at position C-13 from the steroidal tetracycle TS C. Chapter 3 describes the functionalization of cycle A to obtain the desired steroid skeleton and Chapter 4 describes the combination of these two model studies to obtain the 14.beta. - targeted fluorosteroids. Finally, chapter five describes the synthesis of 14.beta.-fluorosteroids from digitoxin 2 (natural compound) as well as the biological activities obtained for the 14.beta.fluorosteroids synthesized.

Description

Thanks First, I would like to thank Professor Pierre Deslongchamps for to have given me the opportunity to carry out my graduate studies in his laboratory what got me allowed to discover organic synthesis. I also thank him for his precious advice and for the freedom he left me throughout my studies which has greatly contributed to my scientific development.
I also thank all my laboratory colleagues with whom I have had pleasure in working. I
would like to thank in particular the members of the 2008: Olivier Lepage, Franck Caussanel, Serge Phoenix and François Bilodeau with whom the working atmosphere was always to his favorite. Special thanks to Julie Belzile, Julie Germain and Dominique Lambert who have become more than co-workers and have always been present for me.
I also thank Brigitte Poirier and Ian Levac who were two students that I had the chance to supervise and with whom I had fun working.
I would like to thank the support staff: Gaston Boulay (spectroscopy of mass), Marc Drouin and Andreas Decken (X-ray diffraction) and Normand Pothier (RMN).
Thank you also to Marie-Marthe Leroux for all the services rendered.
Thanks to the foundation of the University of Sherbrooke for the financial assistance brought and to the Neokimia company for biological tests.
Finally, a special thank you to all my family members and special to my mother Héléne and my father Denis who have always been present since the beginning of my studies and who have always encouraged me. Special thanks also to Jean Marc who was present all along of my studies and who now shares my life. Thank you for having me listened, helped, encouraged and advised.

Contents Summary................................................. ....., ................
.................................................. .......... i Thanks................................................. .................
.................................................. ..... ü
Table of materials ................................................. ......................
..........................................., iii List of schemes ................................................. .......................
............................................. vi List of paintings................................................. ......................
............................................. ix List of FIGS ................................................. .......................
................................................ x List of Abbreviations ................................................. ..................
.......................................... xü
Introduction................................................. ..................
.................................................. ....... 1 S
1. General information digitoxigenin ................................................. ................
................ 15

2. Biological activities of 14,13-hydroxysteroids and its analogues ......................... 16

3. The bioisosteric fluor of the hydroxyl group .................................................. ......... 17

4. Compounds Target ................................................. ........................
................................. 18

5. General information on the Diels-Alder reaction .................................................. ................ 19 5.1 Diels-Alder reaction intermolecular ................................................. ............

5.2 Intramolecular type Diels-Alder reaction 1 ............................................. 21 5.3 Type 2 intramolecular Diels-Alder reaction ............................................. 22

6. Description of project................................................. ........................
.......................... 23 6.1 Characterization of the molecule .................................................. ............................

6.2 Result earlier ................................................. ....................
............................ 23 6.3 Synthetic approach .................................................. .............................
............ 24 Results and discussion .................................................. .............................
............................ 26 Chapter 1 ................................................. .............................
................................................. 26 Macrocycle synthesis and tetracycle formation .................................................. .... 26 1.1 Analysis retrosynthetic ................................................. ..............
............................ 26 1.2 Synthesis of the /. Keto ester 18: precursor for the reaction of macrocyclization ....... 28 1.2.1 Synthesis of / 3-keto ester 18 by the route short ................................................. . 28 1.2.2 Synthesis of the ~ keto ester by the long route AT...............................................,. . 31 1.2.3 Synthesis of the ~ keto ester via the long route B
.................................................. 37 1.3 Macrocyclization and Transannular Diels-Alder Reaction ............................... 41 1.4 Chiral transannular Diels-Alder reaction .................................................. ...... 43 1.4.1 Previous results .................................................. ......., .................................. 43 1.4.2 Synthesis of the macrocycle and formation of the tetracycle ........................................... 44 1.4.3 Reducers Chiral ................................................. ................, ........................... 49 ...........
Chapter 2 ..
.................................................. .............................
........................... 53 . .......
Approval at position C-I7 .................................................. .............................
...... 53 2.I Approval of carbonyl at position C-17 ........ ......................................... ...... 53 2.2. Approval of carbonyl at position C-17 with reactions of coupling ...... 60 2.3 Formation of methyl at position C-13 and approval of the ketone at the position VS-................................................. 17 ............................
.................................................. .. 64 2.4 Compounds phenolic ................................................. ...................
.......................... 68 Chapter 3 ................................................. .............................
................................................. 71 Functionality of cycle A
.................................................. .............................
........... 71 3.1 Synthesis of the model for cyclopropanation .................................................. ....... 72 3.2 Cyclopropanation reaction with the mixture of alcohols 106 ................................. 74 3.2.1 Synthesis of alcohol 13 ................................................ ............................
............ 74 3.2.2 Reaction of cyclopropanation ................................................. ..............
............. 76 3.3 Alternative method of introducing methyl at position C-10 .................................. 79 3.4 Functionalization of cycle A with the proton at position C-10 ............................. 81 Chapter 4 ................................................. .............................
................................................. 85 Combination of the results obtained for the functionalization of the cycle A .................. 85 and for I homologation at position C-................................................. 17 ...................... 85 4.1 Approval of the ketone at position C-17 with cycle A
functionalized ......... 85 V
4.2 Functionalization of cycle A with approval made at position C-17 ............. 90 4.2.1 Strategy with nitrite at position C-................................................. 17 ........ 90 4.2.2 Strategy with the ester at position C-17 ............ .......... .......................... .......... 93 4.3 Strategy with 1 vinyl iodide and the functionalized cycle A
................................. 98 4.4 Synthesis of butenolide at position C-17 ................................. ............... ............. 102 4.4.1 Synthesis of butenolide by Stork ................................................. .................... 102 4.4.2 Synthesis of butenolide from nitrite .................................................. ...... 103 4.4.3 Synthesis of butenolide from 1 aldehyde ................................................. 104 4.5 Synthesis of analogues at position C-17 .................................................. ........... 108 Chapter ................................................. 5 .............................
............................................... 115 Synthesis of 14 /. ~ Fluorodigitoxigenin and activities biological ................................ 115 S.1 l4 ~ Fluorodigitoxigenin .................................................. .............................
........ 115 5.2 14 /. ~ Fluorodigitoxin .................................................. .............................
............... 119 5.3 Analog at position C-17 with fluorine at position C-14 .............................., .. I21 5.4 Biological activities .................................................. .............................
............... 122 General conclusion .................................................. .............................
............................. 127 Experimental part .................................................. .............................
............................. 128 General ................................................. ...................
.................................................. ...... 128 modes operating ................................................. ...................
............................................ 130 NMR spectra H ................................................. .............................
................................... 209 Bibliography................................................. .................
.................................................. .... 290 List of diagrams Diagram 1 ................................................. ....., .......................
.................................................. .24 Diagram 2 ................................................. .............. ..............
........ ......................................... . 25 Diagram 3 .................................................. .............................
............................................. .... 27 Diagram 4 ................................................. .............................
................................. ................ . 30 Diagram ................................................. 5 .............................
.................................................. . 32 Diagram 6 ................................. ............... .............................
......................., .......................... . 33 Diagram

................. 7, ............................... .............................
.................................................. . 35 Diagram

......................................... 8 ....... ........................ ....
.................................................. . 36 Diagram

................................................. 9 .............................
.................................................. . 37 Diagram

................................................. 10 ........................ ...
.................................................. 38 Diagram

................................................. 11 ... ........................
.................................................. 40 Diagram

................................................. 12 ............................
.................................................. 42 Figure 13 .................................................. .............................
................................................ 42 Diagram ................................................. 14 ............................
.................................................. 45 Diagram ................................................. 15 ............ ...............
......................., .......................... 47 Diagram ................................................. 16 ............................
.................................................. 49 Diagram ................................................. 17 ............................
., ................................................ 50 Diagram 18 .................................... ............ ............................
.................................................. 54 Diagram ................................................. 19 ..........................,.
.................................................. 55 Diagram ................................................. 20 ., ..........................
.................................................. 56 Diagram ................................................. 21 ............................
.................................................. 58 Figure 22 .................................................. .., ..........................
....................., .......................... 59 Figure 23 .................................................. .............................
................................................ 61 Diagram ................................................. 24 ............................
.................................................. 62 Figure 25 .................................................. .............................
................................................ 63 Figure 26 ............................. .................... ........................ ....
................................................ 65 Diagram ................................................. 27 ............................
..............................: ................... 66 Figure 28 .................................................. .............................
................................................ 67 Diagram ................................................. 29 ............................
.................................................. 69 Diagram 30................................................. ............................
.................................................. 71 Diagram 31 .......................................... ...... ............................
......................: ........................... 73 Diagram ................................................. 32 ............................
.................................................. 74 Figure 33 ......................................, ........... .............................
............, ................................... 76 Diagram ................................................. 34 ............................
.................................................. 77 Diagram ................................................. 35 ............................
.................................................. 80 Diagram ................................................. 36 .............., .............
.................................................. 82 Diagram ................................................. 37 ............................
.................................................. 86 Diagram ................................................. 38 ............................
.................................................. 89 Diagram ................................................. 39 ............................
.................................................. 91 Diagram ................................................. 40 ............................
.................................................. 91 Diagram ................................................. 41 ............................
.................................................. 93 Diagram ............................. 42, ................... ............................
.................................................. 95 Figure 43 .................................................. .............................
................................................ 96 Diagram ......................................... 44 ....... ............................
..........., ...................................... 99 Diagram 45 ............... ................................. ............................
................................................ 100 Diagram ................................................. 46 ............................
................................................ 101 Diagram ................................................. 47 ............................
................................................ 102 Figure 48 .................................................. .............................
.............................................. 104 Diagram ................................................. 49 ............................
................................................ 105 Diagram ................................................. 50 ............................
................................................ 106 Diagram 51 ........................., ....................... ............................
................................................ 107 Diagram ................................................. 52 ............................
................................................ 108 Figure 53 .................................................. .............................
.............................................. 109 Diagram ................................................. 54 ............................
................................., .............. 110 Figure 55 .................................................. .............................
.............................................. 111 Diagram ................................................. 56 ............................
................................................ 112 Diagram ................................................. 57 ............................
................................................ 113 Diagram 58 .............................., .................. ..........., ................
., .............................................. 114 Diagram .......................... 59, ...................... ............................
................................................ 116 Diagram 60 ............................................. ... ......................... ..
................................................ 117 Diagram ................................................. 61 ............................
................................................ 120 Diagram ................................................. 62 ............................
................................................ 121 Figure 63 .................................................. .............................
.............................................. 122 Diagram ................................................. 64 ............................
................................................ 124 List of paintings Table 1: Reaction conditions and yields for protection alcohol 45 ............. 38 Table 3: Alcohol protection conditions and yields of macrocycle 59. ............. 46 Table 4: Ratios obtained for the reaction of DATA on the macrocycles 60a 60f. ........ 48 Table 5: Nantioslective reduction of the macrocycle ................................................. 15 ...

Table 6: Reaction conditions for the approval of the.
ctone 75 and alcohol 74. 57 Table 7: Reaction conditions for approval of the tone 72. ............................ 59 Table 8: Reaction conditions for approval of the ctone 14. ......, ..................... 66 Table 9: Cyclopropanation tests on the mixture of alcohols 106. ................................. 78 Table 10: Reaction conditions for synthesizing ether d'nol triflique 120. ............... 87 Table 11: Reaction conditions for the approval of tone 116. ........................ 88 Table 12: Reducers used to pass nitrite 90b.
the corresponding aldhyde ..... 92 Table 13: Carbonylation tests on triflic nol ether.
88. ................................... 94 Table 14: Conditions for reducing the mixture of nitriles 106 147 .......................................

Table 15: Conditions for the protection of the fluorine compound 160. 120 ......................................

Table 16: Conditions for the preparation of reagents and 128 anhydrous solvents ...................

X
List of Figures Figure 1: Structures of digitoxigenin 1 and digitoxin 2.
......................................... 15 Figure 2: Structure of digoxin ....................................... 3, ......... ..........., .................
........ 16 Figure 3: Structures of digitoxigenin 1 and its analogs at position C-17 and their activities biological ................................................. .... ..............
........................... 17 Figure 4: Structures of targeted compounds.
.................................................. ..........................

Figure 5: Intermolecular Diels-Alder reaction.
.................................................. .......... 19 Figure 6: Overlay of border molecular orbitals for the reaction from Diels-Intermolecular alder.
.................................................. .............................
.................... 20 Figure 7: Endo and exo approach of the dienophile during a Diels- reaction Alder Transannular.
.................................................. .............................
.................................. 21 Figure 8: Examples of intramolecular Diels-Alder reactions.
...................................... 22 Figure 9: Diels-Alder reaction Transannular ............................................ .... ................

Figure 10: Structure of l4, Qfluorodigitoxigenin 7.
.................................................. ...... 23 Figure 11: Transannular Diels-Alder reaction on macrocycle 15.
........................... 26 Figure 12: The%. ~ Keto ester 18: precursor for the reaction of macrocyclization. ................ 28 Figure 13: Route used for the synthesis of 14 ~ -hydroxysteroids by DATAiI ................ 29 Figure 14: Mechanism for the Wittig reaction with fluorophosphonate 28.
............. 34 Figure 15: Transition state during the Diels-Alder reaction Transannular. ................... 43 Figure 16: Pascal Langlois 'previous results for Diels' reaction Alder Transannular. ........... ..............................., ..
....................... ............. .......... .... .... .............. 44 Figure 17: Transition states for the Diels-Alder reaction Transannular. ..................... 46 Figure 18: Aldehyde approval at position C-17 and formation of methyl at position C-

................................................. 13 ............................
...................................... 53 Figure 19: Attack face on nitrilate 82.
.................................................. ...................... 62 Figure 20: X-ray structure of nitrile 90b.
.................................................. ......, ............... 68 Figure 22: Structure of 1a10-nor-14% ~ fluorodigitoxigenin 117.
......................................... 84 Figure 23: Plan A for the synthesis of 10-nor-14 /. ~ Fluorodigitoxigenin 117 and its analogous to position C-17.
....,., ........................................... .............................
........... 85 Figure 24: Plan B for the synthesis of 10-nor-14, (~ fluorodigitoxigenin 117 and its analogous to position C-17.
.................................................. .............................
........... 90 Figure 25: Protonation of nitrilate 133 and enolate 135.
................................................. 97 Figure 26: X-ray structure of nitrite ~ 143.
.................................................. ................. 101 Figure 27: Reduction of the nitrile mixture 147.
.................................................. ............ 105 Figure 28: X-ray structure of 14% j-fluorodigitoxigenin 7.
........................................ 118 Figure 29: Front side of the fluoride................................................. ......................
.......... 118 Figure 30: Structure of 1 folic acid 165 and methotrexate 166 ................................... 125 Abreviations list At Angstrom Absolute abs Ac Actyle add Addition APTS p-tolunesulfonic acid BSA N O-Bistrimthylsilylactamide BHT 2,6-di-t-butyl-4-phnol Bn Benzyle Bu Butyle C Degr Celcius conc Concentr CST cis-syn-trans DAST (dithylamino) sulfide trifloride DATA Diels-Alder transannulaire dcomp Dcomposition DEAD Dithylazadicarbonylate Dibal dsobutylaluminium hydride DIPEA Dsopropylthylamine DMAP 4- (N, N Dimthylamino) pyridine DMF N, N Dimthylformamide DMS Dimthylsulfide DMSO Dimthylsulfoxide E Methyl ester EE thoxythyle q equivalent And thyle EVE thylvinylther g Gram xill g Gram h Time HCA Hexachloroactone hex Hexane HMPA Hexamthylphosphoramide HOMO The highest molecular orbital occupies Hz Hertz ICso Concentration to obtain 50% inhibition Im imidazole IR Infrared J Coupling constant KHMDS Bis (trimthylsilyl) potassium amide LDA Lithium dsopropylamide lut lutidine LUMO The lowest unoccupied molecular orbital m Mta M Molar m-CPBA m-chloroperoxybenzoque acid Me Mthyle mg Milligrams min Minute mL Milliliter mmol Millimole MOM Mthoxymthyle Ms Mthanesulfonyle N Normal NaHMDS Bis (trimthylsilyl) sodium amide N N oxide of methyl morpholine p Para pent pentane Piv Pivaloyle PPTS pyrridinium p-Tolunesulfonate pyr pyridine quat Quaternary Yield yield NMR Nuclear magnetic resonance t Third your ambient temperature TBAF Ttrapropylammonium fluoride TBDMS t-Butyldimthylsilyl TBDMSOTf t-Butyldimthylsilyl trifluoromthanesulfonate TBDPS t-Butyldiphnylsilyl TCC trans-cis-cis tertiary Tertiary Tf Trifluoromthanesulfonate THP Ttrahydropyrane THF Ttrahydrofuran TIPS Trsopropylsilyl TIPSOTf Trsopopylsilyl trifluoromthanesulfonate TMS Trimthylsilyl TMSOTf Trmthylsilyl trifluoromthanesulfonate TPAP Ttrapropylammonium Perruthnate Tr Trityle Ts Tolunesulfonyl TSC Trans-syn-cis micromolar 12-C-4 ther crown I2-C-4 18-C-6 ther crown 18-C-6 Introduction 1. General information on digitoxigenin Digitoxigenin 1 is a natural heart glycoside extracted from the plant digitalis purpureal.
This plant contains a mixture of several types of cardiac glycosides which vary according to the locality of the latter as well as the season. Digitoxigenin 1 is part of the big family of digitalis also called carbenolides and it is the most compound prevalent of this families. Digitoxigenin 1 comes from the reaction hydrolysis of sugar to position C-3 of digitoxin 2 (Figure 1).
O
1 h Figure 1: Structures of digitoxigenin 1 and digitoxin 2.
Compounds of the digitalis family differ from ordinary steroids in three main pointsl. First, the junction of the CD cycles is cis instead of being trans. Second, there is an alcohol at the C-14 position of these steroids and finally the substituting for position C-17 (a butenolide) is in its configuration the less stable either / 3 instead of a.

2. Biological activities of 14 / .x-hydroxysteroids and its analogues The compounds of the digitalis family were used for the first times in 1785 by William Withering for the treatment of heart diseasel. The l4, Qhydroxysteroids are therefore used for many years as a pacemaker, as expectorant and as a diuretic. They are still prescribed nowadays for treatment diseases Cardiovascular. Digoxin 3 is the most prescribed compound and is differentiates from the digitoxin 2 by its hydroxyl group at position C-12 of the skeleton (figure 2).

O
HO
OH
Figure 2: Structure of digoxin 3.
The main biological activity of cardiac glycosides is their capacity to increase the force of myocardial contractions (positive inotropic effectj. This effect inotropic positive results from the inhibition of the membrane enzyme Na +, K + -ATPase responsible for maintenance of the high concentration of Na + and K + in the intracellular fluid. grace to a study of the structure-activity relationship, it has been determined that the hydroxyl group at position C-14 of 14, f ~ hydroxysteroids is essential for the activity of these compounds2. Through against this study has not demonstrated whether the hydroxyl group at this position is a acceptor or a bridge donor H.
It turns out that the compounds of this family are very toxic because the dose therapeutic can match up to 60% of the lethal dose. The therapeutic dose for processing 1 ~%
cardiovascular disease is extremely small at 0.2 mg for the 2 'digitoxin. So, since 1960 many efforts have been made to synthesize analogs of these carbenolides in an attempt to decrease the toxicity of these products while keeping good biological activity. Figure 3 shows digitoxigenin 1 and analogs of the latter at position C-17 (compounds 4 to 6) as well as their biological activities3. We can see that by changing the chain at position C-17, biological activities remain comparable to that of digitoxigenin 1 for compounds 4 and 5 with the chain / ~ (ICSO
equal to 0.35 and 0.03 ~ M respectively instead of 0.08 ~ M) but when the chain is a, biological activity decreases (compound 6, ICSO = 5.89 ~ M).
R

HH
4 C-17 ~, R = Me IC5o = 0.35 ~ M
1 ICSO = 0.08 ~ M 5 C-17 ~, R = CH2CH2NMe2 ICSO = 0.03 ~ M
6 C-17a, R = CH2CH2NMe2 ICSO = 5.89 yM
Figure 3: Structures of digitoxigenin 1 and its analogues to position C-1? as well as their biological activities.
3. The bioisosteric fluor of the hydroxyl group A bioisostere is a chemical group that can replace another in a drug without modifying the biological activity 4. It does not depend on the valence of groupings ni of their steric hindrance. When a group is replaced by a bioisostere of many parameters can change such as size, shape, electronic distribution, chemical reactivity and hydrogen bonds. When we change these parameters we can modify the structure of the molecule, its interactions with the receptor, its transport and sound metabolism.
Bioisosterism is an approach that is used to mitigate toxicity of a drug 4. he is important to note that there is no generalization because a good bioisostere in a branch of medicinal chemistry may become useless in another branch.
There are tables based on previous results4, indicating the relationships of possible bioisosterism between different groupings. However, it is activity retention organic which determines if two groups are bioisosteric and nothing else.
By consulting these tables, we have found that fluorine is a bioisostere of the group hydroxyle4. So by replacing the hydroxyl group at position C-14 with a fluorine, we can get the fluorinated analog of digitoxigenin 1. By this modification we could verify that the hydroxyl group, at this position of the digitoxigenin 1, is well an H bridge acceptor and not an H bridge donor because fluorine can only be a bridge acceptor H. Therefore, it seems very logical to introduce a fluor at position C-14. In addition, if we obtain an interesting biological activity for this fluorostéroïde, we can potentially get a new drug that might be less toxic.
4. Target compounds We have chosen as target compounds three new molecules which contain fluorine at position C-14 instead of the hydroxyl group (Figure 4). Compound 7 is the analog fluorinated digitoxigenin 1 and compounds 8 and 9 are also analogues of the digitoxigenin 1 but, in addition to having fluorine at position C-14, they have a ester a, ~ 3 unsaturated at position C-17 instead of butenolide. To synthesize these compounds fluorinated we want use the Transannular Diels-Alder reaction as a key step.

CO ~ Me 7 8 C-17 ~
9 C-17a Figure 4: Structure of the targeted compounds.
5. General information on the Diels-Alder reaction Diels-Alder's reaction was discovered in 1928 by Otto Diels and Kurt Alders and it's a cycloadditions which is the most important in organic chemistry. there is three types of Diels-Alder reaction: the intermolecular version and two versions intramolecular, a type 1 and the other type 2 (Diels-Alder transannulaire).
5.1 Intermolecular Diels-Alder reaction i: .a Diels-Alder intermolecular reaction is to react a Die ~ = '<stepped under her cisoid form and an alkene called dienophile to give a cyclohexene in passing by a boat 6 transition state (figure 5). This reaction is also called a reaction of cycloaddition [4 + 2]. It involves the four electrons r ~ of the diene and the two electrons ~ of the dienophile. During the reaction, two ~ links are formed so concerted as well as a link + ~ '. -,, OJ
Figure 5: Intermolecular Diels-Alder reaction.

For the Diels-Alder reaction to be allowed, the overlap must be orbitals molecular boundaries be positive (binder). There are two types of reaction Diels-Alder. The first, which we call conventional Diels-Alder, involves the HOMO of diene and the LUMO of the dienophile and the second, which we call Diels-Alder with reverse request, fact intervene the LUMO of the diene and the HOMO of the dienophile (FIG. 6).
HOMO of the diene LUMO of the diene LUMO of the dienophile ~ --g HOMO of the dienophile Conventional Diels-Alder Reverse-demand Diels-Alder Figure 6: Overlay of border molecular orbitals for the reaction from Diels-Intermolecular alder.
The higher the HOMO of the diene, the higher the LUMO of the dienophile.
low in energy, the more the Diels-Alder reaction is favored. The presence of substituents electron-withdrawing or electron-donating diene and dienophile influence the energy of molecular orbital boundaries of these two entities. Indeed, a group electron increases the energy of HOMO and LUWO and an electron-withdrawing group decreases the energy of HOMO and LUMO. The best possible combination, for that the Diels-Alder reaction to happen, so is to have a substituent electro donor on the diene and an electron-withdrawing substituent on the dienophile to have the most little difference energy.
The intermolecular Diels-Alder reaction is stereospecific because the addition is syn in respecting the geometries of the diene and the dienophile. When the dienophile is substituted, we can have two possible attack faces on the diene. The dienophile can approach by endo side or by the exo side of the diene (Figure 7). By the endo side, the substituents of dienophile are oriented below the diene and by the exo side, they are exterior oriented of it. Endo preference is observed when the dienophile is conjugated with a electron-withdrawing group because there are orbital interactions favorable secondary which are developed '. .
RR
', ,, YY
H ~ ~ Endo approach RX
X
HR
RR
~, H ,, ~ Y
Y '~ Exo approach R ,, /., ~ X
H
XR
Figure 7: Endo and exo approach of the dienophile during a Diels- reaction Alder Transannular.
5.2 Type 1 intramolecular Diels-Alder reaction In the case of a type 1 intramolecular Diels-Alder, the diene and the dienophile belong to the same acyclic molecule. The same orbital rules molecular boundaries, as those for the Intermolecular Diels-Alder, apply. Through against, in the in most cases the intramolecular version of Diels-Alder is much more stereoselective and regioselective. This is explained by the fact that the diene and the dienophile attached together and that the cycle can favor one conformation over another.
This preferably depends on the length of the chain between the diene and the dienophile.
Cyclization can therefore lead to a merged system (favored with small chains) or good to one bridged system (with chains of more than ten carbons) as presented in the figure 8.

Merged system Bridged system Figure 8: Examples of intramolecular Diels-Alder reactions.
5.3 Type 2 intramolecular Diels-Alder reaction The intramolecular type 2 Diels-Alder reaction involves a diene and a dienophile which are found inside a cycle called macrocycle. This reaction is also called a transannular Diels-Alder reaction (DATA). This reaction therefore has a higher degree of restriction than the intramolecular version of type 1. By therefore she is more stereoselective and regioselective. The DATA reaction is characterized through training of polycyclic products and by the simultaneous creation of four new centers asymmetrical (Figure 9).
DATA
Figure 9: Transannular Diels-Alder reaction.
The tansansular Diels-Alder reaction is therefore a very powerful reaction to train polycyclic products such as steroids. Our laboratory is interested in this type of reaction for several years and many studies have been do on this dernière8-9. According to the stereochemistry of olefins inside the macrocycle, we can obtain several polycyclic compounds with different stereochemistry and perfectly checked at the junctions of the cycles formed.
6. Description of the project 6.1 Characterization of the molecule We therefore set ourselves the objective of synthesizing the 14 ~ -fluorodigitoxigenin 7 (figure 10). It is a steroid with stereochemistry at the junctions of B- cycles CD is trans-syn-cis (TSC ~. The junction between the AB cycle is cis and the molecule has an alcohol ~ 3 to the position C-3 and a fluorine / 3 at position C-14 instead of the hydroxyl which is in the compound natural. The chain at position C-17 is a butenolide of ~ 3 stereochemistry and there is a methyl at position C-10 and another at position C-13. We must therefore take into account of all these elements during the synthesis of 14, Q fluorodigitoxigenin 7.

Figure 10: Structure of 14, a fluorodigitoxigenin 7.
6.2 Previous results As mentioned earlier, the 14, ~ fluorodigitoxigenin 7 that we want synthesizing involves stereochemistry at the junctions of trans-syn- BCD cycles cis (TSC ~.
The results previously obtained in our laboratory (diagram 1) tell us demonstrated that by synthesizing a thirteen-member macrocycle, comprising the diene with the olefins of trans-cis geometry and the cis geometry dienophile we get a ABC tricycle [6.6.5]
and the stereochemistry at the junctions of the cycles formed is then trans-syn-cis (TSC). The first one model study was carried out by Pierre Soucyl ° and it shows that starting from the macrocycle to thirteen members 10, with the geometry of the TCC olefins, the tricycle 11 is then formed with the geometry at the junctions of the TSC cycles. In addition, the work of Luke Ouelletll have us demonstrated that from the TCC 12 macrocycle, the TSC 13 tetracycle is formed and it corresponds to the backbone of digitoxigenin 1 and its fluorinated analogue 7, with the stereochemistry adequate BCD cycle junctions.
Even Soucy (~ o C02t-Bu DATA ~ C02t-Bu / C02t-Bu C02t-Bu EJEH GOLD
EE

EOEO
Ouellet I \ o DATA
GOLD
me0 me0 Diagram 1 6.3 Synthetic approach The general strategy for synthesizing 143 fluorodigitoxigenin 7 is shown in the diagram 2. It involves the Transannular Diels-Alder reaction (DATA) to training BCD cycles. To carry out this reaction, it will be necessary to synthesize the thirteen macrocycle members 15 comprising a diene and a dienophile. The geometry of the doubles links of diene should be trans-cis (TC) and the geometry of the double bond of the dienophile will have to be cis (C). With this macrocycle 15 of TCC geometry, we will be able to obtain the tetracycle 14 with stereochemistry at the junctions of trans-syn-cis BCD cycles (TSG ~. For what is cycle A, it will already be present in its aromatic form from the beginning of the synthesis. There we it will therefore have to be reduced, introduce a methyl at position C-10 and obtain the cycle junction AB ci.ç. Finally, it will remain to synthesize the butenolide at position C-17 from ketone.
O
Me OEO
_ ~ F
I /
HF
Me0 ~ Me0 Diagram 2 Results and discussion Chapter 1 Macrocycle synthesis and tetracycle formation Chapter 1 describes the synthesis of tetracycle 16 (Figure 11), which has junctions of trans-syn-cis cycles (TSC ~, via the transannular Diels-Alder reaction (DATA) from macrocycle 15 whose geometry of the diene is trans-cis (TC ~ and that of dienophile is cis (G ~.
Three synthetic routes, one of which has not worked, will be described to get there to the precursor of the macrocyclization reaction. In addition, a summary enantioselective of tètracycle 16 will be proposed as well as the attempts to get there.

data_ HF
Me0 ~ Me0 Figure I1: Transannular Diels-Alder reaction on the m ~ zcrocycle 15.
1.1 Retosynthetic analysis The retrosynthetic analysis is presented in diagram 3. First, analog fluorinated at position 14 ~ i of digitoxigenin, that is to say compound 7, could be obtained at from tetracycle 17 by homologation of the ketone at position C-17 to form the butenolide group. Tetracycle I7 could be synthesized from compound 14 in going through a Birch reduction of cycle A followed by a series of reactions to get a cyclopropane at the junction of the AB cycles. This cyclopropane could then be open for obtain the methyl at position C-10.
OO
Me Me O Me O
Me ~ Me ti ~ F ~ ~, HF ~ (\, ~ F v O r RO Me0 i7 EO
'OPiv \ 'F
/ I ~ ~ I /
F
Me0 \ OMe Me0 J,%
'i, ~., .. OR O
- 'FC ~.%' / ~~~ OH
MeO ~ \ "~ i ~., ._ ~~ CO And Me0 \ I ~ OH MeO

1g 20 21 Diagram 3 For the formation of tetracycle 14 which includes trans- cycle junctions syn-cis (TSC), synthesis of the thirteen-member macrocycle 15 is necessary. The geometry of diene must be t ~ ° ans-cis (TC) and that of the cis dienophile (C). So from macrocycle 15 of TCC geometry we could get the tetracycle 14 with junctions of TSC cycles via a transannular Diels-Alder reaction (DATA). Macrocycle 15 could to be trained to from ~ keto ester 18 by a macrocyclization catalyzed by palladium.
This ~ keto ester could be obtained from compound 19 through a series of transformations of functional groups and by the alkylation of the ~ -keto ester portion. AT
from diol 20, we could get compound 19 by differentiating the two alcohols and by approving each of the two chains by Wittig reactions of the Horner-Emmonsl2 type with some different phosphonates. The fluorine found at position 143 in the final product would therefore introduced at this stage of the synthesis. Finally, Diol 20 could come from ketone 21 which is commercially available.
1.2 Synthesis of ~ keto ester 18: precursor for the reaction of macrocyclization I.2.1 Synthesis of ~ keto ester 18 by the short route At first, we tried to synthesize the ~ keto ester 18, which is the precursor for the macrocyclization reaction, by the shortest route in passing by a amide of Weinreb (figure 12). This channel is inspired by the one used previously in our laboratory for the synthesis of 14 ~ -hydroxysteroids carried out by Luc Ouelletl i (figure 13).
'OPiv macrocyclization - '~
Me0 ~ OMe ~ / J
me0 Figure 12: The, a keto ester 18: precursor for the reaction of macrocyclization.
In this synthesis, the dialdehyde 23 is prepared in three stages in performing a reduction ketone 21 followed by elimination of the corresponding alcohol and ozonolysis the olefin obtained. To differentiate the two aldehydes of compound 23, two reactions from Wittig of type Horner-Emmons'2 are performed. The first is selective to aldehyde aliphatic and the corresponding ester, ~ unsaturated is obtained in a yield 64% for the last four steps. Horner's second Wittig reaction-Emmonsl2 is then carried out on benzyl aldehyde to obtain compound 24 in a yield of 85%. In just five steps, the advanced intermediary 24 is obtained with excellent yields. There are only seven steps left to synthesize the ~ keto ester 25.
A series of changes in functional groups and an alkylation reaction of the portion, (i 'keto esters are then produced with very good yields (from 78% to 99%).
Finally in three other steps, including macrocyclization and Diels-Alder reactions transarmulaire, the tetracycle 26 is obtained.
OO ~ Rome % ~ ~ ~ N ~ Me 3 steps ~ \ IHH 2 steps ii OMOM
Me0 Me0 21 23 O Me0 24 C02And EO
~ OPiv 7 steps / ~ 3 steps ~ OMOM
, .J ~ \
MeO ~ ~~~ _- ~~ ~, / ~ - ~ .OMe ~ \ H OMOM
25 ~ ~ Me0 Figure 13: Route used for the synthesis of l4 ~ hydroxysteroids by DATA ".
To begin our synthesis, we repeated the first three 'steps of the sequence made by Luc Ouelletl '~' 4 (figure 13). We reduced ketone 21 available commercially with lithium aluminum hydride at -78 ° C for get alcohol corresponding. The latter is subject to the conditions of dehydration with p- acid toluenesulfonic in benzene at reflux to give the olefin 27 in a yield quantitative. This olefin is treated with ozone and then with dimethyl sulfide and we get Ie dialdehyde 23. The latter is very unstable even at low temperatures and under atmosphere inert gas. So we react immediately with the anion of the 28's fluorophosphonate and 26% of the ester a, l, unsaturated 29 is obtained in a mixture trans: cis 4: 1 (diagram 4). Several decomposition products are observed and the efforts to try to optimize the reaction were unsuccessful. In the case of Luc Ouellet described earlier the time of the Horner-Emmonsl2 type Wittig reaction on the same dialdehyde with a similar phosphonate (OMOM instead of F), was shorter and the reaction was performed at -78 ° Cl, 14. In our case, at -78 ° C, there is no reaction and by warming the environment reaction, the decomposition rate of dialdehyde is accelerated 23. We so let's get the ester has, ~ unsaturated 29 with a poor yield since the aldehyde is breaks down before to react with the anion of fluorophosphonate 28IS. So we didn't get the intermediary advanced 31 by this synthetic route.
O
/ 1} LiAIH4, THF, -78 ° C / ~ 1) 03, MeOH, -78 ~ C
2) APTS, benz, reflux ~ ~ 2) DMS, -78 ° C at ta Me0 100% Me0 n-BuLi, THF, -78 ° C to 0 ° C
-. - ~ .H
(Et0) P (O) CH (F) C02Et 28, F
H ~ ', ~,.
Me0 '' ~ 26%, 2 steps MeO ~ '~~ CO tt O 29 4: 1 t'rans: cis 23 unstable (Et0) 2PCH2oN'OMe N, OMe ~ Me ~ ~ Me ________ 3 ~ _____._ i /
_ I ~ F

Diagram 4 In an attempt to resolve the problem, we have tried to selectively protect aldehyde aliphatic of dialdehyde 23 by acetalization with ethylene glycol in using different reaction conditions. In most cases, a mixture of the product of monoprotection and diprotection product has been observed as well as decomposition. The best yield that we got for the monoprotection reaction of the aldehyde aliphatic of compound 23 is 35% using the method of Luchel6 (with ErCl3). This yield is not much better than that obtained during the reaction of Wittig who is 26% (figure 4). So we decided to drop this path of synthesis.
1.2.2 Synthesis of the! 3-keto ester 18 by the long route A
Given the low yields of the Horner-type Wittig reaction Emmonsl2 on the dialdehyde 23 with fluorophosphonate 2815 we used a synthesis more long to obtain the, ~ keto ester 18. This route is always inspired by that used by Luke Ouellet for the synthesis of 14,13-hydroxysteroidsli, ia (Figure 14). The amide from Weinreb 31 (diagram 4) is the first key intermediary targeted. Subsequently, the same steps accomplished by Luc Ouellet will be used on our fluorinated compounds to synthesize Ie % ~ keto ester 18 (figure 12).
First, we did the ozonolysis of the olétïne 27 obtained previously (diagram 4) and we treated the intermediate formed with borohydride sodiuml ~
(diagram 5). Then an oxidative treatment is carried out with hydroxide sodium 10% and 30% hydrogen peroxide to destroy boronate salts in solution18. We so gets diol 20 in a yield of 86%. We must now differentiate the two alcohols which are primary but one of which is benzyl9. So we treat diol 20 with dioxide manganese (IV) in a mixture of pentane and dichloromethane, conditions for change from an allylic or benzyl alcohol to the corresponding aldehyde. We then gets Hydroxyaldehyde 32 in 85% yield. It is important to note that this hydroxyaldehyde is unstable. Horner-Emmonsl2-type Wittig reaction with the phosphonate 302 °, which contains the amide from Weinreb, is therefore rapidly performed on this hydroxyaldehyde 32. We thus obtain the amine of Weinreb a, ~ unsaturated 33 in a 49% yield (Figure 5). Only the trans geometry of the olefin has been observed by NMR
H. This low yield is probably due to the presence of alcohol free hydroxyaldehyde 32 which can react with the anion of phosphonate 30 and so cause unwanted side reactions.
1) 03, MeOH, -78 ° C
2) NaBH4, ta ~ ~ I '' \ 0H Mn02 Me0 \ 3) NaOH, H202, THF Me0 ~ ° ~ OH Pent: CH2Cl2, 85%
27 g6% 20 O
, OMe .OMe O (Et0) 2PCH2CN I N'Me H 30 ~ Me OH NaH, Et20, 0 ° C, 49% ~ OH
Me0 Me0 Diagram 5 We therefore protected the alcohol from hydroxyaldehyde 32 with t- chloride.

butyldimëthyïsilane in the presence of imidazole to obtain aldehyde 34 and We have then repeat the Wittigtz reaction on this intermediate with the phosphonate 30z °. We thus we obtain the amide of Weinreb 35 in a yield of 85% for the two steps (diagram 6). The silyl ether of this compound is then treated with fluoride.
tetrabutylarnmonium to give alcohol 33 which was already obtained previously (diagram 5). This alcohol is oxidized under the conditions of SwernZ ~ to obtain the aldehyde 36 in one 94% yield. We are now ready to do Wittig's reaction Of type Horner-Emmonsl2 with flurophosphonate 285. We therefore react the anion of fluorophosphonate 28 with aldehyde 36 at a concentration of SxlO'ZM (otherwise the yield is lower) and the ester has "unsaturated Q 31 in a 88% efficiency (diagram 6). The isomer with olefin of trans geometry with respect to fluorine is the only one product observed by 1 H NMR.
OO
, - H TBDMSCI. imidazole, OH THF, 100% _H
Me0 Me0 ~ I '~ OTBDMS

OO
(Et0) 2PCH2CN'OMe IN ~ Mé e 30, Me _ / 1) TBAF, THF, 100%
NaH, Et20, 0 ° C, 85% ~ ~ OTBDMS 2) Swern, 94%
me0 OMe Me n_guLi, THF, -78 ° C to 0 ° C
(Et0) 2P (O) CH (F) C02Et 28 H
88%

Diagram 6 The geometric isomer of ~ 3 unsaturated keto ester 31, obtained during the reaction Wittig's Horner-Emmonsl2 type with fluorophosphonate 28, is explained in literature by Barton'S works. It has been shown that with the lithium ion, the E isomer predominates and that with sodium and potassium ions, there is almost no selectivity. This is explained with the mechanism of the Wittig reaction (Figure 14). The training of erythro intermediates and threo is reversible and both diastereoisomers may be present at the balance. When the lithium ion is complexed on these two intermediates, the first stage of the mechanism then becomes irreversible. There is therefore no more equilibrium between the isomer Erythro kinetics and the thermodynamic isomer threo. The ratio of olefins E and Z is by therefore controlled by the first step of the reaction. Since we are in conditions kinetics with the ion lithium, we obtain the isomer é ~ ythro in a majority way. The latter therefore breaks down in isomer E which is then mainly obtained. Not knowing nature exact transition states leading to the erythro and threo isomers it is difficult for the time being attempt to explain the preferential formation of the erythro isomer by kinetic wnditions.
In our case, we are only looking at the E isomer as the final product, the erythro intermediate is the only diastereoisomer formed.
O ~ FO ~~ F ~~ C02And (Et0) zP C02Et - ~ (Et0) zP C02Et / _ \ -~ O ~~ H Ö ~~ HHF
[(Et0) zP (O) CFC02Et] -l.i + ~ RR
erythro RCHO ~ O_ \ R
OFF "F
(Et0) zPO ~ R02Et - (Et0) zÖ ~ R02Et '~~ H ~ - ~ C02Et Z
threo Figure 14: Mechanism for the Wittig reaction with fluorophosphonate 28.
To continue the sequence, you have to reduce intermediary 31. The part amide of Weïnreb of the latter must be reduced to aldehyde and the ester part to adLO ~ if to can differentiate the two groupings. To do this, some reducers such as 1'hydc ~ ure aluminum and lithium, düsopropylaluminium hydride and borohydride lithium have been used. The best result has been obtained with aluminum hydride and lithium to low temperature. We thus obtain hydroxyaldehyde 37 in a weak yield of 44% (figure 7). Indeed, many secondary products are fbrmés during this reaction and they are very difficult to separate from the desired product. Despite this low yield we we continued the sequence until ~ keto ester 18. We will come back more late in the section of long track B, to describe how we got around the problem reduction of intermediary 31.

O
, OMe 'N ~ Me LiAIH4, THF IH
F -78 ° C to 0 ° C ~ i IF
C02And 44% MeO \ --OH

OH
HCA, PPh ~ VaBH4, MeOH
THF, 77% 77% 54% ~ F
Me0 Me0 THIS
$ 3 39 ~ OTBDMS
TBDMSCI, imidazol ~ r Nal, NaH, n-BuLi THF, 71% F MeC (O) CH2C02Me, Me0 CI THF, 76%
OTBDMS 1 Y TBAF T ~. 77 ° / a ~ ~~ OPiv 2) PivCl, 2,6-lut .F CH2CIl, 97 ~~ 0 ~ / ~~, =: ._: r ~ F
~ .neO ~ ~~. ~. ~ ..OMe Me0 '_. '~ ~~. ¿~. OMe 41 OO 1g OO
Diagram 7 The alcohol of compound 37 is converted into the corresponding chloride using the method from Magid22 which uses hexachloroacetone and triphenylphosphine. With this method, We have obtained chloride 38 in a yield of 77% (scheme 7). We reduce aldehyde 38 with the sodium borohydride and the resulting alcohol 39 is protected with sodium chloride t-butyldimethylsilane in the presence of imidazole to give compound 40 in a yield 38% for the two stages. We then move the chloride from this intermediary using the dianion of methyl acetoacetate in the presence of sodium iodide for get the keto ester 41 in 76% yield. This compound is treated with fluoride tetrabutylammonium and the corresponding alcohol is obtained in a yield of 77%. We transform this alcohol into a pivalo ester using 2,6-lutidine and chloride trimethylacetyl to lead to ~ 3 keto ester 18 in a yield of 97%
(diagram 7).
By this route, the ~ 3 keto ester I8 which is the precursor for the reaction of macrocyclization, a been synthesized in 1? stages with an overall yield of 3.8%. This yield will be compared later with that obtained for the synthesis of the same ~ keto ester by the way long B.
To try to improve the step of reducing intermediary 31 (diagram 7), We have reduces the Weinreb amide before the introduction of the ester group by the wittig reaction Homer-Emmonsl2 type with fluorophosphonate 3015. To do this, we have used intermediary 35 already obtained (diagram 6) and we made it react with the borohydride lithium. Alcohol 42 was obtained in a 70% yield with impurities since there a lot of unidentified side products that are very difficult to separate from this latest. We therefore reduced amide 35 with aluminum hydride and lithium for obtain the aldehyde 43 in a yield of 58% and 12% of the alcohol 42 (diagram 8).
OH ~
.OMe LiBH4, THF, ta ~ '~ .. ~~ DTBDMS
N.Me 70% (impure) Me0 ~~~ OTBDMS
LiAIH4, THF
ta, 58 ° /
35 + 12% alcohol 42 BDMS

Diagram 8 Not being satisfied with the results obtained for the reduction of amide 35, we needed find another way to get the ~ keto ester 18 (long route B).
1.2.3 Synthesis of the ~ -keto ester 18 by the long route B
To improve the synthesis of the ~ -keto ester 18 obtained in the long route A
(diagram 7), we have replaced the Weinreb amide of compound 37 (scheme 8) with an ester (compound 44) which would be easier to reduce (Figure 9). To do this, we started the sequence with the aldehyde 34 already obtained before (diagram 6) and we made it react with the anion of triethyl phosphonoacetate. Ester a ,, ~ unsaturated 44 of trans geometry exclusively, determined by 1H NMR, was obtained in a yield of 97%. Many sources hydrides such as aluminum and lithium hydride (65%), lithium borohydride (81%), sodium borohydride (84%) and düsopropylaluminium hydride (9?%) Were used to reduce the ester a, / ~ unsaturated 44. It is therefore the hydride of düsopropylaluminium which gave us the best performance. Now we have to find a protective group compatible with the silylated ether of compound 45 (TBDMS), that is to say stable in the deprotection conditions for the latter.
O
I ~
O t ~ ..OEt Nal ~, THF
\ H (Et0) 2P (O) CH2COzEt OTBDMS g7%, ~ I OTBDMS
me0 me0 ~ OH ~ ~ OR
Dibal, CH2CI2 ~ Prot_ection i 0 ° C, 97% ~ ~ w ~
OTBDMS ~ I OTBDMS
Me0 Me0 Diagram 9 The different protective groups that we examined as well as the conditions of reactions are listed in Table 1. First, the groupemenrs methoxymethyl (MOM) and tetrahydropyranyl (THP), stable in the presence of ions fluorides (usual conditions for deprotection of silylated ethers), were used to protect alcohol 4:
(inputs 1 and 2). The yields of these two reactions are relatively low (42% and 65%
respectively). So we decided to use t-butyldiphenylsilyl as protective group and the reaction yield is. significantly higher or 93%
(entry 3).
Board 1:
terms and yields reaction for protection alcohol 45.

~~~ Enter ~~ W ~~~~~~~ Reaction conditions Yield v Compos R ~ Y- ~ -___......._..__...._..._..___._...._._......._.__......_ .....___..._....___._._ __....__._ ~, ~ .._____.____.._.______._._._______ .___.____.___ ~ .__ _____ ~ .... ~ ______.__-....___..______..._._.._ 1 MOM MOMCI, DIPEA, CHZClz 42% 46a 2 THP DHP, APTS, THF 65% 46b 3 TBDPS TBDPSCI, imidazole, THF 93% 46c In this case, it is now impossible to differentiate the two silylated groups of compound 46c with fluoride ions. So we had to find other conditions of deprotection which would be selective for t-butyldimethylsilyl in the presence t-butyldiphenylsilyl to obtain alcohol .17 and n ~. ~, has diol 48 (diagram 10) ~ OTBDPS I OTBDPS I ~~ OH
/ Deprotection ~ or - / /
Me0 \ I, ~ OTBDMS Me0 \ I OH MeO w I i ~ OH
46c 47 4g Figure 10 Several attempts have been made under acidic conditions to deprotect t-butyldimethylsilyl selectively as described in Table 2. In the presence of p-toluene sulfonic in methanol or in ethanol, diol 48 is the only product obtained (inputs 1 and 2). In presence of an acid resin (Dowex) or pyridinium p-toluenesulfonate in protic solvents, alcohol 47 is obtained in varying yields between 45% and 98%
(entries 3 to 5). The best reaction conditions are those which use the p-pyridinium toluenesulfonate in 95% ethanol (entry 4). He is very important not not continue the reaction for more than 16 h because otherwise we observe as reaction undesirable the migration of t-butyldiphenylsilyl to lead to a mixture of very alcohols with difficulty separable.
Table 2: Deprotection conditions for disilyl 46c.
Entry 'Reaction Conditions ~~ p ~ Product Yield ...... ... ...... ... ...... .._........_ _: ......... ... ..... ..
....... ° .. ......... ......_.....
1 APTS, MeOH, 10 mm 96/0 48 2 APTS, 95% EtOH, 10 min 100% 48 3 Dowex SOw-x8, MeOH, 15 min 45% 47 4 PPTS, EtOH 95%, 16 h 98% 47 PPTS, iPrOH, 50 ° C, 2 h 55% 47 The problem of reduction of the amide of Weinreb 37 encountered in the way long A (diagram 8) being adjusted by replacing it with ester 44 (diagram 9), we can continue the sequence. , ~ or ~ - -sous ~ vndre au, l3 keto ester 18 (diagram ll). In one first ter ~~ ps. we oxidized 1 "aicool 47 under Swern conditions''' to obtain aldehyde 49 in a quantitative yield. We then reacted this aldehyde with the anion of fluorophosphonate 2815, the same reaction conditions as those used previously in the long route A (diagram 6), to obtain the ester a, ~ unsaturated 50 of trans geometry exclusively in a 93% yield. Then reduce the ester formed to the help of aluminum and lithium hydride to obtain alcohol 51 quantitatively.
It is transformed into chloride 52 corresponding with hexachloroacetone and the triphénylphosphine22 in a yield of 99%. We move this chloride with sodium iodide and with the dianion of methyl acetoacetate and we obtain the ~ keto ester 53 in a yield of 65%. By treating the silyl ether of this compound with fluoride tetrabutylammonium we we obtain alcohol 54, which has already been obtained (diagram 7), in a yield of 77%. There we it remains only to transform this alcohol into a pivaloic ester in the same conditions that those used in the long track A (diagram 11).
~ OTBDPS I 'OTBDPS
S ~ / n-BuLi, THF, -78 ° C
OH 100% ~ IH (Et0) P (O) CH (F) COpEt 28 Me0 Me0 93%

OTBDPS ~ OTBDPS
LiAIH4, THF ~ ~ HCA, PPh ~ _ i F 0 ° C, 100% '~ ~ F THF, 99%
W
Me0 C02And Me0 OH

OTBDPS ~ OTBDPS
Nal, NaH, n-BuLi F MeC (O) CHzC02Me, ~ IF
Me0 \ CI THF, 65% Me0 ~ OMe ~ OH ~ OPiv TBAF, THF ~ ~ J ~ PivCl-2.6-lut b 77% 'IF CH ~ CI2, 97'k IF
Me0 ~ ~~ OMe Me0 '~ OMe 54 IO ~ IOI ~ 8 OO
Figure 11 We thus obtain the ~ 3 keto ester 18 which is the precursor of the reaction macrocyclization in an overall yield of 28% in 17 steps. This long track B constitutes a clear improvement compared to long track A whose overall yield was 3.8% and this in with the same number of steps.

1.3 Macrocyclization and Transannular Diels-Alder Reaction The key step in the synthesis of 14 / ~ fluorodigitoxigenin 7 (diagram 3) is the reaction of Transannular Diels-Alder. To achieve this reaction, you must first macrocyclize the, the ~
keto ester 18. So we reacted the ~ 3 keto ester 18 with the N O-bis (trimethylsilyl) acetamide (BSA) at reflux in tetrahydrofuran for form ether corresponding silylated enol. This enol ether is added slowly (14 h) silylated at a solution of the palladium catalyst, namely tetrakis (triphenylphosphine) palladium (0) which has previously reacts with bis (diphenylphosphino) propane at a concentration of 2x10'3M.
The high dilution is very important because if we add the enol ether silylated to a solution of more concentrated palladium catalyst, we can see the appearance of products of polymerization as well as decomposition products. We have also noticed that synthesizing ourselves tetrakis (triphenylphosphine) palladium (0) z3 instead to use that commercially available, reaction yields are higher (between 10 and 20%). So using these reaction conditions '' za, we get the thirteen macrocycle 22 members in an 88% yield (Figure 12), which is excellent for a reaction macrocyclization.
it is now necessary to introduce a double bond of geometry c ~. ~ in? e 22 pc macrocycle ~~ ar tee ~~~ sea TC diene which is necessary to carry out the Diels reaction-Alder transannulaire.
To do this, we used the reaction conditions used by Bartonzs, z6, We so we reacted this macrocycle with benzeneselenin anhydride and with triethylamine and we obtained a single product (by 1H NMR), namely triene 15 in one 91% yield (Figure 12).
The geometry of the cis double bond formed (relative to the macrocycle) has been determined by comparison of our results (1 H NMR) with those obtained by Luc Ouellet'i, 14 in the frame of the preparation of macrocycle 12 (diagram 1). In addition, the geometry of double bonds of triene 15 will be confirmed by obtaining a single reaction product (TSC ') when Transannular Diels-Alder reaction.

EO
Pd (PPh3) a, Ph2P (CH2) 3PPh ~ IF
BSA, THF, reflux, 2 * 10'3M
M add. slow (2 p.m.), 88% Me0 ..- 22 EO
(PhSeO) 20, Et3N, CH2C12 I \ F
91% I ~ /
MeO
Figure 12 So we submitted our macrocycle 15, which contains the geometry diene trans-cis (TC ~ and the dienophile of cis geometry (C ~, under the reaction conditions of Diels-Alder transannular (DATA). The conditions of the DATA reaction consist of heat the macrocycle 15 in toluene at a temperature of 165 ° C for 4 h in presence of triethylamine in a sealed tube. We thus obtain the tetracycle 16 with geometry junctions of cycles formed TSC exclusively dar ~ 5 a r ° nd ° ment 78 ° ~ ô (diagram 13). The Transannular Diels-Alder activity is therefore highly stereoselective because we let's perfectly control the four new centers created during the reaction.
EOEO
/ F Et3N, toluene, 165 ° C ~
w ~ W
4 h, IHF sealed tube Me0 ~ 78% Me0 Figure 13 This result is explained by looking at the transition state of the reaction of Diels-Alder transannulaire (DATA) presented in figure 15. We see that the alignment of orbital in the transition state 55 is perfect for making the DATA reaction. We pouri ns ~
also observe that there is no significant steric interaction which could prevent DATA's reaction to occur. The other possible DATA product would be tetracycle with stereochemistry at the cis-syn-trans cycle junctions (CST). In looking at the state of transition with a molecular model to get this tetracycle, we can note that the alignment of the orbitals, to reach this transition state, is impossible to obtain. We so let's get only one product is tetracycle 16 with stereochemistry at the junctions of TSC cycles.
F
Me0 O ~ ~~ COZMe pA ~ q H '=' F C02Me Me0 O
~ O

Figure 15: Transition state during the Diels-Alder reaction Transannular.
1.4 Chiral transannular Diels-Alder reaction 1.4.1 Results ant ,, err.e ~ adv Now that we have the racemic tetracycle 16, it would be interesting to try to do the same Diels-Alder transannular reaction but enantioselectively.
To do this, we are based on the results that have been obtained previously in our laboratory by Pascal Langlois2 '(figure 16). Indeed, this one demonstrated that both macrocycles of geometry TC'C 56a and 56b with alcohols / 3 in diene alpha (at the position C-17), go through the transition states 57a and 57b respectively. In the conditions of Diann-Alder transannular reaction, only one product is observed in each case, either TSC 58a and 58b tricycles with syn alcohols compared to methyl at the position C-13.

Me OTBDMS Me OTBDMS
. ,, E ~
ÖR I / OR
E = C02Me EE = C02Me ER = MOM E. R = MOM
E
56a racemic TBI 56b racemic TBI
H, -, .OR H% ,. OR
E
E H --- ~ OTBDMS EE OTBDMS
57a E 57b DATA DATA
Me OTBDMS Me OTBDMS
m 13 yE 13., VE
EH ÖR E ~~ OR
EE
58a TSC 58b TSC
figure 16: Pascal Langlois' previous results for I <b Diels reaction-Alder Transannular.
1.4.2 Synthesis of the macrocycle and formation of the tetracycle By reporting these results to our macrocycle, we have deduced that reducing the ketone to position C-17 of macrocycle 15 chirally, we can get the tetracycle 16 enantioselectively.
First, we reduced the macrocycle ketone 15 to alcohol in a way racemic to optimize the conditions of the Diels-Alder reaction transannular (diagram

14). Since we are in the presence of a ketone a, ~ unsaturated, the conditions of reduction developed by Luche28 were used. Under these conditions, the chloride cesium is a Lewis acid and it complexes with the ketone to promote reduction 1-2 of the ketones a, ~ unsaturated instead of the reduction 1-4. So we have treated on macrocycle 15 with cesium chloride heptahydrate followed by borohydride sodium and we obtained the macrocycle 59 in a yield of 88%.
EOE GOLD
NaBH ~. CeC13.7Hz0, MeOH, CHZCI2 (1: 1) Me0 ~ $ 8% Me0

15 59R = H
Protection 60 af R = groupings protectors Figure 14 To do the transannular Diels-Alder reaction (DATA), we need protect alcohol free from S9 macrocycle. Different protective groups have been used to 'check their influence on DATA's reaction. So we synthesized three ethers silylated from different sizes (inputs 1 to 3, compounds 60a, 60b and 60c), two ethers (entries 4 and 5, compounds 60d and 60e) and an ester (entry 6, compound 60 ~. All conditions reaction as well as the yields are described in table 3.

Table 3: Conditions and yields for the protection of macrocycle alcohol 59.
~ A Input y ~ M ~ M ~~ Y Reaction conditions ~~~~~~ »~~ sm. ~~ R ~~~ W ~~~~~~ Efficiency ~~ Product 1 TBDMSCI, imidazole, THF TBDMS 92% 60a 2 TBDPSCl, imidazole, THF TBDPS 47% 60b 3 TIPSOTf, 2,6-lutidine, CHZClzTIPS 54% 60c 4 MOMCI, DIPEA, CHZCl2 MOM 58% 60d EVE, PPTS, CHZC12 EE 100% 60th 6 PivCl, DMAP, pyridine Piv 100% 60f There are two transition states that are possible for Diels' reaction -Alder transannulaire on macrocycles 60a to 60f as illustrated in figure 17. The state of transition 61 is the more favored and it corresponds to that proposed by Pascal Langlois2 ~ in the figure 16. This last leads to Diels-Alder Transannular 63 product with junctions TSC cycles and the alcohol protected at position C-17 is found syn compared to the ester at position C-13.
F 1 ~ 0 Me0 OH ~ _ ~~ E _ _ -OR - 'Me0 O
61 af 62 af DATA ~ DAT ~ a.
OR _E OR

I, Fi FI j F
Me0 /
me0 63 af 64 af Figure 17: Transition states for the Diels-Alder reaction Transannular.
The transition state 62 is also possible (FIG. 17). However, we can see that there is a strong steric interaction between the diene proton and alcohol protected. This state of transition is therefore disadvantaged compared to the transition state 61. This second state of transition gives us the tetracycle 64 with the cycle junctions also TSC but with alcohol protected from position C-17 anti versus ester from position C-13.
All the macrocycles in Table 3 were therefore subject to the conditions of Diels' reaction Alder transannulaire. For the reaction to be complete, heat the macrocycles 60a at 60f in toluene in the presence of triethylamine at a temperature of 220 ° C in a tube sealed over a period of 16 h (diagram 15). We find that it takes a temperature higher as well as a longer reaction time for the DATA reaction either complete compared to the DATA conditions of macrocycle 15 (diagram 13), which has a ketone at position C-17 (165 ° C and 4 h). We can deduce that the ketone plays a role in decrease in energy of the transition state but no extensive study was made.
E OR E OR E OR
F
Et3N, toluene _ 220 ° C, 4 p.m. ~ \ 'J ~~ F ~ \ F
Me0 ~ Me0 ~ '~ / Me0 60 af 63 af 64 af Figure 15 The results of the transannular Diels-Alder reaction (DATA), macrocvcles 60a to 60f, are presented in Table 4. As the reactions were carried out on small quantities, yields are not all indicated. Plus the products trétracycliques obtained decompose quickly. The ratios were calculated from 1 H NMR spectra crude compounds.

at 8 Board 4:
ratios obtained for the reaction from DATA
sure the macrocycles 60a 60f.

Entre Macrocycle ~ R ~, ~ W ~ w ~~~ - ~ x ~ HR Efficiency Ratio (63 M w ~: 64) _1. _.... ~. ~ 0 ~. ........ _ .. TBDMS. ..... fi3, o / .... .. -.5.8 _........ ........... ........ _. l .............
._ 2 60b TBDPS not determined 1.8: 1 3 60c TIPS not determined 5.6: 1 4 60d MOM not determined 3.0 ~ 1 60th EE not determined dcomp.

6 60f Piv not determined 4.0: 1 We were surprised with the results since we expected have only one product of DATA as in the results previously observed by Pascal Langlois2 ~
(figure 16). We can see that depending on the protective group used, the ratios of tetracyclic products vary between 1.8: 1 and 5.8: 1. The most weak was obtained with t-butyldiphenylsilyl (entry 2) and the best ratio with t-butyldimethylsilyl (entry 1). This result is very surprising because, by increasing the size of the group protector we should have gotten a higher ratio of tetracycle 63b and the reverse has occurred. This phenomenon can be explained by the fact that the phenyls of grouping t-butyldiphenylsilyl can stack ~ with the aromatic ring A
macrocycle (~ l) b, This is afraid ~ 'fet to decrease the energy of the transition state 62b which is the least favored and therefore increase the formation of unwanted product 64b (figure 17). For which is other macrocycles, the ratios of tetracyclic products are all weaker than 5.8: 1 and in addition, there are several decomposition products which are observed in 1 H NMR spectra of the crude products (inputs 3 to 6).
So we continue to work on the two tetracycles 63a and 64a, who have the butyldimethylsilyl as a protective group and the best ratio of desired product (5.8 1). To prove that we really have the two tetracycles expected, according to the states of transition of figure 17, we separated them by flash chromatography.
Then, we subjected them to the same deprotection and oxidation conditions.
first time, we hydrolyzed the silyl ethers 63a and 64a with p- acid toluene 4.9 in methanol to give us the corresponding alcohols. We have then oxidized these alcohols with Dess-Martin periodin29 to lead to tetracycle 16 (diagram 16).
E OTBDMS ~ OTBDMS
H
(/ hi FI / F
Me0 vv Me0 63a 64a 1) APTS, MeOH, 75% 1) APTS, MeOH, 75%
2) Dess-Martin, CH2CIz, 100% ~ 2) Dess-Martin, CH2CI2, 100%
EOEO
_ H
HFIF _ Me0 ~ Me0

16 16 Figure 16 Since we are in a racemic series, the two products obtained are necessarily identical. This tëtracycle 16 has already been obtained during the Diels' reaction Transannular alder (DATA) with the 1st macrocycle 15 comprising the ketone (diagram 13).
So we did the chemical proof that the two tetracycles 63a and 64a go through well the two transition states that we proposed in Figure 1 ?.
1.4.3 Chiral reducers The results we observed for the DATA reaction on the macrocycle 60a protected in the form of t-butyldimethylsilyl are encouraging. We get a 5.8: 1 ratio in favor of the desired tetracycle 63a and the alcohols 63a and 64a obtained are separable. There we would therefore be possible to synthesize tetracycle 16 enantioselectively using a chiral reagent for the reduction of ketone from macrocycle 15 (diagram 17).

~ H
y IF ~
Chiral reducers \
Me0 (~ v Me0 Figure 17 There is a large selection of chiral reducers in the literature. However, in considering the other reducible functional groups of macrocycle 15, the ester and the conjugated diene with ketone, the choice of reducers is limited. Most reagents likely to us give good selectivity during the reduction reaction, depending on the results of the literature, are presented in Table 5.
Board 5:
reduction nantioslective macrocycle 15.

- Between ~ ~ %% - ~ Reducers ~ e. ~. .. ~ _ .., ~ .. Conditions ~~~% Compose % ~ ~ YS ~~ w ~~ '-'. X ~. of ~ reaction ~~~ q ~ ~~; (Yield) -_._.__._._ .. _ ~ lpine ~ Bor ......_..__ .._._..._ _....._.___.___ .. _. .
_.___;. .. ____ w p __._ _... _._ .. _____._ _.__..... __ .. decom.
1 R- donkey THF, ta 2 Binal-H THF, -78C ta 15 Ph 1. BH3.Me2S, THF, 0i. ' 1S

__ ~ Ph , N 2. BH3.Me2S, THF, 0 dcomp.
, 0 C ta H
'B

Me 65 4 H Pfph 1. BH3, THF, 0C, 2 h dcomp.

0 2. BH3, THF, 0C, 30 min 59 impure (11%) N, B

66 Me ~ NHPh 1. LiAlH4, Et20, -78C, 59 impure (20%) 5 a.m.

2. LiAlH4, THF, -78C, dcomp.
3 hrs sl First, we tried to do the enantioselective reduction with two commercially available reducers, namely the R-Alpine-Borane3 °
(entrance 1) and the Binal-H31 (entry 2). In the first case, decomposition products were observed and in the second case, the starting macrocycle was recovered. So we have synthesized two oxazaborolidines (compounds 65 and 66) which are compounds developed by Corey32 (inputs 3 and 4). These two compounds complexed with boron hydride are known in Literature to reduce α, 13-unsaturated ketones with good yields and good sélectivités33.
They have even been used in several total syntheses to induce chiralité34. In our case, the best alcoo159 yield we obtained is 11%
with oxazaborolidine 66 (entry 4). In addition to having a low yield, it has turned out that Ia reaction was not reproducible. So we synthesized pyrrolidine 6735 so enantioselective. Using the latter with aluminum hydride and lithium we let's get a chiral source of hydride. These reaction conditions are used in the literature to reduce unsaturated ketones to l3 and keto esters way enantioselective36, 3 ~. In our case, we got 20% return alcohol 59 (entry 5). Again, the yields are not reproducible and the purification of desired alcohol is very difficult.
3_.e ~ results obtained during the enantiosélevtive reduction reaction of macrocycle 15 have was not very encouraging. So we decided to continue, for the moment.
the development of the sequence up to 14 / .3 fluorodigitoxigenia 7 with the tetracycle 16 racemic.
In summary, three synthetic routes were used to synthesize the ~ -keto ester 18, precursor of the macrocyclization reaction. The shortest route, which is inspired by the way used in our laboratory for the synthesis of l4 ~ hydroxysteroids ~ l, a been abandoned quite early because Wittig's reaction to introduce fluorine gave us a mixture of olefins with a low yield (diagram 4). The other two tracks, which are more long we have allowed to synthesize the ~ 3-keto ester 18 targeted. The path chosen is that which goes through ester 44 (diagram 9, long path B) instead of the amide of Weinreb 37 (diagram 7, path long A), because the yields obtained throughout the sequence are much higher.
We then did the first key step in the sequence, the reaction of macrocyclisation on the ~ -keto ester 18. The macrocycle 15, with the double bonds of trans-cis-cis geometry (TCC), was therefore synthesized with very good yields. We we finally did the second key step in the sequence Diels' reaction Transannular Alder (DATA) on the TCC 15 macrocycle.
tetracycle 16 with stereochemistry at the junctions of cycles formed TSC as we had it predicted. So we get tetracycle 16 in twenty linear steps in a excellent overall yield of 17.5%.

Chapter 2 Approval at position C-17 In this second chapter, all attempts to certify the ketone in position C-17 tetracycle 16 and its derivatives to aldehyde 68, will be presented as well as the formation of methyl. in position C-13 (figure 18).
EO Me CHO
Fi F ~~ ~ ~ \ / ~~ H \ F
Me0 / Me0 'v Figure 18: Aldehyde approval at position C-17 and formation of methyl at position C-13.
2.1 Approval of carbonyl at position C-17 Many types of approval are known in the literature. ~ ïr_c c ~~ .., ~
~ Mus communes is the Wittig38 reaction on an aldehyde or a ketone. There are several examples of this type of reaction on the ketone at the C-17 position of steroids39, which is very encouraging for we.
In our case, we want to add a carbon at position C-17 to get an aldehyde (compound 68). With this aldehyde, we could then synthesize the butenolide which found in the 14,13 fluorodigitoxigenin 7 targeted or do different Wittig reactions to obtain analogues at position C-17 of 14 ~ fluorodigitoxigenin 7.

The first approval attempts were made on the product of the reaction of Diels-Alder transannulaire is the tetracycle 16. We reacted the chloride salt (methoxymethyl) triphenylphosphonium with lithium düsopropylamide (LDA) or with bis (trimethylsilyl) potassium amide (KHMDS) to obtain the phosphorane corresponding. We then added ketone 16 to this mixture and we we did the Wittig39 ° reaction with different numbers of equivalents of the phosphorane (1 to 10 equivalent) and at different temperatures (--78 ° C at reflux). In all cases we don't have never obtained the desired enol ether 69 (diagram 18). Ä temperatures of -78 ° C until room temperature, the starting product is recovered and when heating the reaction, only decomposition products are formed.
OMe [Ph3PCH20Me] + CI-, bases w. - ~ - w Me0 I ~ HF THF, -78 ° C at reflux Me0 I ~ H

bases = KHMDS or LDA
Figure 18 We thought that the problem of the previous Wittig reaction was caused by the presence of the ester at position C-13 on tetracycle 16. So we reduced the keto ester of this tetracycle using lithium borohydride and we got the diol 70 with a 90% yield. We have selectively protected primary alcohol from this diol with the t-butyldimethylsilane to give us the corresponding silyl ether. We have then oxidized the residual secondary alcohol of this intermediate with the perruthenate of tretrapropylammonium (TPAP) and 4-methylmorpholine N-oxide (NMO) 4 °
for us give ketone 71 (diagram 19). We treated this ketone in the same terms Wittig reaction than those used above. Unfortunately only of the decomposition has been observed. It turned out that this ketone 71 is unstable in solution and therefore that it breaks down before reacting.

EOH
1) TBDMSCI, Et3N
LiBHa: MeOH (1: 1), DMAP. DMF. 84%
,.
HF Et20, 90% ~ 2) TPAP, NMO, 4 M
Me0 ~ CH2CIz, 80%

[Ph3PCH20Mel + CI ', KHMDS
Decomposition THF, -78 ° C at ta 71 unstable Figure 19 We therefore synthesized ketone 75 (diagram 20), which does not contain olefin to the position C, 1-Ct2, hoping that it would be more stable than ketone 71 with the olefin. For this do we hydrogenated the tetracycle 16 olefin with the catalyst of palladium (0) on barium sulfate and in the presence of hydrogen to obtain the saturated ketone 72 in one quantitative yield. We then repeated the same conditions of reaction than those shown in Figure 19. We reduced the tetracycle 72 for us give the diol 73 in a 94% yield. We then made the alcohol monoprotechon primary of this compound using t-butyldimethylsilane chloride followed by alcohol oxidation secondary to give us the tetracycle 75 with good yields (diagram 20).
Saturated Ketone 75 is now stable and has been subjected to different reactions listed in Table 6. First, we have attempted the Wittig39 ° reaction with chloride salt (methoxymethyl) triphenylphosphonium with two different bases. Again, we got decomposition and US
have recovered part of the starting product (inputs 1 and 2). We have attempted a second Wlttlg39a reaction on ketone 75 but this time with the méthyltriphénylphosphorane to give us the corresponding exo-methylene but we recovered that the product of departure (entry 3). One possible explanation would be that ketone 75 becomes enolous easily and that the Wittig reagent can act as a base and thus prevent the reaction from produce.
EOEO
H2, 5% Pd / BaSO ~, LiBH4: MeOH (1: 1) EtOH abs., 100% I ~ ~ F THF, 94%
HF
me0 me0 OH
OH
TBDMSCI, Et3N
DMAP, DMF, 76%
HF
Me0 ~ Me0 9: 1 a./3 TPAP, NMO, 4A MS ~
CHzCl2, 95%
me0 Figure 20 Since we had no success with Wittig's reactions to this ketone 75.
we have tried other types of approval reactions. Initially time we we reacted ketone 75 with the reagent from Tosmic4 '. The latter allows to go from one ketone to a nitrite in one step. In addition, this reagent has been used to approve a ketone at position C-17 of several compounds that are similar to tetracycle 75. We so we treated ketone 75 with Tosmic's reagent but we didn't observed that decomposition products (entry 4).

Board :
terms of raction for approval of the ctone and of the alcohol 74.

mlEnter ~ Prod. ~~~ M ~~ Reagents ~ Solvent T (C) Product ~ start ~

1.. 75 ...... [Ph3PCH20Me] + Cl-, ....... .t .. ~. ~ 5- + dcomp.
... KHMDS THF refuX ...
...

2 75 [Ph3PCH20Me] + Cl-, THF ta reflux75 + dcomp.
n-BuLi 3 75 Ph3PCH3I, PhLi THF ta reflux75 4 75 Tosmic, t-BuOK DMSO ta decomp.

75 PhS (O) CH2C1, t-BuOK t-BuOH ta 75 + dcomp.

6 75 PhS (O) CHZCI, n-BuLi THF -78 -20 75 + dcomp.

7 75 (CH3) 3SCH3SO4, NaOH CH2Cl2 50 75 + dcomp.
50%

8 74 TsCI, Et3N, DMAP CHZC12 ta reflux74 9 74 MsCI, Et3N, DMAP CH2C12 ta reflux74 We then tried to synthesize epoxides from the ketone of tetracycle 75 with different reagents which are well known in the literature 42-a3. In in all cases, the starting material has been recovered as well as decomposition products (entries 5 to 7).
Not all attempts to register ketone 75 have worked. Through therefore, we tried to turn 74 alcohol into a good group leaving for then move it. So we treated tetracycle 74 alcohol with the chloride ~ osyle reentry 8) and with mesyl chloride (entry 91 but in both case. ~ e product of departure has been recovered.
So we tried a final approval reaction on ketone 75 with the reagent of Tebbe44 to obtain the corresponding exo-methylene (diagram 21). In one first time, we treated ketone 75 with ten equivalents of this reagent at 0 ° C
but no reaction never happened. By warming up this reaction to room temperature, the formation of a new product was observed. By analyzing the NMR ~ H spectrum of the latter, We have find that exo-methylene at position C-17 was present but that it there was a further signal in the olefin region. This additional signal correspond to eliminating fluorine to give compound 76 which has two olefins.
this is explained by the fact that Tebbe's reagent can act as Lewis acid and that he is coming complex with fluorine to eliminate it.
To try to obtain exo-methylene corresponding to ketone 75 exclusively we we did the same reaction but with only one equivalent of Tebbe's reagent.
We have then obtained the olefin 77 mainly from the elimination of fluorine, ketone departure and the diene compound 76. With this result, we concluded that the speed of fluorine elimination reaction is higher than the rate of formation of the exo-methylene at position C-17. Therefore, we cannot use this method for homologate carbon ketone 75 because of the sensitivity of fluorine in These conditions of reaction.
DMS
Tebbe (10 eq.) DMS THF, 0 ° C to t., A. ~

MS
75 ~ -Tebbe (1 eq.) ~~
THF, 0 ° C at ta + P ~ od. departure M
76 X = CH2 77 X = O
Figure 21 The results, which we obtained with Tebbe's reagent, have allowed to notice that fluorine is not stable in the presence of Lewis acids and that it is easily eliminated, this that we hadn't expected. We will see later that this sensitivity with acids from Lewis will cause us other problems in our streak. However, this sensitivity of Lewis acid fluorine can be used to successfully transform res impossible at the start of our work.
We have also attempted to certify the tetracycle 72 ketone obtained antérieuremtnt (diagram 19) and which contains the ester at position C-13 (diagram 22). The reaction results The approval of ketone 72 are presented in Table 7.
EOE ÇRs Homologetion ~
H Fv Me0 ~ Me0 I i HF

Figure 22 Board 7:
terms reaction for approval of the ctone 72.

\ "Mntre T ~ '. ~ .. ~,. ~ .. ~.-A. ~. ~, Ractifs .., ._ ~~ olvant., .. ...., T ~ -...- ....-Produced p ~ ~. ~ N ~ ....-., ..- .. ~ .. ~, '. (C), x. .,.
. ~ ..

__ .. i ._..._..._..... ~ .__._ [Ph3PCH20Me] + Cl-, ~ THF ~~ ~~~. ta ~ 72 + dcomp ~
KHMDS ~ ~ reflux 2 Ph3PCH3I, PhLi THF ta reflux72 + deomp.

; i T'osmic, t-BuOK DMSO ta dcomp.

4 Tosmic, BH3.DMS DMSO ta dcomp.

S (CH3) 3SCH3SO4, NaOH 50% CHZClz 50 dcomp.

6 PhS (O) CHZCI, t-BuOK t-BuOH ta 72 + dcomp.

7 PhS (O) CH2Cl, n-BuLi THF ta 72 + dcomp.

Initially, the same reaction conditions of Wittig39 with the chloride salt of (methoxymethyl) triphenylphosphonate were used and we got the product of departure and decomposition (inputs 1 and 2). We also used the Tosmic 'reagent ~ I
but this time, with two different bases: potassium t-butoxide (entry 3) and boron hydride complexed with dimethylsulfide (entry 4). In both case we we only got decomposition. Reagents to try to synthesize epoxides 42-a3 have also been used but without any encouraging results (entries 5 to 7).
2.2. Approval of carbonyl at position C-17 with reactions of coupling The different approval reactions carried out on the ketone at Ia position C-17 of tetracycles 71, 72, and 75 did not work. Therefore, it is necessary find another type of approval reaction than those used so far. We have therefore directed to the coupling reactions to approve the ketone at the position C-17 of different tetracycles.
First, we tried coupling reactions on the ketone 72. It is necessary first synthesize the triflic enol ether of the latter to be able to perform this type of reaction. We therefore treated ketone 72 with the isopropylamide of lithium (LDA) to make the corresponding enolate. We then treat this enolate with Comins4s reagent to give us the triflic enol ether 79 in a yield of 90% (diagram 23). This reaction confirms that the enolization of the ketone at position C-17 is easy to do.
It is for this reason that the Wittig reactions, used previously to try to certify ketone 72, did not work.
We now have to try the different reaction conditions of coupling on this triflic enol ether 79. So we made two attempts to reaction of carbonylation46 on this triflic enol ether with two catalysts different palladium.
In both cases, the starting material was recovered quantitatively. We so we have performed the coupling of the triflic enol ether 76 with the tetrakis (triphenylphosphine) palladium (0), crown ether 12-4 and lithium cyanide4 ~. To our great surprise after two days of agitation, we finally obtained the nitrile a, / 3-unsaturated 80 in one yield of 69% (Figure 23).

OE OTf 1) LDA, THF, -78 ° C to 0 ° C \
THIS
F 2), ~, 0 ° C at ta ~~, HF
- NTf Me0 N 2 Me0 72 90% 79 t_iCN, Pd (PPh3) to PdCl2 (PPh3) 2, K2COg 12-C-4, benz CO, MeOH, THF
69% or Pd (OAc) 2, PPh3, Et3N
CO, MeOH, DMF
CN
\, Starting product / H Fv me0 Figure 23 Finally, the homologation of a carbon of ketone 72 at position C-17 accomplished, it takes now get the nitrite at this position with a config ~ .iration, (i and the group n2ethyl has the position C-13 in place of the ester.
To do this, we started with the catalytic hydrogenation of the double binding of nitrite c ~ /. ~ unsaturated 80 with palladium (0) and hydrogen. This has us leads to a separable mixture of nitrite at 81a and nitrite ~ 381b in a ratio of 1: 2 respectively.
To obtain the maximum nitrite ~ 3 81b desired, we performed the epimerization of nitrite a81a48. So we formed nitrilate from nitrite a8la and then we treated him with 2,6-di-t-butyl-4-phenol (BHT). We obtained 67% of the nitrite, Q and 13% of nitrite has a mixture of 5: 1 (diagram 24).

E CN E ÇN E CN
\ H ~ .5% PdIC_ EtOH abs.
hi F v 96% I j H ~~ '\ HF
Me0 Me0 Me0 8 ~ 81 to 81 b 1: 2 (a: / ~ separable 1) LiNEt2, THF, 0 ° C
2) BHT, ta 67% ~ and 13% a Figure 24 Figure 19 shows the protonation face of nitrilate 82, which comes from nitrite 81a, to lead to nitrite ~ 381b in the majority. Kinetic protonation therefore takes place on the a side of the molecule despite the concave side of the C-D48 system. We come later that to have a protonation on the face a of the molecule, the nitrite is in very effect important.
E CN
E _ ..
C ~ N _ ~
F ___ ._ _ MeO ~ ~~
82 '81 b ROH
Figure 19: Leading face on nitrilate 82.
So we have the desired carbon configuration at position C-17 of intermediate 81b which is in the 14, ~ fluorodigitoxigenin 7. It is now necessary synthesize the methyl at the C-13 position of this intermediate. To do this, you must reduce the ester to this position in alcohol, make a good grouping starting from the latter and the move with a hydride source.

The first attempt to reduce the ester of compound 81b was made with hydride düsopropylaluminium (Dibal). Under these conditions, we obtained that products decomposition because nitrile can also react with Dibal for us give reactions unwanted. In addition, Dibal can also be a potential Lewis acid and Therefore, perhaps the fluoride elimination side reaction could happen.
So we have opted for lithium borohydride to reduce this ester and we have got alcohol 83 in a yield of 89%. We then treated this alcohol with the tosyl chloride to give us tosylate 84 in quantitative yield (Figure 25).
E CN
LiBHd: MeOH (1: TsCI, Et3N, DMAP
THF, 89% CH2CI2, 100%
HF
me0 81 b 83 OTs CN
NaBH4, DMSO ~ Decompostion ¿'~! -L F 80 ° C
Me0 ww Figure 25 To get the methyl at position C-17, you have to move this tosylate to using a source hydrides. In many cases, the displacement reaction of a tosylate is made with aluminum and lithium hydride49. In our case, given the presence of nitrite on compound 84, we preferred to use sodium borohydride. In processing the tosylate 84 with this source of hydrides, we obtained products from decomposition (diagram 25). These come from the side reactions of nitrite with the sodium borohydride. So we decided to reverse the order of the stages of reaction that is to say, synthesize the methyl group at position C-13 before to perform approval of the ketone at position C-17.
2.3 Formation of methyl at position C-13 and approval of the ketone at the position To obtain methyl at position C-13, we used the mixture of diols 73 already obtained (diagram 20) and we made the monotosylate of primal alcohols.
Two methods can be used with comparable yields. The first method consists of treat the mixture of diols 73 with tosyl chloride and after several days we get the mixture of tosylates 85 in a yield of 87% (diagram 26). Regarding of the second method °, we first form the tin ketal with dibutylstananne and on treats it with tosyl chloride to give the same mixture of tosylates 85 in one 87% yield (Figure 26). It is advantageous to go through the tin ketal because the reaction is much faster.
We must now move the tosylate group that we have formed to using a source of hydrides to obtain the corresponding methyl. To do this, we processes the mixture tosvlates 85 with sodium borohydride and you get 85% of the alcohol 87 and 10% of 'oxirane 86 (diagram 26). The formation of oxirane 8ti comes from tosylate primitive with the alcohol j3 which is present in the mixture of tosylates 85. This alcohol move intramolecular tosylate to form oxirane 86 and this reaction is faster than that of the displacement of the tosylate by a hydride. The formation of a cycle at four members is usually unfavorable. In our case, the reaction is intramolecular and the alignment between alcohol, Q and the leaving group is good. We obtain so the formation of oxirane 86 as a side product of the reaction.
We must now obtain ketone 14 to perform the same reaction of coupling performed previously on ketone 72 (diagram 23). To do this, we treat alcohol 87 with the tetrapropylammonium perruthenate (TPAP) and 4-methylmorpholine N oxide (NMO) 4 ° and ketone 14 is obtained in a yield of 87% (diagram 26).
TsCi, Et3N, DMAP
) H CI-12CI2, 87%) H
° u NaBH4, DMSO
1) Bu2Sn (O) 80 ° C, 85%
benz: DMF (3: 1) reflux Me0 73 2) TsCI, ta, 87% g5 9: 1 (a: / ~ 9: 1 (a: ~
O
Me OH Me O
T ~ AP, NMO
4 MS. CH ~ CH
87%
HF ~ H FV ~ \ HF
Me0 Me0 ~ Me0 86 (10%) 87 14 Figure 26 Ä t ~ starting from ketonE 14, nn synthesizes the triflic enol ether of this last a ~~~: c lea conditions previously developed on ketone 72 (diagram 23). So we treat ketone 14 with lithium dopropopropylamide (LDA) to make the corresponding enolate then we add the Comins4S reagent. To our surprise, we have not got enol ether triflic desired but only the starting material. To promote enolate formation, we added hexamethylphosphoramide (HMPA) and dimethylformamide But again, there was no reaction that occurred (Figure 27).

1) LDA, THF, -78 ° C to 0 ° C
Starting product Me ~ CI
~ NI NTf2> 0 ° C at ta Fi F 1) LDA, HMPA, THF: DMF (1: 1) Me0 -78 ° C at ta 14 Starting material CI if ~ N NTf2, ta Figure 27 So we tried to do some homologation reactions directly on the ketone 14 (Table 8). Again, whether with the chloride salt of (methoxymethyl) triphenylphosphonium39 ° (entry 1), the reagent Tosmic4 ~ (entry 2) or well has chloro sulfoxide43 (entry 3), we recovered that products of decomposition or the starting product. Finding that the results approval were the same as those obtained with the other tetracyeles (tables 6 and 7), we have decided to drop that type of reaction and go back to the training of enol ether triflic corresponding to ketone 14.
Table ~ ~ Reaction conditions gation a. cto ~ .ie for the homolo of 14, l ntre -, ~ w, ..., ~ .. ~, "~~ ~. Solvent ~. a .. ... ~. ~" -w ~ Product Reagents 1N '"~ .-. ._. .._ ..,., - m. ~. ~, ~ T ~ (oCj., ~
.. ~. ~, ..

_.i ... ..._ - [Ph3PCH20Me ___THF_ .. _._ ta _ 14 ~ + .. a ~~ p ] + Ch, KHMDS ___... _. . __ reflux ___ . .

2 Tosmic, BH3.DMS DMSO ta dcomp.

3 PhS (O) CHZCI, n-BuLi THF -78 -20 dcomp.

It seems surprising that we cannot make ketone enolate 14 with the LDA
(Figure 27). Therefore, we deduced that the ketone enolate was probably formed but that the latter was too stable to react with the reagent of Comins45. This result is explained by the fact that lithium forms a bond with covalent character with oxygen and therefore the reactivity of the enolate formed is decreased. In taking in considering this reasoning, we decided to change the base used previously (LDA) to make potassium enolate instead of lithium enolate. We so we have treated ketone 14 with potassium bis (trimethylsilyl) amide (KHMDS) then with the Comins45 reagent and we got triflic enol ether 88 in a yield of 84% (Figure 28). We can now do the coupling reaction on this enol ether triflic. So we treated triflic enol ether 88 in the same conditions of reaction than those used previously 4 ~ to make the nitrile (Pd (PPh3) 4, LiCN, 12-C-4) and we obtained nitrite a ,, unsaturated 89 in a yield of 79%.
The olefin of this compound is then hydrogenated and 95% yield of a mixture is obtained separable from nitriles ~ 90b and a 90a in a ratio of 1.6: 1 respectively. We have tried to do epimerization of nitrite has minority with several different bases We have obtained only from decomposition products (diagram 28).
Me O Me OTf 1) KHMDS, THF, 0 ° C at ta \
LiCN, Pd (PPh CI ~ ~ = 12-C-4, benzene N z Me0 f ~ HF 2) ~ i NTf, ta Me0 I ~ HF ~ 9%
14 84% 88 Me CN Me ÇN Me CN
Hz, 5% Pd / C. ~
HF EtOH abs. ~ ~ FI \ HF
Me0 ~ ~ 5% Me0 me0 89 90a 90b 1: 1.6 (a: p) separable Bases = LiNEtz 1) Bases, THF, 0 ° C
LDA 2) BHT, ta KHMDS decomposition Figure 28 The structure of the majority nitrile 90b was confirmed by diffraction of the X-rays thus demonstrating unequivocally the stereochemistry at the junctions of the BCD cycles and the nitrile stereochemistry ~ at position C-17 (Figure 20).
Figure 20: X-ray structure of nitrile 90b.
2.4 Phenolic compounds It would be interesting to assess the biological activity of steroids which have a cycle A
aromatic and fluorine at position C-14. So we could compare these results biological with those obtained for 14 ~ -fluorodigitoxigenin 7 which we want to ynthétiser.
To obtain ring A in the form of phenol, the hydrolysis of methyl ether of compound 90b. So we treated this compound with boron tribromide and US
we obtained the corresponding phenol. We then protected this phenol with the t-butyldimethylsilane chloride to obtain compound 91 with 76% of yield for the two stages (diagram 29). The 1 H NMR spectrum of this compound:; or perfectly at product we are waiting for. However, when we analyzed the spectrum mass of this last we got, to our surprise, a higher mass corresponding to the substitution of fluorine at position C-14 with bromine. Boron therefore comes complex fluorine to give us the tertiary carbocation and instead of having elimination of a proton (like in the case of Tebbe's reagent), there is a bromide which attacks this last for lead to compound 88. It is therefore a substitution reaction of SN1 type that is happening.
We have deduced that the bromide attacks by the least congested face of the molecule either the face / j (convex face). We thus obtain the CD cis cycle junction only. This result gives us another example of the sensitivity of fluorine to Lewis acids.
We also did the same reaction with boron trichloride to check the generality of the reaction. We therefore treated compound 90b with trichloride boron followed by t-butyldimethylsilane chloride and we got chloride 92 correspondent in a 84% yield for the two stages.
A methyl ether is very difficult to hydrolyze and several conditions in the litterature use a Lewis acid to do the reaction5 ~. Therefore, fluorine at position C-14 of tetracycle 90b is incompatible with these reaction conditions. We have so left of rated the synthesis of phenolic compounds since our main objective is the synthesis of l4 ~ fluorodigitoxigenin 7.
Me CN Me CN
1) BX3, CH2CI2 r 2) TBDMSCI, imidazole ~
IH ~ F THF ~. ~ H ~ X
MeO ~~, _ ~; f. ~: ASO ~
90b 91 X = Br X = Br Yield = 76%, 2 steps 92 X = CI
X = CI Yield = 84%, 2 steps Figure 29 In summary, we carried out several types of approval reaction in this chapter on several different compounds. It is ultimately the coupling reaction to palladium with the lithium cyanide which allowed us to obtain the nitrite / 3 at the position C-17 desired. We we also synthesized methyl at position C-13 which is found in the 14 ~
fluorodigitoxigenin 7 targeted.

We found that fluorine at position 143 of the tetracycles synthesized is very sensitive to Lewis acids and can react elimination or substitution with these. Knowing this will require more attention under the conditions of reaction that we will use in the rest of the sequence. On the other hand, as we will see later, we will be able to take advantage of these side reactions that We have observed. These will allow us to carry out a transformation very chemical interesting and not planned at the start of our work.

Chapter 3 Functionalization of cycle A
In this chapter, we will discuss the functionalization of cycle A to from compound 87.
The plan of the synthesis is presented in diagram 30. To begin the sequence we want to do a Birch reduction of compound 87 followed by hydrolysis ether of methyl enol formed which will lead us to the ketone ~, unsaturated therein corresponding. he then protect the alcohol at position C-17 to obtain the compound 93. Ketone at the position C-3 would then be reduced to obtain alcohol / 3. We have planned that this alcohol will direct the cyclopropanation reaction on the face of the dessuss2 and thus, we can get cyclopropane 94. There would then be oxidation of the alcohol at position C-3 of this intermediate in ketone. The latter will be used to open the cyclopropane for us lead to methyl at position C-10. Then you would have to hydrogenate the compound olefin 95 which will allow us to obtain the junction of the AB cis cycles. Finally, it will take reduce the ketone to have alcohol, Ci 96 and functionalization of cycle A
will be finished.
Me ~ H Me ~ MOM Me OMOM
i, _- ....
1 Birch --------- ~ 1) Reduction _-_-_ 2) Hydrolysis ~ 2) Cyclopropanation Fi F 3) Protection ~ hi F
MeO ~ ~ O HOJ ~

Me ~ MOM
Me ~ MOM
1) Oxidation ~ Me 1 _H dro ena_ti_o_n Me 2) Opening _ 2) Reduction F! ~~~ F
O
HO

Figure 30 3.1 Synthesis of the model for cyclopropanation To introduce methyl at position C-10 of 143 fluorodigitoxigenin 7, we want go through a cyclopropanation reaction. To study this reaction of cyclopropanation, we have synthesized a model from 3-O-methylestrone 97 which is available commercially.
First, you have to do the Birch reduction of 3-O-metylestrone 97 for give us the corresponding methyl enol ether followed by the hydrolysis of the latter for lead us to the ketone, l3, ~ unsaturated 98. These two stages are known in literature and we obtained a comparable return53 (93% instead of 99%). Then we have protected the alcohol of compound 98 using methoxymethyl ether chloride and dopropopropylethylamine to give us compound 99 in a 78% yield (diagram 31). Here, you have to be very careful because this compound is very sensitive in acidic conditions. In these conditions, we observe the migration of the double bond to give us the ketone conjugate corresponding to compound 99. So always stay in the middle basic (extraction and flash chromatography) when handling this ketone dd, y unsaturated.
But then reduces the ketone of this interrr ~ ediary 99 with hydride of tri (F ~ ~ butoxydel aluminum with ec of lithium to lead us to an inseparable mixture alcohols a and, G- ~ 100 in a ratio of 6: 1 respectively (there is only the 100 isomer which is represented in figure 31). It's the best ratio we've got and it's comparable to results obtained on similar tetracycles in the literature 4. With this mixture alcohol, we did two cyclopropanation tests. In the first case we have used an amalgam of zinc-copper and duodomethane (cyclopropanation of type Simmons-Smith55) and we obtained a mixture of starting spirits and cyelopropanes 101.
In the second case, we reacted the 100 alcohol mixture with the diethyl zinc and with düodométhane56. We got a quantitative return of one mixed inseparable from cyclopropanes a and ~ 101 in a ratio of 6: 1 respectively (diagram 31).
We can therefore see that the cyclopropanation reaction with the diethyl zinc is very effective. Additionally, alcohol at the C-3 position of the steroid directs the face perfectly of attack during the cyclopropanation reaction since we obtain the same ratio diastereomeric starting alcohols 100 and cyclopropanes formed 10a.
O Me Me 1) Li, NH3, EtOH, THF OH
-78 ° C to -30 ° C MOMCI, DIPEA
HH 2) 50% HOAC / H20, THF (~~.-H ~ ~ CHpCl2, 78% ~.
°, ~ .1, ~~.
MeO ~ ~ 93 / o, 2 steps p 97- gg Me OMOM Me OMOM
LiAI (Ot Bu) 3 THF, 0 f..1 H 91 O HO ,.
99 100 6: 1 (a: ~
Zn (Cu), CH21 Zn (Et) 2, CH212 CH2Cl2 Et20 100%
Me OMOM Me ~ OMOM
, _ ~ ~, .., i -i- Starting product \\. ~) '', "_ H ~ H
HO HO J ~ j ~
101 6: 1 (a., T ~ 101 6: 1 (a; ~
Figure 31 3.2 Cyclopropanation reaction with the mixture of alcohols 106 3.2.1 Synthesis of alcohol / B
The encouraging results we have obtained with the model 97 compound to test the cyclopropanation reaction (diagram 31) allow us to attack the synthesis of our own molecule in a positive way. To start the sequence, we did reduction of Birchs ~ of alcohol 87, already obtained before (chapter 2, diagram 26), with:
lithium metallic in a mixture of tetrahydrofuran, liquid ammonia and ethanol for lead to the corresponding methyl enol ether. We then treated this intermediate with a 1: 1 mixture of 50% acetic acid in water and tetrahydrofuran to give us the ketone ~ 3, unsaturated there 102. We protected the alcohol of this intermediate with methoxymethyl ether chloride to give us the compound 93 (diagram 32). Like the model 99 compound, this intermediate is very sensitive in the middle acid because the double bond migrates easily to conjugate with the ketone which is at position C-3. We must therefore handle this ketone, l3, y ~ unsaturated 102 with precautions.
Me ~ H Me OH
1) Li, NH3, EtOH, THF
r ,. ,,. ,, '__ -78 ° C to -30 ° C, ~ f ~ ~ _MOMCI, DIPEA
-2150% HOAc / H ~ O, iHF H ~~ F CH2CI
me0 O

Me OMOM Me, OMOM
LiAl (Of-Bu) THF, O ~ C
63%, 4 steps ~ HF
O HO ,.
.93 103 6: 1 (cr., 131 Figure 32 5%
To continue the sequence, the ketone of compound 93 is reduced using sorting hydride (t-butoxide) of aluminum and lithium and an inseparable mixture of alcohols a and ~ 3 103 in a ratio of 6: 1 respectively. Only the a 103 isomer is shown in the diagram 32. We thus obtain a product which is stable in a yield of 63%
for the four final steps. This return is comparable to that which we obtained for the series model which was 66% for the same four stages (Figure 21).
Gum demonstrated in the model series, alcohol at position C-3 of the tetracycle 100 runs the attack face during the cyclopropanation reaction (diagram 31). It is necessary so reverse the stereochemistry of alcohol 103 to obtain the cyclopropane of configuration / j.
To do this, we used the Mitsunobus8 reaction.
First, we treated the 103 alcohol mixture with acid benzoic acid, triphenylphosphine and with diethyl azodicarbonylate (DEAD). We we obtained the benzoic esters 104 and the diene 105 in a 1: 1 mixture who will be separated in the next step (Figure 33). This side elimination reaction is explained by the fact that we have a double bond at position / 3, y alcohols 103.
This double bond has the effect of making the protons in a of the latter more acidic. We so we have a competition between the substitution reaction and the elimination reaction of 3nélar. ~ Age of alcohols 103 at position C-3. Depending on the result we got, to say a 1: 1 mixture of substitution product 104 and elimination product 105, speeds of each of these two reactions are similar.
To carry out the cyclopropanation reaction, the alcohol must be released at the position C-3 which is as a benzoic ester. To do this, the mixture of esters 104 is treated and diene 105 with aluminum and lithium hydride and the mixture of alcohols is obtained 106 in a ratio of 6: 1 in favor of alcohol ~ 3dired. We can now separate the diene, trained in step above, a mixture of alcohols. This gives the mixture of alcohols 106 in one 50% yield for the last two stages. There is only the isomer /.3 which is shown in figure 33.

Me ~ MOM Me OMOM Me ~ OMOM
PhCO ~ H, PPh ~ _ h1 F DEAD, THF IHF
h1 F
HO ~ '~~ PhC02 103 6: 1 (a. ~ 104 6: 1 (/ 3: a) 1 OS
LiAIH4, THF, 0 ° C
50%, 2 steps Me .OMOM
HF
HO
106 6: 1 (~ i: a) Figure 33 3.2.2 Cyclopropanation reaction We now have the alcohol mix .106 which contains the alcohol / 3 majoa-itairement (6: 1). So we're ready to do the cyclopropanation reaction on this mixture with the method developed on the model. All tests of cyclopropanation on the mixture of alcohols 106 are presented in table 9 and the reaction is illustrated in the figure 34.

Me; .OMOM Me, OMOM
Cyclop ~ opanation Fi F ~ HF
HO HO
106 6: 1 ((3: a) 94 6: 1 ((3: a) Figure 34 First, we took over the conditions of cyclopropanation used on the compound model 97 (diagram 31). So we treated the mixture of alcohols 106 with ten equivalents of a 1 M solution of diethyl zinc in hexane and with ten equivalents of düodométhane. These conditions, which gave us a quantitative return of cyclopropanes 98 with the compound compound 97 (scheme 31), gave us traces of cyclopropanes 94 and the mixture of starting alcohols 106 (entry 1). We so we have increased the reaction time and we got an inseparable mixture of the cyclopropanes 94 and starting alcohols 106 in a 1: 1 ratio (input 2). By heating the reaction mixture at reflux to increase the rate of the reaction cyclopropanation, we only got decomposition products (entry 3). We have so do it again reaction at room temperature over a longer period of time and with a plus large excess of reaction. ~ We: frost then obtained 1f: product cie departure from majority cat way a trace of the cyclopropanes 94. The cyclopropanation reaction was therefore not not reproducible with diethyl zinc in solution. To check the quality of responsive we have repeat the cyclopropanation reaction on the model 97 compound (diagram 31) and We have obtained a mixture of starting material and cyclopropanes. Therefore, the diethyl zinc solution was of poor quality due to its instability over time. We so we did cyclopropanation tests with pure diethyl zinc.
Diethyl pure zinc is a flammable product in the presence of oxygen. It is necessary so do very be careful when handling it. We treated the mixture of alcohols 106 with ten equivalents of pure diethyl zinc and ten equivalents of duodomethane (entry 4). Those are the same reaction conditions which allowed us to obtain the 1: 1 mixture of cyclopropane and starting material (entry 2). These reaction conditions have leads to products of decomposition that come from the fluorine elimination reaction to the position C-14 (entry 4). During cyclopropanation reactions with diethyl zinc to solution we have not observed the presence of such side reactions. The diethyl pure zinc is so much more reactive and you have to be very careful because it can act as acid of Lewis and thus lead to the elimination of fluorine. To delete this side reaction elimination, we did the reaction at -10 ° C and we got a mix inseparable from starting materials and cyclopropanes 94 in a ratio of 2: 1 respectively (input S). So we did the reaction in the same conditions but at 0 ° C and we got the starting materials and the decomposition (entry 6). We then reduced the number of equivalent of diethyl zinc but the results were not more encouraging (entry 7).
Board :
testing of cyclopropanation sure the mix alcohols 106.

~ .ntre ~ - ~~~ Reagents ~ - ~ ......, .....- ~ Solvent., ... aMT .. (C ~ ~~. ~~ Product ~. ~, r .. TempsF (h) ~ x ~~

1 ...___._ l ~ .q: ZnEt2_11VI / hex, .l ~ .eqCHZC ~ Z ... .ta 4 ._106 - + _ trace _CHZIZ .... _ 94 2 10 q. ZnEtz 1M / hex, 10 CHZCIz ta 16 106: 94, q. CH2h 1: 1 1 10 q ZnEt2 1M / hex, 10 CH-Z, Ch refla.x: 16 dcomp.
q. CHZI2 ~ t 10 q. ZnEtZ, 10 q. CH2I2CH ~ C12 ta 5 decomp.

10 q. ZnEt2, 10 q. CHZIZCHZCIZ -10 3 106: 94, 2: 1 6 10 q. ZnEt2, 10 q. CH2I2CHZCh 0 6 106 + dcomp.

7 2 q. ZnEt2, 2 q. CHZIZ CHZCIZ ta 3 106 + dcomp.

8 10 q. ZnEt2, 10 q. CH2I2CH ~ C12 -10 16 dcomp.

9 5 q. ZnEt ~, 5 q. CH2I2 Et20: hex ta 5 106 5 q. ZnEt2, S q. CHZIZ CHZCI2: hext.a 1 106 + dcomp.

11 10 q. ZnEtz, 20 q. CHZIZCHZCh ta 1 106 + dcomp.

12 10 q. ZnEtz, 20 q. C1CHZICl (CHZ) ZClt.a 1 decomp.

13 Zn (Cu), 10 q. CHZIZ Et20 to 3 decomp.

In tests 4 to 7, we changed the reaction temperature as well that the number of diethyl zinc equivalents. We now have to change the time of the reaction.
We therefore treated the mixture of alcohols 106 at -10 ° C (temperature optimized) on a 16 h period with ten equivalents of pure diethyl zinc and we got only decomposition products (entry 8).
Throughout the tests, we kept dichloromethane as solvent because it is the no longer used for this type of reaction59. In addition, the reaction of cyclopropanation is a lot slower and it requires a higher temperature when it is performed in solvents such as ether and tetrahydrofuran. We still did some tests by changing the reaction solvent or by mixing two solvents. In all cases, starting materials and decomposition products have been recovered. (entries 9 to 11).
So we changed the düodométhane for chloroiodométhane because it is used in some examples from the literature b ° but we only got the decomposition (entry 12). We have tested the reaction conditions of Simmons-Smith55 with the zinc-copper amalgam and again we only got the decomposition (entry 13) In summary, the best result obtained for the cyclopropanation reaction was with the diethyl zinc solution but we could not repeat this result (entry 2). With the diethyl pure zinc we got a 2: 1 mixture of the starting materials and of cyclopropanes. In all other cases, we obtained a mixture of products of departure and decomposition products.
3.3 Alternative method of introducing methyl at position C-10 Another method to introduce methyl at the C-10 position of steroids consists of synthesize an epoxide a with the double bond at the junction of the AB rings and to open this Ö ~
epoxide with a source of CH3-. There are few examples of this type of reaction in the littérature6l. So we decided to test a compound model to test the reaction.
To do this, we used intermediate 107 (intermediate product from the reduction of Birch (diagram 32)) and we treated it with 1,2-bis (trimethylsilyloxy) ethane and triflate trimethylsilyl. We then added the pyridine to the mixture reactive to destroy traces of acid and we obtained ketal 108 in a yield of 56%
(diagram 35).
Under the reaction conditions, the alcohol at position C-17 was protected in the form of trimethylsilyl. This compound was then treated with m- acid chloroperoxybenzoic and we we obtained an inseparable mixture of 109 a: / 3 epoxides in a ratio of 2.5 : 1 respectively. We then tried to open this mixture of epoxides to the help of methyllithium or methyl bromide magnesium but in both cases we have only obtained the starting product. By adding cerium chloride with the methyllithium, we have seen the appearance of decomposition products (diagram 35).
Me OH Me OTMS
1) TMSO (CH2) 20TM I \ S
TMSOTf, CHzCl2, -78 ° Ç ~ mCPBA, CH ~ Ch_ H ~ Hv 2) pYr, ta ~ (H, H 88%
MeO ~ - 56%% O ~~ -_. I ~
107 ~ ~~ 'O

Me OTMS MeLi THF: DMF (1 ~) Starting material OO h1 hi MeMaBr ~ Starting material + decomposition THF or DMF
reflux 109 2.5: 1 (a. ~ CeCI ~, MeLi ~ Starting material THF
Figure 35 The methyl at position C-10 could therefore not be introduced by this method. Moreover, the different cyclopropanation tests to introduce this methyl did not given none encouraging result. To continue exploring the sequence, we so we decided to functionalize cycle A with a proton at position C-10 instead of the methyl.
3.4 Functionalization of cycle A with the proton at position C-10 To complete the functionalization of cycle A with a proton at position C-10, it takes everything first repeat the Birch reduction on alcohol 87 to obtain the ketone a, ~ unsaturated corresponding. Then we have to reduce the olefin of this intermediate to lead to the join the AB cis rings and reduce the ketone to position C-3 to get alcohol from stereochemistry ~.
So we redid the Birch reduction on alcohol 87 to get enol ether corresponding methyl. We treated this intermediate with acid hydrochloric and we got a mix of two products. The first product is ketone a "Q-unsaturated 110 desired and the other is the corresponding product with a proton to instead of fluorine at position C-14. The formation of this secondary product allowed us to find that the fluorine is sensitive to the conditions of radical reduction. By center, this result is very surprising because we had not observed the formation of this product in the ~ R ~ th Birch reduction conditions previously used (Figure 32). Through therefore, we deduced that the conditions for the Birch reaction were not reproducible and that it had to rework them. So we redid the Birch reduction of alcohol 87 at - 'i8 ° C at instead of -30 ° C and over a period of 2 hours instead of 5 hours. In these modified conditions, we we obtained the ketone a, ~ 3 unsaturated 110 desired in a yield of 77% and We have eliminated the formation of the secondary product (Figure 36).

Oh me Me _ 1) Li, NH3, EtOH, THF OH
-78 ° C to -30 ° CH ~ H ~, 5% Pd / C ~
I ~~ F 2) HCI 1N., THF ~ / HF ACOEt, 60%
Me0 77% O

OH O
My me TPAP, NMO
H ~ '4 ~ MS. CH ~ CI ° _ f. ~ ~ L-Sélectride w gg% ~~ THF, 100%
i- ~
HFHF
OO
111 + 27% of 112 (notches) 113 Me O Me OTBDMS Me O
H ~ TBDMSOTf _ H \
Et3N, CH2CI2 + n H
H ~ FHF ~ HF
HO TBDMSO ~ TBDMSO

TBAF, THF
_ _. ._ .. _ g2%, 2 stages __ ... . _ Figure 36 The problem of reduction of Birch being solved, we hydrogenated the olefin ketone a, ~ -unsaturated 110. We obtained 6U% yield of ketone 111 with the junction desired AB cis cycles and 27% yield of ketone 112 with the joining cycles AB trans (diagram 36). We still have to synthesize alcohol / d at position C-3 of this compound.
Compound 112 is therefore oxidized with tetrapropylammonium perruthenate and with the N
4-methylmorpholine 4 ° oxide to give us diketone 113 in a yield quantitative. Now reduce the ketone to position C-3 selectively in the presence of the one at position C-17. To do this, we use a reducer which is sensitive to steric hindrance is L-Selectride62 because the ketone at position C-

17 is more congested due to the quaternary center which is in a of the latter. We treat so the diketone 113 with this reducer and we obtain alcohol 114 ~ 3 at position C-3 as single product in a quantitative yield. It is by referring to similar steroids (with the junction AB cis) which have been reduced with the same reducer as we have deduct the alcohol stereochemistry at position C-36z-63. pn then protects this alcohol 114 with the t-butyldimethylsilane triflate and silylated ether 116 is obtained as well as silylated enol ether 115 as a mixture. This mixture is treated with fluoride tetrabutylammonium (TBAF). Under these conditions, only the silylated enol ether reacts with ions fluorides. Compound 116 is therefore obtained in a yield of 92% for the of them final steps.
In conclusion, Birch reduction of the aromatic cycle worked well but the reaction cyclopropanation did not allow us to introduce methyl at the position C-10. The best result was an inseparable mixture of starting olefins 106 and of cyclopropanes 94 in a 1: 1 ratio. In addition, it was found that the reaction of cyclopropanation was not reproducible. All other attempts to optimize conditions of this reaction did not give the expected results. this is probably due to the sensitivity of fluorine; position C "~ 14 fa. ~ ea 5x conditions used to get the cyclopropane.
So we decided to continue the sequence with the proton at the position C-10 instead methyl and we got ketone 116 with very good yields. The news target is therefore 10-nos-14, Q-fluorodigitoxigenin 117 which is represented at Figure 22.

H

Figure 22; Structure of 1a10-nor-14 ~ -fluorodigitoxigenin 117.

Chapter 4 Combination of the results obtained for the functionalization of cycle A
and for approval at position C-17 This chapter describes how we combined the approval of the ketone position C-17 described in chapter 2 and the functionalization of cycle A
described in chapter 3.
These two model studies work very well separately. So you have to apply in the good order to obtain the targeted compounds.
4.1 Approval of the ketone at position C-17 with cycle A
functionalized The simplest method, to obtain aldehyde 118, is to make the approval of the ketone on compound 116 with ring A already functionalized (Figure 23). AT
from this aldehyde we could synthesize butenolide as well as analogs 119 at the position C-17 of 10-nor-14/3 fluorodigitoxigenin 117 in just a few steps.
R
Me O Me ~ HO
HH ~~~ 1 / ~ ~ '~, - ~ i - ..
_.
T8ÜMS0 '~ TBDMSO

i H

Figure 23: Plan A for the synthesis of 10-nor-14, ~ fluorodigitoxigenin 117 and its analogous to position C-17.

To make the triflic enol ether on the model compound (with cycle A
aromatic described in diagram 28 in chapter 2), we used as a basis the bis (trimethylsilyl) potassium amide (KHMDS) followed by the reagent of Comins45 as source of triflate. By treating ketone 116 under the same conditions of reaction, us.
only obtained the starting product and decomposition products (Figure 37).
Me ~ Me OTf 1) KHMDS, THF, 0 ° C at ta _ l ~ ' CI ~~ H \
HF 2) ~ N ~ NTf2 ~ ta HF
TBDMSO TBDMSO

Figure 37 So we tried several other conditions to synthesize ether triflic enol 120 which are presented in Table 10. First, we have changed the source of triflate to be sure it was not the cause of the problem.
Unfortunately with the N phenyltrifluoromethanesulfonimide which is available commercially we we got the starting ketone and a lot of decomposition products (entry 2). We : wons dUllC tried to make enolate with bis (trimethylsilyl jamiclvire ae sodium (NaHMDS j and lithium dopropopropamide (LDA) with or without hexamethylphosphoramide (HMPA j.
In all cases, we obtained either the starting product or products of decomposition (entries 3 to 5).
So we used thermodynamic conditions to try to synthesize ether of triflic enol 120. A condition that is used frequently in the literature64, consists reacting a ketone with a congested base and with anhydride triflic like source of triflate. We therefore treated ketone 116 with a base called "
proton sponge "and with triflic anhydride but we only got decomposition (entry 6).
We changed the cluttered base to triethylamine and there was no no change (entry 7). At this point we realized that our ketone from departure was not 8 ~
stable in the presence of triflic anhydride. So we changed the source from trifla e for the Comins45 reagent and N-phenyltrifluoromethanesulfonimide. By treating ketone 116 with either of these reactants in dichloromethane or in tetrahydrofuran to reflux, we recovered only the starting product (entries 8 and 9).
Board :
terms reaction for synthtiser ther of nol triflic 120.

~~ Enter ~~ Reaction conditions. .. ~ p T ~ Solvent "~ F ~~~ Product ~ ~ '.... ~ _.nn ~~ Cj ....
mP ..

.1 ____. _._..._ l ~ -KHMDS- _2 ~ _. O ~ .. ~ _a.THF._ _ _1.16 or _ Convins. _ .. _ dcomp.

2 1) KHMDS 2) PhNTf2 0 ta THF 116 or dcomp.

3 1) NaHMDS 2) PhNTf2 0 ta THF 116 or dcomp.

4 1) LDA 2) Convins 0 ta THF 116 5 1) LDA, HMPA 2) Convins0 ta THF 116 6 Proton sponge, Tf20 ta CH2C12 dcomp.

7 Et3N, Tf20 ta CH2C12 dcomp.

8 Et3N, Convins ta reluxCHZC12 116 9 Et3N, DMAP, PhNTf2 reflux THF 116 The different reaction conditions that we tried to make enol ether triflic from ketone 116 did not work. According to the results We have got we think the triflic enol ether 120 is formed in some terms (inputs 1 to 3) but that it is unstable. It is for this reason that we do not are not able to isolate it and that we often get decomposition.
So we used other types of approval reactions from ketone 116 which would allow us to obtain aldehyde 118 (figure 23). The conditions of reaction and the The results obtained are described in Table 11. First, we have used several different Wittig39 reaction conditions to make the approval of the ketone 116. In all cases, only the starting material was recovered (entries 1 to 3).
We then reacted ketone 116 with the reagent from Tosmic4l with two solvents different. Whether with dimethoxyethane (DME) or with dimethylsulfoxide (DMSO), we got the starting product and products from decomposition (inputs 4 and 5). We treated ketone 116 with diethyl cyanophosphonate and with the lithium cyanide. These reaction conditions make it possible to pass from a ketone to a nitrite in a single step 65. In our case, we got that ketone from departure (entry 6).
Finally, we tried several different conditions to make époxydes4z-43.
In all cases, we obtained only decomposition products (entries 7 to 9).
Table 11: Reaction conditions for the approval of ketone 116.
Between Reaction Conditions Solvent T (C) Product [Ph3PCH20Me] + Cl ~, BuLiTHF ...... reflux, 116 .......
. ... ...

2 Ph3PCH3Br, BuLi THF reflux 116 3 Ph3PCH3I, NaH DMSO 65 116 4 Tosmic, t-BuOK, MeOH DME ta 116 + dcomp.

Tosmic, t-BuOK, MeOH DMSO ta 116 + dcomp.

6 (Et0) ZP (O) CN, LiCN THF reflux 116 7 NaH, Me3SI DMF ta decomp.

8 ICH2Cl, BuLi THF your decomp.

9 PhS (O) CH ~ CI, BuL ~ i THF -78 -20 116 + dcorne We thought that the problem may have come from the group t-butyldimethylsilyl on the molecule. So we have redone the input approval reactions 3, 5, 7, 8 and 9 on free alcohol at position C-3 and with a benzoate instead of t-butyldimethylsilyl on the molecule 116. In all cases, the same results as with t-butyldimethylsilyl have been obtained. The problem therefore did not come from this protective grouping at the position C-3 of the ketone 116 but of the molecule itself.
So we tried to certify the ketone at position C-17 through a reaction of substitution. To do this, reduce the ketone to this position by matching alcohol and transform this alcohol into a good leaving group. So we have reduced the Ö
ketone 116 with sodium borohydride and we got alcohol at 121 in one quantitative yield. We treated this alcohol with tosyl chloride for us lead, in a yield of 75%, to tosylate 122 which is a good grouping therefore.
We tried to move this tosylate with sodium cyanide in the dimethyl sulfoxide (DMSO) 66. At room temperature, we get only the starting material and by heating we get the elimination of fluorine as well only tosylate to obtain diene 123 in a yield of 77% (diagram 38). Starting from result, we deduced that the elimination reaction rate of tosylate 122 is higher than that of substitution of the latter. The corresponding olefin is therefore obtained.
Therefore, the protons in a of this olefin become more acidic. The reaction elimination of fluorine then becomes easier and that's why we get the diene 123.
Me O Me ~ H
H NaBH4, MeOH H TsCI. And N, DMAP
0 ° C, 100% CHpCl2, 75%
F ~~~ F
TBDMSO TBDMSO
116,121 Me ~ Ts Me ,, H NaCN, DMSOi H
ta at 80 ° C
HF 77%
TBDMSO TBDMSO
122 12 ~
Figure 38 Finally, all the methods we used to try to certify to the position C-17 of ketone 116 did not work. The strategy for reach aldehyde 118 (Figure 9) should therefore be modified.

4.2 Functionalization of cycle A with approval made at position C-17 The second strategy to obtain aldehyde 118 consists in functionalizing cycle A after approval of the ketone at position C-17 (Figure 24). From the nitrite 90b us could obtain aldehyde 118 (Figure 9) and synthesize butenolide present in the 10-nor-143 fluorodigitoxigenin 117 as well as analogs 119 at position C-17.
HORN
Me CN Me CHO
H
_____ ._____ Fi FHF
Me0 ~ TBDMSO
90b 118 119 i H
3; '~.
Figure 24: Plan B for the synthesis of 10-nor-14 / .3 fluorodigitoxigenin 117 and its analogous to position C-17.
4.2.1 Strategy with nitrite at position C-17 In Chapter 2, we synthesized compound 90b which contains the nitrite at position C-17 (Figure 28). So we tried to functionalize cycle A from of this compound. First, we treated compound 90b in the conditions of Birch that we optimized in Chapter 3. However, we have not added ethanol to avoid reduction of nitrite to amine. Under these conditions of reduction we we only obtained decomposition products (diagram 39). These products of decay appear to result from side reactions with nitrite to the position C-17.
Me CN
1) Li, NH3, THF, -78 ° C
2) HCI 1N, THF '~ Decomposition I ~ H) F
MeO ~ ~, / \ - ~~
90b Figure 39 So we tried to reduce nitrite 90b to aldehyde and then redo the reduction from Birch. Düsobutyalaluminium hydride (Dibal) is the most used to reduce a corresponding aldehyde nitrite6 ~. However, we know that the latter can also act like Lewis acid, complex the fluorine and cause the elimination of this latest. For prevent this side reaction from happening we did the reduction of nitrite 906 with Dibal at -78 ° C but we only obtained the product of departure. By warming up gradually the solution up to 0 ° C, we obtained a mixture separable c! ', aldehyrle124a .: ~ i 124b with the eliminated fluorine (diagram 40).
Me CN Me CHO Me CHO
Dibal, toluene r ~ 'W ~ j' ~ -78 ° C to 0 ° C ~ y H 'F 100% ~ / 1. ~ I ~.
Me0 Me0 Me0 90b 124a 124b Figure 40 Other reducers have been used in an attempt to reduce nitrite 90b to aldehyde free obtain elimination of fluorine. These reducers are presented in the table 12. In a first, we reacted the nitrite 90b with the hydride aluminum and lithium.

With this reagent, one can reduce a nitrite to aldehyde but also to amineb8.
In our case, we got neither of these two products; we have isolated only products of decomposition (entry 2). We treated nitrite 90b with L-Selectride and us only recovered the starting product (entry 3). We have made a solution lithium aluminum triethoxide hydride from hydride aluminum and lithium and three equivalents of absolute ethanol. The latter is recognized for reduce nitriles to aldehydes and not to amines6 '~. In our case, using this reducer we we only got the starting nitrite (entry 4).
Table 12: Reducers used to switch from nitrite 90b to aldehyde corresponding.
~ rv Input vww- ~~~~ Reducer '.'. "~". ~ ..... ~,. ~ ° - ~ -w ~, waves of Product reaction 1 Dibal Toluene, 0 ° C decomp.
2 LiAlH4 THF, -78 ° C decomp.
3 L-Selectride THF, ta 90b 4 LiAIH (OEt) 3 Et20, ta 90b Finally, instead of reducing the nitrite, we tried to hydrolyze it to get the ester corresponding. This ester would then be easier to reduce and there would be so less than probability of obtaining elimination of fluov- First, we treated nitrite 991b with me ~ sodium oxide in methanol and we recovered the product dep; zrt.
We then treated nitrite 90b with a saturated methanol solution with acid hydrochloric gas and we got the starting nitrite. So what either acidic or basic conditions, we failed to hydrolyze nitrite 90b for us lead to the corresponding ester.
All the methods we used to turn nitrite into aldehyde or in ester did not work. So we have to find another group than nitrite for approve at position C-17 from ketone 14.

4.2.2 Strategy with the ester at position C-17 The coupling reaction with triflic enol ether 88 to obtain the nitrile 90b works well (diagram 28, chapter 2). So we decided to keep the same type of reaction approval to change the functional group at position C-17. At place to do the coupling reaction with a nitrile, we did a reaction. of carbonylation for obtain the ester a, ~ i unsaturated 125 at position C-17 (diagram 4I).
Me OTf Me C02Me Me COzMe carbonylation hi F ~ \ Fi F ~ \ h1 Me0 ~ Me0 ~ Me0 Figure 41 The results that we obtained are presented in Table 13. All the reactions carbonylation were carried out at a temperature of 70 ° C and at a monoxide pressure 150 psi carbon with methanol. In the first two tests that we have performed (inputs 1 t; t 2), we used as base 1E ~ -carbonata of pc-ia. ~ if u, -; . ,. ..'vs two different catalysts. In both cases, we got ur_ mixture s p ~ :: ~ Maple ester a, / 3 unsaturated 125 and diene 126. The secondary elimination product of fluorine is explained by the fact that we use a strong base (K2C03) and that it is not soluble in the reaction medium. So we did two more tests of carbonylation with two different catalysts using triethylamine as the base (inputs 3 and 4). In the two cases, we obtained the desired unsaturated ester a, ~ 3 125 in returns 80% and 81% comparable and we eliminated the formation of diene 126.
The choice of the basis for the carbonylation reaction is therefore very important to avoid the reaction elimination of fluorine.

Table 13: Carbonylation tests on triflic enol ether 88.
InputCatalyst ~~~~ Nry ~~~ Y ~~~ Base ~~~~~~~ Solvent Product (rdt) ... . 1.. ...... ... ~ PdClz (PPh3) 2 ..... . KZC ~ 3 .... DMF. ...
...... 125 (40%) and 126 (25%) -...
2 Pd (PPh3) 4 K2C03 DMF I25 (63%) and 126 (16%) 3 PdCl2 (PPh3) 2 Et3N DMF 125 (80%) 4 Pd (OAc) 2, PPh3 Et3N DMF 125 (8I%) We then hydrogenated ester a, ~ unsaturated 125 with hydrogen and palladium (0) to lead us to ester 127 in the form of a single diastereoisomer (diagram 42).
We did chemical proof to determine the stereochemistry of the ester 127. A
chemical proof consists in taking an intermediary whose structure is uncertain and to the drift to a known intermediary.
So we treated ester 127 with düsobutylaluminium hydride (Dibal) and we we obtained a separable mixture of alcohols 128a and 128b in a ration of 3 : 1 (diagram 42). The majority alcohol 128a is that which contains the double bond trisubstituted, formed by the fluorine elimination reaction, and the minority alcohol 128b is the one that contains double tetrasubstituted link.
The structure of nitrite is 90a known since we have determined the structure by X-ray of its counterpart / 3 (Figure 21, Chapter 2). So we treated nitrite at 90a with Dibal and we got an inseparable mixture of 129a aldehydes and 129b.
We have reduced this mixture of aldehydes with sodium borohydride to give us a mixture of alcohols in a 3: 1 ratio (Figure 42). 1 H NMR spectra spirits obtained from ester 127 and alcohols obtained from nitrite 90a are the same.
We were therefore able to conclude by chemical evidence that the stereochemistry of the ester 127 to the position C-17 is of configuration a.

Me C02Me Me C02Me H ~, 5% Pd / C ~
F EtOH, 100% I ~ ~ Fv Me0 ~ Me0 125 127 Dibal, T oluene ~ 0 ° C, 100%
Me,:., OH Me '-.- OH
.- ..
i \ _ i H i H
Me0 ~ Me0 128a. 128b (3: 1) ~ NaBH4, MeOH
100%, 2 steps Me CCN Me ÉCHO
Dibal, Toluene Fi F 0 ° C ~ /. H
Me0 Me0 90a 129a: 129b Figure 42 In the affected products, the stereochemistry of the substituents at position C-17 is ~ 3. We have therefore attempted to reverse the configuration of ester 127 by performing a ester reduction 1-4 a, /. ~ unsaturated 125. Whether with L-Selectride or with borohydride sodium in presence of nickel, we always got only the ester at 127. We so we have continued the sequence with this isomer and we will try to epimerize it later in the sequence.
To continue the sequence, we performed Birch reduction of compound 127 with the ester has at position C-17. We got 75% ketone yield a ,, unsaturated C ~

130 with the ester at position C-17 reduced to alcohol. We have next hydrogenated olefin of this α, β-unsaturated ketone to lead to compound 131 which comprises the junction of AB cis cycles. The alcohol of this compound is then oxidized with perruthenate of tetrapropylammonium (TPAP) and with 4-methylmorpholine N-oxide (NMO) 4 ° and we obtains aldehyde 132 in a yield of 80%. When we tried to epimerize this aldehyde by making the enolate of the latter using düsopropylamide lithium (LDA), we got the starting aldehyde and decomposition products (Figure 43).
CO Me, OH
Me, 2 1) Li, EtOH, NH3 Me THF, -78 ° CHH ~, 5% Pd / C
W = = AcOEt, 67%
HF 2) HCI 1N, THF HF
Me0 ~ 75% ~
127,130 Me ~ OH Me CHO
HT AP, NMO H ~ Epimerization Starting material 4 MS, CHZCIz +
~ Fi F gp% ~~ F Decomposition O / \ O

Figure 43 By referring to similar molecules in the literature69 to find conditions of aldehyde 132, we found that the product kinetics and the product thermodynamics of this type of steroid, with the junction of CD cis cycles, is the same either the isomer a at position C-17. So, that we are trying to epimerize the aldehyde has 132 in kinetic or thermodynamic conditions, we will always get the starting product.

To have the configuration ~ i in position C-17, it is absolutely necessary to pass with a nitrile. It is the only substituent which makes it possible to obtain the center ~ 3 at this position of steroids with the junction of CD cis cycles. It is this functional grouping that we have used in the model series and which enabled us to obtain nitrile ~ i 90b (chapter 2, figure 28).
Figure 25 illustrates the attack faces of a nitrilate and an enolate in kinetic conditions.
t, e nitrilate 133 is planar and linear and curiously the protonation kinetic takes place mainly on the face a (concave face) of the molecule. Enolate 135 meanwhile, is planar but nonlinear and in this case the kinetic protonation takes place only on the face, ~ i (convex face) of the molecule to obtain the aldehyde has 136. We can therefore conclude that it is very important to have a linear anion to have a protonation kinetics on the concave side of this kind of steroid. Therefore, the nitrite is the only one pool which can provide center ~ 3 at position C-17 as demonstrated in the synthesis of digitoxigenin 1 performed by the Stork48 group.
ROH (20%) R CN
R
CSN. . /
__ ~ ~. ~ ~ JF
.., 133, ~~, ~ 134 ROH (80%) VS
ROH
Me ~ Me, CHO
O-F / C \ H ~ ~ HF

Figure 25: Protonation of nitrilate 133 and enolate 135.

So we gave up our efforts to try to epimerize the aldehyde 132 since according to our results as well as those of the literature, we would be impossible to get aldehyde 132, Q by this route.
In summary, when we homologate ketone 14 at position C-17 with a nitrile we we obtain the desired isomer, l3 at this position (chapter 2, diagram 28). Through against, from that nitrile / 3, we cannot functionalize cycle A. On the other hand, when homologous this same ketone 14 with an ester we can functionalize the cycle A
but we cannot get the ~ isomer at position C-17. Must therefore go back to first synthetic plan that we proposed at the beginning of this chapter (figure 23). This plan consists in functionalizing cycle A before doing the homologation at the position C-17.
4.3 Strategy with vinyl iodide and functionalized cycle A
We have tried several types of approval reactions on ketone 116 which contains cycle A functionalized. In any case, we never got the desired product (table 11). We also tried to synthesize the triflic enol ether of ketone 116 to do a coupling reaction but we had no success (table 10). During the study reshapes (chapter 2), the only method that has yenn ~:
sy ~; tlétiser nitrile à la position C- ~ 1 '7, is a coupling reaction with palladium.
Fn considering these results, we deduced that the coupling reaction is the method suitable for approving the ketone at position C-17 in nitrite. However, it must find another group than the triflic enol ether to perform this reaction of coupling. In the literature, there are several examples of this type of reaction with iodides vinyl '°. So we synthesized the vinyl iodide of the ketone 114 for then carry out the approval reaction at position C-17.
To synthesize vinyl iodide7 ~, we reacted ketone 114, which contains the cycle A functionalized, with hydrazine hydrate and with triethylamine during 5 days.

The corresponding hydrazone is thus obtained which is treated with iodine and triethylamine to lead us to vinyl iodide 137 in a 91% yield for two step.
It is then necessary to carry out the homologation reaction on this compound. For this do we have treated vinyl iodide 137 with tetrakis (triphenylphosphine) palladium (0), the cyanide of lithium and ether crowns 12-C-4. We thus obtain the nitrile a, ~ unsaturated 138 desired with a yield of 40%. We could not get this pure compound because of the presence of secondary products which co-elute during chromatography-flash. To try to optimize the reaction we changed the palladium catalyst by the diacetate .de palladium (II) but we only got the starting product. In changing the cyanide of lithium for potassium cyanide we got products from decomposition (diagram 44).
OI
Me ~) NH2NH2.H20, Et3N Me H EtOH, reflux H
2) 12, Et3N, THF
HF g1%, 2 stages Fi F
HO HO
114,137 Pd (PPh3) 4, KC ~ ~ ~ Pd (PPh3) 4, LiCN

18-C-6, benzr Pd (OAc) 2, PPh3 I 12-C-4, t7enz LiCN. Et 3 N; DMF ~ ~~ 40%
Me CN
Decomposition Starting material H
HO
138 (unclean) Figure 44 To improve the coupling reaction, we protected the alcohol at the position C-3 of vinyl iodide 137 in t-butyldimethylsilyl form (compound 139), yield quantitative) and we redid the coupling reaction. We have acquired nitrite a "(3-unsaturated 140 pure in a yield of 45%. We then hydrogenated the olefin of this compound and we got 53% of the nitrite ~ 141b and 47% of the nitrite at 141a (diagram 45).
Me I Me TBDMSOTf,, Et ~ N_ H ~ _Pd (PPh3) 4, LiCN
Fi F CH2CI2, 100% HF 12-C-4, benz HO TBDMSO 45%
13ï 139 Me CN Me CN Me ÇN
H2, 55% PdIC
AcOEt p ~ / ~ FHF h1 F
TBDMSO TBDMSO TBDMSO
140 141b (53%) 141a (47%) Figure 45 To prove the stereochemistry of nitrite ~ 3 141b, we hydrolyzed ether silylated from this last with p-toluenesulfonic acid in methanol to get alcohol 142b corresponding in quantitative yield. We have tried several systems solvents to: install this alcohol but we have not obtained crystals. We have so treated alcohol 142b with p-nitrobenzoyl chloride for us lead to p-nitrobenzoate 143 (scheme 46). These derivatives are known to crystallize easily. We we therefore obtained crystals of p-nitrobenzoate 143 by diffusion of pentane in ethyl acetate.

Me CN Me CN
_ APTS. MeOH_ HO ~ NCRH9COCI, pyr HF 100% HF ~ DMAP, CH2CI2 TBDMSO HO
141b 142b Figure 46 The X-ray structure of this compound is shown in Figure 26. We can note that the nitrile at position C-17 is good ~, that the alcohol at position C-3 is ~ 3 and that the junction of the cycles AB is well cis. We therefore confirm beyond any doubt the stereochemistry of all our centers.
Figure 26: X-ray structure of nitrile /.3143.

t02 4.4 Synthesis of butenolide at position C-17 4.4.1 Synthesis of butënolide by Stork In the literature, butenolide at position C-17 of digitoxigenin 1a been synthesized by Stork4 $ from nitrite 144. The Stork sequence is presented in the figure 47. In a first, alkylation with lithium (benzyloxy) methyl gives the compound 145 in a yield of 89%. The hydrogenolysis of the benzyl group is then done to obtain hydroxyketone 146. The latter is treated with the ketene of triphenylphosphoranylidene to form butenolide at position C-17. Stork so gets butenolide with a yield of 74%. Finally, the hydrolysis of ether silylated in the middle acid of this compound is carried out to lead to digitoxigenin 1. The sequence for switching from nitrite 144 to butenolide therefore takes three steps and the yields are very good.
Me CN
Me ~ BuLi, BnOCH2SnE I ~ 10% Pd / C
f-1 'OH THF, -78 ° C, 89% ~ tOH, 96 °. ~ 0 TBDMSO

H
1) Ph3P = C = C = O, Et3N
benz., 74%
2) APTS, MBOH, 97%
TB

Figure 47 4.4.2 Synthesis of butenolide from nitrile There are two differences between the nitrite 141b that we synthesized (Figure 45) and nitrite 144 that Stork synthesized. In our case, we have fluorine at the C- position 14 instead of hydroxyl group and we have hydrogen at position C-10 instead of methyl. AT
a priori, these differences should not cause us any problem for the synthesis of butenolide at position C-17.
First, to test the conditions developed by Stork48, we used the mixture of nitriles 147 (2: 1, ~ 3: a) 4 ~. So we treated this mix in the same alkylation conditions that Stork and we only got the product of departure.
Even at higher temperatures, there is no reaction that occurs (diagram 48).
We then tested the alkylation reaction conditions on the nitrite at 141a and we we obtained the starting nitrite in the presence of decomposition products.
We have also treated nitrile, Ci 141b under the same conditions and we obtained a separable mixture nitrite at 141a and nitrite / d 141b and decomposition products (diagram 48). We We therefore concluded that the (benzyloxy) methyl of lithium came to make the nitrilate of the three nitriles that we used in li ~, zi to do the ~acti ~~ n alkylation as in the sequence of Stork4R, so you have to find another way to synthesize Ie butenolide.

Me CN
BuLi, BnOCH2SnBu3 _ Starting material \ H Hv THF, -78 ° C at ta me0 147 2: 1 (/ 3. A) Me, CN
H BuLi, BnOCH2SnBu3_ Starting material 'H ~ t' F THF, -78 ° C Decomposition 'TBDMSO
141a Me CN Me, CN
H BuLi, BnOCH2SnBu3_ H Starting material + +
hl F THF, -78 ° CHF Decomposition TBDMSO TBDMSO
141b 141a Figure 48 4.4.3 Synthesis of butenolide from aldehyde The alkylation of lithium (benzyloxy) methyl on the aldehyde at position C-17 instead of nitrite would be a good way to work around the problem. However, we know that the reduction of nitrite 90b with düsobutylaluminium hydride, gives us elimination of fluorine (diagram 49).

Me CN Me CHO
Dibal, toluene _ - -78 ° C to 0 ° C ~ ' HF 100%
Me0 Me0 90b 124 Figure 49 So we tried to find other conditions for reducing a nitrile in aldehyde in using as a model the mixture of nitriles 1474 (figure 27).
Me CN Me CHO
_ Reduction_ _ H Fi ~ \ Fi H
Me0 Me0 147 2: 1 (~: a) 148 Figure 27: Reduction of the nitrile mixture 147.
l., es the different reaction conditions as well as the results, obtained are presented in the table 14. First, we tried to make the nitriliums of the mix of nitriles 147 and reduce them with a hydride source ~ 3. So we have processed the mixture of nitriles 147 with triethyloxonium tretrafluoroboronate and we have retrieved on starting material and decomposition products (entry 1). We have then tried two different reduction conditions with Raney'4 nickel. In one case, We have obtained the starting product (entry 2) and in the other case, decomposition (entry 3). We also tried to reduce the mixture of nitriles 147 with the hydride aluminum and lithium at different temperatures. Ä -78 ° C, the starting material in summer recovered (entry 4) and at 0 ° C, decomposition products were recovered (entry 5). With aluminum triethoxide hydride and lithium, we only got the product of departure (entry 6).

Efforts to find a reducer other than Dibal to reduce the nitrile mixture 147 were unsuccessful. So we reduced this mixture of nitriles to aldehydes with the Dibal to test the alkylation conditions of (benzyloxy) methyl lithium. We have obtained the mixture of aldehydes 148 in a yield of 60% (entry 7).
Table 14: Conditions for reducing the nitrile mixture 147.
Between Product Solvent Reagents (rdt) ____ 1_ ~ ...___. ~~ __l ~ Et3O + BF4 ~, ~ CH2C12.-..._ .._ ~ _._ 47 + -dcomp.
_ 2) Et3N _._.

2 Ni / Raney. 75C HCOZH dcomp.

3 Ni / Raney, NaHzP02.2H20 Pyr: H20: HOAc (2: 1: 1) 147 4 LiAIH (OEt) 3, 0C ta THF 147 LiAlH4, -78C THF 147 6 LiAlH4, -0C THF dcomp.

7 Dibal, -78C Tolune 148 (60%) We then tested the alkylation reaction on the mixture of aldehydes 148. For this do we used the same conditions as those used with the mixture of nitriles.
We thus obtained a mixture of alcohols which we oxidized to lead to a mixture of ketones 149 in a yield of 63 ° rb for the two stages (sc-héma SO). DONV.
passing through aldehyde instead of nitrite, the alkylation reaction of (benzyloxy) methyl of lithium works well on the model compound.
Me CHO 1) But.i, BnOCH2SnBu3 THF, -78 ° C
2) TPAP, NMO, 4 R MS
Me0 I / HH CH2CI2 148 2; 1 (~: a) 63%, 2 steps 149 2: 1 (aa) Figure 50 We must now find a method to reduce the nitrite ~ i 141b without eliminate fluorine and do the alkylation step on the corresponding aldehyde. With the compound model 147, the Dibal is the only reagent that has made it possible to obtain the aldehyde from nitrite (Table 14). We we therefore optimized the reduction conditions with Dibat on nitrite a 141a because this the latter is the unwanted isomer. To start, we treated nitrite a 141a with the Dibal at -78 ° C and we obtained only traces of the aldehyde at 150a and the product of departure. We therefore warmed the reaction mixture to -60 ° C and after an hour the reaction is complete. After purification on silica gel, we obtain Aldehyde at 150a in a quantitative yield which always contains fluorine at position C-14. We have found that this aldehyde is unstable even at low temperatures and under inert atmosphere. he we now have to carry out the alkylation reaction on this aldehyde.
To do this, we reacted aldehyde 150a with lithium (benzyloxy) methyte and we do not have obtained only from decomposition products (diagram 51).
Me, CN
Me, CHO
1) Dibal, toluene, -60 ° C
H
~ \ ~ r ~ F ~ 1 _, 2) silica / ~ ~ I '~ H JI 100% HF
TBDMSO TBDMSO
141a 150a, BULi, BnOCH ~ SnBu3 THF, -78 ° C
Decomposition Diagram 51 Despite this bleak result, we reduced nitrite, l ~ 141 b with the Dibal at -60 ° C and after purification, we obtained a mixture of approximately 1:
1 of aldehydes a and ~ 3.
The aldehyde / d 150b therefore epimerizes on silica to aldehyde at 150a, the product thenmodynamique69. The purification step on silica is very important since this is where when you completely hydrolyze 1 "imine which is formed when reduction. For minimize the dehydration of the aldehyde ~ 31501y, we must do a very small silica column and very fast elution. We thus obtain an inseparable mixture of aldehyde ~ 150b and aldehyde at 150a in a ratio of 9: 1 respectively. We have then do the alkylation reaction on this mixture of aldehydes and we got that products of decomposition as in the case of aldehyde a (diagram 52).
Me CN Me CHO
H ~ 1) Dibal, toluene, -60 ° CH
2) silica hi F 100% HF
TBDMSO TBDMSO
141 b 150b: 150a 9: 1 (%: a) But.i, BnOCH2SnBu3 THF, -78 ° C
Decomposition Figure 52 We can see that despite the great similarity between nitrite 144 synthesized by Stork ~ z (diagram 47) and nitrite 141b which:, we have synthesized (diagram 4r), we ra '~~~, ~ ons could not get butenolide at position C-17 by this method. The alkylation reactions with the lithium (benzyloxy) methyl which were made on the nitrite ~ i 141b and nitrite a 141a or on the aldehyde ~ i 150b and the aldehyde a 150a gave us that products of decomposition. So we put aside the synthesis of butenolide at the position C-17 to focus on the synthesis of analogs to this position.
4.5 Synthesis of analogs at position C-17 We saw in the introduction that the analogues at position C-17 of the digitoxigenin 1 have biological activities comparable to the latter (introduction, figure 3) 3.

In most cases, biological activities are always higher when the center in C-17 is /.i. The analogs at position C-17 are a, ~ unsaturated esters with different chain lengths. It would therefore be interesting to synthesize the same analogues but with a fluorine at position C-14 and compare the biological activities obtained.
from the aldehyde at 150a and the aldehyde, l3 150b that we have synthesized we lungs get analogs at position C-17 of digitoxigenin 1 with fluorine position C-14. As a starting point for comparing biological activities, We have synthesized a, / 3-unsaturated methyl esters from these aldehydes.
So we treated the aldehyde at 150a and the aldehyde, l3150b separately with the anion of trimethyl phosphonoacetate to obtain the esters a ,, (~ unsaturated 151a and 151b respectively with the geometry of the trcrns double bond exclusively, this which was determined by 1 H NMR (diagram 53).
CHO Me H NaH, ~ MeO) ~ P (O) CH ~ C02M
THF
hi F
TBDMSO
150t ~ C. - 9: 1 (/3.a) non-separable 151b C-17 = r: ç (~.; Xi r ~ on-separable 150a C-17 = a 151a C-17 = a Me APTS, MeOH, 152b C-17 = <9: 1 (p..a) non-separable 152a C-17 = a Diagram S3 These esters are then treated a, ~ unsaturated separately with the acid p-toluèsulfonique for hydrolyze the silyl ethers and we get the alcohols 152a and 152b respectively (diagram 53). At this stage, with the ester a, / j unsaturated, ~ 3152b, we observe a little epimerization of the center in C-17 since it is the a isomer which is the most stable thermodynamically To avoid the epimerization of the center in C-1 ~, we have reversed the steps doing hydrolysis of silylated ether before reduction of nitrite. So we have treated nitriles 141a and 141b with p-toluèsulfonic acid in methanol to obtain the alcohols 142a and 142b in a quantitative return for each. We then reduced the nitrite of these compounds with düsobutylaluminium hydride and we got the aldehyde at 153a pure and the aldehyde ~ 153b which contains 10% of the isomer a (due to epimerization on silica).
We made the Wittig reaction with the anion of phosphonoacetate of trimethyl on these aldehydes and we got esters a, ~ unsaturated 152a and 152b in a yield of 60% for the last two stages (diagram 54).
Me CN Me CN
H ~ APTS, MeOH, H ~ 1) Dibal, toluene, -60 ° C
100% ~ 2) silica Fi ~ F v ~ 'H ~ F
TBDMSO ~ y '' HO
141 b C-17 =, ~ "9 42b C-17 = ~ i 141aC-17 = a 142aC-17 = a Me ÇHO
H NaH, (MeO ~ P (O) CH ~ COZMe THF, 60%, 2 steps HF
HO
153b C-17 = 9: 1 (A: a) non-separable 152b C-17 = 9: 1 (/ ~: a) non-separable 153a C-17 = a 152a C-17 = a Figure 54 In summary, we have combined the model studies we have carried out in the chapter 2 and in chapter 3 with some modifications. The best combination was to make the functionalization of cycle A followed by approval at the position C _'.- 17 by the way with vinyl iodide instead of triflic enol ether. To get the center ~ 3 at the position C-17, it is absolutely necessary to go through a coupling to form a nitrite and so we we obtain nitrite ~ 3141b as well as nitrite at 141a (approximately 1: 1). AT
from these nitriles, we were unable to obtain butenolide at position C-17 of 10-nor-14,13 fluorodigitoxigenin 117. However, we were able to obtain the ester a, /. ~ insab. ~ ré at I52a and ester a, ~ unsaturated ~ i 152b which are analogs to position C-17 of the 10-hor-14 /. ~
fluorodigitoxigenin 117. These analogues were obtained in 36 steps in a yield 87% per step and with an overall yield of 0.7%
The biological activities were measured on the ester a ,, (~ unsaturated at 152a pure and on 9: 1 mixture of esters a "L ~ unsaturated 152b containing the isomer, Q so majority. The results of these tests will be presented in the next chapter.
Diagrams SS to 58 summarize the sequence for synthesizing analogs 152a and 152b of the 10-rzor-14 ~ -fluorodigitoxigenin 117.
y 1! Bones, lüeGH, -78 ~ <~
1) LiAIH4, THF, -78 ° C ~ ~. ~ 2) NaBH4, ta ~.
'2) APTS, benz, reflux y, ~ 3) NaOH, H202, THF
Me0 ~ Me0 21 100% 27 86%
1) Mn02, pent: CH2Cl2 85%
OH 2) TBDMSCI, Im. ~ ~ H
Me0 OH THF, 100% Me0 ~ OTBDMS

Figure 55 O 1) (Et0) 2P (O) CH2C02Et OH
H NaH, THF, 97%
OTBDMS 2) Dibal, CH2CI2, 0 ° C
Me0 97% Me0 ~ I OTBDMS

1) TBDPSCI, Im ~~ OTBDPS
THF. 93% ~ n-BuLi, THF, -78 ° C _ 2) PPTS, EtOH, 98% ~ (Et0) P (0) CH (F) CO2Et 28 3) Swern, 100% Me0 H 93 / o ~ OTBDPS I ~ OTBDPS
/ IF 1) LiAIH4, THF, 0 ° C, 100% _ 2) HCA, PPhg, THF, 99% ~ IF
Me0 C02And Me0 CI

Nal, NaH, n-BuLi _ J 1) TBAF, THF ~ 77%
MeC (O) CH2C02Me, "~ °, F ~) PivCl, 2.6-lut THF, 55% 1vIe0 ~~ \ ~ - '' J ~ 'v._ ~ - ~ ;;. ~, ~ L,. Rome CH2CI2:
97 ° ~ °
° ' EO
~ OPiv pd (PPh3) a, Ph2P (CH2) 3PPh ~ IF ~ '.
BSA, THF, reflux, 2 * 10-3M
F
Me0 ° ~ OMe add, slow (2 p.m.), 88% Me0 OTBDPS
Figure 56 EOEO
F (PhSeO) 20, Et3N \ F °
And ~ N, toluene, 165 C
CHpCl2, 91%
4 h, sealed tube Me0 ~ Me0 ~ 78%

EOH 1) Bu2Sn (O) benz: DMF 13: 1) 1) H2, 5% Pd / BaS04 _ reflux _ -EtOJ-I absy 100% 2) TsCI, ta, 87%
HF 2) LiBH4: MeOH (1: 1) MeOr ~ THF, 94%

10: 1 a. ~
) H
Me ~ OH
NaBH4, DMSO 1) Li, NH3, EtOH, THF
80 ° C, 85% ~ I ~ _ -78 ° C to -30 ° C
HF 2) HCI 1N., THF
Me0 ~ 77%

9: 1 (a:, (~
OH O
Me _, "~ e ~;
1) Hz, 5% Pd / G
N r ~ '' __AcOEt, 60% H L-3 electrade, 2) TPAP, NMO = '"~ THF, 100%
HF 4 to MS, CH2CI2 ~~~ F
99% O

Where i Me 1) NHZNH2.H20, Et3N Me H EtOH, reflux H \
2) IZ, Et3N, THF
HF 91%, 2 steps HF
HO HO

Figure 57 Me I Me CN
1) TBDMSOTf,, Et3N
H ~ CH2Cl2, 100%; Hue, 5% Pd / Ca HF 2) Pd (PPh3) 4, LiCN = AcOEt 12-C-4, benz, 45% HF
HO TBDMSO
133,136 Me CN Me CN
APTS. MeOH ~ H 1) Dibai, toluene, -60 ° C
H 100%
2) silica HF hi F ~
TBDMSO HO
137b 53% C-17 = R 138b C-17 = / 3 137a 47% C-17 = a 138a C-17 = a CHO Me H
NaH, (Me0) ~ P (O) CHZCO ~ Me HO ~ HF THF, 60%, 2 steps 147b C-17 = 9: 1 (p: a) non-separable 147a C-17 = a 146b ~ '-17 = 9: 1 (~ i'.a) non-separable 146a ~. "~ -17 = a Figure 58 LILY
Chapter 5 Synthesis of l4 ~ fluorodigitoxigenin and biological activities This chapter describes how we synthesized the 14 ~
fluorodigitoxigenin 7 to starting from digitoxin 2. He also describes the synthesis of a fluorinated analogue to position i: -17 of the latter via the degradation of butenolide at this position. We have also compiled all the biological activities of the compounds we have obtained.
5.1 14 / ~ Fluorodigitoxigenin The idea of synthesizing Ia 14.13 fluorodigitoxigenin 7 from the product natural came to us side reactions that we observed during our sequence for synthesize the flurosteroids 152a and 152b via the transannular Diels-Alder reaction. Through example, with boron tribromide, we got the exchange of fluorine at position C-14 for a bromine, with Dibal and Tebbe's reagent, we got the products elimination fluorine and under the Birch reduction conditions we got hydrogen at the position C-14 in place of fluorine. From these results, we have assumed that the same kind of reactions might preclude with the hydroxyl group at the position C ~ '-1: ~ cic-' l ~~
digitoxigenin 1.
So we did some research on this type of reaction and we found an article in the literature which describes the fluorination of 3-O-acetyldigitoxigenin 154 at position C-14 performed by Kortmann's team ~ 4. To do this, they process this last with potassium fluoride and sulfur tetrafloride and they get the cUmposed 155 with the fluorine at position C-14a and olefin 156 (Figure 59).

. SFa_ ~ 2 ~ zC
154,155 (17%) 156 (16%) Figure 59 The stereochemistry of fluorine at position C-14 was determined by 19F NMR in calculating the coupling constants between fluorine and its protons.
Fluoride absorption is a split triplet with a coupling constant of 26 Hz for the triplet and a 8 Hz coupling constant for the doublet. Fluorine at position C-14 is so coupled with two axial protons (at 180 °) which gives a triplet with a great constant of coupling and with an equatorial proton which gives a small constant of coupling. Through therefore, the fluorine at position C-14 must be of stereochemistry a so that the NMR signal l9F agrees with the structure of the molecule.
To be certain of the stereochemistry of fluorine deduced by 19F NMR, we wanted to redo the fluorination reaction in the same conditions then f ~ irP an X-ray of the product got. Unable to readily obtain sulfur tetrafluoride, we we used different conditions to do the fluorination reaction either with the trifluoride (diethylamino) sulfide (DAST) ~ 5.
To start the sequence, we hydrolyzed the trisaccharide from digitoxin 2 with sulfuric acid in methanol to give us the corresponding alcohol.
We have then treated this alcohol with acetic anhydride in pyridine to get Ia 3-O-acetyldigitoxigenin 154 in a yield of 89% for the two stages ~ 6. In dealing with this last with a mixture of DAST ~ S, HF.pyr and pyridine in the dichloromethane we got compound 157 with fluorine at position C-14 in a yield of 18% (diagram 60) as well as the alcohol elimination product 156. It is necessary point out that the fluorinated compound 157 is obtained in the form of a single isomer according to NMR analysis 1H. We then hydrolyzes the acetate of this fluorinated compound 157 with a few drops acid concentrated hydrochloric acid in methanol and the 14-fluorodigitoxigenin 7.
H2S04, Me0 EST, HF.pyr AC20, pyr ~ r, CHzGI
9%, 2 stage: 18%

H
HERE conc, MeOH
82%

Schéana 60 We crystallized this product by diffusion of pentane in acetate ethyl. The X-ray structure shown in Figure 28 confirms beyond doubt that fluoride at position 14 is of stereochemistry ~. We have ~: we therefore synthesized the 14 / ~ fluorodigitoxigenin 7 in four steps from digitoxin 2.

Figure 28: X-ray structure of l4 ~ fluorodigitoxigenin 7.
To obtain fluorine of stereochemistry ~, it is necessary to pass by the carbocation 158 at position C-14. Looking at a molecular model, we can see that this carbocation is not totally planar. It has a slight convex (top side) and concave face (face of below). The attack of fluoride is therefore done on the face of the top of the molecule 158 (face convex), which is the least crowded side, to lead us to the compound fluorinated 157 (figure 29).
F-Me OO Me, .. ~ O ~ GO
Me _- Denies.
, _ - +
l Ac0 ~ 7 F-Figure 29: Fluoride attack face.
We did 19F NMR of fluorinated compound 157 (C-3 alcohol protected under form acetate) that we synthesized and we compared it with the spectrum fluorine compound 155 obtained by the group of Kortmann ~ 4. In our case, we get a peak for fluorine which corresponds to a multiplet and in their case with fluorine a they get a triplet split. Therefore, this means that we have synthesized the isomer, Q
and that they have well the isomer a. So, depending on the reaction conditions used, we can get the fluorine at position C-14 of stereochemistry a or ~. So we are the first to have synthesized 143 fluorodigitoxigenin 7 from the natural compound and this using the DAST reagent.
We measured the biological activity of l4 ~ Fluorodigitoxigenin 7.
So, we will ability to compare result obtained with biological activity of compound natural be it digitoxigenin 1. Biological results will be presented later in this chapter.
5.2 14, Q-Fluorodigitoxin We also carried out the synthesis of 14 / .3 fluorodigitoxin 161 from digitoxin 2. First, we protected the alcohols from the trisaccharide under form of acetates and we got compound 159 in a yield quantitative ~ 6. We have then treated this compound under the fluorination conditions used previously and we we obtained the fluorinated compound 160 in a yield of 32% (diagram 61).
The reaction conditions for trying to deprotect alcohols from sugar are described in the Table 15. With dan sodium hydroxide, methanol, we obtained cap ia decomposition (entry 1). Using an ammonia solution in the methanol, after cruis days there was still an alcohol that was not unprotected and in continuing the reaction more for a long time, we got decomposition (entry 2). The conditions of reaction with potassium cyanide (entry 3) and lithium borohydride (entry 4) we also have given only decomposition products.

, ~ 0, reflux) AST, HF.pyr _ 100% ~ yr, C1; ~ 2C ~ 2 32%
CH3p CH30 p 159 HO Ac0 O
_ OH 3 OAc CH30 ~ v CH3O ' Ac0 OOAc H OOH

Figure 61 '' AbTeau 15 : Conditions of protection of the compos fluox 16f1.

wEnter. ~ _. "... _..._. ~ .. ~., ... Conditions .. .T ._. ~ ~ ..
M. . ... of reaction. ~ '. .. ..._. ... Product: ~
. _. _ ~ ,.

_l. . _ .. _ _._.._ _.-NaOH .1.M, MeOH ... ~ Comp, ._ _._ ..

2 NH3 / MeOH dcomp.

3 KCN, EtOH, decomp. Reflux.

4 LiBH4: MeOH (1: 1), THF dcomp.

To resolve the problem of protective groups, we have protected alcohols Digitoxin 2 sugar in the form of trichloroethyl carbonates (instead of overheads) and we we obtained 79% of the compound 162'6. We redid the fluorination reaction So last and we obtained fluorinated compound 163 in a yield of 29%.
We have then treated the latter with zinc in acetic acid and we have got 63% of the 14/3-fluorodigitoxin 161 (scheme 62). We measured biological activity of the last compound and the result will be presented later in this chapter.
, CCH ~ OC (O) CI DAST, HF.pyr pyr, CH2CIZ
pYr, 79%
29%

2 ~ O ~ - 162 H OOH GP OOGP / GP = CI3CCH20C (O) Zn. HOA _ ~
63%

163 ~ ~ 161 GP- ~ OOGP GP = CI3CCH20C (O) H OOH
3 ~ 3 Figure 62 5.3 Analog at position C-17 with fluorine at position C-14 We also synthesized an analog at position C-17 from the 14 ~
Fluorogigitoxigenin 7 obtained previously (diagram 60). To do this, we we degraded the butenolide at position C-17 in aldehyde and we did a reaction of Wittig on this latest.
To start the degradation of butenolide ~~, we treated the l4 ~ fluorogigitoxigenin 7 with ruthenium oxide and sodium periodate. We then did react the intermediate formed with sodium borohydride and we got the mix of corresponding diols. Finally, we treat this mixture of diols with the sodium periodate to make an oxidative cleavage and we get aldehyde 164. These three reactions are of a trait that is to say "one pot". We then did a Wittig reaction on this aldehyde crude with the trimethyl phosphonoacetate anion and we got the compound 8 in a yield of 29% for the four stages (diagram 63). We have measured the activity biological of this compound and the result will be presented in the section next.
CHO Me 1) NalOa, Ru04, acetone z) NaBH4, MeOH Me 3) Nai04 HF
H
HO
7,164 NaH, (Me0) 2P (O) CH2C02Me "
THF, 29%, 4 steps Figure 63 5.4 Biological activities As we saw in the introduction, the main biological activity2 glycosides their ability to increase the strength of contractions myocardial (effect positive inotropic). This positive inotropic effect results from the inhibition of the enzyme Na +, K + -ATPase membrane responsible for maintaining high concentration in Na + and in Kt in the intracellular fluid. This inhibition has the effect of regulating problems cardiac arrhythmia and hypertension.

The goal of my project was therefore to synthesize 14 /. ~ Fluorosteroids and measure their biological activity to determine their inhibitory power on the enzyme Na + membrane, K + -ATPase. Depending on the biological activities obtained, we will be able to determine if the hydroxyl group at position C-14 is a bridge donor or acceptor H because the fluorine can only be an H bridge acceptor So we measured the biological activity of the fluorosteroids that we have synthesized and we compared these values with that of digitoxigenin 1 and the digitoxin 2. The measures for fluorosteroids and the two natural compounds were performed on the same receiver and under the same conditions. We measured the interaction direct between ligands (fluorosteroids and natural compounds) and the enzyme (receptor). The results are shown as ICSO in Figure 64. This measure of activity represents the necessary concentration of the drug to inhibit 50% of the membrane enzyme Na +, K + -ATPase. The smaller its value, the better the inhibition.
Looking at the biological activities of fluorosteroids in general, we can find that they are all slightly lower than those of hydroxysteroid. The 14 ~
fluorodigitoxin 161 has an ICSO of 3.8 pM while digitoxin 2 has an 1.6 pM ICSO.
P- ~~ ur which is l4 ~ fluorodigitox.igenin 7, it has v.mP ICsn of -7.1 p, M and the Igitoxigenin 1 has an ICSO of 0.35 µM. We can therefore conclude that the fluorine is not a perfect bioisostere of the hydroxyl group at position C-14 since the activities of fluorinated analogs 7 and 161 are different from those of natural compounds.
On the other hand, it is important to note that the biological activities of fluorosteroids 7 and 161 are of the same order of magnitude as those obtained with natural compounds 1 and 2. So, synthesized fluorosteroids may prove to be substitutes effective natural hydroxysteroids insofar as their toxicity is lower.
More studies surges should be carried out in order to assess this toxicity.
So there are two different conclusions that we can draw from the decreased activity biological obtained for 14 ~ i fluorodigitoxin 161 and for 14 ~ -fluorodigitoxigenin 7.

First, we could conclude that the hydroxyl group at the position C-14 is an H bridge acceptor since it retains some activity. Through against, with the fluorine at position C-14, the affinity with the receptor would be less. Through therefore, the H bridge formed at this steroids position would be weaker which would probably due to incorrect alignment of fluorine in the receptor.
O

2 X = OH ICSO = 1.6 E ~ M 1 X = OH ICSO = 0.35 ~ M
H ~ OOH 161 X = F IC5o = 3.8 ~ M 7 X = F ICSO = 7.1 ~ M

H
4 X - OH iCSO = 0.35 ~ M 152b C-17.3 ICSO '= not c ~ "activated 8 X = F lCSp = no activity 152a C-17a IC5o = 12 E ~ M
Figure 64 Another possible conclusion to explain this drop in activity organic would be that fluorosteroids bind to the receptor differently than hydroxysteroids and what despite their great similarity. An example of this kind of phenomenon ~ 8 was got with folic acid 165 and its analog, methotrexate 166 (Figure 30).
Despite the great similarity of these two compounds, it has been demonstrated by crystallography that both molecules don't bind to the receptor in the same way. However, despite this difference, they both have similar biological activities for the same receiver.
~ N NYNH2 H02C - ~ I ~ IN
NC ~ ~ NN
~ H p H NH2 folic acid 165 HOZC ~ N '~! N
H ~ \ ~ N NH

methotrexate 166 Figure 30: Structure of folic acid 165 and methotrex.ate 166.
l, es t ~ é.sa ~ ltats the most surprising are those obtained with:
fiac ~ rostér ~ ai: containing the esters a, f .. ~~ -unsaturated at position C-17. We thought we'd get say biological activities higher for ester a, ~ unsaturated ~ 3152b and 8 than for ester a, ~ i unsaturated at 152a but It was not the case. The fluorinated analog 152a, which includes the ester a, ~ in.saturé a at position C-17, gives a biological activity of 12 ~, M and the fluorinated analogs 152b and 8, which have esters a, / ~ unsaturated ~ 3, have no biological activity (diagram 64).
In the literature, the biological activity of hydroxysteroid 4 (diagram 64), which includes the ester has, ~ unsaturated, the ~ is reported and it is 0.35 ~ M3. So in replacing the hydroxyl group at position C-14 with fluorine, we completely lose the activity of the molecule (compounds 8 and 152b). On the other hand, when the ester has ,, 13 unsaturated is a, we measure a biological activity (compound 152a, 12 pM). From these results, we can conclude that the fluorosteroids which contain the esters a, ~ unsaturated at the position C-17 ~ i bind to receptor in a different way than fluorosteroids which have a butenolide position C-I7 despite their great similarity. Regarding the fluorinated compound 152a with the center has at position C-17, there are two possible conclusions. In the first case, the compound 152a can bind to the receptor in the same way as hydroxysteroids or well, it can bind in a different way (as for compounds 152b and 8 with the center ~ 3 at position C-17) while retaining a biological activity of 12 p, M.
Finally, to find out how the fluorosteroids 7, 152a and 161 bind with the receiver, further studies are needed.
In conclusion, we have synthesized I4 ~ 3 fluorodigitoxigenin 7 and 14 ~ -fluorodigitoxin 161 from digitoxin 2 using the reagent DAST. By this method, we also obtained the fluorinated analog 8 comprising an ester a ,, Q-unsaturated to the position C-17.
We also measured the biological activities of our fluorinated compounds and we know it compared with. to those obtained with hydroxysteroids. The 14/3 fluorodigitoxigenin 7, 143 Fluorodigitoxin 161 and fluorine analog 152a (with ester a, /. ~ unsaturated a at the position C-17) gave us interesting biological activities (7.1 pM, 3.8 pM and .i2 p, M respectively) while the fluorinated analogs 152b and 8 (with the ester a, ~ unsaturated ~ 3 to position C-17) gave no activity to our surprise.
From these results, it will be interesting to make other analogues fluorinated digitoxigenin 1 and digitoxin 2 with different chains at the position C-17. Moreover, we can extend this fluoridation method to other hydroxysteroids natural and thus obtain other types of interesting fluorosteroids to be tested.

General conclusion First, the synthesis of ~ 3 keto ester 18 was carried out with excellent yields. We then macrocycled this, Q keto ester to give us the thirteen-member macrocycle 15 with the geometry of TCC olefins. We have performed the Transannular Diels-Alder reaction (DATA) on this macrocycle for us lead to steroid tetracycle 16 with the geometry at the junctions of the cycles formed TAC
desired.
So, from DATA's reaction, four new asymmetric centers are created simultaneously with excellent control which shows us the power of this reaction.
Then we did two model studies to functionalize the tetracycle 16. In the first study, we approved the ketone at position C-17 of our steroid and in the second study we functionalized cycle A with the proton at the position C-10 at instead of methyl which is found in 14, Q-fluorodigitoxigenin 7.
We have completed these two model studies to lead us to two analogues of the 14.13 fluorodigitoxigenin 7 or the esters a, ~ unsaturated at position C-17 152a and 152b. We we measured the biological activities of these two compounds.
Finally, we obtained 143 fluorodigitoxigenin 7 from the digitoxin 2 in using DAST reagent. We have also synthesized the 14, CBfluorodigitoxin 161 as well that the fluorinated analog 8 (ester a, ~ i unsaturated at position C-17j from this method. We we measured the biological activities of these three compounds.
The 14 ~ fluorosteroids 7, 152a and 161 gave us activities interesting biological while there is no activity that was measured for the 143 fluorosteroids 8 and 152b.
To determine the exact mechanism of action of these fluorinated compounds, other studies should be done along with other fluorosteroids.

Experimental part Overview All solvents were dried and distilled before use. Table 16 summarizes the different conditions for the preparation of reagents and anhydrous solvents.
Table 16: Conditions for the preparation of anhydrous reagents and solvents Solvent / Reagent ~~~~~~ tt '~~ a ~ p ~ - ~ w ~' ~ Desiccant . . ... _. . .Dichloromethane. ... ...... Calcium hydride ...... .....
... ....
Düsopropylamine Calcium hydride Sodium ether, benzophenone Methanol Magnesium, iodine Tetrahydrofuran Sodium, benzophenone Triethylamine Calcium hydride Toluene Calcium hydride All the reactions were carried out under an atmosphere of nitrogen or argon in glassware dried. Chromatography on thin films (0.2Smm) was carried out on plates of glass covered with Merck 60F-250 silica gel. The plates were revealed under a ultraviolet lamp and by soaking in an ammonium molybdate solution serum then heated on a hot plate. Flash chromatographies have been performed with Merck Kieselgel silica gel (230-240 mesh).
The infrared spectra were performed using a Perkin- spectrometer Elmer 1600 FT-IR. Proton nuclear magnetic resonance (H NMR) spectra carbon (NMR
13C), fluorine (19F NMR) and selective decoupling were carried out at using a Bruker AC-300 spectrometer at 25 ° C. Chemical shifts are reported in part per million (ppm). Chloroform has been used as an internal reference for the proton (7.26 ppm) and for carbon (77.00 ppm). High resolution mass spectra and bass resolution were recorded with a VG Micromass ZAB-2F polarimeter. The points of were measured with a Büchi M-50 instrument and were not corrected.
The analysis crystallographic was performed using an Enraf Nonius diffractometer CAD-4.
The following abbreviations have been used for the interpretation of ghosts M 'Ionized mass PF, Melting Point 1 H NMR Proton nuclear magnetic resonance RMN'3C Nuclear magnetic carbon resonance EMN'9F Nuclear magnetic resonance of fluorine SM Mass spectrum SMHR High resolution mass spectrum d Doublet dd Doublet of doublet ddd doublet doublet doublet dt triplet doublet dq Quadruple doublet Byte q Quadruplet quint Quintuplet :, Singlet td doublet triplet Note: Due to the coupling between carbon and fluorine, there are a number of higher peak, in 13C NMR spectra, that the number of carbon on the molecules. This number is indicated in parenthesis after each NMR ~ 3C for the molecules concerned.

Operational modes Olefin 27 O
www Me0 I s% ~~ Me0 I ~ ' Ä a solution of ketone 21 (20 g, 113 mmol) in tetrahydrofuran (500 mL) at -78 ° C
aluminum and lithium hydride (2.2 g, 57 mmol) was added. The mixture was agitated at-78 ° C for 1 h and sodium sulfate decahydrate was added. The mixture was stirred for 1 h at room temperature and the precipitate was filtered. The solvent has been evaporated and the raw alcohol obtained was dissolved in benzene. Acid p-toluenesulfonic has been added (215 mg, 1.13 mmol) and the mixture was stirred at reflux with a dean-strak for 1.5 h. A
aqueous saturated potassium bicarbonate solution was added and the phase aqueous was extracted with ethyl acetate. The combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was filtered through silica and rinsed with 30% ethyl acetate in hexane to give the 27 olefin shape of a colorless oil (19 g, 100%).
Raw aluol Crude formula: Cl, HlaO2.
1 H-NMR (300 MHz, CDCl3), 8 (ppm): 7.34 (1H, d, J = 8.5 Hz, Ph), 6.77 (1H, dd, J = 8.5 Hz and J = 2.5 Hz, Ph), 6.63 (1 H, d, J = 2. S Hz, Ph), 4.75 (1 H, solid, CHOH), 3.79 (3H, s, ArOCH3), 2.84-2.65 (2H, t, PhCH CH?), 2.03-1.70 (4H, m, PhCH2CH Ces), 1.60 (1H, solid, CHOH) Olefin 27 Gross formula: C11Hi20.
1RMN'H (300 MHz, CDC13), 8 (ppm): 6.96 (1H, d, J = 9.1 Hz, Ph), 6.71-6.68 (2H, m, Ph), 6.42 (1H, dt, J = 9.3 Hz and J = 1.9 Hz, PhCH = CHCH2), 5.90 (1H, dt, J = 9.3 Hz and J = 4.4 Hz, PhCH = CHCHZ), 3.80 (3H, s, ArOCH), 2.78 (2H, t, J = 8.2 Hz, PhCH CH2), 2.33-2.26 (2H, m, PhCH = CHCH).
13C NMR (75 MHz, CDCI3), 8 (ppm): 158.53, 137.08, 127.32, 127.14, 126.78, 125.81, 113.79, 111.02, 55.13, 27.97, 22.96.
1R (CHC13), v (crri '): 3030, 2933, 2830, 1606, 1569, 1500, 1464, 1428, 1253 MS (m / e): 160 M +.
SMHR: calculated for C1 ~ H16O3 (M ~: 160.0888, found: 160.0887 ~ 0.0005.
Diol 20 _ ~ OH
/ 'Me0 I / OH
me0 Ä a solution of olefin 27 (26.0 g, 160.3 mmol) in methanol (2 L) at -78 ° C was bulIe of ozone for 1.5 h. Nitrogen was bubbled for 15 min and the borohydride sodium (12.1 g, 319.9 mmol) was added. The mixture was stirred for 2 h then from sodium borohydride (6.1 g, 160.0 mmol) has been added. The mixture was agitated for 1 hour then the solvent was evaporated. The residue was dissolved in the tetrahydrofuran (1 L) and one 2.5 M aqueous sodium hydroxide solution (330 mL) and peroxide hydrogen 30%
(230 mL) have been added. The mixture was stirred at room temperature All night long. A
10% aqueous sodium thiosulfate solution was added and the phase aqueous has been extracted with chloroform. The combined organic phases were washed with a solution aqueous saturated sodium bicarbonate and with water. The organic phase was dried with magnesium sulfate, filtered and evaporated to give the diol 20 sous shape of a white solid (27.1 g, 86%).

Gross formula: C11Hi603 ~
1H NMR (300 MHz, CDCl3), 8 (ppm): 7.22 (1H, d, J = 8.3 Hz, Ph), 6.78 (1H, d, J = 2.7 Hz, Ph), 6.72 (1H, dd, J = 8.3 Hz and J = 2.7 Hz, Ph), 4.63 (2H, s, ArCH OH), 3.80 (3H, s, ArOCH), 3.57 (2H, t, J = 5.9 Hz, CHZCH OH), 2.82 (2H, t, J = 7.3 Hz, ArCH
CH2), 2.49 (2H, solid, ArCH20H and CH2CH20H ~, I .93 (2H, quint, J = 6.1 Hz, CH2CH CH2).
13C NMR (75 MHz, CDC13), 8 (ppm): 159.34, 142.05, 131.06, 130.80, 115.02, 110.88, 62.41, 60.74, 55.17, 33.63, 27.26.
IR (CHCl3), v (crri l): 3327, 2939, 2875, 1610, 1580, 1500, 1460, 1256, 1160, 111 l, 1055, 1031, 942, 819.
5M (m / e): 196 M +.
SMHR: calculated for C11Hi60s (M +): 196.1099, found: 196.1108 t 0.0006.
Hydrogyaldehyde 32 O
OH ~ H
Me0 I ~ OH ~ I ~ OH
me0 Ä a solution of diol 20 (9.5 g, 48.5 mmol) in a mixture of pentane and dichloromethane 4: 1 (400 mL) was added manganese (IV) oxide (2I.0 g, 242.5 mmol) and the mixture was stirred for 30 min. Manganese (IV) oxide a been added per 21.0 g serving every 30 min until reaction is complete complete. Then the mixture was filtered on silica and the solvent was evaporated under pressure reduced to give crude hydroxyaldehyde 32 in the form of a yellow oil (8.0 g, 85%).
Raw formula: CllHia03.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 10.06 (1H, s, CHO), 7.78 (1H, d, J = 8.6 Hz, Ph), 6.89 (1H, dd, J = 8.6 Hz and J = 2.5 Hz, Ph), 6.82 (1H, d, 2.5 Hz, Ph), 3.90 (3H, s, ArOCH), 3.68 (2H, t, J ~ 6.1 Hz, CHZCH OH), 3.15 (2H, t, J = 7.8 Hz, ArCH CH2), 2.06-1. $ 5 (3H, m, CH2CH CHZOH ~.
'3C NMR (75 MHz, CDC13), ô (ppm): 191.54, 163.85, 147.53, 135.56, 127.29, 116.40, 111.68, 61.52, 55.44, 34.50, 28.93.
IR (CHC13), v (cm- '): 3414, 2942, 2872, 1682, 1599, 1566, 1496, 1463, 1431, 1290, 1251, 1121, 105, 1057, 1025, 813.
5M (m / e): 194 M +.
SMHR: calculated for C ~ 1H, 403 (M '~: 194.0943, found: 194.0948 ~ 0.0006.
Silyl Ether 34 OO
WHWH.
Me0 I ~ OH MQO I ~ OTBDMS

Ä a solution of hydroxyaldehyde 32 (8.7 g, 44.8 mmol) in the tetrahydrofuran (200 mL) was added imidazole (7.6 g, 112 mmol) and t- chloride butyldimethylsilyl (10.1 g, 67.2 mmol). The mixture was stirred at room temperature for 45 min then one saturated aqueous ammonium chloride solution was added. The sentence aqueous was extracted with ethyl acetate and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained has been purified through flash chromatography (10% ethyl acetate, 90% hexane) to give aldehyde 34 p form of a pale yellow oil (13.8 g, 100%).
Raw formula: CI ~ HZgO3Sl.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 10.14 (1H, s, CHO), 7.80 (1H, d, J = 8.6 Hz, Ph), 6.85 (1H, dd, J = 8.6 Hz and J = 2.6 Hz, Ph), 6.77 (1H, d, 2.6 Hz, Ph), 3.87 (3H, s, ArOCH), 3.68 (2H, t, J = 6.1 Hz, CH2CH OTBDMS), 3.07 (2H, dd, J = 7.9 Hz and J = 6.1 Hz, ArCH CHZ), 1.84 (2H, dt, J = 9.5 Hz and J = 6.1 Hz, CH2CH CH20TBDMS), 0.91 (9H, s, If-C (CH) 3), 0.06 (6H, s, Si (CH) 2).

13C NMR (75 MHz, CDC13), 8 (ppm): 190.75, 163.7, 147.8, 134.31, 127.5, 116.18, 111.71, 62.19, 55.43, 34.84, 29.95, 25.95, 18.3, -5.3.
1R (CHCl3), v (crri '): 2959, 2929, 2856, 2734, 1689, 1600, 1463, 1225, 1102, 1029, 836.
5M (m / e): 251 (M-C4H9) +.
SMHR: calculated for C13H19 ~ 3s1 (M-CçH9) +: 251.1103, found: 251.1108 ~
0.0007.
Amide 35 O
No me OI ~ Me ~ H
Me0 I ~ OTBDMS Me0 I ~ OTBDMS

Ä a suspension of 60% sodium hydride in oil (1.8 g, 45.6 mmol) in ether (50 mL) at 0 ° C was added phosphonate 30 (10.9 g, 45.6 mmol). The resulting mixture has been stirred at 0 ° C for 30 min. Aldehyde 34 (9.35 g, 30.4 mmol) was added by cannula and the resulting mixture was stirred at room temperature for 30 min. A
solution aqueous saturated ammonium chloride was added and the aqueous phase was extracted with ethyl acetate. The combined organic phases were dried with sulfate magnesium, filtered and evaporated under reduced pressure. The residue obtained was purified by flash chromatography (40% ethyl acetate, 60% hexane) to give the amide 35 cents form of a pale yellow oil (10.1 g, 85%).
Gross formula: C2 ~ H340aNSi.
1H NMR (300 MHz, CDCl3), 8 {ppm): 8.00 (1H, d, J = 15.6 Hz, CH = CHPh), 7.61 (1H, d, J
= 9.4 Hz, Ph), 6.88 (1H, d, J = 15.6 Hz, CH = CHPh), 6.79-6.77 (2H, m, Ph), 3.84 (3H, s, ArOCH), 3.77 (3H, s, NOCH), 3.66 (2H, t, J = 6.3 Hz, CHZCH OTBDMS), 3.32 (3H, s, NCH), 2.84 (2H, dd, J = 7.9 Hz and J = 6.2 Hz, ArCH CHZ), 1.82 (2H, dt, J =
7.9 Hz and J = 6.3 Hz, CH2CH CH2), 0.93 (9H, s, SiC (CH) 3), 0.08 (6H, s, Si (CH) 2).

13C NMR (75 MHz, CDC13), 8 (ppm): 167.33, 160.76, 143.87, 140.38, 128.07, 126.41, 115.02, 114.76, 112.17, 62.22, 61.77, 55.23, 34.27, 32.46, 29.68, 25.99, 18.32, -5.25.
IR (CHC13), v (cm 1): 2953, 2938, 2856, 1656, 1600, 1496, 1463, 1378, 1260, 1097, 1006, 837, 777.
MS (m / e): 336 (M-C4H9) +.
SMHR: calculated for C1 ~ H2504NSi (M-C4H9) +: 336.1631, found: 336.1634 ~
0.0010.
Alcohol 33 OO
N'OMe N.OMe Me I 'Me -OTBDMS I / OH
Me0 Me0 Ä a solution of silylated ether 34 (10.0 g, 50.8 mmol) in tetrahydrofuran (200 mL) in summer added a 1M solution of tetrabutylammonium fluoride in the tetrahydrofuran (50.8 mL, 50.8 mmol). The mixture was stirred at room temperature for 30 min and the solvent was evaporated. The residue obtained was purified by chromatography lightning (60%
acetone, 40% hexane) to give alcohol 33 as a pale yellow oil (7.2 g, 100%).
Gross formula: ClSHZiOaN.
1H NMR (300 MHz, CDCl3), 8 (ppm): 8.01 (1H, d, J = 15.6 Hz, CH = CHPh), 7.61 (1H, d, J
= 9.2 Hz, Ph), 6.87 (1H, d, J = 15.6 Hz, CH = CHPh), 6.79-6.76 (2H, m, Ph), 3.82 (3H, s, ArOCH), 3.76 (3H, s, NOCH), 3.68 (2H, t, J = 6.3 Hz, CHZCH OH), 3.30 (3H, s, NCH), 2.87 (2H, dd, J = 8.0 Hz and J = 6.4 Hz, ArCH CHZ), 1.92 (1H, solid, CH20H), 1.85 (2H, dt, J = 8.0 Hz and J = 6.3 Hz, CHZCH CHZ).
'3C NMR (75 MHz, CDCl3), 8 (ppm): 167.24, 160.76, 144.03, 140.42, 128.88, 125.82, 114.89, 114.05, 112.12, 61.64, 61.62, 54.98, 34.34, 32.27, 29.75.

IR (CHCl3), v (cm ''): 3414, 2939, 2874, 1646, 1596, 1496, 1461, 1381, 1308, 1257, 1002, 813, 731.
SM (m / e): 279 M'-.
SMHR: calculated for C, SH ~ 04N (MH ~: 280.1549, found: 280.1544 t 0.0008.
Aldehyde 36 O
.OMe .OMe N'Me Me Me0 I ~ OH H

Ä a solution of dimethyl sulfoxide (460 pL, 6.48 mmol) in the dichloromethane (20 mL) at -78 ° C was added oxalyl chloride (377 wL, 4.32 mmol) slowly followed by alcohol 33 (603 mg, 2.16 mmol) drop by drop. The resulting mixture was stirred at -78 ° C for 45 min and triethylamine (1.2 mL, 8.64 mmol) was added. The solution has been agitated at the room temperature for 30 min and an aqueous chloride solution ammonium saturated has been added. The aqueous phase was extracted with dichloromethane and the phases combined organic were dried with magnesium sulfate, filtered and evaporated. The residue was filtered through silica and the solvent was evaporated to give raw aldehyde 36 under form of a pale yellow oil (565 mg, 94%).
Gross formula: C, SH190aN.
1 H NMR (300 MHz, CDC13), 8 (ppm): 9.79 (1H, s, CHO), 7.92 (1H, d, J = 15.5 Hz, CH = CHPh), 7.59 (1H, d, J = 8.6 Hz, Ph), 6.88 (1H, d, J = 15.5 Hz, CH = CHPh), 6.79 (1H, dd, J = 8.5 Hz and J = 2.7 Hz, Ph), 6.75 (1H, d, J = 2.7 Hz, Ph), 3.81 (3H, s, ArOCH), 3.76 (3H, s, NOCH), 3.30 (3H, s, NCH), 3.10 (2H, t, J = 7.6 Hz, ArCH CH2), 2.75 (2H, t, J
= 7.6 Hz, CHZCI- ~ I CHO).
13C NMR (75 MHz, CDC13), 8 (ppm): 200.77, 167.02, 160.83, 141.87, 139.61, 128.35, 126.15, 115.28, 115.02, 112.57, 61.83, 55.24, 44.89, 32.40, 25.69.

IR (CHC13), v (crri '): 2940, 2836, 2725, 1727, 1653, 1601, 1497, 1378, 1306, 1256, 1173, 1096, 1048, 1001, 815.
MS (m / e): 277 M +.
SMHR: calculated for ClsHaoOaN (MH ~: 278.1392, found: 278.1397 ~ 0.0008.
Ester a, ~ 13-unsaturated 31 .OMe ~ .OMe 'Me ~ N ~ Me -If H

Ä a solution of fluorophosphonate 28 (269 mg, 1.11 mmol) in the tetrahydrofuran (20 mL) at -78 ° C was added a 1.40 M solution in n-butyl hexane lithium (793 ~ I ,, 1.11 mmol). The solution was stirred at -78 ° C for 10 min and aldehyde 36 (282 mg, 1.01 mmol) was added by cannula. The mixture was stirred at -78 ° C for 30 min then at 0 ° C
for 30 min. A 1N aqueous hydrochloric acid solution was added and the phase aqueous was extracted with ethyl acetate. Organic phases gathered were dried with magnesium sulfate, filtered and evaporated under pressure scaled down. The residue obtained was purified by flash chromatography (50% ethyl acetate, 50%
hexane) for give the ester a, ~ unsaturated 31 in the form of a yellow oil (323 mg, 88%).
Raw formula: Cl9HzaOsNF.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.97 (1H, d, J = 15.5 Hz, CH = CHPh), 7.60 (1H, d, J
= 8.6 Hz, Ph), 6.87 (1H, d, J = 15.5 Hz, CH = CHPh), 6.79 (1H, dd, J = 8.6 Hz and J = 2.7 Hz, Ph), 6.75 (1H, d, J = 2.7 Hz, Ph), 5.92 (1H, dt, J = 21.2 Hz and J = 7.7 Hz, CH = CFC02Et) 4.27 (2H, q, J = 7.1 Hz, C02CH CH3), 3.83 (3H, s, ArOCH), 3.76 (3H, s, NOCH
), 3.30 (3H, s, NCH), 2.94-2.79 (4H, m, ArCH CH), 1.32 (3H, t, J = 7.1 Hz, C02CHZCH
).

13C NMR (75 MHz, CDC13), 8 (ppm): 167.10, 160.76, 149.02, 145.90, 142.13, 139.93, 128.21, 126.34, 121.81, 121.55, 115.15, 115.02, 112.50, 61.77, 61.32, 55.18, 32.51, 32.42, 26.87, 14.02. (1 pic +) IR (CHC13), v (crri l): 2971, 2940, 1730, 1655, 1601, 1497, 1462, 1377, 1256, 1181. 1097, 1037, 1005, 858, 816, 777.
5M (m / e): 365 M +.
SMHR: calculated for C19H24OSNF (M ~: 365.1638, found: 365.1633 + 0.0011.
Hydroxyaldehyde 37 OO
.OMe \ N ~ Me ~ H
FI ~ F
me0 C02And Me0 OH

Ä a solution of the amide-ester 31 (6.7 g, 18.4 mmol) in tetrahydrofuran (150 mL) at-78 ° C was added aluminum and lithium hydride (1.05 g, 27.6 mmol). The mixture was stirred at -78 ° C for 1.5 h and at 0 ° C for 45 min. Of acetone was added followed by sodium sulfate decahydrate and the resulting mixture was stirred for 1 h then filtered. The solvent was evaporated and the residue was purified by flash chromatography (30% acetone, 70% hexane) to give hydroxyaldehyde 37 as a yellow solid (2.15 g, 44%).
Gross formula: C ~ SH1 ~ 03F.
1H NMR (300 MHz, CDCl3), 8 (ppm): 9.69 (1H, d, J = 7.6 Hz, CHO), 7.70 (1H, d, J = 15.7 Hz, CH = CHPh), 7.62 (1 H, d, J = 8.7 Hz, Ph), 6.84 (1 H, dd, J = 8.7 Hz and J
= 2.7 Hz, Ph), 6.74 (1 H, d, J = 2.7 Hz, Ph), 6.62 (1 H, dd, J = 15. 7 Hz and J = 7.6 Hz, CH = CHPh), 5.24 (1 H, dt, J = 21.3 Hz and J = 7.5 Hz, CH = CFCHZOH), 4.05 (2H, d, J = 21.3 Hz, CH OH), 3.85 (3H, s, ArOCH), 2.87 (2H, t, J = 7.5 Hz, ArCH CH2), 2.33 (2H, q, J = 7.5 Hz, CH CH = CFCHZOH), 1.58 (1H, solid, CH20H).

MS (m / e): 264 M +.
SMHR: calculated for C15H1 ~ 03F (M ~: 264.1162, found: 264.1159 ~ 0.0008.
Chloride 38 M

Ä a solution of alcohol 37 (2.15 g, 8.14 mmol) in tetrahydrofuran (100 mL) at --40 ° C
were added triphenylphosphine (2.3 g, 8.95 mmol) and hexachloroacetone (1.2 mL, 8.14 mmol). The mixture was stirred at -40 ° C for 30 min then the solvent was been evaporated. The residue obtained was purified by flash chromatography (30% ethyl acetate, 70% hexane) to give chloride 38 as a pale yellow solid (1.78 g, 77%).
Raw formula: CISHis0zFCl.
1 H NMR (300 MHz, CDC13), 8 (ppm): 9.71 (1 H, d, J = 7.7 Hz, CHO), 7.70 (1 H, d, J = 15.6 Hz, CH = CHPh), 7.63 (1 H, d, J = 8.7 Hz, Ph), 6.84 (1 H, dd, J = 8.7 Hz and J
= 2.7 Hz, Ph), 6.75 (1H, d, J = 2.7 Hz, Ph), 6.62 (1H, dd, J = 15.6 Hz and J = 7.7 Hz, CH = CHPh), 5.32 (1H, dt, J = 18.5 Hz and J = 8.3 Hz, CH = CFCH2C1), 4.00 (2H, d, J = 21.5 Hz, CH Cl), 3.85 (3H, s, ArOCH), 2.89 (2H, t, J = 7.5 Hz, ArCH CH2), 2.33 (2H, q, J = 7.7 Hz, CH
CH = CFCHZCI).
MS (m / e): 282 M +, 247 (M-Cl) +.
SMHR: calculated for C15Hi60zFC1 (M ~: 282.0853, found: 282.0828 ~ 0.0008.

Alcohol 39 OH
F
M Me0 THIS

to a solution of aldehyde 38 (1.78 g, 6.30 mmol) in methanol (60 mL) a added on sodium borohydride (477 mg, 12.6 mmol) and the solution was stirred temperature ambient for 2 h. A saturated aqueous solution of sodium bicarbonate has been added and the aqueous phase was extracted with ethyl acetate. The phases organic together have been dried with magnesium sulfate, filtered and concentrated under reduced pressure. The residue obtained was purified by flash chromatography (40% ethyl acetate, 60% hexane) to give alcohol 39 as a white solid (1.51 g, 54%).
Raw formula: CISHi802FC1.
Mp: 47-49 ° C
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.41 (1H, d, J = 8.6 Hz, Ph), 6.80-6.74 (2H, m, CH = CHPh and Ph), 6.65 (1 H, d, J = 2.7 Hz, Ph), 6.18 (1 H, dt, J = 15.6 Hz and J = 5.8 Hz, CH = CHPh), 5.31 (1H, dt, J = 18.8 Hz and J = 8.3 Hz, CH = CFCH2CI), 4.33 (2H, dd, J = 5.8 Hz and J = 1.5 Hz, CH OH), 3.99 (2H, d, J = 21.4 Hz, CH Cl), 3.80 (3H, s, ArOCH), 2.75 (2H, t, J = 7.3 Hz, ArCH CH2), 2.29 (2H, q, J = 7.9 Hz, CH CH = CFCH2Cl), 1.56 (1H, massive, CHZOH ~.
'3C NMR (75 MHz, CDCl3), 8 (ppm): 159.14, 156.79, 139.51, 128.77, 128.14, 127.84, 127.59, 115.06, 112.19, 110.28, 110.01, 63.83, 55.25, 37.70, 37.27, 33.30, 27.06, 26.96 (3 more peaks).
IR (CHC13), v (cm '): 3373, 2942, 2838, 1734, 1695, 1494, 1257, 1084, 1041, 968, 834, 723.
5M (m / e): 284 M +.
SMHR: calculated for CISH ~ gOZFCI (M +): 284.0979, found: 284.0974 ~ 0.0008.

Silyl Ether 40 OH I OTBDMS
~ F _- ~. I ~ F
Me0 ~ C ~ Me0 ~ C

Ä a solution of alcohol 39 (1.51 g, 5.31 mmol) in tetrahydrofuran (50 rnL) have been added imidazole (903 mg, 13.3 rnmol) and t- chloride butyldimethylsilane (1.20 g, 7.97 mmol). The mixture was stirred at room temperature for 2 h and a aqueous solution 0.5 M citric acid was added. The aqueous phase was extracted with dichlorornéthane and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (10%
ethyl acetate, 90% hexane) to give silylated ether 40 as a pale yellow oil (1.50 g, 71%).
Gross formula: C21H3a02FC1Si.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.42 (1H, d, J = 8.5 Hz, Ph), 6.79-6.74 (2H, m, CH = CHPh and Ph), 6.66 (1 H, d, J = 2.7 Hz, Ph), 6.11 (1 H, dt, J = 15.6 Hz and J = 5.0 Hz, CH = CHPh), 5.31 (1H, dt, J = 18.8 Hz and J = 8.3 Hz, CH = CFCHZCI), 4.38 (2H, dd, J = S.0 Hz and J = 1.8 Hz, CH OTBDMS), 3.98 (2H, d, J = 21.4 Hz, CH Cl), 3.82 (3H, s, ArOCH), 2.77 (2H, t, J = 7.8 Hz, ArCH CHZ), 2.29 (2H, q, J = 7.8 Hz, CH CH = CFCHZCI), 0.96 (9H, s, Si-C (CH) 3), 0.13 (6H, s, Si (CH) 2).
IR (CHC13), v (cm ''): 2951, 2857, 1608, 1496, 1464, 1255, 1121, 1094, 966, 836, 778, 725.
MS (m / e): 398 M +.
SMHR: calculated for CZ1H3202FC1Si (M ~: 398.1844, found: 398.1849 ~
0.0012.

, ~ Keto ester 41 OTBDMS OTBDMS
i F ~ I ~ F
Me0 ~ C ~ Me0 ~ OMe Solution 1: Ä a suspension of 60% sodium hydride in oil (1.1 g, 28.2 mmol) in tetrahydrofuran (20 mL) at 0 ° C was added acetoacetate methyl (2.8 mL, 26.3 mmol) and the mixture was stirred at 0 ° C for 30 min. A solution of n-butyl lithium 1.4 M in hexane (18.8 mL, 26.3 mmol) was added to this mixture at 0 ° C and the unrest was continued for 30 min.
Solution 2: Ä a solution of chloride 40 (1.50 g, 3.76 mmol) in the tetrahydrofuran (20 mL) was added sodium iodide (1.7 g, 11.3 mmol) and the mixture was agitated at the room temperature for 30 min.
Solution 1 was added by cannula to solution 2 and the mixture obtained been agitated at the room temperature overnight. An aqueous chloride solution ammonium saturated was added and the aqueous phase was extracted with acetate ethyl. The phases combined organic were dried with magnesium sulfate, filtered and evaporated under reduced pressure. The residue obtained was purified by flash chromatography (20% acetate ethyl, 80% hexane) to give the / ~ keto ester 41 in the form of an oil pale yellow (1.36 g, 76%).
Gross formula: C26H3905FS ~.
1H NMR (300 MHz, CDC13), 8 (ppm): 12.05 (s, RC (O) CH = C (OH) R ') 7.38 (1H, d, J =
8.6 Hz, Ph), 6.79-6.71 (2H, m, CH = CHPh and Ph), 6.66 (1H, d, J = 2.7 Hz, Ph), 6.06 (1H, dt, J =
15.6 Hz and J = 4.9 Hz, CH = CHPh), 5.07 (1H, dt, J = 21.7 Hz and J = 8.1 Hz, CH = CFR), 4.35 (2H, dd, J = 4.9 Hz and J = 1.7 Hz, CH OTBDMS), 3.80 (3H, s, AxOCH), 3.73 (3H, s C02CH), 3.40 (2H, s, C (O) CH C (O)), 2.70 (2H, t, J = 7.4 Hz, ArCH CHZ), 2.51-2.47 (2H, m, CHZCH C (O)), 2.42-2.30 (2H, m, CH CHZC (O)), 2.22 (2H, q, J = 7.6 Hz, CH
CH = CFR) 0.95 (9H, s, Si-C (CH) 3), 0.11 (6H, s, Si (CH3) 2).
'3C NMR (75 MHz, CDC13), 8 (ppm): 200.91, 160.43, 158.83, 157.16, 139.95, 129.06, 128.61, 127.57, 126.05, 115.15, 112.03, 105.51, 105.22, 63.91, 55.13, 52.24, 48.80, 39.08, 33.53, 26.77, 26.64, 25.94, 22.00, 21.62, 18.37, -5.16 (3 more peaks).
IR (CHCI3), v (crri l): 2952, 2857, 1748, 1720, 1609, 1496, 1462, 1254, 1090, 966, 537, 778.
5M (m / e): 478 M +.
SMHR: calculated for C26H3905FSi (M +): 478.2551, found: 478.2536 ~ 0.0014.
Ester a ,, l ~ -unsaturated 44 O
OI 'OEt -, w Me0 I ~ OTBDMS Me0 I ~ OTBDMS

Ä a suspension of 60% sodium hydride in oil (2.5 g, 61.8 mmol) in the tetrahydrofuran (350 mL) was added triethyl phosphonoacetate (12.3 mL, 61.8 mmol). The resulting mixture was stirred at room temperature for 15 min. A
aldehyde solution 34 (12.7 g, 41.2 mmol) in tetrahydrofuran (50 mL) has been added per cannula in the anion solution. The mixture was stirred ambient temperature for 20 min and a saturated aqueous ammonium chloride solution was added. The aqueous phase was extracted with ethyl acetate and the phases organic together were dried with magnesium sulfate, filtered and evaporated. The residue obtained was filtered on silica then rinsed with 10% ethyl acetate in hexane to give the ester has, /. ~ unsaturated 44 in the form of a pale yellow oil (15.2 g, 97%).

Gross formula: C21H3aOaSi.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.95 (1H, d, J = 15.8 Hz, PhCH = CH), 7.56 (1H, d, J
= 9.2 Hz, Ph), 6.76-6.73 (2H, m, Ph), 6.28 (1 H, d, J = 15.8 Hz, PhCH = CI- ~ i, 4.27 (2H, q, J =
7.1 Hz, C02CH CH3), 3.84 (3H, s, ArOCI- ~ I), 3.77 (2H, t, J = 6.3 Hz, CH2CH
OTBDMS) 2.81 (2H, dd, J = 9.7 Hz and J = 6.1 Hz, ArCH CHz), 1.81 (2H, dt, J = 9.7 Hz and J = 6.1 Hz, CH2CH CH20TBDMS), 1.35 (3H, t, J = 7.1, C02CH2CH), 0.93 (9H, s, SiC (CH) 3), 0.08 (6H, s, Si (CH) 2).
'3C NMR (75 MHz, CDCl3), 8 (ppm): 167.26, 161.01, 143.91, 141.48, 128.14, 125.57, 116.95, 115.15, 112.17, 62.17, 60.22, 55.16, 34.34, 29.67, 25.95, 18.30, 14.36, -5.31.
IR (CHCl3), v (cm '): 2954, 2857, 1712, 1603, 1495, 1464, 1366, 1295, 1257, 1159, 1105, 1032, 979, 837.
MS (m / e): 363 (M-CH3) +, 321 (M-C4H9) +.
SMHR: calculated for C2oH31OaSi (M-CH3) +: 363.1991, found: 363.1995 ~
0.0011.
Alcohol 45 O
AND ~ OH
Me0 I ~ OTBDMS I ~ OTBDMS
me0 Ä a solution of the ester a, ~ unsaturated 44 (18.4 g, 48.7 mmol) in the dichloromethane (450 mL) at 0 ° C was added a 1 M düsobutylaluminium hydride solution in the dichloromethane (121.8 mL, 121.8 mmol). The mixture was stirred at 0 ° C
for 1 hour and the sodium sulfate decahydrate was added. The residue obtained was filtered and then rinsed with ethyl acetate and dichloromethane. The filtrate was concentrated under reduced pressure for give the alcohol 45 in the form of a yellow oil (15.9 g, 97%).

Gross formula: C1gH3203S1.
NMR ~ H (300 MHz, CDC13), S (ppm): 7.43 (1H, d, J = 8.4 Hz, Ph), 6.83 (1H, d, J = 15.7 Hz, PhCH = CH), 6.77-6.72 (2H, m, Ph), 6.18 (1H, dt, J = 15.7 Hz and J = 6.0 Hz, PhCH = Ces, 4.32 (2H, dd, J = 6.0 Hz and J = 1.3 Hz, CH OH), 3.82 (3H, s, ArOCH), 3.67 (2H, t, J = 6.2 Hz, CH2CH OTBDMS), 2.76-2.71 (2H, m, ArCH CH2), 1.79 (2H, dt, J = 9.9 Hz and J =
6.1 Hz, CH2CH CHZOTBDMS), 1.60 (1H, solid, CH20H ,, 0.94 (9H, s, SiC (CH) 3), 0.09 (6H, s If (CH) 2).
13C NMR (75 MHz, CDC13), b (ppm): 159.13, 141.29, 128.50, 128.11, 128.06, 127.27, 114.85, 111.74, 64.06, 62.44, 55.18, 34.02, 29.76, 25.98, 18.35, -5.25.
IR (CHC13), v (cm-1): 3356, 2952, 2857, 1607, 1494, 1464, 1256, 1098, 1033, 967, 836.
MS (m / e): 336 M +.
SMHR: calculated for C ~ 9H32O3S1 (M) +: 336.2121, found: 336.2123 ~ 0.0010.
Silyl ether 46c OH ~ I OTBDPS
Me0 I ~ OTBDMS MQO I / OTBDMS

to a solution of alcohol 45 (15.9 g, 47.3 mmol) in tetrahydrofuran (400 mL) have been added imidazole (8.1 g, 118.3 mmol) and t- chloride butyldiphenylsilane (18.5 mL, 71.0 mmol). The mixture was stirred at room temperature for 1.5 h and a solution aqueous saturated ammonium chloride was added. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried with sulfate magnesium, filtered and evaporated. The residue obtained was purified by flash chromatography (5% ethyl acetate, 95% hexane) to give the disilylated compound 46c under shape of a yellow oil (25.0 g, 93%).

Gross formula: C35H50 ~ 3S12.
1H NMR (300 MHz, CDCl3), 8 (ppm): 7.74 (4H, dd, J = 7.5 Hz and J = 1.7 Hz, Ph), 7.48-7.36 (7H, m, Ph), 6.87 (1H, d, J = 15.7 Hz, PhCH = CH), 6.76-6.72 (2H, m, Ph), 6.10 (1H, dt, J =
15.7 Hz and J = 4.9 Hz, PhCH = CHI, 4.39 (2H, dd, J = 6.7 Hz and J = 1.8 Hz, CH
OTBDPS) 3.82 (3H, s, ArOCH), 3.63 (2H, t, J = 6.3 Hz, CH2CH OTBDMS), 2.71 (2H, dd, J
= 9.7 Hz and J = 7.7 Hz, ArCH CH2), 1.84-1.74 (2H, m, CH2CH CH20TBDMS), 1.58 (9H, s, SiC (CH) 3), 1.11 (9H, s, SiC (CH) 3), 0.06 (6H, s, Si (CH) 2).
'3C NMR (75 MHz, CDCl3), 8 (ppm): 158.92, 141.09, 135.59, 133.79, 129.69, 128.22, 127.73, 127.65, 127.16, 126.46, 114.86, 111.77, 64.84, 62.45, 55.23, 33.97, 29.87, 26.91, 26.02, 19.32, 19.35, -5.20.
IR (CHCl3), v (cry 1): 3048, 2934, 2858, 1607, 1495, 1466, 1254, 1106, 1062, 967, 837.
MS (m / e): 574 M +.
SMHR: calculated for C35H50 ~ 3s12 (M) +: 574.3298, found: 574.3303 t 0.0017.
Alcohol 47 OTBDPS ~ I OTBDPS
Me0 I ~ OTBDMS (r OH
me0 46c 47 Ä a solution of the disilylated compound 46c (40.3 g, 702. mmol) in 95% ethanol (500 mL) a added pyridinium p-toluenesulfonate (1.8 g, 7.02 mmol) and mixture was stirred overnight at room temperature. An aqueous bicarbonate solution sodium saturated was added and the aqueous phase was extracted with acetate ethyl. The phases combined organic were dried with magnesium sulfate, filtered and concentrated under reduced pressure. The residue obtained was purified by flash chromatography (50% acetate ethyl, 50% hexane) to give alcohol 47 as a yellow oil pale (31.5 g, 98%).

Gross formula: C29H36 ~ 3s1.
1 H NMR (300 MHz, CDCI3), 8 (ppm): 7.75 (4H, dd, J = 7.5 Hz and J = 1.7 Hz, Ph), 7.48-7.38 (7H, m, Ph), 6.89 (1H, dt, J = 15.6 Hz and J = 1.7 Hz, PhCH = CH), 6.78-6.73 (2H, m, Ph), 6.11 (1 H, dt, J = 15.6 Hz and J = 4.7 Hz, PhCH = Ces, 4.41 (2H, dd, J = 4.7 Hz and J
= 1.9 Hz, CH OTBDPS), 3.82 (3H, s, ArOCH), 3.65 (2H, t, J = 6.3 Hz, CH2CH OH), 2.75 (2H, dd, J
= 7.9 Hz and J = 6.2 Hz, ArCH CHz), 1.89-1.80 (2H, m, CH2CH CH2), 1.61 (1H, massive, CH20 ~, 1.1 l (9H, s, SiC (CH) 3).
13C NMR (75 MHz, CDCI3), 8 (ppm): 158.94, 140.71, 135.56, 133.72, 129.71, 128.63, 128.35, 127.73, 127.25, 126.29, 114.84, 111.85, 64.68, 62.19, 55.25, 33.78, 29.74, 26.86,

19.32.
IR (CHC13), v (cm 1): 3390, 3048, 2937, 2859, 1607, 1496, 1465, 1380, 1256, 1202, 1110, 1057, 967.
5M (m / e): 403 (M-C4H9) +.
SMHR: calculated for C25H2 ~ 03Si (M-C4H9) +: 403.1729, found: 403.1734 ~
0.0012.
Aldehyde 49 OTBDPS ~ ~ OTBDPS
\ \
Me0 I ~ OH Me0 I ~ H

Ä a solution of dimethyl sulfoxide (14.6 mL, 205.5 mmol) in the dichloromethane (700 mL) at -78 ° C was added oxalyl chloride (12.0 mL, 137.0 mmol) slowly followed by alcohol 47 (31.5 g, 68.5 mmol) drop by drop. The resulting mixture was stirred at -78 ° C
for 45 min and triethylamine (38.5 mL, 274.0 mmol) was added. The solution was stirred at room temperature for 1 h and an aqueous solution of chloride saturated ammonium was added. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was filtered through silica and the solvent was evaporated to give crude aldehyde 49 in the form of a yellow oil (31.5 mg, 100%).
Gross formula: C29H34 ~ 3S1.
1 H NMR (300 MHz, CDC13), 8 (ppm): 9.78 (1H, s, CHO), 7.73 (4H, dd, J = 7.5 Hz and J = 1.6 Hz, Ph), 7.49-7.38 (7H, m, Ph), 6.83 (1H, dt, J = 15.6 Hz and J = 1.8 Hz, PhCH = CH), 6.77 (1 H, dd, J = 5.8 Hz and J = 2.8 Hz, Ph), 6.71 (1 H, d, J = 2.8 Hz, Ph), 6.11 (1 H, dt, J = 15.5 Hz and J = 4.7 Hz, PhCH = CHI, 4.41 (2H, dd, J = 4.7 Hz and J = 1.9 Hz, CH OTBDPS), 3.82 (3H, s, ArOCH), 2.98 (2H, dt, J = 7.4 Hz, ArCH CH2), 2.71 (2H, t, J = 7.24 Hz, CH2CH
CHO) 1.11 (9H, s, SiC (CH) 3).
'3C NMR (75 MHz, CDC13), 8 (ppm): 201.26, 139.06, 135.56, 133.68, 129.76, 129.03, 128.54, 127.83, 127.76, 127.67, 127.51, 125.70, 114.77, 112.26, 64.57, 55.27, 44.74, 26.87, 25.97, 19.33.
IR (CHCl3), v (cm- '): 3078, 2935, 2857, 2721, 1925, 1607, 1496, 1428, 1256, 1110, 967.
5M (m / e): 458 M +, 401 (M-C4H9) +.
SMHR: calculated for C29H34O3Si (M ~: 458.2277, found: 458.2282 ~ 0.0014.
Ester a, ~ i-unsaturated 50 OTBDPS I OTBDPS
F
Me0 I ~ H Me0 I ~ CO And 49 p 50 Ä a solution of fluorophosphonate 28 (10.2 g, 42.3 mmol) in the tetrahydrofuran (700 mL) at -78 ° C was added a 1.40 M solution in n-butyl hexane lithium (32.0 mL, 42.3 mmol). The solution was stirred at -78 ° C for 10 min and aldehyde 49 (12.9 g, 28.2 mmol) in tetrahydrofuran was added by cannula. The mixture was stirred at -78 ° C
for 45 min. A saturated aqueous ammonium chloride solution was added and the aqueous phase was extracted with ethyl acetate. Organic phases gathered were dried with magnesium sulfate, filtered and evaporated under pressure scaled down. The residue obtained was purified by flash chromatography (10% ethyl acetate, 90%
hexane) for give the α, 3-unsaturated ester 50 as a colorless oil (14.3 g, 93%
Gross formula: C33H39 ~ 4S1F.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.75 (4H, dd, J = 7.5 Hz and J = 1.6 Hz, Ph), 7.46-7.37 (7H, m, Ph), 6.89 (1H, dt, J = 15.6 Hz and J = 1.7 Hz, PhCH = CH), 6.78 (1H, dd, J = 8.5 Hz and J
= 2.7 Hz, Ph), 6. 72 (1 H, d, J = 2.7 Hz, Ph), 6.12 (1 H, dt, J = 15.5 Hz and J = 4. 8 Hz, PhCH = CH), 5.97-5.84 (1H, m, CH2CH = CFCOZEt), 4.42 (2H, dd, J = 4.8 Hz and J =
1.8 Hz, CH OTBDPS), 4.26 (2H, q, J = 7.1 Hz, C02CH CH3), 3.83 (3H, s, ArOCH), 2.80-2.79 (4H, m, ArCH CH), 1.33 (3H, t, J = 7.1 Hz, COZCHzCH), 1.12 (9H, s, SiC (CH) 3).
'3C NMR (75 MHz, CDCl3), 8 (ppm): 158.96, 148.99, 145.65, 139.41, 135.56, 134.84, 133.74, 129.72, 129.66, 128.71, 128.67, 127.73 127.36, 126.02, 122.35, 122.10, 114.87, 112.16, 64.68, 61.36, 55.25, 32.83, 26.87.26.64, 26.59, 19.33, 14.15 (4 peaks Moreover).
IR (CHCl3), v (cm '): 3048, 2935, 2858, 1728, 1607, 1496, 1465, 1376, 1254, 1112, 1047, 967, 822.
MS (m / e): 546 M +.
SMHR: calculated for C33H39 ~ 4S1F (M ~: 546.2601, found: 546.2596 t 0.0016.
Alcohol 51 OTBDPS OTBDPS
---~ F
F
Me0 / C02And Me0 OH

Ä a solution of the a, ~ 3-unsaturated ester 50 (33.4 g, 61.2 mmol) in the tetrahydrofuran (600 mL) at 0 ° C. was added aluminum and lithium hydride (2.32 g, 61.2 mmol). The mixture was stirred at 0 ° C for 20 min and sodium sulfate decahydrate was been added. The residue obtained was filtered and then rinsed with ethyl acetate and dichloromethane. The filtrate was evaporated to give alcohol 51 as a colorless oil (30.8 g, 100%
Gross formula: C3 ~ H37O3S1F.
1H NMR (300 MHz, CDCl3), 8 (ppm): 7.77 (4H, dd, J = 7.5 Hz and J = 1.7 Hz, Ph), 7.50-7.40 (7H, m, Ph), 6.90 (1 H, dt, J = 15.6 Hz and J = 1.6 Hz, PhCH = CH), 6.79 (1 H, dd, J = 8.6 Hz and J
= 2.7Hz, Ph), 6.69 (lH, d, J = 2.7Hz, Ph), 6.14 (lH, dt, J = 15.5HzetJ = 4.6Hz, PhCH = CIT), 5.23 (1H, dt, J = 20.6 Hz and J = 8.4 Hz CH2CH = CFCH20H), 4.44 (2H, dd, J =
4.6 Hz and J = 1.9 Hz, CH OTBDPS), 3.98 (2H, d, J = 21.4 Hz, CH OH), 3.83 (3H, s ArOCH), 2.76 (2H, t, J = 7.3 Hz, ArCH CHz), 2.30 (2H, q, J = 7.6 Hz, ArCH2CH
), 1.49 (1H, solid, CH20I- ~ I, 1.15 (9H, s, SiC (CH) 3).
'3C NMR (75 MHz, CDC13), 8 (ppm): 160.58, 128.82, 157.32, 139.64, 135.55, 133.65, 129.76, 128.77, 127.76, 127.39, 125.97, 115.45, 112.15, i 07.54, 107.58, 65.00, 57.27, 56.87, 55.30, 33.68, 26.88, 26.49, 26.37, 19.34 (3 more peaks).
TR (CHC13), v (cm-1): 3433, 3047, 2936, 2858, 1735, 1607, 1496, 1429, 1253, 111 l, 967, 854.
MS (m / e): 504 M +.
SMHR: calculated for C31H3 ~ 03SiF (M '' ~: 504.2496, found: 504.2488 t 0.0015.
Chloride 52 OTBDPS ~ ~ OTBDPS
FI ~ F
Me0 ~ OH Me0 CI

Ä a solution of alcohol 51 (8.6 g, 17.1 mmol) in tetrahydrofuran (200 mL) at -40 ° C
were added triphenylphosphine (4.9 g, 18.8 mmol) and hexachloroacetone (2.6 mL, 17.1 mmol). The resulting mixture was stirred at -40 ° C for 20 min and the solvent was evaporated.

The residue obtained was purified by flash chromatography (100% hexane at 20%
acetate ethyl, 80% hexane) to give chloride 52 as a yellow oil (8.8 g, 99%
Raw formula: C31H36O2SiFCl.
1 H NMR (300 MHz, CDCl3), b (ppm): 7.75 (4H, dd, J = 7.6 Hz and J = I.6 Hz, Ph}, 7.50-7.39 (7H, m, Ph), 6.87 (1H, dt, J = 15.6 Hz and J = 1.8 Hz, PhCH = CH), 6.79 (1H, dd, J = 8.6 Hz and J
= 2.7 Hz, Ph), 6.68 (IH, d, J = 2.7 Hz, Ph), 6.13 (1 H, dt, J = 15.6 Hz and J = 4.7 Hz, PhCH = CHI, 5.31 (1 H, dt, J = 18.8 Hz and J = 8.3 Hz CHzCH = CFCHzCI), 4.44 (2H, dd, J = 4.7 Hz and J = 1.9 Hz, CH OTBDPS), 3.94 (2H, d, J = 21.4 Hz, CH Cl), 3.83 (3H, s, ArOCH), 2.76 (2H, t, J = 7.3 Hz, ArCH CH2), 2.30 (2H, q, J = 7.8 Hz, ArCH2CH), 1.13 (9H, s, SiC (CH) 3).
13C NMR (75 MHz, CDC13), 8 (ppm): 158.92, 156.83, 153.55, 139.27, 135.54, 133.67, 129.74, 128.86, 128.55, 127.74, 127.65, 127.47, 125.88, 115.14, 112.17, 110.16, 109.89, 64.60, 55.29, 37.64, 37.20, 33.39, 26.91, 26.86, 19.33 (4 more peaks).
IR (CHC13), v (cni l): 3017, 2936, 2859, 1732, 1607, 1496, 1465, 1429, 1254, 1217, 1111, 1047, 967.
MS (m / e): 520 M +.
SM (~: calculated for C31Hs602SiFC1 (M +). 522.2157, found: 522.2153 ~
0.0016.
~ 3-Keto ester 53 OTBDPS ~ OTBDPS
WFI w F
CI Me0 OMe me0 Solution 1: Ä a suspension of sodium hydride 60% in oil (592 mg, 14.8 mmol) in tetrahydrofuran (20 mL) at 0 ° C was added acetoacetate methyl (2.8 mL, 26.3 mmol) and the mixture was stirred at 0 ° C for 30 min. A solution of n-butyl lithium 1.6 M in hexane (8.6 mL, 13.8 mmol) was added to this mixture at 0 ° C
and the commotion was continued for 30 min.
Solution 2: to a solution of chloride 52 (1.03 g, 1.97 mmol) in the tetrahydrofuran (20 mL) was added sodium iodide (885 mg, S.9 mmol) and the mixture was agitated at the room temperature for 30 min.
Solution 1 was added by cannula to solution 2 and the mixture obtained been agitated at the room temperature overnight. An aqueous chloride solution ammonium saturated was added and the aqueous phase was extracted with acetate ethyl. The phases combined organic were dried with magnesium sulfate, filtered and evaporated under reduced pressure. The residue obtained was purified by flash chromatography (30% acetate ethyl, 70% hexane) to give the ~ -keto ester 53 in the form of an oil yellow (770 mg, 65%).
Gross formula: C36H43OSFSi.
1 H NMR (300 MHz, CDC13), 8 (ppm): 12.05 (RC (O) CH = C (OH) R '), 7.75 (4H, dd, J =
7.4 Hz and J = 1.7 Hz, Ph), 7.49-7.37 (7H, m, Ph), 6.92-6. 87 (1 H, m, CH = CHPh) ,. 7.77 (1 H, dd, J
= 8. S Hz and J = 2.7 Hz, Ph), 6.70 (1 H, d, J = 2.7 Hz, Ph), 6.11 (1 H, dt, J = 15.5 Hz and J = 4.6 Hz, CH = CHPh), S.O6 (1H, dt, J = 21.7 Hz and J = 8.1 Hz, CH = CFR), 4.43 (2H, dd, J = 4.6 Hz and J = 1.8 Hz, CH OTBDPS), 3.82 (3H, s, ArOCH), 3.73 (3H, s, COZCH), 3.39 (2H, s, C (O) CH ~ C (O)), 2.72 (2H, t, J = 7.3 Hz, ArCH CHZ), 2.5S-2.20 (6H, m, CHZCH
CO), CH CHaC (O)) and CH CH = CFR), 1.13 (9H, s, Si-C (CH) 3).
'3C NMR (7S MHz, CDC13), 8 (ppm): 167.36, 160.38, 158.83, 157.12, 139.99, 135.54, 133.72, 129.71, 128.70, 128.61, 127.73, 127.25, 126.01, 115.17, 112.07, 105.57, 105.28, 64.58, SS.2S, 52.35, 48.89, 39.14, 33.58, 26.86, 26.75, 26.63, 22.02, 21.65, 19.31 (3 peaks of more).
IR (CHCl3), v (crri '): 3020, 2955, 2860, 1745, 1720, 1608, 1496, 1433, 131 S, 1217, 1110, 967, 756.
5M (m / e): 602 M +.
SMHR: calculated for C36H430sFSi (M ~: 602.2864, found: 602.2870 ~ 0.0018.

Alcohol 54 OH
F
Me0 I '~ OMe to a solution of silylated enol ether 53 (1.72 g, 2.85 mmol) in Ie tetrahydrofuran (30 mL) was added a solution of 1 M tetrabutylammonium fluoride in the tetrahydrofuran (5.7 mL, 5.70 mmol) and the mixture was stirred ambient temperature for 1.5 h. The solvent was evaporated under reduced pressure and the residue obtained has been purified by flash chromatography (70% ethyl acetate, 70% hexane) to give alcohol 54 pennies form of a colorless oil (798 mg, 77%).
Raw formula: C2oH2sOsF.
NMR ~ H (300 MHz, CDC13), 8 (ppm): 7.41 (1H, d, J = 8.6 Hz, Ph), 6.83 (1H, d, J = 15.7 Hz, CH = C_HPh), 6.76 (1H, dd, J = 8.6 Hz, and J = 2.7 Hz, Ph), 6.68 (1H, d, J = 2.7 Hz, Ph), 6.18 (1 H, dt, J = 15 .6 Hz and J = 5.6 Hz, CH = CHPh), 5.10 (IH, dt, J = 2 I .6 Hz and J = 8 .2 Hz, CH = CFR), 4.34 (2H, dd, J = 5.6 Hz and J = 1.5 Hz, CH OH), 3.82 (3H, s, ArOCH), 3.76 (3H, s, C02CH), 3.46 (2H, s, C (O) CH C (O)), 2.72 (2H, t, J = 8.0 Hz, ArCH CH2), 2.63-2.58 (2H, m, CH2CH C (O)), 2.48-2.36 (2H, m, CH CH2C (O)), 2.23 (2H, q, J = 7.8 Hz, CH
CH = CFR) 1.67 (1H, massive, OH).
13C NMR (75 MHz, CDCl3), 8 (ppm): 201.71, 167.53, 160.32, 158.96, 157.05, 140.21, 128.88, 128.38, 127.45, 127.35, 115.10, 112.03, 105.68, 105.40, 63.56, 55.18, 52.39, 48.84, 39.12, 33.73, 26.93, 26.81, 21.96, 21.57 (4 more peaks).
IR (CHCl3), v (cm 1): 3425, 2949, 2867, 1744, 1716, 1608, 1496, 1441, 1372, 1254, 1084, 1008, 912, 733.
5M (m / e): 364 M +.
SMHR: calculated for C2aHzsOSF (M +): 364.1686, found: 364.1683 t 0.0011.

Pivalo ester 18 OH
OPiv F .- .. ~ ~ F
Me0 ~ OMe Me0 I ~ OMe OO 18 O Ö
Ä a solution of alcohol 54 (798 mg, 2.19 mmol) in dichloromethane (20 mL) have been added 2,6-lutidine (765 N, L,, 6.57 mmol) and chloride trimethylacetyl (405 ~, L, 3.29 mmol) and the mixture was stirred at room temperature overnight. A
aqueous solution saturated ammonium chloride was added and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried with sulfate of magnesium, filtered and evaporated. The residue obtained was purified by flash chromatography (30% ethyl acetate, 70% hexane) to give 18 pivalo tester form of an oil colorless (952 mg, 97%).
Gross formula: C25H33 ~ 6F ~
1H NMR (300 MHz, CDC13), 8 (ppm): 12.05 (RC (O) CH = C (OH) R ') 7.40 (1H, d, J =
8.6 Hz, Ph), 6.83 (1 H, d, J = 1 S. 7 Hz, CH = CHPh), 6.75 (IH, dd, J = 8.6 Hz, and J =
3.0 Hz, Ph), 6.67 (1 H, d, J = 2.7 Hz, Ph), 6.06 (IH, dt, J = 1 S. 6 Hz and J = 6.2 Hz, CH = CHPh), 5.11 (1 H, dt, J =
21.6 Hz and J = 8.I Hz, CH = CFR), 4.73 (2H, dd, J = 6.2 Hz and J = 1.3 Hz, CH
OPiv), 3.80 (3H, s, ArOCH), 3.73 (3H, s, COZCH), 3.41 (2H, s, C (O) CH C (O)), 2.71 (2H, t, J = 7.5 Hz, ArCH CH2), 2.54-2.49 (2H, m, CH2CH C (O)), 2.43-2.31 (2H, m, CH CH2C (O)), 2.24 (2H, m, CH CH = CFR), 1.23 (9H, s, C (CH) 3).
RMN '3C (7S MHz, CDC13), 8 (pprn): 200.94, 167.35, 160.39, 159.29, 157.13, 140.38, 130.54, 127.90, 127.43, 123.48, 115.16, 112.11, 105.48, 105.19, 65.09, 55.22, 52.33, 48.89, 39.05, 38.79, 33.53, 27.21, 26.84, 26.72, 21.96, 21.59 (4 more peaks).
1R (CHCl3), v (crri t): 3020, 2957, 2873, 1744, 1719, 1606, 1495, 1438, 1282, 1216, I 158, 1090, 1040, 695, 853, 756.

lss MS (m / e): 448 M +.
SMHR: calculated for CZSH3s06F (M ~: 448.2261, found: 448.2270 t 0.001.
Macrocycle Z2 EO
OPiv ------_ I / F
F ~
Me0 [~ OMe Me0, 18 O Ö 22 Solution 1: Ä a solution of pivalo ester 18 (1.67 g, 3.73 mmol) in the tetrahydrofuran (15 mL) degassed with argon was added N O-bis (trimethylsilyl) acetamide (1.8 mL, 7.46 mmol) and the mixture was heated at reflux for 5 h.
Solution 2: Ä a solution of tetrakis (triphenylphosphine) palladium (0) (1.07 g, 0.93 mmol) 1,3- was added to tetrahydrofuran (1.9 L) degassed with argon bis (diphenylphospino) propane (384 mg, 0.93 mmol) and the mixture was heated to reflux for 1 hour.
Solution 1 was added to solution 2 over a period of 14 h and the resulting mixture was heated at reflux for an additional 4 hours and then cooled. The solvent was evaporated under reduced pressure and the residue obtained was purified by flash chromatography (5% acetate ethyl, 35%
dichloromethane, 60% hexane) to give the macrocycle 22 in the form of a solid white (1.14g, 88%).
Gross formula: C2oH230aF.
Mp 129-131 ° C.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.21 (1H, d, J = 8.4 Hz, Ph), 6.73 (1H, dd, J = 8.4 Hz, and J = 2.8 Hz, Ph), 6.67 (1H, d, J = 2.8 Hz, Ph), 6.57 (1H, d, J = 15.7 Hz, PhCH = CH), 5.77 (IH, ddd, J = 15.6 Hz, J = 8.6 Hz and J = S. 3 Hz, CH = CHPh), 5.00 (1 H, dt, J = 20.7 Hz and J =
7.7 Hz, CH = CFR), 3.80 (3H, s, ArOCH), 3.77 (3H, s, C02CH), 3.80-3.75 (1H, m, C (O) CHC (O)), 3.04-1.86 (10H, m, PhCH CH CH = CFR, CH = C (F) CH CH C (O), CH C (I ~ C02Me).
13C NMR (75 MHz, CDCI3), 8 (ppm): 204.00, 169.54, 160.98, 159.09, 157.11, 140.77, 131.98, 129.24, 128.38, 126.07, 114.54, 111.63, 105.80, 105.52, 57.71, 55.19, 52.58, 39.06, 33.90, 32.01, 27.89, 27.78, 22.42, 22.03 (4 more peaks).
IR (CHC13), v (crri l): 3020, 2956, 1743, 1714, 1607, 1493, 1436, 1250, 1216, 1165, 1115, 1078, 1041, 970, 762.
MS (m / e): 346 M +.
SMHR: calculated for C2oH23O4F (M ~: 346.1580, found: 346.1576 ~ 0.0010.
Triene 15 EOEO
\ I ~ F ~ \ I ~ F
~ /
Me0 Me0 Ä a solution of benzeneseleninic anhydride 70% (10.2 g, 19.8 mmol) in the dichloromethane (200 mL) was added triethylamine (4.6 mL, 32.9 mmol) and the mixture has been stirred for 15 min. Ä this solution was added the macrocycle 22 by cannula and the mixture was stirred for 15 min. An aqueous solution of carbonate saturated sodium a was added and the aqueous phase was extracted with dichloromethane. The organic phases combined were dried with magnesium sulfate, filtered and concentrated.
The residue obtained was purified by flash chromatography (5% ethyl acetate, 95%
dichloromethane) to give triene 15 as a yellow solid (2.05 g, 91%).
Raw formula: CZOH21O4F.
Mp 147-149 ° C.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.43 (1H, d, J = 11.5 Hz, PhCH = CH = CI- ~ I, 7.21 (1H, d, J = 8.5 Hz, Ph), 6.96 (1 H, d, J = 16.2 Hz, CH = CHPh), 6.76 (1 H, dd, J =
8.4 Hz, and J = 2.7 Hz, Ph), 6.71 (1 H, d, J = 2.7 Hz, Ph), 6.64 (1 H, dd, J = 16.2 Hz and J =
11. 5 Hz, PhCH = CHI, 5.26 (1H, dt, J = 22.1 Hz and J = 8.7 Hz, CH = CFR), 3.82 (3H, s, ArOCH), 3.81 (3H, s, COZCH), 2.97-2.93 (2H, m, CH2CH C (O)), 2.87-2.81 (2H, m, PhCH CH2), 2.58 (2H, dt, J =
22.1 Hz and J = 6.9 Hz, CH CH2C (O)), 2.22-2.14 (2H, m, PhCH2CH).
13C NMR (75 MHz, CDC13), 8 (ppm): 203.17, 164.94, 160.41, 159.01, 155.73, 144.16, 144.00, 142.15, 136.13, 131.88, 126.46, 123.62, 117.45, 111.76, 107.23, 106.93, 55.29, 52.14, 39.48, 37.15, 26.14, 26.02, 23.61, 23.22 (4 more peaks).
IR (CHC13), v (crri l): 3020, 2954, 1696, 1603, 1583, 1436, 1275, 1241, 1158, 1077, 1045.
MS (m / e): 344 M +.
SMHR: calculated for C2oH2i0aF (M ~: 344.1424, found: 344.1426 ~ 0.0010.
Tetracycle 16 EOEO
F -Fi F
me0 me0 Ä a solution of triene 15 (500 mg, 1.45 mmol) in toluene (70 mL) degassed with argon a triethylamine (1.0 mL, 7.25 mmol) was added and the mixture was heated in a tube sealed at 165 ° C for 4 h. The mixture was cooled and the solvent was been evaporated under pressure scaled down. The residue obtained was purified by flash chromatography (20%
ethyl acetate, 80%
hexane) to give tetracycle 16 as a white solid (389 mg, 78%).
Gross formula: CZOH2 ~ 0aF.
Mp: 162-164 ° C.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.28 (1H, d, J = 8.9 Hz, Ph), 6.76 (1H, dd, J = 8.9 Hz and J = 2.7 Hz, ~, 6.70 (1 H, d, J = 2.7 Hz, ~, 6.53 (1 H, d, J = 9.9 Hz, CH = CHC (C02Me) R), 5.64 (1H, ddd, J = 9.9 Hz, J = 5.6 Hz and J = 3.0 Hz, CH = CHC (C02Me) R), 3.79 (3H, s, ArOCH), 3.72 (3H, s, C02CH), 3.23 (1H, d, J =
11.5 Hz, PhCHCH = CH), 3.00-2.94 (2H, m, PhCH CHz), 2.91-2.77 (1H, m, CHC (F) R), 2.51-2.23 (4H, m, C (O) CH CH C (F) R), 2.14-1.92 (1H, m, PhCH2CHI-- ~ I, 1.73-1.57 (1H, m, PhCH2CIrII ~.
'3C NMR (75 MHz, CDC13), 8 (ppm): 209.92, 167.91, 157.95, 137.43, 132.10, 128.26, 125.88, 122.52, 114.30, 111.89, 105.07, 102.64, 65.61, 55.25, 52.88, 40.66, 40.37, 40.13, 40.01, 35.60, 29.51, 26.43, 26.10, 21.36 (4 more peaks).
IR (CHCl3), v (crri '): 3019, 2954, 1761, 1734, 1609, 1502, 1436, 1258, 1216, 1045.
MS (m / e): 344 M +.
SMHR: calculated for C2oH21O4F (M ~: 344.1424, found: 344.1413 ~ 0.0010.
Alcohol 59 EOE OH
I \ F - ~ ¿\ F
/ ~ \ /
Me0 ~ Me0 to a solution of ketone 15 (205 mg, 0.60 mmol) in a mixture of methanol and of 1: 1 dichloromethane (10 mL) was added cerium chloride heptahydrate (223 mg, 0.60 mmol) and sodium borohydride (25 mg, 0.60 mmol). The mixture resulting was stirred at room temperature for 5 min and an aqueous solution of chloride saturated ammonium was added. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (30%
ethyl acetate, 70% hexane) to give alcohol 59 in the form of a beige solid (182 mg, 88%).
Raw formula: C2oHZ3O4F.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.45 (1H, d, J = 9.9 Hz, PhCH = CH = C) ~, 7.22 (1H, d, J = 8.4 Hz, Ph), 6.93-6.72 (4H, rn, Ph, PhCH = CH, ~, 5.27 (1H, ddd, J = 23.2 Hz, J = 13.2 Hz and J = 4.3 Hz, CH = CFR), 4.72 (1 H, t, J = 10.5 Hz, RR 'CHOH), 4.12 (1 H, d, J = 10.5 Hz, RR'CHOI- ~ I, 3.821 (3H, s, ArOCI- ~ I), 3.815 (3H, s, COZCH), 3.04 (1H, td, J
= 12.9 Hz and J =
2.0 Hz, PhCHHCH2), 2.80 (1 H, td, J = 12.9 Hz and J = 6.3 Hz, PhCHHCH2), 2.42-1.96 (6H, m).
13C (75 MHz, CDCl3), 8 (ppm): 167.13, 160.06, 159.54, 155.87, 142.14, 141.94, 141.89, 136.31, 129.07, 123.38, 117.62, 111.69, 106.90, 106.58, 67.32, 55.29, 51.82, 38.10, 38.07, 32.71, 24.70, 24.58, 23.69, 23.32 (5 more peaks).
IR (CHCI3), v (crri '): 3519, 3019, 2955, 1681, 1601, 1561, 1438, 1270, 1216, 1160, 758.
MS (m / e): 346 M +.
SMHR: calculated for C2oH23O4F (M ~: 346.1580, found: 346.1587 ~ 0.0010.
Silyl ether 60a OTBDMS
~ F
W / W /
Me0 ~ Me0 59 60a Ä a solution of alcohol 59 (180 mg, 0.52 mmol) in tetrahydrofuran (5 mL) have been added imidazole (156 mg, 2.6 mmol) and t-butyldimethylsilane chloride (392 mg, 2.6 mmol) and the mixture was stirred at room temperature overnight. A
aqueous solution saturated ammonium chloride was added and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried with sulfate of magnesium, filtered and concentrated under reduced pressure. The residue obtained has been purified by flash chromatography (10% ethyl acetate, 90% hexane) to give the ether silylated 60a as a pale yellow (217 mg, 92%).
Raw formula: C26H3 ~ O4FS1.
1 H NMR (300 MHz, CDCI3), 8 (ppm); 7.54-7.34 (2H, m, PhCH = CH = Ces, 7.22 (1H, d, J =
8.3 Hz, Ph), 6.81-6.72 (3H, m, Ph, PhCH = CH = CH), 5.16-5.03 (2H, m, CH = CFR, RR'CHOTBDMS), 3.82 (3H, s, ArOCH), 3.74 (3H, s, C02CH), 3.06 (1H, td, J =
13.0 Hz and J = 3.3 Hz, PhCHHCH2), 2.78 (1H, td, J = 13.0 Hz and J = 5.4 Hz, PhCHHCH2), 2.61-1.94 (5H, m), 1.82-1.74 (1H, m), 0.84 (9H, s, Si (CH3) 3), 0.06 (3H, s, SiCH3), 0.04 (3H, s, SiCH3).
13C NMR (75 MHz, CDC13), 8 (ppm): 167.89, 160.14, 159.68, 157.31, 142.61, 142.02, 140.76, 136.07, 131.04, 129.08, 125.85, 117.36, 111.50, 104.07, 103.75, 70.34, 55.25, 51.71, 37.67, 21.14, 25.78, 25.24, 25.11, 22.52, 22.15, -5.31, -5.43 (4 peaks more).
IR (CHCl3), v (cm 1): 3020, 2954, 2857, 1695, 1597, 1436, 1271, 1215, 1160, 1044, 928, 837.
5M (m / e): 403 (M-C4H9) +.
SMHR: calculated for C22H2804FSi (M ~: 403.1741, found: 403.1734 ~ 0.0012.
Tetracycles 63a and 64b E OTBDMS E OTBDMS E OTBDMS
F
HF + ~ ~ HF
Me0 Me0 Me0 60a 63a 64a Ä a solution of triene 60a (224 mg, 0.49 mmol) in toluene (70 mL) degassed with argon triethylamine (345 p, L, 2.45 mmol) was added and the mixture was heated in a tube sealed at 220 ° C for 16 h. The mixture was cooled and the solvent was been evaporated under reduced pressure. The residue obtained was purified by flash chromatography (5% acetate ethyl, 95% hexane) to give separable tetracycles 63a and 64a and unstable under as a white solid (142 mg, 63%).
Alcoo1, Q63a (majority) Gross formula: C26H37 ~ 4FS1.
5M (m / e): 403 (M-C4I-i9) +.

161.
1H NMR (300 MHz, CDCl3), 8 (ppm): 7.31 (1H, d, J = 8.6 Hz, ~, 6.76 (1H, dd, J
= 8.6 Hz and J = 2.7 Hz, Ph), 6.6 8 (1 H, d, J = 2.7 Hz, P ~, 6.49 (1 H, d, J = 10.2 H
z, CH = CHC (C02Me) R), 5.76 (1H, ddd, J = 10.2 Hz, J = 5.3 Hz and J = 3.0 Hz, CH = CHC (COZMe) R), 5.04 (1H, dd, J = 9.8 Hz and J = 6.4 Hz, CHOTBDMS), 3.79 (3H, s ArOCH), 3.68 (3H, s, COzCH), 3.19 (1H, d, J = 11.1 Hz, PhCHCH = CH), 2.94-2.89 (2H, m, PhCH CH2), 2.45-2.34 (1H, m, CHC (F) R), 2.27-1.40 (6H, m), 0.88 (9H, s, SiC (CH
) 3), 0.11 (6H, d, J = 2.3 Hz, Si (CH) 2).
13C NMR (75 MHz, CDCl3), 8 (ppm): 157.73, 137.76, 131.10, 126.19, 114.08, 111.83, 111.50, 105.81, 103.12, 74.97, 55.23, 52.21, 41.45, 41.14, 39.36, 39.24, 31.06, 29.65, 29.02, 28.68, 25.83, 25.72, 21.17, 21.14, -4.63, -4.98 (3 more peaks).
IR (CHCl3), v (crri '): 3019, 2954, 2930, 2857, 1730, 1502, 1255, 1215, 1140, 758.
Alcohol 63a-1 OTBDMS E OH
/ /
HF ~ ~ \ Fi F
Me0 I / Me0 /
63a 63a-1 Ä a solution of silyl ether 63a (142 mg, 0.31 mmol) in methanol (10 mL) was added p-toluenesulfonic acid and the mixture was stirred at temperature ambient all the night. A saturated aqueous sodium bicarbonate solution was added and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried in magnesium sulfate, filtered and evaporated. The residue obtained has been purified through flash chromatography (30% ethyl acetate, 70% hexane) to give alcohol 63a-1 sub form of a colorless oil (80 mg, 75%).

Raw formula: CZOH2304F.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.30 (1H, d, J = 8.5 Hz, Phi, 6.77 (1H, dd, J = 8.5 Hz etJ = 2.7Hz, Phi, 6.70 (lH, d, J = 2.7Hz, P ~, 6.57 (lH, d, J = 10.1 Hz, CH = CHC (C02Me) R), 5.77 (1H, ddd, J = 10.1 Hz, J = 5.7 Hz and J = 3.1 Hz, CH = CHC (COZMe) R), 4.9I (1H, dd, J = 11.0 Hz and J = 6.6 Hz, CHOOSE, 3.79 (3H, s, ArOCH), 3.71 (3H, s, C02CH), 3.22 (IH, d, J = I 1.2 Hz, PhCHCH = CH), 2.97-2.92 (2H, m, PhC ~ CH2), 2.63-2.55 (1H, m, CHC (F) R), 2.22-1.48 (7H, m).
13C NMR (75 MHz, CDCl3), 8 (ppm): 172.20, 172.14, 157.82, 137.77, 134.66, 132.77, 129.20, 125.89, 122.40, 122.33, 114.20, 111.77, 105.64, 103.21, 74.34, 55.25, 52.50, 41.02, 40.71, 39.79, 39.67, 29.39, 28.53, 28.44, 28.18, 21.11, 21.07 (7 peaks more).
IR (CHCl3), v (crri l): 3588, 3019, 2954, 1722, 1609, 1502, 1435, 1259, 1215, 1045, 726.
5M (m / e): 346 M +.
SMHR: calculated for C2oH23O4F (M ~: 346.1580, found: 346.1569 t 0.0010.
Tetracycle 72 EOEO
i -/ Fi F ~ ~ HF
Me0 Me0 vv Ä a solution of tetracycle 16 (594 mg, 1.73 mmol) in absolute ethanol (75 mL) was added 5% palladium (0) on barium sulfate (745 mg, 0.35 mmol) and hydrogen has been bubbled for 15 min. The resulting mixture was stirred under an atmosphere of hydrogen all the overnight at room temperature. The solution was filtered through silica and the solvent has been evaporated to give tetracycle 72 as a beige solid (594 mg, 99%).

Gross formula: C2pH23 ~ 4F ~
Mp: 154-157 ° C.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.19 (1H, d, J = 8.6 Hz, Ph), 6.74 (1H, dd, J = 8.6 Hz and J = 2.7 Hz, Ph), 6.65 (1H, d, J = 2.7 Hz, ~, 3.78 (3H, s, ArOCH), 3.73 (3H, s, C02CH), 2.94-2.89 (2H, m, PhCH CH2), 2.83-2.70 (1H, m, PhC ~, 2.58-2.49 (2H, m, RC (O) CH
) 2.38-1.90 (7H, m), 1.65-1.77 (2H, m).
13C NMR (75 MHz, CDC13), 8 (ppm): 211.61, 169.27, 169.23, 157.77, 137.43, 130.21, 127.05, 113.72, 112.08, 106.08, 103.66, 62.19, 61.94, 55.16, 52.30, 42.70, 4240, 39.37, 39.24, 34.97, 30.12, 30.05, 29.79, 26.73, 26.19, 25.86, 22.04 (6 peaks more).
IR (CHC13), v (crri l): 3055, 2954, 1754, 1731, 161 l, 1502, 1435, 1266, 1043, 896.
MS (m / e): 346 M +.
SMHR: calculated for C2oH23O4F (M ~: 346.1580, found: 346.1576 f 0.0010.
Triflic enol ether 79 OE OTf / HF ~~ (/ Fi F
Me0 Me0 72 7g Ä a solution of dopropopropylethylamine (290 p, 1 ,, 2.20 mmol) in the tetrahydrofuran (15 mL) at 0 ° C was added a solution of 1.6 M n-butyl lithium in hexane (1.3 mL, 2.13 mrnol) and the mixture was stirred at 0 ° C for 20 min. The solution has been cooled to -78 ° C and ketone 72 (245 mg, 0.71 mmol) was added by cannula. The mixture resulting was stirred at -78 ° C for 15 min then at 0 ° C for an additional 15 min.
N (5-chloro-2-pyridyl) triflimide (507 rng, 1.42 mmol) in tetrahydrofuran (5 mL) was added by cannula at 0 ° C and the mixture was stirred at room temperature during I h, water was added and the aqueous phase was extracted with ethyl acetate. The organic phases combined were dried with magnesium sulfate, filtered and evaporated.
The residue obtained was purified by flash chromatography (20% ethyl acetate, 80% hexane) to give enol triflic ether 79 as a yellow oil (306 mg, 90%
Raw formula: C21Ha2O6FaS.
1H NMR (300 MHz, CDCI3), 8 (ppm): 7.22 (1H, d, J = 8.6 Hz,> ~, 6.75 (1H, dd, J = 8.6 Hz and J = 2.8 Hz, ~, 6.65 (1 H, d, J = 2.8 Hz, ~, 5.75 (1 H, t, J = 2.5 Hz, CH = C (OTf) R), 3.79 (3H, s, ArOCH), 3.76 (3H, s, C02CH), 2.98-2.57 (5H, m), 2.47-2.39 (1H, m), 2.32-2.13 (3H, m), 1.96 (1H, qd, J = 13.1 Hz and J = 3.4 Hz), 1.64-1.39 (2H, m).
13C NMR (75 MHz, CDCl3), b (ppm): 169.59, 169.50, 157.77, 149.96, 137.53, 129.74, 127.64, 120.56, 116.32, 113.69, 112.94, 112.28, 111.80, 104.55, 102.08, 59.59, 59.32, 55.18, 52.20, 43.11, 42.80, 39.20, 39.07, 34.31, 33.94, 33.83, 33.77, 29.68, 26.45, 21.93 (9 peaks of more).
IR (CHCI3), v (crri '): 2952, 2840, 1710, 1655, 1611, 1502, 1424, 1272, 1217, 1142, 1044, 968, 887.
5M (m / e): 478 M +.
SMHR: calculated for C2] H2zO6F4S (M '~: 478.1073, found: 478.1060 ~ 0.0014.
Nitrite c ~~ unsaturated 80 OTf E CN
\. 1 "\
hi FI Fi F
Me0 ~ Me0 Ä a solution of the triflic enol ether 79 (96 mg, 0.20 mmol) in the benzene (10 mL) have lithium cyanide (40 mg, 1.20 mmol), tétralcis (triphenylphosphine) palladium (0) (23 mg, 0.02 mmol) and crown ether 12-C-4 (3 ~ L, 0.02 mmol).
The mixture resulting was stirred at room temperature overnight and catalyst palladium (23 mg, 0.02 mmol) has been added. The mixture was stirred for 24 h at temperature room then water has been added. The aqueous phase was extracted with acetate ethyl and phases combined organic were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (20% ethyl acetate, 80% hexane) to give nitrite a, ~ 3-unsaturated 80 as a solid (49 mg, 69%).
Raw formula: C21Hz20sFN.
1H NMR (300 MHz, CDC13), 8 (ppm): 7.22 (1H, d, J = 8.6 Hz, P_h ,, 6.75 (1H, dd, J = 8.6 Hz and J = 2.7 Hz, Ph), 6.69 (1 H, t, J = 2.5 Hz, CH = C (CN) R), 6.66 (1 H, d, J =
2.7 Hz, Ph), 3.792 (3H, s, ArOCH), 3.789 (3H, s, C02CH), 2.99-2.82 (3H, m), 2.75 (IH, d, J =
2.5 Hz, CH = C (C1 ~ CHI- ~ I, 2.61 (1H, dq, J = 14.4 Hz and J = 3.2 Hz), 2.46-2.17 (4H, m), 1.94 (1H, qd, J = 13.5 Hz and J = 3.6 Hz), 1.52-1.35 (2H, m).
RMN I3C (75 MHz, CDCl3), 8 (ppm): 169.95, 169.86, 157.78, 144.03, 137.52, 129.59, 129.55, 127.55, 127.32, 123.53, 120.44, 115.23, 113.75, 112.26, 106.34, 103.90, 62.59, 62.31, 55.23, 52.47, 42.62, 42.32, 39.39, 29.26, 38.96, 38.60, 34.88, 34.81, 29.76, 26.20, 22.79 (10 more peaks).
IR (CHCl3), v (crri l): 3055, 2955, 2228, 1737, 161 l, 1502, 1422, 1262, 1212, 1042, 892.
MS (m / e): 355 M +.
SMHR: calculated for C2IHZZO3FN (M ~: 355.1584, found: 355.1592 t 0.001 I.
Diols 73 > H
-., \ Fi F
me0 Ä a solution of ester 72 (575 mg, 1.66 mmol) in tetrahydrofuran (40 mL) have been added lithium borohydride (180 mg, 8.30 mmol) and methanol (336 p, L, 8.30 mmol) and the mixture was stirred at room temperature for 2 h. Of lithium borohydride (72 mg, 3.32 mmol) and methanol (134 p, L, 3.32 mmol) were added and the mixed The resulting was stirred at room temperature for an additional 1 hr. A
solution saturated ammonium chloride was added and the aqueous phase was been extracted with ethyl acetate. The combined organic phases were dried with sulfate magnesium, filtered and concentrated. The residue obtained was purified by flash chromatography (100% ethyl acetate) to give a separable mixture 9: I (a:, l ~ de diol 73 in the form of a white solid (539 mg, 94% overall).
Raw formula: Cl9HasO3F.
3 C NMR (75 MHz, CDCI3), 8 (ppm): Insoluble and decomposes in solution.
1R (CHC13), v (cm '): Insoluble.
MS (m / e): 320 Mt.
SMHR: calculated for Cl9Hzs03F (M ~: 320.1788, found: 320.1779 ~ 0.0010.
Majority Diol a ) ZMN'H (300 MHz, CDC13), 8 (ppm): 7.22 (1H, d, J = 8.6 Hz, P. ~ h, 6.72 (1H, dd, J = 8.6 Hz and J = 2.7 Hz, Ph), 6.64 (1 H, d, J = 2.7 Hz, Ph), 4.74 (1 H, t, J = 8.2 Hz, RR 'CHOH), 4.00 (IH, d, J = 10.3 Hz, CHHOH), 3.90 (1H, d, J = 10.3 Hz, CHHOH), 3.78 (3H, s, ArOCH), 2.85 (2H, dd, J = 8.9 Hz and J = 3.8 Hz, PhCH CHZ), 2.47 (1H, t, J = 11.6 Hz, PhCH ~, 2.38-2.05 (5H, m), 1.92-1.62 (3H, m), 1.87 (2H, solid, CHOH and CH20 ~, 1.50-1.28 (3H, m).
Minority Diol / 3 1 H NMR (300 MHz, CDCI3), S (ppm): 7.18 (1H, d, J = 8.6 Hz, ~, 6.73 (1H, dd, J
= 8.6 Hz and J = 2.7 Hz, Ph ,, 6.64 (1H, d, J = 2.7 Hz, ~, 4.18 (1H, d, J = 11.7 Hz, RCHHOH), 4.01 (1 H, dd, J = 7.3 Hz and J = 2.2 Hz, RR 'CHOH), 3.78 (3H, s, ArOCH), 3.71 ( 1 H, d, J = 11.7 Hz, CHHOH), 2.86 (2H, dd, J = 8.6 Hz and J = 3.5 Hz, PhCH CHZ), 2.49-2.35 (2H, m), 2.31-1.83 (10H, m), I.74 (1H, qd, J = I 1.7 Hz and J = 2.0 Hz), 1.53-1.34 (1H, m), I.OS-0.96 (1H, m).

Tosylates 85 ) H) H

Ä a solution of the diols 73 (215 mg, 0.67 mmol) in dichloromethane (50 mL) have been added triethylamine (281 p, L, 2.01 mmol), p- chloride toluenesulfonyl (193 mg, 1.01 mmol) and 4-dimethylaminopyridine (8 mg, 0.067 mmol). The mixture resulting has been stirred at room temperature overnight and an aqueous solution chloride saturated ammonium was added. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (50%
ethyl acetate, 50% hexane) to give the mixture of tosylates 85 as a solid white in a 9: 1 mixture (a: (3) non-separable (277 mg, 87%).
OR
Ä a solution of the diols 73 (460 mg, 1.4 mmol) in a mixture of benzene and of dimethylformamide 3: 1 (40 mL) was added dibutylstannane oxide (430 mg, 1.73 mmol) and the mixture was brought to reflux with a dean-stark for 1.5 h. The solution was cooled to room temperature and p-toluenesulfonyl chloride was added. The mixture was stirred at room temperature for 1 h and a solution aqueous chloride of saturated ammonium was added. The aqueous phase was extracted with a mixed 1: 1 hexane and ether and the combined organic phases were dried with sulfate magnesium, filtered and concentrated. The residue obtained was purified by flash chromatography (50% ethyl acetate, 50% hexane) to give the mixture of tosylates 85 under shape of a white solid in a non-separable 9: 1 (a: Vii) mixture (583 mg, 87%).

Tosylate a 85 (majority) Raw formula: C26HstOsFS.
1 H NMR (300 MHz, CDCI3), b (ppm): 7.78 (2H, d, J = 8.2 Hz, Ph), 7.34 (2H, d, J
= 8.2 Hz, Ph), 7.16 (1 H, d, J = 8.6 Hz, ~, 6.72 (1 H, dd, J = 8 .6 Hz and J = 2.5 Hz, Ph), 6.62 (IH, d, J =
2.5 Hz, Ph, 4.81 (1H, solid, RR'CHOI- ~ i, 4.57 (1H, t, J = 8.0 Hz, RR'CHOH), 4.30 (IH, d, J = 9.9 Hz, CHHOTs), 4.23 (1H, d, J = 9.9 Hz, CHHOTs), 3.78 (3H, s, ArOCH), 2.86-2.79 (2H, m, PhCH CH2), 2.44 (3H, s, PhCH ~, 2.37-2.1 S (2H, m), 2.08-2.01 (3H, m), 1.88-1.52 (4H, m), I.46-1.33 (2H, m), 1.29-1.09 (1H, m).
13C NMR (75 MHz, CDCl3), 8 (ppm): Decomposes in solution.
IR (CHC13), v (crri l): 3602, 3SSI, 2951, 2872, 1610, ISO1, 1464, 1361, 1176, 1097, 1045, 944, 816.
MS (male): 474 M +.
SMHR: calculated for C26HsiOsFS (M ~: 474.1876, found: 474.1880 t 0.0014.
Alcohol 87 and ozirane 86 OTs ~ OH MQ UH O
~ hi F ~ ~ HF + ~ r hi F
Me0 Me0 Me0 Ä a mixture solution of tosylates 85 (6S0 mg, 1.37 mmol) in Ie dimethylsulfoxide (80 mL) was added sodium borohydride (260 mg, 6.85 mmol) and mixture was heated to 80 ° C overnight. An aqueous chloride solution saturated ammonium a was added and the aqueous phase was extracted with a mixture of ether and of hexane I: I. The the combined organic phases were dried with magnesium sulfate, filtered and concentrated under reduced pressure. The residue obtained was purified by flash chromatography (50% ethyl acetate, SO% hexane) to give alcohol 87 in the form of a colorless oil (353 mg, 85%) and approximately 10% of oxirane 86.

Alcohol 87 Gross formula: C19H25 ~ 2F ~
1H NMR (300 MHz, CDCl3), 8 (ppm): 7.22 (1H, d, J = 8.6 Hz, Ph), 6.73 (IH, dd, J = 8.6 Hz and J = 2.8 Hz, Phi, 6.65 (1H, d, J = 2.8 Hz, Pha, 4.28 (1H, t, J = 8.0 Hz, RR'CHOH), 3.78 (3H, s, ArOCH), 2.90-2.85 (2H, m, PhCH CH2), 2.46-2.39 (1H, m, PhCI- ~, 2.32-2.00 (4H, m), 1.90-1.74 (2H, m), 1.72-1.57 (3H, m), 1.53-1.36 (3H, m), 1.08 (3H, d, J =
1.3 Hz, CH).
13C NMR (75 MHz, CDCl3), b (ppm): 157.58, 137.76, 131.39, 126.83, l I3.71, 111.88, 109.08, 106.71, 80.43, 55.21, 47.48, 47.25, 42.40, 42.11, 40.36, 40.23, 29.97, 28.42, 28.33, 28.22, 27.58, 27.24, 25.68, 22.13, 16.33, 16.25 (7 more peaks).
IR (CHCl3), v (cm ''): 3386, 2942, 2870, 1610, 1501, 1466, 1261, 1235, 1042.
MS (m / e): 304 M +.
SMHR: calculated for C19H2502F (M ~: 304.1838, found: 304.1843 ~ 0.0009.
Oxirane 86 Raw formula: CIgH23O2F.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 7.12 (1 H, d, J = 8.6 Hz, ~, 6.72 (1 H, dd, J = 8.6 Hz and J = 2.8 Hz, l ~, 6.62 (1 H, d, J = 2.8 Hz, ~, 4.95 (IH, d, J = 5.9 Hz, CHHOCHRR '), 4.72 (IH, d, J ~ 5.9 Hz, CHHOCHRR '), 4.35 (1H, dd, J = 5.6 Hz and J = 4.7 Hz, RR'CHOCH2) 3.78 (3H, s, ArOCH), 2.90-2.84 (2H, rn, PhCH CH2), 2.48-1.25 (12H, m).
13C NMR (75 MHz, CDCl3), 8 (ppm): 157.51, 137.58, 131.72, 127.84, 113.38, 112.33, I06.60, 104.18, 91.12, 91.07, 74.36, 74.16, 55.20, 51.18, 50.95, 44.32, 43.97, 36.66, 36.51, 32.33, 32.16, 32.12, 31.98, 30.31, 29.05, 27.83, 23.94 (8 more peaks).
TR (CHCl3), v (cry 1): 3018, 2945, 2882, 1610, 1502, 1465, 121 S, 1038, 974.
MS (m / e): 302 M +.
SMHR: calculated for C19H2302F (M ~: 302.1682, found: 302.1399 ~ 0.0009.

Ketone 14 Me ~ H Me ~
HF ~ \ HF
Me0 ~ Me0 ~
87 '14 Ä a solution of alcohol 87 (312 mg, 1.03 mmol) in dichloromethane (50 mL) have been added the molecular sieve 4 ~ (515 mg), the N oxide of 4-methylmorpholine (241 mg, 2.06 mmol) and tretrapropylammonium perruthenate (35 mmg, O.IO mmoI). The mixed resulting was stirred at room temperature for 2 h and then filtered through silica. The filter was evaporated and the residue obtained was purified by flash chromatography (30% acetate ethyl, 70% hexane) to give the ial ketone in the form of a white soda (269 mg, 87ç ~ ô).
Gross formula: C19H2302F ~
1H NMR (300 MHz, CDC13), 8 (ppm): 7.19 (1H, d, J = 8.6 Hz, ~, 6.74 (1H, dd, J
= 8.6 Hz and J = 2.8 Hz, ~, 6.67 (1H, d, J = 2.8 Hz, PhJ, 3.79 (3H, s, ArOCH), 2.93 (2H, dd, J = 9.0 Hz and J = 3.9 Hz, PhCH CH2), 2.59-2.48 (3H, m, PhCH and RC (O) CH), 2.42-2.13 (4H, m), 1.92 (1H, qd, J = 11.9 Hz and J = 2.1 Hz), 1.65-I.4I (4H, m), 1.13 (3H, d, J =
1.5 Hz, CH).
13C NMR (75 MHz, CDCI3), 8 (ppm): 219.62, 157.80, 137.56, 130.58, 126.78, 113.82, 112.03, 107.01, 104.64, 55.21, 52.94, 52.67, 42.19, 41.90, 40.27, 40.14, 33:30, 32.42, 32.34, 29.90, 25.57, 25.21, 21.97, 12.82, 12.72 (6 more peaks).
IR (CHCl3), v (crri l): 2944, 2869, 1741, 161 l, 1502, 1464, 1274, 1232, 1093, 1043, 936.
MS (m / e): 302 M +.
SMHR: calculated for C29H23OZF (M ~: 302.1682, found: 302.1689 ~ 0.0009.

Tritlic Enol Ether 88 Ms ~ Me ~ Tf ~ HF ~ ~ / HF
Me0 Me0 Ä a solution of ketone 14 (22 mg, 0.073 mmol) in tetrahydrofuran (2 mL) at 0 ° C a was added a solution of bis (trimethylsilyl) potassium amide 0.5 M
in toluene (740 ~ L, 0.37 mmol) and the mixture was stirred at 0 ° C 15 min and then at ambient temperature for 4S min. N (5-chloro-2-pyridyl) triflimide (93 mg, 0.22 mmol) in the tetrahydrofuran (1 mL) was added by cannula and the mixture was stirred at temperature room for 30 min. an aqueous solution of saturated ammonium chloride has been added and the aqueous phase was extracted with ethyl acetate. 'The organic phases combined were dried with magnesium sulfate, filtered and evaporated.
The residue obtained was purified by flash chromatography (5% ethyl acetate, 95% hexane) to give enol triflic ether 88 as a pale yellow oil (27 mg, 84%).
Gross formula: C2pH ~ O4F4S.
1 H NMR (300 MHz, CDCI3), 8 (ppm): 7.22 (IH, d, J = 8.6 Hz, ~, 6.74 (1H, dd, J
= 8.6 Hz and J = 2.7 Hz, Ph>, 6.65 (IH, d, J = 2.7 Hz, Phi, 5.59 (1H, t, J = 2.7 Hz, CH = C (OTfjR), 3.79 (3H, s, ArOCH), 2.93-2.86 (2H, m, PhCH CH2), 2.71 (1H, td, J = 17.1 Hz and J
= 1.8 Hz, PhCH ~, 2.61-2.40 (2H, m), 2.30-2.05 (3H, m), 1.87 (IH, qd, J = 12.0 Hz and J =
2.1 Hz), 1.57-1.26 (3H, m), I .I 8 (3H, d, J = 2.7 Hz, CH).
1Z1VIN 13C (75 MHz, CDCI3), 8 (pprn): 157.77, 155.04, 137.57, 130.16, 127.23, 120.65, 116.40, 113.76, 112.12, 108.48, 105.33, 102.86, 55.21, 49.48, 49.22, 43.00, 42.70, 40.39, 40.25, 37.22, 37.15, 34.05, 33.68, 29.57, 25.54, 22.27, 13.31, 13.16 (8 peaks Moreover).
IR (CHC13), v (cni l): 2944, 2868, 1650, 161 I, 1502, 1423, 1218, 1141, 1085, 1044, 884.
MS (m / e): 434 M +.

SMHR: calculated for C2pH22 ~ 4F4s (~: 434.1175, found: 434.1181 ~ 0.0013.
Nitrite c ~~ -unsaturated 89 Me ~ Tf My CN
\ \
HF t1 F
Me0 Me0 Ä a solution of triflic enol ether 88 (41 mg, 0.094 mmol) in benzene (10 mL) have lithium cyanide (18 mg, 0.56 mmol), tetrakis (triphenylphosphine) palladium (0) (11 mg, 0.0094 mmol) and crown ether 12-C-4 (2 ~ L, 0.0094 mmol). The resulting mixture was stirred at room temperature overnight and catalyst palladium (11 mg, 0.0094 mmol) has been added. The mixture was stirred 24 h at Ia temperature ambient then water was added. The aqueous phase was extracted with acetate ethyl and the combined organic phases were dried with sodium sulfate magnesium, filtered and evaporated. The residue obtained was purified by chromatography flash (10% acetate ethyl, 90% hexane) to give the nitrite a, ~ unsaturated 89 in the form of a solid (23 mg, 79%).
Gross formula: C2oH ~ OFN.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.22 (1 H, d, J = 8.6 Hz, Phi, 6.74 (1 H, dd, J = 8.6 Hz and J = 2.7 Hz, Phi, 6.65 (1H, d, J = 2.7 Hz, PhJ, 6.59 (1H, t, J = 2.6 Hz, CH = C (CN) R), 3.79 (3H, s, ArOCH), 2.93-2.88 (2H, m, PhCH CH2), 2.84-2.68 (IH, m), 2.47-2.38 (1H, m), 2.31-2.10 (3H, m), 1.90 (1H, qd, J = 11.9 Hz and J = 2.1 Hz), 1.57-1.22 (4H, m), 1.31 (3H, d, J
= 2.6 Hz, CH).
ItMN 13C (75 MHz, CDC13), 8 (ppm): 157.79, 142.09, 137.52, 127.22, 113.78, 112.12, 107.07, 104.62, 55.22, 52.57, 52.31, 42.78, 42.48, 40.35, 40.22, 38.99, 38.63, 37.93, 37.86, 29.94, 29.70, 25.69, 23.10, 14.96, 14.83 (S more peaks).

IR (CHC13), v (crri l): 3054, 2932, 2568, 2219, 1610, 1502, 1422, 1265, 1041, 883.
MS (m / e): 311 M +.
SMHR: calculated for C2oH22OFN (M ~: 311.1585, found: 3I I.I678 ~ 0.0009.
Nitriles 90a and 90b Me CN Me CN Me CN
'~ _ +
H Fv HF HF
Me0 Me0 Me0 89 90b 90a Ä a solution of nitrite 89 (20 mg, 0.064 mmol) in absolute ethanol (2 mL) was added on 5% palladium (0) on activated charcoal (28 mg, 0.013 mmol) and hydrogen has been hilled for 10 min. The mixture was stirred under a hydrogen atmosphere for 2 h then filtered on silica. The solvent was evaporated and the residue obtained was purified by chromatography flash (20% ethyl acetate, 80% hexane) to give a 1.6: 1 mixture ((3:
a) nitriles 90b and 90a separable in the form of a white solid (19 mg, 95%).
Gross formula: C2pH2qOFN.
MS (m / e): 313 M +.
SMHR: calculated for C2oH24C; FN (M '~: 313.1842, found: 313.1849 ~ 0.0009.
Nitrite ~ 90b (majority) 1 H NMR (300 MHz, CDCl3), 8 (ppm): 7.16 (1H, d, J = 8.6 Hz, PhJ, 6.74 (1H, dd, J = 8.6 Hz and J = 2.8 Hz, Pte, 6.65 (IH, d, J = 2.8 Hz, Phi, 3.78 (3H, s, ArOCH), 2.92-2.87 (2H, m, PhCH CHZ), 2.74 (1H, t, J = 7.7 Hz, R (CN) CHCH2), 2.50-2.42 (1H, m, PhC ~ - ~ I, 2.35-2.15 (5H, m), 2.10-2.03 (1H, m), 1.86 (IH, qd, J = 11.8 Hz and J = 2.2 Hz), 1.71-1.64 (1H, m), 1.57-1.36 (3H, m), 1.33 (3H, d, J = 1.1 Hz, CH).

'3C NMR (7S MHz, CDC13), S (ppm): 157.76, 137.61, 130.64, 126.70, 121.71, 113.73, 112.05, 109.19, 106.78, SS.21, 48.52, 48.26, 42.03, 41.75, 39.93, 39.80, 36.93, 36.85, 30.32, 30.02, 26.64, 25.72, 23.17, 16.38, 16.29 (S more peaks).
IR. (CHCI3), v (crri '): 2932, 2868, 2238, 1610, 1501, 1466, 1236, 1161, 1038.
X-ray: diffusion of hexane in ethyl acetate.
Nitrite at 90a (minority) 1 H NMR (300 MHz, CDC13), 8 (ppm): 7.20 (1H, d, J = 8.6 Hz, ~, 6.74 (1H, dd, J
= 8.6 Hz and J = 2.8 Hz,>'h, 6.65 (1H, d, J = 2.8 Hz, Phi, 3.78 (3H, s, ArOCH), 3.04 (1H, t, J = 9.2 Hz, R (CN) CHCH2), 2.91-2.86 (2H, m, PhCH CH2), 2.47 (1H, td, J = 11.2 Hz and J = 3.S
Hz, PhCH), 2.36-1.77 (8H, m), 1.53-1.33 (3H, m), 1.18 (3H, d, J = 1.0 Hz, CH).
'3C NMR (7S MHz, CDC13), 8 (ppm): 157.77, 137.47, 130.64, 126.78, 120.51, 113.78, 111.99, SS.21, 42.02, 41.73, 40.10, 39.97, 39.13, 31.64, 31.57, 29.90, 29.68, 29.35, 25.85, 24.31, 22.88, 16.81, 16.72 (3 more peaks).
IR (CHCl3), v (cm ''): 2938, 2239, 1610, 1502, 1467, 1320, 1236, 1043, 957.
Bromide 91 Me CN Me CN
"'-' ~ ~
Fi F ~ / H Br Me0 r TBDMSO
90b 91 Ä a solution of nitrite 90b (36 mg, 0.12 mmol) in dichloromethane (S
mL) at -30 ° C a was added a solution of boron tribromide 1 M in dichloromethane (1.8 mL, 1.80 mmol) and the mixture was stirred at 0 ° C for 3 h. Methanol was added followed by water and the aqueous phase was extracted with dichloromethane. Organic phases gathered have been dried with magnesium sulfate, filtered and evaporated. The residue obtained has been purified by flash chromatography (40% ethyl acetate, 60% hexane) to give the phenol corresponding in the form of a more or less pure oil (30 mg).
to a solution of this phenol (30 mg, 0.083 mmol) in dichloromethane (5 mL) have been added imidazole (1S mg, 0.25 mmol) and t-butyldimethylsilane chloride (23 mg, 0.15 mmol) and the mixture was stirred at room temperature for 3 h. A
aqueous solution saturated ammonium chloride was added and the aqueous phase was extracted with dichloromethane. The combined organic phases were dried with sulfate of magnesium, filtered and concentrated under reduced pressure. The residue obtained has been purified by flash chromatography (15% ethyl acetate, 85% hexane) to give the 91 bromide form of a white solid (31 mg, 76% for 2 steps).
Gross formula: CZSH3s4BrNSi.
H NMR (300 MHz, CDC13), c5 (ppm): 7.08 (1H, d, J = 8.4 Hz,> ~, 6.62 (1H, dd, J = 8.4 Hz and J = 2.6 Hz, Pte, 6.58 (1H, d, J = 2.6 Hz, Ph), 2.92-2.86 (2H, m, PhCH CHZ), 2.78-2.73 (1H, m, PhCH ~, 2.68-2.25 (7H, m), 2.11 (1H, td, J = l I.4 Hz and J = 1.9 Hz), 1.73-1.47 (4H, m), 1.60 (3H, s, CH), 0.98 (9H, s, SiC (CH) 3), 0.19 (6H, s, Si (CH) 2).
'3C NMR (75 MHz, CDC13), 8 (ppm): 153.83, 137.61, 131.27, 126.21, 121.96, 119.98, 117.60, 91.38, SI .16, 47.19, 40.46, 40.01, 37.96, 36.00, 30.32, 27.39, 26.84, 26.73, 25.72, 25.67, 22.66, -4.41.
IR (CHCI3), v (cm '): 3052, 2934, 2861, 2238, 1607, 1497, 1468, 1263, 965, 845.
5M (m / e): 473-475 M +, 393 (M-HBr) +, 336 (M-CSH9Br) +.
Alcohol 103 Me ~ H MQ UH Me OMOM Me OMOM
HFI hl FIHFI ti Fv Me0 O 0 HO '~

Ä a solution of tetracycle 87 (28S mg, 0.94 mmol) in tetrahydrofuran (3 mL) to -78 ° C was added 95% ethanol (200 NL) and gaseous ammonia has been condensed (10 mL). Metallic lithium was added per serving until the color blue persists and the resulting mixture was stirred at -78 ° C for 2 h. Of ammonium chloride solid was added quietly until the color blue disappeared and the ammonia has been evaporated. Water was added and the aqueous phase was extracted with ether. The the combined organic phases were dried with magnesium sulfate, filtered and evaporated to give the unstable crude methyl enol ether as an oil colorless (274 mg).
Gross formula: Ci9H2 ~ 02F.
1 H NMR (3 00 MHz, C6D6), b (ppm): 4.66 (1 H, t, J = 2. S Hz, CHO, H ~, 4.22 (1 H, t, J = 8 .2 Hz, CHOH), 3.43 (3H, s, CH = CHOCH), 2.97-2.59 (4H, m, RCHCH CR'R "and R (Me0) CCH CR'R "), 2.26-1.53 (12H, m), 1.24-0.90 (3H, m), 1.20 (3H, d, J = 1.1 Hz, CH).
Ä a solution of the crude methyl enol ether (274 mg, 0.94 mmol) in the tetrahydrofuran (S mL) was added SO% acetic acid in water (S mL) and the mixture has been stirred at room temperature overnight. An aqueous solution of baking soda saturated potassium was added to neutralize the acid solution. The sentence aqueous was extracted with ethyl acetate and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. Ketone ~ iy ~ -unsaturated 102a been obtained under as a white solid (250 mg).
Raw formula: C18H2502F.
1 H NMR (300 MHz, C6D6), 8 (ppm): 4.23 (1 H, td, J = 8.8 Hz and J = 1.4, CHOH), 2.61-1.41 (17H, m), 1.20 (3H, d, J = 0.8 Hz, CH), 1.10-0.83 (4H, rn).
Ä a solution of the crude ketone, tiy-unsaturated 102 (19S mg, 0.68 mmol) in the dichloromethane (10 mL) was added düsopropylethylamine (2.4 mL, 13.6 mmol) and the methoxymethyl ether chloride (775 ~ .L, 10.2 mmol). The resulting mixture has been agitated at the room temperature for 3 hours and an aqueous solution of bicarbonate potassium saturated has been added. The aqueous phase was extracted with dichloromethane and the phases combined organic were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography quickly (2%
triethylamine, 50%
ethyl acetate, 50% hexane) to give compound 93 as an oil colorless very sensitive (155 mg).
Raw formula: CZpH2gO3F.
1 H NMR (300 MHz, CDCI3), 8 (ppm): 4.67 (1H, d, J = 6.6 Hz, ROCHHOCH3), 4.61 (1H, d, 3 = 6.6 Hz, ROCHHOCH3), 4.30 {1H, t, J = 7.2 Hz, RR'CHOMOM), 3.30 (3H, s, ROCH20CH), 2.62-0.80 (20H, m), 1.31 (3H, d, J = 1.1 Hz, CH).
Ä a solution of compound 93 (190 mg, 0.58 mmol) in tetrahydrofuxane to 0 ° C has been added a 1 M aluminum tri (t-butoxide) hydride solution to the tetrahydrofuran (2.9 mL, 2.90 mmol). The mixture was stirred at 0 ° C for 1 h and the sodium sulfate decahydrate was added and the mixture was stirred for 1 h. The precipitate has been filtered and the the filtrate was concentrated under reduced pressure. The residue obtained has been purified through flash chromatography (60% ethyl acetate, 40% hexane) to give alcohol 103 in one inseparable 6: 1 mixture as a colorless oil (173 mg, 63% for 4 steps).
Gross formula: C2oH3 ~ 03F.
1 H NMR (300 MHz, CDC13), 8 (ppm): 4.66 (1 H, d, J = 6.6 Hz, ROCHHOCH3), 4.62 (1H, d, J = 6.6 Hz, ROCHHOCH3), 4.06 (1H, t, J = 7.9 Hz, RR'CHOMOM), 3.87-3.77 (1H, m, RR'CHOH), 3.36 (3H, s, ROCH20CH), 2.26-1.36 (19H, m), 1.34-1.09 (2H, m), 1.06 (3H, d, J = I.OHz, CH).
13C NMR (75 MHz, CDCl3), S (ppm): 129.08, 126.55, 108.58, 106.21, 96.43, 85.69, 67.69, 65.57, 55.24, 47.60, 47.36, 42.40, 42.30, 42.11, 39.90, 38.86, 32.23, 30.64, 29.59, 29.51, 27.68, 27.33, 26.73, 26.09, 24.26, 21.44, 16.75, 16.67 (8 more peaks).

IR (CHCI3), v (cm 1): 3598, 3426, 3052, 2942, 2890, 2836, 1460, 1371, 1269, 1149, 1109, 1046, 954.
MS (m / e): 338 M '+.
SMHIt: calculated for C2pH31 ~ 3F (~: 338.2257, found: 338.2250 ~ 0.0010.
Alcohol 106 and diene 105 Me 9MOM M8 4MOM Me pMOM Me OMOM
--r _ t 'HF ~ ~ HFIHF \' hl F
HO ~~~~ Ph02C HO
1 ~ 104 106 105 Ä a solution of alcohol 103 (6: 1 mixture (a:,! ~) (173 mg, 0.51 mmol) in Ie tetrahydrofuran (10 mL) at 0 ° C were added triphenylphosphine (268 mg, 1.02 mmol), benzoic acid (125 mg, 1.02 mmol) and diethyl azodicarboxylate (161 pL, 1.02 mmol). The resulting mixture was stirred at 0 ° C for 2 h and a aqueous solution of saturated potassium bicarbonate was added. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried with sulfate magnesium, filtered and evaporated. The residue obtained was purified by flash chromatography (10% ethyl acetate, 90% hexane) to give ester 104 (6: 1 mixture (~ 3:
a)) and diene 105 as a mixture which will be separated in the next step.
Ä a solution of the mixture of ester 104 (mixture 6: 1 (~: a)) and diene 105 in the tetrahydrofuran (10 mL) at 0 ° C was added aluminum hydride and lithium (97 mg, 2.55 mmol) and the mixture was stirred at 0 ° C for 45 min. Sulphate sodium decahydrate has been added and the mixture was stirred for 1 h. The precipitate was filtered and the the filtrate was evaporated.
The residue obtained was purified by flash chromatography (50% acetate ethyl, 50% hexane) to give alcohol 106 (6: 1 mixture (, 13: a)) as a white solid (87 mg, 50%
for the 2 stages).

Alcohol 106 Raw formula: CzoH3iOsF ~
1 H NMR (300 MHz, CDC13), 8 (ppm): 4.67 (1 H, d, J = 6.6 Hz, ROCHHOCH3), 4.62 (1H, d, J = 6.6 Hz, ROCHHOCH3), 4.13-4.04 (2H, m, RR'CHOMOM and RR'CHOH), 3.37 (3H, s, ROCH20CH), 2.32-1.59 (18H, m), 1.51-1.27 (3H, m), 1.07 (3H, d, J = 1.0 Hz, CH
).
13C NMR (75 MHz, CDCl3), b (ppm): Decomposes in solution.
MS (m / e): 338 M +, 320 (M-H20) +.
SMHR: calculated for C2oH3, O3F (M ~: 338.2257, found: 338.2264 t 0.0010.
Diene 105 Gross formula: C2pH29 ~ 2F ~
1H NMR (300 MHz, CDC13), b (ppm): 5.80-5.70 (2H, m, CHzCH = Cl- ~ I 4.66 (1H, d, J
= 6.6 Hz, ROCHHOCH3), 4.62 (1H, d, J = 6.6 Hz, ROCHHOCH3), 4.07 (1H, t, J = 7.5 Hz, RR'CHOMOM), 3.37 (3H, s, ROCHZOCH), 2.69-1.57 (16H, m), 1.37-1.12 (2H, m), 1.07 (3H, d, J = 1.0 Hz, CH).
MS (m / e): 320 M +.
SMHR: calculated for CZOH29O2F (M ~: 320.2151, found: 320.2147 t 0.0010.
Ketone a, ~ -unsaturated 110 Me ~ H Mg OH
H
hi FHF
me0 to a solution of tetracycle 87 (227 mg, 0.75 mmol) in tetrahydrofuran ( 10 mL) â -78 ° C was added 95% ethanol (I mL) and gaseous ammonia to been condensed (20 mL). Metallic lithium was added per serving until the color blue persists and the resulting mixture was stirred at -78 ° C for 2 h. Of ammonium chloride solid was added quietly until the color blue disappeared and the ammonia has been evaporated. Water was added and the aqueous phase was extracted with ether. The the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was dissolved in tetrahydrofuran (10 mL) and a solution 1N aqueous hydrochloric acid (1 mL) was added. The mixture was stirred to the room temperature overnight and an aqueous solution of bicarbonate of saturated sodium has been added. The aqueous phase was extracted with ether and the phases organic together were dried with magnesium sulfate, filtered and evaporated. The residue got was purified by flash chromatography (60% ethyl acetate, 40% hexane) to give the ketone a, /. ~ unsaturated 110 in the form of a colorless oil (168 mg, 77%).
Gross formula: ClgH2 $ 02F.
ItMN 1H (300 MHz, CDCI3), 8 (ppm): 5.81 (1H, s, CH = CRR '), 4.18 (1H, t, J =
7.6 Hz, RR'CHOH), 2.52-1.46 (17H, m), 1.26-1.10 (3H, m), 1.05 (3H, s, CH).
13C NMR (75 MHz, CDCI3), 8 (ppm): 199.85, 165.58, 124.76, 108.73, 106.36, 80.00, 47.23, 47.00, 45.90, 45.78, 43.74, 43.45, 42.73, 36.36, 35.04, 27.91, 27.84, 27.76, 27.57, 26.34, 25.26, 25.02, 16.01, 15.93 (6 more peaks).
IR (CHCl3), v (crri l): 3611, 3432, 3016, 2949, 2874, 1662, 1619, 1454, 1364, 1220, 1067, 1007, 952.
MS (m / e): 292 M +.
SMHR: calculated for C1gH25D2F (~: 292.1838, txouvée: 292.1832 ~ 0.0008.
Ketones 111 and 112 Me ~ H Me ~ H Me OH
-. H + H
HFHFHF
p ~ p ~
H

Ä a solution of the ketone c ~~. Unsaturated 1I0 (I26 mg, 0.43 mmol) in ethyl acetate (10 mL) was added 5% palladium (0) on activated charcoal (91 mg, 0.043 mmol) and of the hydrogen was bubbled for 10 min. The mixture was stirred under an atmosphere hydrogen for 1 h then filtered on silica. The solvent has evaporated and the residue obtained was purified by flash chromatography (50% ethyl acetate, 50% hexane) to give the ketone 111 with the AB cis cycle junction in the form of a colorless oil (76 mg, 60%) and 27% ketone 112 trans.
Ketone I11 (cis) Crude formula: ClgH2 ~ O2F.
1 H NMR (300 MHz, CDCI3), 8 (ppm): 4.22 (IH, t, J = 7.8 Hz, RR'CHOH), 2.52 (1H, t, J =
13.9 Hz, RC (O) CH2CH>, 2.31-1.44 (19H, m), 1.39-1.10 (3H, m), 1.07 (3H, s, CH
).
13C NMR (75 MHz, CDCI3), 8 (ppm): 212.42, 108.91, 106.54, 80.48, 80.33, 44.79, 44.51, 42.75, 40.3I, 37.70, 36.43, 35.06, 34.95, 30.10, 28.25, 28.00, 27.89, 27.80, 27.30, 24.42, I9.32, 16.12, 16.04 (S more peaks).
IR (CHC13), v (crri '): 3444, 3019, 2932, 2882, 1706, 1466, 1216, 1077, 958.
MS (m / e): 294 M +.
SMHR: calculated for C18H2 ~ 02F (M ~: 294.1995, found: 294.1989 ~ 0.0009.
Ketone 112 (notches) Raw formula: CISH2 ~ O2F.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 4.23 (IH, t, J = 8.2 Hz, RR'CHOH), 2.44-0.89 (11 p.m., m), 1.07 (3H, d, J = 0.8 Hz, CH).
Diketone 113 Me ~ H Me O
H ~ H
HFHF
OO

Ä a solution of alcohol 111 (76 mg, 0.26 mmol) in dichloromethane (5 mL) have been added 4 ~ molecular sieve (130 mg), 4-methylmorpholine N oxide (61 mg, 0.52 mmol) and tetrapropylammonium perruthenate (9 mg, 0.026 mmol). The mixture resultant was stirred at room temperature for 1 h and then filtered through silica. The filtrate was evaporated to give dicetone 113 in the form of a colorless oil (76 mg, 99%).
Raw formula: C1gH25O2F.
1H NMR (300 MHz, CDCI3), 8 (ppm): 2.59-2.37 (2H, m), 2.32-2.08 (6H, m), 1.88-1.05 (14H, m), 1.12 (3H, d, J = 1.5 Hz, CH).
13C NMR (75 MHz, CDCl3), S (ppm): 219.46, 211.74, 106.80, 104.42, 44.52, 44.24, 42.63, 40.28, 37.45, 36.32, 35.06, 34.94, 33.07, 31.91, 31.83, 30.11, 27.20, 26.09, 25.75, 24.22, 19.10, 12.62, 12.52 (5 more peaks).
IR (CHCI3), v (cm ''): 3020, 2930, 2879, 1740, 1708, 1461, 1215, 929, 746.
MS (m / e): 292 M +.
SMHR: calculated for C1gH25O2F (M +): 292.1838, found: 292.1844 ~ 0.0008.
Alcohol 114 Me O Me O
HH
-.
Fi FHF
p HO
113,114 Ä a solution of diketone 113 (71 mg, 0.24 mmol) in tetrahydrofuran (10 mL) at -78 ° C was added a solution of L-selectride 1 M in the tetrahydrofuran (360 pL, 0.36 mmol). The mixture was stirred at -78 ° C for 1.5 h and a solution aqueous hydroxide 2 M sodium and 30% hydrogen peroxide were added. The aqueous phase has been extracted with ethyl acetate and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained has been purified through flash chromatography (70% ethyl acetate, 30% hexane) to give alcohol 114 sous as a white solid (72 mg, 100%).
Gross formula: C18H2 ~ 02F.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 4.14 (1 H, t, J = 2.7 Hz, 1tR 'CHOH), 2.51-2.40 (2H, m), 2.29-2.05 (3H, m), 1.86-1.I5 (18H, m), 1.08 (3H, d, J = 1.3 Hz, CH).
3 C NMR (75 MHz, CDCl3), 8 (ppm): 220.09, 107.25, 104.88, 66.75, 44.74, 44.47, 41.27, 34.51, 34.38, 33.18, 33.13, 32.13, 32.05, 30.81, 29.26, 26.95, 26.07, 25.74, 24.30, 20.97, 19.60, 12.62, 12.52 (5 more peaks).
IR (CHCl3), v (cm ''): 3611, 3451, 2933, 2878, 1739, 1447, 1266, 1061, 1001, 922.
MS (m / e): 294 M +.
SMHR: calculated for C18H2 ~ 02F (M '' ~: 294.1995, found: 294.2000 t 0.0009.
Silylate 116 Me O Me O
HH
1 ~ ' HFHF
HO TBDMSO
114,116 Ä a solution of alcohol 114 (36 mg, 0.12 mmol) in dichloromethane (3 mL) have been added triethylamine (84 pL, 0.60 mmol) and trifluorometltanesulfonate from t-butyldimethylsilane (4I ~ L, 0.18 mmol). The mixture obtained was stirred ~ the temperature room for 1 h and a saturated aqueous solution of ammonium chloride has been added.
The aqueous phase was extracted with dichloromethane and the phases organic together have been dried with magnesium sulfate, filtered and evaporated. The residue got was dissolved in tetrahydrofuran (2 mL) and a solution of fluoride tetrapropylammonium 1 M in tetrahydrofuran (60 ~ L, 0.060 mmol) was added. The mixture has been agitated at Ia room temperature for 15 min and an aqueous chloride solution ammonium saturated has been added. The aqueous phase was extracted with acetate ethyl and phases combined organic were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (10% ethyl acetate, 90% hexane) to give silylated ether 116 as a colorless oil (46 mg, 92%).
Gross formula: Cz ~ 102Fsi.
1 H NMR (300 MHz, CDCl3), 8 (ppm): 4.06 (1 H, t, J = 2.6 Hz, R.R'CHC? TBDMS), 2.47-2.34 (2H, m), 2.29-2.05 (3H, m), 1.89-1.13 (17H, m), 1.09 (3H, d, J = 1.4 Hz, CH), 0.89 (9H, s, SiC (CH3) 3), 0.02 (6H, s, Si (CH3) 2).
13C NMR (75 MHz, CDCIa), s (ppm): 220.18, 107.37, 105.00, 67.10, 44.83, 44.57, 41.43, 34.70, 34.58, 34.14, 33.15, 32.21, 32.13, 31.10, 29.28, 27.70, 26.07, 25.84, 25.81, 25.73, 24.38, 21.01, 19.73, 18.06, 12.65, 12.55, -4.89 (6 more peaks).
1R (CHCl3), v (cari l): 3020, 2931, 2882, 1739, 1471, 1446, 1252, 1212, 1050, 930.
MS (m / e): 208 M +, 351 (M-C4H9) +.
SMHR: calculated for C2dH4 ~ OzFSi (M '~: 408.2860, found: 408.2863 t 0.0012.
Ester c ~~ -unsaturated 125 Me OTf Me C02Me Me C02Me \ ~ \ + \
Me0 ~ IHF HF
Me0 Me0 ~ 1 a solution of the triflic enol ether 88 (20 mg, 0.046 mmol) in the dimethylformamide (3 mL) were added triethylamine (32 p, L, 0.23 mmol), methanol (233 pL, 4.60 mmol), triphenylphophine (0.7 mg, 0.0028 mmol) and acetate palladium (IT) (0.3 mg, 0.0014 mmol). Carbon dioxide was bubbled to this mixture for 15 min and your solution was heated to 70 ° C under a pressure of 150 psi in a bomb all night. The 18s mixture was treated with an aqueous solution of saturated ammonium chloride and the phase aqueous was extracted with ether. The combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained has been purified through flash chromatography (10% ethyl acetate, 90% hexane) to give the ester has-unsaturated ~
125 as a white solid (13 mg, 81%).
Ester 125 Raw formula: CZiH2sO3F.
1H NMR (300 MHz, CDCl3), â (ppm): 7.24 (1H, d, J = 8.7 Hz, Ph), 6.75-6.70 (2H, m, Ph and CH_ = CRC02Me), 6.65 (1H, d, J = 2.7 Hz, Ph), 3.78 (3H, s, ArOCH), 3.76 (3H, s, C02CH), 2.93-2.88 (2H, m, PhCH CH2), 2.84-2.59 (2H, m), 2.46-2.32 (2H, m), 2.26-2.05 (2H, m), 2.00-1.86 (1H, m), 1.55-1.22 (3H, m), 1.35 (3H, d, J = 3.2 Hz, CH).
13C NMR (75 MHz, CDCI3), 8 (pprn): 164.98, 157.64, 143.41, 137.89, 137.69, 130.70, 127.27, 113.71, 112.01, 108.74, 106.32, 55.21, 51.25, 42.52, 42.22, 40.58, 40.45, 38.20, 38.13, 37.70, 37.34, 30.03, 25.77, 23.07, 14.98, 14.85 (5 more peaks).
1R (CHCl3), v (crri '): 2944, 2565, 1717, 1612, 1501, 1436, 1336, 1243, 1191, 1044, 885.
MS (m / e): 344 M +.
SMHR: calculated for C2lHzsO3F (M ~: 344.1788, found: 344.1779 ~ 0.0010.
Diene 126 (secondary product) Gross formula: CZ ~ H24O3 ~
1 H NMR (300 MHz, CDCl3), 8 (ppm): 7.34 (1H, d, J = 2.3 Hz, CH = CRCOZMe), 7.24 (1H, d, J = 8.8 Hz, Ph), 6.74 (1 H, 4d, J = 8.8 Hz and J = 2.7 Hz, Ph), 6.68 (IH, d, J = 2.7 Hz, Phi, 6.04 (1H, t, J = 1.9 Hz, RR'C = CHCH), 3.79 (3H, s, C02CH3), 3.78 (3H, s, ArOCH
), 2.04-2.96 (2H, m, PhCH CH2), 2.56 (1H, 4t, J = 13.3 Hz and J = 3.1 Hz, PhCH), 2.38-2.18 (4H, m), 1.92-1.28 (1H, m), 1.70-1.56 (1H, m), 1.24 (3H, s, CH), 1.12 (1H, t4, J =
13.3 Hz and J = 3.9 Hz).
MS (m / e): 324 M +.
SMHR: calculated lice CZIHaaOs (l ~: 324.1725, found: 324.1729 ~ O.OOIO.

Ester 127 Me C02Ma My C02Ma HF 'HF
Ma0 ~ ~ Me0 v i25 12T
to a solution of the ester c ~, the unsaturated ~ 125 (15 mg, 0.044 mmol) in absolute ethanol (2 mL) was added 5% palladium (0) on activated charcoal (9 mg, 0.0044 mmol) and hydrogen has been bubbled for 10 min. The mixture was stirred under a hydrogen atmosphere during all overnight then filtered on silica. The solvent was evaporated to give the ester 127 in the form of a colorless oil (15 mg, 100%).
Gross formula: C21H2 ~ O3F.
RMIV 1H (300 MHz, CDCl3), b (ppm): 7.18 (1H, d, J = 8.6 Hz, ~, 6.72 (1H, dd, J = 8.6 Hz and J = 2.8 Hz, Phi, 6.64 (1H, d, J = 2.8 Hz, P ,, 3.78 {3H, s, ArOCH), 3.71 (3H, s, CO ~ CH), 3.05 (1H, t, J = 9.2 Hz, CHC02Me), 2.91-2.86 (2H, m, PhCH CH2), 2.50-2.42 (1H, m, PhCH ~, 2.29-I .78 (7H, m), 1.58-1.30 {4H, m), 1.21 (3H, d, J = 1.1 Hz, CH).
RM1 ~ 1 ~ 3C (75 MHz, CDC13), 8 (ppm): 174.38, 157.62, 137.69, 131.23, 126.80, I
13.69, 111.90, 110.58, 55.20, 53.16, 51.47, 42.10, 41.81, 40.05, 39.92, 31.15, 31.0 ?, 30.06, 29.69, 29.29, 28.96, 26.05, 23.01, 22.22, 17.66, 17.58 (5 more peaks).
IR (CHCl3), v (crri l): 2945, 2862, 1732, 1610, 1502, 1466, 1260, 1199, 1166, 1044, 961.
MS (m / e): 346 M ~.
SMHR: calculated for C21H27O3F (M ~: 346.1944, found: 346.1940 ~ 0.0010.

Alcohols 128a and 128b (proof of structure) Me ÇN M8 ~ OH Me ~~ OH
w +
Fi F
Me0 ~ Me0 Me0 90a 128a 128b Ä a solution of nitrite 90a (4 mg, 0.013 mmol) in toluene (1 mL) at 0 ° C has been added a solution of 1 M düsopropylaluminium hydride in dichloromethane (130 ~, L, 0.13 mmol). The mixture was stirred at 0 ° C for 2 h and an aqueous solution 1 M acid hydrochloric was added. The aqueous phase was extracted with acetate ethyl and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was dissolved in methanol (1 mL) and the borohydride sodium (1 mg, 0.026 mmol) was added. The mixture was stirred ambient temperature for 20 min. A saturated aqueous sodium bicarbonate solution was added and the aqueous phase was extracted with dichloromethane. Organic phases gathered were dried with magnesium sulfate, filtered and concentrated under pressure scaled down. The residue obtained was purified by flash chromatography (30% ethyl acetate, 70%
hexane) for give alkene 128a as a colorless oil (3 mg, 75%) and alkene 128b under form of a colorless oil (1 mg, 25%).
Me ÇOaMe Me ~ OH Me ~ --OH
HF \) H v \ IH
Me0 Me0 Me0 125 128a 128b Ä a solution of ester 125 (3 mg, 0.0087 mmol) in toluene (1 mL) to 0 ° C has been added a solution of hydropropylaluminim IM in dichloromethane (87 ~ L, 0.087 mmol). The mixture was stirred at 0 ° C for 1.5 h and sulfate of sodium decahydrate has been added then the mixture stirred for 1 h. The precipitate was filtered and the solvent evaporated. The residue obtained was purified by flash chromatography (30% ethyl acetate, 70% hexane) to give alkene 128a as a colorless oil (2 mg, 66%) and alcene 128b under form of a colorless oil (I mg, 33%).
Alcene 128a (trisubstituted) Raw formula: CZOH2602.
1 H NMR (300 MHz, CDC13), 8 (ppm): 7.21 (1H, d, J = 8.6 Hz, ~, 6.72 (1H, dd, J
= 8.6 Hz and J = 2.7 Hz, ~, 6.65 (1H, d, J = 2.7 Hz, Pha, 5.21 (1H, d, J = 1.5 Hz, RCH = CR'R "), 3.86-3.75 (1H, m, CHHOH), 3.78 (3H, s, ArOCH), 3.65 (1H, dd, J = 10.2 Hz and J =
7.8 Hz, CHHOH), 2.96-2.90 (2H, m, PhCH CHZ), 2.65-2.56 (1H, m,), 2.36-2.03 (7H, m), 1.75-1.58 (4H, m), 1.17 (3H, d, J = 1.5 Hz, CH).
13C NMR (75 MHz, CDCI3), 8 (ppm): 157.52, 153.29, 137.90, 132.33, 126.55, 116.12, 113.76, 113.72, 111.63, 64.38, 55.20, 51.03, 45.38, 38.38, 34.27, 33.45, 29.97, 27.06, 26.13, 25.63.
IR (CHC13), v (crri l): 3374, 2927, 2860, 1609, 1500, 1465, 1256, 1040.
MS (m / e): 298 M +.
SMHR: calculated for C2pH26O2 (~: 298.1933, found: 298.1928 t 0.0009.
Alcene 128b (tetrasubstituted) Gross formula: CZpH26O2 ~
1H NMR (300 MHz, CDCl3), b (ppm): 7.18 (1H, d, J = 8.6 Hz, ~, 6.72 (1H, dd, J
= 8.6 Hz and J = 2.7 Hz, Pte, 6.65 (1H, d, J = 2.7 Hz, P_ ,, 3.84-3.78 (1H, m, CHHOH), 3.78 (3H, s, ArOCH), 3.63 (1 H, dd, J = 10.3 Hz and J = 8.3 Hz, CHHOH), 2.94-2.89 (2H, m, PhCH CH2), 2.62-1.92 (10H, m,), 1.55-1.35 (3H, m), 1.26 (3H, s, CH).
MS (m / e): 298 M +.
SMHR: calculated for CZOH2sO2 (M ~: 298.1933, found: 298.1928 t 0.0009.

Ketone a, ~ -unsaturated 130 ,OH
Me ~ 2Me Me H
t HFHF
O
me0 127,130 Ä a solution of teiracycle 127 (S mg, 0.014 mmol) in tetrahydrofuran (1 mL) to -78 ° C was added 9S% ethanol (200 ~ L) and gaseous ammonia a been condensed (2 mL). Metallic lithium was added per serving until the color blue persists and the resulting mixture was stirred at -78 ° C for 4 h. Of ammonium chloride solid was added quietly until the color blue disappeared and the ammonia has been evaporated. Water was added and the aqueous phase was extracted with ether. The the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was dissolved in tetrahydrofuran (1 mL) and a solution 1N hydrochloric acid (10 drops) was added. The mixture was agitated at the room temperature overnight and an aqueous bicarbonate solution saturated sodium has been added. The aqueous phase was extracted with ether and the phases organic together were dried with magnesium sulfate, filtered and evaporated. The residue got was purified by flash chromatography (60% ethyl acetate, 40% hexane) to give the ketone a "(3-unsaturated 130 as a colorless oil (3 mg, 75%).
Gross formula: C19H2 ~ O2F.
NMR ~ H (300 MHz, CDC13), & (ppm): S.8S (1H, s, CH-CRR '), 3.77 (1H, dd, J =
10.5 Hz and J = 6.0 Hz, CHHOH), 3.57 (1H, dd, J = 10.5 Hz and J = 8.1 Hz, CHHOH), 2.56-2.21 (6H, m), 2.15-1.71 (7H, m), 1.63-1.01 (8H, m), 1.14 (3H, d, J = 1.S Hz, CH).
RMN'3C (7S MHz, CDCI3), b (ppm): 199.68, I6S.34, 124.85, 110.67, 108.29, 63.69, 50.14, 45.79, 45.66, 43.41, 43.11, 42.78, 36.43, 35.19, 29.58, 29.52, 29.43, 29.25, 26.40, 26.16, 25.29, 23.71, 17.76, 17.69 (S more peaks).

IR (CHC13), v (crri '): 3422, 2940, 2 $ 72, 1664, I4S8, 1364, 1262, 1211, 1035, 883.
MS (m / e): 306 M +.
SMHR: calculated for C ~ 9H2 ~ 02F (M ~: 306.1995, found: 306.1990 ~ 0.0009.
Ketone 131 , OH ~ OH ~ OH
Me. Even H ~ H ~ H
1 '''l''' 1 "
Fi FHFHF
O ~ OO
130 131 prod, secondary Ä a solution of the ketone a ,, li ~ -unsaturated 130 (9 mg, 0.029 mmol) in ethyl acetate (1 mL) was added palladium (0) S% on activated charcoal (6 mg, 0.0029 mmol) and of the hydrogen was bubbled for 10 min. The mixture was stirred under an atmosphere hydrogen for 1 h and then filtered through silica. The solvent was evaporated and the residue obtained was purified by flash chromatography (S0% ethyl acetate, SO% hexane) to give the ketone 127 as a white solid (6 mg, 66%). We also get like product secondary the ketone including the AB trans junction (3 mg, 33%).
Raw formula: C1gH29O2F.
MS (m / e): 308 M +.
SMHR: calculated for C19H29O2F (M ~: 308.21 S 1, found: 308.2143 ~ 0.0009.
Ketone 131 (AB cis junction) 1H NMR (300 MHz, CDC13), 8 (ppm): 3.76 (1H, dd, J = 10.6 Hz and J = S. 9 Hz, CHHOH) 3. S 8 (1 H, dd, J = 10.6 Hz and J = 8.2 Hz, CHHOH), 2. S4 (1 H, t, J = 13.9 Hz, (O) CHHCHRR '), 2.34-1.08 (23H, m), 1.12 (3H, d, J = 1.4 Hz, CH).
1R (CHC13), v (cm-1): 3401, 2930, 2873, 1706, 1458, 13SS, 1271, 1231, 1035, 971.

Secondary product (AB trans junction) 1 H NMR (300 MHz, CDCI3), 8 (ppm): 3.77 (1 H, dd, J = 10.5 Hz and J = 6.0 Hz, CHHOH) 3.56 (1H, dd, J = 10.5 Hz and J = 8.2 Hz, CHHOH), 2.39-1.13 {24H, m), 1.11 (3H, d, J = 1.5 Hz, CH).
Aldehyde 132 OH ~
Me. CHO Me H ~ H
1 "
hi FHF
OO

Ä a solution of alcohol 131 (5 mg, 0.016 mmol) in dichloromethane {1 mL) have been added 4 ~ molecular sieve (8 mg), 4-methylmorpholine N oxide (4 mg, 0.032 mmol) and tetrapropylammonium perruthenate (0.6 mg, 0.0016 mmol). The mixed resulting was stirred at room temperature for 45 min and then filtered through silica. The filtrate has was evaporated to give aldehyde 132 as a colorless oil (4 mg, 80%).
Gross formula: C19H2 ~ 02F.
1RMN 'H (300 MHz, CDCI3), 8 (ppm): 9.84 (1H, d, J = 1.8 Hz, CHO), 3.00 (1H, td, J = 9.0 Hz and J = 1.8 Hz, CHCHO), 2.53 (1H, t, J = 13.8 Hz, (O) CHHCHRR '), 2.27-1.08 (9 p.m., m), 1.33 (3H, d, J = l.4Hz, CH).
Vinyl iodide 137 Me 0 Me ~
HH
Fi F hi F
HO HO
114,137 Ä a solution of ketone 114 (101 mg, 0.34 mmol) in absolute ethanol (7 mL) have been added triethylamine (952 ~ L, 6.80 mmol) and hydrazine hydrate (165 pL, 3.40 mmol). The resulting mixture was stirred at reflux for S days and water was added. The sentence aqueous was extracted with dichloromethane and the organic phases gathered were dried with magnesium sulfate, filtered and evaporated. The residue obtained has been dissolved in tetrahydrofuran (5 mL) and triethylamine (1 mL) was added.
Iodine was added to the mixture until the brown color persists and the solution has been restless for min. Water was added and the aqueous phase was extracted with dichloromethane. The the combined organic phases were washed with a saturated aqueous solution of thiosulfate sodium, dried with magnesium sulfate, filtered and evaporated. The residue got was purified by flash chromatography (40% ethyl acetate, 60% hexane) to give iodide vinyl 137 in the form of a white solid (125 mg, 91%).
Gross formula: CI8H260FI.
1 H NMR (300 MHz, CDCI3), 8 (ppm): 6.06 (1 H, t, J = 2.5 Hz, CH = CIR), 4.14 ( 1 H, t, J = 2.8 Hz, RR'CHOH), 2.62-2.40 (2H, m, CH CH = CIR), 2.19-2.13 (1H, m), 1.86-1.10 (16H, m), 1.08 (3H, d, J = 2.8 Hz, CH), 1.05-0.83 (2H, m).
13C NMR (75 MHz, CDC13), b (ppm): 132.48, 109.80, 105.63, 103.18, 66.85, 45.97, 45.68, 40.96, 40.63, 40.27, 37.36, 37.29, 34.93, 34.80, 33.17, 30.90, 29.28, 27.03, 24.26, 21.13,

20.75, 17.24, 17.11 (6 more peaks).
IR (CHCl3), v (crri l): 3386, 2931, 2873, 1459, 1447, 1380, 1263, 1000.
5M (m / e): 404 M +.
SMHR: calculated for C18H260FI (M ~: 404.1012, found: 404.1008 ~ 0.0012.

Nitrite a, ~ unsaturated 138 Me ~ Me CN
H ~ H
Fi FHF
HO HO

Ä a solution of vinyl iodide 137 (7 mg, 0.017 mmol) in benzene (1 mL) have been added lithium cyanide (3 mg, 0.10 mmol), tetrakis (triphenylphophine) palladium (0) (2 mg, 0.0017 mmol) and crown ether 12-4 (0.3 ~, L, 0.0017 mmol). The resulting mixture has was stirred at reflux for 15 min and water was added. The aqueous phase was extracted with ethyl acetate and the combined organic phases were dried over sulfate magnesium, filtered and evaporated. The residue obtained was purified by flash chromatography (20% ethyl acetate, 80% hexane to 50% ethyl acetate, 50% hexane) for give the nitrite a ,, l3 ~ unsaturated 138 in the form of a dark yellow oil (3 mg) impure.
Gross formula: C1gH26OFN.
1H NMR (300 MHz, CDCl3), 8 (ppm): 6.54 (1H, t, J = 2.7 Hz, CH = C (CN) R), 4.15 (1H, t, J
= 2.5 Hz, RR'CHOH), 2.684-2.55 (2H, m, CH CH = C (CN) R), 2.22-2.17 (2H, m), 2.04-1.96 (1H, m), 1.85-0.82 (16H, m), 1.27 (3H, d, J = 2.5 Hz, CH) Silyl Ether 139 Me ~ Me ~
H ~ H
ti FHF
HO TBDMSO

Ä a solution of alcohol 137 (148 mg, 0.37 mmol) in dichloromethane (10 mL) have been added triethylamine (1.3 mL, 1.85 mmol) and trifluoromethanesulfonate from t-butyldimethylsilane (94 pL, 0.41 mmol). The mixture was stirred ambient temperature for 1 h and a saturated aqueous ammonium chloride solution was added. The sentence aqueous was extracted with dichloromethane and the organic phases gathered were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (10% ethyl acetate, 90% hexane) to give silyl ether 139 as a white solid (190 mg, 100%).
Gross formula: C2aH4oOFISi.
1H NMR (300 MHz, CDC13), 8 (ppm): 6.07 (1H, t, J = 2.4 Hz, CH = CIR), 4.05 (1H, t, J = 2.4 Hz, IRR'CHOTBDMS), 2.62-2.43 (2H, m, CH CH = C1R), 2.22-2.13 (1H, m), 1.89-0.91 (17H, m), 1.08 (3H, d, J = 2.8 Hz, CH), 0.89 (9H, s, SiC (CH) 3), 0.02 (6H, s, Si (CH
) 2).
) tNIN 13C (75 MHz, CDC13), 8 (ppm): 132.48, 109.92, 105.70, l o-3.24, 67.17, 54.23, 53.98, 46.08, 45.80, 41.13, 40.65, 40.29, 37.44, 37.37, 35.11, 34.99, 34.11, 31.20, 29.28, 27.80, 25.83, 25.70, 24.35, 21.17, 20.90, 17.27, 17.14, -4.87 (7 more peaks).
IR (CHC13), v (cm'1): 3019, 2930, 2856, 1509, 1424, 1364, 1217, 1048.
MS (m / e): 461 (M-C4H9) +.
SMHR: calculated for CZOH31OFISi (M-C4H9) +: 461.1173, found: 461.1178 ~
0.0014.
Nitrite c ~~ 13-unsaturated 140 Me t Me CN
H \ H \
-Fi FHF
TBDMSO TBDMSO

Ä a solution of vinyl iodide 139 (47 mg, 0.091 mmol) in benzene (3 mL) have lithium cyanide (18 mg, 0.55 mmol), tetrakis (triphénylphophine) palladium (0) (11 mg, 0.0091 mmol) and crown ether 12-4 (2 pL, 0.0091 mmol). The resulting mixture was heated to reflux for 30 min and water was added. The sentence aqueous was extracted with dichloromethane and the organic phases gathered were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (7% ethyl acetate, 93% hexane) to give the nitrite c ~, Q-unsaturated 140 as a white solid (17 mg, 45%).
Gross formula: C25H4oOFNSi.
1 H NMR (300 MHz, CDCI3), s (ppm): 6.54 (1 H, t, J = 2.6 Hz, CH = C (CN) R), 4.05 (1 Hr, solid, RR'CHOTBDMS), 2.83-2.54 (2H, m, CH CH = C (CN) R), 2.24-2.20 (1H, m), 2.03-1.96 (1H, m), 1.90-0.90 (16H, m), 1.26 (3H, d, J = 2.5 Hz, CH), 0.88 (9H, s, SiC (CH) 3), 0.02 (6H, s, Si (CH) 2).
13C NMR (75 MHz, CDCl3), 8 (pprn): 141.97, 125.37, 115.51, 107.41, 67.06, 52.47, 52.21, 45.28, 45.00, 41.02, 39.55, 39.18, 37.49, 37.41, 34.81, 34.69, 34.07, 31.10, 29.18, 27.76, 25.80, 25.72, 24.33, 21.16, 21.12, 18.06, 14.81, 14.68, -4.89 (7 peaks more).
IR (CHC13), v (crn 1): 3019, 2930, 2546, 2127, 1610, 1504, 1368, 1215, 1168, 1086, 1049.
MS (m / e): 402 (M-CH3) +, 360 (M-C4H9) +.
SMHR: calculated for C21H3 ~ OFNSi (M-C4H9) +: 360.2159, found: 360.2166 ~
0.0014.
Nitrite 141a and 141b Me CN Me, CN
H + H
i 1v HFHF
T TBDMSO TBDMSO
140 141b 141a Ä a solution of nitrite cz ,, l ~ unsaturated 140 (15 mg, 0.036 mmol) in ethyl acetate (2 mL) was added a catalytic amount of palladium (0) 10% on carbon activated and the hydrogen was butted for 15 min. The mixture was stirred ambient temperature for 2 h then filtered on silica. The filtrate was evaporated and the residue obtained has been purified by flash chromatography (10% ethyl acetate, 90% hexane) to give the nitrile, Q 141b as a white solid (8 mg, 53%) and nitrite a 141a as solid white {7 mg, 47%).
Gross formula: CZSH420FNSi.
5M (m / e): 404 (M-CH3) +, 362 (M-C4H9) +.
SMFiR: calculated for C24H3vOFNSi (M-CH3) +: 404.2785, found: 404.2773 t 0.0012.
Nitrite ~ 8141b 1 H NMR (300 MHz, CDCI3), 8 (ppm): 4.05 {1H, t, J = 2.5 Hz, RR'CHOTBDMS), 2.65 (1H, t, J = 6.0 Hz, RR'CHCN), 2.28-1.96 (5H, m), 1.87-1.03 (17H, m), 1.30 (3H, s, CH), 0.88 (9H, s, SiC (CH) 3), 0.02 (6H, s, Si (CH) 2).
ItMN '3C (75 MHz, CDCl3), 8 (ppm): 121.92, 109.58, 107.18, 67.10, 48.46, 48.21, 44.71, 44.44, 41.46, 40.04, 36.65, 36.56, 34.37, 34.25, 34.12, 31.23, 30.85, 30.51, 29.24, 27.69, 25.80, 25.59, 25.46, 21.17, 20.97, 18.06, 16.20, 16.11, -4.88 (7 peaks more).
IR (CHC13), v (crri l): 3019, 2930, 2856, 2240, 1527, 1477, 1426, 121 S, 1048.
Nitrite at 141a 1H NMR (300 MHz, CDCl3), 8 (ppm): 4.05 (1H, t, J = 2.6 Hz, RR'CHOTBDMS), 2.98 (1H, t, J = 9.0 Hz, RR'CHCN), 2.27-0.95 (22H, rn), 1.15 {3H, s, CH), 0.88 (9H, s, SiC (CH) 3), 0.02 (6H, s, Si (CH) Z).
13C NMR (75 MHz, CDC13), 8 (ppm): 120.70, 109.17, 106.76, 67.1, 48:42, 48.18, 44.74, 44.48, 41.38, 39.18, 34.52, 34.40, 34.14, 31.34, 31.27, 31.18, 30.25, 29.92, 29.26, 27.67, 25.81, 24.82, 24.14, 20.96, 20.88, 18.07, 16.66, 16.59, -4.90 (7 more peaks) IR (CHCl3), v (crri l): 2930, 2856, 2363, 1460, 1444, 1252, 1216, 1047.

Alcohols 142a and 142b Me CN Me CN
HH
-HFHF
TBDMSO HO
141 b C-17 = DD 142b C-17 = / 3 141a C-17 = a 142a C-17 = a Ä a solution of silyl ether 141a (7 mg, 0.017 mmol) or silyl ether 141b (7 mg, 0.017 mmol) in methanol (1 mL) was added p-toluenesulfonic acid (Catalytic) and the mixture was stirred at room temperature for 3 h. A solution aqueous of saturated sodium bicarbonate was added and the aqueous phase was extracted with some dichloromethane. The combined organic phases were dried with sulfate of magnesium and the combined organic phases were dried, filtered and evaporated. The residue obtained was purified by flash chromatography (50% ethyl acetate, 50%
hexane) for give alcohol 142a or alcohol 142b as a white solid (5 mg, 100%, each).
Gross formula: C ~ 9H280FN.
MS (m / e): 287 (M-H20) +.
SMHR: calculated for C19H26FN (M-H20) +: 287.2049, found: 287.2045 t 0.0008.
Alcohol 142b (C-17 ~
1 H NMR (300 MHz, CDC13), 8 (ppm): 4.14 (1 H, t, 3 = 2.8 Hz, RR 'CHOH), 2.66 (1 H, t, 3 =
7.7 Hz, 1RR'CHCN), 2.28-1.96 (6H, m), 1.82-1.00 (17H, m), 1.30 (3H, s, CH).
13C NMR (75 MHz, CDC13), 8 (ppm): 121.86, 109.46, 66.79, 48.45, 48.21, 44.62, 44.35, 41.30, 40.02, 36.56, 36.48, 34.19, 34.07, 33.22, 30.94, 30.85, 30.51, 29.69, 29.23, 26.93, 25.51, 25.44, 21.04, 20.94, 16.18, 16.09 (7 more peaks).
IR (CHCl3), v (crri l): 3423, 3019, 2933, 2360, 1523, 1423, 1216, 928.

Alcohol 142a (C-17a) 1H NMR (300 MHz, CDC13), 8 (ppm): 4.15 (1H, t, J ~ 2.8 Hz, RR'CHOH), 2.98 (1H, t, J =
9.5 Hz, RR'CHCN), 2.23-2.16 (2H, m), 2.04-1.06 (21H, rn), 1.15 (3H, s, CH).
'3C NMR (75 MHz, CDCl3), 8 (ppm): 120.68, 109.07, 66.83, 48.46, 48.22, 44.70, 44.43, 41.27, 39.21, 34.38, 34.26, 33.24, 31.33, 31.26, 30.93, 30.29, 29.96, 29.29, 27.00, 24.80, 24.16, 20.97, 20.79, 16.68, 16.61 (6 more peaks).
p-Nitrobenzoate 143 Me CN
H
1v Fi F
HO
142b Ä a solution of alcohol 142b (1 mg, 0.0033 mmol) in dichloromethane (250 pL) have added pyridine (250 pL), p-nitrobenzoyl chloride (6 mg, 0.033 mmol) and 4-dimethylaminopyridine (catalytic). The resulting mixture was stirred at temperature room for 3 h and a 1N aqueous hydrochloric acid solution was added. The aqueous phase was extracted with dichloromethane and organic phases gathered were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (30% ethyl acetate, 70% hexane) to give the p-nitrobenzoate 143 as a white solid (1 mg).
Gross formula: C2 (H31 ~ 4 ~ 2 ~
Mp: 210-211 ° C.
1H NMR (300 MHz, CDC13), s (ppm): 8.30 (2H, d, 3 = 9.0 Hz, Ph), 8.21 (2H, d, J = 9.0 Hz, Ph), 5.40 (1H, t, J = 2.4 Hz, RR'CHOC (O) Ar), 2.68 (1H, t, J = 6.6 Hz, RR'CHCN), 2.31-1.07 (22H, m), 1.32 (3H, s, CH).

IR (CHC13), v (cm 1): 2934, 2856, 2244, 1719, 1280, 1118.
MS (m / e): 472 (MNHø) +.
SMHR: calculated for C26H35C4FN3 (~ a) +: 472.261 l, found: 472.2623 t 0.0014.
X-ray: diffusion of pentane in ethyl acetate.
Ester a ,, Q-unsaturated 152a C02Me Me, CN Me, CHO Me HHH
-.- ,. -HF hl F Ii F
HO HO HO
142a 153a 152a Ä a solution of nitrite 142a (4 mg, 0.013 rnmol) in toluene (1 mL) at -60 ° C has been added a 1.5 M düsobutylaluminium hydride solution in toluene (26 pL, 0.039 mmol) and the mixture was stirred at -60 ° C for 1 h. Sulfate sodium decahydrate has been added and the mixture was stirred at room temperature for 1 h. The precipitate has been filtered on a thin layer of silica and the filtrate was evaporated to give lice crude aldehyde 153a (4 mg).
Ä a suspension of 60% sodium hydride in oil (5 mg, 0.13 mmol) at the temperature ambient in tetrahydrofuran (1mL) was added phosphonoacetate trimethyl (21 p, L, 0.13 rnmol). The mixture was stirred at room temperature for 15 min and the crude aldehyde 153a (4 mg, 0.013 mmol) was added by cannula. The resulting mixture has was stirred at room temperature 30 min and an aqueous chloride solution ammonium saturated has been added. The aqueous phase was extracted with dichloromethane and the phases combined organic were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (40% ethyl acetate, 60% hexane) to give the l3-unsaturated ester 152a as a colorless oil (3 mg, 60% for 2 steps).

Analogues of Digitales Steroids as Therapeutic Agents Abstract: A series of novel digitales steroid analogues, including derivatives containing a 14 ~ i-fluoro substitute, have been synthesized and shown to display binding affinity for the ouabain receptor as well as activity in the inhibition of Na +, K + -ATPase.
Thesis compounds have implications in therapy for a number of medical indications, death notably congestive heart failure, hypertension and cancer. In addition, these molecules can function as natriuretic / diuretic agents and neuromodulators. Multiple methods for the synthesis of these compounds are also presented.
Note on Nomenclature The standard numbering protocol for the positions in the steroid nucleus es as illustrated below. Each of the four individual rings also es denoted by a capital letter starting with A, shown in italics. Stereochemistry es defined as alpha (a), meaning behind the plane of the steroid nucleus (approximated by the plane of the paper), or beta (~ i), meaning above the steroid plane. a-Stereochemistry es depicted by dashed fines as shown at position 3, while (3-stereochemistry es depicted by a solid wedged fine as at position 17.
Ring junctions are referred to as ces, indicating botte substitents attached to the ring junction atours are on the saure side of the ring plane as between the A and 8 rings or trans, indicating substituents at the ring junction atours are on opposite sides of the plane as between the B and C rings. The a- and ji-designations for stereochemistry are applied to the ring junction atours as well.

C ~ 16 \\ '\''' ,, '9 a.m.
14 '~
w H 15 NEOKIMIA, Inc. Page 1 of 31 Digitales Steroid Analogues Patent Applicatiozz January 2003 COMPOUNDS OF THE INVENTION
RSR8 i '' R9 R1o ~ RSR ~ ~ 'R9 RIo , ~~~ m ~ Ril ", ~~ nnRl Rs i ~~, Rs y, Ra H Ra H
H ~ XV ~ ~ H
RiO ~ __ ~ / ~~. / RiO ~ _ \ / ~.
Bz R3 Rz R3 Class 1: 1413-Halogenated Steroids (I) Class 2: 14,15-Q Epoxy Steroids (II) RS _ _ ~ R9 __ ,. "__ mo ,, ~~ u11Ri1 R5. R9 Rs y, Rs i ~~, ,, ~ mt1 R ~ i Ra H ~% Ra H ~ i R'Y ~ ~ 7'R7 '!

t "i Ra R3 R3 Class 3: ~ Ia, is-steroids (III) Class 4: 3-Functionalized Steroids (IV) Wherein for the compounds of formula (I), (II), (III) or (IV):
X represents Fl, Cl, Br, I
m = 0-6, n = 1-6, p = 0-3, each uniquely selected wheréver employed Rl represents H, Me, Cz-Cs alkyl, (CHZ) nPh, (CH2) nPh (substituted), C (= O) Ra, monosaccharyl, disaccharyl, trisaccharyl, amino acyl, dipeptidyl, tripeptidyl, where the Cz-Cs alkyl es unsubstituted or substituted by NR ~ or NHC (= NRd) NRbR ~, where Ra, Rb, R ~ and Rd have the meaning defined below Ra represents H, Me, C2-Cs alkyl, (CHz) nPh, (CHZ) nPh (substituted), where the CZ-Cs alkyl es unsubstituted or substituted by NR ~ or NHC (-NRd) NR ~ and where Rb and R ~ have the meanings defined below Rb, I ~ which may be the sound or different, represent H, C1-Cs alkyl, (CHZ) ~ Ph (CH2) "Ph (substituted), or Rb, and R ~ may form, taken together with the nitrogen atour, a feue-or. six-membered monoheterocyclic ring optionally containing one or more further heteroatoms selected from oxygen and nitrogen Rd represents H, C1-Cs alkyl, (CHz) "Ph, (CH2)" Ph (substituted) Rz represents H, Me, ORe, where Ré has the sound meaning as previously defined for R ~
NEOKIMIA, Inc. Page 2 of 31 Digital Steroid Analogues Patent Application .Janz ~ ary 2003 R3 represents H, Me, ORd, OC (= O) Rd, where Ra has the previously defined meaning R4 represents H, Me, 0-C (= O) Rd, CHO, (CH2) m ORa, (CH2) nNR ~, where Rb, R ~, Rd have the previously defined meanings R5, R6, R ~, R8 which may be the sound or different, represent H, ORd, O-C (= O) Ra, where Rd has the previously defined meaning R9 represents H or Me Rlo represents H, (CH = CRf) p- (CH2) m (CHRf) nA, {CH = CRf) p (CHR12) - (CHR ~) nA, (CHRf) m- {CH = CRf) pB, plus the structures shown below (the arrow indicates the site of the single bond to the steroid nucleus) OOOO
OO
O ~ O ~ OO
Ri Ri Ri R; '~' ~ ~ ""' R ~ R ~ R ~ R ~

NOT
where A represents NOZ, COZRa, CHO, R12, CRAN-R12; B represents NO2, COZR ~, CHO, CRAN-RI2; Ra, Rb, R ~ have the healthy meanings as previously defined; Rf, R ~, Rk have the sound meaning as previously defined for Rg; and Ri2 has the meaning as defined below R; represents H, ORa, OC (= O) Ra Z represents O, S, NR; 2 where R12 has the meaning as defmed following R12 represents ORg, NRhR ;, or NHC {= Y) NR ~ R ;, where R ;, and R ;, which may be tea sound or different, have the healthy meanings as previously defined for Ra, with tea added option that when Rh es H, then R; also has the meaning NHC (= NH) NH2; rg has the healthy meaning as previously defined for Ra; and Y represenfs O, S, NR, where Ra has the previously defined meaning When the compounds of formulas (I), (II), (III) or (IV) can exhibit tautomerism, the formula es intended to include all tautomers. In addition, the formula es intended to include within its scope both (R) and (S) enantiomers for chiral centers, except for those on the steroid nucleus itself, and (E) and (Z) configurations for double bonds Where possible.
NEOKIMIA, Inc. Page 3 of 31 Digital Steroid Analogcres Patent Applicatio ~ r Jafiua ~ y 2003 Figure la: Prototypical Digital Steroids HO O, "~
Me OH OH OH
a- L-rhamnoside Oubain (la), R = oc-z-rhamnoside Ouabagenin (lb), R = H

O
H O-OH

ø-D-digitoxoside Digitoxin (2a, Rl = ø-n-digitoxoside, RZ = H) Digitoxigenin (2b, Rl = R2 = H) Digoxin (3, Rl = (3-n-digitoxoside, R2 = OH) Neokimia. Inc. Page 4 of 31 Digital SteroidArralogues Patef ~ tApplication danuary 2003 Figure lb. Cardiotonic Cardenolides and Bufadienolides H
Bufalin (4) Strophanthidin (5a, R = CHO) Strophantidol (5b, R = CHZOH) H
Actoaigm (b) uzar ~ genm Vin) NEOKIII ~ .tIA, Inc. Page 5 of 31 Digitales SteroidArtalogares Patent.4pplication January 2003 Figure 2: Endogenous Digitales-like Factors (EDLF) H
H OH
19-Norbufalin (8) Marinobufagenin (9, R = Me) Resibufagenin (10, R = H) H
3Gi-Hydroxy-14a, 20: 21-bufenolide (11) Proscillaridin-like inhibitor {12, R = aL-rhamnoside) NEOKIMIA, Inc. Page 6 of 31 Digital Steroid Analogues Patent Application January 2003 Figure 3. Biochemical rotes of cardiac steroids as exemplified by ouabain.
y. ~ cu ~ ..
... ,; . . - ~ - ~ --- ~~. .
-Ouabain action: binding of the hormone to the Na + iK'-ATf'ase pump can activate varïous path-ways. In the events revealed by Aizman et al. 3, signaling is first mediated by the L-type voltage-depen-dent Ca + 'channel and a store-operated channel (SOC). Following involvement of the inôsitol 1,4,5-trisphosphate receptor (InsP ~ -R), oscillations of Ca ~ "result in translocation of NF-xB into the nu-cleus where it regulates several genes involved in vascular physiology. Ca ++
oscillations can also have a direct effect on the contractility of heart and smooti ~ muscle. Alternatively, inhibition of pump function can resuit in local eievation of Ca ~ "in narrow spaces between the plasma membrane and endoplas-mic / sarcoplasmic reticufum (ER / SR). The Na + / Ca * '- exchanger is aiso t: hought to contribute to local rises in levels of Ca ~ ', trïggering protein kinase C (PKC) and AP-1 ~ - or NF-xB ---controlled Bene expression.
Further pathways arise frorn Na + / K + -ATPase interaction with Src, and phosphorylation of the epiderrnal growth factor receptor (EGF-R). One ieads through phospholipase C: (PLC) to PKC; the other through Ras to the Raf-ERK1 / 2 kinase cascade: Ras signaling can influence NF-rcB
through reactive oxygen species (ROS) in mitochondria, whereas PKC can have a direct effect on Raf.
Solid lines show experi-mentally supported pathways; dotteci fines, unconfirmed pathways.
hrom: Scheiner-Bobis, CJ .; Schuner, ~~ 1. IV'atzew ~ olediciz2e 2001,?, 1288-1289.
: '~~ EOIir ~: ~ flfi, Ine. Page 7 of 31 1) ïgïtaJis ~ Stez-oid Analogzces Pate> .zzt Application .Icnzzraiy 2003 Figure 4a. the referenced Structures of the Invention R ~ R1C
HH
R1 = H (13a), aL-rhamnoside (13b), R1 = H (14a), aL-rhamnoside (14b), Ac (13c), (3-D-digitoxosidc (13d) Ac (14c), (3-D-digitoxoside (14d) 02Mc ROC H RrC
H
R, = H (15a), aL-rhamnoidc (I ~ b), stereochemistr, ~ l7a-Ac {15c), (3-D-digitoxoside (15d) R ~ = H (16a), aL-rhamnoside (IGb), Ac (16c), ~ 3-D-digitoxoside (16d) stereochemistry 17 (3 R1 = H (I7a), aL-rhamnoside (17b), .Ac (17c), (3-D-digitoxoside (17d) ~ '~% OKIMIA, Inc. Page 8 of 3l L) igitalis Steroid Analogttes Patent <4pp7icatic> n Janafan ~ 2003 Figure db. Preferred Structures of the Invention O
R, Ric H Ii Ra = H (18a), a.-L-rhamnoside (1Rh), R; - H (19a), aL- ~ ° Lamnoside (19b) Ac (18c), (iD-digitoxoside (18d) Ac (19c), (3-I ~ -digitoxoside (19d) OzIVIc ROC 11 llrC
H
t2 ~ = H (20a), aL-rhamnoside (20b), stereochemistr ~ l7a_ Ac (20c), ~ 3-n-digitoxoside (20d) R ~ = H (21a), a-1, -rhamnoside (21b), Ac (21c), ~ in-digitoxoside (21d) stereochemistr 17 3, R ~ = H (22a), a-1; rhamnosyl (22b), Ac (22c), ~ in-digitoxoside (22d) R'I: ~ OIc1t1 ~ 11A, Inc. Page 9 of 31 l ~ i, ~ italis Steroicl Ancrlogrres Patent Appliaatïon Jarruary 2003 Figure 4c. Preferred Structures of the Invention O
fI
HR ~~ II
RI = H (23a), aL-rhamnoside (23b), R1 = H {24a), a-t, -rha! Nnoside (2 ~ 'b), Ac (23c), ~ iD-digitoxoside (23d) Ac {24c), (3-D-digitoxoside (24d) O {CHZ) zlVMe2 HO O, ter Me OH OH
a- L-rhamnoside R, n CH3 ~, laugh stereochemistry l7çx_ RI = H (25a), aL-rhamnoside (25b), HO
Ac {25c), ~ 3-D-digitoxoside (25) OH
stereochemistr 17 y Itt = H (2Ga), al, -rhamnoside (2Gb), [~ -D-aigitoxoside Ac (2Gc), (3-D-digitoxoside (26d) r ~ 'lïOlil, -i ~ llA, .htc. Page 10 of 31 l7i, critali.5 ~ Steroid Artctlogues Pcatettt. ~ tpplic ~ ation, 7arrztary 2003 Figure Sa. Cômpetition Curve Obtained with Compound 13a at the Ouabain Receptor IC50 ~ 7.1 E-06 M
nH = 0.9 v ° J 75 G
...
G
'Q 50 ...
U
Q) _g _ ~, _6 _5 _ I, og (1.3a] (1VI ~
NIs t) hI _- '~ II ~, Ir ~ c. Page ll of 31 Di ~ italis Steroirl tlnalogtces Patent Applic; atior? Jtzrtuary 2003 h'i ~ ure Sb. Competition C'.urve C) btained with Compound 13d at the Ouabain Receptor - ".
i1H = 1. ~
Lon b ...
. ~.
U
C ~

z ~> ~ y ~ a1 ~ M) h'L ~ OKIMIA, 1190. Page 12 of 31 hi, ~ italis Ster ~ oid Arralog ~ res Patent flPplicatiorr Janucuy 200 -ô -7 -b - '~ -4 Fi); ure 5c. Competition (. ', Urvc C) btained with Compound llia at thc Ouabain Receptor IC: ~ O = 1.2.E-05 IVI
nH = 1.1 100 -t a ~
b .., b G
...
u ~
...
U
U
~, -7! ~ -5 Lc ~ g [16a] (M) . ~% GOhlIt ~ II ~ I, hzc. Page 13 cf 3l l ~ igitoli.s Steroid Analogues Patent Application Jafzuaty 2003 Figure 6. Effects of Compound 13d on Na +, K. + - A.'fPase Acti ~ jity ICSU = 6. ~ E-U6 M
nH = ~ = I .U

. ,.
U
O
U
4-a l ~ og j 13d] (M) : ~ GOkIILIIA, Inc. l'czge l. ~ Of 3.l , '7igitcrlis _Stef ~ oid Analogzres Patent Applicatïort ~ la ~ tzrary 200.3 -ô -l -b -5 -4 Field of the Invention This invention relates to synthetic steroid derivatives and more particularly to IlOVCI
14 (3-halogenated, 14 ~, 15 ~ 3-epoxy, and nt4'ts-unsaturated analogues of digitalis glycosidcs anc! their use for the treatment and detc; ction of conditions associuted with regulation of the sodium-potassium puznp, in particular for thc trcat.mcnt of con ~; estivc bumps into it and hypertension. This invention also relates to rncthcocis for tlzc preparation of thesc compounds.
Background of the Invention Introduction Heurt diseuse remains, as it has every year rince 191 t ~, thc; larintary cause of dcath in North America. In 2000, heurt diseuse causcd almost 1 in 5 deaths in the Llnited States.
(National Vital Statistics Reports) Further. it is cstirnatcd that alnuost 13,000,000 Americans suffer from this would be your diti.on. Thc cost to treat this teller segment is enormous, estimated to be US ~ 12 ~) 9 billion in 2003. (Centers f “r I) isease Control, USA) The fatnily of eardiotonic stcroids (also termcd cardiac or digitalis glucosides / steroids), originally isolated from plants, has been used as phartnace: utical agents for tmore thon two centuries .'- 'Most notabIy, the cc) mpounds ouabain (la) and digitoxin (2a) have bc; en employed successfully for the treatrnent af chronic congestive heurt failurc, whcrc they incrcase circulation through ïrnproved collision: contraction, ~ trvd ntrial fibrillation, wherc they slonr ventricular rate. In addition to thE aglyconea of these agents, ouabagcnin (lb) and digitoxigenin (2b), other representative mcmbers of this uniduc and Interesting class of natt ~ ral products include clig ~~ xin (3) arz ~ l btnfalin (4) as well as tropltantloi.din (Sa) and strophantidol (5b), which like la and lb are highly oxygenated structures.
[dry Figures 1 a and 1 bj ~ hhese compounds at-e generaliy furthcr categori ~ cd as cardcnolicles, tor th ose eontaining ac ~ -lactone in the 17 ~ -positiotz and bufadienolic.les, containing at 173-y-lactonc.
The prïmary pharrnacological proloerty of thcse molec ~ rlcs is their ability to increase thc force of myocardial contraction ïn a dose dependent fashion, terntcd a positive inotropic effect. This activïty has been dczttonstrated to be the result of ïzthibition caf thc cnzytnc Nu + .I ~ + -activated adenosine triphoslol ~ atase (2da +, h'-ArhPasc, IC3. ~ .1.3, sodium pump). 5'8 Na +, K + -ATPase is an itttegral plasma membrane protcin, primarily expresscd in lneart and kïdney, responsible fôr the maintenance of the electrical gradient in all eukaryotic tells, via control of sodium and potassium concentrations. Thesis ion gradients plat 'a key yole in regulating o5rnotic balance, tell volume and maintaining the resting potential membrane. Nà'-coupied transport of 'nutrients, establishment of the iotùc composition of cerebrospinal fluid and aqueous hutnor. electrical activity of muscles and nerves, and receptor-mecüated cndocytosis arc all processes clepc; ndent on thc activity of this enzyme. Inhibition of this ion pu rnp, whici ~ protnotet ,, the inward transport of Na + ions and the outward transport of I <'ion, indirectly leads your year increase in intracellular calcïum ion concentratio: t through mediating confonnational changes to adjacent channels responsible for Na ~ / C; 1+ '' exchangc. I'he subsequent reduction of then of ('a + 2 via the Na ~ lCâ ~~' exchanger lcads to increased intracc: llular Ca ~~ 'z lcvcls and contractile force atrcmger.
, ~ VEUkIML4, lrrc. , fa ~ c:, Jc 15 of 3l I) i ~; ritalis Steroid flnalogaae.s Patont Application .laf7r ~ a ~ y 2008 More recently, sonie of the cardenolïdcs and bufimücncllïdcs, ineluding ltt, 19-norbufalin (6) and its 'Ihr- (~ ly-Ala tripeptidc deuvative,'"
rnarinobufagenin (7), 3 ~ 3-hydroxy-14x, 20: 21-bufenolidc (9), and proscillardin-likc inhihitor (I0), [sec Figure 2 ~
have been identified as endogenous to tnammals as wcll and have been Eoo; ~ tulated to be involved in the normal regulation of Na ~ and K + vïa this rcceptclr. (i Despite their high potency, the digitalis glycosides have narrow tlicral ~ eutic indices and theïr use can often be accompanied by toxic effècts, sonie of wlticlr can even by lethal.
Atnong the most scrious are abnorrnalities of cat-diac rhvtluu and disturbances of airio-ventricular conduction. Indecd, an NIl1-sponsorcd trïal, the I) igitalis Investigation Croup, completed in 1997 showecl a ncutral cffect on mc '~ rtalïiy, although it dicf rccluce thc rate of hospitalization.l2 ulnotller m; zjor detrimental ef ~ Cect ubservcd with digitalis steroids is arterial hypertensïon. t7ther Gomtnon help effccts incladc grrstrointestinal and neurological disorders, blurrecl vision, mlusca, vornitin g and anorcxia.
In particular, the involvement of thc endogenous digitallis flctors in hypertension is an active area of cutrent research. Intereatin ~; ly, titis cf ~ Coca of car <IIAC
glycosides is not directly related to their Na ', k + -A'I'Pasc ïnhihitoty potecicy.l ~ l ~ or example, in contrant to ouabain, its close analogue, digoxin (3 '), dc) es not inducc hylocrtension, although their inhibitory activity is comparable. Further, 3 and cli ~, itoxin (2a) actually reduce the hypertensive effects of ouabain. Irt order tel illrthel 'invcstigatc this disconnection of enzyme inhïbition with hypertensinogenicity, thc hypertelnsive activities of analogs of ouabain were studied and found te :) be sirnilar, but dcfïnitely highcr, to ihc parent in a rat model. Indeed, the hypertensinogenie eff: ect was inverscty correlated with the potency o1 ' enzyme inhibition.i4 Thesc: includcd lb, the dihydro analogue (I 1), ihe ïso-analog (12) and the lactone ring opened derivative (l ~), which in actuall_v a mixture of the aldehyde and alcohol shown.
I-lypertensino ~ enic Ouabain Derivatives ( 'Ozll OIr OIr RCI ~
Düydrn-o ~ abain (I1), R = ai: rhnmnosidc Iso-ouab :: in (12), R = cz-i;
rhnmnoaiy. Ouabain ring-opcncd dcrivntivc (13), R =; xt: rhnmnosidç Z = CtlO, ~ IIZC) ft However, these analogs were only vcry wcak inhibitors of 'thc sodium pump, two "Rders of magnitude less than ouabain and digoxin. 'I hese data demonstrate that the rhamnose sugar and the unsaturatecl lactose appear crüical to the high inhïbïtory potenc.y of ouabain, but do sot appear rclatcd to the abili? y of ouabain tc ~ inducc hypertc; nsion.
r'VL ~ 'OI ~ I11 ~ 11A, Inc. Page 16 oj'31 l:> igitalis Steroid Ancrlo, ~ ues Patent Applicatiurr .Jcnruary 200.3 Further, this suggests that ouabain has adciïtional mechanisms c> f action independent of the sodium pump.
In support of this, recent studies wïth ouabain suggest tlat thc digitalis glycosides are involved in a mueh more complsx serics of biochemical interactions.'S Bin ding of the honrnone to the Na ~ -K + -ATPase pum, ~ can actually activate scveral pathways.
for example, Aizrnan has detnonstrated that c> uabain aces to indues slow oscillations of intracellular Ca + z concentrations and therc: by activatss the transcription fàctor -NF-t <L3.
Since NF-KB plays a yole as a prïmtrry regulator caf ecll activity, lhc ïnvolvement of thesc molecules a signal transduction a.dds a rccw clïmension t "trcir biological actions. ' Figure 3 depicts and describes the carrent understandin g oi ~ ü ~ c; sc multiple rolcs and highlights areas of carrent research as wcll. 'fhc cliveras voles played by ouabain suggest, therefore, that this class af digïtalis steroids may delight; widcr ranging therapcutic potential than has been explored tc ~ date.
Fuuher, Na +, K + -ATPase has been slu> ir-n to cxist in several isoforms that have implications in separation of the toxïc cffects fi-cnn thc pharrnacologically useful actions for these moIecules. ~ 'l9-zs Differsni: ial T ~ esponsiveness in varions tissues 1o the carditonic steroids has been demonstrated as a -iunstion al fcatur ~ c of ths isofonns of the target enzyme. New derivatives, therefore, ~~~ ould likcwisc assist in bettcr understanding ihe pharmacologrical voles of the varions Na'-K + -ATl'ase isofornrs Therapies utilizing alternative positive inotropis agents, cash as ~ 3-adrenergic agonists and phosphodiesterase inhibitors have unique shortsomings of their own. '~''8 Consequently, there remains an urgent nced for a ~ ew thcrapcutic agents which are more specific and effective than those currently ïn use ïn order io avoid the toxic effests. 1n addition, with ouabain and other fàmil, y mc; mbcrs now i ~ cing provcn to bc endogenous species, methods of reversing the cletrimeni: al ef ~ eeas of thcse agents, whcther administered or endogenous, ers higl ~ ly desirable. New derivativcs of the cardia glycosides would therefors bc extremel ~~ desirable tbat woulcl rnitigate thïs toxicity while rctaining the desired biological effects, lviarticularly with rsspsct to positive inoiropy.
Structural Considerations Cardias steroids differ fi-orrr traditional stcroids, apart lï-om thcir biological activïty profile, in three primary structural elements:
~ the sis ring fusion of rings C ~ rD, rathcr than trans ~ the tertïary hydroxyl and C 14 ~ the (3-configuration of the C17 substïtuerct, which is ash: ally thermodynamically less stable The specificity of the biological action of these matsrïals has been ascribed to the speeific thrse-dimensional arrangement caf the substitucnts based upon the unique structural fcatures of the compounds.z ~ ° 3o Repke and coworkers, through studios on a wide variety of steroidal compounds, concluded i: hat it ia actually only the cyclopentanc-perhydrophenanthrene carbozmusleus that is thc noinimal digitalis pharmacophoric moicty.3 ° '3 ~ The particular topi:> logy provided by the 5 (3.14 (3-androstane-3 (3,14 (3-diol ~ VIJ '<) hltl ~ llA, Iras. Pcrge 17 of 31 Digitcrlis Steroid Analogues atertl Application Jauuary 2003 skeleton is reduired in order to interaci. optimally vvitlt tho recoptor anal must bc considered as the basic backbonc ~ in thc; cicvclopaorcnt of any t ~ ev ~
analoytcs.
This importance is emphasiLCCI by a numbcr ot ~ cailmr observations and studios, In digitoxigenin, the convoi ° sion of ~ the trsual C ',, ~ 1:') cis stnrcoc ° l ~ enristry to trans cornplc; trly eliminates inhibitory activity. This is l7ostulated ta occur rince the structure is now more cxtcnded and the lactone moict: y bel; ins tc »nterfcrc witl ~ an ~ iru ~ acid residues in thc key recognition ~. ~? In co ntrast, rnodifi ~: .uti "n oi ~ thc nil3 rïrrç;
jutiction to crans frorn cîs as ïn uzarigenin {7), results in only o rnim ~ r lo: a of, tctivityl, about 3-ibld. ~~
U5 5705531 described perhyclroindcnc derivatives (; 14-15) desigried as sirnplified analogues of the cardiac steroids, which pr ~ aecve sonrc caf thc rnost distinctive fcaturca of tHC; scaffold, but essentially deletc the ~~~ rtnd 13 ricr ~, s.
s Vit, Rt , .s Mc ' / y R-(~ H 'OI I
(m) (I s) I-towever, these cornpounds managed to attain anly tr ~ cderatc bïn ding affinity with all IC; o> (pM, although sonie did noticeably lc. ~ Wer systolic bloocl pressure (SBP) in a spontaneous hypertensivc rat rnodel. 1n addition, extcrrsivc stuclics have becn conducted on a vide series of hydroindene dcrivatives vvith sirnilar results. ~ 3- ~ r 'rs part of this approach, cycloallçyl or phenyl ring suhstitucnts rvc.rc adcled to thc indcne in an attempt to compensate for the loss of the A and B ritys. 'l'ire deriw, atives with the lughest affinity in this compound class had extended arms vlith basic subslituc: nts ïn the Y and Ri positions in analogy to thc situation observed in the parent digituli: -; skc ~ lcton (vic ~ F ~ imfior). THC
di ~ italis analogues. With the fu (I earhcoro slkeleaon, possessecl SO-100x greater binding aflinity, however.
NI-Iz i N ~~, NOZ
.r 11'1 e "- 1: ~ '1-1 z M c HZN N ~, / ~ ~ I ~ 7N ~~~ .i N ~. NOT
Nli, (16) _N1I, lb'EOKI ~ 4IIA, Irzc. IS of 3I Page Di, t; itali, .s Stey ~ oicl Az ~ ulogzzcs Pater! Ap ~ licatiofr Jcrzzzmry 20113 A striking difference which il; ustrat ~ s again the cri.tical nature of t: he C / D
ring conformation is provided by cc>nsidcr; ntion of thc Scx- ancl 5 ['a-iscmicrs in this scrie; s, cquivalent to the C14-positio ~~ in th ~ J dil; ü ~ i.lis stc: ri'ricis ~. In all cases, the latter were 25-30 tunes more active. Ycrhyciroindcnc d ~: rivatïvcs rccluirc: thc 5-wbstituenl to be in the equatorial position in their lowcst enorf ~ y conformations, whiolr for tlte 5 (3-isomer is analogous to that in the digïtalis teroïcls. In c <7ntrast, the Srr, -from = rivals loresent to non digitalis-like conformation of tlm put, ttivc, C / I-) rings. I? Veo whcn thc bcst basic replace groups from the digitalis scrit: ~ wcrc ïr ~ corlnorat ~: d ïotcs tlte; se hydroïndene derivatives, the activity remained at lcast l tI-I «ld below tint of digitoxigcnin. 'FHIS
strongly suggests that the intcraciion of t.he loasï ~ .: anrinc witlt tloc rcceptor is not strong cnough to overcome thc loss o! ' r ~ flinil.y causf ~ d by tlnc hcrhyclroindc; nc skelcton in comparison with the stcroid nuclc ~ .w. 71 '~ is cr »phasizes its importance and poïnts to the rather strong hydrophobic interacaion cil 'tl ~ c: rel; a tivcly rifTicl and <lefined shape of the 53.14 (3-androstanc tore to thc rcccptor.
With the requirements for the cntirc skc: lc, ton rccof; nizcd, tire next kcy cansidcration is the substituents at the various sites aro vnd tl ~ e ring. l ~ irst, if is clcar that tHC; high number of polar groups arrayed around fhc steroid m.ac.leus as cxlrihited hv la, Sa and Sb is not considers. The other most i ~ otal> le mernbcrs of this fnmily, 21a and 3 are rnuch less Functionalized. For the digitralis steroid:;, small moditicatioets in ean lead to structure dramatic changes in biological activity. .- 'gis an exatuhlc, a atereoisomer of ouabain with a 1 a-hydroxyl (17) dïsplays higher potes cy than 1 a and: ~ as a positive inotropic agent (WC> 97f19099). Further, thi ~ ,, conipouncl han,; in iml ~ rov ~ d tlrc.rapc: ntic ratio with respect to toxicity associated with induccd arrhytatmi; is.
hc o yes W () 97/1 ') 099 (18) Similarly, dihydroouabain (11), has an equivalcnt tlscrapeutic effcct to ouabairt, albeit sot pick as strong. It does, however, possess? X the therapeutic wïndow. (GB
949 407) ~ lost of the research efforts in this arca have becs dïrectcd towards modification of the 1% ~ 3 position. In thc; natural products, tltc unsaturated y- and b-lactose moieties are : ~ '~~ Okl ~~ flA, Irrc. Page 19 qf 3I
C? Igita, lis Sternid Analogues aterrt Application .Icrrzzrary 2003 associated with high receptor binding aflïnity. 11 shpul ~ i be ~ toted that the bufadicnolides, such as aL-rhanmosyl-bufaliu, cau exhibit itihibitory Itot.cuciw an urder of magnitude above that of ottabain, 1.5 ttM versus _ '1 nM9.z ~ tes alrcac.lv allucled ta, s ~ tl: uration of tlte lactose ring in botte la and 3 lcacl ta corttpounds witlt lov-cr ütlibitory activity against Na + -K + -ATPase, but which Itavc; crthcn latvorablc bioploysical propertics, such a5 bcin, g rapidly washed fiOt71 fabrics. Wc) 0 () / ~; 76L4 dcscribes a dihydrc ~ ouabuin-liter factor, also containing a saturated lactose ring, with irnlribitory activity 70 ~: 1c> wcr {I (~ ',, o = 5> 0 nM) than ouabain-leks factor (1C {~ ~ - ECU nM). but 3x higlicr than 'i 1 (1C; o -= 1.7 p M) itself:
More significantly, investigation of the rcpfacen ~ ent of thc lactose with a furan led ta the dïscovery of a clinïcal c.andülate 2 ~.> u tlrc treattncttt of hyloertension, PST 2.238 (l '' - 3-furyl) -5 (i-androstane -_ '3 (I, 14 (i, l7a-trial {l9). ~ l' Phis and other such ferras derivatives are clescribed in 1JS 54321 C9, 5591 734 and 5593'v) ô2. the ST2238 was selectïve for Na +, K ~ -AT'Pase ovcr other rcccptors ancl cnL ~ tnc: ~ involved in blond pressure regulation and hoi-tnonal control. It ïs a ouabain ant ~ t, ~; oarist, ~ with as <> tltcwhut high IC, ;; o of 1.5 LtM.
I-Iowever, and binds sigmificontly fastev tl ~~ lll Oltahilitl ta thc receptor.
Further, this derivative was orally active in rcdttcin ~? ltr blond systoles; ssttrc (S13P) in rat tnodels of hypertension, with an ECS ~ = 4 ltgikg. lm, ~ ortantly, 1 ') also did sot display detrimental cardias effects typically associatcd wiTb tllis fïtrnily. Iutrt (er, ït: es devoid of diuretic activity and its associated ride eftects. 'hhis ccnnl ~ onnd ïs tkte first treatment that acts boot as a ouabain antagonist attd 1o cc ~ urttcroct itoluccd atncl gcnctic hypertension. Aces, and pas become the ftrst syuthctic analog of rod cardias elycosides ta reach humas clinical trials and es currently in I'ltasc If testing 3 '' - 'r1 wT
I
HO HO
you PST2238 (19) (2U) Another heterocyclic moiety that Iras becs utili ~ c; d in the l 7 (3-position with positive e.fféct es a pyridazine (EP 583578) .x 'Substitution ~ .of the hetcrocyclc ior the lactose in digitoxigenin (2b) yielded the analog 2 (1, whiclt Itacl a l. ~ ir ~ ding aftinity close ta that of the parent f ~ ICSt ~ = 0.10 AMI versus 0_0f> 3 pM). In tlùs system, the Corresponding 17 (3- (3-furyl) derivative, in analogy ta P ~ aT 2238, but lacking the Cl? A-hydroxyl, gave an ICSr, of 0.25 ~ tM. It es interesting ta note that a foras was actually used as the synthetic precursor ta the pyridazinc.
; VBC) hZ ~ LII ~ l, Inc. Page 20 Digital Steroid Analogues Patent Alplicatiorr .IanZrary 2003 Apart from the lactonc derivativc s iouncl itt thc n Gtturttl l ~ roducCs and thcSe heterocycles, a number of other groupa loavc bc.:en re-read> rtuct to be, ~ rble to bc substituted into the Cïl7 position whilc tnaintainin g c> r cm, n enhaucirlg tlaa tm.tivity. in particular, fhose moieties that can generate a positïvc charge hase prcwen rnost c.hlëctivc, undouhtedly due to a favorable interaction with a ncf; al ~ vcly charf ~, ecl reaiclue or site of the receptor. A serïes of 1.1S patents (IFS 53? .471r) s444tt55, 5 ~ '.ïh <> ci0, _i5fi76 ~ 14, 518312.7, 5599806, 5705662, 5792759) and relateci scientific peblicat; uns ~ '~ -'t ~ dcs ~: rihc ~ s thc, sc;
cflvôrts. ln particular, US
5444055 detines digitarlis stcrc »d ~, contuioiny, ono or trtore oxitrt ~
naaieties. These were shown to have vert 'strotag al ~ fïraity for tlfc ~ c> ualaain rcc ~; lrtc ~ r, below 100 nM in many casus, and significantly lowcrecl ~ 'sl3l' 1c: tluu in river rai tnodel.
Ijnportantly, this effect was not aecompanied by <rny antiarr ~. ~ thtnïa. '1'lrc mc> st active compounds reported to date, however, contais an amint7allcyloxinw ntoicty in tlrc l'7 ~ 3-I ~ ositiotl (CJS
55831? 7 5599806, 57056fi2). ~~ 'hhe bindiog afienities of the bort moiecules from an extensive series synthesired werc: ahproxirr ~ atcly '? -_' i timcs rnurc potcart Chan digoxin. Further, many of thcse 17 (B-analoyue ~: have becs clcrnonslratcd to laavc improvcd safety profiles ao s ~
over the natural cardiac stcroids.
The other situ in thc digitialia ste: rcuid cors tl ~ at has bcen studïed rxtost extensively 1s the -position. It has been dcmonstratcd that thc 3 (3-hydrc> xyl 1s sot essential for activity.4 ~ ° 53 For example, the 3-dcsoxy dot-ivatsve stikl exhïbïts <tn IC'Sf, of 7 () nM
against the bïnding of [~ H] -ouabain to ils reccL ~ tor. tlowcver, thi s 1s 3-fold when than the 3 ~ 3-hydroxy compound and an order r ~ t ~ magnitude ~~ elca ~, v those coml. ~ otmds wïth a sugar moiety.
Similar to the investïgatiorrs jcrst de ~ scribcci f «r the C1 7 positicm, a number of dcrivatives ivïth a variety of basic tnoictics at the (. ': ~ position have also been synthesized and tested (C155444055, 5478817, 5489587, 5538c? 60, 556'76 ~~ 4, c5384? 50), but with much then bcneficial effects on biological potcncy btvin ~ observed.
Further studios have shotvn that the 3 (B-saccharifie 1s sot an esseritial element for thee cognition. f Iowevcr, glycosidation has becs wcll-ostahlishcd in laroviding benefits in Print <svinb soluloility, tnetai ~ olisn ~, tüstribution ancl other lharmacokinetic properties of tlcesc molecules. ~ 3 Vvittr rcahect to thu saucharidc portion of the molecule, ci.-L-rhatnnosides seems to provide i.he must acctive conrpounda, although (3-1 ~ -glucosides are comparable. lu the cardenolide>, serves cnuch diftèrc: ncc. 1s observed between these and the digitoxosicies, howevcr in 4raalo! _; ucs ~~ 'ithuut a 17 (3-y-iactcute, ttrc order ofpotency ïs nuore pronounced; K
a-t.-rhamno wrinkles> (3-n- ~; lu ~ .osides> digitoxosides a'l -OKIII ~ TLfl, hic. Page 2 I o ~~ 31 I ~ i, ~~ ilctli, r Steroid Analogx ~ f7s l'rrlc> nl. ~ Ipplrcvation Ja7nunry 2003 ! ~
Mc 21a, lt = a-1, -rhamnoside, lC ~ o = 3 nM

21 b, Il ~ (3-1 ~ -glucoside, lCSa = 7 nM
21c, R = naanodigilcuxoside, ICSa = 9 nM
21 cl, R == biscligitoxoside, ICSO = 8 nM
~ 21 c, lt ~ tris <ligitoxc> side, ICim = 8 nM
21 f, 1Z ~ iJi-I, IC'so ~ 24 nM
2I h, R = II, I (. 'Stt ° 70 nM
I ( L.astly, it should be noted that pi ~ tcncy is not aflè.cted hy addition of the second and third digitoxose units. Howcvcr, thc sacch.n-idu cran have an c; ftect on activity as well. for cxamplc, with 1 d (3-hydroxyprogcstcrcrnc, tire (iU-glucosidc was approximately 10 times more potent thon the a ~ lyc: orre. ~ "t I ~ t ~ -thc_i, unlïkc oual> airc, which potassium enhances excretion witlt little ei: fect con scrcüum Icvcls, thc glucosidc exhibited strong natriuresis, with little effect on lrotas:> ium. ~ '7'iùs is consïstent with rc = sults obtaïned for othcr tnembers of the digitaloitl prcgnane family, including L ~ iD (~ 23 (-vide itiyu) Of the other possible sites ou lhe digitalis steroid ccrre, the only other that lias been shown to have any specific effects on bindin ~~ rtctivity and biological potency is the C14 ring junction. rI "hc cffect of thc conlil; uralio; r at thal l ~ osilior ~ is dramatically illustrated by the differcnces in cardiotonic activities coi thc S ~ i, l ~ l (3-pr ~ e ~ gnanes.S ~
the original cornpounds with a trousers C './ l :), junctit> n arc esscntially clcvoid of inotropic action, ~ lycosides of thc ~ (3, t4 (l-hrcgrranes exert p <~ sitive inotropy. In addition to tea configuration, the actual substituewi _rt tire. C ~ 14 position alao makes a difference.
Introduction of ai 4 (i-hyydr-o, ~ .yl t; a lo-ogrsterona has a marked effect tus 141-hydroxyprogesteronc exerts a pe ~~ ïtivc imutrcrlaic effec: l missing from the parent hormone. More importantly, tlrcsc pregnunc di; ri ~ ratives have a scEperior safety profile.
Othcr substitutïons at this position also pan lrc porf ~> rmed with retention of activity. hor example, the replacement oh tire 14 ~ -hydc ~ "xyl with an arnino tnoiety as in 14-amino 3 (3- [(çx-L-rhamtlosyl) oxy] -5 (~; 1. ~~ 3-pregnane (I.; NI? Fs23, 22) resulted in a compound of equivalent potency to ou, tbairr.s '' s '' lndec: d, 22 had lctvorable enough characteristics to be eonsidered for clinicat investié; atic.nrs rv'EC? IütLIIA, Irtc. Page 22 of 31 t ':) igitali's ~ S'ter-oid Analogtre, r Patent Alrhlication Jant ~ nry 2003 ~~ r ' OII
1n contrast, a soies of 14 (3-mctlaoxy cte: ivatives showcd considerably reduced binding atftinity.4 ~ Contrastingly, tltc rtaturul t ~ rc> duels rrtarïnobufàgenïn (6) and resibufagenin (7), which contain a 14.15 (3-cpoxiclc us a cüstinguïsl ~ iry; fcaturc tlrat liras been Dernonstrated to hc att important factor in thcir biol_yical activity, cxhibit higlt aifinity binding.z '~' 6t This st.tggests that thc rcclueed binclin ~; with thC 14j3-meihoxy was likcly not due solely to the loss of the hydrogen bond donatin ~; a: bility o1- the hydroxyl. Instead, the results cletmmstratc tltat thc receptcor has a strict aic; ri ~~ rcdt ~~ irement at that site.
From the reported activitics with C, t4-substituents, it appcars that what is desirable would be an entity that duplicated thc hydrogen honcl acceptor activity of thc hydroxyl, but was small în overall size, hortunately, fluorir; c maichcs that description.
Lndeed, the unique propcrtïes of this rocnrber crf tl ~ c pcric. ~ di ~ .- table makc it extremely effective for ntodttlation of 'bïological activïty. ~' ~ 5 ~ hc i-luo rinc: atour is approximately the healthy site as a hycirogen, but thc dranratic diftèrï: n cc: in clcctrortegativity oftcn modify the clcctronic nature: off thc resulti.n ~; :; tructurc aod hcnrcc nzï>dilïc; s bicological actïvity. in partic_ular in the stcraid fcld, iniz ~ ociuctioz ~ of lluorinc in the; C ', (~ c> r C; 9 positions has been founcl to enhancc physiologie,: al acti ~ ity cony.zarcd wiilt the parent structure.
~~ -I ~ luorinated derivativcs oF stcroids Ira ° ~ c ~ t, cc; nw ~ ..ll-cstatblished to exhibi.t cnhanced <Totivity. For cxatnple, lalotesti ~ a, 9a- (luor <a - 1 1 (i, 17 (3-dihyctroxy-17a-methyl-androst 4-cn-: 3-one is a Notent, androncrgica lly active compound.
tJS 42 ~ 34 ~~ 8 and US 403Ci8 (ï4 dcscribe steroidal compounds possessing a 14a-fluoro replace and relate: d synthe-tic. preacesses. Ctf partïcular interest were a4a-fluorotestosteronc, 14a-fluoretprogestcronc and 14a-fluoroandrost-4-ene-3,17-dione.
'I ~ hese cotnpounds arc of'ïnte; rcst sincc tltcv exhibit thc sain general biaactivity, androgenic, progestational and atnclrogcnic respectivcly, as the parent unfluorinated stcroicl, but at an itnprovcd lc.vel. l ~ urtltc; r, hc> tlt l4cr-fluoroprogesterone and 14a-fluoroandrost-4-ene-3,17-dïonc displ, .iy an enhancccl level of activity on oral administration in comparisou ivith tltt Lorrespondint; l4cx-hydrogen compounds.
'1 hese reaction methodolotries utili-red electrophilïc fluorination terms (trifluormethyl hypofluorite, rnalectilar Iluorine) ihat lead cxclusively to the alpha stcrcoisomer. 1'1te conditions werc establishcd such that free radical fluorination was supprcsscd. T'hc l4jl-ïsonrer was lzreclucied ttu ~ ough the directin ~; effects of the other h7 ~, OklitllA, hw. Page 23 of 31 1> i, ~~ ilalis Steroid Attalo ~ ues l'ctlcnt.flhplicatiotr Jamaty 2003 L, JV I) 6L, 5 (ll) suostituents on the steroïtl rtttcleus which té ~ rce tais reatctiora to hroceed with retention of contïguration. 'Plus ril; orous st ~:, reosp ~: cifïcity uf the; electrophilic fluorination reaction was indepetdently contïrnrc-1 by Rozc; rr an <i k3en-Shushan. ~~ '' ~ '~ In each of the illustrated ~ a. <; es, with ~ lmlcstcrc ~ l, c ~ lmolt ~ stwv-71, an ~ f uh ~:> l ~ rnic acid derivatives, the reaction to install a tluorïnc ai ihc 14-positi <~ t ~ ci ~ d le; ~ cl tzr sïde products. These were sot tea l ~~ s-stcreoïsorrrers, but ratlncr the I'lcc-rc: gioisamers .. 'fhe stereochemistry was unambi ~: uc> usly assigncd ilwough r ~~: -NI \ ~ Il ~ mherc; the ehernical boot shifts and coupling report: s crf tlae y-carbcw atesms v = ar-v si ~ mitïcantl, y dehenclïntl on whether they are asti or left to thc fluorite Gitom. ~ '~ L: rstly, 3-t) -trc.ctyl-14-deoxy-14a-tluorodigitoxigenin has becw preparcd in lorv yiclcl tiwrn ~ loer cor-rcspondirtg 14 (3-lrydroxy derivative by treatment witfz anhyclrcrus lCl ~ and St ~ Iy. ~, o > Iovvever, exploration of thc iccy C '14-position with a fluorine substitute has sot yet been addressed with the samc bcta stcrcochcnrisiry as thc natural product required for optimal hiolot; ical activity. 'l'lre or-ny crm ~ pcn; r ~ cls r-cpor-tod to date containing a 14 (3-fluoro urhstitucnt arc thc thrce prcgnanc- i '1,20-slionc: e: leriv; ctives shows (23 ~. ~ °
"111 c t t) R
23a, R = H, 16a 23b, R = Ac, 16a 23c, R = Ac, 1G ~ 3 v ,,.
Ac () ~~
II
lr ~ this report, the cornpounds wcre c> btaincd a: ~ tlrrec among six components of a mixture rcsutti.ng icom the opcning ofthc 16., 1 'la-cpoxy cler-ivative mith 1-IF. Tea alcohol 23a was ihe only ose of these threc obli: tto hc isoiated alter careful column chromatogra, phy, goal vidais znerelv used as a ~ rynthc; ti ~: intcrmcdiate fcar further transformations and no biololyical activity was invesügatcd r> r repc.rrtecl.
.h'Lt) IiIrLfl_A, Irrc. Page 2. ~ of 31 there ~ Jiralis Slenoid Araalo, ~ rros a ~ cn ~ Application January? 003 I) esc; rlptlon (? F thC Invention ! vIo (cc; ulcs oC thc: invention have: Recta dasigned and constructed ta build upon the c>hsc; rvatiotts outlined in the previous sec; fion c3n tha actïvety profile of the cardiac ~ Lyc.osidcs. '1,1-vc invcntiotz providcs alter-ttative methods ta crcate navel structures your fill t; ap; ~ in l.nowlcc.igc for this thc; rapautically important family oC
cotnpounds and determine if are i "~ hrovud laiological prolïlc v ~" itl- ~ bout potcrzey and safety can be attained from them.
~ 'Ynihësis rl "hc navel stereochemestry attd complex structuras of the digitales steroids, PARTICULARLY
thc ntorc highly fitnctionalü; ed tnanth,:; rs, rnakc thesc compounds challenging and inturcsling synthctic iargcts. Cieneraliy, approaches ta analogues of the digital t; lycosides lzatva hecn lht-ouglt transformation <> 1 ~ other members of thïs class of cotnloetunds.?''"3°''~-'~; Inclccd, al, irl; e lunthcr of derivativas, such as the analogues outlined in lite; previous section, liane heera invcstigated in the search for cardiotonic cligitalis-lilce cotnpouncis witlz motu .favrr ~ ablc toxicity prc> perties.
Surprisingly, despite tlzeir long mcdicinal usal; c, fi = .wc / c rnovr ~ syntbetic efforts have l ~ een directed towards TLTI; is; enteresting structuras irn orclvr ta ttvodulata activity crf the natural materials. AT
~ aric ~ iv of synthetic stratcgics havez. heer ~ aituad a1 the digital skeleton, ~ 4 ~~ 5 however, cleslaitc tlzis, thc hrst total synthcsis of a tTZeanber of this famïly, (-i-1-digitoxigenin, was cznly z-elativcly raccntly achie; vcd. ~ r ' Spccific synthelic clclails and reaction ;; chcntes for representative compounds of the Iorcvrnt invention arc presented in the attachcd l'l ~ .D. thcsis, "Synthesis total of 14 (3-vlttorostéroidcs via lu rc; action of Dit.; Ls-Al <ier transatmulairc "..
which es included as ~ ttn integral part of this application. As o ~. ~ Tlined praviously, the cardiac steroid family es cliffc; rentiatad ft-ont the mora traditional s ~ i; roi <l lto ~ -tnones by ihc ces ~ junction between it ~ e C 'amd I) rings; a fcaturc tlzat also provid ~ s otu ;; of the greatest synthetic challenges.
'fhc i irst ahproaeh ta the conslructeott of thc. nonce utoleculcs of the invention connect on thc Ioowarful transannular Dials-Aldar (I "AUA) reactüm discovered and developed within the laboratorics uf on.e of thc irtvc; ntors.r 'l ~ or comloc> unds of f ~ lass l, application of the '1 ~ AI> ~ 1 3br t.hc first lime ta a tluoroaikenc. ha ~. pertnitted absolute control of the s; torcochezzzistry at tlte kcy GiIO ring jonction ta provide only tlzc desired 14 ~ fluoro substitute your proparlv tttïnlic tha confi (~ uration of tha digitales steroid Family. Year alternativa meihodology es also provided tlat pal-mits the introduction of the 14 (i-fluorite t'r <> m rod alrcady assernbled steroid skelctosz.
In this strategy, thc 143-hyciroxyl of a protccted digitoxigenin derivative es treated with dicth5-laminosulfur trilluoridc (DAS'h). In catttrast ta thc conditions employcd by Haas anci Kortttlann ~ 9 in which only a lcwv yïeld of the 14a-fluoro compound was Obtained afonl; with considerablc antounts of sicle products, this approach from essentially the saznc prc.cursor suprisingly provided thc ~ ld / ~ -fluori> analogue exclusively.
similar mcthoc.is ara re.hortcd fc> r the conversion of thc 14 (3-hydroxyl enta the other halogenated clcrivativcs of this C'lass.
1n additïon ta their direct aphliaation as tharttpetttic agents, molecules of Class 1 can be uscful as syzuthetic intermcdiates towards thc construction of other new steroid Nf ~; OR7u11A, Irtc. Page 2_5 of31 l ~ i ~, rilali, s Stenoid ~ lnctlo ~ lrtc ~ s l'cttortt f9plolrcoztiorr Jcmttcny 2003 dcrivativcs. ~ sidc rcactïon obscrved iv their preparatïon can lead to the Corresponding climinulicm products of Clans 3. 'I'hat sarruc transformation can be purposely executed to >> ic ~ cl thcsc unsaturutccl derivativus. 'l'te rcsulting alkenes can then in turrz be used fox corvversion to cc> nycout ~ d :; of 'C'lass 2, ~ wiai ~~ h are also of pl ~ tarmaecutïcal interest. Tea eluvxidation cm be affected throul; h trcatmctit with an appropriate reagent, for example rtaetcr-chloropcrbcnzoic acid (MC ', f'I3A). I'ïnal.ly, corrzpouncls of Clans 4 are accessed tltrough altcrttativc stazting matcrials alrcady containing a C3-keto functionality or stil?: ~ equcnt convcrsïoto of tltc C3-âzyclro ~ yl luta a kctone. Thc ketone can your be used ie> r constructïon "f 'c> thcr clcaired derivatives using standard methodologies known to those skillecl in flic art. Sintii ,, trly, compounds not spccilï caliy synthesized in the attached tloc ~; is oan bc. synthesired usïng l: nc ~ wn aml stancaard rrzcthoclologies from intermediates and proclucts clcscr-ihcd ihc ~ rcin.
l3io_ (o tical ~ ls ~ s Lüt ~ dit7, ~ l do lhe () ttctLotirt Rcc ~ cy ~ lor dis f'llc: allïnity ior tlze ouabain recchtor w ~ ls evaluated by the displaceznent of the specific j ~ il ~ cuabain binding ïn Madin-Daa ~ by caszinc kïdn ey (MI:) CK) cens. Tea specific binding to tüc rcccptc> r in dclïncd as thc difference bctwecn thc total binding and the nonspeciftc bïncling detern oined iv ihe presene, e of an excess of unlabeled ligand. Tea results are prescntecl as a percent of ~ control specific bïnding obtained in. the presence of varying oi concentrations ~ test compound. (T ablc. 1) Oua.bain was used as a positive control.
'hc iC ~ tt values (concentration rcvquirccl to rc; aclh 50% of maximum inhibition control ahe ~~ .ific binding) and Hill cc> cl ~ frc: icnts (nEi) were dctertmined by non-linear regression analysis of ihe compctition c; urvvs using lliil eequation cuz-ve fitting. Tea inhibition constants, K ;, werc calculateci frotta thc t: ~ 'lnerzg I'rusoff equation (K; =
ICSO / (1 + (L / Kr,)), v ~ ltcrc C, = concentration of radioligand: n the assay and K ~ ~ affinity of the radioligand for thc rcceptor.
'falïlc 1. Ouabain Receptor Bincling-Affinity for Repre.sentative Compounds of tea Invention __.__ ~ mpoitntl --...- lc ~ s ~ (lt ~) -_-. ~ '~ (~~~) ~ ._ ~ ._ ~ nti I - ".. .. ~.

t ~ t ~ 'lbai'1 t ~~ l __ ______ __ 1- ~
. ~ - ~ l __ ~.
_ -._._ ... _ ___. . _ ~
_ __. . _. 0.9 13a 1. ._ -______. . 7.1 I _-_ ___________ 7. () ~
___._ ________ ____._..___ _.___ ~. ~ ~. ~ 1.o 1 ~~ 1 __-_ _____ ~ _ ___ ___ ____ ____ _ ________ __;

_ 1? .Ct; 11.1) 1.1 1 (a l3inding curvcs fer these compouncis are presented in Figures Sa-Sc Irtlail ~ itimn nf Na +, h ~ - ~ 1'll'a.se Compounds were cvaluated for tl ~ eir ability to inhibit the aetivity of Na *, K + -ATPase lourificd fion u dag kidney rnoasurcd as l ~ crcent hydrolysis of A'fI 'in the presence and absence of the test compound. Inorganic hosphate release was determined according to ihc spectrophotometric literat.ttre rttethocl. ~~ Na +, Kt-A'rPase activity was c: alculated by 'Vls'OIaINlI. <1, read. Page 2G of 3l I) i ~~ ijali, s Ster ~ oiol tlrrcrla '~ = zre.v l'Wenl i2yplzccrtimt, lanzrary 2003 . Sul ~ tractint; thï: ouabain-insc.nsitivc phosphatasc activity froW the total phosphatase nct ~ Vity. In 'I'abl <; ?, t ho results tire pre ~ ented as a percent of cantrol values obtained in tlae prascncc c> f tlze test cornlaounds. C) uabain was agairt utïlized as a positive control.
'Cl u: It: S ~ vaincs (concentration c,: msin ~ j N / A''/ o inhibition of the control values) and full coef ~ ïcieuts wt: rc dctermined by taon-lincar regression analysis of the competition curves uaiy I Iill cc ~ uatiocl curvc littivg.
'I'alale 2. Nai, l ~'-ATI'ase In hibitory ll. ~ Ctivity of Representative Compounds of tlie Invention _ _ ._. _... _. _ ....____ ._ .. _ .___ _ ___ - ~ .__...._ w Compound ICso (wM) ~ night ; I _____________- () u'tbaitl --_ ____ __...__ ._ ._._.__. ___. ô..3 ~ _ ~ f _____ ... __ I. _. . _ _. _ __._ __ _ __ i ~ __- _ _ _..___ _. _ ~ ar 6 3 ~ ~ __ __ _.._ ~ .__.-___ _____. _.._ _____-_ -t) i ~ iC indicatcs less than:? 5 iô inhibition ut the highest tested concentration, 10 µM
finis inhibitory activity is prescntec.i grapi ~ ically in Figure 6.
CRRC ~ _clusio_n f'loarrnaccutical preparations utilrzing compounds of the invention can be applied therapcutic <illy or prophylactically tu the treatment of telly varions associated with the socli utn pump, including con ~; cstive huart failure, atrial fibrillation, cardiac arrhythmias and others. 'I'loe; compounds of the :; invcrytion can also be applied tu treat varions types of Hypertension including esscntial hypertension, thyroidism-induced hypertension and 1 ”-egnancy-associated hypcr-tension. In acidïtion, inltibitors of Na + .K + -ATPase also can be used as natriurctic / diurctic al; cnts. F ~ urther, the involvement of this enzyme / receptor ïn thc 'central nervous system (Cr ~ S) lzolds lite loossibility of tltese inhibitors being employed as ncuronto <lulators.
ndditionally, molccules of the inventüm have applications in cancer chemotherapy.
c) bscrvaüotzs of the positive cflccts of ihc cardiotonic stcroids on patïents suffering from cancer h ~ rve bceu known for somc lime.9 ° ~ »Some natural preparations Containing digitalis fatnily compounds have been sludicd in regard tu their antitumor effects.92 In . ~ dclitic "~, ouabain and rclateci tnoleculcs have becn reportcd tu have anti-apoptotic actiwities (W0 Ot) 17 ~ 700). Clnfor ~ utaateiy, the mechan.ïsn ~ of such activity down not yet heot ~ ciefined. 'hc involvemerrt of the endogenous digitalis compounds in tea laioeltemical pathways just now being revealed may be able tu explain and allow c ~ hpl ~> itation of these resultsys 1n; uidition tu the direct biological activity provided, t ~ luorinated steroids have utility in radiodiagnostic studios, in particular for imal = ing tumors with positron emitting tcmncrgraphy (I'L'fj. ~ 3 In thesc cases, the ~ sF radïoisolope must be employed.
advantages ~: ~ re tlnat this technidue allows ima ~; ing ol ~ cells. nerve tissue and tumors in roui Lime, ir7 1-7F (7.
. ~~ L ~~~ Il ~~ fi, ¿77e. Page 27 O ~ '31 1) i ~, Titc7li, s Steroid rl ~ 7crlo ~, eres l'crte7tt Afy ~ lieatic ~ n Janua7 ~ y 2003 ~ ZC '~~ C1 "CIICCS
1. Grcctc, I <., Ed., (.'Ctrdiac C¿lycosides, Sloringer-Verlag: Berlin, 1981.
Z. 5tcyn, ~ .5 .; 1-lccrden. 1 ~ '.IZ. Nut. Prcad. I (elr. 1998, 39 7.
3. Malcoltn, 5.13. I he L3ioclrerr7islr ~ of the C.'ardiac Glycosides, Chapman & Hall:
I, c> ndon, 1999.
4. Stnïth, '('. Digilalis G'lyco.s ~ iclos, Grune & Stratton: New York, 1985.
5. Ilcnoans, \. lon li-ctn.slcrcrtlion ht ~ thc .I \ ~ â ~ k '+ I'ttnzp: Influence of Membrane otc ~ nticrl and C,. 'ardictc (llycusideS {Acta Biotnedica Lovaniensa, No.
143), Coronet I3ooks: Philadclphia, 19 ') 7.
I ianson, JR Nczt. ~ r od. Izep. 1993. 10, 313.
7. lZuegg, UTL: yer ~ i ~ nticr 1992, 48, 1 102.
Por a rcview of Na ', K'-A7 ~ l'ase: l ~ aplan, JH Ann. IZev. Biochem. 2002, 71, 535.
~). Smith, rI'.WN I: n, t,> l. J. Mod. 1988. 31a, 358-365.
1 C). Revicw oF cndogenons cardinc glycosicics: Schoner, W. Eur. J. Biochem.
2002, 269, 2440-2445.
1 I. I_JS 5874423 describcs thc isolation of these- digitalis-like compounds from cataract lcns nuclei. The othcr salient fcaturc of these rnolecules is the presence of at tripeptide at thc 3-position. In addition the AI4.IS_unsaturated material is aussi rcportecl.
12. C ~ arg, R .; Gorlin, R .; Sntith, 't'.; Yosuf, SN Engl. J. Med. 1997, 336, 525-533.
13. Cioto, A .; Yatnada, K. 7i ° ends ire Pharm. Sci. 1998, 19, 201-a? 04.
14. Manunta, I '.; I-lamïlton, B.l .; Hahxlyn, JM Ilypcrtension 2001, 37 ~ part 2J, 472-~ 177.
1.5. Schciner-f3obis, G .; Scloncr, W. Nature Medicine 2001, 7, 1288-1289.
1 C3. Air, man, () .; Llhlén, l .; I ~ al, M .; Brismar, Il .; Aperia, A. Proc. Nat.
Acad. Sci. UST1 2001,> 8, 13420-13424.
17. Yackcr, M., l. C: Ardiol. the Izarraz. 1995, 26, S52-S56.
18. I'ackcr, M. .I; lrr7. ('o11. C'ctrdiol. 1993, ~? 2, I 19A-1'? (7A.
19. I ~ errebi-l3enrand, l .; Maixent, J.-M .; Clu-iste, G .; L clièvrc, LG
Biochirrz Biophys.
Actrr, 1990, 1021, 145-1 ~ 6.
20. C.'rambert, CJ .; Ilasler, 1J .; I3cggah, A.rI .: Yu, C .; Modyanov, NN;
Horisberger, J.-D .; helièvre, L .; Gecring, IC. .f Biol. Ç'hero. 2000, 275, 1976-1986 and references Clted thel'etIl, ? l. l3utt, AN; 'Tennant, BP; Gillingwater, SD; Shepherd, PS;
Swaminathan, R.
Ilyhertcns. h'es. 200 (), 23, S45-550.

22. Rcpke. KRI-1. Drzzg l ~ i, scovcyv I'oclav 1997, 2, 11C ~ -116.
Z3. Repkc, KRII .; Mcgges, R .: Weilaud, JJ Er ~~ yrne Iralrihïtion 1997, 12, 53-58.
24. Pedorova, OV; Bagrov, 1 '~. ~'. Anz. J. Ilypcr t erras ion 1_997, IU, 929-935.
25. Blanco, CT .; Koster, JC; Sanchez, Ci .; Mercer, R.Vf. l3ioclz ~ mistry 1995, 3 ~ 1, 9897-9903.
NI? () HlrtllA, Inc. Page 28 of 31 l>i,; itcllis ~ Steroid Artcalogtres atc ~ zt Al ~ pficrtliort Jartarary 200.

? G. Lichtstcin, D. in OToleculcrr and. ~ Ubcellular Car dialogy: Effect of Structure and h'rmuction, S. 5ideman and R. Beyar, I: ds., Plenum l'ress: New York, 1995, pp

23-30.
2'7. 13agrov, AY: Roukoyatkina, NL ~ Pinaev, AG; Dmitrieva, RL; Fedorova, OV
1: 'rrr ~. .l. Plrarmacology 1995,? Î'4, 151-158.
28. O'l3ricn, WJ; Lingrcl, J.I3 .; Wallick, h.'C. Arch. Biochem. Biophys.
1994, 310, 32-39.
29. 'Hon ~ as, R .; Gray, P .; Andrews, J. Advances in Drug Research 1990, 19, 311-562.
03. Replcc, KR1 (.; Sweadner, K..1 .; Weïland, J .; Megges, R .; Schon, R ..
Progress ire Or-rt, ~ T Re.wearclz 1996. 47, 9-S: Z.
31. Rcpke, KRI-i .; Wcilancl, J .; Menkc, K.-H. J: Enzyme Inhib. 1991, 5, 25-32.
32. Schünfcld, W .; Wcïlancl, J .; Lindig, C .; Masnyk, M .; Kabat, M.; M .; Kurek, AT.;
~ Viclia, J .; Ropke, K.IT.I ~. , Jrch. Phcrr-rraacol. 1.985, 3? 9, 414-426.
3; ~. Altnirante, N .; C; crri, A. Synlctt 19!) 8, i 234-1230.
34. C.'c: rri, A .; AOnïrantc, 1'v .; I-3arassi, I '.: Benicchio, A .; DeMunari, S .; Marazzi, G .;
Nlolinatz, L; Serra, F .; Melloni, P .., T Mecl. Claenz. 2002, 45, 189-207.
35. Scvillano, LG; Melero, C. I '.; Boya, M .; L.opez, JL; 'Tome', F .;
Caballero, E .;
C'arrCm, R .; Montero, MJ; Medarde, M .; San Feliciano, A. l3ioorg. Med.
Chem.
7999, 7, 2991-3001, ancl rcference ~~ cïtcd therein.
3C>. Mclcro, (~ .1 '.; Sevillanc ~, L. (- r .; CabaLlero, E .; rromé, F .; Carrbn, R .;
Montero, MJ;
San Fcliciano, A .; Medardc, M. 13rrorg. MeCl. Chern. Lett. 1998, 8, 3217-3222.
37. Quadri, h .; Bleached, G .; C, er-ri, A ... Fedrizzi, G .; Ferrari, P .; Gobbini, M .; Melloni, I '.; Sputore, S .; T'or ~ -i, hM. , J. Lied. C'herr. 1997, ~ 0, 1561-1564.
38. Fcnrandi, M .; Minotti, E .; Duzzi, L .; Molinari, L; Bleached, G .; Ferrari, P. .l.
(.'crroliuuusc. Pl7arnrcrcol. 20 () 2, 40, 881-889.
39. I- ~ 'crrari, l'.; Ferrandi, M .; 7, orielli, 1J .; Padoani, G .; Tripodi, G .;
Melloni, P .; Whitewashed <~. C.'ar ~ ditn ~ crscarlar Dru, t, T Rev. 1999, 1 ~ F, 39-57.
40. Ferrari, the .; rl'orielli, L .; Ferrandi, M .; adoani, G .; Duzzi, L .;
Florio, M .; Conti, F .;
Melloni, l .; Vescï, L .; Corsico, N .; E3ianchi, G. .l. the lZarrn. Exp. Ther.
1998, 285, 83-> ~ L.
41. Ferrari, P .; Ferrandi, M .; 'Frïpodi, G .; ~ i ~ oriellï, L .; Padoani, G .;
Minotti, E .; Melloni, I '.; I3ianchi, G. ~ T. the harrrr. Txp. Thcr-. 1999, 28b ', 1074-1083.
42. Gobbini, M .; Almirante, N .; Cerri.,, A .; Padoani, G .; Melloni, P.
Molec ° ules 1998, 3, 20-25.
43. ~ I'empleton, JF; Setiloane, l .; Kumar, VPS; Ya, Y .; Zeglam, TH;
LaBella, FS
.L Mcd. C.'herrz 1991, 3 ~ 1, 27 i 8-2782.
44. Tcrnpleton, .1.F .; Ling, Y .; Zeglam, 'I'.H .; Marat, K .; L ~ aBella, FS., ~
Cherry. Soc.
the erlc. Trans. 11992, 2503-2517.
45. rhempleton,: 1.F .; 1_, ing, Y .; ~ 'Eglam. TH; Marat, K .; L, aBclla, FS J
NLED. Chenr.
1993, 36, 42-45.
4C. Quadri, h .; Cerri, A .; Few, ~ ri, P .: Folpini, E .; Mabilia, M .; Melloni, P.
J. lt ~ lec ~.
C.'her -7. 1996, 39, 3385-3393.
NF (7KlMlA, Irrc, Pcrge 29 of 31 Di ~ italis Steroid fW crlogrtes l'Atertt ~ lhplicatiurT Jcrrruary 2003 47. 'I'empleton, JF; Ling, Y .; Marat, K .; LaBella, FSJ Med. This hem. 1997, 40, 1439-3 44G.
48. C: cn ~ i, A .; Altnirautc, N .; Barassi, P .; I3enicchio, A .; Fedrizzi, C .;
Santagostino, M .;
Schiavune, A .; Serra, 11 .; Zappavigna, MP; Melloni, PJ Metz 'Chem. 2000 2332 - '349?.
49. Smyth, DD; 'l'empleton, JF; Kumar, VPS; Yan, Y .; Widajewicz, W .;
LaBella, FS Ccn7. J. lrysiol. Pltar-Macol. 1992, 70, '723-727.
-_S0. Marient, JM; Bertrand, LI3 .; Lelievre, L, .G .; Fcnard, S. Arzneim.-Forsch. 1992 13 (11-1305.
51. 'l'cr.nplcton, JF; Kumar, VPS; I3osc, D .; Smyth, DD; Kim, RS;
LaBella, FS
(.'ccn. J. l'laysiol. Plaaf ~ nacrcol. 1988, b6, 1420-1424.
5'2. Wciland, J .; Schwabc, K .; llubler, D .; Schoenfeld, W .; Repke, KRHJ
Tn me ~~
hihi6_ 1987, 2, 31-36.
53. Saito, Y .; Kanemasa, Y .; Okada, Mr. Cltem. the harm. l3tall. 1970, 18, 629-631.
S ~ L. ~ I'cmplcton, JI ~ .; Kurnar, V.3'.5 .; Rose, D .; LaBella, FSJ Med.
Chem. 1989, 32, I 977-1981.
55. 'I ~ eypleton,, fl ~ .; Kumar, V.1'.5 .; Rose, D .; LaBella, FSJ Mecl.
Chem. 1989, 32, 1977-19 $ 1 and refcrences cited therein.
I6. Maixent, JM; Bertrand, LB .; Lelicvre, L, .G .; Fenard, S. Dru ~ l Res. 1992 ~ 2, 1301-1305.
7. Wcissenburgcr, J .; I-Ieckle, J .; Blour, M .; I'oiricr, JM; Jarreau, FX;
Jaillon, P .;
Cheymol, G. F2trtdarn- C.'lirz. the harn-tctcol. 1.989, 3, 67-78.
5s. llalpryn, B .; Fulton, R .; Fulton, V .; Panasevich, R .; Ogerau, T .; Fenard, S .;
l3aggioni, A .; Koenig, JJ; Chang, 1 '. the harnracologtsl 1987, 29, 135.
~ O. .larrcau, FX; lCoenig, JJ; f ~ enard, S. F: ur-. l7cart J. 1984, S
(suppl.F), 309-314.
t (). Bagrov, AY; Ro ~ ikoyatkina, Ivr.L; Pitaaev, AG; Dmitrieva, RL;
Fedorova, OV
lsur. .7. I'harrnczcolol;: y 1.995, ~ 7 ~ 1, 1'ïI-'158.
(~ l. l ~ edorova, OV; Bagrov, AY ~ lm., l. Il, vperterr, sion 1997, 10, x) 29-935.
f ~ 2. Welch, .1.T .; Eswarakrishnan, S. Flrrorirm in Bioorganic Chemistry, Wiley-Interscience: New York, 1991.
(i3. I ~ illcr, IZ .; Kobayashi_ Y .; Yagupolskïi, t, .M., Eds. ~ rganoflztorine C: 'ompounds ïrz ~ I ~ h ~ clicinal C.'hemist ~ y aracl l3ion ~ aedi ~. ~ Al ~ lppli, ~ atïoas ~, Elsevïer Science: Amsterdam, 1993.
t 4. Ojima, O .; McCarthy, JR; Welch, J.'T., E, ds. Bion2edical hronüers of Flztorit c ~
(: herfrishy (ACS Svtnposium Series No. 639), American Chemical Society:
Washington, I ~ C, 199C1.
(05. I ~ or a general rc, cent review on fluorine contaïning thcrapeutics in experimental and clinicat use: Ismail, FMDJ Fluonifze Clcenr. 2002. ~ '18, 27-33.
(> G. Rozen, S .; Ben-Shushan, G. lèt ~ ° ahedron l, ett. 1984. a '~, 1947-1948.
6 î. Rozcn, S .; Ben-Shushan, G. J C.) ri, T. Chem. 19811, 51, 3522-3527.
68. Rozcn, S .; Ben-Shushan, G. ~~ .layz. Itesort. Cltenr. 1985, 23, 116-118.
c ~~), l'-Iaas, A .; Kortmanu. DC'hetrr. ? ~ 3and. 1981, ll ~ l, 117f ~ -11 î9.
! 1 E (71i1 ~ 1 ~ 1A, I ~ tc, Page 30 of _31 r ~ i ~, = itcrlis Stcroicl Artcrlokues Pcttertt flphlir'aticrn Jantrary 2003 70. Hoff, DJ (7rg. C'herrt. 1970, 35,? _263, -2272.
71. Hanson, JR lVcrl, l'r ~ od. l2eh. 1998. l ~, '2c) 8.
72. Repke, KRI ~ I. Wcilancl, J .; Me: gges, R. ~ lr ~~, l ~. ~ U-. (::: lrcrrt. Jrzt.
.1d. lngl. 1095, 3 ~, 282.
73. Repke, KRII. Weiland, J .; Mcg ~; c ~, 1t. the R'0 ~ ,. , l ~ Ie (l. (:. 'hem. 1993, 30, 1 35-202.
74. Danie ~ vski, AR; Val enta, l .; Whüc, 1'.S. 7>'RRLL. oC. ticact. Sci.
C: lrerrz. 1984, 32, 29.
75. Stock, G .; Mook, R., Jr., I. ~ Lnr. (: 'ltcna. a ~ oc. 1983, Jl) 57 3',% 2 ().
76. Harnisch, W .; Morcra, 1 ~ .; Ortar, G. .l. t) ~ ry '. ('lrent. 1985, J0, 1990.
77. Daniewski, AR; Kabat, MI.M .; ~ Lasuyk; ~~ 1 .; Wicha, J .; Wojcicchowska, W .;
Duddeck, H. ..1. Gold ~ r. t: 'hen does. 1988, '3, 4 ~ SS.
78. Ruel, R .; Deslongchamps, I '. C: CTRT. .l. C ~ Does Herr. 1.992, 10, 1939.
79. Overman, LE; Rucker, 1'.V. Hetcrue: vc'lo.s ~ 20 () 0, ~~ ?, I2c) 7-1314.
80. Overman, I, .1 -:.; Ruckcr, 1'.V. I ~~ tr ~ r.rlrcc.lr ~ ort J, cett. 1998, 3 ~, 81. Hynes, J., Jr .; Overman, L, .I ~,; Tlas:, cr, '1'.; Ri.ickc.r, 1'.V. 'The etr-aherdron Lett. 1998, 39, 4647-4G S 0.
82. Deng, W .; Jensen, MS; Ovorman, I, .I ~;.; Ruckcr ~, I'.V .; Vionne; t, JP
.I. Ors. Chem.
1996, Gl, 6760-6761.
83. Chapdelaine, D .; T3elzilc, J .; Deslcmgc ~: hamps, I '. .I: () ry. ( 'Herrz.
2002, 67, 5669-5672.
84. Jung, MF; Pü-r_zi, G. (.lrg_ l.ett. 2003, 5, 13 7-140.
55. Jung, MF; Davidov, P. ~ lrt, c; ev ~. (.:. 'ircrr7. lrzt. .l ~; cl. 2002, 4 C, 4125-4128.
86. Stock, G .; West, h .; hec, 11.x '.; lsa, nc, 1: .C ~;.; Maunubc, S.. 7. .1rrr.
(.. 'lterra. .Soc. 1990, 1 J ~. 10660-10661.
87. P. Deslongchamps Pure ~~ AJ ~ J ~ I. (',' llern 1992, 6. ~, 1831.
88. Cohen-L, Lüla, R .; R.imon, G .; Morau, A., Bina. .T. the lty. ~. ~ iul. 1993 264, I161-F65.
89. Fiske, (~ .H .; SubbaIRow, 1 '., L. L3iol. T.'lueur. 1925, GC, 375-40 ().
s ~ 0. Stenkvist, B. C> ncol. 7ZeJ ~. 199!), 3, 4 ~) 3 .. ~ lcJp.
~.> I. Haux, J. aLTeclical Ivypotltca ~ e, s ~ 1999,. ~ 3, '~ 43_54 ~.
Smith, J.f'1 .; Madden, '1- .; Vi, ljeswarapu, M .; I \ 'ewman, R..f1- I3iochont.
Pharrnacol.
2001, G2, 469-472.
~) 3. ICatzenellenbogen, .1.A. , .l T'h.rorirre ~ _'ITenz. 2007, Jt) 9, 4-9-54.
NEOKIA.IIA, hrc. Page 31 of 31 Digitalzs S'ter ~ oid ~ lrzalogzre.s Patent ~ llylic: atlcm Jarzzrary 2003 Brute formula ; CzZH3; O ~ F.
1 H NMR (300 MHz, CL) C.1;). 8 (, ppm): 6. ~) 1 (1 l I. dd, .I = 15.6 Hz and J =
8.7 Hz, CH = CHCOZMe), 5.83 (1 H, J = I ~> .6 11z, (: Fl = Cf _ ((_ '. OzMe), 4.14 (1H, t, J =
2.6 Hz, RR'C140H}, 3.73 (311, s, C: C) ~ C; H3.),: '. ~~ 1 (II l, cl, J --- v 8.7 Hz, RR'GI ~ CIi = CHCO2Me), 2.22-1.03 (23H, m), 1.00 (3H, d, J _ ::. '.I. I 1-l; r, Cl l ~:.
RMN ~ 'C (75 MLIz, C', DC1;), c3 (ppnu): 14'x.70, 149.57, 121.97, 66.93, 51.45, 51.33, 49.00, 45.09., 44.81, 41.27, 34.39, 34.27, 3> .26,: vl .0t1. 3041, 30.f) 6, 29.96, 29.37, 26.97. 25.04,

24.69, 20.98, 20.74, 16.81, 16.74 (3 more peaks).
1R (CL1C13), v (cm- ~): 3501. '> 93 ~', 2875, 1718, 1654; 1540, 1558, 1437, 1281, 1216, 1002.
SM (m / e) (Tp ° of the source: 100 "C): s64 M, 344 (MI-1 h) ~ '.
SMHR (Tp ° from source: i 00 ° C) 'calculated for C ~~ 113_ ~ C) ~ F
(M) +: 364.2414, found 364.2403 ~ 0.0011.
Ester a, ~ 3-unsaturated 152b VS
Me CN ME CHO Me H ~~ __-_ Y E-1 ~ f '~ ._..___ ~ ",. H
F ~ \ H. ~ F ~ \ ~ HF
HO HO HO
142b 153b 152b 02Me Ä a solution of nitrile 142b (4 m ~ ;, 0.013 mmol) in toluene (1 mL) â -60 ° C has been added a 1.5 M düsobutyIaluminiurn hydride solution in toluene (26 pL, 0.039 mrnol) and the mixture was stirred at -60 ° C for 1 h. I) u sodium sulfate decahydrate has been added and the mixture was stirred at room temperature for 1 h. The precipitate has been filtered on a thin layer of salt and the. f Itrat was evaporated to give 1 "crude aldehyde 153b (4 rng).
. ~ a suspension of sodium hydride 60% in oil (5 mg, 0.13 mmol) with temperature ambient in tetrahydrofuran (i mL) was added phosphonoacetate trimethyl (2l uL_ O.13 mmol). The mixture was stirred at room temperature for 15 min and the crude aldehyde 153b (4 mg, 0.013 mmol) was added by cannula. The resulting mixture has been stirred at room temperature: s0 cnin and mre aqueous scalution of ammonium chloride saturated has been added. The aque phase ~ .rse ar Was extracted ~ c: with d ~ .r dichloromethane and phases organic together were. dried with c: magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (4U ~% ethyl acetate, 60% hexane) to give the ester cz, ~ -unsaturated 152b under ti ~ rmo of a colorless oil in a 9: 1 mix (~ 'j: a) inseparable (3 mg, GO ° ~ o for ~' stages).
Raw formula: C :, ZH33031 :.
1 H-NMR (300 ml ~ z, CDC1;), cS (ppm): 7.07 (1l-1. Ddd, .I - 15.5 1-Iz, J = 10.7 Hz and J = 1.8 Hz, C'H = CHCOZMe), 5.65 (1H, J = - 1 ~ .5 f-1. ~, CI1 = C '. (W1C', OzMe), 4.i4 (1H, t, J =
2.7 Hz, RR'CHOH), 3.71 (3I-1, s, COZCI t3), 2 38 (1H, td, .1 - 8.5 Hz and J = 3.2 Hz, RR'CHCH-Cl-IC'.O ~ Me), 2.17-1.03 (î? 3H, n ~), 0.x) 0 (3-H, s, CHI).
R.MN '~ C: (75 MHz, (: DC13), 8 I ~ ppm): 15 ~~ .55 "153.49, 1' I 9.75, 66.93, 53.93, '~ 1.35, 45.09, 44.81. 41.43, 38.08; 37.98, 34.18, 3s. ~ '7, .t1.0 ~), 30.75, 29.37, 26.97, 26.66, 25.42, 21.09, 21.01. 15.63. 15.55 (1 more peaks).
IR (CHCl3), v (cm ~ r): 3501, 2931, 2865, 17! 8, 1654, 1541, 1458, 1437, 1269, 1221, 1000.
SM (m / e) (Tp ° from source: lof ° C): 364 M ', 344 (MI-lr)'.
SMHR (Tp ° from the source: 100 ° C); calculated for C ~ 21 ~ 3303F (M) +
: 364.241.4, found 364.2403 ~ 0.0011.
Fluorinated compound 1.57 ~ O ~
.O-, "t.7 'O
Me J ~ 'M
Met '~'~> -_ _.... Me ~~
i ~ H ~ / OH HF
agio ~ wa ~ o 154,157 ~ a solution of the trifluoride of (dïëChylamino) sullùre (6 ip, I_ ,, 0.46 mrnol) in the dichloromethane (10 mL,) at 0''C 'were added pyridinc (1 n ~ I.), a acid solution hydrofluoric: pyridine (1 ml,) and alcohol 144 (98 mg, 0.23 rnmol), l ~ e mixture was stirred at room temperature ~~ te during l. '> lu and an aqueous solution; use sodium bicarbonate saturated was added quietly. 1, a pha ~~ e ~ aclueus ~~~ has been extracted with dichloramethane and the combined organic phases were sècl ~ é ~ s avc; c sulphate magnesium, filtered and evaporated. the residue obtained <z was purified by flash cloromatography (50%
ethyl acetate.
50% he ~ ane) to give the compound fluorE ~ xS7 in the form of a white solid (18 mg, 18%).
Raw formula: C25H3sO4F.
YF: 173-174 ° C :.
RMN 11-1 (300 MHz, CDC ", Is), ~> (pprn): 5.8Ci (1H, s, IZR'C: == CI-1), 5.08 (1H, t, J = 2.5 Hz, C'NOAc), 4.87 (1H, d, J - 18. (> Hz, R-t'.HIiO ~ <. 'R'), 4.78 (11-1, dd, J - 18.0 and J = 1.7 Hz, RCH ~ iU2CR '), 2.85 (1H, dd, J == 9.1 Hz and: I - 4.7 l ~ Iz, RR'CHC (R ") = CH), 2.30-2.07 (3H, na), 2.06 (3H, s, CH ~ CO ~ R), 1.98-1.06 (I 8H. nr), 0.99 (31-l, s, CH3), 0.92 (3H, s, CHI).
RMï1 ~ 'C (76 MHz, C.'. L>C1;), ~ ï (ppm): 173 21, 170.66, 1 17.84, 1 1 1.42, 72.99, 72.84, 70.17.
50.59 ,, 39.31, 39.21, 39.03, 38.75, 36.'2, _ ~ 6.1 = 1. 36.01, 35.08, 31.48, 31.16, 30.40, 26.47,

25.88, 24.99, 23.64, 21.47, 21.06, 20.?2, 1 ~. s3. 1 sec. 45 (3 more peaks).
NMR ~ 9F (300 ml-Iz, C; I) C13), ci i_pprn): -73. i ~) to -73.60 (1F, m, RIZ'R "C ~.
1R (C1-IC1 ~), v (cm- '): 3020, 2944, 1785. 17-17, 1624, 1449, 1 379, 1261, 1215, 1026.
5M (m / e): 436 (MNI-In) +, 419 (MH) ~ +.
SiVII-IR: calculated for C ~~ H ~ H (~ al ~ (Ml l) ~: 419.2> 97, l. Found: 419.2607 ~
0.0013.
cz] v == + 12.7 ° (c 1.2, C. '. l-IZC: 12).

14 / ~ Fluorodigitoxigeninc 7 O
H

, ~ 1 a solution of acetate 1> 7 (12 mg, 0.028 mmol> in methanol (1 mL) has been added 5 drops of concentrated hydrochloric acid and the mixture was; restless ambient temperature All night long. (.Jne aqueous bicart solution ~ saturated sodium onate was added and phase aqueous was extracted with dichlcorontéthtmc. 1, organic phases reunions were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (60% ethyl acetate, 40% hexane) to give the 14 ~ -Iuorodigitoxigenin 7 in the form of a solid tin (') mg, 82%).
Gross formula: CZ; I-13 ~ 03F '.
17.F. : 203-205 ° C.
RMN I1-1 (300 MHz. CL.) C13), ~ (ppn7): 5.8fï (11-l, s., RR'C = CII), 4.87 (1H, d,, T - 18.0 Hz, 1ZCHHO ~ CR '), 4.77 (II- ~, dd, J == 18.0 and J - 1.7 Hz, RCHH02CR'), 4.13 (1H, t, J = 2.7 Hz, t ~ 'HOH;), 2.84 (1H, dd, J = 9.2 Hz and .1 = -: L.6 11z, RR'C1 ~ IC (R ") --- CI4), 2.29-1.20 (22H, m), 0.98 (3H, s, C: ~), 0.92 (3H, s, (; H).
3 C NMR (75 MHz, CL) C13), cS (ppm): 17.x, 24, 117.84, 111.51. 72.99, 72.84, 66.73, 50.64, 49.66. 49.40, 39.39, 39.29, 39.04, 38.76, 35.99, 35.90, 35.33, 33.30, 31.48, 31.15, 29.50, 2'1.86, 26.48, 26. (.) 2. 23.69, 2i.1 $, 20.74., 1 ~. ~ 5, '15 .47 (3 more peaks).
1R (CF-1C1;), v (cni '): 3685, 3020, 2943, 1747, 1624, 1522, 1424, 1215, 1032.
~ M (tn / e): 37 (~ M +. 358 (M-Hz (~) +.
SMHR: calculated for C25H_ ~ O_, F (M) ': 376.2414, found: 376.2425 ~ 0.0011.
a] n = - ~ 6.0 ° (c 0.7. CHzCl2).

~ 04 Is'.ayon-X: diffusion of pentane in ethyl acetate.
~ (: 'fluorinated omposed 160 , Y. ”O

Ac0 ~ --O? - AcC) - --C
OAc. 3 OAc r '~ a solution of the trifluorurc of (diethylannino) sulfide (59 pI ~, 0.46 mmol) in the dichloramethane (10 mL,) at 0 "C was added pyridine (1 mL), a solution acid hydrofluoric acid: pyridine (1 mL) and alcohol 159 (140 mg, 0.15 mmol}. 1.e mixture was stirred 4t the ambient temperature during I.5 he ~ an aqueous solution of sodium bicarbonate saturated was added quietly. The aqueous phase was extracted with dichloromethane and the combined organic phases were dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (100 ° / ~ ether) for give the fluorinated compound 160 under the form of a solid: white (45 mg, 32%).
the raw formula: C, ~ yH ~ 20u1 ~.
RMIV 'H (300 MHz, CDC13}, c ~ (ppm): s.8 ~ (1H, s, RR'C = CHa, 5.43-5.33 (4H, m, ((: I = IOAc) 4), 4.89-4.67 (51-I, m, ECCE-I, òO ~ CR "and (ROCHOR ') 3), 4.51 (1H, dd, J = 9.9 and J = 2.9 I-Iz), 4.00 (111, s}, 3.91-3.76 (3H, na. (CI-J ~ (R) C''1_lOR ') 3), 3.29 (2H, dt, J = 9.8 Hz and J = 3.1 Hz), 2.83 (11-1, dd, J == 9.0 11z and J - 4.7 ltz, RR'C14C (R ") = ClJ), 2.100 (3H, s, CH C02R), , .097 (3H, s, C: ly ~ 3CO ~ R), 2 .095 (3I-i, s. C>rl; C () zR), 1.99 (3H, s, CH; C () ~ R), 2.2 5-1.17 (36H, rn), 0.94 (3H, s, CH}. 0.91 (3H, s, C: ~).
RMT''C (7 ~ MHz, CDCl3), b (ppm): 173.27. 170.23, 169.94, 117.78, 109.17, 98.83, 98.72, ~~ 5.82, 79.55, 79.51, 72.99, 72.87, 72.30, 69.07, 69.61, 68.99, 68.92, 67.83, 67.35, 50.61.

39.39, 39.04, 38.76, 36.26, 36.18, ~~ 6.12, ~~ .84, 35.72, 35.09, 31.46, 31.13, 30.08, 29.67,

26.62, 26.49, 26.17, 23.57, 21.36,.? I .30, ~ '1. i 9, 20.96, 20. 72, 18.18, 17.97, 14.62, 15.52, ! â.44, 15.25.
1R (Cf: -1C13), v (cm- '): 2938, 2877, I i80, 1747.! 371, 1245, 1167, I 155, 1095. 1057, 1025.
a] D ~ - +. 52.8 ° (c 1. l, CI-.I? Cl,).
(-. 'omégé fluorine 163 O. \ = U
M e / ~
'....__...
~~ '' HO ~ DD ~ _>
CHap ~ \ ~ ~, - CH30 , ~ -O ~ '162 ~ ~~ G, ~' 163 GP -OÖGP 'GP = CI3CCH, OC (O ~ GP ~ -D ~ GP GP _ = CI3CCH20C (O) i3 3 To a solution of (diy-thyfamino) sulfide trifluoride (63 ~ .L, 0.48 mmol) in the dichloromethane (10 ml,) c ~: 0 ° C. has been added pyridine (l ml.), an acid solution hydrofluoric: pyridine (1 ml.) and alcohol 162 (228 mg, O.16 mmol). The mixture was stirred at room temperature for 1 h and an aqueous bicarbonate solution sodium saturated was added quietly. the aqueous phase was extracted with dichloromethane and the combined organic phases were dried with sulphate magnesium, filtered and evaporated. The residue was purified by flash chromatography (80%
ether, 20%
hexane) to give the compound flut ~ rc; 16: 3 bags form a white solid (65 mg, 29%).
Crude formula: C ~ 3H6sOz, Cl ~~ l ~ '.
R ~, ~ I1 ~ I 'H (300 MHz, C'DC.I;), cS (ppm;: 5.8â (1I-1, s, RR'C-CH), 5.46-5.43 (2H, m, (CHOCOZCHZCCI;) 2), 5.34-5.32 (2H, rn, (C: F1OCOZCHZCC13) Z), 4.93-4.68 (13H, m, RCH O ~ CR 'and (ROCHOR') 3 and (C13CH CI ~ CK '), ~), 4.44 (1 H, dd, J = 9.9 and J =
3.0 Hz), 4.03-4.01 (1H, m), 3.92-3.84 (31-I, r ~~, (C.'H; (R) t: JHCjR ') 3, ~. 3.29 (2H, dt, J =
9.4 Hz and J = 3.1 Hz), 2.83 (III, dd, J = 9.0 T-lz and J == 4.5 1-1r. RR '(' 1 ~ IC '(IR ") = - CII), 2.24-1.14 (36H, m), 0.94 (3H, s, C11;), 17.91 (311, s, CI13).
RMN ~ 3C (75 MI-lz, C: DC'13), ~ 'i (pp ~ n): 1 "4.1 ~ I_ 173.30, 153.6' ?, 153.30, 153.24, 153.06, I 17.79, 1 II. S 1, 109.15, 98.6 (1, 95.28, 94.70. 94. ~ 7, 94.19, 94.04, 79.27, 79.22, 76.82, 76.67, 76.60, 75.70, 75.x.2, 73.00, 72.04, 72.86, ï2.14, 68.73, 68.68, 67.32, 50.63, 49.65, 49.40, 39.39, 39.30, 39. () 4, 38.76, x6.23, 3f> .15,: x: 5.65, 35.54, 35.10, 31.57, 31.47, 31.15, 30.11, 29.95, 26.59, 2 (i.49, 26.16, 23.60. 21.21, 20.72, 18.07. 18.02, 17.69, 15.54, 15.46, 14.12.
1 R (CI-tCl3). v (cm- '): 2937. 176 (1, 1449, 138 i, 1252, 1 J 70, 1054, IO
I 7.
(a] n = _ + 42.7 ° (c 1. I, C'I-IZC'I ~).
14 / j-Fluorodigitoxin 161 ~ O ~.: O
Me Me ~ ~. _%
~. ~~ 1H, ~ F __. _ -____, CH ~~ '~ CH
sO sO
1 ss ~ -o ~ ~ s1 GP O iJ GP = Cl3CCH20C (O) H 0 ~~~
OGP ~ OH .3 r1 a solution of compound 163 (65 mg, () .044 mmol) in acetic acid (1 mL) was added a small zinc spatula and the mixture was stirred at temperature ambient all the night. 1, the precipitate was filtered with dichlorornethane and methanol and the organic phase a been washed with an aqueous saturated sodium bicarbonate solution. 1, a organic phase a was dried with magnesium sulfate, filtered and evaporated. The residue obtained was purified by flash chromatography (10% methanol, 90% dichloromethane) to give the 14 ~ 3-i: luorodigitoxin 161 in the form of a white solid urn (20 mg, Ei 3%).

Fc> gross formula: C '.. ~ 1N ~ 30i? 1 ~.
1 H NMR (300 MHz. CI) C, 13). b (ppm): x.85 (1H, s, fZR'C ~ = CH), 4.93-4.74 (5H, m, RC: H ~ O ~ C: E2 'and (1ZOC1-IOR') ty), 4.26-4. '? 3 ('? Il, na, C1, I011), 4.14-4.11 (1H, s, CHOH), 4.02 (1H, s, CE ~ Ol-1), 3.87-3.72 (3f-I, 1n, (t: ~ 'H-L3 (lyC'.1-IOR');), 3.32, -3.26 (11-1, m), 3.22 (2H, dt, J =
9.7 Hz and J '- 3.0 1-Ez),: 3.04-'2.97 (2H, 3n), 2.83 (1H, dd, J = 9.1 Hz and J
4.6 I-lz, RR'CHC (R ") - = C1-1), 2.37-1.18 (381-1, m), 0. ~~ 4 (= X13, s, Cl'I;), 0.90 (3H. S, CH3).
RMN 1jC (75 M1-1r, CDCI ~), ti (ppan): 174.33, 173.55, 117.72, 111.60, 109.24, 98.24, 98.16, ~> 5.34, 82.51, 82.13. 73.00, 72.94, i2.64, 7: 2.47, 69.51, 68.24, 68.03, 66.40, 66.31, 50.65, 49.67, 49.42. 39.41,: 39.31, 39.04, 38.76, 3'7.78, 37.10, 3 (i.66, 36.23, 36.16, 35.09, 31.47, : $ 1.15, 30.16. 29.73, 26.62, 2 (i.50, 26.09, 23.57, 21.22 "20.72, 18. I 5, 15.55, 15.47.
1R (C'HC13), v (ctn ~ '): 3446, 21> 34, 1782, ~ 744, 1654, 163ti, 1449, 1381, 1319, 1218, 1164, : I 29, 1066, 1 O13.
[~ x] ~~ _ ~ -X8.3 ° (c 0.7, Cl-l ~ Clz).
Ester a, ~ -unsaturated 8 Me CHG Me CHO
___. ,. Me ~ '~' ~ _ ~ _ ~. Me , - ~ w, .. /
~~ 'HFHF
HO ~ -, H ~ / ~
7,164 8 , Ä a solution of 14 ~ -fluorodigitoxigenin 7 (18 mg, 0.048 mmol) in acetone (3 mL) were added ruthenium oxide (: mri, 0. () 048 mmol) and the periodate sodium (26 mg, 0.12 mmol) dissolved in water. The mixture was stirred at temperature ambient for 30 min and the same amount of ruthenium oxide and sodium periodate were added. The mixture was stirred 1 ~ min aupplément4iires then isopropanol was added (1 mL) and stirring was continued for 15 min. The precipitate was filtered and rinsed at acetone and filtrate was concentrated; to remove '''acetone. 1, the residue obtained has been dissolved in methanol (I mL) and sodium borol hydride ~ 4 mg, 0.096 mmol) was added. The mixture was a ~ ïté u the ambient tcmpératurc for ~ + 5 mira then acidified with a acid solution hydrochloric 1 N up to a pli do ri. L, el ~ çric, sodium date has been added to this solution then the obtent.ca mixture was stirred at ~ room temperature for 3 h. The rushed was filtered and rinsed with water, 1.a phase aquer ~ has been extracted with ethyl acetate and phase, organic materials were dried with magnesium sulfate, filtered and evaporated pc> ur give aldehyde 164 comesporvdant (1'S ny.l.
t1 a suspension of sodium hydride 60 ~ rt> dau ~ s oil (I9 mg, 0.47 mmol) was added on trimethyl ploosphonoacetate (76 l ~ L ,, 0.47 mmol) and the mixture was stirred at temperature room for 15 min. The crude aldehyde I64 (15 mg) was added by cannula and the The resulting solution was stirred for 3 ~ 1 min at room temperature.
1_1ne solution of saturated ammonium chloride was added and the aqueous phase was extracted with dL acetate; ethyl. I, the combined organic phases were dried with sulfate magnesium, filtered and evaporated. The residue obtained was purified by-flash chromatography (5 (>%> ethyl acetate, 50% hexanc) to give the "13-unsaturated ester 8 one form colorless oil (5 mg, 29%, 4 steps).
raw formula: Cz ~ H3,03F.
IsMM ~ H (300 Mllr, CDCI ~), b (pprn): 7. () I (I II, ddd, J = - 15.5 I-Iz, J =
10.7 1-Iz and J = 1.9 I ~ z, CH = CHCOZMe), 5.64 (11-l,: l _ = 15.5 Hlz, CH -CH ('OZMe), 3.73-3.60 (1H, m, RR'CHOH) 7.71 (3I-I, s, CO ~ C >> I3), 2.38 (llri, td, J = 8.2 1-lr and J = 3.1 Hz, RR'CHCH = CHC; OZMe), 2.18-0.95 (22H, m), 0.94 (3H, s, CH1_ ~), 0.88 (3H, d, J = 0.9 Hz, C: H3).
1MN ~~ Cï (75 MI-1-r_, C.DC1 ~), 8 (ppm): 167.14, 153.51, I 5 i.46, 119.75, 111.64, 71.61, 53.95, 51.36, 41.58, 39.28, 39.00, 38.08, 37.97- 36.71, 36.60, 36.25, 35.03, 34.75, 31.11, 30.78, 30.56, 26.63, 26.56, 23.24, 21.26, 20.26, 15.62, 15.53 (4 more peaks).
1R (CHCl3); v (cm- '): 3380, 2940, 28ti6, 1723. 1651, 1452, 1265. 1225, 1162, 1079, 1038.
~ M (mi'e): 378 Yl ~, 358 (M-HF) +.
SMHR: calculated for C ~ 3H35G3F '(M ~~': _ '78 .2570, found. 378.2562 ~
0.0011.
[cz] ”= _ + 25.8 ° (c 0.4, CH ~ CI ~).