JPH08127590A - 3'-glucosyladenosine derivative - Google Patents

Abstract

PURPOSE: To obtain the subject new derivative useful as a synthetic intermediate, etc., for substances raising Ca<2+> concentration by acting on inositol 1,4,5- triphosphate receptor, by condensing a specific adenosine derivative with a glucose derivative. CONSTITUTION: This new 3'-glucosyladenosine derivative is expressed by formula I [R<1> and R<2> are each an acyl; R<3> and R<5> are each a (substituted)benzyl, a trialkylsilyl, an alkoxyalkyl, etc.; R<4> is a (substituted)trrphenylmethyl or trialkylsilyl; R<6> to R<8> are each an acyl] and is useful as an important intermediate, etc., in the synthesis of adenophostins having activity to increase an intracellular calcium ion concentration by acting on inositol 1,4,5-triphosphate (InsP3) receptor. The derivative is obtained by condensing an adenosine derivative expressed by formula II with a glucose derivative expressed by formula III (X is an eliminable group).

Description

Detailed Description of the Invention

[0001]

The present invention relates to inositol 1,4.
The present invention relates to a key intermediate in the synthesis of adenophostins having an activity of acting on a 5-trisphosphate (InsP ₃ ) receptor to increase intracellular calcium ion concentration.

[0002]

2. Description of the ^Related Art It is widely accepted that Ca ²⁺ is important as an intracellular signal transmitter for neurotransmission, muscle contraction, cell proliferation and differentiation. To mobilize Ca ²⁺ into this cell,
Inositol 1,4,5-Trisphosphate (Ins
P ₃ ) has an important role (Nature, 341, 197-
205 pages (1989)). That, InsP ₃ is produced in vivo from phospholipids of cell membrane (ibid 312, pp. 315-321 (1984), InsP ₃ thus generated bind to InsP ₃ receptors located on the endoplasmic reticulum (ibid 342 Volume, Pages 32-38 (19
As a result, the Ca ²⁺ concentration stored in the endoplasmic reticulum is increased (Vol. 342, pp. 87-89 (1989)).

[0003] The present inventors Takahashi et al., The compound having the activity of increasing the Ca ²⁺ concentration acts similarly to InsP ₃ receptors InsP _3, Penicillium the Adenohosuchin (Adenophostin) such formula (II) brevicompactum SAN
It was found from cultures of K 11991 and SANK 12177 strains [JP-A-5-194580, J. Antibiot. 47, 95.
-100 pages (1994)].

[0004]

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(In the formula, R represents a hydrogen atom is adenophostin A, and R represents an acetyl group is adenophostin B.) However, adenophostins are produced by the fermentation method described in JP-A-5-194580. Since it is difficult to mass-produce adenophosphine, the present inventors tried the total synthesis of adenophostins.

[0006] As shown in the above formula (II), adenophostins have a characteristic structure in which the 3'hydroxyl group of adenosine is glycosically bonded to glucose by α-coordinate, but until now, the 3'hydroxyl group of adenosine was Nothing is known about the chemical synthesis of compounds glycosylated by glucose. FW Lichtenthaler and colleagues 5'of ribonucleosides
Reported glycosylation of hydroxyl groups [Angew. Chem, 9
0, 819-821 (1978)], and Suzuki et al. Reported that the enzymatic glycosylation of adenosine preferentially yielded a compound in which the 5'hydroxyl group was glycosylated (vitamin, 44 Vol. 196-200 (1971), none of which is glycosylation of the 3'hydroxyl group of adenosine.

[0007]

DISCLOSURE OF THE INVENTION The present invention provides a novel 3'-hydroxyl-glycosylated adenosine derivative which is useful as an intermediate which enables the total synthesis of adenophostins.

[0008]

Means for Solving the Problems As a result of intensive research to solve the above-mentioned problems, the present inventors succeeded in synthesizing a compound of formula (I) in which the 3'hydroxyl group of adenosine was glycosylated, and As an intermediate, it enabled the total synthesis of adenophostins.

[0009]

[Chemical 4]

[In the formula, R ¹ and R ² are the same or different and each represents an acyl group, R ³ and R ⁵ are the same or different, and a benzyl group which may have a substituent, a trialkylsilyl group, Represents an alkoxyalkyl group or an alkoxyalkoxyalkyl group, R ⁴ represents a triphenylmethyl group or a trialkylsilyl group which may have a substituent,
R ⁶ , R ⁷ and R ⁸ are the same or different and represent an acyl group] In the above formula, the acyl group of R ¹ and R ² is an alkanoyl group such as acetyl, propionyl, butyryl, isobutyryl, pentanoyl and pivaloyl. Na C ₂ ~
C ₈ alkylcarbonyl group; halogenated alkylcarbonyl group such as chloroacetyl, dichloroacetyl, trichloroacetyl, trifluoroacetyl; alkoxyalkylcarbonyl group such as methoxyacetyl,
Halogenated alkoxycarbonyl groups such as 2,2,2-trichloroethoxycarbonyl, as well as aromatic acyl groups such as benzoyl, bromobenzoyl, chlorobenzoyl; trimethylbenzoyl, toluoyl; anisoyl, nitrobenzoyl, 2- (methoxycarbonyl) benzoyl. Examples thereof include a benzoyl group which may be substituted with halogen, lower alkyl, lower alkoxy, nitro or lower alkoxycarbonyl, a phthaloyl group represented by R ¹ and R ² together, and preferably acetyl and benzoyl. is there.

The benzyl group which may have a substituent for R ³ and R ⁵ is benzyl, 4-bromobenzyl,
4-chlorobenzyl, 4-phenylbenzyl, methoxybenzyl, dimethoxybenzyl, halogen such as nitrobenzyl, benzyl optionally substituted by phenyl, lower alkoxy or nitro; trialkylsilyl group includes trimethylsilyl and triethyl. Silyl, isopropyldimethylsilyl, t-butyldimethylsilyl,
Methyldi-t-butylsilyl, triisopropylsilyl and the like; as the alkoxyalkyl group, methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, butoxymethyl, t-butoxymethyl, benzyloxymethyl and the like; as the alkoxyalkoxyalkyl group Examples include 2-methoxyethoxymethyl and 2-ethoxyethoxymethyl. Preferred R ³ and R
⁵ is benzyl, 4-nitrobenzyl, 4-methoxybenzyl, 3,4-dimethoxybenzyl, t-butyldimethylsilyl, 2-methoxyethoxymethyl and methoxymethyl.

Examples of the triphenylmethyl group which may have a substituent of R ⁴ include triphenylmethyl, 4-methoxytriphenylmethyl, 4,4'-dimethoxytriphenylmethyl, 4,4 ', 4 "-. Lower alkoxy such as trimethoxytriphenylmethyl, 4-cyanotriphenylmethyl or a triphenylmethyl group optionally substituted by cyano; examples of the trialkylsilyl group include those exemplified above for R ³ and R ^5. Preferred R ⁴ is triphenylmethyl, 4-methoxytriphenylmethyl, 4,4′-dimethoxytriphenylmethyl and t-butyldimethylsilyl.

The acyl group of R ⁶ , R ⁷ and R ⁸ is
Examples thereof include those exemplified for R ¹ and R ² , and acetyl and benzoyl are preferable.

Preferred compounds of the formula (I) can be exemplified in Tables 1 to 5, wherein the corresponding R ¹ and R ² are acetyl and / or the corresponding R ⁶ , R ⁷ and R
Equally preferred are compounds where ⁸ is benzoyl. In the table,
The abbreviations R ^{1 to} R ^{8 in} the following Reference Examples represent the following groups.

Bz = benzoyl Ac = acetyl Bn = benzyl PNBn = 4-nitrobenzyl MPM = 4-methoxybenzyl DMPM = 3,4-dimethoxybenzyl TBDMS = t-butyldimethylsilyl MEM = 2-methoxyethoxymethyl MOM = methoxymethyl Tr = triphenylmethyl MMTr = 4-methoxytriphenylmethyl DMTr = 4,4′-dimethoxytriphenylmethyl

[0016]

[Table 1]

[0017]

[Table 2]

[0018]

[Table 3]

[0019]

[Table 4]

[0020]

[Table 5]

In the above table, the most preferred compounds as the intermediates of the present invention are those having the following compound numbers. 1, 2,
6, 7, 11, 12, 41, 42, 46, 47, 51,
52, 61, 62, 63, 67, 68, 69, 73, 7
4, 75, 85, 86, 87, 91, 92, 93, 9
7, 98, 99, 109, 121, 133, 134, 1
35, 136 and 137.

The compound of formula (I) is produced by condensing a glucose derivative (IV) with an adenosine derivative (III).

[0023]

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(In the formula, R ^{1 to} R ⁸ have the same meanings as described above, and X represents a leaving group.) Examples of the leaving group include an alkanoyloxy group, an imide ether group, an alkylthio group and a halogen atom. The following suitable reaction conditions can be selected and carried out depending on the types of these leaving groups.

When X is an alkanoyloxy group,
For example, acetyloxy group, chloroform, 1,2
In a halogenated hydrocarbon solvent such as dichloroethane,
In the presence of a Lewis acid such as FeCl ₃ as an activator,
The reaction is carried out at 20 to 50 ° C for 1 hour to 1 day.

When X is an imide ether group, it is, for example, a trichloroacetimide ether group or an N-methylacetimide ether group, which is tosylic acid or trimethylsilyl triflate as an activator in a solvent such as ether or nitromethane. -20 in the presence of an acid catalyst such as
React at -50 ° C for 1-24 hours.

When X is an alkylthio group, it is, for example, a methylthio group and is an ether; an activator in a halogenated hydrocarbon such as 1,2-dichloroethane, dichloromethane or an aromatic hydrocarbon solvent such as toluene. In the presence of methyl triflate, cupric bromide and Bu ₄ NF, or silver triflate, bromine and tetramethylurea,
The reaction is carried out at -20 to 50 ° C for 1 hour to 5 days.

When X is a fluorine atom, benzene,
Sn as an activator in a solvent such as acetonitrile
Cl ₂ , or AgBF ₄ and (cyclopentadienyl) ₂ Z
The reaction is carried out in the presence of rCl ₂ at −20 to 50 ° C. for 1 to 48 hours.

When X is a chlorine or bromine atom, HgBr ₂ , Ag ₂ CO ₃ , silver as an activator in a halogenated hydrocarbon such as dichloromethane or chloroform, a solvent such as DMF, ether or nitromethane. Triflate and Collidine, Et ₄ NBr, or AgClO ₄
And in the presence of collidine, further using molecular sieves at -50 to 100 ° C for 5 hours to 7 days, if necessary.

After completion of the reaction, the reaction solution was filtered through Celite, the filtrate was diluted with an inert organic solvent such as chloroform, washed with 0.01N HCl, 5% NaHCO ₃ water and saturated saline solution in that order, and sulfuric acid was added. The desired product can be obtained by drying over magnesium and distilling off the solvent.

If necessary, it can be purified by silica gel chromatography.

The above compound (III) is produced by the following methods A-1 to A4 depending on the types of R ³ and R ⁴ .

(Method A-1) Compound in which R ³ is a benzyl group which may have a substituent and R ⁴ is a triphenylmethyl group or a trialkylsilyl group which may have a substituent (III-1) is produced by the method represented by the following reaction formula.

[0034]

[Chemical 6]

In the starting compound (1-1),
The compound in which R ³ is benzyl is described in J. Org. Chem. 37 , 339.
8 (1972), R ³ is 4-methoxybenzyl, and Chem. Lett. 189 (1982), and R ³ is 3,4-.
The compound that is dimethoxybenzyl is Chem. Lett. 1005.
(1986).

The first step consists of 3 steps per se, and compound (1-1) is reacted with trimethylsilyl chloride in pyridine at -20 to 50 ° C for 30 minutes to 5 hours.
The reaction mixture is reacted with benzoyl chloride, acetyl chloride, or the like at -20 to 50 ° C for 30 minutes to 10 hours, and then a small amount of water is added to the reaction mixture to give -20
The reaction is carried out at -50 ° C for 5 minutes to 2 hours.

After completion of the reaction, the solvent was distilled off, an inert solvent such as methylene chloride was added to the residue, and 5% NaHCO ₃ was added.
The desired product can be obtained by sequentially washing with water and water, drying over magnesium sulfate, and distilling off the solvent.

If necessary, it can be purified by silica gel chromatography.

In the second step, trityl chloride, 4-methoxyphenyldiphenylmethyl chloride, di (4-methoxyphenyl) phenylmethyl chloride and t-butyldimethylsilyl are added to compound (1-2) in a solvent such as pyridine or DMF. Chloride and the like are reacted at 0 to 100 ° C. for 30 minutes to 7 days. Moreover, you may make an imidazole coexist as an activator as needed.

After completion of the reaction, a small amount of water is added and the solvent is distilled off. Add a solvent such as ethyl acetate to the residue and add 5% Na.
Wash sequentially with HCO ₃ water, dry over magnesium sulfate,
It can be carried out by distilling off the solvent.

If necessary, it can be purified by silica gel chromatography.

(Method A-2) The compound (III-2) in which R ³ is a trialkylsilyl group and R ⁴ is an optionally substituted triphenylmethyl group is represented by the following reaction formula. It is manufactured by the method shown.

[0043]

[Chemical 7]

The third and fourth steps are adenosine (1
-3) is used as a raw material and R ¹ , R ² and R ⁴ are introduced under the same reaction conditions as in the first step and the second step, respectively. In the fifth step, the compound (1-5 )
T-butyldimethylsilyl chloride, etc.
R ³ is introduced by reacting at 1 ° C. for 1 hour to 5 days. Also,
In the fifth step, imidazoles may coexist as an activator, if necessary.

(Method A-3) The compound (III-3) in which R ³ and R ⁴ are trialkylsilyl groups can be produced by the method shown in the following reaction formula.

[0046]

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In the sixth step, compound (1-4) is reacted with t-butyldimethylsilyl chloride or the like at -20 to 50 ° C. for 1 to 7 days in a solvent such as pyridine or DMF. Moreover, you may make an imidazole coexist as an activator as needed.

(Method A-4) A compound in which R ³ is an alkoxyalkyl group or an alkoxyalkoxyalkyl group and R ⁴ is a triphenylmethyl group or a trialkylsilyl group which may have a substituent (III- 4) is produced by the method represented by the following reaction formula.

[0049]

[Chemical 9]

In the seventh step, the compound (1-
4) to tetraisopropyldisiloxane dichloride-
The reaction is carried out at 20 to 50 ° C for 2 hours to 3 days.

In the eighth step, methoxymethyl chloride or methoxyethoxymethyl chloride or the like is added to compound (1-6) in the presence of a base such as diisopropylethylamine in a solvent such as dichloromethane at -20 to 50 ° C. for 5 hours to
After reacting for 5 days, R ³ is introduced.

In the ninth step, the compound (1-
7) Tetrabutylammonium fluoride or the like in -20 to
The reaction is carried out at 50 ° C for 10 minutes to 5 days.

In step 10, R ⁴ is introduced by reacting compound (1-8) under the same conditions as in step 2.

The compound of the formula (IV) which is the other raw material is X
It is produced by the following methods B-1 to 5 depending on the type of the leaving group.

(Method B-1) The compound (IV-1) in which X is an alkanoyloxy group is produced by the method shown by the following reaction formula.

[0056]

[Chemical 10]

The eleventh step is J. Am. Chem. S when the starting material is pentaacetyl glucose (2-1).
oc. 74 , 1498 (1952). First
In the first step, piperidine is allowed to act on compound (2-1) at 25 ± 5 ° C. for 5 minutes to 2 hours without using a solvent.

In the 12th step, benzyl bromide or chloride optionally having a substituent in compound (2-2), t
- butyldimethylsilyl chloride, methoxyethoxymethyl chloride or methoxymethyl chloride, etc., Ag ₂
The reaction is carried out in the presence of O and a base such as diisopropylethylamine at −20 to 50 ° C. for 10 hours to 10 days. In the 13th step, compound (2-3) is treated under reflux in a mixed solvent of acetone-water-acetic acid for 1 minute to 1 hour.

In the fourteenth step, compound (2-4) is added with acetic anhydride or the like at -20 to 50 ° C. for 1 hour to 2 in a pyridine solvent.
React for days.

(Method B-2) The compound (IV-2) in which X is an imide ether group is produced by the method shown by the following reaction formula.

[0061]

[Chemical 11]

In the fifteenth step, in a dichloromethane solvent,
In the presence of a base such as sodium hydride, the compound (2
-4) with trichloroacetonitrile or the like at -20 to 50 ° C
And react for 5 minutes to 10 hours.

(Method B-3) The compound (IV-3) in which X is an alkylthio group can be produced by the method shown by the following reaction formula.

[0064]

[Chemical 12]

The 16th step is carried out in the same manner as the 14th step using acetic anhydride.

[0066] Step 17 is chloroform, 1,2-halogenated hydrocarbon solvent such as dichloroethane, -20 to 50 ° C. The Bu ₃ SnSMe like compound (2-5)
And react for 1 to 24 hours.

(Method B-4) The compound (IV-4) in which X is a fluorine atom is produced by the method shown by the following reaction formula.

[0068]

[Chemical 13]

In the eighteenth step, compound (2-4) is reacted with Et ₂ NSF ₃ at −20 to 50 ° C. for 1 to 24 hours in a solvent such as THF and dichloromethane.

(Method B-5) The compound (IV-5) in which X is a chlorine or bromine atom is produced by the method represented by the following reaction formula.

[0071]

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In the 19th step, HB was added to compound (2-5) in a solvent such as dichloromethane, chloroform or acetic acid.
r, TiBr ₄ , HCl, TiCl ₄ etc. at 0-100 ° C
And react for 1 to 24 hours.

With the compound of the formula (I) of the present invention, adenophostin A can be synthesized, for example, by the method shown in the following reaction scheme. For details, see the reference example below.

[0074]

[Chemical 15]

[0075]

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[0076]

Example 1 (1) 2'-O- (4-methoxybenzyl) -N, N-
Dibenzoyladenosine (1) 2'-O- (4-methoxybenzyl) adenine [H. Tak
aku et al., Chem. Lett., 189 (1982)] 2.022 g
(5.22 mmol) was azeotropically distilled with pyridine, dried, and then dissolved in 50 ml of pyridine and stirred in an ice-water bath. Trimethylsilane chloride 3.39 ml (26.1 mmol)
Was added and stirred for 30 minutes, and then benzoyl chloride 3.1 was added.
3 ml (26.1 mmol) was added and the mixture was stirred at room temperature. After 2 hours, 5 ml of water was added while cooling in an ice water bath. After 20 minutes, the solvent was distilled off, the residue was dissolved in 200 ml of methylene chloride, washed twice with 200 ml of 5% NaHCO ₃ water, and then the organic layer was dried over MgSO ₄ . After distilling off the solvent,
The residue was applied to a 200 g (230-400 mesh) silica gel column and eluted with 1 to 3% methanol-methylene chloride to obtain 0.846 g of the desired product. The fraction containing the target compound that could not be isolated was applied again to a 60 g (230-400 mesh) silica gel column, and 1% methanol-
Elution with methylene chloride gave 0.9939 g of the desired product (1). Therefore, the total amount of the target product is 1.8399 g (59%)
I got

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.51 (s,
1H, H-8); 8.06 (s, 1H, H-2); 7.87 (d, J = 7.26Hz, 4H,
Bz); 7.52 (t, J = 7.26Hz, 2H, Bz); 7.38 (t, J = 7.26Hz,
4H, Bz); 6.98 (d, J = 8.58Hz, 2H, MPM); 6.76 (d, J = 8.58
Hz, 2H, MPM); 5.91 (d, J = 7.26Hz, 1H, H-1 '); 5.52 (d,
J = 11.21Hz, 1H, OH); 4.87 (dd, J = 7.26Hz, J = 4.62Hz,
1H, H-2 '); 4.52-4.32 (m, 4H, H-3', H-4 ', PhC H ₂ ); 3.
95-3.65 (m, 2H, H-5 '); 3.75 (s, 3H, C H ₃ O); 2.60 (brs,
1H, OH). IR (CHCl ₃ ): 3300, 3000, 2925, 1705, 1599, 1579 MS: m / z 596 ([M + H] ⁺ ) (2) 2′-O- (4-methoxybenzyl) -5'-O
-(4-Methoxyphenyldiphenylmethyl) -N, N
-Dibenzoyladenosine (2) Compound (1) 0.9939 g (1.669 mmol) was azeotroped with pyridine and dried, and then dissolved in 20 ml of pyridine and 0.773 g (2.5 mmol) of 4-methoxyphenyldiphenylmethyl chloride. Was added and the mixture was stirred at room temperature. 6
After 6 hours, 1 ml of water was added to stop the reaction, and the solvent was distilled off. The residue was dissolved in 100 ml of ethyl acetate and 5% NaH
After washing twice with 100 ml of CO ₃ water, it was dried over MgSO ₄ . After distilling off the solvent, 100 g (70-230
applied to a silica gel column of (mesh) and eluted with hexane-ethyl acetate (1: 1, v / v) to obtain 1.3206 g (91%) of the desired product (2).

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.55 (s,
1H, H-8); 8.14 (s, 1H, H-2); 7.88-6.75 (m, 28H, Ar);
6.13 (d, J = 4.62Hz, 1H, H-1 '); 4.75-4.59 (m, 3H, H-
2 ', PhC H ₂ ); 4.35-4.30 (m, 1H, H-3'); 4.24-4.20 (m, 1
H, H-4 '); 3.77 (s, 3H, C H ₃ O); 3.76 (s, 3H, C H ₃ O); 3.
50-3.28 (m, 2H, H- 5 '); 2.61 (d, J = 5.28Hz, 1H, O H) IR (CHCl 3):. 3000, 2925, 1700, 1599, 1575, 1510 FAB-MS: m / z 868 ([M + H] ⁺ ) (3) 2'-O- (4-methoxybenzyl) -5'-O
-(4-Methoxyphenyldiphenylmethyl) -3'-
O- (3 ", 4", 6 "-tri-O-acetyl-2"-
O-benzyl-α-D-glucopyranosyl) -N, N-
Dibenzoyladenosine (3) Compound (2) 347 mg (0.4 mmol) and 3,4,6-tri-O-acetyl-2-O-benzyl-α-D-glucopyranosyl bromide [S. Brennan et al. ., J. Chem. So
c. (C) 1742 (1970)] 550.7 mg (1.2 mmol) was dissolved in 4 ml of chloroform, and the molecular sieve (4A) was dissolved.
2 g and 160 μl of 2,4,6-trimethylpyridine
(1.2 mmol) and 249 mg of AgClO ₄ (1.2 mmo
l) was added and the mixture was stirred at room temperature. After 24 hours, filter through Celite, dilute with 100 ml of chloroform, and add 0.01N
It was washed with 100 ml each of HCl, 5% aqueous NaHCO ₃ and saturated saline and then dried over MgSO ₄ . After distilling off the solvent, the residue was applied to a silica gel column of 100 g (70-230 mesh), and hexane-ethyl acetate (3:
2, v / v) to elute target product (3) 240.5 mg (48
%) Was obtained.

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.37 (s,
1H, H-8); 8.03 (s, 1H, H-2); 7.87-6.67 (m, 33H, Ar);
6.09 (d, J = 6.60Hz, 1H, H-1 '); 5.45 (t, J = 9.24Hz, 1
H, H-3 "); 5.37 (d, J = 3.30Hz, 1H, H-1"); 5.04-4.98
(m, 2H, H-2 ', H-4 "); 4.66-4.32 (m, 6H, H-3', H-4 ',
PhC H ₂ ); 4.15-4.08 (m, 2H, H-6 "); 3.89 (m, 1H, H-5");
3.77 (s, 3H, C H ₃ O); 3.72 (s, 3H, C H ₃ O); 3.62-3.57 (m,
1H, H-2 "); 3.57-3.27 (m, 2H, H-5 '); 2.01 (m, 9H, A
c). IR (KBr): 3062, 3032, 2937, 1752, 1706, 1600, 1576 FAB-MS: m / z 1246 ((M + H) ⁺ )

[0080]

Example 2 (1) 5.34 g (20 mmol) of N, N-dibenzoyladenosine (5) adenosine (4) was azeotropically dried with pyridine and then dissolved in 200 ml of pyridine,
Stir in an ice water bath. Trimethylsilane chloride 15.6 ml
(120 mmol) was added and the mixture was stirred for 30 minutes, then 14.4 ml (120 mmol) of benzoyl chloride was added, and the ice water bath was removed and the mixture was stirred for 2 hours. After the reaction, the mixture was stirred again in an ice-water bath, 20 ml of water was added, and the mixture was stirred. After 30 minutes, the solvent was distilled off, the residue was dissolved in 200 ml of water, extracted four times with 100 ml of ethyl acetate each time, and dried over magnesium sulfate. After the solvent was distilled off, the residue was applied to a silica gel column of 400 g (70-230 mesh), and 3-6% methanol-
Elute with methylene chloride to obtain 7.84 g (8) of the desired product (5).
2%) was obtained.

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.60 (s,
1H, H-8); 8.18-7.32 (m, 11H, Ph, H-6); 5.89 (d, 1H, J
= 6.60Hz, H-1 '); 4.90-4.82 (m, 1H, H-2'); 4.34 (d, 1
H, J = 4.62Hz, H-3 '); 4.29 (s, 1H, H-4'); 3.95-3.65
(m, 2H, H-5 '). (2) 5'-O- (4-methoxyphenyldiphenylme
(CHIL ) -N, N-dibenzoyladenosine (6) Compound (5) 4.277 g (8.99 mmol) was dissolved in pyridine 90 ml, 4-methoxyphenyldiphenylmethyl chloride 4.167 g (13.49 mmol) was added, and room temperature was added. It was stirred at. After 20 hours, 2 ml of water was added, the mixture was stirred for 10 minutes, and then the solvent was distilled off. The residue is ethyl acetate 300
The extract was dissolved in ₃ ml of water, washed with 300 ml of 5% NaHCO ₃ water three times, and dried with Na ₂ SO ₄ . After distilling off the solvent, the residue was applied to a silica gel column of 150 g (70-230 mesh), and hexane-ethyl acetate (1:
2, v / v), and 5.19 g (77) of the target product (6).
%) Was obtained.

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.60 (s,
1H, H-8); 8.32 (s, 1H, H-2); 7.87-6.73 (m, 24H, Ph);
6.01 (d, 1H, J = 5.69MHz, H-1 '); 5.10 (brs, 1H, OH);
4.83-4.78 (m, 1H, H-2 '); 4.41-4.35 (m, 2H, H-3', H-
4 '); 3.76 (s, 3H, CH ₃ O); 3.50-3.23 (m, 2H, H-5'); 2.
97 (brs, 1H, OH). IR (KBr): 3436, 3060, 2933, 1706, 1599, 1576, 1510,
1449 MS: m / z 748 ([M + H] ⁺ ) (3) 2'-O- (t-butyldimethylsilyl) -5 '
-O- (4-methoxyphenyldiphenylmethyl)-
2.905 g (3.885 mmol) of N, N-dibenzoyladenosine (7) compound (6) was dissolved in 8 ml of pyridine, 1.756 g (11.65 mmol) of t-butyldimethylsilyl chloride was added, and the mixture was stirred at room temperature. After 2 days, dilute with 300 ml of ethyl acetate and add 5% NaH.
After washing twice with 200 ml each of CO ₃ water, MgSO ₄
Dried in. After distilling off the solvent, 300 g (7
0-230 mesh) was applied to a silica gel column and eluted with hexane-ethyl acetate (7: 3, v / v) to obtain 0.8755 g of the desired product (7).

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.56 (s,
1H, H-8); 8.26 (s, 1H, H-2); 7.87-6.79 (m, 24H, Ph);
6.07 (d, 1H, J = 5.45Hz, H-1 '); 5.01-4.96 (m, 1H, H-
2 '); 4.35-4.25 (m, 2H, H-3', H-4 '); 3.77 (s, 3H, CH
₃ O); 3.56-3.33 (m, 2H, H-5 '); 2.66 (d, 1H, J = 3.85Hz,
OH); 0.82 (s, 9H, tBu); -0.03 (s, 3H, C H ₃ -Si); -0.21
(s, 3H, C H ₃ -Si). IR (KBr): 3538, 3061, 2952, 2930, 2858, 1708, 1600,
1576, 1510, 1449 MS: m / z 862 ([M + H] ⁺ ) (4) 2'-O- (t-butyldimethylsilyl) -5 '
-O- (4-methoxyphenyldiphenylmethyl) -3
′ -O- (3 ″, 4 ″, 6 ″ -tri-O-acetyl-
2 ″ -O-benzyl-α-D-glucopyranosyl)-
86 mg (0.1 mmol) of N, N-dibenzoyladenosine (8) compound (7) and 3,4,6-tri-O-acetyl-2-O-benzyl-α-D-glucopyranosyl bromide [S Brennan et al., J. Chem. Soc.
(C) 1742 (1970)] 138 mg (0.3 mmol) was dissolved in 1 ml of chloroform, and molecular sieve (4A) 0.5 was prepared.
g and 40 μl of 2,4,6-trimethylpyridine (0.3
mmol) and 62 mg (0.3 mmol) of AgClO ₄ were added, and the mixture was stirred at room temperature. After 5 days, filter through Celite, dilute with 50 ml of chloroform, add 0.01N HCl, 5% Na.
After washing with 50 ml each of HCO ₃ water and saturated saline, it was dried with MgSO ₄ . After the solvent was distilled off, the residue was applied to a silica gel column of 20 g (70-230 mesh), and hexane-ethyl acetate (2: 1, v /
Elution with v) gave 43.6 mg (35%) of the desired product (8).

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.48 (s,
1H, H-8); 8.11 (s, 1H, H-2); 7.86-6.78 (m, 29H, Ph);
6.01 (d, 1H, J = 6.33Hz, H-1 '); 5.48 (t, 1H, J = 9.68H
z, H-3 "); 5,46 (d, 1H, J = 3.77Hz, H-1"); 5.19-5.14
(m, 1H, H-2 '); 5.00 (t, 1H, J = 9.68Hz, H-4 "); 4.75
(d, 1H, J = 11.67Hz, PhC H ); 4.57 (d, 1H, J = 11.67Hz, P
hC H ); 4.45-4.30 (m, 2H, H-3 ', H-4'); 4.18-4.08 (m, 2
H, H-6 "); 3.95-3.85 (m, 1H, H-5"); 3.77 (s, 3H, CH
₃ O); 3.66-3.60 (m, 2H, H-2 ", H-5'a); 3.40-3.33 (m, 1
H, H-5'b); 2.01 (s, 3H, Ac); 1.99 (s, 3H, Ac); 1.98
(s, 3H, Ac); 0.72 (s, 9H, tBu); -0.13 (s, 3H, C H ₃ -S
i); -0.31 (s, 3H, C H ₃ -Si). IR (KBr): 3062, 3033, 2953, 2931, 2858, 1752, 1703,
1600, 1576, 1510, 1450 MS: m / z 1240 ((M + H) ⁺ )

[0085]

Reference Example 1 (1) 2'-O- (4-methoxybenzyl) -3'-O
-(3 ", 4", 6 "-tri-O-acetyl-2" -O
-Benzyl-α-D-glucopyranosyl) -N, N-di
Benzoyl adenosine (9 ) 240 mg (0.192) of the compound (3) obtained in Example 1
mmol) in 75 ml of chloroform and stirred in an ice-water bath. To this, 1.5 ml of trifluoroacetic acid was added, and after stirring for 10 minutes, 100 ml of 5% NaHCO ₃ water was added. The organic layer was washed with 100 ml of 5% aqueous NaHCO ₃ and then dried over MgSO ₄ . After distilling off the solvent, the residue was applied to a 60 g (70-230 mesh) silica gel column and eluted with ethyl acetate-hexane [(1: 1, v / v) and (2: 1, v / v)]. Target product (9) 129.3 mg
(69%) was obtained.

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.40 (s,
1H, H-8); 7.90 (s, 1H, H-2); 7.88-6.64 (m, 19H, Ar);
5.94 (d, J = 8.17Hz, 1H, H-1 '); 5.61-5.45 (m, 3H, H-
1 ", H-3", OH); 5.03-4.93 (m, 2H, H-2 ', H-4 "); 4.73-
3.62 (m, 12H, H-3 ', 4', 5 ', 2 ", 5", 6 ", PhC H ₂ ); 3.69
(s, 3H, C H ₃ O); 2.12 (s, 3H, Ac); 2.04 (s, 6H, Ac). IR (KBr): 2937, 1750, 1712, 1600, 1579, 1514 FAB-MS: m / z 974 ([M + H] ⁺ ) (2) 3'-O- (3 ", 4", 6 "-tri-O-acetate
Chill-2 ″ -O-benzyl-α-D-glucopyranos
) -N, N-Dibenzoyladenosine (10) Compound (9) 126 mg (0.129 mmol) was dissolved in methylene chloride 1.3 ml, water 0.14 ml and 2,3-dichloro-5,6-dicyano-1. , 4-benzoquinone 50mg
(0.22 mmol) was added and the mixture was stirred at room temperature. After 41 hours, the insoluble matter was filtered and washed with methylene chloride. The filtrate and the washing solution are combined, and 5% NaHCO ₃ water and saturated saline solution are added.
It was washed with each ml and dried over MgSO ₄ . After evaporating the solvent, the residue was applied to a silica gel column of 20 g (70-230 mesh), and ethyl acetate-hexane (2: 1,
v / v) to elute the desired product (10) 88.3 mg (80%)
I got

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 8.59 (s,
1H, H-8); 8.10 (s, 1H, H-2); 7.90-7.25 (m, 15H, Ar);
5.79 (d, J = 7.59Hz, 1H, H-1 '); 5.70 (d, J = 10.88Hz, 1
H, OH); 5.54 (t, J = 9.73Hz, 1H, H-3 "); 5.05-4.95 (m,
3H, H-2 ', 1 ", 4"); 4.72 (d, J = 11.55Hz, 1H, PhC H ₂ );
4.61 (d, J = 11.55Hz, 1H, PhC H ₂ ); 4.43 (d, J = 5.28Hz, 1
H, H-3 '); 4.32-3.70 (m, 8H, H-4', 5 ', 2 ", 5", 6 ", O
H); 2.08 (s, 3H, Ac); 2.07 (s, 3H, Ac); 2.05 (s, 3H, A
c). IR (KBr): 3468, 2935, 1751, 1713, 1600, 1578 FAB-MS: m / z 854 ([M + H] ⁺ ) (3) 5'-O- [di (4-methoxyphenyl) ) Pheny
Lumethyl] -3'-O- [6 "-O-di (4-methoxy)
Phenyl) phenylmethyl-2 "-O-benzyl-α-
D-glucopyranosyl] -N- [di (4-methoxyphen]
Nyl ) phenylmethyl] adenosine (11) Compound (10) (87.5 mg, 0.102 mmol) was dissolved in pyridine (1 ml), 29% aqueous ammonia (4 ml) was added, and the mixture was stirred at room temperature. After 26 hours, the solvent was distilled off, and azeotropic distillation with pyridine was performed three times. Dissolve the residue in 2 ml of pyridine,
Di (4-methoxyphenyl) phenylmethyl chloride 1
21 mg (0.359 mmol) was added and the mixture was stirred at room temperature. 9
After hours, the solvent was distilled off, the residue was dissolved in 100 ml of ethyl acetate and washed 3 times with 100 ml of 5% NaHCO ₃ water. After drying over Na ₂ SO ₄ , the solvent was distilled off and the residue was washed with 20
It was applied to a silica gel column of g (70-230 mesh) and eluted with 0.5-2% methanol-methylene chloride to obtain 99.4 mg (68%) of the desired product (11).

¹ H-NMR (270 MHz, 6 mg in 0.5 ml of CDCl ₃ ,
TMS) δ: 7.96 (s, 1H, H-8); 7.83 (s, 1H, H-2); 7.37-
6.66 (m, 44H, Ar); 6.06 (d, J = 6.93Hz, 1H, H-1 '); 4.8
2-3.10 (m, 35H, H-2 ', 3', 4 ', 5', 1 ", 2", 3 ", 4",
5 ", 6", C H ₃ O, OH, PhC H ₂ ). IR (KBr): 3412, 2929, 1607, 1509, 1465 FAB-MS: m / z 1426 ([M + H] ⁺ ) (4 ) 5'-O- [di (4-methoxyphenyl) phenyl
Lumethyl] -2'-O- (O, O'-xylylphospho
No) -3'-O- [2 "-O-benzyl-3", 4 "-
O-bis (O, O'-xylylphosphono) -6 "-O-
[Di (4-methoxyphenyl) phenylmethyl]]-N
-[Di (4-methoxyphenyl) phenylmethyl] ade
1H- in 58.5 mg (0.041 mmol) of nosin (12) compound (11)
20 mg (0.287 mmol) of tetrazole was added, and the mixture was azeotropically distilled with pyridine and dried. The residue was dissolved in 1 ml of acetonitrile, and 2-diethylamino-1,3,2-benzodioxaphosphane [Y. Watanabe et al., Tetrahedron was used.
Lett ,, 31 , 255 (1990)] was added and stirred at room temperature. Three
After 0 minutes, the reaction solution was cooled to -40 ° C, and acetonitrile 1
64 mg of metachloroperbenzoic acid dissolved in 0.3 ml (0.369
mmol) was added and the mixture was stirred for 20 minutes while returning to room temperature. After diluting the reaction solution with 100 ml of methylene chloride, 1
0% Na ₂ S ₂ O ₅ water, 0.01N HCl and 5% Na
It was washed with 100 ml of HCO ₃ water and dried over Na ₂ SO ₄ . The solvent was distilled off, and 10 g (70-230 mes) of the residue was added.
Apply to h) silica gel column, 0.5 ~ 1.5
Elution with% methanol-methylene chloride (12)
Obtained 75.9 mg (93%).

¹ H-NMR (270 MHz, CDCl ₃ , TMS) δ: 7.94 (s,
1H, H-8); 7.88 (s, 1H, H-2); 7.50-6.65 (m, 56H, Ar);
6.42 (d, J = 6.94Hz, 1H, H-1 '); 5.91 (m, 1H, H-2');
5.48-4.13 (m, 21H, H-3 ', 4', 5 ', 1 ", 3", 4 ", PhC
H ₂ ); 3.75-3.24 (m, 22H, H-2 ", 5", 6 ", C H ₃ O). IR (KBr): 2928, 1607, 1509, 1465 FAB-MS: m / z 1972 ( [M + H] ⁺ ) (5) Adenophostin A (13) Compound (12) 36.4 mg (0.0184 mmol) 80
% Acetic acid 3 ml, the mixture was allowed to stand for 1 hour, then the solvent was distilled off and azeotropically distilled several times with ethanol. 3 ml of the residue is ethanol
And dissolved in 1 ml of water, 3 mg of Pd-black was added, and the mixture was stirred under a hydrogen atmosphere (1 atm). After 24 hours, the mixture was filtered through Celite and washed with diluted aqueous ammonia and ethanol. The filtrate and the washing solution were combined, the solvent was distilled off, the residue was separated with water and 20 ml of ethyl acetate each time, the aqueous layer was concentrated, and then freeze-dried to obtain 12.2 mg (92%) of adenophostin A (13). It was

¹ H-NMR (270 MHz, D ₂ O, DOH as internal standard
(4.7 ppm)) δ: 8.22 (s, 1H, H-8);
8.11 (s, 1H, H-2); 6.19 (d,
J = 6.4 Hz, 1H, H-1 ′); 5.27
(D, J = 3.3 Hz, 1H, H-1 ″);
5.18 (m, 1H, H-2 '); 4.55
(M, 1H, H-3 ′); 4.43-4.32
(M, 2H, H-4 ', H-3 "); 3.96
-3.60 (m, 7H, H-5 ', 2 ",
4 ", 5", 6 "). IR (KBr): 3174, 1656, 140
2, 1044, 939 FAB-MS: m / z 668 ([M-H] ^- );
m / z 670 ([M + H] ⁺ )

 ─────────────────────────────────────────────────── ─── Continuation of front page (72) Inventor, Waiko, 1-2-2, Hiromachi, Shinagawa-ku, Tokyo Sankyo stock company (72) Hideji Takahashi, 1-258, Hiromachi, Shinagawa-ku, Tokyo Sankyo stock company

Claims (3)

[Claims] 1. An adenophostin intermediate compound represented by the following structural formula (I): Embedded image (In the formula, R ¹ and R ² are the same or different and represent an acyl group, and R ³ and R ⁵ are the same or different, and may have a benzyl group, a trialkylsilyl group, an alkoxyalkyl group. Or an alkoxyalkoxyalkyl group, R ⁴ represents an optionally substituted triphenylmethyl group or a trialkylsilyl group, and R ⁶ , R ⁷ and R ⁸ are the same or different and represent an acyl group) 2. R ¹ and R ² represent a benzoyl group or an acetyl group, and R ³ and R ⁵ are the same or different and each represents a benzyl group, a 4-nitrobenzyl group, a 4-methoxybenzyl group,
3,4-dimethoxybenzyl group, t-butyldimethylsilyl group, methoxymethyl group or 2-methoxyethoxymethyl group, R ⁴ is triphenylmethyl group, 4-methoxytriphenylmethyl group, 4,4′-dimethoxy The compound according to claim 1, wherein the compound represents a triphenylmethyl group or a t-butyldimethylsilyl group, and R ⁶ , R ⁷ and R ⁸ represent a benzoyl group or an acetyl group. 3. A process for producing an adenophostin intermediate compound (I) according to claim 1, wherein the adenosine derivative represented by the formula (III) is condensed with the glucose derivative represented by the formula (IV). Embedded image (In the formula, R ¹ and R ² are the same or different and represent an acyl group, and R ³ and R ⁵ are the same or different, and may have a benzyl group, a trialkylsilyl group, an alkoxyalkyl group. Or an alkoxyalkoxyalkyl group, R ⁴ represents an optionally substituted triphenylmethyl group or a trialkylsilyl group, R ⁶ , R ⁷ and R ⁸ are the same or different and represent an acyl group, X represents a leaving group)