JPS6191196A - Novel 5-fluoro-2'-deoxyuridine 5'-phosphate derivative and its salt - Google Patents

Abstract

NEW MATERIAL:A compound of formula I (R<1> is alkylammonio, cyclic ammonio; R<2> is oxide, hydroxyl; R3 is residue of 1-30C aliphatic carboxylic acid; A is lower alkylene) and its salt. EXAMPLE:5-Fluoro-2'-deoxy-3'-O-palmitoyl-beta-uridine-5'-(2-trimethyla mmonioethyl)- phosphate hydrate. USE:Antineoplastic: the decomposition of the compound in vivo is inhibited to develop high activity with low toxicity. PREPARATION:For example, the reaction of a compound of formula II such as 5-fluoro-2'-deoxy-3'-O-palmitoyl-beta-uridine-5'-(2-bromoethyl)phosphate with an alkylamine or cyclic amine, which is converted into R<1>, such as trimethylamine is carried out in an inert solvent such as toluene. The compound of formula II is novel and obtained, e.g., from a compound of formula III and another compound of formula IV.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a novel 5-fluoro-2'-deoxyclidine-51-phosphate derivative represented by the general formula and its derivatives.

The compounds of the present invention and their salts have strong antitumor activity and low toxicity, so they are useful as antitumor agents.

[Conventional technology]

Conventionally, 5-fluoro-2'-thioxy-β-uridine (
It is known that 5-fluorouracil (commonly known as FudR) has stronger cell-killing properties than 5-fluorouracil (commonly known as 5-Fu) in vitro [C, He1d@l
b@rger @tal, 'Cancer R
es, 282529-2558 (1961).

Furthermore, FudR is 5-fluoro-21-
Thioxyuridine-5'-monophosphate (commonly known as Fd
uMP), which inhibits thymidylate synthetase and is said to exert anticancer effects by inhibiting DNA synthesis [C,H*1del
b*rger at ml; Mol.

Pharmcol, , ,i, 14-30 (1965
)). but.

Clinically, FudR is only as effective as 5-Fu, and moreover, it is highly toxic (and is currently only used as a kinetic agent in the United States. (PHYSI)
CIANS' DESK REFERENCE52 e
dlton, 1587 (197B)).

[Problem that the invention seeks to solve]

In vitro, FudR is excreted rapidly and is not persistent, and is easily degraded by nucleoside phosphorylase and metabolized as α-fluoro-β-alanine via 5-Fu (C, H @ld
@lbsrger; Cancer Rem.

2, 1549-1569 (1970)"l, it has the disadvantage that it no longer exhibits its properties as a time-dependent antimetabolite that has a thymidylate synthase inhibitory effect.Also, although FduMP is an active form of FudR, By itself, it is not taken into the cell, but once it becomes FudB outside the cell, it enters the cell and turns into the active form FduMP again, exhibiting anti-ll [lls activity (R, N*Hunston@
ta1. ; J* Mad.

Che m-2'' 440-444 (1984)'), FduMP also had the same drawbacks as Ft+dR.

[Means for solving problems]

Under these circumstances, the present inventors have developed a drug that suppresses decomposition in vivo, has strong antitumor activity, and has low toxicity.
As a result of intense research to discover t+dR derivatives, Fud
H+ at the 5'-position of lH has the meaning indicated. ) The present inventors have completed the present invention by discovering that Fdt+MP derivatives represented by the general formula (]), which can be said to be a type of phospholipid, and salts thereof have the desired properties.

The compounds of the present invention will be explained in detail below.

As the alkylammonio group in R1.

Specific examples thereof include tri-C8-4 alkylammonio groups such as trimethylammonio, triethylammonio, dimethylethylammonio, diethylmethylammonio, triflobylammonio, and tributylammonio. In addition, as the R-shaped ammonio group,
Specifically, for example, pyridinio.

Viridasinio, pyrimidinio, 2I (-imidazorio,
Examples include 2H-pyrazolio, 1-methylbiperidinio, 4-methylmorpholinio, and the like. Furthermore, the cyclic ammonio group has carbon atoms such as methyl, ethyl, propyl, and butyl.
1-4 Alkyl group: di-C2-4 alkylamino group such as dimethylamino, diethylamino, ethylmethylamino, dipropylamino, dibutylamino, etc. Alkylamino group: may be substituted with a halogen atom such as fluoro, chloro, furomo, etc. V represents an oxide or hydroxyl group, and when R1 is an oxide group, the oxide group usually forms an inner salt with the alkyl ammonio or cyclic ammonio group in R1.

In addition, C1~., aliphatic carboxylic acid residues in n''FC include C, ~3 saturated or Cl-rich unsaturated aliphatic carboxylic acid residues.

As a C1-turtle saturated aliphatic carboxylic acid residue.

For example, formyl or acetyl, propionyl,
Butyryl, valeryl, hexanoyl, octanoyl, decanoyl, lauroyl, myristoyl, valmitoyl,
C surprises such as stearoyl, arachitoyl, pehenoyl,
go7/l/kayl group is mentioned, CB-@. Examples of unsaturated aliphatic carboxylic acid residues include C-carbons such as palmitoleoyl, oleoyl, lil'oyl, lil'noyl, etc.
,~1. Examples include alkenoyl groups.

Examples of lower alkylene groups in humans include linear or branched c
Examples include 1--alkylene groups.

In addition, as a salt of the compound represented by general formula (1),
Any drug may be used as long as it is pharmacologically acceptable; specifically,
For example, salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid; salts with organic acids such as 5-acetic acid, lactic acid, tartaric acid, methanesulfonic acid, benzenesulfonic acid; P) luenesulfonic acid; Examples include salts with alkali metals such as potassium; salts with alkaline earth metals such as magnesium, calcium, and barium.

Specific examples of the substituents R1, aS and A, and the salt of the compound represented by the general formula (1) have been given above.

The present invention is not limited to these.

Furthermore, the present invention includes optical isomers and geometric isomers of the compound represented by general formula (1) and its salts, and further includes all hydrates and crystal forms.

Next, the salt represented by the general formula (1) of the present invention can be used as a leaving group in the preparation )・Rogen atom 5benzenesulfonyloxy, p
-) arylsulfonyloxy groups such as luenesulfonyloxy.

Examples of the compound represented by the general formula (■) include tertiary alkylamines or cyclic amines that can be converted into alkylammonio or cyclic ammonio groups as explained in nl.

As a salt of a compound represented by general formula (It).

Examples include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as magnesium, calcium, and barium. Also, the general formula (
Examples of the salts of the compound represented by (maro) include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, and sulfuric acid; salts with organic acids such as methanesulfonic acid, benzenesulfonic acid, and p-)luenesulfonic acid. Examples include.

Next, this manufacturing method will be explained in more detail.

The reaction between the compound represented by the general formula (It) or its salt and the compound represented by the general formula (1) or its salt is usually carried out in a solvent inert to the reaction. Solvents used in this reaction include, for example, ketones such as acetone and methyl ethyl ketone; ethers such as diethyl ether, diisopropyl ether, tetrahydro7rane, and dioxane; aromatic hydrocarbons such as benzene and toluene; 1,1- Dichloroethane.

Examples include halogenated hydrocarbons such as methylene chloride and chloroform, and two or more of these may be used in combination. Further, when the compound represented by the general formula (1) is a liquid, it may be used in excess to serve as a solvent itself.

Further, the reaction temperature and reaction time are not particularly limited, but the reaction is usually carried out at room temperature to 80°C.

It can be completed in 5 minutes to 72 hours.

The compound represented by the general formula (1) or the salt thereof is used in an amount equal to or more than the amount of the compound represented by the general formula (1) or the salt thereof.

The compound represented by the general formula (1) may be obtained as a salt with the eliminated X, but after one reaction, if necessary, it may be treated with an ion exchange resin or silver ions to form the corresponding inner salt. It can also be converted into other salts by known methods. After such operations, general formula (1) can be obtained by appropriately combining ordinary purification methods such as silica gel column chromatography, ion exchange resin column chromatography, or recrystallization.
The compound represented by or its salt can be isolated and purified.

Furthermore, if stereoisomers exist, they can be further isolated according to a conventional optical resolution method, if necessary.

In addition, the reaction conditions in the above production method are such that the raw material, the compound represented by the general formula (1) or its salt, is a new compound, and for example, the method known per se as shown below or the general formula As the reactive derivatives of the compounds represented by (IV) and (Vl), specifically,
Mention may be made of reactive derivatives such as phosphorohalides, phosphorylimidazole, phosphoryltriazole forms, and the like.

Next, each manufacturing method will be explained in more detail.

Production method 1 The reaction of the compound represented by general formula (IV) or its reactive derivative with the compound represented by general formula m is usually carried out in the presence of a solvent inert to the reaction. Examples of the reagent used in this reaction include ketones such as acetone and methyl ethyl ketone; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran, and dioxane; nitriles such as acetonitrile and propionitrile; ethyl heptaacetate and acetic acid. Esters such as butyl; halogenated hydrocarbons such as 1,1-dichromitane, methylene chloride, and chloroform; aromatic hydrocarbons such as benzene and toluene; meformamide, N, N-
Examples include amides such as dimethylacetamide,
It is also possible to use a mixture of two or more of these solvents.

This reaction can also be carried out in the presence of a base. Examples of the base used here include triethylamine, tripropylamine, tributylamine, N-methylmorpholine, and N.

N-dimethylaniline, N, N-diethylaniline,
Examples include organic bases such as pyridine, 2,6-lutidine, and quinaldine. The amount of the base to be used is at least equimolar to the compound represented by the general formula (V), preferably from 1.0 to 1.0.
It is 2.0 times the mole.

Furthermore, when the compound represented by the general formula (IV) is used as a free acid, a suitable condensing agent can be used. As a condensing agent, N, N'-dicyclohexylcarbodiimide, N-cyclohexyl-Nl-morpholinoethylcarbodiimide, N-cyclohexyl-N'-(4-diethylaminoneclohexyl)carbodiimide, N-ethyl-h'- N,Nl-disubstituted carbodiimides such as (5-dimethylaminopropy/L/)carbodiimide.

°triphenylphosphine-2j ”-')
, ) Rudisulfide 0 Benzenesulfonyl chloride U )
'°2a6-) Liisobrobylbenzenesur? ! ! The amount of condensing agent used is as follows: Honyl chloride etc.
In addition, the reaction temperature and reaction time are not particularly limited,
The reaction is preferably carried out at -50 to 100 C1 under water cooling to room temperature, and in this case the reaction is usually completed in 10 minutes to 48 hours.

The compound represented by general formula (IV) or its reactive derivative is used in an amount equal to or more than the same molar amount, preferably 10 to 2.0 times the molar amount of the compound represented by general formula (y).

Production method 2 The reaction between the compound represented by the general formula (VJ) or its reactive derivative and the compound represented by the general formula (11) is as follows:
Usually in the presence or absence of a base.

The reaction is carried out in the presence of an inert solvent. The solvent, base, condensing agent, reaction conditions, etc. in this reaction are the same as in Production Method 1.

After carrying out the reaction of the above production method 1 or production method 2, the general formula (
The compound represented by (2) can be isolated and purified using operations such as column chromatography and/or recrystallization. Furthermore, if stereoisomers exist, they can be further isolated according to a conventional optical resolution method, if necessary. Among the compounds represented by the general formula (■), when R1 is a group other than a hydroxyl group,
Furthermore, the compound represented by general formula (1) or a salt thereof can be obtained by hydrolysis or removal of the hydroxyl protecting group by applying a known method. In this case, the compound represented by the general formula (■) may be isolated and used, or it may be directly converted into the compound represented by the general formula (1) or a salt thereof in the reaction system without isolation.

〔Effect of the invention〕

Next, the pharmacological effects of representative compounds of the present invention will be described.

1) Antitumor effect Group of 8 ddY mice (male, 5 withdrawn, 1 body weight approx. 255
+), Ehr;leh Carelnoma
5 x 10' small breasts were implanted subcutaneously in the inguinal area. A test compound dissolved or suspended in physiological saline was continuously administered intraperitoneally once a day for 6 days starting from the first day after transplantation. FudR was used as a control compound, and the control group received only physiological saline. The weight of the tumor was measured on the 12th day after transplantation, and the ratio ('r/
The results are shown in Table 1.

Table 1 2) Mouse Acute Toxicity Test A group of 5 ddY mice (male, 5 withdrawn) and a test compound dissolved or suspended in C1 physiological saline were each administered once intraperitoneally. On day 14 after administration, the mice were determined to be alive or dead, and the LDse value was calculated.

The results are shown in Table 2FC.

Table 2 As is clear from the above results, general formula (1) of the present invention
The compound represented by and its salts have excellent antitumor activity and low toxicity, so they are considered anti-Il! [It can be said that this compound is useful as a flk agent.]

When the compound represented by the general formula (1) of the present invention and its salts are used as a medicine, they can be prepared by themselves or mixed with appropriate additives such as pharmaceutically acceptable excipients, carriers, and diluents, and prepared into tablets, capsules, etc. agent.

It can be administered orally or parenterally in the form of granules, powders, injections, or suppositories. The dosage is.

The dose is usually about 1 to 1.50 ''f per day for adults, and is administered once or in several doses, and the dose is appropriately selected depending on age, body weight, and symptoms vc.

〔Example〕

Next, the present invention will be explained with reference to reference examples and examples, but the present invention is not limited thereto.

Reference Example 1 2.6-Lutidine α267 and 2-bromoethyl dichlorophosphate 0.589 were dissolved in anhydrous tetrahydrofuran IQm, and while stirring under water cooling, 5-
Fluo a-2'-deoxy-5'-〇-valmitoyl-β-uridine 0.97 F in anhydrous tetrahydrofuran 1
A solution containing 0 ml of VC is added dropwise over a period of 10 minutes.

After completion of the dropwise addition, the reaction was allowed to proceed overnight with stirring at room temperature, and then the reaction solution was concentrated under reduced pressure. Add 1Qa of water to the resulting residue.
l, chloroform 20117 and triethylamine 2
ml and stirred for 1 hour under water cooling and further stirred for 1 hour at room temperature. Then, the pH was adjusted to to with dilute hydrochloric acid, 20aj of methanol was added, and the organic layer was separated. The solvent is distilled off under reduced pressure, and the resulting residue is purified by column chromatography (Wako silica gel C-200, developing solvent: chloroform:methanol = 50:1 to 10:1'') to give a white amorphous product. 5-fluoro-27-dioxy-3'-0-valmitoyl-β-uridine-5'-(2
-bromoethyl)phosphate-) 1059 (yield 78%)
get.

In(KBr)3, 2920,2B50,17
20,1700°1680,1460,1355,12
50゜1180.1100,1060,102ONMR
102ON, :CD11OD=2:1 mixed solvent)
δ value; 0.89 (t, 5H, J=5H1), t0
3-1.80 (m, 26H).

2.00~+, 6o(m, 41.3.5s(t, 2H
, J=6Hz).

3.91-4.56 (m, 5H), 5.16-5
．． 50 (m, IH).

608-6.48 (m, IH), 7.81 (d,
IH, J = 6 Hz) Reference Example 2 26-lutidine 0.26 g, 2-bromoethyl dichlorophosphate 0.589 and 5-fluoro-21-deoxy-3'-〇-stearoyl β-uridine t03
If the reaction is performed in the same manner as in Reference Example 1 using g, and the front loading is performed,
White amorphous 5-fluoro-21-deoxy-3'-
1.059 of 〇-stearoyl-β-uridine-57-(2-bromoethyl)phosphate (yield 75%) is obtained.

1R(KBr)cs+, 2920.2B50.1
720.1700°1680, 1460.1555.1
250°1180, 1100.1050.102ONM
B (102ON2CD10D=2:1 mixed solvent) δ value;
0.89 (t, 3H, J=5H1), 1.05~
185 (rn, 30H).

2.05~22-60(,4)I), s, 5s(t
, zn, J=6Hz)75.91~4.57(rn
, 5H), 5.20-5.50 (m, IH).

6.10~&50 (m, IH), 7.86 (d, I
H, J=6Hz) Reference Example 3 2.6-lutidine α269.2-bromoethyl dichlorophosphate 0.5BS' and 5-fluoro-21-
Deoxy-61-0-oleoyl-β-uridine to2
If you react and purify in the same manner as in Reference Example 1 using
waxy 5-fluoro-21-deoxy-s'-o
105 g (yield 74%) of -oleoyl-β-uridine-5'-(2-bromoethyl)phosphate are obtained.

IR(KBr) m-'; 2920. 2850.
1720°1670, 1460. 1250°1180, 1100. 1060°02O NMR (CDC1 bird: CDa OD=2:1 mixed solvent) δ value; o, +39 (t, 5H, J=5) I engineering)
.

t O5mt 80 (mt 22 H)p 1.80
~2-60 (rr+, 8 H) 73.55 (t, 2
H, J=6Hz), 3.95-4.53(m, 5H
).

5.08-5.50 (m, 3H), 6.10-6.
44 (m, iH).

185 (d, I H, J = 6 Hz) Reference example 4 2.6-lutidine 0.26 F, 2-bromoethyl dichlorophosphate 0.589 Oyohi 5-fluoro 2I
-deoxy-3'-〇-hexanoyl-β-clidine 0
．． 849 and purified in the same manner as in Reference Example 1, an oily 5-fluoro-21-deoxy-3'-〇-hexanoyl-β-uridine-5'-(2-bromoethyl)phosphate)0. 90 PC yield (85%) is obtained.

-1°1B to) α, 2950. 2 B 60. 1
720°1660, 1460. 1260°1200, 1100. 1050°101 O NMR (CDCIs: CD=2:1 mixed bath medium) δ value: 0.89 (t, 5H, J=5Hz),
1.0 to 1.90 (m, 6H).

2.02-22-60 (, 4H), 5.53 (t,
2H, J=6HL).

&95~4.53 (m, 5H), 5.15~5.
55 (m, IH).

6.05-6.45 (rn, IH), 7.5o (d
, 1)], J=lZ) Example 1 Midoyl-β-uridine-5'-(2-)rimethy5-fluoro21-deoxy-3'-〇-valmitoyl-β
- Dissolve 1 g of uridine-5'-(2-bromoethyl)phosphate in 20 jlJ of toluene, add anhydrous trimethylamine 2-, and react in a bomb/roll at 30°C for 8 hours. Then, the reaction solution was concentrated under reduced pressure,
The obtained residue is dissolved in 50 aJ of a mixed solvent of chloroform and methanol (1:1) and washed twice with 201117 water. The solvent was distilled off under reduced pressure, and the resulting residue was subjected to column chromatography (Wako Silikage/l/C
-2Q O1 developing solvent; If purified with chloroform:methanol:water=(S5:25:4), a white amorphous 5
-Fluoro-2'-deoxy-3'-0-valmitoyl-β-uridine-5'-(2-)limethylammonioethyl)phosphate hydrate o, 9sy (yield 95%)
get.

Melting point: 230-265°C 1 R (KB r ) ar' ” 34507
2920/ 2850/172 et al. 1670, 1a6
Also 1250, 1090. 1060° 7 O NMR (CDCIs: CD@0D=2:1 mixed solvent) δ value; 0.88 (t, 3H, J=5H,
1.00-1.85 (m, 26H).

2.02-2.60 (rII,'H), 5-27
(m, 9H) 76.50-3.85 (rn, 2H),
5.85-4.60 (rn, 5H).

5.15~5.50 (mt IH)7 6.05~
lh 45 (my 1H) Js, o3 (d, IH, J
= 6 Hz) Example 2 105 g of 5-fluoro-2'-deoxy-3I-0-stearoyl-β-uridine-5'-(2-bromoethyl/I;) phosphate and anhydrous trimethylamine2. O
Using d, if reacted and purified in the same manner as in Example 1,
5-fluoro-2'-deoxy-3'-〇-stearoyl-β-uridine-5'-(2-)IJ methylammonioethyl) phosphate hydrate α999 (contains 95%
).

Melting point; 235-240°C (decomposition) 1R(KBr)cr'; 5430. 292Q,
2B50°1720, 1670. 1460°1255, 1090. 1060° 7 O NMR (CDCteasy: CD nose 0D=2:1 mixed solvent) δ
Value: 0.8B (t, 3H, J=5Hz) to 1.85 (m, 50H).

2.02-2.60 (m, 4H), 3.
27 (m, 9H).

5.50～585 (1”72H)p 5.85～
4.60 (''p 5 H)s5.15~5.50(
m, 1K), 6.05~&45 (U1H).

B-02 (dt IH, J = 6 H&) Example 3 Oil-β-uridine-5'-(2-trimethyl77monioethyl/L/)phosphate ohydrate 5-fluoro-
2'-deoxy-3'-〇-oleoyl-β-uridine-5'-(2-bromoethyl)phos7a) 0.98
9 and 2.0 m of anhydrous trimethylamine and purified in the same manner as in Example 1, 5-fluoro-7
, 1-deoxy-3'-〇-oleoyl-β-urisia 5'-(2-)limethylammonioethyl)phosphate hydrate 0.689 g (yield 70%) was obtained.

Melting point; 220 to 225°C (decomposition) 1111 (KBr) ex, 3430. 292
0. 2B50°1710, 1660. 146 Q
.

1240, 1170. 1085°1060, 965 NMR (CDC1-CDMOD=2:1 mixed bath medium) δ
Value; 0.89 (tp 31'g, J=5 Hz)p
102-1.85 (U22H).

t85~2.56(re, 8H), 3.28(s
i, 9H).

6.48~5L86 (my2H)t 5.86~
4.56 (m, 5H).

5.09~5-53 (my3H)76.01~6
-40 (my IH).

8-04 (at 14 J = 6 Hz) Example 4 5-Fluoro-2'-deoxy-3'-〇-hexanoyl β-uridine-5'-(2-bromoethyl)phosphate 0.85P and anhydrous trimethylamine 2.0 −
was reacted in the same manner as in Example 1, and then extracted with 20 ml of n-butanol.
and purified by silica gel column chromatography (Wako silica gel C-200, developing solvent: chloroform:methanol:water=65:25:4) to obtain white amorphous 5-fluoro-2'-deoxy-3I- 0.729 (yield: 85%) of 0-hexanoyl-β-uridine-5'-(2-)') methylammonioethyl) phosphate hydrate is obtained.

Melting point: 205-210°C (decomposition) 1 R (KB r ) cmp! +450.
2920. 2850°1720, 1660. 14
60°1240, 1165. 10 B 5゜1050,
96 O NMR (CDCIs-CDmOD=2:1 mixed solvent) δ
Value; 0.89 (t, 5H, J=5HS), 1.07
~188 (rn, 6H).

2.08~2.55(m, al(), x2'4(
a, 9)1).

148-175 (m, 2H), 3.86-4.
55 (mt 5 H) 75.13-5.42 (yn,
IH), 6.07~&43(m, 1)I).

7.98 (d. Add 0.21 dlIC bath dissolved 4-dimethylaminopyridine to 70
React at ℃ for 4 hours. Then, the solvent is distilled off under reduced pressure, and the resulting residue is dissolved in a mixed solvent of chloroform and methanol (20117) and washed successively with 5d of diluted hydrochloric acid and 5ILt of water. The solvent was distilled off under reduced pressure,
The resulting residue was purified by column chromatography (Wako silica gel C-200° developing solvent; chloroform:methanol:water = 65:25:1-2) to obtain white amorphous 5-fluoro-21-deoxy. -5'-0-valmitoyl-β-uridine-51-[2-(4-dimethylaminopyridinio)ethyl]phosphate hydrate 0.
129 (yield: 41%).

Melting point: 210-215°C (decomposition) 1 R (KB r ) cIT' p 3400J
2910x 2840/1710, 1640.
1560°1450, 1240. 1170°IQ +30. 1050. 82 DNMRCCD
CI Hatake) δ value: α88 (t, 3H, J=5Hz), toa~t87 (
m, 26H).

2.06-2.55 (m, 4H), 5.24 (m,
6H).

560-3.89 (m, 2H), 3.96-a, 5
1 (mt 7111 > p5.16-5.44 (m,
IH), 6.08-441 (m, IK).

Claims (1)

[Claims] General formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ In the formula, R^1 represents an alkylammonio or cyclic ammonio group; R^2 represents an oxide or hydroxyl group; R
^3 is C_1_~_3_0 aliphatic carboxylic acid residue;A
represents a lower alkylene group. 5-fluoro-2'-deoxyuridine-
5'-phosphate derivatives and salts thereof.