U.S. patent application number 10/690914 was filed with the patent office on 2005-04-28 for bicyclic carbohydrate compounds useful in the treatment of infections caused by herpesviridae.
Invention is credited to Blom, Petra, Eycken, Johan Van der, hemel, Johan Van, Peys, Eric, Ruttens, Bart, Sas, Benedikt, Vandenkerckhove, Jan.
Application Number | 20050090452 10/690914 |
Document ID | / |
Family ID | 34521749 |
Filed Date | 2005-04-28 |
United States Patent
Application |
20050090452 |
Kind Code |
A1 |
Sas, Benedikt ; et
al. |
April 28, 2005 |
Bicyclic carbohydrate compounds useful in the treatment of
infections caused by herpesviridae
Abstract
Bicyclic carbohydrates for the treatment of infections caused by
herpseviridae, and in particular cytomegalovirus. The invention
consists of the novel bicyclic carbohydrates the generic structure
of which is: 1 wherein R.sub.1 is either -Bn or -Ph; R.sub.2 and
R.sub.3 are either -alkyl, -aryl, -allyl, or --H; R.sub.4 and
R.sub.5 form a ring and are either --CH(Ph)- or --CH(aryl)- and X
is either O, N or S.
Inventors: |
Sas, Benedikt; (Stekene,
BE) ; hemel, Johan Van; (Antwerpen, BE) ;
Vandenkerckhove, Jan; (Zichem, BE) ; Peys, Eric;
(Balen, BE) ; Eycken, Johan Van der; (Ninove,
BE) ; Ruttens, Bart; (Gent, BE) ; Blom,
Petra; (Sint-Amandsberg, BE) |
Correspondence
Address: |
DAVIS, BROWN, KOEHN, SHORS & ROBERTS, P.C.
THE FINANCIAL CENTER
666 WALNUT STREET
SUITE 2500
DES MOINES
IA
50309-3993
US
|
Family ID: |
34521749 |
Appl. No.: |
10/690914 |
Filed: |
October 22, 2003 |
Current U.S.
Class: |
514/25 ; 514/327;
514/432; 536/120; 546/242; 549/28 |
Current CPC
Class: |
C07H 7/02 20130101; C07D
309/10 20130101; C07H 15/203 20130101; C07D 493/04 20130101; A61P
31/04 20180101; C07H 7/04 20130101; A61P 31/10 20180101; A61P 33/02
20180101; C07H 9/04 20130101; A61P 31/12 20180101; C07H 15/04
20130101; C07H 15/10 20130101; A61P 35/00 20180101; A61P 43/00
20180101 |
Class at
Publication: |
514/025 ;
514/327; 514/432; 536/120; 546/242; 549/028 |
International
Class: |
A61K 031/70; C07H
015/04; A61K 031/382; A61K 031/445; C07D 211/36 |
Claims
We claim:
1. A compound of the formula: 12wherein: R.sub.1 is selected from
the group consisting of -alkyl and -aryl; R.sub.2 and R.sub.3 are
selected from the group consisting of -alkyl, -aryl, -allyl and
--H; R.sub.4 and R.sub.5 form a ring and are selected from the
group consisting of --CH(Ph)- and --CH(aryl)-; X is selected from
the group consisting of O, N and S; or a pharmaceutically active
derivative thereof.
2. A method as defined in claim 1, wherein R.sub.1 preferably is
selected from the group consisting of phenyl and benzyl; R.sub.2
and R.sub.3 are preferably selected from the group consisting of
-methyl, -ethyl, -allyl, -propargyl and hydrogen; R.sub.4 and
R.sub.5 form a ring and are selected preferably from the group
consisting of --CH(Ph)-, --CH(naphtyl)- and --CH(biphenyl)-; and X
is preferably O; or a pharmaceutically active derivative
thereof.
3. A method of treating a pathogenic viral infection in a mammalian
subject comprising the step of administering to the subject a
composition comprising at least one compound of claim 1.
4. The method of claim 3 wherein the composition contains a
compound of claim 1 in an effective anti-viral amount.
5. The method of claim 3 wherein the mammalian subject is a human
patient or another mammal.
6. A method for treating a pathogenic viral infection in a
mammalian subject where the infective agent is resistant to one or
more other therapies, comprising the step of administering to the
subject a composition comprising an effective anti-viral amount of
a compound of claim 1.
7. A method as defined in claim 3, wherein the viral infection is
an infection caused by herpesviridae.
8. A method as defined in claim 8, wherein the viral infection is
an infection caused by cytomegalovirus.
Description
BACKGROUND OF THE INVENTION
[0001] The invention relates generally to compounds active against
viral diseases and, more specifically to bicyclic carbohydrate
compounds that are useful in the prophylaxis and treatment of
diseases caused by the alphaherpesvirina cytomegalovirus.
[0002] Cytomegalovirus (CMV) is the largest human herpes virus, and
there is only one serotype. As with animal CMVs it is species
specific; humans are the natural hosts and animal CMVs do not
infect humans. The name refers to the multinucleated cells, which
together with the intranuclear inclusions, are characteristic
responses to infection with this virus. Transmission is via saliva,
and CMVs were originally called `salivary gland` viruses. Urine is
an additional source of infection in children, and in infected
pregnant women the virus can spread via the blood to the placenta
and fetus. Semen and cervical secretions may also contain virus and
it can therefore be spread by sexual contact. It is often present
in milk in small quantities, but this is of doubtful significance
in transmission. In hospitals it can be transmitted by blood
transfusion and organ transplants. CMV infections are often
asymptomatic, but can reactivate and cause disease when CMI (cell
mediated immune response) defenses are impaired. After clinical
silent infection of unknown cells in the upper respiratory tract,
CMV spreads locally to lymphoid tissues and then systemically in
circulating lymphocytes and monocytes to involve lymph nodes and
the spleen. The infection then localizes in epithelial cells in
salivary glands and kidney tubules, and in cervix, testes and
epididymis, from where the virus is shed to the outside world
(Table 1).
1TABLE 1 Cytomegalovirus infections Site of infection Result
Comment Salivary glands Salivary transmission Importance of kissing
and contaminated hands Tubular Virus in urine Probable role of
transmission epithelium of Sexual transmission Up to 10.sup.7
infectious doses/ml kidney Cervix/ of semen in an acutely Testis/
infected male epididymis Lymphocytes/ Virus spread through Probable
site of persistent Macrophages body via infected cells infection
Mononucleosis may occur Immunosuppressive effect Placenta/Fetus
Congenital abnormalities Greatest damage in fetus after primary
maternal infection rather than reactivation
[0003] Infected cells may be multinucleated or bear intranuclear
inclusions, but pathologic changes are minor and infection is
generally asymptomatic. In young adults, a glandular fever type
illness can occur, but without heterophil antibodies. There is
fever and lethargy, and abnormal lymphocytes and mononucleosis in
blood smears. The virus inhibits T cell responses and there is a
temporary reduction in their immune reactivity to other antigens.
Although specific antibodies and CMI responses are generated, these
fail to clear the virus, which often continues to be shed in saliva
and urine for many months. The infection is, however, eventually
controlled by CMI mechanisms, although infected cells remain in the
body throughout life and can be a source of reactivation and
disease when CMI defenses are impaired. CMV owes its success in our
species to its ability to evade immune defenses (Cann A. J. (1997)
Principles of Molecular Virology. Second Edition. Academic Press,
San Diego, pp. 65-67; Mims C., Playfair J., Roitt I., Wakelin D.
and Williams R. (1998) Medicinal Microbiology. Second Edition.
Mosby International Limited, pp.347-348).
[0004] Ganciclovir is an anti-CMV drug that works by inhibiting
CMV's DNA polymerase enzyme. It can prevent CMV from reproducing
and infecting new cells, but it cannot eliminate it from the body.
Ganciclovir is manufactured by Roche under the trade name Cymevene.
In the US its trade name is Cytovene. Ganciclovir now comes in
several formulations: oral, intravenous and intravitreal (into the
eye). It is either given intravenously on a long-term basis through
a catheter or in a pill form. In the case of administration by
catheter, one end of a tube is surgically inserted into a large
vein in the chest, the other end of the tube remains outside the
chest or has an injectable port just under the skin. Although
slightly less efficient, an oral form of ganciclovir is also
approved by the FDA for prevention and maintenance treatment of
CMV.
[0005] Unfortunately, ganciclovir also suppresses bone-marrow
production of the white blood cells called neutrophils. This
condition is called neutropenia. People taking ganciclovir require
close monitoring to ensure blood disorders are promptly detected.
It can also harm the kidneys, reduce testosterone levels, and cause
nausea, vomiting, diarrhea and rash. More than 10% of IV
ganciclovir recipients have to stop treatment because of these side
effects. In addition, the development of strains of CMV that are
resistant to ganciclovir has been reported. These do not seem to
develop any faster with oral ganciclovir than with the intravenous
formulation (www.aidsmap.com).
[0006] Alternative compounds with activity against herpes viruses
such as cytomegalovirus are needed.
SUMMARY OF THE INVENTION
[0007] The invention consists of the novel bicyclic carbohydrates
the generic structure of which is: 2
[0008] wherein R.sub.1 is either -Bn or -Ph; R.sub.2 and R.sub.3
are either -alkyl, -aryl, -allyl, or --H; R.sub.4 and R.sub.5 form
a ring and are either --CH(Ph)- or --CH(aryl)- and X is either O, N
or S, herein referred to as Formula A. Compounds of Formula A have
activity against infections caused by the cytomegalovirus. The
invention also includes analogs, prodrugs and pharmaceutically
acceptable salts thereof, together with pharmaceutical compositions
for the prophylaxis and treatment of diseases caused by infections
of alphaherpesvirinae and particularly cytomegalovirus.
BRIEF DESCRIPTION OF THE DRAWINGS
[0009] FIG. 1 is a chemical structure of the bicyclic carbohydrates
of the present invention and designated Formula A.
[0010] FIG. 2 is a diagrammatic representation of the activity of
ganciclovir in cells that are non-infected and cells that are
infected with herpes virus.
[0011] FIG. 3 is a diagrammatic representation of the scheme of
synthesis of Compound A1.
[0012] FIG. 4 is a diagrammatic representation of the scheme of
synthesis of Compound A2.
[0013] FIG. 5 is a diagrammatic representation of the scheme of
synthesis of Compound A3.
[0014] FIG. 6 is a diagrammatic representation of the scheme of
synthesis of Compound A4.
[0015] FIG. 7 is a diagrammatic representation of the scheme of
synthesis of Compound A5.
[0016] FIG. 8 is a diagrammatic representation of the scheme of
synthesis of Compounds A6 and A7.
DETAILED DESCRIPTION OF A PREFERRED EMBODIMENT
[0017] The inventive compounds may be used in their native form or
as salts. In cases where compounds are sufficiently basic or acidic
to form stable nontoxic acid or base salts, administration of the
compounds as salts may be appropriate. Examples of pharmaceutically
acceptable salts are organic acid addition salts formed with acids
which form a physiological acceptable anion, for example, acetate
ascorbate, benzoate, citrate, etoglutarate, glycerophosphate,
malonate, methanesulfonate, succinate, and tartarate. Suitable
inorganic salts may also be formed, including bicarbonate,
carbonate, hydrochloride, nitrate, and sulfate, salts.
[0018] Compounds of the present invention can conveniently be
administered in a pharmaceutical composition containing the
compound in combination with a suitable excipient, the composition
being useful in combating viral infections. Depending on whether
the preparation is used to treat internal or external viral
infections, the compounds and compositions of the present invention
can be administered parenterally, topically, intravaginally,
orally, or rectally.
[0019] For parenteral administration, solutions of the active
compound or its salts can be prepared in water, optionally mixed
with a nontoxic surfactant. Dispersions can also be prepared in
glycerol, liquid polyethylene glycols, triacetin, and mixtures
thereof and in oils.
[0020] Useful dosages of the compound can be determined by
comparing their in vitro activity. Methods for the extrapolation of
effective dosages to humans are known to the art.
[0021] The compound is conveniently administered in unit dosage
form; for example, containing 0.1 to 2000 mg, conveniently 100 to
1000 mg, most conveniently, 100 to 500 mg of active ingredient per
unit dosage form. The desired dose may conveniently be presented in
a single dose or as divided doses administered at appropriate
intervals, for example, as two, three, four or more sub-doses per
day. The sub-dose itself may be further divided, e.g., into a
number of discrete loosely spaced administrations; such as multiple
inhalations from an insufflator or by application of a plurality of
drops into the eye.
[0022] For internal infections, the compositions can be
administered orally or parenterally at dose levels, calculated as
the free base, of about 1 to 30 mg/kg, preferably 1 to 30 mg/kg of
mammal body weight, and can be used in man in a unit dosage form,
administered one to four times daily in the amount of 1 to 10 mg
per unit dose.
[0023] For parenteral administration or for administration as
drops, as for eye infections, the compounds are presented in
aqueous solution in a concentration of from about 0.1 to about 20%,
more preferably about 1 to about 5%. The solution may contain other
ingredients, such as emulsifiers, antioxidants or buffers.
[0024] The exact regimen for administration of the compound and
compositions disclosed herein will necessarily be dependent upon
the needs of the individual subject being treated, the type of
treatment and, of course, the judgment of the attending
practitioner. The compounds of the present invention can be
administered to an animal in need of treatment. In most instances,
this will be a human being, but the treatment of livestock and
companion animals is also specifically contemplated as falling
within the scope of the instant invention.
[0025] Compounds of Formula A and pharmaceutically acceptable salts
thereof are useful as antiviral agents. Thus, they are useful to
combat viral infections in animals, including man. The compounds
are generally active against herpes viruses, and are particularly
useful in the prophylaxis and treatment of diseases caused by the
alphaherpesvirinae cytomegalovirus.
[0026] Methods and Materials
[0027] The structures of the bicyclic carbohydrates synthesized are
presented in Table 2.
2TABLE 2 Structures of active compounds Compound Structure R.sub.1
R.sub.2 R.sub.3 R.sub.4 R.sub.5 Compound A1 3 --Ph --OMe --OMe
--OCH(Ph)OCH.sub.2-- Compound A2 4 --Bn --OMe --OMe
--OCH(Ph)OCH.sub.2-- Compound A3 5 --Bn --OEt --OEt
--OCH(Ph)OCH.sub.2-- Compound A4 6 --Bn --OAll --OAll
--OCH(Ph)OCH.sub.2-- Compound A5 7 --Bn --OCH.sub.2CCH
--OCH.sub.2CCH --OCH(Ph)OCH.sub.2-- Compound A6 8 --Ph --OMe --OMe
9 Compound A7 10 --Ph --OMe --OMe 11
EXAMPLE 1
Synthesis of the Compounds of Formula A
[0028] The compounds were synthesized as follows:
[0029] 1. Synthesis of Compound A1
[0030] The scheme of the synthesis of Compound A1 is illustrated in
FIG. 3.
[0031] Synthesis of Compound 1.1
[0032] To (.beta.)-D-glucose penta-acetate (24.6 g, 63.0 mmol) was
added a solution of hydrogen bromide in acetic acid (33 wt %, 100
ml). A dark brown color immediately appears. The reaction mixture
was stirred at room temperature for 30 minutes under argon
atmosphere. Subsequently the solvent was removed by azeotropic
distillation in vacuo with toluene (4.times.50 ml), yielding a
green-brown solid Compound 1.1. The crude product was used in the
next reaction step without further purification.
[0033] Formula: C.sub.14H.sub.19O.sub.9Br
[0034] Molecular weight: 411.20
[0035] R.sub.f: 0.46 (cyclohexane/ethyl acetate 1:1)
[0036] IR (KBr): 2962, 2360, 2342, 1748, 1435, 1369, 1218, 1162,
1112, 1079, 1042, 911, 752, 668, 601, 563 cm.sup.-1
[0037] ES-MS: 433=[410+Na].sup.+, 435=[412+Na.sup.+]
[0038] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 6.61 (1H, d,
J=4.0 Hz), 5.56 (1H, dd, app. t, J=9.7 Hz), 5.16 (1H, dd, app. t,
J=9.7 Hz), 4.84 (1H, dd, J=10.0, 4.0 Hz), 4.33 (1H, m), 4.30 (1H,
m), 4.13 (1H, dd, J=12.3, 1.5 Hz), 2.11 (3H, s), 2.10 (3H, s), 2.05
(3H, s), 2.03 (3H, s)
[0039] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 170.37, 169.70,
169.64, 169.31, 86.34, 71.91, 70.39, 69.94, 66.94, 60.76, 20.48,
20.48, 20.38, 20.38
[0040] Synthesis of Compound 1.2
[0041] To a solution of phenylmagnesium bromide (200 ml of a 3M
solution in diethyl ether, 600 mmol, 9.5 eq) in dry diethyl ether
(500 ml), cooled to 0.degree. C., was added a solution of the
bromide Compound 1.1 (63.0 mmol theoretical) in dry diethyl ether
(500 ml) by canulation. The reaction mixture was stirred at room
temperature under argon-atmosphere for 72 hours. Subsequently the
reaction mixture was poured out into water (2000 ml), and acetic
acid (200 ml) was added to dissolve the magnesium-salts. The two
layers were separated, and the organic layer was washed with water
(3.times.500 ml). The combined aqueous layers were concentrated
under reduced pressure to yield a light brown solid residue. This
residue was dissolved in pyridine (500 ml). At 0.degree. C. acetic
anhydride (340 ml) was added slowly. After adding DMAP (200 mg,
1.64 mmol), stirring was continued for 20 hours at room temperature
under argon-atmosphere. Next the reaction mixture was concentrated
under reduced pressure, followed by azeotropic distillation with
toluene (1.times.250 ml), and the addition of diethyl ether (3 l).
The obtained organic layer was washed with sat. NaHCO.sub.3-sol.
(2.times.1 l), 1 N HCl-sol. (2.times.1 l) and water (2.times.1 l).
Drying on MgSO.sub.4, and concentrating under reduced pressure,
yielded 25.1 g light brown crystals. These were purified by
recrystallization from 2-propanol, to give 16.1 g Compound 1.2
(63%) as white crystals.
[0042] Formula: C.sub.20H.sub.24O.sub.9
[0043] Molecular weight: 408.40
[0044] R.sub.f: 0.42 (cyclohexane/ethyl acetate 1:1)
[0045] Melting point: 149-150.degree. C.
[0046] IR (KBr): 2956, 1753, 1433, 1368, 1224, 1104, 1036, 978,
916, 764, 738, 702, 603 cm.sup.-1
[0047] ES-MS: 431=[408+Na].sup.+.sup.1H-NMR (500 MHz, CDCl.sub.3):
.delta. 7.39 (5H, m), 5.24 (1H, dd, app. t, J=9.4 Hz), 5.24 (1H,
dd, app. t, J=9.8 Hz), 5.14 (1H, dd, app. t, J=9.8 Hz), 4.40 (1H,
d, J=9.9 Hz), 4.30 (1H, dd, J=17.2, 4.7 Hz), 4.16 (1H, dd, J=12.2,
1.5 Hz), 3.85 (1H, m), 2.09 (3H , s), 2.06 (3H, s), 2.01 (3H, s),
1.80 (3H, s)
[0048] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 170.60, 170.25,
169.36, 168.70, 136.01, 128.75, 128.28, 126.96, 80.08, 75.94,
74.06, 72.44, 68.39, 62.17, 20.61, 20.48, 20.21
[0049] Synthesis of Compound 1.3
[0050] To a solution of the tetra-acetate, Compound 1.2, (16.08 g,
39.4 mmol) in a mixture of tetrahydrofuran (232 ml) and methanol
(232 ml) was added anhydric potassium carbonate (1.36 g, 9.84 mmol,
0.25 eq). The mixture was stirred at room temperature under
argon-atmosphere for 3 hours. Silicagel (40 ml) was added and the
solvent was removed under reduced pressure. Purification of the
product Compound 1.3 by column chromatography
(dichloromethane/methanol 85/15) gives 9.50 g of product Compound
1.3 (99%).
[0051] Formula: C.sub.12H.sub.16O.sub.5
[0052] Molecular weight: 240.26
[0053] R.sub.f: 0.12 (dichloromethane/methanol 9:1)
[0054] IR (KBr): 3368, 2919, 2360, 1636, 1496, 1455, 1082, 1042,
891, 764, 701, 595 cm.sup.-1
[0055] ES-MS: 258=[240+NH.sub.4].sup.+, 263=[240+Na].sup.+
[0056] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.44 (2H, d,
J=7.1 Hz), 7.35 (2H, dd, app. t, J=7.6 Hz), 7.30 (1H, m), 4.15 (1H,
d, J=9.4 Hz), 3.90 (1H, dd, J=12.1, 1.6 Hz), 3.72 (1H, dd, J=12.0,
5.2 Hz), 3.51 (1H, dd, app. t, J=8.7 Hz), 3.45 (1H, dd, app. t,
J=9.4 Hz), 3.43 (3H, m), 3.40 (1H, dd, app. t, J=9.2 Hz)
[0057] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 139.30, 127.43,
82.41, 80.70, 78.23, 74.98, 70.40, 61.41
[0058] Synthesis of Compound 1.4
[0059] To a solution of tetrol, Compound 1.3 (1.15 g, 4.79 mmol) in
dry acetonitrile (3 ml) under argon-atmosphere was added
camphorsulfonic acid (279 mg, 1.20 mmol, 0.25 eq) and benzaldehyde
dimethyl acetal (1.44 ml, 9.58 mmol, 2 eq). The reaction mixture
was stirred at room temperature for 3 hours. Subsequently the
mixture was neutralized by addition of triethylamine (0.337 ml,
2.40 mmol). Concentrating the reaction mixture under reduced
pressure yields 2.70 g of a light yellow oil. Purification by
column chromatography (CH.sub.2Cl.sub.2/iPrOH 1/1) gives 1.53 g of
Compound 1.4 (97%) as a white solid.
[0060] Formula: C.sub.19H.sub.20O.sub.5
[0061] Molecular weight: 328.36
[0062] R.sub.f: 0.27 (cyclohexane/ethyl acetate 1:1)
[0063] Melting point: 114-115.degree. C.
[0064] [.alpha.].sub.D.sup.20=+9.3.degree.;
[.alpha.].sub.365.sup.20=+10.0- .degree. (c=1.13 in chloroform)
[0065] IR (KBr): 3433, 2874, 2357, 1651, 1496, 1455, 1385, 1313,
1272, 1211, 1109, 1029, 1009, 913, 765, 733, 700 cm.sup.-1
[0066] ES-MS: 346=[328+NH.sub.4].sup.+
[0067] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.53 (2H, m),
7.40 (5H, m), 7.39 (3H, m), 5.59 (1H, s), 4.37 (1H, dd, J=10.3, 5.9
Hz), 4.30 (1H, d, J=9.3 Hz), 3.91 (1H, dd, app. t, J=8.6 Hz) 3.79
(1H, dd, app. t, J=10.3 Hz), 3.67 (1H, dd, app. t, J=9.3 Hz), 3.65
(1H, m), 3.63 (1H, m)
[0068] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 137.50, 136.84,
129.14, 128.65, 128.52, 128.20, 127.29, 126.12, 101.73, 82.41,
80.90, 75.43, 74.60, 70.60, 68.70
[0069] C,H-analysis: theoretical: C, 69.50%; H, 6.14%; found: C,
70.79%; H, 6.11%.
[0070] Synthesis of Compound A1
[0071] To a solution of the diol, Compound 1.4 (2.5 g, 7.6 mmol) in
dry dimethyl ethylene glycol (80 ml), cooled to 0.degree. C., was
added sodium hydride (913 mg, 22.850 mmol). The reaction mixture
was stirred at 0.degree. C. under argon atmosphere for 30 minutes.
Subsequently methyl iodide (1.6 ml, 25.12 mmol) was added. The
reaction mixture was stirred at room temperature under
argon-atmosphere for 2 hours. The reaction mixture was poured out
into water (170 ml), followed by separation of layers, and
extraction of the aqueous layer with diethyl ether (3.times.350
ml). The combined organic layers were dried over magnesium sulfate
and the solvent was removed under reduced pressure to yield 3.0 g
(99%) of Compound A1 as a yellow solid.
[0072] Formula: C.sub.21H.sub.24O.sub.5
[0073] Molecular weight: 356.42
[0074] R.sub.f: 0.59 (cyclohexane/ethyl acetate 8:2)
[0075] Melting point: 97-98.degree. C.
[0076] [.alpha.].sub.D.sup.20=-21.0.degree.;
[.alpha.].sub.365.sup.20=67.0- .degree. (c=0.80 in chloroform)
[0077] IR (KBr): 3035, 2925, 2851, 2360, 1497, 1455, 1378, 1276,
1173, 1104, 1030, 999, 959, 916, 765, 699 cm.sup.-1
[0078] ES-MS: 357=[356+H].sup.+
[0079] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.53 (2H, m),
7.40 (3H, m), 7.38 (5H, m), 5.60 (1H, s), 4.37 (1H, dd, J=10.4, 4.9
Hz), 4.26 (1H, d, J=9.5 Hz), 3.78 (1H, dd, app. t, J=10.2 Hz), 3.69
(1H, dd, J=9.4, 9.2 Hz), 3.68 (3H, s), 3.58 (1H, ddd, J=9.9, 9.9,
4.9 Hz), 3.55 (1H, dd, J=9.1, 9.0 Hz), 3.20 (1H, dd, app. t, J=9.5,
8.5 Hz), 3.07 (3H, s)
[0080] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 138.51, 137.19,
128.77, 128.23, 128.06, 127.25, 125.88, 100.99, 85.28, 84.25,
82.17, 81.86, 70.16, 68.78, 60.84, 60.58
[0081] C,H-analysis: theoretical: C, 70.77%; H, 6.79%; found: C,
73.68%; H, 8.40%.
[0082] 2. Synthesis of Compound A2
[0083] The scheme of the synthesis of Compound A2 is illustrated in
FIG. 4
[0084] Synthesis of Compound 2.1
[0085] To a solution of 2,3,4,6-tetra-O-benzyl-D-glucopyranose 7.35
(4.0 g, 7.4 mmol) in dry methylene chloride (50 ml) and
dimethylformamide (2.5 ml) was added at room temperature a solution
of oxalyl bromide (1 ml, 10 mmol) in dry methylene chloride (1 ml).
The mixture was stirred at room temperature for 1 hour and then
poured into ice water (50 ml). The two layers were separated and
the organic layer was washed with cold water (2.times.50 ml), dried
(MgSO.sub.4) and filtered. The solvent was removed under reduced
pressure to an orange-colored oil. The crude product Compound 2.1
was used in the next step.
[0086] Formula: C.sub.34H.sub.35BrO.sub.5
[0087] Molecular weight: 603.55
[0088] R.sub.f: 0.53 (cyclohexane/ethyl acetate 85:15)
[0089] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.37 (3H, m),
7.33 (5H, m), 7.31 (5H, m), 7.28 (5H, m), 7.15 (2H, m), 6.43 (1H,
d, J=3.7 Hz), 4.98 (1H, d, J=5.0 Hz), 4.83 (2H, dd, app. t, J=10.9
Hz), 4.58 (1H, d, J=12.1 Hz), 4.50 (1H, d, J=10.7 Hz), 4.46 (2H, d,
J=4.06 (1H, m), 4.03 (1H, dd, app. t, J=9.2 Hz), 3.80 (1H, m), 3.78
(1H, m), 3.76 (1H, d, J=4.6 Hz), 3.65 (1H, dd, J=11.0, 2.0 Hz),
3.54 (1H, dd, J=9.2, 3.7 Hz)
[0090] Synthesis of compound 2.2
[0091] To a solution of the bromide, Compound 2.1 (4.47 g, 7.4
mmol) in dry diethyl ether (100 ml) was added at 0.degree. C.
benzyl magnesium bromide (60 ml of a 1M solution in diethyl ether,
60 mmol). The mixture was stirred at 0.degree. C. for 1 hour and at
room temperature for 18 hours. Then the reaction mixture was poured
into water (200 ml) and acetic acid (10 ml) was added. The two
layers were separated and the organic layer was washed with a
saturated sodium bicarbonate solution (3.times.250 ml) and brine
(2.times.250 ml). The organic layer was dried (MgSO.sub.4),
filtered and the solvent was removed under reduced pressure. The
crude product was purified by flash chromatography (gradient
elution:cyclohexane/ethyl acetate 95/5 to 85/15), followed by HPLC
(eluent:cyclohexane/diethyl ether 9/1) to yield 2.2 g (48%) of a
colorless oil Compound 2.2.
[0092] Formula: C.sub.41H.sub.42O.sub.5
[0093] Molecular weight: 614.78
[0094] R.sub.f: 0.15 (cyclohexane/diethyl ether 9:1)
[0095] [.alpha.].sub.D.sup.20=+85.3; [.alpha.].sub.365.sup.20=+88.1
(c=0.60 in chloroform)
[0096] IR (KBr): 2862, 2360, 1604, 1496, 1454, 1360, 1209, 1085,
1028, 735, 697, 668 cm.sup.-1
[0097] ES-MS: 632=[614+NH.sub.4].sup.+
[0098] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.36 (5H, m),
7.34 (5H, m), 7.31 (5H, m), 7.29 (5H, m), 7.26 (2H, m), 7.22 (3H,
m), 4.96 (1H, d, J=11.0 Hz), 4.95 (1H, d, J=11.0 Hz), 4.91 (1H, d,
J=11.0 Hz), 4.84 (1H, d, J=10.8 Hz), 4.69 (1H, d, J=11.0 Hz), 4.62
(1H, d, J=10.8 Hz), 4.59 (1H, d, J=12.2 Hz), 4.52 (1H, d, J=12.2
Hz), 3.74 (1H, dd, app. t, J=9.0 Hz), 3.69 (1H, m), 3.68 (1H, m),
3.66 (1H, dd, app. t, J=9.3 Hz), 3.52 (1H, ddd, J=18.3, 9.2, 2.3
Hz), 3.37 (1H, dd, app. t, J=9.0 Hz), 3.36 (1H, m), 3.17 (1H, dd,
J=14.3, 2.0 Hz), 2.75 (1H, dd, J=14.3, 8.8 Hz)
[0099] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 138.68, 138.41,
138.22, 138.04, 138.01, 129.49, 128.35, 128.31, 128.26, 128.15,
127.91, 127.77, 127.69, 127.56, 127.50, 127.33, 125.95, 87.26,
81.59, 79.86, 78.80, 78.47, 75.41, 74.99, 74.81, 73.22, 68.77,
37.72
[0100] C,H-analysis: calculated: C, 80.10%; H, 6.90%; found: C,
79.38%; H, 7.09%.
[0101] Synthesis of compound 2.3
[0102] To a solution of Compound 2.2 (2.0 g, 3.25 mmol) in ethanol
(80 ml) was added at room temperature palladium on carbon (Pd--C,
200 mg). The mixture was shaken (Parr apparatus) at room
temperature for 2 hours under a hydrogen pressure of 4 atm. The
suspension was filtered over celite, the filter was washed with
ethanol and tetrahydrofuran, and the solvent was removed under
reduced pressure. The crude product was purified by flash
chromatography (eluent:methylene chloride/methanol 9/1) to yield
1.15 g (99%) of product Compound 2.3.
[0103] Formula: C.sub.13H.sub.18O.sub.5
[0104] Molecular weight: 254.28
[0105] R.sub.f: 0.14 (dichloromethane/methanol 9:1)
[0106] IR (KBr): 3381, 2922, 2360, 2341, 1641, 1603, 1496, 1454,
1379, 1308, 1226, 1079, 1031, 897, 754, 701, 668 cm.sup.-1
[0107] ES-MS: 272=[254+NH.sub.4].sup.+
[0108] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.29 (2H, d,
J=7.0 Hz), 7.22 (2H, dd, app. t, J=7.3 Hz), 7.14 (1H, m), 3.75 (1H,
dd, J=11.9, 2.4 Hz), 3.60 (1H, dd, J=11.8, 5.4 Hz), 3.35 (1H, m),
3.32 (1H, m), 3.25 (1H, dd, app. t, J=9.4 Hz), 3.15 (1H, m), 3.12
(1H, m), 3.09 (1H, dd, app. t, J=9.3 Hz), 2.69 (1H, dd, J=14.5, 8.5
Hz)
[0109] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 140.50, 130.71,
128.96, 126.97, 81.73, 81.40, 79.91, 74.91, 71.90, 62.98, 38.73
[0110] C,H-analysis: calculated: C, 61.40%; H, 7.10%; found: C,
58.92%; H, 7.15%.
[0111] Synthesis of Compound 2.4
[0112] To a solution of tetrol, Compound 2.3, (1.0 g, 3.93 mmol) in
dimethylformamide (40 ml) were added at room temperature
camphorsulfonic acid (274 mg, 1.18 mmol) en benzaldehyde dimethyl
acetal (0.652 ml, 4.72 mmol). The mixture was stirred at
110.degree. C. for 6 hours. It was then diluted with ethyl acetate
(100 ml), washed with a 1M sodium hydroxide solution (2.times.150
ml), a saturated sodium bicarbonate solution (2.times.150 ml) and
brine (2.times.150 ml). The organic layer was dried (MgSO.sub.4),
filtered and the solvent was removed under reduced pressure. The
crude product was purified by flash chromatography (gradient
elution:cyclohexane/ethyl acetate 9/1 to 6/4) to yield 950 mg (71%)
of a white solid, Compound 2.4.
[0113] Formula: C.sub.20H.sub.22O.sub.5
[0114] Molecular weight: 342.39
[0115] R.sub.f: 0.20 (cyclohexane/ethyl acetate 6:4)
[0116] Melting point: 43-44.degree. C.
[0117] [.alpha.].sub.D.sup.20=-6.9; [.alpha.].sub.365.sup.20=-10.7
(c=0.60 in chloroform)
[0118] IR (KBr): 3478, 3031, 2871, 2360, 1604, 1497, 1454, 1385,
1317, 1299, 1271, 1212, 1124, 1099, 1077, 998, 973, 919, 673, 699,
668, 655, 625, 552, 510 cm.sup.-1
[0119] ES-MS: 343=[342+H].sup.+
[0120] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.49 (2H, m),
7.38 (3H, m), 7.31 (2H, m), 7.28 (2H, m), 7.25 (1H, m), 5.51 (1H,
s), 4.28 (1H, dd, J=10.5, 4.8 Hz), 3.74 (1H, dd, app. t, J=8.7 Hz),
3.68 (1H, dd, app. t, J=10.0 Hz), 3.58 (1H, ddd, J=9.6, 8.2, 2.6
Hz), 3.43 (1H, dd, app. t, J=9.2 Hz), 3.39 (1H, m), 3.38 (1H, dd,
J=10.5, 4.0 Hz), 3.18 (1H, dd, J=14.4, 2.5 Hz), 2.93 (1H, bs), 2.79
(1H, dd, J=14.4, 7.9 Hz), 2.69 (1H, bs)
[0121] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 137.84, 136.93,
129.63, 129.20, 128.26, 128.02, 126.20, 126.15, 101.69, 80.90,
80.14, 75.24, 73.58, 69.94, 68.71, 37.71
[0122] C,H-analysis: calculated: C, 70.20%; H, 6.50%; found: C,
68.84%; H, 6.59%.
[0123] Synthesis of Compound A2
[0124] To a solution of diol Compound 2.4 (920 mg, 2.69 mmol) in
dry dimethyl ethylene glycol (30 ml) was added at 0.degree. C.
sodium hydride (650 mg, 16.12 mmol). The mixture was stirred at
0.degree. C. for 30 minutes. Iodomethane (0.67 ml, 10.75 mmol) was
added at 0.degree. C. and the reaction mixture was stirred at room
temperature for 16 hours. It was then poured into water (50 ml) and
the two layers were separated. The water layer was extracted with
ethyl acetate (3.times.50 ml). The combined organic layers were
dried (MgSO.sub.4), filtered and the solvent was removed under
reduced pressure. The crude product was purified by flash
chromatography (gradient elution:cyclohexane/ethyl acetate 95/5 to
7/3) to yield 907 mg (91%) of a white solid, Compound A2.
[0125] Formula: C.sub.22H.sub.26O.sub.5
[0126] Molecular weight: 370.44
[0127] R.sub.f: 0.63 (cyclohexane/ethyl acetate 6:4)
[0128] Melting point: 103-104.degree. C.
[0129] IR (KBr): 3027, 2982, 2891, 2831, 1603, 1496, 1455, 1380,
1323, 1277, 1232, 1167, 1141, 1121, 1094, 1030, 989, 958, 875, 754,
698, 654, 622, 580, 543, 502 cm.sup.-1
[0130] ES-MS: 371=[370+H].sup.+
[0131] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.49 (2H, m),
7.35 (3H, m), 7.29 (2H, m), 7.25 (3H, m), 5.53 (1H, s), 4.25 (1H,
dd, J=10.5, 5.0 Hz), 3.67 (1H, dd, J=10.3, 5.0 Hz), 3.66 (3H, s),
3.63 (3H, s), 3.51 (1H, m), 3.49 (1H, dd, app. t, J=8.8 Hz), 3.47
(1H, m), 3.30 (1H, ddd, J=14.4, 9.5, 5.0 Hz), 3.14 (1H, dd, J=14.4,
2.1 Hz), 2.98 (1H, dd, app. t, J=8.8 Hz), 2.73 (1H, dd, J=14.3, 8.4
Hz)
[0132] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 138.20, 137.32,
129.51, 128.74, 128.06, 127.98, 126.10, 125.89, 100.92, 84.92,
82.83, 82.10, 80.23, 69.88, 68.77, 60.77, 60.59, 37.97
[0133] C,H-analysis: calculated: C, 71.30%; H, 7.10%; found: C,
71.26%; H 7.45%.
[0134] 3. Synthesis of Compound A3
[0135] The scheme of the synthesis of Compound A3 is illustrated in
FIG. 5.
[0136] To a solution of diol, Compound 2.4 of the scheme of FIG. 4,
(150 mg, 0.438 mmol) in dry DMF (2.2 ml) was added at 0.degree. C.
sodium_hydride (42 mg, 1.752 mmol). The mixture was stirred at
0.degree. C. for 15 minutes. Ethyl bromide (75 .mu.l, 1.007 mmol)
was added at 0.degree. C. and the reaction mixture was stirred at
room temperature overnight. The reaction was quenched by adding
MeOH. The mixture was concentrated under reduced pressure. Ether
(30 ml) and water (30 ml) were added, followed by separation of
layers. Extraction of the aqueous layers with ether (3.times.25
ml), washing the combined organic layers with brine (50 ml), drying
on MgSO.sub.4 and concentrating under reduced pressure yielded 180
mg solid residue. This was purified by column chromatography
(230-400 mesh silica, cyclohexane:ethyl acetate 96:4), to give 130
mg of Compound A3. (75%)
[0137] Formula: C.sub.24H.sub.30O.sub.5
[0138] Molecular weight: 398.49
[0139] R.sub.f: 0.20 (cyclohexane/ethyl acetate 95:5)
[0140] Melting point: 87-89.degree. C.
[0141] IR (KBr): 2973 (m), 2868 (m), 1496 (m), 1454 (m), 1374 (m),
1276 (m), 1170 (m), 1142 (m), 1087 (s), 1029 (m), 1008 (m), 698
(m)
[0142] ES-MS: 399=[M+H].sup.+
[0143] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.48 (2H, m),
7.36 (3H, m), 7.25 (5H, m), 5.52 (1H, s), 4.24 (1H, dd, J=10.4, 5.0
Hz), 3.98 (2H, m), 3.75 (1H, m), 3.67 (2H, m), 3.54 (2H, m), 3.49
(1H, dd, app. t, J=9.1 Hz), 3.29 (1H, ddd, app. dt, J=9.9, 5.0 Hz),
3.15 (1H, dd, J=14.3, 2.1 Hz), 3.09 (1H, dd, app. t, J=9.1 Hz),
2.72 (1H, dd, J=14.3, 8.6 Hz), 1.24 (3H, t, J=7.1 Hz), 1.22 (3H, t,
J=7.0 Hz)
[0144] APT-NMR (125 MHz, CDCl.sub.3): .delta. 138.6 (C), 137.6 (C),
129.7 (CH), 128.9 (CH), 128.3 (CH), 128.2 (CH), 126.3 (CH), 126.1
(CH), 101.1 (CH), 83.4 (CH), 82.2 (CH), 81.8 (CH), 80.7 (CH), 70.3
(CH), 69.0 (CH.sub.2), 68.9 (CH.sub.2), 68.6 (CH.sub.2), 38.3
(CH.sub.2), 16.0 (CH.sub.3), 15.9 (CH.sub.3)
[0145] 4. Synthesis of Compound A4
[0146] The scheme of the synthesis of Compound A4 is illustrated in
FIG. 6.
[0147] To a solution of diol, Compound 2.4 of the scheme of FIG. 4,
(250 mg, 0.730 mmol) in dry DMF (3.5 ml) was added at 0.degree. C.
sodium hydride (70 mg, 2.92 mmol). The mixture was stirred at
0.degree. C. for 20 minutes. Allyl bromide (145 .mu.l, 1.679 mmol)
was added at 0.degree. C. and the reaction mixture was stirred at
room temperature overnight. The reaction was quenched by adding
MeOH. Ether (30 ml) and water (30 ml) were added, followed by
separation of layers. Extraction of the aqueous layers with ether
(3.times.30 ml), washing the combined organic layers with brine (50
ml), drying on MgSO.sub.4 and concentrating under reduced pressure
yielded 314 mg solid residue. This was purified by column
chromatography (230-400 mesh silica, cyclohexane:ethyl acetate
96:4), to give 231 mg of Compound A4. (76%)
[0148] Formula: C.sub.26H.sub.30O.sub.5
[0149] Molecular weight: 422.51
[0150] R.sub.f: 0.69 (cyclohexane/ethyl acetate 1:1)
[0151] Melting point: 58-60.degree. C.
[0152] IR (KBr): cm.sup.-1 2867 (m), 1454 (m), 1383 (m), 1171 (m),
1101 (s), 1084 (s), 1033 (m), 1000 (m), 972 (m), 921 (m), 747 (m),
691 (s)
[0153] ES-MS: 423=[M+H].sup.+
[0154] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. (ppm) 7.47 (2H,
dd, J=7.7, 1.7 Hz), 7.38-7.34 (3H, m), 7.30-7.20 (5H, m), 6.00-5.91
(2H, m), 5.52 (1H, s), 5.32 (1H, dd, J=17.2, 1.6 Hz), 5.28 (1H, dd,
J=17.2, 1.7 Hz), 5.20 (1H, dd, J=10.4, 1.6 Hz), 5.16 (1H, dd,
J=10.4, 1.7 Hz), 4.47 (1H, dd, J=12.3, 5.6 Hz), 4.43 (1H, dd,
J=12.6, 5.6 Hz), 4.26-4.21 (2H, m), 4.15 (1H, dd, J=12.3, 5.8 Hz),
3.66 (1H, dd, app. t, J=10.3 Hz), 3.65 (1H, dd, app. t, J=9.0 Hz),
3.57 (1H, ddd, app. dt, J=9.4, 2.3 Hz), 3.54 (1H, dd, app. t, J=9.3
Hz), 3.31 (1H, ddd, app. dt, J=9.9, 5.0 Hz), 3.17 (2H, m), 2.72
(1H, dd, J=14.4, 8.5 Hz)
[0155] APT-NMR (125 MHz, CDCl.sub.3): .delta. (ppm) 138.4 (C),
137.5 (C), 135.2 (CH), 134.9 (CH), 129.7 (CH), 128.9 (CH), 128.3
(CH), 128.2 (CH), 126.3 (CH), 126.1 (CH), 117.1 (CH.sub.2), 116.9
(CH.sub.2), 101.1 (CH), 83.1 (CH), 82.3 (CH), 81.2 (CH), 80.5 (CH),
74.4 (CH.sub.2), 73.9 (CH.sub.2), 70.2 (CH), 69.0 (CH.sub.2), 38.2
(CH.sub.2)
[0156] 5. Synthesis of Compound A5
[0157] The scheme of the synthesis of Compound A5 is illustrated in
FIG. 7.
[0158] Synthesis of Compound 5.1
[0159] To a solution of 2,3,4,6-tetra-O-benzyl-D-glucopyranose (4.0
g, 7.4 mmol) in dry methylene chloride (50 ml) and
dimethylformamide (2.5 ml) was added at room temperature a solution
of oxalyl bromide (1 ml, 10 mmol) in dry methylene chloride (1 ml).
The mixture was stirred at room temperature for 1 hour and then
poured into ice-water (50 ml). The two layers were separated and
the organic layer was washed with cold water (2.times.50 ml), dried
(MgSO.sub.4) and filtered. The solvent was removed under reduced
pressure to an orange colored oil. The crude product Compound 5.1
was used in the next step.
[0160] Formula: C.sub.34H.sub.35BrO.sub.5
[0161] Molecular weight: 603.55
[0162] R.sub.f: 0.53 (cyclohexane/ethyl acetate 85:15)
[0163] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.37 (3H, m),
7.33 (5H, m), 7.31 (5H, m), 7.28 (5H, m), 7.15 (2H, m), 6.43 (1H,
d, J=3.7 Hz), 4.98 (1H, d, J=5.0 Hz), 4.83 (2H, dd, app. t, J=10.9
Hz), 4.58 (1H, d, J=12.1 Hz), 4.50 (1H, d, J=10.7 Hz), 4.46 (2H, d,
J=12.1 Hz), 4.06 (1H, m), 4.03 (1H, dd, app. t, J=9.2 Hz), 3.80
(1H, m), 3.78 (1H, m), 3.76 (1H, d, J=4.6 Hz), 3.65 (1H, dd,
J=11.0, 2.0 Hz), 3.54 (1H, dd, J=9.2, 3.7 Hz)
[0164] Synthesis of Compound 5.2
[0165] To a solution of bromide 5.1 (4.47 g, 7.4 mmol) in dry
diethyl ether (100 ml) was added at 0.degree. C. benzylmagnesium
bromide (60 ml of a 1M solution in diethyl ether, 60 mmol). The
mixture was stirred at 0.degree. C. for 1 hour and at room
temperature for 18 hours. Then the reaction mixture was poured into
water (200 ml) and acetic acid (10 ml) was added. The two layers
were separated and the organic layer was washed with a saturated
sodium bicarbonate solution (3.times.250 ml) and brine (2.times.250
ml). The organic layer was dried (MgSO.sub.4), filtered and the
solvent was removed under reduced pressure. The crude product was
purified by flash chromatography (gradient
elution:cyclohexane/ethyl acetate 95/5 to 85/15), followed by HPLC
(eluens:cyclohexane/diethyl ether 9/1) to yield 2.2 g (48%) of a
colorless oil Compound 5.2.
[0166] Formula: C.sub.41H.sub.42O.sub.5
[0167] Molecular weight: 614.78
[0168] R.sub.f: 0.15 (cyclohexane/diethyl ether 9:1)
[0169] [.alpha.].sub.D.sup.20=+85.3; [.alpha.].sub.365
.sup.20=+88.1 (c=0.60 in chloroform)
[0170] IR (KBr): 2862, 2360, 1604, 1496, 1454, 1360, 1209, 1085,
1028, 735, 697, 668 cm.sup.-1
[0171] ES-MS: 632=[614+NH.sub.4].sup.+
[0172] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.36 (5H, m),
7.34 (5H, m), 7.31 (5H, m), 7.29 (5H, m), 7.26 (2H, m), 7.22 (3H,
m), 4.96 (1H, d, J=11.0 Hz), 4.95 (1H, d, J=11.0 Hz), 4.91 (1H, d,
J=11.0 Hz), 4.84 (1H, d, J=10.8 Hz), 4.69 (1H, d, J=11.0 Hz), 4.62
(1H, d, J=10.8 Hz), 4.59 (1H, d, J=12.2 Hz), 4.52 (1 H, d, J=12.2
Hz), 3.74 (1H, dd, app. t, J=9.0 Hz), 3.69 (1H, m), 3.68 (1H, m),
3.66 (1H, dd, app. t, J=9.3 Hz), 3.52 (1H, ddd, J=18.3, 9.2, 2.3
Hz), 3.37 (1H, dd, app. t, J=9.0 Hz), 3.36 (1H, m), 3.17 (1H, dd,
J=14.3, 2.0 Hz), 2.75 (1H, dd, J=14.3, 8.8 Hz)
[0173] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 138.68, 138.41,
138.22, 138.04, 138.01, 129.49, 128.35, 128.31, 128.26, 128.15,
127.91, 127.77, 127.69, 127.56, 127.50, 127.33, 125.95, 87.26,
81.59, 79.86, 78.80, 78.47, 75.41, 74.99, 74.81, 73.22, 68.77,
37.72
[0174] C,H-analysis: calculated: C, 80.10%; H, 6.90%; found: C,
79.38%; H, 7.09%.
[0175] Synthesis of Compound 5.3
[0176] To a solution of Compound 5.2 (2.0 g, 3.25 mmol) in ethanol
(80 ml) was added at room temperature palladium on carbon (Pd--C,
200 mg). The mixture was shaken (Parr apparatus) at room
temperature for 2 hours under a hydrogen pressure of 4 atm. The
suspension was filtered over celite, the filter was washed with
ethanol and tetrahydrofuran, and the solvent was removed under
reduced pressure. The crude product was purified by flash
chromatography (eluent:methylene chloride/methanol 9/1) to yield
1.15 g (99%) of product Compound 5.3.
[0177] Formula: C.sub.13H.sub.18O.sub.5
[0178] Molecular weight: 254.28
[0179] R.sub.f: 0.14 (dichloromethane/methanol 9:1)
[0180] IR (KBr): 3381, 2922, 2360, 2341, 1641, 1603, 1496, 1454,
1379, 1308, 1226, 1079, 1031, 897, 754, 701, 668 cm.sup.-1
[0181] ES-MS: 272=[254+NH.sup.4].sup.+
[0182] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.29 (2H, d,
J=7.0 Hz), 7.22 (2H, dd, app. t, J=7.3 Hz), 7.14 (1H, m), 3.75 (1H,
dd, J=11.9, 2.4 Hz), 3.60 (1H, dd, J=11.8, 5.4 Hz), 3.35 (1H, m),
3.32 (1H, m), 3.25 (1H, dd, app. t, J=9.4 Hz), 3.15 (1H, m), 3.12
(1H, m), 3.09 (1H, dd, app. t, J=9.3 Hz), 2.69 (1H, dd, J=14.5, 8.5
Hz)
[0183] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 140.50, 130.71,
128.96, 126.97, 81.73, 81.40, 79.91, 74.91, 71.90, 62.98, 38.73
[0184] C,H-analysis: calculated: C, 61.40%; H, 7.10%; found: C,
58.92%; H, 7.15%.
[0185] Synthesis of Comnpound 5.4
[0186] To a solution of tetrol Compound 5.3 (1.0 g, 3.93 mmol) in
dimethylformamide (40 ml) were added at room temperature
camphorsulfonic acid (274 mg, 1.18 mmol) en benzaldehyde dimethyl
acetal (0.652 ml, 4.72 mmol). The mixture was stirred at
110.degree. C. for 6 hours. It was then diluted with ethyl acetate
(100 ml), washed with a 1M sodium hydroxide solution (2.times.150
ml), a saturated sodium bicarbonate solution (2.times.150 ml) and
brine (2.times.150 ml). The organic layer was dried (MgSO.sub.4),
filtered and the solvent was removed under reduced pressure. The
crude product was purified by flash chromatography (gradient
elution:cyclohexane/ethyl acetate 9/1 to 6/4) to yield 950 mg (71%)
of a white solid Compound 5.4.
[0187] Formula: C.sub.20H.sub.22O.sub.5
[0188] Molecular weight: 342.39
[0189] R.sub.f: 0.20 (cyclohexane/ethyl acetate 6:4)
[0190] Melting point: 43-44.degree. C.
[0191] [.alpha.].sub.D.sup.20=-6.9; [.alpha.].sub.365.sup.20=-10.7
(c=0.60 in chloroform)
[0192] IR (KBr): 3478, 3031, 2871, 2360, 1604, 1497, 1454, 1385,
1317, 1299, 1271, 1212, 1124, 1099, 1077, 998, 973, 919, 673, 699,
668, 655, 625, 552, 510 cm.sup.-1
[0193] ES-MS: 343=[342+H].sup.+
[0194] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.49 (2H, m),
7.38 (3H, m), 7.31 (2H, m), 7.28 (2H, m), 7.25 (1H, m), 5.51 (1H,
s), 4.28 (1H, dd, J=10.5, 4.8 Hz), 3.74 (1H, dd, app. t, J=8.7 Hz),
3.68 (1H, dd, app. t, J=10.0 Hz), 3.58 (1H, ddd, J=9.6, 8.2, 2.6
Hz), 3.43 (1H, dd, app. t, J=9.2 Hz), 3.39 (1H, m), 3.38 (1H, dd,
J=10.5, 4.0 Hz), 3.18 (1H, dd, J=14.4, 2.5 Hz), 2.93 (1H, bs), 2.79
(1H, dd, J=14.4, 7.9 Hz), 2.69 (1H, bs)
[0195] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 137.84, 136.93,
129.63, 129.20, 128.26, 128.02, 126.20, 126.15, 101.69, 80.90,
80.14, 75.24, 73.58, 69.94, 68.71, 37.71
[0196] C,H-analysis: calculated: C, 70.20%; H, 6.50%; found: C,
68.84%; H 6.59%.
[0197] Synthesis of Compound A5
[0198] To a solution of diol Compound 5.4 (80 mg, 0.234 mmol) in
dry THF (410 .mu.L) was added at 0.degree. C. sodium hydride (23 mg
of a 60% dispersion, 2.2 eq). The mixture was heated to
reflux-temperature and stirred as such for 1.5 h. After cooling the
reaction mixture to 0.degree. C., tetra-n-butylammonium iodide (1.8
mg, 0.02 eq) and propargyl bromide (57 .mu.l, 2.2 eq) were added.
The reaction mixture is stirred at room temperature overnight,
after which starting material is still present, so extra portions
of sodium hydride (13 mg 60% dispersion, 1.1 eq) and
tetra-n-butylammonium iodide (1.8 mg, 0.02 eq) were added. Stirring
was continued at room temperature during 2.5 h. Work-up was started
by pouring out in water (50 ml), followed by extraction with
diethyl ether (3.times.40 ml). The combined organic layers were
washed with brine (50 ml) and dried over MgSO.sub.4. Filtration and
concentration in vacuo yielded 104 mg crude product, which was
purified by column chromatography (230-400 mesh silica,
pentane:ether 85:15). This gave 83 mg Compound A5 as a white
crystalline product. (86%)
[0199] Formula: C.sub.26H.sub.26O.sub.5
[0200] Molecular weight: 418.48
[0201] R.sub.f: 0.26 (pentane:ether 85:15)
[0202] Melting point: 81-83.degree. C.
[0203] [.alpha.].sub.D.sup.20=-45.0.degree.;
[.alpha.].sub.365.sup.20=-132- .80.degree. (c=1.02 in
chloroform)
[0204] IR (KBr): 3288 (s), 3037 (m), 2931 (m), 2872 (s), 1454 (m),
1355 (m), 1102 (s), 1085 (s), 1032 (s), 1008 (s), 973 (m), 750 (m),
697 (s), 656 (m), 636 (m)
[0205] ES-MS: 419=[M+H].sup.+
[0206] EI-MS: (m/z) 69 (23), 91 (100), 105 (58), 149 (17), 171
(10), 221 (2), 247 (2), 269 (2), 327 (3), 379 (11) [M.sup.+-39]
[0207] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.47-7.45 (2H,
m), 7.39-7.35 (3H, m), 7.31-7.21 (5H, m), 5.51 (1H, s), 4.62 (1H,
dd, J=15.7, 2.4 Hz), 4.52 (1H, dd, J=15.7, 2.4 Hz), 4.50 (1H, dd,
J=15.7, 2.4 Hz), 4.43 (1H, dd, J=15.7, 2.4 Hz), 4.25 (1H, dd,
J=10.5, 5.0 Hz), 3.83 (1H, dd, J=9.1, 8.6 Hz), 3.67 (1H, dd, app.
t, J=10.3 Hz), 3.58 (1H, dd, app. t, J=9.4 Hz), 3.55 (1H, ddd, app.
dt, J=9.0, 2.2 Hz), 3.36-3.27 (3H, m), 2.73 (1H, dd, J=14.5, 8.7
Hz), 2.50 (1H, t, J=2.4 Hz), 2.48 (1H, t, J=2.4 Hz)
[0208] APT-NMR (125 MHz, CDCl.sub.3): .delta. 138.4 (C), 137.2 (C),
129.7 (CH), 129.0 (CH), 128.3 (CH), 128.1 (CH), 126.3 (CH), 126.0
(CH), 101.1 (CH), 82.6 (CH), 82.2 (CH), 80.1 (CH), 79.5 (CH), 74.7
(C), 74.5 (C), 69.8 (CH), 68.8 (CH.sub.2), 60.2 (CH.sub.2), 59.7
(CH.sub.2), 38.1 (CH.sub.3)
[0209] 6. Synthesis of compound A6.
[0210] The scheme of the synthesis of Compound A6 is illustrated in
FIG. 8.
[0211] Synthesis of Compound 6.1
[0212] To (.beta.)-D-glucose penta-acetate (24.6 g, 63.0 mmol) was
added a solution of hydrogen bromide in acetic acid (33 wt %, 100
ml). A dark-brown color immediately appears. The reaction mixture
was stirred at room temperature for 30 minutes under argon
atmosphere. Subsequently the solvent was removed by azeotropic
distillation in vacuo with toluene (4.times.50 ml), yielding a
green-brown solid Compound 6.1. The crude product was used in the
next reaction step without further purification.
[0213] Formula: C.sub.14H.sub.19O.sub.9Br
[0214] Molecular weight: 411.20
[0215] R.sub.f: 0.46 (cyclohexane/ethyl acetate 1:1)
[0216] IR (KBr): 2962, 2360, 2342, 1748, 1435, 1369, 1218, 1162,
1112, 1079, 1042, 911, 752, 668, 601, 563 cm.sup.-1
[0217] ES-MS: 433=[410+Na].sup.+, 435=[412+Na.sup.+]
[0218] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 6.61 (1H, d,
J=4.0 Hz), 5.56 (1H, dd, app. t, J=9.7 Hz), 5.16 (1H, dd, app. t,
J=9.7 Hz), 4.84 (1H, dd, J=10.0, 4.0 Hz), 4.33 (1H, m), 4.30 (1H,
m), 4.13 (1H, dd, J=12.3, 1.5 Hz), 2.11 (3H, s), 2.10 (3H, s), 2.05
(3H, s), 2.03 (3H, s)
[0219] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 170.37, 169.70,
169.64, 169.31, 86.34, 71.91, 70.39, 69.94, 66.94, 60.76, 20.48,
20.48, 20.38, 20.38
[0220] Synthesis of Compound 6.2
[0221] To a solution of phenylmagnesium bromide (200 ml of a 3M
solution in diethyl ether, 600 mmol, 9.5 eq) in dry diethyl ether
(500 ml), cooled to 0.degree. C., was added a solution of bromide
Compound 6.1 (63.0 mmol theoretical) in dry diethyl ether (500 ml)
by canulation. The reaction mixture was stirred at room temperature
under argon-atmosphere for 72 hours. Subsequently the reaction
mixture was poured out into water (2000 ml), and acetic acid (200
ml) was added to dissolve the magnesium-salts. The two layers were
separated, and the organic layer was washed with water (3.times.500
ml). The combined aqueous layers were concentrated under reduced
pressure to yield a light-brown solid residue. This residue was
dissolved in pyridine (500 ml). At 0.degree. C. acetic anhydride
(340 ml) was added slowly. After adding DMAP (200 mg, 1.64 mmol),
stirring was continued for 20 hours at room temperature under
argon-atmosphere. Next the reaction mixture was concentrated under
reduced pressure, followed by azeotropic distillation with toluene
(1.times.250 ml), and the addition of diethyl ether (3 l). The
obtained organic layer was washed with sat. NaHCO.sub.3-sol.
(2.times.1 l), 1 N HCl-sol. (2.times.1 l) and water (2.times.1 l).
Drying on MgSO.sub.4, and concentrating under reduced pressure,
yielded 25.1 g light-brown crystals. These were purified by
recrystallization from 2-propanol, to give 16.1 g Compound 6.2
(63%) as white crystals.
[0222] Formula: C.sub.20H.sub.24O.sub.9
[0223] Molecular weight: 408.40
[0224] R.sub.f: 0.42 (cyclohexane/ethyl acetate 1:1)
[0225] Melting point: 149-150.degree. C.
[0226] IR (KBr): 2956, 1753, 1433, 1368, 1224, 1104, 1036, 978,
916, 764, 738, 702, 603 cm.sup.-1
[0227] ES-MS: 431=[408+Na].sup.+
[0228] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.39 (5H, m),
5.24 (1H, dd, app. t, J=9.4 Hz), 5.24 (1H, dd, app. t, J=9.8 Hz),
5.14 (1H, dd, app. t, J=9.8 Hz), 4.40 (1H, d, J=9.9 Hz), 4.30 (1H,
dd, J=17.2, 4.7 Hz), 4.16 (1H, dd, J=12.2, 1.5 Hz), 3.85 (1H, m),
2.09 (3H, s), 2.06 (3H, s), 2.01 (3H, s), 1.80 (3H, s)
[0229] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 170.60, 170.25,
169.36, 168.70, 136.01, 128.75, 128.28, 126.96, 80.08, 75.94,
74.06, 72.44, 68.39, 62.17, 20.61, 20.48, 20.21
[0230] Synthesis of Compound 6.3
[0231] To a solution of tetra-acetate Compound 6.2 (16.08 g, 39.4
mmol) in a mixture of tetrahydrofuran (232 ml) and methanol (232
ml) was added anhydric potassium carbonate (1.36 g, 9.84 mmol, 0.25
eq). The mixture was stirred at room temperature under
argon-atmosphere for 3 hours. Silicagel (40 ml) was added and the
solvent was removed under reduced pressure. Purification of the
product Compound 6.3 by column chromatography
(dichloromethane/methanol 85/15) gives 9.50 g of product 6.3
(99%).
[0232] Formula: C.sub.12H.sub.16O.sub.5
[0233] Molecular weight: 240.26
[0234] R.sub.f: 0.12 (dichloromethane/methanol 9:1)
[0235] IR (KBr): 3368, 2919, 2360, 1636, 1496, 1455, 1082, 1042,
891, 764, 701, 595 cm.sup.-1
[0236] ES-MS: 258=[240+NH.sub.4].sup.+, 263=[240+Na].sup.+
[0237] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.44 (2H, d,
J=7.1 Hz), 7.35 (2H, dd, app. t, J=7.6 Hz), 7.30 (1H, m), 4.15 (1H,
d, J=9.4 Hz), 3.90 (1H, dd, J=12.1, 1.6 Hz), 3.72 (1H, dd, J=12.0,
5.2 Hz), 3.51 (1H, dd, app. t, J=8.7 Hz), 3.45 (1H, dd, app. t,
J=9.4 Hz), 3.43 (3H, m), 3.40 (1H, dd, app. t, J=9.2 Hz)
[0238] .sup.13C-NMR (125 MHz, CDCl.sub.3): .delta. 139.30, 127.43,
82.41, 80.70, 78.23, 74.98, 70.40, 61.41
[0239] Synthesis of Compound 6.4
[0240] To a solution of tetrol Compound 6.3 (300 mg, 1.25 mmol) in
dry acetonitrile (12 ml) under argon-atmosphere were added
camphorsulfonic acid (30 mg, 0.1 eq), anhydr. cupper(II) sulfate
(300 mg, 1.5 eq), and 2-naphtaldehyde (975 mg, 5 eq). The reaction
mixture was heated to reflux temperature. After stirring as such
for 12 h the reaction was poured out into water (50 ml), followed
by extraction of the aqueous layer with dichloromethane (3.times.50
ml). The combined organic layers were washed with brine (50 ml),
neutralized with triethylamine and dried on MgSO.sub.4. Filtration
and concentration in vacuo gave 370 mg residue which was purified
by column chromatography (230-400 mesh silica,
dichloromethane:isopropanol 98:2) and HPLC
(dichloromethane:isopropanol 100:2.5, UV-detection at 280 nm). This
yielded 318 mg pure Compound 6.4 as a solid (67%).
[0241] Formula: C.sub.23H.sub.22O.sub.5
[0242] Molecular weight: 378.42
[0243] R.sub.f: 0.28 (dichloromethane/isopropanol 97.5/2.5)
[0244] Melting point: 211-212.quadrature. C
[0245] [.alpha.].sub.D.sup.20=-18.6.degree.;
[.alpha.].sub.365.sup.20=-60.- 3.degree. (c=1.00 in methanol)
[0246] IR (KBr): 3412 (m), 2872 (m), 1455 (m), 1390 (m), 1373 (m),
1344 (m), 1267 (m), 1173 (m), 1126 (s), 1107 (s), 1073 (s), 1032
(m), 1009 (s), 973 (m), 897 (m), 861 (m), 820 (m), 761 (m), 744
(m), 701 (s), 579 (m), 479 (m) cm.sup.-1
[0247] EI-MS: (m/z) 91 (38), 128 (38), 156 (100), 378 (35)
[M.sup.+]
[0248] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 8.03 (1H, s),
7.92 (3H, m), 7.65 (1H, dd, J=8.5, 1.6 Hz), 7.52 (2H, m), 7.41 (2H,
br d, J=7.0 Hz), 7.30 (3H, m), 5.80 (1H, s), 4.64 (1H, br s), 4.34
(1H, d, J=9.5 Hz), 4.28 (1H, dd, J=10.2, 4.8 Hz), 4.24 (1H, br s),
3.82 (1H, dd, app. t, J=10.2 Hz), 3.81 (1H, m), 3.68 (1H, dd, app.
t, J=9.1 Hz), 3.63 (1H, ddd, J=10.0, 9.3, 4.8 Hz), 3.54 (1H, br t,
J=8.9 Hz)
[0249] APT-NMR (125 MHz, CDCl.sub.3): .delta. 140.7 (C), 136.7 (C),
134.5 (C), 133.8 (C), 129.1 (CH), 128.7 (CH), 128.6 (CH), 128.5
(CH), 128.5 (CH), 127.2 (CH), 127.0 (CH), 126.5 (CH), 125.2 (CH),
102.2 (CH), 83.6 (CH), 82.6 (CH), 76.9 (CH), 76.0 (CH), 71.7 (CH),
69.5 (CH.sub.2)
[0250] Synthesis of Compound A6
[0251] To a solution of Compound 6.4 (150 mg, 0.396 mmol) in dry
DMF (3.96 ml), cooled to 0.degree. C., was added sodium hydride (65
mg of a 60% dispersion, 4 eq). The reaction mixture was stirred at
0.degree. C. under argon atmosphere for 30 minutes. Subsequently
methyl iodide (125 .mu.l, 5 eq) was added. The reaction mixture was
stirred at room temperature under argon-atmosphere for 15 hours.
The reaction mixture was poured out into water (50 ml), followed by
separation of layers, and extraction of the aqueous layer with
diethyl ether (3.times.50 ml). The combined organic layers were
washed with brine (50 ml), dried over magnesium sulfate and the
solvent was removed under reduced pressure. The residue was
purified by column chromatography (230-400 mesh silica,
pentane:dichloromethane:et- her 10:10:0.25), to yield 149 mg
Compound A6 as a white solid. (93%).
[0252] Formula: C.sub.25H.sub.26O.sub.5
[0253] Molecular weight: 406.47
[0254] R.sub.f: 0.24 (pentane:dichloromethane:ether 10:10:0.25)
[0255] Melting point: 135.degree. C.
[0256] [.alpha.].sub.D.sup.20=-27.6.degree.;
[.alpha.].sub.365.sup.20=-103- .7.degree. (c=1.03 in
chloroform)
[0257] IR (KBr): 2932 (m), 2896 (m), 2834 (m), 1174 (m), 1142 (m),
1104 (s), 1088 (s), 1044 (m), 1033 (m), 1003 (m), 959 (m), 858 (m),
822 (m), 764 (m), 700 (m) cm.sup.-1
[0258] EI-MS: (m/z) 45 (19), 88 (45), 121 (100), 156 (53), 199
(22), 406 (9) [M.sup.+]
[0259] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 8.00 (1H, s),
7.88-7.84 (3H, m), 7.63 (1H, dd, J=8.5, 1.6 Hz), 7.50-7.48 (2H, m),
7.42-7.32 (5H, m), 5.76 (1H, s), 4.42 (1H, dd, J=10.4, 5.0 Hz),
4.28 (1H, d, J=9.6 Hz), 3.84 (1H, dd, app. t, J=10.2 Hz), 3.75 (1H,
dd, app. t, J=9.3 Hz), 3.70 (3H, s), 3.62 (1H, ddd, app. dt, J=9.8,
5.0 Hz), 3.58 (1H, dd, app. t, J=9.3 Hz), 3.22 (1H, dd, J=9.5, 8.6
Hz), 3.08 (3H, s)
[0260] APT-NMR (125 MHz, CDCl.sub.3): .delta. 138.8 (C), 134.9 (C),
133.7 (C), 133.0 (C), 128.5 (CH), 128.1 (CH), 127.7 (CH), 127.5
(CH), 126.4 (CH), 126.2 (CH), 125.6 (CH), 123.8 (CH), 101.5 (CH),
85.6 (CH), 84.6 (CH), 82.5 (CH), 82.3 (CH), 70.8 (CH), 69.2
(CH.sub.2), 61.0 (CH.sub.3), 60.8 (CH.sub.3)
[0261] 7. Synthesis of Compound A7
[0262] The scheme of the synthesis of Compound A7 is illustrated in
FIG. 8.
[0263] Synthesis of Compound 7.1
[0264] To a solution of tetrol Compound 6.3 (300 mg, 1.25 mmol) in
dry acetonitrile (12 ml) under argon-atmosphere were added
camphorsulfonic acid (30 mg, 0.1 eq), anhydr. cupper(II) sulfate
(300 mg, 1.5 eq), and 4-biphenylcarboxaldehyde (1.15 g, 5 eq). The
reaction mixture was heated to reflux temperature. After stirring
as such for 24 h the reaction was poured out into water (50 ml),
followed by extraction of the aqueous layer with dichloromethane
(3.times.50 ml). The combined organic layers were washed with brine
(50 ml), neutralized with triethylamine and dried on MgSO.sub.4.
Filtration and concentration in vacuo gave 370 mg residue which was
purified by column chromatography (230-400 mesh silica, gradient:
pure dichloromethane to dichloromethane:isopropanol 98:2). This
yielded 360 mg pure Compound 7.1 as a solid (71%).
[0265] Formula: C.sub.25H.sub.24O.sub.5
[0266] Molecular weight: 404.46
[0267] R.sub.f: 0.22 (dichloromethane/isopropanol 98/2)
[0268] Melting point: 195-197.degree. C.
[0269] [.alpha.].sub.D.sup.20=-0.49.degree.;
[.alpha.].sub.365.sup.20=-8.0- 2.degree. (c=1.03 in chloroform)
[0270] IR (KBr): 3412 (m), 2867 (m), 1385 (m), 1108 (s), 1077 (s),
1033 (m), 1008 (s), 984 (m), 836 (m), 766 (s), 699 (s)
cm.sup.-1
[0271] EI-MS: (m/z) 60 (16), 91 (55), 120 (28), 152 (39), 182
(100), 225 (3), 327 (3), 404 (26) [M.sup.+]
[0272] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.67-7.66 (4H,
m), 7.61 (2H, d, J=8.2 Hz), 7.48-7.28 (8H, m), 5.70 (1H, s), 4.64
(1H, d, J=3.8 Hz), 4.34 (1H, d, J=9.5 Hz), 4.26 (1H, dd, J=10.3,
4.8 Hz), 4.24 (1H, d, J=4.8 Hz), 3.82-3.76 (2H, m), 3.65 (1H, dd,
app. t, J=9.2 Hz), 3.60 (1H, ddd, app. dt, J=9.5, 4.6 Hz), 3.54
(1H, ddd, app. dt, J=9.1, 4.9 Hz)
[0273] APT-NMR (125 MHz, CDCl.sub.3): .delta. 142.2 (C), 141.4 (C),
140.7 (C), 138.3 (C), 129.7 (CH), 128.7 (CH), 128.6 (CH), 128.5
(CH), 128.3 (CH), 127.8 (CH), 127.8 (CH), 127.3 (CH), 101.9 (CH),
83.6 (CH), 82.5 (CH), 76.9 (CH), 76.0 (CH), 71.7 (CH), 69.4
(CH.sub.2)
[0274] Synthesis of compound A7
[0275] To a solution of Compound 7.1 (150 mg, 0.371 mmol) in dry
DMF (5 ml), cooled to 0.degree. C., was added sodium hydride (60 mg
of a 60% dispersion, 4 eq). The reaction mixture was stirred at
0.degree. C. under argon atmosphere for 30 minutes. Subsequently
methyl iodide (120 .mu.l, 5 eq) was added. The reaction mixture was
stirred at room temperature under argon-atmosphere for 16 hours.
The reaction mixture was poured out into water (50 ml), followed by
separation of layers, and extraction of the aqueous layer with
diethyl ether (3.times.50 ml). The combined organic layers were
washed with brine (50 ml), dried over magnesium sulfate and the
solvent was removed under reduced pressure. The residue was
purified by column chromatography (230-400 mesh silica,
pentane:dichloromethane:et- her 10:10:0.25), to yield 147 mg
Compound A7 as a white solid. (92%).
[0276] Formula: C.sub.27H.sub.28O.sub.5
[0277] Molecular weight: 432.51
[0278] R.sub.f: 0.24 (pentane:dichloromethane:ether 10:10:0.25)
[0279] Melting point: 111-112.degree. C.
[0280] [.alpha.].sub.D.sup.20=-29.5.degree.;
[.alpha.].sub.365.sup.20=-110- .1.degree. (c=1.00 in
chloroform)
[0281] IR (KBr): 2932 (m), 2885 (m), 2834 (m), 1376 (m), 1173 (m),
1142 (m), 1103 (s), 1044 (m), 1033 (m), 1008 (m), 962 (m), 828 (m),
765 (s), 749 (s), 699 (s) cm.sup.-1
[0282] EI-MS: (m/z) 45 (18), 88 (43), 121 (100), 134 (18), 182
(31), 225 (20), 432 (2) [M.sup.+]
[0283] .sup.1H-NMR (500 MHz, CDCl.sub.3): .delta. 7.78-7.68 (5H,
m), 7.58-7.41 (7H, m), 5.81 (1H, s), 4.39 (1H, d, J=9.7 Hz), 4.35
(1H, dd, J=10.2, 4.9 Hz), 3.88 (1H, dd, app. t, J=10.1 Hz), 3.83
(1H, dd, app. t, J=9.2 Hz), 3.71-3.67 (1H, m), 3.69 (3H, s), 3.60
(1H, dd, app. t, J=9.1 Hz), 3.26 (1H, dd, J=9.5, 8.6 Hz), 3.11 (3H,
s)
[0284] APT-NMR (125 MHz, CDCl.sub.3): .delta. 141.4 (C), 140.6 (C),
139.6 (C), 137.4 (C), 128.9 (CH), 128.0 (CH), 127.6 (CH), 127.5
(CH), 126.9 (CH), 126.8 (CH), 126.5 (CH), 100.8 (CH), 85.4 (CH),
84.6 (CH), 82.0 (CH), 81.9 (CH), 70.5 (CH), 69.6 (CH.sub.2), 59.9
(CH.sub.3), 59.8 (CH.sub.3)
EXAMPLE 2
Bioactivity of the Compounds of Formula A
[0285] The compounds were screened against various pathogenic
viruses and more specific the human cytomegalovirus (CMV). For
determination of the antiviral activity, expressed in IC.sub.50,
against 2 CMV-strains (Davis and AD-169), human embryonic lung
fibroblast (HEL) cells grown in 96-well microplates were infected
with 20 PFU virus/well. After 2 hours of incubation at 37.degree.
C., the infected cells were replenished with 0.1 ml of medium
containing serial dilutions of the test compound. On day 7 the
plaques were counted microscopically after staining the cells with
Giemsa's solution. The minimum antiviral concentration was
expressed as the dose required to inhibit virus-induced plaque
formation by 50%.
[0286] The results of the screening of the new compounds against
the human cytomegalovirus (CMV) are presented in Table 3.
3TABLE 3 Antiviral activity of the compounds
IC.sub.50(.mu.g/ml).sup.a CMV AD-169 David Compound Strain Strain
Compound A1 2.7 2.0 Compound A2 5.0 20.0 Compound A3 2.5/2.7
3.2/2.8 Compound A4 0.9/1.3 >2/1.0 Compound A5 0.5 0.5 Compound
A6 0.47 0.8 Compound A7 0.8 0.6 Ganciclovir 1.3/2.5 0.5
.sup.ainhibitory concentration required to reduce virus plaque
formation by 50%. Virus input was 100 plaque forming units
(PFU)
[0287] Each of the compounds of Formula A showed a significant
activity against CMV. Activity of these new molecules (IC.sub.50 in
.mu.g/ml) are in the same range as the activity of ganciclovir for
CMV screenings.
[0288] Although the invention has been described with respect to a
preferred embodiment thereof, it is to be also understood that it
is not to be so limited since changes and modifications can be made
therein which are within the full intended scope of this invention
as defined by the appended claims.
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