JP3067233B2 - Adenosine derivatives and uses thereof - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel adenosine derivative and its use.

[0002]

2. Description of the Related Art Most of the existing anticancer drugs have an action of inhibiting DNA synthesis or cell division, but they have an equivalent action on normal cells. Cancer treatment is established by giving more damage to cancer cells by utilizing the difference that cell division is slightly faster in normal cancer cells and slow in normal cells. The damage that normal cells suffer is expressed as a side effect, and the extent to which a living body can withstand the side effect is an important point in cancer treatment.

[0003] As is clear from the above, the original cancer treatment should be based on the biology, biochemistry, and the like of cancer cells, but it is not actually linked to such cancer treatment methods.

[0004] By the way, three causes of cancer, carcinogens, radiation and viruses, have long been said. Among them, the genetic information of cancer viruses has revealed that cells become cancerous, and the term oncogene (oncogene) was coined. Later, it was hypothesized that oncogenes would also be present in normal cells, which would be switched on at some point, causing the cells to become cancerous. It has evolved over time, and it will be appreciated by those skilled in the art that the outline was correct today.

On the other hand, in the genome of higher animals, there are 50 or more proto-oncogenes that can be oncogenes, and they play important physiological functions for the growth and differentiation of normal cells.
Therefore, the possibility of controlling cell proliferation and cancer at the gene level or gene product level has been developed.

An object of the present invention is to provide a therapeutic agent for cancer which specifically inhibits the expression of an oncogene and suppresses it.

[0007] By the way, mouse mammary carcinoma cells in which the expression of the inserted mouse mammary tumor virus (MMTV) gene is regulated by corticoids are used, and the depolyADP-ribose reaction with chromatin protein is triggered for the MMTV gene expression. Has been found to be. That is, it is considered that the degradation of poly-ADP-ribose causes local changes in the chromatin structure at that portion, ultimately leading to the binding of RNA polymerase to the promoter and the promotion of transcription [Journal Biological. Chemistry, 258 , 15371 (1983)].

[0008] Under such circumstances, the inventor has found that poly-ADP
In anticipation of inhibiting ribose degradation, it is expected that oncogenes will not be activated, and poly (ADP-ribose) glycohydrolase, an enzyme involved in ADP-ribose degradation, is separated and purified from human placenta, A compound having an inhibitory effect on was studied. As a result, a strong inhibitory activity was found for some novel compounds. Furthermore, the present inventors have further studied and succeeded in creating a new compound that can be used as a drug having an anticancer effect based on poly (ADP-ribose) glycohydrolase inhibition and completed the present invention.

[0009]

That is, the gist of the present invention is as follows. General formula

Embedded image [Wherein, R ¹ is a hydrogen atom;

Embedded image Or A (where A is a carbonyl having a phenyl substituted with a plurality of groups selected from a hydroxyl group and a lower alkoxy), and R ^{2 to} R ⁶ are each a hydrogen atom or A (where A is As defined above). However, R ^{1 to} R ³ and R ^{2 to} R ⁶ do not simultaneously represent a hydrogen atom. ] The adenosine derivative represented by these.

A poly (ADP-ribose) glycohydrolase inhibitor comprising the adenosine derivative described above as an active ingredient.

A cancer immunotherapy comprising the above-mentioned adenosine derivative as an active ingredient.

In the present specification, the lower alkoxy represented by A preferably has 1 to 4 carbon atoms, and specifically includes methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert- Butoxy and the like are exemplified, and methoxy is particularly preferred.

[0013] As A, a compound in which phenyl and carbonyl are bonded and a compound in which phenyl and carbonyl are directly bonded through alkylene or alkenylene are particularly preferable. As alkylene, methylene, ethylene,
Examples thereof include those having 1 to 4 carbon atoms such as trimethylene and tetramethylene, and methylene and ethylene are particularly preferable.
Examples of alkenylene include those having 1 to 4 carbon atoms, and vinylene is particularly preferable.

A preferred example of A is a compound represented by the general formula:

Embedded image (In the formula, Z represents a direct bond, alkylene or alkenylene, and R ^{7 to} R ¹¹ each represent a hydrogen atom, a hydroxyl group or lower alkoxy. However, R ^{7 to} R ¹¹ represent 4 or 5 hydrogen atoms simultaneously. It is not shown.)

Particularly preferred specific examples of A include galloyl, 4-hydroxy-3-methoxybenzoyl, 4-hydroxy-3,5-dimethoxybenzoyl, 3,4,5
-Trimethoxybenzoyl, 4-hydroxy-3-methoxycinnamoyl, 4-hydroxy-3,5-dimethoxycinnamoyl, 3,4,5-trimethoxycinnamoyl, 3,4,5-trihydroxybenzylcarbonyl,
3,4,5-trihydroxyphenethylcarbonyl and the like are exemplified.

The method for producing the adenosine derivative (I) of the present invention includes, for example, the following method.

Embedded image (Wherein A is as defined above, R ^{1 ′} is a hydrogen atom or

Embedded image Represents)

The above reaction is carried out by a usual ester reaction.

Compounds (i) and (ii) as starting materials
Are both known compounds and can be easily obtained. The compound (i) is an alcohol, and the compound (ii) is a carboxylic acid.

[0019]

Operation and effect of the present invention Adenosine derivative (I) of the present invention
Are mammals including humans (humans, horses, dogs, mice,
It has poly (ADP-ribose) glycohydrolase activity against guinea pigs, rats, etc., and is useful as a poly (ADP-ribose) glycohydrolase inhibitor for the treatment and prevention of malignant tumors and viral infections.

[0020] The adenosine derivatives of the present invention may be administered by themselves or in the form of a pharmaceutical formulation with a pharmaceutically acceptable carrier. The formulations are prepared by methods known per se. Examples of the dosage form include tablets, capsules, powders, suppositories, and injections.

The adenosine derivative of the present invention is administered orally or parenterally.

The dose of the adenosine derivative of the present invention may vary depending on the age and weight of the patient, the severity of the condition to be treated, and the response to therapy. For example, in the case of oral administration, it is usually 0.1 to 100 mg / kg body weight. The extent is administered once or several times a day.

[0023]

EXAMPLES The present invention will be described below in more detail with reference to examples, but the present invention is not limited to these examples.

Example 1 Synthesis of 2 ', 3', 5'-tri-O-galloyladenosine Dimethylformamide (50 ml), gallic acid (10 g)
And the solution obtained from benzyl chloride (27 ml)
Dilute with ethyl acetate stirred at 0 ° C. overnight. The ethyl acetate layer was washed with water and saturated saline, and dried over magnesium sulfate. After evaporating the solvent under reduced pressure, ethanol (200 m
l) and 1.6N aqueous sodium hydroxide solution (50 ml)
And heated under reflux for 2 hours. After the reaction, about half of the ethanol was distilled off using an evaporator, and the obtained residue was evaporated to 0%.
The solution was cooled to 0 ° C and the solution was adjusted to pH 2 with 0.5N hydrochloric acid. At that time, the precipitated solid was collected by filtration, and dried to obtain the compound (1).
(5.6 g, 64%). Compound (7.0 g), thionyl chloride (40 ml) and dimethylformamide (1 m
l) was added. The resulting solution was heated to reflux overnight, after which excess thionyl chloride was distilled off under normal pressure and reduced pressure.
4,5-Tribenzyloxybenzoyl chloride was prepared.

Adenosine (500 mg), pyridine (10
ml), 3,4,5-tribenzyloxybenzoyl chloride (5.3 g) was added to the solution at room temperature.
Stir for days. After the reaction, the reaction solution was diluted with ethyl acetate, and the obtained ethyl acetate layer was washed with water, 1N hydrochloric acid and saturated saline, and dried over magnesium sulfate. After evaporating the solvent under reduced pressure, the residue was subjected to silica gel column chromatography [silica gel, solvent: hexane: ethyl acetate = 1/2, 2/3] to give N, N ', 2', 3 ', 5'-pentakis (3 , 4,5-
Tribenzyloxybenzoyl) adenosine (intermediate compound 1) (3.5 g, 76%) was obtained.

Intermediate compound 1 (1.0 g), benzoylhydrazine (0.55 g) and pyridine (5 ml) were added.
The resulting solution was heated under reflux for 4 hours, after which the solvent pyridine was distilled off under reduced pressure. The obtained residue was diluted with ethyl acetate, and the ethyl acetate layer was washed with brine and dried over magnesium sulfate. After evaporating the solvent under reduced pressure, the residue was subjected to silica gel column chromatography [silica gel, solvent: ethyl acetate: hexane = 1/2, 1/1, 2/1] to give 2 ′,
3 ', 5'-Tris-O- (3,4,5-tribenzyloxybenzoyl) adenosine (intermediate compound 2) (410
mg, 66%).

Palladium-black (2.4 g) was added to the intermediate compound 2 (2.4 g) and ethyl acetate / methanol (1/1: 100 ml), and the reaction was started under a hydrogen atmosphere. After stirring overnight at room temperature, the palladium-black was filtered off. The obtained filtrate is concentrated and subjected to liquid chromatography (column: asahipackODP-50 preparative column, solvent:
The target compound 2 ′, 3 ′, 5′-tri-O-galloyladenosine (0.75 g, 66%) was obtained.

¹ H-NMR (DMSO-d ₆ ) δ: 4.5
-4.9 (m, 3H), 5.9-6.0 (m, 1H),
6.1-6.2 (m, 1H), 6.43 (d, J = 5.
6 Hz), 6.88 (s, 2H), 6.99 (s, 2H)
H), 7.04 (s, 2H), 7.44 (brs, 2
H), 8.19 (s, 1H), 8.35 (s, 1H),
8.9-9.6 (m, 9H)

IR (KBr, cm ^-1 ): 3300, 17
00,1635,1600

Experimental Example 1 Poly (ADP-ribose) gly
Cohydrolase inhibition assay buffer (0.01% bovine serum albumin-10 mM mercaptoethanol-50 mM potassium
Phosphoric acid, pH 7.0), ³ H- (ADP-ribose)
_{n = 15} was added, and a test substance and a nucleus-derived, poly (ADP-ribose) glycohydrolase solution prepared from the human placenta were added to 27 μL to make a total volume of 30 μL.
For 1 hour. After that, DE81
After absorbing the reaction solution with filter paper, washing the filter paper with water, ethanol and acetone, drying the filter paper, measuring the unreacted substrate ³ H- (ADP-ribose) with a liquid scintillation counter, and testing the enzyme. The inhibitory effect of the substance was studied. As a result, the IC ₅₀ value of 2 ′, 3 ′, 5′-tri-0-galloyladenosine was 30 μg / ml.

Formulation Example 1: Tablets 10 g of the compound of the present invention No. 209 for direct hitting No. 209 (manufactured by Fuji Chemical Co., Ltd.) 110 g Magnesium aluminate metasilicate 20% Maize starch 30% Lactose 50% Crystalline cellulose 60 g CMC calcium 18 g Magnesium stearate 2 g

Both and are previously passed through a 100-mesh sieve. After each of these is dried to reduce the water content to a certain level, they are mixed at the above weight ratio using a mixer. The whole mixed powder was added and mixed for a short time (30 seconds), and the mixed powder was tabletted to give a tablet of 200 mg per tablet.

The tablet may be coated with a commonly used gastrosoluble film coating agent (eg, polyvinyl acetal diethylaminoacetate) or an edible coloring agent, if necessary.

Formulation Example 2: Capsule Compound of the present invention 50 g Lactose 930 g Magnesium stearate 20 g

The above components were weighed and mixed uniformly, and the mixed powder was filled into hard gelatin capsules in an amount of 200 mg each.

Formulation Example 3: Injection Compound of the present invention 5 mg Glucose 100 mg Physiological saline 10 mL

After filtering the above mixture through a membrane filter, sterilization filtration was performed again. The filtrate was aseptically dispensed into vials, filled with nitrogen gas, and sealed to obtain an intravenous injection. .

Continuation of the front page (56) References Annals of the New York Academy of Sciences, Vol. 284, p. 81-90 (1977) (58) Fields investigated (Int. Cl. ⁷ , DB name) C07H 19/167, 19/19 A61K 31/7076 A61P 35/00, 43/00 CA (STN) REGISTRY (STN)

Claims (3)

(57) [Claims] 1. A compound of the general formula [Wherein, R ¹ is a hydrogen atom, a general formula: Or a group represented by the general formula : (Wherein Z is a direct bond, alkylene or alkenylene
R ^{7 to} R ¹¹ each represent a hydrogen atom, a hydroxyl group or a low
Represents a lower alkoxy. However, R ⁷ ~R ¹¹ is four simultaneous to
Or 5 hydrogen atoms. )
A, and R ^{2 to} R ⁶ are each a hydrogen atom or A
(Where A is as defined above). However, R ¹ ~
R ³ and R ^{2 to} R ⁶ do not simultaneously represent a hydrogen atom. Adenosine derivative [5 '
(3,4,5-trimethoxybenzoyl) -arabinoff
Excluding lanosyl adenosine] . 2. A compound of the general formula [Wherein, R ¹ is a hydrogen atom, a general formula : Or a group represented by the general formula : (Wherein Z is a direct bond, alkylene or alkenylene
R ^{7 to} R ¹¹ each represent a hydrogen atom, a hydroxyl group or a low
Represents a lower alkoxy. However, R ^{7 to} R ¹¹ are four at the same time.
Or 5 hydrogen atoms. )
A, and R ^{2 to} R ⁶ are each a hydrogen atom or A
(Where A is as defined above). However, R ¹ ~
R ³ and R ^{2 to} R ⁶ simultaneously represent a hydrogen atom.
Absent. And a poly (ADP-ribose) glycohydrolase inhibitor comprising an adenosine derivative represented by the formula: 3. A compound of the general formula [Wherein, R ¹ is a hydrogen atom, a general formula : Or a group represented by the general formula : (Wherein Z is a direct bond, alkylene or alkenylene
R ^{7 to} R ¹¹ each represent a hydrogen atom, a hydroxyl group or a low
Represents a lower alkoxy. However, R ^{7 to} R ¹¹ are four at the same time.
Or 5 hydrogen atoms. )
A, and R ^{2 to} R ⁶ are each a hydrogen atom or A
(Where A is as defined above). However, R ¹ ~
R ³ and R ^{2 to} R ⁶ simultaneously represent a hydrogen atom.
Absent. ] A cancer immunotherapy comprising an adenosine derivative represented by the formula [1 ] as an active ingredient.