Synthesis of vinyl pyrophosphonate analogues of farnesyl pyrophosphate: New potential inhibitors of farnesyl protein transferase

Article abstract of DOI:10.1002/hc.10081

The synthesis of new bioisosteric analogues of farnesyl pyrophosphate where a vinyl pyrophosphonate replaced the pyrophosphate moiety is described. These compounds have been prepared using a Horner-Wadsworth-Emmons procedure and a modified Michelson react

Full text of DOI:10.1002/hc.10081

Heteroatom Chemistry
Volume 13, Number 7, 2002
Synthesis of Vinyl Pyrophosphonate
Analogues of Farnesyl Pyrophosphate:
New Potential Inhibitors of Farnesyl
Protein Transferase
Ibrahim Zgani, Chantal Menut, and Jean Louis Monte´ro
Laboratoire de Chimie Biomole´culaire—UMR 5032, Universite´ Montpellier II, ENSCM,
8 rue de l’Ecole Normale, F-34296 Montpellier Cedex 05, France
Received 21 February 2002; revised 24 April 2002
that remains locked in an activated GTP-bound state
ABSTRACT: The synthesis of new bioisosteric ana-
thereby relaying an uncontrolled proliferative signal
logues of farnesyl pyrophosphate where a vinyl py-
resulting in malignant transformations [3,4].
rophosphonate replaced the pyrophosphate moiety is
To function in the signal transduction, Ras pro-
described. These compounds have been prepared us-
tein must be bound to the cell membrane. This bind-
ing a Horner–Wadsworth–Emmons procedure and a
ing is facilitated by a sequence of postranslational
modified Michelson reaction. They have been eval-
modifications [5]. It has been shown, in particu-
uated for the inhibition of farnesyl protein trans-
lar, that farnesylation of Ras is necessary for proper
ꢀ
C
ferase. 2002 Wiley Periodicals, Inc. Heteroatom Chem
functioning in cell signaling; this prenylation is cat-
alyzed by an enzyme, protein farnesyl transferase,
and this discovery provides the rational of the devel-
13:654–661, 2002; Published online in Wiley InterScience
(www.interscience.wiley.com). DOI 10.1002/hc.10081
opment of FTase inhibitors as a new type of antipro-
liferative agents.
INTRODUCTION
Several types of farnesyl transferase inhibitors,
especially peptides or peptidomimetics, have been
designed for use as anticancer drugs depending on
their mechanism of activity [6]. Farnesyl pyrophos-
phate analogues as potential chemotherapeutic
agents have also been extensively studied. Among
them, several mono-, bi-, or triphosphonate com-
pounds, such as compound 1a, have been synthe-
sized for an examination of their ability to inhibit
FTase [6,7]. Furthermore, to gain information about
the interaction between proteins and prenylated
pyrophosphate, compounds that incorporate a ben-
zophenone crosslinking group have been synthe-
sized to exploit their photochemical properties. In
this respect, the synthesis of compounds 1b and 1c
that contain stable ether linkages between the pho-
toactive benzophenone and the isoprenic moieties
has been described. Both compounds were found to
Protein prenylation is an important posttransla-
tional protein modification, which appears to play
a major role in protein association with membranes
as well as in protein–protein interactions [1]. This
process involves covalent addition of either farnesyl
or geranyl–geranyl groups to cystein residues local-
ized at or near protein’s C- terminus.
Ras proteins play an important role in a signal-
ing the pathway that controls cell proliferation, e.g.
mutations in Ras proteins have been estimated to
be intimately associated with 30% of all human can-
cers [2]. Ras gene mutations produce mutated Ras
Correspondence to: Chantal Menut; email: cmenut@univ-
montp2.fr.
c
ꢀ
2002 Wiley Periodicals, Inc.
654

Synthesis of Vinyl Pyrophosphonate Analogues of Farnesyl Pyrophosphate 655
be competitive inhibitors of yeast protein farnesyl
transferase, with respect to farnesyl pyrophosphate
(FPP), the presence of benzoyl benzyl group induc-
ing a higher affinity for FTase when compared to FPP
[8,9].
Here, we report the synthesis of bioisosteric
analogues of isoprenoid pyrophosphates: vinylpy-
rophosphonate derivatives, which are known to be
stable towards enzymatic hydrolysis.
The first compound targeted was the vinyl py-
rophosphonate 2a corresponding to phosphonate 1a
whose inhibitory activity on FTase had already been
observed [6]. Compared to 1a, pyrophosphonate 2a,
which contains one more phosphorus atom, is more
bioisosteric with respect to farnesyl pyrophosphate,
and therefore an increased activity on FTase may be
expected.
Other components are vinylpyrophosphonate
analogues of 1b and 1c, which were synthesized to
test the effect of the replacement of the pyrophos-
phate moiety on their inhibitory activity on FTase
(Scheme 1).
SYNTHESIS
Compounds 2b,c and the triphosphorylated com-
pounds 3b,c were synthesized in a nine steps
sequence as illustrated in Schemes 2 and 3.
Dimethylallyl alcohol (4b) [8] and geraniol (4c)
were first protected as THP derivatives by re-
action with 3,4-dihydropyran and pyridinium p-
toluenesulfonate (PPTS) [10]. The resulting ethers
5b and 5c were oxidized with t-butylhydroperoxide
and catalytic H2SeO3 in dichloromethane afford-
ing 6b and 6c (38 and 47% yields respectively)
[11], which were then treated with NaH and 4-
(bromomethyl)benzophenone to give 7b and 7c (36
and 49% yields) [8]. Deprotection of compounds 7b
and 7c was accomplished with catalytic PPTS in
methanol and afforded 8b and 8c in quantitative
yield [10]. The alcohols 8b and 8c were oxidized with
pyridinium dichromate in DMF and yielded the cor-
responding aldehydes 9b and 9c in 68 and 70% yield,
respectively [12]. This reaction is very clean, and the
isolation of products is simple. There is no overoxi-
dation of the aldehydes and no E to Z isomerization
of the aldehyde ꢀ–ꢁ double bond such that we ob-
served with manganese dioxide [13] or the Swern
reagent [14]. Farnesal (4a) was obtained according
to the same procedure (91%) from farnesol.
Treatment of the aldehydes 4a, 9b, and 9c with
tetraethylmethylene diphosphonate and NaH in ben-
zene gave via a Horner–Wadsworth–Emmons con-
densation the expected vinyl phosphonates 5a, 10b,
and 10c (60, 95, and 75% yields) [15]. The resulting
SCHEME 1 Inhibitors of FTase (1a–c) and their pyropho-
sphonate analogues (2a–c).

656 Zgani, Menut, and Monte´ro
SCHEME 2 Synthesis procedure of aldehyde precursors.
phosphonates were converted to the corresponding
phosphonic acids 6a, 11b, and 11c by standard
treatment with bromotrimethylsilane and pyridine
in CH2Cl2 (84, 70, and 85% yields, respectively) [16].
The synthesis of vinyl pyrophosphonates was
performed using modified Michelson’s conditions
(detailed in the experimental section) [17]. Com-
pounds 6a, 11b, and 11c were first activated by
SCHEME 3 Synthesis procedure of pyrophosphonates from aldehydes.

Synthesis of Vinyl Pyrophosphonate Analogues of Farnesyl Pyrophosphate 657
the reaction of their tri-n-butylammonium salt in
spectra were measured with a Jeol DX-300 spectrom-
eter in the FAB+ (NBA) or FAB− (GT) ion mode.
dry THF with diphenyl phosphorochloridate and
tri-n-butylamine. Compounds 7a, 12b, and 12c
obtained by this reaction were then combined
without any purification in dry pyridine with the
tri-n-butylammonium orthophosphate to give com-
pounds 2a, 2b, and 2c in 49, 48, and 57% yields,
respectively, after purification. Besides, formation of
the corresponding triphosphorylated derivatives 3a,
3b, and 3c was observed and these compounds were
obtained in 18, 6, and 17% yields, respectively. The
formation of compounds 3a, 3b, and 3c is due to
the fact that the activation is not complete, so that
the remaining diphenyl phosphochloridate can react
with the tri-n-butylammonium orthophosphate pro-
General Procedure for the Preparation
of the Aldehydes from Corresponding Alcohols
To a solution of pyridinium dichromate (4.45 mmol,
1.2 eqiv.) in DMF (30 ml), which was cooled to −30◦C
and maintained under nitrogen, the alcohol 8b, 8c,
or farnesol (3.56 mmol, 1 eqiv.) was added and the
mixture was stirred for 3 h, then allowed to warm to
room temperature; finally water (100 ml) was added.
The aqueous solution was extracted with ether (3 ×
50 ml) and the combined ether extracts were washed
with aqueous HCl (1 N, 80 ml) then with water, dried
over anhydrous Na2SO4, and concentrated in vacuo.
The oily residue was chromatographed on a silica
gel column with ethyl ether – petroleum ether (3:7)
as eluent.
1
ducing the P-diphenyl pyrophosphate. The subse-
quent displacement of the diphenyl phosphate group
by the tri-n-butylammonium salts of compound 2a,
2b, or 2c conduced finally to compound 3a, 3b,
or 3c.
BIOLOGICAL EVALUATION
(E)-3-Methyl-4-(4ꢁ -benzoylbenzyloxy)-2-butenal
(9b). (712 mg, 68%).1H NMR: δ 2.15 (d, J5-2 = 1.4
Hz, 3H, H5), 4.1 (s, 2H, H4), 4.62 (s, 2H, OCH2C6H4),
6.21 (qd, J2-1 = 8.0 Hz, 1H, H2), 7.42–7.55 (m, 4H,
Har), 7.55–7.66 (m, 1H, Har), 7.78–7.86 (m, 4H, Har),
10.08 (d, 1H, H1). 13C NMR: δ 14.88 (1C, C5), 72.56
(1C, C4), 74.23 (1C, OCH2C6H4), 126.15 (1C, C2),
127.52 (2C, Car), 128.69 (2C, Car), 130.4 (2C, Car),
Compounds 2a–c and 3a–c were tested for their abil-
ity to inhibit farnesyl transferase. None of the com-
pounds showed inhibitory activity at a concentration
up to 1 ꢂM as shown by fluorimetry testing on an iso-
lated enzyme from bovine brain [18–20].
CONCLUSION
ꢁꢁ
130.72 (2C, Car), 132.75 (1C, Car), 137.47 (1C, C1 ),
ꢁ
ꢁ
The incorporation of a second phosphorus in the
farnesyl phosphonic acid 1a did not ameliorate sig-
nificantly its activity. To make a comparison with
the benzophenone containing molecules (1b and 1c)
that were more efficiently recognized by FTase than
the farnesyl pyrophosphate, compounds 2b and 2c
have been prepared. Their biological evaluation indi-
cates that both compounds do not inhibit the FTase
at a micromolar concentration. These results may
probably be explained by a too large difference in the
polarity or/and steric properties with the substrate.
137.92 (1C, C4 ), 142.67 (1C, C1 ), 152.42 (1C, C3), 191.3
(1C, C1), 196.69 (1C, Cketone). FAB+ (NOBA): m/z 295
(M + H)+; 317 (M + Na)+. Anal calcd for C19H18O3:
C, 77.53; H, 6.16; O, 16.31. Found: C, 77.55; H, 6.14;
O, 16.30.
(E,E)-3,7-Dimethyl-8-(4ꢁ -benzoylbenzyloxy)-2,6-
octadienal (9c). (900 mg, 70%). 1H NMR: δ 1.60 (s,
3H, CH3), 1.73 (s, 3H, CH3), 2.28–2.38 (m, 4H, H4
and H5), 3.97 (s, 2H, H8), 4.56 (s, 2H, OCH2C6H4),
5.45 (m, 1H, H6), 5.92 (d, J2-1 = 7.9 Hz, 1H, H2),
7.45–7.56 (m, 4H, Har), 7.57–7.68 (m, 1H, Har), 7.78–
7.88 (m, 4H, Har), 10.02 (d, 1H, H1). 13C NMR: δ 14.41
(1C, C9), 18.10(1C, C10), 25.73 (1C, C5), 40.54 (1C,
C4), 71.45 (1C, C8), 76.71 (1C, OCH2C6H4), 126.78
(1C, C2), 127.20 (2C, Car), 127.55 (1C, C6), 128.69
(2C, Car), 130.71 (2C, Car), 130.84 (2C, Car),
EXPERIMENTAL
Reactions were monitored by TLC, using aluminum-
coated plates with silica gel 60 F254 (MERCK).
Columns chromatography were carried with Carlo
Erba silica gel 60 A (35–70 ꢂm). All solvents were
1
dried and distilled before use [21]. H and 31P NMR
132.83 (1C, C4 ), 133.82 (1C, C7), 137.16 (1C,
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
spectra were recorded with an AC 50 Bruker 200
MHz spectrometer and 13C NMR spectra with a WP-
200-SY Bruker spectrometer by using CDCl3 or D2O
as solvent. Chemical shifts are given using residual
solvent peaks as a reference relative to TMS. Mass
C1 ), 138.08 (1C, C4 ), 143.85 (1C, C1 ), 163.78 (1C,
C3), 191.62 (1C, C1), 196.96 (1C, Cketone). FAB+
(NOBA): m/z 363 (M + H)+; 385 (M + Na)+. Anal
calcd for C24H26O3: C, 79.53; H, 7.23; O, 13.24.
Found: C, 79.56; H, 7.20; O, 13.25.

658 Zgani, Menut, and Monte´ro
calcd for C24H29O5P: C, 67.28; H, 6.82; O, 18.67; P,
7.23. Found: C, 67.29; H, 6.82; O, 18.69; P, 7.24.
General Procedure for the
Horner–Wadsworth–Emmons Reaction
To a suspension of sodium hydride (2.98 mmol, 1.2
eqiv.) in dry benzene (20 ml) was added under a nitro-
gen atmosphere tetraethylmethylene diphosphonate
(2.98 mmol, 1.2 eqiv.) at room temperature. After 15
min, a solution of the aldehyde 4a, 9b, or 9c (2.48
mmol, 1 eqiv.) in dry benzene (10 ml) was added and
the mixture was stirred for 2 h. After dilution with
CH2Cl2 (100 ml), the mixture was washed with wa-
ter (100 ml), aqueous NaOH (0.1 N, 100 ml), and
water (100 ml).The organic layer was dried over an-
hydrous Na2SO4 and concentrated in vacuo. Silica
gel chromatography with ethyl acetate gave pure di-
ethylphosphonate.
(E,E,E)-[4-(9ꢁ-Diethylphosphono-2ꢁ,6ꢁ-dimethyl-2ꢁ,
6ꢁ,8ꢁ-nonatrienyloxymethyl)-phenyl]-phenyl-methanone
1
ꢁ
ꢁ
(10c). (870 mg, 75%). H NMR: δ 1.24 (t, J13 -12
=
ꢁ
ꢁ
ꢁ
ꢁ
J15 -14 = 7.0 Hz, 6H, H13 and H15 ), 1.73 (s, 3H, CH3),
ꢁ
ꢁ
1.91 (s, 3H, CH3), 2.20–2.30 (m, 4H, 2H4 and 2H5 ),
ꢁ
ꢁ
ꢁ
3.97 (s, 2H, H1 ), 4.09 (m, 4H, H12 and H14 ), 4.55
ꢁ
(s, 2H, OCH2C6H4), 5.45 (m, 1H, H3 ), 5.59 (dd,
ꢁ
ꢁ
ꢁ
ꢁ
J9 -P = 20.2 Hz, J9 -8 = 16.2 Hz, 1H, H9 ), 5.99 (d,
ꢁ
ꢁ
ꢁ
ꢁ
J7 -8 = 12.1 Hz, 1H, H7 ), 7.32–7.65 (m, 1H, H8 ),
7.46–7.65 (m, 5H, Har), 7.76–7.90 (m, 4H, Har). 31P
NMR: δ 21.7. 13C NMR: δ 14.41 (1C, C10 ), 16.78
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
(2C, C13 and C15 ), 17.75 (1C, C11 ), 26.32 (1C, C4 ),
ꢁ
ꢁ
ꢁ
ꢁ
40.11 (1C, C5 ), 62 (2C, C12 and C14 ), 71.31 (1C, C1 ),
ꢁ
76.91 (1C,OCH2C6H4), 114.94 (d, J9 -P = 192.2 Hz,
ꢁ
ꢁ
ꢁ
1C, C9 ), 124.94 (d, J7 -P = 26.56 Hz, 1C, C7 ), 127.61
(E,E,E)-1-Diethylphosphono-4,8,12-trimethyl-1,3,
7,11-tridecatetraene (5a). (526 mg, 60%; 1H NMR: δ
1.2 (t, 6H, H18,H20), 1.53 (s, 6H, H13 and H16), 1.62 (s,
3H, H15), 1.82 (s, 3H, H14), 1.93 (m, 4H, H9 and H10),
2.1 (s, 4H, H5 and H6), 4.02 (m, 4H, H17 and H19),
4.05 (m, 2H, H7 and H11), 5.48 (dd, J1-2 = 16.6 Hz,
J1-P = 20.2 Hz, 1H, H1), 5.9 (d, J3-2 = 11.2 Hz, 1H,
H3), 7.3 (ddd, J2-P = 21.0 Hz, 1H, H2); 31P NMR: δ
21.1.13C NMR: 14.78–15.10 (2C, C-18); 15.17–16.13–
16.45 (3C, C-14, C-15 and C-16); 24.47 (1C, C-13);
24.93–25.44 (1C, C-6 and C-10); 38.44–38.89 (2C, C-5
and C-9); 60.32–60.37 (2C, C-17); 112 (d, J1-P = 192.2
Hz, 1C, C-1); 121.98–122.97 (2C, C-7 and C-11);
123.07 (d, J3-P = 26.4 Hz, 1C, C-3); 130.15–134.62
(2C, C-8 and C-12); 144.21 (d, J2-P = 6.8 Hz, 1C, C-2);
147.73 (1C, C-4). FAB+ (NOBA): m/z 355 (M + H)+;
377 (M + Na)+. Anal calcd for C20H35O3P: C, 67.78;
H, 9.89; O, 13.56; P, 8.75. Found: C, 67.75; H, 9.90;
O, 13.59; P, 8.76.
ꢁ
(2C, Car), 127.83 (1C, C3 ), 128.69 (2C, Car), 130.44
ꢁꢁ
(2C, Car), 130.69 (2C, Car), 132.80 (1C, C4 ), 133.12
ꢁꢁ
(1C, C2), 137.16 (1C, C1 ), 138.11 (1C, C1), 143.84 (1C,
ꢁ
ꢁ
C4), 145.52 (d, J8 -P = 6.6 Hz, 1C, C8 ), 148.66 (1C,
C6 ), 196.89 (1C, Cketone). FAB+ (NOBA): m/z 497 (M
ꢁ
+ H)+; 519 (M + Na)+. Anal calcd for C29H37O5P: C,
70.14; H, 7.50; O, 16.11; P, 6.23. Found: C, 70.16; H,
7.49; O, 16.22; P, 6.25.
General Procedure for the Conversion of Vinyl
Phosphonates to Vinyl Phosphonic Acids
To a solution of compound 5a, 10b, or 10c (1.61
mmol, 1 eqiv.) in CH2Cl2 (30 ml) were added suc-
cessively pyridine (16.1 mmol, 10 eqiv.) and bro-
motrimethylsilane (8.1 mmol, 5 eqiv.). The mixture
was stirred for 6 h at room temperature and under ni-
trogen. The solvent was then removed and aqueous
NaOH (1 N, 30 ml) was added. The aqueous solu-
tion was stirred for 20 min then extracted with ether
to remove the pyridine excess. After acidification to
pH 2, the mixture was extracted with ether (3 × 100
ml) and the combined organic layers were washed
with brine, dried over anhydrous Na2SO4 and con-
centrated in vacuo to give pure vinyl phosphonic acid
6a, 11b, or 11c.
(E,E)-[4-(5ꢁ-Diethylphosphono-2ꢁ-methyl-2ꢁ,4ꢁ-pen-
tadienyloxymethyl)-phenyl]-phenyl-methanone (10b).
(1010 mg, 95%). 1H NMR: δ 1.34 (t, J8 -7 = J10 -9 = 6.9
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz, 6H, H8 and H10 ), 1.92 (s, 3H, CH3), 4.06 (s, 2H,
ꢁ
ꢁ
ꢁ
H1 ), 4.08–4.28 (m, 4H, 2H7 and 2H9 ), 4.60 (s, 2H,
ꢁ
ꢁ
ꢁ
OCH2C6H4), 5.70 (dd, J5 -P = 19.5 Hz, J5 -4 =16.9 Hz,
ꢁ
ꢁ
ꢁ
ꢁ
1H, H5 ), 6.26 (d, J3 -4 = 11.6 Hz, 1H, H3 ), 7.30–7.65
(m, 6H, H4 and 5Har), 7.60–7.88 (m, 4H, Har). 31P
(E,E,E)-1-Phosphono-4,8,12-trimethyl-1,3,7,11-
tridecatetraene (6a). (334 mg, 84%). 1H NMR: δ 1.61
(s, 3H, CH3), 1.70 (s, 3H, CH3), 1.83 (s, 3H, CH3),
1.94–2.17 (m, 8H), 5.02–5.21 (m, 2H), 5.58–5.76 (m,
1H), 5.91 (m, 1H), 7.12–7.5 (m, 1H), 10.6 (br s, 2H);
31P NMR: δ 21.7; 13C NMR: δ 15.66–16.43–17.70–
18.11 (4C, C13, C14, C15 and C16), 26.12–27.12 (2C,
C5 and C9), 40.10–40.54 (2C, C6 and C10), 119.53 (d,
J1-P = 184.5 Hz, 1C, C1), 123.71–124.67 (2C, C7 and
C11), 124.79 (d, J3-P = 24.6 Hz, 1C, C3), 131.79–136.23
ꢁ
13
ꢁ
NMR: δ 20.9. C NMR: δ 16.70 (1C, C6 ), 16.81 (2C,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
C8 and C10 ), 63.15 (2C, C7 et C9 ), 72.15 (1C, C1 ),
ꢁ
75.32 (1C, OCH2C6H4), 117.18 (d, J5 -P = 191.4 Hz,
ꢁ
ꢁ
ꢁ
1C, C5 ), 125.13 (d, J3 -P 26.9 Hz, 1C, C3 ), 127.54
(2C, Car), 128.72 (2C, Car), 130.50 (2C, Car), 130.74
ꢁꢁ
ꢁꢁ
(2C, Car), 132.88 (1C, C4 ), 137.37 (1C, C1 ), 138.02
ꢁ
(1C, C1), 143.20 (1C, C4), 143.99 (1C, C2 ), 144.53
(d, J4 -P = 6.7 Hz, 1C, C4 ), 196.82 (1C, Cketone). FAB+
ꢁ
(NOBA): m/z 429 (M +ꢁ H)+; 451 (M + Na)+. Anal

Synthesis of Vinyl Pyrophosphonate Analogues of Farnesyl Pyrophosphate 659
(2C, C8 and C12), 144.33 (d, J2-P = 5.9 Hz, 1C, C2),
added successively. The mixture was stirred at
room temperature under nitrogen for 2 h.
149.04 (1C, C4). FAB+ (NOBA): m/z 293 (M + H)+;
321 (M + Na)+.
• Preparation of the tri-n-butylammonium ortho-
phosphate: To a solution of orthophosphoric acid
(4.1 mmol, 3 eqiv.) in methanol (20 ml) was added
tri-n-butylamine (4.1 mmol, 3 eqiv.). The mix-
ture was stirred for 30 min at room temperature.
Methanol was evaporated under reduced pressure
and the residue was dried by repeated coevapora-
tion with dry pyridine (3 × 10 ml).
• Coupling procedure: To a stirred solution of the
tri-n-butylammonium orthophosphate (4.1 mmol,
3 eqiv.) in dry pyridine (20 ml) was added the
activated phosphonoanhydride prepared in situ
in THF within a period of 10 h at room tem-
perature and under nitrogen. The mixture was
stirred for two additional hours and then the sol-
vent was removed. Column chromatography with
isopropanol-aqueous ammonia (27%) (6:4) gave
2a–c and 3a–c.
(E,E)-[4-(5ꢁ-Phosphono-2ꢁ-methyl-2ꢁ-4ꢁ-pentadieny-
loxymethyl)-phenyl]-phenyl-methanone (11b). (460
mg, 70%). 1H NMR: δ 1.91 (s, 3H, CH3), 4.06
ꢁ
(s, 2H, H1 ), 4.60 (s, 2H, OCH2C6H4), 5.80 (dd,
ꢁ
ꢁ
ꢁ
ꢁ
J5 -P = 21.1 Hz, J5 -4 = 16.6 Hz, 1H, H5 ), 6.25 (d,
ꢁ
ꢁ
ꢁ
ꢁ
J3 -4 = 11.4 Hz, 1H, H3 ), 7.35–7.60 (m, 6H, H4 and
5Har), 7.72–7.88 (m, 4H, Har), 10.25 (s, 2H, POH).
31P NMR: δ 23.6. C NMR: δ 15.23 (1C, C6 ),
13
ꢁ
ꢁ
72.06 (1C, C1 ), 75.35 (1C, OCH2C6H4), 118.59 (d,
ꢁ
ꢁ
ꢁ
J5 -P = 188.5 Hz, 1C, C5 ), 126.65 (d, J3 -P = 26.6 Hz,
ꢁ
1C, C3 ), 127.55 (2C, Car), 128.71 (2C, Car), 130.44
ꢁꢁ
(2C, Car), 130.74 (2C, Car), 132.84 (1C, C4 ), 134.50
ꢁꢁ
ꢁ
(1C, C1 ), 135.0 (1C, C1), 137.36 (d, J4 -P = 6.1 Hz,
ꢁ
ꢁ
1C, C4 ), 143.19 (1C, C4), 144.0 (1C, C2 ), 198.0 (1C,
C
ketone). FAB+ (NOBA): m/z 373 (M + H)+; 395 (M +
Na)+.
(E,E,E)-[4-(9ꢁ-Phosphono-2ꢁ,6ꢁ-dimethyl-2ꢁ,6ꢁ,8ꢁ-
(E,E,E)-1-Pyrophosphono-4,8,12-trimethyl-1,3,7,
11-tridecatetraene (2a). (286 mg, 49%). 1H NMR:
δ 1.49 (s, 6H, 2CH3), 1.56 (s, 3H, CH3), 1.73 (s,
3H, CH3), 1.73–2.10 (m, 8H, H5, H6, H9 and H10),
4.93–5.12 (m, 2H, H7 and H9), 5.5 (m, 1H, H1), 5.98
(d, J3-2 = 11 Hz, 1H, H3), 6.88–7.13 (m, 1H, H2). 31P
NMR: δ −9.92 (d, Jꢁ–ꢀ = 22.2 Hz, 1P, Pꢁ), 8.42 (d,
1P, Pꢀ). 13C NMR: δ 15.87–16.75–16.89–17.48 (4C,
C13, C14, C15 and C16), 25.52–26.58 (2C, C5 and C9),
39.59–40.03 (2C, C6 and C10), 120.91 (d, J1-P = 188.9
Hz, 1C, C1), 124.36–124.80 (2C, C7 and C11), 125.04
(d, J3-P = 21.2 Hz, 1C, C3), 132.56–136.38 (2C, C8
and C12), 140.52 (1C, C2), 146.90 (1C, C4).
nonatrienyloxymethyl)-phenyl]-phenyl-methanone
(11c). (600 mg, 85%). 1H NMR: δ 1.72 (s, 3H,
ꢁ
CH3), 1.86 (s, 3H, CH3), 2.12–2.30 (m, 4H, 2H4 and
ꢁ
ꢁ
2H5 ), 3.96 (s, 2H, H1 ), 4.54 (s, 2H, OCH2C6H4), 5.45
ꢁ
ꢁ
ꢁ
ꢁ
(m, 1H, H3 ), 5.66 (dd, J9 -P = 20.6 Hz, J9 -8 = 16.8 Hz,
ꢁ
ꢁ
ꢁ
ꢁ
1H, H9 ), 5.95 (d, J7 -8 = 10.9 Hz, 1H, H7 ), 7.35–7.60
ꢁ
(m, 1H, H8 ), 7.40–7.68 (m, 5H, Har), 7.75–7.88 (m,
4H, Har). 31P NMR: δ 24.4. 13C NMR: δ 14.39 (1C,
ꢁ
ꢁ
ꢁ
ꢁ
C
10 ), 16.80 (1C, C11 ), 26.21 (1C, C4 ), 40.10 (1C, C5 ),
ꢁ
71.20 (1C, C9 ), 76.88 (1C, OCH2C6H4), 114.89 (d,
ꢁ
ꢁ
ꢁ
J9 -P = 190 Hz, 1C, C9 ), 124.93 (d, J7 -P = 26.3 Hz,
ꢁ
ꢁ
1C, C7 ), 127.58 (2C, Car), 127.80 (1C, C3 ), 128.67
(2C, Car), 130.47 (2C, Car), 130.71 (2C, Car), 132.81
ꢁꢁ
ꢁ
ꢁꢁ
(1C, C4 ), 133.06 (1C, C2 ), 137.04 (1C, C1 ), 138.0
(E,E,E)-1-Triphosphono-4,8,12-trimethyl-1,3,7,11-
tridecatetraene (3a). (128 mg, 18%) 1H NMR: δ 1.47
(s, 6H, 2CH3), 1.54 (s, 3H, CH3), 1.71 (s, 3H, CH3),
1.73–2.08 (m, 8H, H5, H6, H9 and H10), 4.91–5.10
(m, 2H, H7 and H9), 5.48–5.76 (m, 1H, H1), 5.97 (d,
J3-2 = 11.0 Hz, 1H, H3), 6.86–7.12 (m, 1H, H2). 31P
NMR: δ −21.3 (dd, Jꢁ–ꢀ = 23.1 Hz, Jꢁ–ꢃ = 20 Hz, Pꢁ),
−6.3 (d, Pꢃ ), 8.6 (d, Pꢀ). 13C NMR: δ 15.83–16.72–
16.91–17.52 (4C, C13, C14, C15 and C16), 25.53–26.62
(2C, C5 and C9), 39.59–40.09 (2C, C6 and C10), 120.99
(d, J1-P = 189.2 Hz, 1C, C1), 124.34–124.81 (2C,
C7 and C11), 125.14 (d, J3-P = 22.33 Hz, 1C, C3),
132.40–136.43 (2C, C8 and C12), 140.58 (1C, C2),
146.92 (1C, C4).
ꢁ
ꢁ
(1C, C1 ), 143.80 (1C, C4), 145.46 (d, J8 -P = 6.6 Hz,
1C, C8 ), 148.69 (1C, C6 ), 196.77 (1C, Cketone). FAB+
ꢁ
ꢁ
(NOBA): m/z 441 (M + H)+; 463 (M + Na)+.
General Method for the Phosphorylation
of Vinyl Phosphonic Acids
• Preparation of the activated phosphonoanhydride:
To a solution of compound 6a, 11b, or 11c (1.36
mmol, 1 eqiv.) in methanol (20 ml) was added tri-
n-butylamine (1.36 mmol, 1 eqiv.). The mixture
was stirred at room temperature for 30 min, then
methanol was evaporated under reduced pres-
sure and the residue was dried by repeated co-
evaporation with dry pyridine (3 × 10 ml). The
tri-n-butylammonium salt of the vinyl phospho-
nic acid was dissolved in dry THF (20 ml), then
diphenylphosphorochloridate (1.36 mmol, 1 eqiv.)
and tri-n-butylamine (4.1 mmol, 3 eqiv.) were
(E,E)-[4-(5ꢁ-Pyrophosphono-2ꢁ-methyl-2ꢁ,4ꢁ-penta-
dienyloxymethyl)-phenyl]-phenyl-methanone (2b).
1
(294 mg, 48%). H NMR: δ 1.64 (s, 3H, CH3), 4.0
ꢁ
(s, 2H, H1 ), 4.51 (s, 2H, OCH2C6H4), 5.90 (dd,

660 Zgani, Menut, and Monte´ro
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
3.69 (s, 2H, H1 ), 4.44 (s, 2H, OCH2C6H4), 5.36 (m,
J5 -P = 19.2 Hz, J5 -4 = 16.8 Hz, 1H, H5 ), 6.13 (d,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
J3 -4 = 10.9 Hz, 1H, H3 ), 7.10 (ddd, J4 -P = 20.4
1H, H3 ), 5.74 (dd, J9 -P = 19.6 Hz, J9 -8 = 16.9 Hz,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz, 1H, H4 ), 7.35–7.50 (m, 4H, Har), 7.54–7.62
1H, H9 ), 5.92 (d, J7 -8 = 11.3 Hz, 1H, H7 ), 7.06
(m, 1H, Har), 7.62–7.74 (m, 4H, Har). 31P NMR:
(ddd, J8 -P = 20.7 Hz, 1H, H8 ), 7.30–75.1 (m, 4H,
Har), 7.51–7.71 (m, 5H, Har). 31P NMR: δ −21.04
(dd, Jꢁ–ꢀ = 23.1 Hz, Jꢁ–ꢃ = 19.4 Hz, 1P, Pꢁ), −7.45
(d, 1P, Pꢃ ), 9.01 (d, 1P, Pꢀ). 13C NMR: δ 13.62 (1C,
ꢁ
ꢁ
δ −7.16 (d, Jꢁ–ꢀ = 23.0 Hz, 1P, Pꢁ), 6.54 (d, 1P, Pꢀ).
13
ꢁ
ꢁ
C NMR: δ 14.33 (1C, C6 ), 71.22 (1C, C1 ), 75.62
ꢁ
(1C, OCH2C6H4), 124.53 (d, J5 -P = 185.0 Hz, 1C,
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
C5 ), 127.48 (d, J3 -P = 26.2 Hz, 1C, C3 ), 128.19
C
10 ), 16.76 (1C, C11 ), 25.91 (1C, C4 ), 39.24 (1C, C5 ),
ꢁ
(2C, Car), 128.84 (2C, Car), 130.74 (2C, Car), 130.88
70.41 (1C, C1 ), 76.46 (1C, OCH2C6H4), 120.53 (d,
ꢁꢁ
ꢁꢁ
ꢁ
ꢁ
ꢁ
(2C, Car), 133.82 (1C, C4 ), 136.40 (1C, C1 ), 136.88
J9 -P = 192.5 Hz, 1C, C9 ), 124.74 (d, J7 -P = 26.3 Hz,
ꢁ
ꢁ
ꢁ
(1C, C1), 138.83 (d, J4 -P = 5.7 Hz, 1C, C4 ), 140.92 (1C,
1C, C7 ), 128.08 (2C, Car), 128.82 (2C, Car), 130.34
(2C, Car), 130.74 (2C, Car), 129.61 (1C, C3 ), 132.14
ꢁ
ꢁ
C4), 143.40 (1C, C2 ), 200.37 (1C, Cketone).
ꢁꢁ
ꢁ
ꢁꢁ
(1C, C4 ), 133.66 (1C, C2 ), 136.32 (1C, C1 ), 136.99
ꢁ
ꢁ
ꢁ
(1C, C5 ), 140.88 (d, J8 -P = 5.8 Hz, 1C, C8 ), 143.79
(E,E)-[4-(5ꢁ-Triphosphono-2ꢁ-methyl-2ꢁ,4ꢁ-penta-
ꢁ
(1C, C4), 147.03 (1C, C6 ), 199.56 (1C, Cketone).
dienyloxymethyl)-phenyl]-phenyl-methanone
(3b).
(43 mg, 6%). 1H NMR: δ 1.79 (s, 3H, CH3), 4.03 (s, 2H,
ꢁ
ꢁ
H1 ), 4.54 (s, 2H, OCH2C6H4), 5.92 (dd, J5 -P = 19.9
ꢁ
ꢁ
ꢁ
ꢁ
ꢁ
Hz, J5 -4 = 16.6 Hz, 1H, H5 ), 6.14 (d, J3 -4 = 11.1
REFERENCES
ꢁ
ꢁ
ꢁ
Hz, 1H, H3 ), 7.12 (ddd, J4 -P = 20.0 Hz, 1H, H4 ),
7.30–7.50 (m, 4H, Har), 7.52–7.68 (m, 5H, Har). 31P
NMR: δ −22.0 (dd, Jꢁ–ꢀ = 23.1 Hz, Jꢁ–ꢃ = 19.9 Hz, 1P,
Pꢁ), −7.6 (d, 1P, Pꢃ ), 9.5 (d, 1P, Pꢀ). 13C NMR: δ 14.34
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[2] Barbacid, M. Annu Rev Biochem 1987, 56, 779–827.
[3] (a) Leonhard, D. M. J Med Chem 1997, 40, 2971–2990;
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ꢁ
ꢁ
(1C, C6 ), 71.28 (1C, C1 ), 75.65 (1C, OCH2C6H4),
ꢁ
ꢁ
123.58 (d, J5 -P = 186.0 Hz, 1C, C5 ), 127.44
ꢁ
ꢁ
(d, J3 -P = 26.8 Hz, 1C, C3 ), 128.41 (2C, Car), 128.87
(2C, Car), 130.62 (2C, Car), 131.01 (2C, Car), 133.84
ꢁꢁ
ꢁꢁ
(1C, C4 ), 136.69 (1C, C1 ), 137.14 (1C, C1), 139.69
ꢁ
ꢁ
(d, J4 -P = 5.8 Hz, 1C, C4 ), 140.82 (1C, C4), 143.43
[5] Lowy, D. R.; Willumsen, B. M. Annu Rev Biochem
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ꢁ
(1C, C2 ), 201.04 (1C, Cketone).
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K.; Hohl, R. J. Bioorgan Med Chem 1998, 6, 687–694
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(E,E,E)-[4-(9ꢁ-Pyrophosphono-2ꢁ,6ꢁ-dimethyl-2ꢁ,6ꢁ,
8ꢁ-nonatrienyloxymethyl)-phenyl]-phenyl-methanone
(2c). (410 mg, 57%). H NMR: δ 1.51 (s, 3H, CH3),
[7] (a) Overhand, M.; Stuivenberg, H. R.; Pieterman, E.;
Cohen, L. H.; van Leeuwen, R. E. W.; Valentijn, A. R. P.
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(c) Cohen, L. H.; Valentijn, A. R. P. M.; Roodenburg,
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Pharmacol 1995, 49, 839–845; (d) Overhand, M.;
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I.; Distefano, M. D. Tetrahedron Lett 1996, 37, 4845–
4848.
1
ꢁ
ꢁ
1.72 (s, 3H, CH3), 1.95–2.15 (m, 4H, H4 and H5 ),
ꢁ
3.80 (s, 2H, H1 ), 4.40 (s, 2H, OCH2C6H4), 5.23–5.35
ꢁ
ꢁ
ꢁ
ꢁ
(m, 1H, H3 ), 5.78 (dd, J9 -P = 19.7 Hz, J9 -8 = 16.8
ꢁ
ꢁ
ꢁ
ꢁ
Hz, 1H, H9 ), 5.90 (d, J7 -8 = 11.1 Hz, 1H, H7 ), 7.04
ꢁ
ꢁ
(ddd, J8 -P = 20.5 Hz, 1H, H8 ), 7.28–7.46 (m, 4H,
Har), 7.51–7.63 (m, 5H, Har). 31P NMR: δ −6.45
(d, Jꢁ–ꢀ = 23.2 Hz, 1P, Pꢁ), 7.14 (d, 1P, Pꢀ). 13C
ꢁ
ꢁ
NMR: δ 13.67 (1C, C10 ), 16.75 (1C, C11 ), 25.97
ꢁ
ꢁ
ꢁ
(1C, C4 ), 39.27 (1C, C5 ), 70.46 (1C, C1 ), 76.47 (1C,
ꢁ
ꢁ
OCH2C6H4), 121.43 (d, J9 -P = 187.1 Hz, 1C, C9 ),
ꢁ
ꢁ
124.91 (d, J7 -P = 26.1 Hz, 1C, C7 ), 128.0 (2C, Car),
128.84 (2C, Car), 130.34 (2C, Car), 130.73 (2C, Car),
ꢁ
ꢁ
ꢁ
129.53 (1C, C3 ), 132.20 (1C, C4 ), 133.65 (1C, C2 ),
ꢁꢁ
136.35 (1C, C1 ), 137.04 (1C, C1), 140.13 (1C, C4),
ꢁ
ꢁ
ꢁ
143.85 (d, J8 -P = 5.6 Hz, 1C, C8 ), 146.41 (1C, C6 ),
[9] Park, H.; Bodulori, S. R.; Moomaw, J. F.; Casey, P. J.;
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(E,E,E)-[4-(9ꢁ-Triphosphono-2ꢁ,6ꢁ-dimethyl-2ꢁ,6ꢁ,
8ꢁ-nonatrienyloxymethyl)-phenyl]-phenyl-methanone
(3c). (150 mg, 17%). H NMR: δ 1.54 (s, 3H, CH3),
1
ꢁ
ꢁ
1.73 (s, 3H, CH3), 2.0–2.20 (m, 4H, H4 and H5 ),

Synthesis of Vinyl Pyrophosphonate Analogues of Farnesyl Pyrophosphate 661
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