JP2884170B2 - Anticancer agent containing 5-deazaflavin compound as active ingredient - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION INDUSTRIAL APPLICATION The present invention relates to a compound of the general formula I (Wherein, R ₁ represents a hydrogen atom, an alkyl group, a halogen-substituted alkyl group, a carboxy-substituted alkyl group, an alkyloxycarbonylalkyl group, or a phenyl group; R ₂ represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group, An amino group, a phenyl-substituted alkylamino group, a hydroxy-substituted alkylamino group, an alkoxy group, a pyridyl group, a phenyl group, or a phenyl group substituted with one of a halogen atom, a nitro group, a lower alkoxy group, and a trifluoromethyl group the stands, R ₃ is a hydrogen atom, a halogen atom, a lower alkyl group, a hydroxyl group, a nitro group, a cyano group, a lower alkoxy group, a phenyl-substituted lower alkoxy group, a lower alkylamino group, a phenyl-substituted lower alkylamino group, or a lower It represents an alkylsulfonyl group, R ₄ Represents a hydrogen atom or a halogen atom, X is a sulfur atom, an oxygen atom, or represents a radical = N-R _5.

Here, R ₅ is an alkyl group, a cycloalkyl group, a phenyl-substituted lower alkyl group, a phenyl group, a phenyl group substituted with one of a halogen atom or a lower alkyl group or a lower alkoxy group, a lower alkyl disubstituted phenyl group. , Naphthyl group, lower alkyldisubstituted aminonaphthyl group, or the following group (In this case, the general formula I is And R ₁ , R ₃ and R ₄ represent the meaning as defined above, and n
Represents an integer from 1 to 20. ). The present invention relates to an anticancer drug comprising one or more 5-deazaflavin compounds represented by the formula (1).

[Prior Art] Numerous compounds having various structures have been proposed as compounds having an anticancer effect. That is, alkylating agents such as cyclophosphamide, antimetabolites such as fluorouracil, antibiotics such as mitomycin, hormones such as mepithiostan, plant components such as vincristine, various radioisotopes,
A wide variety of other compounds have been used as anticancer agents. However, although these compounds exhibit an excellent anticancer effect, they also have some drawbacks in that they damage the normal cells to some extent, so that they have strong toxicity and side effects. For this reason, the development of low-toxicity anticancer drugs is one of the most important points in this field.

Meanwhile, riboflavin known as vitamin B ₂ is FAD in vivo (flavin adenine dinucleotide) and FMN
(Flavin mononucleotide) and so on, and functions as a coenzyme for the flavin enzyme. The main roles of flavin coenzyme (Fl) are substrate dehydrogenation, electron transfer, activation of oxygen,
It performs photoreduction, chemiluminescence, etc., and has the most versatile functionality among coenzymes. In particular, Fl is inflamed,
In recent years, there has been much interest in having a function of activating oxygen involved in phenomena such as cancer and aging.

The relationship between Fl and cancer has been phenomenally known for a long time (RS Rivlin, “Riboflavin”, Plenum Press,
New York (1975), p.369). For example, it is known that when a riboflavin-deficient feed is given to a tumor-bearing mouse, the growth of cancer is suppressed as compared to when a normal feed is given.
Also, the diethyl analog of riboflavin was
It has been reported that the growth rate of er carcinoma is retarded and that the growth of ascites tumor in tissue culture is suppressed by the addition of riboflavin. In the latter case, it is said that there is no effect in a dark room and the presence of light is required, and it is inferred that riboflavin generates active oxygen by light and acts on cancer cells.

Although it is not directly related to cancer control, recently FMN has been using benzo [a]
Pyrene was found to function as a carcinogenesis stopper (Hagihiko Yagi, Pharmacia, 22 , 158 (1986) and
AW Wood, JM Sayer, HL Newmark, H. Yagi,
DP Michard, DM Jerina, AH Conney, Proc. N
atl. Acad. Sci., USA, 79 , 5122 (1982)). It converts benzo [a] pyrene diol epoxide, the ultimate active substance derived from benzo [a] pyrene, into FMN
Is thought to be due to trapping. Meanwhile, Ko
In a study of skin carcinogenesis in mice with 7,12-dimethylbenzo [a] anthracene, rnberg et al. reported that there was a marked reduction in the carcinogenesis rate of the riboflavin-treated group compared to the non-treated group. (A. Kornberg, WE Pri
cer Jr., J. Biol. Chem., 182 , 763 (1950)).

[Problems to be Solved by the Invention] However, the carcinogenesis-suppressing effect of the above-mentioned riboflavin is not always satisfactory, and its carcinostatic effect is not yet known at all.

Thus, the present inventors have conducted intensive studies on the synthesis and anti-cancer action of a compound having a flavin skeleton, and as a result, it has been found that 5-deazaflavin derivatives in which N at the 5-position of riboflavin is equivalently converted to CH are excellent anti-cancer drugs. It was discovered that there was a compound showing activity, and the present invention was completed based on this finding.

[Means for Solving the Problems] The compound of the present invention is a novel compound represented by the following formula (I) and has a strong anticancer effect.

In the formula (I), R ₁ represents a hydrogen atom, an alkyl group, a halogen-substituted alkyl group, a carboxy-substituted alkyl group, an alkyloxycarbonylalkyl group, or a phenyl group. Among them, a halogen-substituted alkyl group means that any one of the carbon atoms of the alkyl is a fluorine atom, a chlorine atom,
It means an alkyl group substituted with a bromine atom or an iodine atom, and examples thereof include a 3-bromo-n-propyl group, a 4-bromo-n-butyl group, and a 5-bromo-n-pentyl group. A carboxy-substituted alkyl group is
It means an alkyl group in which any one of the carbons of the alkyl is substituted with a carboxyl group, and examples include a carboxymethyl group. The alkyloxycarbonylalkyl group means a group of the formula “C _m H _{2m + 1} OCOC _n H _2n −” (m and n are natural numbers), for example, an ethyloxycarbonylmethyl group (C ₂ H ₅ OCOCH _2- ).

R ₂ represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkylamino group, a phenyl-substituted alkylamino group, a hydroxy-substituted alkylamino group, an alkoxy group, a pyridyl group, a phenyl group, or a halogen atom or a nitro group or a lower alkoxy group or a trialkyl group; Represents a phenyl group substituted by one of fluoromethyl groups. Among them, the alkylamino group means a group of the formula “—NHC _n H _{2n + 1} ” (n is a natural number), and examples include a methylamino group and an octylamino group. Phenyl-substituted alkyl amino group and a hydroxy-substituted alkyl amino group, each formula "-NH-C _n H _2n φ" and formula "-NH-C _n H _2n OH" form of groups (n is a natural number, phi is a phenyl group Represents).
Examples of the former include a phenylmethylamino group and a 2-phenylethylamino group, and examples of the latter include a 2-hydroxyethylamino group. A phenyl group substituted with one of a halogen atom, a nitro group or a trifluoromethyl group is a phenyl group in which one of the arbitrary carbon atoms on the benzene ring is substituted with one of these atoms and groups. Means a group. For example, 3-
Examples thereof include a fluorophenyl group, a 4-fluorophenyl group, a 4-nitrophenyl group, a 4-methoxyphenyl group, and a 4-trifluoromethylphenyl group.

R ₃ is a hydrogen atom, a halogen atom, a lower alkyl group, a hydroxyl group, a nitro group, a cyano group, a lower alkoxy group, a phenyl-substituted lower alkoxy group, a lower alkylamino group, a phenyl-substituted lower alkylamino group, or a lower alkylsulfonyl group Represents Among them, a lower alkyl group means an alkyl group having 1 to 6 carbon atoms, and a methyl group is preferable. The lower alkoxy group means an alkoxy group having 1 to 6 carbon atoms, and particularly preferably a methoxy group. The phenyl-substituted lower alkoxy group means a group of the formula “—OC _n H _2n φ” (n is an integer from 1 to 6, φ represents a phenyl group), and a benzyloxy group (—OCH ₂ φ) Is particularly preferred. The lower alkylamino group means a group of the formula “—NHC _n H _{2n + 1} ” (n is an integer from 1 to 6), and preferable examples include a methylamino group,
And an ethylamino group. A phenyl-substituted lower alkylamino group refers to a group of the formula “—NHC _n H _2n φ” (n is an integer from 1 to 6 and φ represents a phenyl group), and a benzylamino group (—NHCH ₂ φ) is particularly preferred. The lower alkylsulfonyl group means a formula (integer of n from 1 to 6) "-SO ₂ C _n H _{2n + 1"} form of groups, methylsulfonyl group (-SO ₂ CH ₃₎ is particularly preferred .

The position of R ₃ may be at any position 6-9 position on flavin ring. R ₄ represents a hydrogen atom or a halogen atom, and the position of R ₄ may be any of positions 6 to 9 on the flavin ring.

X is sulfur represents atom, an oxygen atom, or a group = N-R _5. Here, R ₅ is an alkyl group, a cycloalkyl group, a phenyl-substituted lower alkyl group, a phenyl group, a phenyl group substituted with one of a halogen atom or a lower alkyl group or a lower alkoxy group, a lower alkyl disubstituted phenyl group, Naphthyl group, lower alkyldisubstituted aminonaphthyl group, or the following group (In this case, the general formula I is In and, R _1, R ₃ and R ₄ are the same as R _1, R ₃ and R ₄ in each of the above substituents represents the meanings defined above, n represents an integer of from 1 to 20. ). Among them, a phenyl-substituted lower alkyl group, the formula "-C _n H _2n phi" shaped groups (integer from n is 1 to 6, phi represents a phenyl group)
Means, for example, a benzyl group. . A phenyl group substituted with one of a halogen atom, a lower alkyl group or a lower alkoxy group is a phenyl group in which one of any carbon atoms on a benzene ring is substituted with one of these atoms and groups. A lower group means an integer having 1 to 6 carbon atoms. Examples are 4-chlorophenyl, 3-methylphenyl, 4
-Methylphenyl group, 4-fluorophenyl group, 2-fluorophenyl group, 4-methoxyphenyl group and the like. The lower alkyldi-substituted phenyl group means a phenyl group in which any two carbon atoms on a benzene ring are substituted with an alkyl group having 1 to 6 carbon atoms. In this case, the two lower alkyl groups are the same, May also be different.
Examples include 2,4-dimethylphenyl and 3,4-dimethylphenyl groups. The lower alkyldi-substituted aminonaphthyl group means a naphthyl group substituted by an amino group substituted by two alkyl groups having 1 to 6 carbon atoms, for example, 8- (N, N-dibutylamino) -α- And a naphthyl group.

R ₅ based Where the compound of formula I is a flavin dimer as follows:

In this case, n represents an integer from 1 to 20.

Specific examples of the compound of the present invention represented by the general formula (I) are shown in Tables 1 to 4.

The compound represented by the general formula (I) and used in the present invention can be produced by any one of the following production methods A to H.

6-N-substituted-aminouracil starting material (2)
Is produced by condensation of 6-chlorouracil with a primary amine in a conventional manner. The 6-N- thus obtained
The substituted-aminouracil (2) and the appropriate o-halogenobenzaldehyde (3) are heated under reflux in dimethylformamide (DMF). An appropriate heating time is 3 to 5 hours. The reaction is concentrated under reduced pressure and the residue is recrystallized from a suitable solvent to give the corresponding 5-deazaflavin (6). The arylbis- (6-N-substituted-aminouracil-5-yl) methane (5) formed as an intermediate on the way may be purified and further heated in DMF.

(Production Method A ') 3-unsubstituted (R ₁ = H) 5-
Deazaflavin (6) and equimolar ClCH ₂ COOC
₂ H _5, BrC ₈ H ₁₇ or Br (CH ₂₎ compounds such as _n Br by heating in the presence of potassium carbonate in DMF, 3-position -CH ₂ COO
_{C 2 H 5, -C 8 H} 17, - (CH 2) n group is 5-Deazafurabin substituted derivatives such as Br can be obtained.

Heating the 6-N-substituted-aminouracil (2) with the appropriate arylbenzaldehyde (3 ') in acetic acid under reflux provides an arylbis- (6-N
-Substituted-aminouracil-5-yl) methane (5 ') is obtained. (5 ') is dissolved in excess diethyl azodicarboxylate (DAD) in the presence of sulfolane and heated to give the corresponding 5-deazaflavin (7).

5-Deazaflavin (8) having a leaving group is heated and reacted with benzyl alcohols (9) in the presence of potassium carbonate. The reaction mixture is concentrated under reduced pressure, the residue is treated with water, filtered and recrystallized to obtain the desired benzyloxy-substituted-5-deazaflavin (10).

Preparation C: Oxidation of 5-deazaflavin derivative (11) with m-chloroperbenzoic acid to give 4a, 5-epoxy-5-deazaflavin (12). (12) a Vilsmeier reagent, i.e., DMF-POCl _3: treated with (5 1), is then precipitated cooled to 5-chloro-5- Deazafurabin (14).

Production method D: In the same manner as in Production method C, 4a, 5-epoxy-5-deazaflavin (12) is obtained. (12) is added to pyridine to reflux and the reaction mixture is concentrated in vacuo. The residue was diluted with ether to give 1,5-dihydro-5-deazaflavin-5.
-Get on (13). Subsequently, (13) was chlorinated using POCl ₃ to give the desired 5-chloro-5-deazaflavin (1
4) get

The 6-anilinouracil derivative (15) and the aldehydes (16) are melted or heated under reflux in DMF for a long time to obtain the corresponding 5-deazaflavin (17).

3-N-substituted-6-chlorouracil (18) and α, ω-
The diaminoalkanes are heated to reflux in n-butanol. The crystals obtained on cooling are recrystallized from acetic acid to give bis (uracil-6-ylamino) alkane (19). (1
9) and the o-halogeno or o-methoxybenzaldehyde derivative are heated to reflux in DMF. After cooling, the precipitated crystals are recrystallized from DMF to obtain the corresponding bis (5-deazaflavin-10-yl) alkanes (20).

Refluxing the 3-N-substituted-6-chlorouracil (18) and thiophenol in ethanol in the presence of KOH,
Or refluxing in pyridine to give 6-arylthiouracil (21). Then, (21) is treated with DMF-POCl ₃ ,
The corresponding 6-arylthio-5-formyluracil (2
2) get Further heating (22) in polyphosphoric acid followed by dilution with water gives 10-thia-5-deazaflavins (23).

The 3-N-substituted-6-chlorouracil (18) is heated to reflux with appropriate phenols in the presence of K ₂ CO ₃ to give 6
Obtaining phenoxyuracils (24). (24) DMF-PO
Cl ₃ (5: 1) to obtain the heat to give 5-formyl-6-phenoxy uracils (25) and 5-dimethylaminomethylene-6-phenoxy-3-phenyluracils (26) together. The reaction mixture is heated in polyphosphoric acid and subsequently diluted in ice to give the 10-oxa-5-deazaflavins (2
7) get

Hereinafter, the present invention will be described in more detail with reference to Production Examples and Examples. Of course, the invention is not limited by these examples.

Production Example 1 10-n-butyl-5-deazaflavin (Compound No. 1) 6-Nn-butylaminouracil (2.5 mmol) and o-bromobenzaldehyde (3.0 mmol) in DMF (20 ml)
And heated under reflux at 140 ° C for 1.5 hours.
5-ylmethane was obtained. This intermediate (0.5g, 0.83mmol)
Was heated to reflux in DMF (20 ml) for 3 hours, then the reaction solution was concentrated under reduced pressure, and the residue was recrystallized from ethanol to obtain 10-butyl-5-deazaflavin as yellow crystals (yield). 79%).

Melting point: 302 ° C Elemental analysis value (%): C ₁₅ H ₁₅ N ₃ O ₂ CH N Calculated value 66.9 5.6 15.6 Actual value 66.7 5.6 15.5 Production Example 2 7-nitro-10-n-butyl-5-deazaflavin (compound No. 2) 6-N-butylaminouracil (2.5 mmol) and 2-
Using bromo-5-nitrobenzaldehyde (3.0 mmol) as a starting material, 7-nitro-10-
n-Butyl-5-deazaflavin was obtained as yellow crystals (80% yield).

Melting point: 290 ° C Elemental analysis value (%): C ₁₅ H ₁₄ N ₄ O ₄ CH N Calculated value 57.3 4.5 17.8 Observed value 57.2 4.2 17.6 Production Example 3 8-Chloro-10-n-butyl-5-deazaflavin (compound No. 3) 6-N-butylaminouracil and 2-bromo-4-
8-Chloro-10-n-butyl-5-deazaflavin was obtained in the same manner as in Production Example 1 except that chlorobenzaldehyde was used as a starting material and the ethanol for recrystallization was changed to DMF.

Melting point:> 300 ° C. Elemental analysis value (%): C ₁₅ H ₁₄ ClN ₃ O ₂ CH N Calculated value 59.3 4.7 13.8 Actual value 59.2 4.5 13.7 Production Example 4 3,10-dimethyl-5-deazaflavin (Compound No. 25) 3-methyl-6-methylaminouracil (0.01 mol)
And benzaldehyde (0.01 mol) were refluxed in acetic acid for 1 hour. The reaction mixture was concentrated under reduced pressure and the residue was recrystallized from acetic acid to give 5,5 '-(phenylmethylene) bis-
(3-Methyl-6-methylaminouracil) was obtained as a colorless powder. This intermediate (0.001 mol) was mixed with diethyl azodicarboxylate (DAD) (0.81 g, 0.005 mol) and the mixture was heated with stirring at 180 ° C. for 30 minutes. After cooling, the mixture was diluted with ethanol and left overnight at room temperature to obtain yellow crystals. Recrystallized from ethanol to give 3,10
-Dimethyl-5-deazaflavin was obtained as yellow crystals. (Yield 57%).

Melting point: 327 ° C Elemental analysis value (%): C ₁₃ H ₁₁ N ₃ O ₂ CH N Calculated value 64.7 4.6 17.4 Observed value 64.6 4.65 17.35 Production Example 5 3,10-dimethyl-8-fluoro-5-deazaflavin (compound No. 30) 3-methyl-6-methylaminouracil 1.55 g (0.01 m
ol) and 1.24 g of p-fluorobenzaldehyde (0.01mo
The mixture of l) was refluxed for 1 hour in 30 ml of acetic acid. The reaction mixture was concentrated under reduced pressure and the residue was recrystallized from acetic acid to give 5,
5 '-(p-fluorophenylmethylene) bis- (3-
1.85 g of methyl-6-methylaminouracil) was obtained as a colorless powder (89% yield).

5,5 '-(p-fluorophenylmethylene) obtained
Bis- (3-methyl-6-methylaminouracil) 0.42
g (0.001 mol) in diethyl azodicarboxylate (DA
D) mixed with 0.87 g (0.005 mol) and the mixture
Heat with stirring for 0 min. After cooling, the mixture was diluted with ethanol and left overnight at room temperature to obtain yellow crystals. Recrystallization from ethanol gave 0.15 g of 3,10-dimethyl-8-fluoro-5-deazaflavin (yield 59
%).

Melting point: 325 ° C, m / e: 259 (M ⁺ ), NMR: δ (CF ₃ CO ₂ H), 3,68 (3H, s, 3-Me), 4.53 (3H, s, 10-Me), 7.65~8.70 (3H, aromatic H), 9.78 (1H, s, 5-H) elemental analysis _{(%): C 13 H 10} FN 3 O 2 C H N calculated 60.23 3.89 16.21 Found 60.4 4.0 16.0 preparation 68-benzyloxy-3,10-dimethyl-5-
Deazaflavin (Compound No. 33) 3,10-dimethyl-8-fluoro-5-deazaflavin
0.26 g (0.001 mol) and potassium carbonate 0.28 g (0.002 mol)
Was heated in 3 ml of benzyl alcohol at 90 ° C. with stirring for 10 hours. The reaction mixture is concentrated under reduced pressure,
The residue was treated with water. Recrystallized from acetic acid 8
0.32 g of pale yellow needles of -benzyloxy-3.10-dimethyl-5-deazaflavin were obtained (96% yield).

Melting point: 298 ° C, m / e: 331 (M ⁺ ) NMR: δ (CF ₃ CO ₂ H), 3,65 (3H, s, 3-Me), 4.36 (3H, s, 10-Me), 5.52 (2H, s, benzylic H) , 7.30~8.45 (8H, aromatic H), 9.56 (1H, s, 5-H) elemental analysis _{(%): C 20 H 17} N 3 O 2 C H N calculated 72.49 5.17 12.68 Found 72.6 5.15 12.55 Production Example 7 3-Methyl-10-phenyl-5-deazaflavin (Compound No. 43) A mixture of 6-anilino-3-methyluracil (2.5 mmol) and o-bromobenzaldehyde was treated with n-butanol ( 30 ml) under reflux for 3 hours. Upon cooling the reaction mixture, o-bromophenylbis- (6-anilino-
Crude crystals of (3-methyluracil-5-yl) methane were obtained. This was recrystallized from n-butanol (yield 90
%). Melting point: 258-261 ° C (decomposition). This intermediate (0.5g,
0.83 mmol) was heated under reflux in DMF (20 ml) for 3 hours, cooled, recrystallized from DMF to give 3-methyl-10-phenyl-5.
-Deazaflavin was obtained (95% yield).

Melting point:> 330 ° C.

Elemental analysis _{(%): C 18 H 13} N 3 O 2 C H N Calculated 71.3 4.3 13.9 Found 71.0 4.3 13.9 Production Example 8 5-Chloro-3,10-dimethyl-5-Deazafurabin (Compound No. 97) a 4a, 5-Epoxy-3,10-dimethyl-5-deazaflavin 3,10-dimethyl-5-deazaflavin (3.05 mmol)
Contains m-chloroperoxybenzoic acid (3.77 mmol)
Add to CHCl ₃ (5 ml) solution and stir at solution temperature. After 150 minutes, the reaction solution was concentrated under reduced pressure, and the residue was recrystallized from a DMF-ether solution to give 4a, 5-epoxy-3,10-dimethyl-.
5-Deazaflavin was obtained (yield 92%, melting point 217 ° C).

b) 5-Chloro-3,10-dimethyl-5-Deazafurabin 4a, 5-epoxy-3,10-dimethyl-5- Deazafurabin (2 mmol) of _{DMF-POCl 3 (5: 1} ) in a solution (3 ml) of ,
Heat at 90 ° C. for 2 hours and then cool to give 5-chloro-3,
10-Dimethyl-5-deazaflavin was obtained (yield 74
%).

Melting point: 247 ° C. Elemental analysis value (%): C ₁₃ H ₁₀ ClN ₃ O ₂ CH N Calculated value 56.63 3.66 15.24 Actual value 56.70 3.61 15.02 Production Example 9 5-m-fluorophenyl-10-butyl-5
-Deazaflavin (Compound No. 112) 6-N-butylanilinouracil (1 mmol) and m-fluorobenzaldehyde (1.5 mmol) were heated to reflux in DMF (20 ml for 5 hours to give 5-m-fluorophenyl-10-
Butyl-5-deazaflavin was obtained (22% yield).

Melting point:> 300 ° C.

Elemental analysis _{(%): C 21 H 18} FN 3 O 2 C H N Calculated 69.40 4.99 11.56 Found 69.51 4.74 11.55 Production Example 10 bis - (8-chloro-5 Deazafurabin -
10-yl) octane (Compound No. 129) a) Bis- (uracil-6-ylamino) octane 6-chlorouracil (6.8 mmol) and 1,8-diaminooctane (3.4 mmol) in n-butanol (20 ml) For 8 hours, and cooled after the reaction was completed. The mother liquor was distilled off under reduced pressure, and the residue was recrystallized from glacial acetic acid to give bis- (uracil-
Colorless crystals of 6-ylamino) octane were obtained (yield 80
%, Melting point> 330 ° C).

b) bis- (8-chloro-5-deazaflavin-10-
Il) octane bis- (uracil-6-ylamino) octane (1.4 m
mol) and 2,4-dichlorobenzaldehyde (3.4 mmol)
Was heated to reflux in DMF (20 ml) for 8 hours, and cooled after completion of the reaction. The mother liquor was distilled off under reduced pressure, and the residue was recrystallized from DMF to obtain bis- (8-chloro-5-deazaflavin-10-yl) octane as a yellow powder (yield: 52%).

Melting point:> 330 ° C.

Elemental analysis _{(%): C 30 H 26} Cl 2 N 6 O 4 C H N Calculated 59.5 4.3 13.9 Found 59.7 4.2 13.5 Production Example 11 10-thia-5- Deazafurabin (Compound No.
139) a) 6-Phenylthiouracil 6-Chlorouracil (0.01 mol) and thiophenol (0.01 mol) were refluxed in pyridine (30 ml) for 1 hour. The pyridine was removed under reduced pressure and the residue was treated with water to give crystals, which were filtered and washed with ether. Recrystallization from ethanol gave 6-phenylthiouracil as colorless needles (yield 92%, mp 275 ° C).

b) 6-phenylthio-5-formyluracil 6-phenylthiouracil (0.01 mol) was added to DMF-POCl ₃
Added to the mixture and heated at 90 ° C. for 2 hours. Ice water was added to the reaction mixture, and the precipitate was removed. Recrystallization from ethanol gave pale brown needles (yield 96%, mp> 300).
° C).

c) 10-thia-5-deazaflavin 6-phenylthio-5-formyluracil (0.005mo
l) is dispersed in polyphosphoric acid (3 ml) and the mixture is brought to 120 ° C.
For 2 hours. After cooling, the reaction mixture was diluted with water and the resulting crystals were separated. Recrystallization from DMF yielded 10-thia-5-deazaflavin in reddish brown (88 yield).
%).

Melting point:> 300 ° C.

Elemental analysis _{(%): C 11 H 6} N 2 O 2 S C H N Calculated 57.37 2.63 12.17 Found 57.58 2.83 12.53 Production Example 12 10-oxa-5 Deazafurabin (Compound No. 150) a) 6- phenoxy uracil 6-chlorouracil (0.01mol) and phenol (0.03m
ol) and K ₂ CO ₃ (0.05 mol) were stirred and refluxed in DMF (20 ml) for 10 hours. The reaction mixture was concentrated under reduced pressure and the residue was treated with ether to remove excess phenol. The crystals thus obtained were dispersed in water and neutralized with acetic acid to separate crude crystals. Recrystallized from ethanol
Phenoxyuracil was obtained as colorless crystals (yield 53%,
270 ° C).

b) 5-Formyl-6-phenoxyuracil 6-phenoxyuracil (0.005 mol) was converted to DMF-POCl ₃ (5
ml: 1 ml) and heated at 90 ° C. for 2 hours. The reaction mixture was poured into ice water, the resulting precipitate was separated and dried over phosphorus pentoxide in a desiccator. Since the obtained compound became unstable when recrystallized, it was used for the next reaction without purification.

c) 10-oxa-5-deazaflavin 5-formyl-6-phenoxyuracil (0.005 mol) obtained as above was dispersed in polyphosphoric acid (7 ml),
The mixture was stirred at 120 ° C. for 2 hours. After cooling, the reaction mixture was diluted with ice water, and the resulting crystals were separated and recrystallized from a mixed solvent of acetic acid: acetic anhydride (10: 1) to give yellow platelets.
-Oxa-5-deazaflavin was obtained (86% yield).

Melting point: 325 ° C.

Elemental analysis (%): of _{C 11 H 6 N 2 O 3} C H N Calculated 61.68 2.82 13.08 Found 61.57 2.93 12.92 The present invention compounds listed in the Table 1 ~ Table 4, Production Example 1 to manufacture Other compounds not mentioned in Example 12 were synthesized in the same manner from the corresponding starting materials by the respective production methods. Table 5 shows the physical properties of the compound of the present invention thus obtained.

(Note 1) C-5: δ value means the δ value (in CF ₃ CO ₂ H) of the proton bonded to the 5-position carbon of the 5-deazaflavin ring in NMR analysis.

(Note 2) Recrystallization was not performed due to instability.

(Note 3) The corresponding 5-hydroxy-5-deazaflavin is identified as that obtained by chlorination with POCl ₃ . (Elemental analysis was not performed.) (Note 4) The NMR data of the compounds of Compound Nos. 139 to 149 are as follows.

(Note 5) The UV and visible light absorptions of the compounds of Compound Nos. 150 to 153, 155, 156, 159 to 161 and the δ value of the proton bonded to the 5-position carbon of the 5-deazaflavin ring are as follows.

(Note 6) Compound Nos. 150 to 153, 155, 156, 159 to 161 are all mixed solvents of acetic acid and acetic anhydride (mixing ratio: 10:
Recrystallized from 1).

Since the compound of the present invention has antitumor activity against various tumor cells, it can be used as an anticancer agent.

The following pharmacological experiments explain that the compounds of the present invention have excellent antitumor properties. Pharmacological Experiments: Effect various cultured tumor cells (10 ⁴⁾ on the in vitro expansion of various cultured tumor cells and the culture medium containing the test compound so as to 200μl to each well of a 96-well plate (FALCON 3072) In addition, the cells were cultured at 37 ° C. in a 5% CO ₂ -95% air atmosphere for 72 hours. After the culture, 25 μl of an MTT [3- (4,5-dimethyl-thiazol-2-yl) -2,5-diphenyltetrazolium bromide] solution (2 mg / ml) was added to each well, and the mixture was further incubated at 37 ° C. for 4 hours. 5% CO ₂ -95% and incubated under air. After removing the culture solution, 150 μl of dimethyl sulfoxide was added to each well to dissolve the formed MTT-formazan, and the absorbance at 540 nm was measured using a microplate photometer to obtain an index of the number of cells. The inhibition rate was calculated by the following formula, and the concentration (IC ₅₀ ) of the test compound that inhibited by 50% was determined.

The various tumor cell lines were cultured in an RpMl1640 medium (Nissui Pharmaceutical) containing 10% FCS.

Table 6 summarizes the obtained IC ₅₀ values (μg / ml).

As is clear from Table 6, the compound of the present invention shows an excellent growth inhibitory effect on various cultured tumor cells and is useful as an anticancer agent.

The administration method of the anticancer agent of the present invention is not particularly limited, and a general method for tumor treatment can be applied according to various preparation forms, age of the patient, gender and other conditions, degree of disease, and the like. That is, glucose alone or alone as an injection
It is administered intravenously as a mixture with a normal replacement fluid such as amino acids, or is administered alone subcutaneously, intramuscularly or intraperitoneally. It can also be orally administered in the form of tablets, pills, solutions, suspensions, emulsions, granules or capsules.
Further, it can be administered into the rectum as a suppository and applied / instilled as an external preparation.

The dose of the anticancer agent of the present invention may be an amount capable of effectively inhibiting the target tumor, and may vary depending on, for example, the compound to be administered, the symptom of the tumor, the administration route, the dosage form and the like. Generally, a single dose of a compound of the invention will be from 0.1 to 500 mg / kg body weight, with an upper limit preferably of about 250 mg / kg body weight, more preferably of about 10 mg / kg body weight.
The dose is preferably 0 mg / kg body weight. In the case of an injection, the upper limit is often about 50 mg / kg body weight. 1 dose per day
The number is appropriately selected in the range of 1 to 6 times.

When the compound of the present invention is administered as an anticancer agent, it is used in the form of a general pharmaceutical preparation as described above. The preparation is prepared by using a diluent or excipient such as a filler, a bulking agent, a binder, a humectant, a disintegrant, a surfactant and a lubricant which are usually used. As the pharmaceutical preparation, various forms can be selected according to the purpose of treatment, and typical examples thereof include tablets, pills, powders, solutions, injections (solutions, suspensions, etc.). In molding into tablets, carriers widely known in the art can be used. For example, lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, crystalline cellulose, excipients such as silicic acid, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, Binders such as methylcellulose, potassium phosphate, polyvinylpyrrolidone, dried starch, sodium alginate, carten powder, laminaran powder, sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, monoglyceride stearate, starch, Disintegrating agents such as lactose, sucrose, stearin, cocoa butter, disintegrating inhibitors such as hydrogenated oil, quaternary ammonium salts, absorption promoting agents such as sodium lauryl sulfate, glycerin,
Examples include humectants such as starch, adsorbents such as starch, lactose, kaolin, bentonite, and colloidal silicic acid, and lubricants such as purified talc, stearates, boric acid powder, and polyethylene glycol. Further, the tablet can be a tablet coated with a usual coating, if necessary, for example, a sugar-coated tablet, a gelatin-encapsulated tablet, an enteric-coated tablet, a film-coated tablet or a multilayer tablet. In the case of molding in the form of pills, those conventionally known in the art can be widely used as carriers. For example, excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc, gum arabic powder, tragacanth powder, binders such as gelatin,
Disintegrators such as laminaran powder and agar powder are exemplified. For molding into a suppository form, conventionally known carriers can be widely used. For example, polyethylene glycol, cocoa butter, higher alcohol, esters of higher alcohol, gelatin and the like can be mentioned. When prepared as an injection, the solutions, emulsions and suspensions are preferably sterile and isotonic with blood. These liquids,
In preparing emulsions and suspensions, any diluent commonly used in this field can be used. For example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like can be mentioned. In this case, a sufficient amount of salt, glucose or glycerin to prepare an isotonic solution may be contained in the anticancer agent, and also a normal solubilizing agent, a buffer,
It may contain a soothing agent and the like. If necessary, coloring agents, preservatives, flavors, flavors, sweeteners and other pharmaceuticals may be added to the anticancer agent.

Examples of the anticancer agent using the compound of the present invention will be described below.

Example 1 (Tablets) (1) Compound No. 75 compound 5 g (2) Lactose 50 g (3) Corn starch 30 g (4) Microcrystalline cellulose 25 g (5) Methyl cellulose 2 g (6) Magnesium stearate 1 g Compound No. 75 compound, lactose , Corn starch and microcrystalline cellulose are thoroughly mixed, and a 5% aqueous solution of methylcellulose is added to granulate and dried. The magnesium stearate is mixed with the dried granules, and the mixture is compressed into a tablet using a tablet machine.
A tablet containing 5 mg of the active ingredient (1) per tablet is produced.

Example 2 (Capsule) (1) Compound No. 158, 5 g (2) Lactose, 120 g (3) Starch, 30 g (4) Talc, 5 g (5) Magnesium stearate, 2 g Finely powder all components to form a uniform mixture After sufficient stirring, the mixture is filled into hard gelatin capsules to produce capsules containing 5 mg of the active ingredient per capsule.

Example 3 (injection) (1) Compound No. 87 1 g (2) Propylene glycol 3 g (3) Polyoxyethylene sorbitan monooleate 2 g (4) Sodium metabisulfite 0.1 g (5) Methyl-paraben 0.18 g (6) Propyl-paraben 0.02 g (7) Water for injection 100 ml in total The above propylene glycol, polyethylene glycol, polyoxyethylene sorbitan monooleate, methyl-paraben and propyl-paraben are compound No. 87
Are mixed and stirred.

Sodium metabisulfite is added to the obtained solution, the total volume is further made up to 100 ml with water for injection, sterilized with a Millipore filter, and then filled into a glass ampoule to prepare an injection.

Continuation of the front page (56) References US Pat. No. 4,272,535 (US, A) German published document 3232621 (DE, A1) Heterocycl. Chem. , 24 (1), 251-2 (1987). Chem. Soc. , Perkin Trans. 1 (3), 561-5 (1984) Heterocycl. Chem. , 18 (7), 1329-34 (1981) (58) Fields investigated (Int. Cl. ⁶ , DB name) A61K 31/505 C07D 471/04 C07D 491/052 C07D 495/04

Claims (1)

(57) [Claims] 1. A 5-deazaflavin compound represented by the general formula I: (Wherein, R ₁ represents a hydrogen atom, an alkyl group, a halogen-substituted alkyl group, a carboxy-substituted alkyl group, an alkyloxycarbonylalkyl group, or a phenyl group; R ₂ represents a hydrogen atom, a halogen atom, a hydroxyl group, an alkyl group, An amino group, a phenyl-substituted alkylamino group, a hydroxy-substituted alkylamino group, an alkoxy group, a pyridyl group, a phenyl group, or a phenyl group substituted with one of a halogen atom, a nitro group, a lower alkoxy group, and a trifluoromethyl group the stands, R ₃ is a hydrogen atom, a halogen atom, a lower alkyl group, a hydroxyl group, a nitro group, a cyano group, a lower alkoxy group,
A phenyl-substituted lower alkoxy group, a lower alkylamino group, a phenyl-substituted lower alkylamino group, or a lower alkylsulfonyl group, R ₄ represents a hydrogen atom or a halogen atom, and X represents a sulfur atom, an oxygen atom, or a group = represent N-R _5. Here, R ₅ is an alkyl group, a cycloalkyl group,
A phenyl-substituted lower alkyl group, a phenyl group, a phenyl group substituted with one of a halogen atom or a lower alkyl group or a lower alkoxy group, a lower alkyl disubstituted phenyl group, a naphthyl group, a lower alkyl disubstituted aminonaphthyl group, or Base (In this case, the general formula I is And R ₁ , R ₃ and R ₄ represent the meaning as defined above, and n
Represents an integer from 1 to 20. ). An anticancer agent comprising one or more of the above as an active ingredient.