JPS61229897A - Novel nucleoside compound - Google Patents

Abstract

NEW MATERIAL:The compound of formula I. EXAMPLE:2-Amino-3, 4-dihydro-5-formyl-7-beta-D-ribofuranosyl-7H-pyrrolo[2, 3-d]pyr imidin-4-one. USE:Antitumor agent. PREPARATION:The 6-halogen of the compound of formula II (R1 is alkoxy-alkyl; R2 is acyl; R3 and R4 are acyl, alkylidene or aralkyl; R5 is acyl or aralkyl; X is halogen) is reduced in the presence of a catalyst such as palladium-carbon. The reduction product is treated with an oxidizing agent such as active manganese dioxide in a solvent such as acetonitrile to oxidize the 5-hydroxymethyl to aldehyde and is deprotected to obtain the objective compound.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a novel nucleoside compound, a pharmaceutically acceptable salt thereof, a method for producing the same, and a pharmaceutical composition containing the compound as an active ingredient.

(Prior Art) Treatment techniques for various diseases are rapidly progressing, but malignant tumors are still in the early stages of research. the current,
Nucleoside compounds are known as thionosin and antitumor agents.
Only a few types, such as cytarabine, are used as pharmaceuticals, and research is being conducted to obtain more effective antitumor substances.

For example, as one of the nucleoside antitumor substances, cadeguomycin (2-amino-3,4-dihydro-4-
Oxo-7-β-D-ribofuranosyl-7H-pyrrolo(
2,3-d)pyrimidine-5-carboxylic acid) was disclosed in Japanese Patent Application Publication No. 5
It is disclosed in Japanese Patent No. 8-92693.

Cadeguomycin is produced in nature in very small amounts, so a method for mass synthesis is desired.
Kondo et al., Tetrahe
drone Letters+ 243647 (
An example of a synthetic method is disclosed in J. 1983).

(Problems to be Solved by the Invention) An object of the present invention is to provide a novel nucleoside compound, a pharmaceutically acceptable salt thereof, a method for producing the same, and a pharmaceutical composition containing the compound as an active ingredient. be.

(Means for Solving the Problems) While researching nucleoside compounds, their antitumor properties, and synthesis methods, the present inventors found that the nucleoside compound of the present invention, in which the carboxyl group at the 5-position of cadeguomycin was replaced with an aldehyde group, was discovered. The present inventors have discovered that the present invention has antitumor effects and is useful as an antitumor agent, and have completed the present invention.

The compound of the present invention is a novel nucleoside compound represented by the following general formula (1).

(In the formula, the wavy line indicates that the nucleoside compound of the present invention includes a bofuranose or arabinofuranose fraction.) The nucleoside compound of the present invention includes a pharmaceutically acceptable salt of the compound represented by the general formula (r), and includes, for example, ,
Examples include acid addition salts with inorganic acids such as hydrochloric acid and sulfuric acid, and acid addition salts with organic acids such as acetic acid and citric acid.

Next, a method for producing the compound of the present invention will be described.

For example, the compound of the present invention can be produced using a compound represented by the following general formula (II) as a starting material.

(In the formula, R1 is an alkoxyalkyl group, R2 is an acyl group, R3 and R4 are an acyl group, alkylidene group, or aralkyl group, R5 is an acyl group or an aralkyl group, and X is a halogen. Specific examples of the above general formula (II) (which represents a furanose group) include R1 being an alkoxyalkyl group such as a methoxymethyl group, R2 being an acyl group such as an acetyl or benzoyl group, and R3 and R4 being a general Any protecting group used in nucleoside chemistry, for example, an acyl group such as acetyl, benzoyl group, an aralkyl group such as trityl, benzyl group, or an alkylidene group such as isopropylidene, ethylidene group, R5 is also an acetyl group. , an acyl group such as a benzoyl group, an aralkyl group such as a trityl or benzyl group, and X may be a halogen such as chlorine or bromine.

Among the compounds represented by the general formula (rI), for example, 6-
Bromo-2-diacetylamino-3,4-dihydro-5
-Hydroxymethyl-3-methoxymethyl-7-(5-
0-acetyl-2,3-0-isopropylidene-β-D
-ribofuranosyl)-7H-pyrrolo[2゜3-d]pyrimidin-4-one is used as an intermediate in the synthesis of cadeguomycin rT, Kondo et al, + Tetrah
edronLetters, 24 3647 (1
983) can be easily obtained by known methods, as disclosed in J.

This manufacturing method can be performed as follows.

-M The halogen group at the 6-position of the compound represented by the formula (■) is reduced under a catalyst such as palladium-carbon.Then, a suitable solvent that does not inhibit the reaction, such as activated manganese dioxide in acetonitrile, is used. Treatment with an oxidizing agent oxidizes the 5-position hydroxylmethyl group and converts it into an aldehyde group. Finally, for example, after hydrolyzing the acetyl group by the action of aqueous ammonia in methanol, -a The nucleoside compound of the present invention represented by formula (1) can be obtained.

(2) The compound of the present invention can also be produced using cadeguomycin represented by the following formula (III) or its arabinose derivative as a starting material.

(In the formula, the wavy line represents the inclusion of bofuranose or arabinofuranose.) In this production method, first, cadeguomycin or its arabinose compound represented by the above formula (I [[)] is dissolved in methanol under a hydrochloric acid catalyst. The carboxyl group at the 5-position is esterified by heating under reflux, and then the alcohol is obtained by reacting with lithium borohydride under reflux in tetrahydrofuran. Finally, the 5-position methoxymethyl group is oxidized with activated manganese dioxide in dimethyl sulfoxide to obtain the compound of the present invention represented by general formula (1).

The obtained compound of the present invention can be obtained by distillation, chromatography,
Purify by normal means such as recrystallization, measure melting point, IR,
NMR, UV, -? Identification was done based on soot vector, optical rotation, etc.

(Example) An example of a method for producing the compound of the present invention is shown below.

Example 1゜(1) 6-bromo-2-diacetylamino-3,4-dihydro-5-hydroxymethyl-3-methoxymethyl-
7-(5-0-acetyl-2,3-0-isopropylidene-β-D-ribofuranosyl)-7H-pyrrolo(2,3
-d) To 545 g of pyrimidin-4-one were added 300 μm of 10% palladium-carbon, 300 μm of potassium acetate, and 20 μm of methanol/water, and the mixture was replaced with hydrogen gas and stirred at room temperature for 30 minutes. After removing the catalyst by filtration and washing, the filtrate was concentrated, water was added to the residue, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated sodium chloride, dried over anhydrous sodium sulfate, and then the solvent was distilled off. The obtained crude product was purified with a silica gel column and recrystallized from n-hexane to give 2-diacetylamino-3,4-dihydro-5-hydroxymethyl-3-methoxymethyl-7-(5-0- Acetyl-2,3-0-
Isopropylidene-β-D-ribofuranosyl)-7H-
390 quarts of white crystals of pyrrolo(2,3-d)pyrimidin-4-one were obtained.

(Yield 82%) Melting point: 57-59°C UV (MeOR): λmaX = 300 nt
a (ε deprivation 8770) λwax -265ns (
t-5414) I R (KBr): 3450.17
40.1690.1675.1572゜1372, 12
24.1090 cm-'NMR(CDC13)
: δ=1.35 (s, 3H), 1.60 (g, 3
B).

2.11 (s, 3H), 2.37 (s, 3H), 2
．． 38 (s, 311).

3.42 (s, 38), 4.20 (dd, IH, Jl
-6,6Hz.

Jz-12,71(z) I4-33 (dd, 1)
1, Jt 1=4-211z.

Jz-12-7Hz) 14.36(ddd, IHIJl
-3,7+1z.

J2114.2Hz, J3J, 6Hz) + 4.3-4
．． 5 (br, s.

IH), 4.75 (br, s, 2H), 4.80 (
dd, IH.

Jl-3,7To, Jz-6,3Hz), 4.93(
dd, IH.

Jl-2,9Hz, Jz-6,3Hz), 5.35
(s, 2H).

6.18 (d, IH, J=2.9Hz), 6.94 (
s, IH) [α) 12--47.9' (C-0,16,
CHCl3) (2) Product 39 obtained in (11) above
After dissolving 0.0 cm in 25 cm of acetonitrile, 3 g of activated manganese dioxide was added while stirring at room temperature. After stirring for 1 hour, it was filtered and washed with acetone, and the filtrate was concentrated. 15 volumes of methanol and 15 s of 28% aqueous ammonia were added to the residue, and after stirring at room temperature for 15 hours, the mixture was concentrated under reduced pressure. The obtained crude product was purified with a silica gel column and recrystallized from n-hexane to give 2-amino-3,4-dihydro-5-formyl-3-methoxymethyl-7-(2,3-0-isopropylene). Yellow crystals of lydene-β-D-ribofuranosyl)-7H-pyrrolo(2,3-d)pyrimidin-4-one 2
I got 08■. (Yield 71%) Melting point: 89-91°C UV (MeOH): λ-ax -317n+m
(ε, 6663)λwax −290ni+
Ct=5149)λa+ax -249ns
(t-21303)IR(KBr): 3440
．． 1695.1662.1630.1530°1205
, 1153.1077 tx-'N M R (CDCl
2): δ=L37(s, 3F+), 1.61(
s, 38).

3.44 (s, 3H), 3.77 (d, 18.J-1
2.2Hz).

3.95 (dd, IHIJl-2,011z, J2=1
2.2Hz).

4.43 (dd, IHlJl-2,01iz, Jz-2
,0Ilz).

5-02 (dd, IH9Jt-2,0Hz, Jz-5-
911z).

5.13 (dd, IHIJl-4, 4) 1z, Jz-5
, 911z).

5.50 and 5.61 (ABquartet, 2
H.

J=10.8H2), 5.60(br, s, 211)
.

5-75(d+IH1J=4-4FIz)-7,49(
s, 1ll).

10.25(s, 18) (a)” --77,4°(C=0.11.CHC
l)D″3 FARMass: mHz 395 (M+
1) (3) Trifluoroacetic acid/water (2/1) 15- was added to 200 μ of the product obtained in (2) above, and heated to 70°C.
The mixture was stirred for 1 hour.

After concentrating under reduced pressure, dissolve the residual fragrance in water and create Amber Ray) IR-
45 column and eluted with water/acetonitrile.

The eluate was concentrated to dryness to give 2-amino-3,4-dihydro-5
-Formyl-7-β-D-ribofuranosyl-7H-pyrrolo(2,3-d)pyrimidin-4-one (Compound 1) (133 ml) of yellow crystals was obtained. (Yield 85%) Melting point:
205-213 °C (decomposition) UV (H,,O)
: λ5rax -311nm (ε=745
2) λwax -245nm (ε-19110
)IR(KBr): 3410.3320.170
1.1685.1635°1598, 1406.118
5.1140°1045 cs-'NMR(DzOl t-BuOH-1,23ppm
): δ-3,8L (dd, IH, J1=4..2Hz,
J2=12.711z).

3.89 (dd, 18. Jl -3.1Hz,
J2-12.7Hz).

4.19 (dddtlH, J1=3.1Hz, J2=4
．． 2Hz.

J3=4.61(zL 4.35(dd, 11(, J1
=4.6Hz.

J2=5.1Hz)+4.57(dd,LH,J1=
5.1Hz.

J2=5.4Hz)+5.98(d+IH1J=5.
4FIz).

7.92 (s, 1), 9.68 (s, LH) [α).

=-47,7'(C,0,10,+(20)[α]ru 111.6°(C・0.10.H2O)FARMas
s: mHz 311 (M+1) Example 2゜6-bromo-2-diacetylamino-3,4-dihydro-5-hydroxymethyl-3-methoxymethyl-7-(
2,3,5-)di-0-acetyl-β-D-arabinofuranosyl)-7H-pyrrolo(2,3-d)pyrimidine-
2-amino-3, which is the arabinose form of Compound 1, was prepared in the same manner as in Example 1 using 290 μl of 4-one as a starting material.
4-dihydro-5-formyl-7-β-D-arabinofuranosyl-7H-pyrrolo(2,3-d)pyrimidine-4
-one (compound 2) 30 μm was obtained.

Melting point: 203-210 ℃ (decomposition) UV
(H2O): λwax = 311 rv (
g = 10845) λwax = 247 n-
(t-26895) IR (Br): 3400.3
330.3200.16B5.1645°1612, 1
552.1498.1380゜1067 cs-' NMR (mouth 20.t-BuO) I-1,23ppl
l): δ-3,8L (dd, IH, J1=5.1Hz,
J2”12.5+1z)+3.93(dd, 1)1.
Jl-3,211z, J2-12.511z).

4.01 (ddd, IH, J1=3.2Hz, J2
-5.1Hz.

J3=5.6Hz), 4.27(t, lIl, J=5
．． 6Hz).

4.48 (t, 18.J=5.6Hz), 6.32
(d, LH.

J=5.6Hz), 7.97(s, 1M), 9
．． 71 (s, IH) [α] Sweet +35.2° (C=0.
1. H2O) [α bottle +36.0″ (C=0.1.
H2O) Example 3 (1) Anhydrous methanol and concentrated sulfuric acid were added to 20 ml of cadeguomycin, and the mixture was refluxed on an oil bath for 4 hours. It was concentrated under reduced pressure, and the residual aroma was dissolved in water, passed through an Amberlite IR-45 column, and eluted with methanol/water. The eluate was concentrated to dryness, and methyl 2-amino-3,4-dihydro-4-,t-1
-so7-β-D-ribofuranosyl-7H-pyrrolo(2
, 3-d) Pale yellow crystals of pyrimidine-5-carboxylate (19) were obtained. (Yield 91%) Melting point: 2
40-242℃ (decomposition) UV (H2O):
λmax -296nm (8,8840)λwa
x = 270 nm (εII 7782) λ-a
X” 232 nm (ε= 19253
)IR(KBr): 3400.1711.164
5.1600.1550°1512, 1442.139
B, 1300.1212°1092 Rho 1 N M R (020, t-BuOH=1.23 ppl
l): δ-3,8L(s,38), 3.65-3
．． 96 (m, 21+).

4.16 (q, IH, J=3.2Hz), 4.34
(t, IH.

J=4.6Hz), 4.55(t, LH, J-5,
0Hz).

5.94 (d, IH, J=5.1Hz), 7.72
(s, IH) [α)0=-49,2(C=0.12.
H2O) (2) Dry tetrahydrofuran (3) and lithium borohydride (10) were added to 15 quartz of the product obtained in (1) above, and after purging with nitrogen, the mixture was refluxed on an oil bath for 46 hours. After tRKfi under reduced pressure, the residual aroma was dissolved in water and purified by preparative high performance liquid chromatography to obtain 2-amino-3,4-dihydro-5-hydroxymethyl-7-β-D-ribofuranosyl-7H-pyrrolo(2,3- d) 6 cm of white crystals of pyrimidin-4-one were obtained. (Yield 44%) Melting point: 230-235°C (Decomposition) UV
(H2O): λ-ax -283ns+
(ε= 6771)λwax -261am
Ca= 10142)λsgax -221nm
(εII 17823)r R(KBr): 3
460.3370.1652.1632.1610゜1
450, 1198.1130.1042.1014°8
60 clm-'NMR(DMSO-d6/D20=1/1.t-
BuOH=1.23 pPII+): δ=3.73(
dd, IH, J114. IHz, J2-12°311.

3.75 (dd, LH, J1=3.5112.J
2-12.3H2).

4.08 (ddd, IH, J1=3.0H2, J2=
3.5H2°J3=4. IH2), 4.25(dd, 1
8. J1=3.0Hz.

J2 = 5.5Hz), 4.51 (dd, I
H, J1□ 5-5Hz.

J2-6.8Hz), 4.68(s, 2H), 5.
91 (d, LH.

J-6,8Hz), 6.94(a, LH)(α)13-
-48.0″(C-0,06,nMso) (3) Above (
After dissolving the product 51Ilr obtained in 2) in dimethylformamide 2-, 50 μm of activated manganese dioxide was added and stirred at room temperature for 15 hours. After filtering through Celite and washing with water/methanol, the filtrate was concentrated.

The residue was purified by preparative high performance liquid chromatography and 2.
5N compound 1 was obtained. (Yield 50%) The melting point and IR of the compound obtained in Example 3 above.

NM)2. Physical property values such as UV, mass spectrum, and optical rotation were completely inferior to those of Compound 1 obtained in Example 1, so it was confirmed that the same substance was produced by both production methods.

(effect) The pharmacological actions of the compounds of the present invention will be described below.

(1) Growth inhibition effect on mouse lymphoma L5178Y cells Compound 1 and cadeguomycin were added to the culture medium of mouse lymphoma L5178Y cells at 37°C.
After culturing for 72 hours, the cell number was measured using an electric cell count (manufactured by Coulter), and the tumor cell growth inhibiting effect of the test compound was investigated. Mouse lymphoma L 5178 Y cells were cultured in Fisher's medium supplemented with 10% horse serum.

The results are shown in Figure 1.

(2) Effect of promoting [3H]thymidine uptake by 562 cells in human leukemia Compounds 1, 2 and cadeguomycin were added to 562 cells in human leukemia, and the mixture was incubated at 37°C under a flow of 5% carbon dioxide and 95% air. After culturing for an hour, [3H] thymidine was added and the mixture was further cultured for 6 hours. cells Ce1l-Ha
The effect of the test compound on promoting the uptake of [3H]thymidine in human leukemia 562 cells was investigated by collecting the radioactivity using a rvester and measuring the radioactivity taken up into the cells. Human leukemia 562 cells were cultured in PRMI-1640 medium supplemented with 10% fetal bovine serum.

The results are shown in Figure 2.

(Effect) As shown in FIG. 1, the compound of the present invention significantly inhibits the growth of mouse lymphoma L 5178 Y cells compared to cadeguomycin. The rc50 of the tumor cell growth inhibiting effect of Compound 1 is as low as LOpg/-, and the compound of the present invention exhibits antitumor activity in trace amounts.

Furthermore, as shown in FIG. 2, the compound of the present invention significantly promotes the uptake of [3H]thymidine by 562 cells in human leukemia. In particular, the ribose form of the compound of the present invention (compound l) is
Its effect is remarkable even at low concentrations, as it shows approximately 2.5 times more [3■] thymidine uptake promoting action than cadeguomycin at a concentration of 0.5 pg/bark.

As is clear from the above experimental results, the nucleoside compound of the present invention exhibits excellent antitumor effects and is useful as an antitumor agent. In addition, the compound of the present invention has a blood cell increasing effect,
It can also be expected to have an immune-enhancing effect by activating lymphocytes.

The compound of the present invention can be combined with a suitable pharmaceutical carrier or diluent to form a medicament, and can be formulated by any conventional method to form a solid, semisolid, liquid or gaseous form for oral or parenteral administration. It can be formulated into the following dosage forms.

In the formulation, the compounds of the present invention may be used in the form of their pharmaceutically acceptable salts, and the compounds of the present invention may be used alone or in appropriate combinations, and the compounds may be used alone or in appropriate combinations, and may be used in combination with other pharmaceutically active antitumor substances. It may also be used in combination with other ingredients.

As an oral preparation, it can be used as it is or with appropriate additives,
Binders such as crystalline cellulose, cellulose derivatives, gum arabic, corn starch, gelatin, corn starch, along with customary excipients such as lactose, mannitol, corn starch, potato starch, etc.
Disintegrants such as potato starch and sodium carboxymethyl cellulose, lubricants such as talc and magnesium stearate, other fillers, wetting agents, buffers, preservatives, fragrances, etc. are combined as appropriate to form tablets, powders, granules, or capsules. It can be used as an agent.

Furthermore, the compound of the present invention can be mixed with various bases, such as oily bases such as cacao butter, emulsion bases, water-soluble bases such as macrogol, hydrophilic bases, etc., to form suppositories. I can do it.

The injection may be a solution or suspension in an aqueous or non-aqueous solvent, such as distilled water for injection, physiological saline, Ringer's solution, vegetable oil, synthetic fatty acid glyceride, higher fatty acid ester, or propylene glycol.

In addition, depending on the site of tumor occurrence, it can be formulated into dosage forms other than those mentioned above that are most suitable for the treatment, such as inhalants, aerosols, eye drops, ointments, and bumps.

The desired dosage of the compound of the present invention varies depending on the subject to be administered, the dosage form, the method of administration, the administration period, etc., but in order to obtain the desired effect, it is generally necessary for an adult to administer 1 to 3,000 doses of the compound of the present invention per day. (1), preferably 5 to 1,500 (2), can be administered, and one to several units of a unit preparation containing an appropriate amount of the compound of the present invention can be administered per day.

(Example) Examples of formulations of pharmaceutical compositions containing the compound of the present invention as an active ingredient are shown below, but the present invention is not limited thereto.

Prescription example 1. (Tablet) Ingredients per tablet (■) Compound of the present invention 50 milk W
180 corn starch
40 Magnesium stearate 10 total 280■ Prescription example 2. (Capsules) Ingredients Per 1 capsule (■) Compound of the present invention 50 lactose
250 total 300■ Prescription example 3. (Injection) Ingredients per ampule (■) Compound of the present invention 10 Sodium chloride
Appropriate amount: 1- Prescription example 4. (Suppositories) Compound of the present invention 100 cocoa butter
1900 (6891) Patent attorney Murayama Sabube 4 guests lunch 6) - Ri l) Orange 9 Kui Yuma 1 - Hata ≦ Procedure (Ho Seisho (Method) (2) No suffix 1985 7
Month 2nd

Claims (3)

[Claims] (1) A novel nucleoside compound represented by general formula (I) and a pharmaceutically acceptable salt thereof. ▲There are mathematical formulas, chemical formulas, tables, etc.▼(I) (2) General formula (II) (wherein, R_1 is an alkoxyalkyl group, R_2 is an acyl group, R_3, R_4 are an acyl group, alkylidene group, aralkyl group, R_5 is an acyl group, an aralkyl group,
represents a halogen) and converts it into an aldehyde group by oxidizing the hydroxymethyl group of the compound, and then removing the protective group to form the general formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (I) A method for producing a novel nucleoside compound. (3) General formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼An antitumor agent containing a novel nucleoside compound represented by (I) or a pharmaceutically acceptable salt thereof as an active ingredient.