JPH0714955B2 - Novel camptothecin derivative - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel camptothecin derivatives. More specifically, the present invention has the general formula (In the formula, R ¹ represents a hydrogen atom or a C _1-4 alkyl group, R ² is an aldose residue, or —COOH group, —NH ₂ group or —OH group present in the group is Aldose protected by a lower aliphatic acyl group having 1 to 2 carbon atoms in the alkyl group, a benzoyl group, a trifluoroacetyl group, a lower alkyl ester group having 1 to 2 carbon atoms in the alkyl group, or a combination thereof. A residue, in which the bond between R ² and camptothecin represents the glycosidic bond between the oxygen atom on the A ring of camptothecin and the sugar-1 carbon atom) or an alkali metal in the sugar moiety thereof. It provides a salt, a quaternary ammonium salt, or an acid addition salt (such as a mineral acid salt or an organic acid salt).

Camptothecin is Camptotheca acuminata
Alkaloids extracted and isolated from Nyssaceae, etc.,
It is an antitumor substance that has a strong inhibitory action on nucleic acid synthesis, is characterized by rapid and reversible actions, and does not show cross resistance with existing anticancer agents. It is recognized that it has a strong anti-tumor effect on experimental transplant cancer such as L1210 and rat walker 256 sarcoma, but its usefulness as a drug is naturally limited due to its toxic effect is there.

The present inventors have made many attempts to obtain new camptothecin derivatives having improved pharmacological activity and toxicity by chemically modifying natural camptothecin. Among the camptothecin derivatives obtained so far, especially for the compound group having a hydroxyl group at the 10-position, there are interesting results that toxicity is reduced while maintaining superior antitumor activity. Is difficult to administer as a medicine because it is poorly soluble in water.

Conventionally, as one of the methods of making a camptothecin derivative water-soluble, a method of forming a sodium salt of a carboxylic acid by ring-opening of an E ring (lactone ring) is known. It has the drawback of being weakened to a fraction.

The present inventors have succeeded in obtaining a glycoside derivative of a hydroxyl group on the A ring (9-, 10-, 11-, and 12-positions) without cleaving the E ring (lactone ring).

The novel glycoside derivative according to the present invention can be produced as follows.

A camptothecin derivative having a hydroxyl group on the A ring is dissolved or suspended in a solvent such as acetone, dioxane, dimethylsulfoxide, dimethylformamide, dimethylacetamide, or tetrahydrofuran, preferably acetone, to which 4.0 to 5.0 equivalents of anhydrous potassium carbonate and an alkyl group are added. A lower aliphatic acyl group having 1 to 2 carbon atoms, a benzoyl group, a trifluoroacetyl group, a lower alkyl ester group having 1 to 2 carbon atoms in the alkyl group, or a halogenoaldose protected by a combination thereof ( As the halogen atom, chlorine or bromine atom, preferably bromine atom) is added in an amount of 4.0 to 8.0 equivalents, and the mixture is stirred vigorously at room temperature to the boiling temperature of the solvent for 3 to 24 hours, and then the reaction mixture is reacted. The insoluble material is removed by filtration, and the product is purified by silica gel column chromatography. Separated by chromatography and purified. Thus, a camptothecin glycoside compound having a protecting group on the glycoside portion is obtained.

Alternatively, as another method, a camptothecin derivative having a hydroxyl group on the A ring is dissolved in 2.0 to 2.5 equivalents of an aqueous sodium hydroxide solution or an aqueous potassium hydroxide solution, preferably an aqueous potassium hydroxide solution, and 2 equivalents of the above-mentioned Add an acetone solution of halogenoaldose having a protecting group, and
There is a method of reacting at 60 ° C for a certain time.

When the halogen atom at the halogenoaldose position and the acetoxy group at the 2-position are in the trans position, an orthoester derivative is produced.

Protected by a lower aliphatic acyl group having 1 to 2 carbon atoms in the alkyl group, a benzoyl group, a trifluoroacetyl group, a lower alkyl ester group having 1 to 2 carbon atoms in the alkyl group, or a combination thereof. Halogenoaldose (chlorine or bromine atom as halogen atom,
(Preferably a bromine atom) can be produced by a known method generally used in sugar chemistry.

That is, the number of carbon atoms in the aldose and the alkyl group is 1 to
2 lower acid anhydride and acidic catalyst (for example, zinc chloride, sulfuric acid, hydrogen chloride, perchloric acid, etc.) or basic catalyst (for example,
The reaction is conducted in the presence of sodium acetate, pyridine, etc.) to give a peracyl aldose (hereinafter, acyl means a lower acyl group having 1 to 2 carbon atoms in the alkyl group), and then hydrogen halide (hereinafter hydrogen chloride or bromide). Represents hydrogen)
Peracylhalogenoaldose can be produced by reacting the acetic acid solution of.

For example, 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide is available from Methods Carbohyd. Chem.,
According to the method described in 2,221 (1963), 2,3,4-tri-O-acetyl-α-L-fucopyranosyl bromide is
According to the method described in Carbohydrate Research 64,297 (1978), 2,3,6,2 ', 3', 4 ', 6'-penta-
O-acetyl-α-D-maltosyl bromide is J. Am. C.
hem.Soc., 51, 1820 (1929).

Further, as another method, a reaction between peracyl aldose and titanium tetrachloride, liquid hydrogen chloride, aluminum chloride, aluminum chloride-phosphorus pentachloride, zinc chloride-thionyl chloride, hydrogen halide-acyl halide, or aldose and acyl halide It can be produced by a reaction with.

Also, aldose was reacted with a lower alcohol having 1 to 2 carbon atoms in an anhydrous alkyl group containing hydrogen chloride to give an alkyl aldose (Fischer method), and then benzoyl halide was reacted with pyridine as a solvent under ice-cooling to give an alkyl peroxide. As benzoyl aldose, further hydrogen halide-
Perbenzoylhalogenoaldose can be prepared by reaction with acetic acid solution.

Alternatively, it can also be produced by reacting 1-acyl perbenzoyl aldose with hydrogen under anhydrous halogen in a solvent such as chloroform, ether or acetic acid.

In the case of aldose (uronic acid) having a carboxyl group in the molecule, the lactone derivative thereof is boiled under reflux in a lower alcohol having 1 to 2 carbon atoms in the anhydrous alkyl group or sodium methoxy in the corresponding alcohol. Sodium ethoxide, sodium hydroxide,
The carboxyl group is protected by a lower alkyl group having 1 to 2 carbon atoms in the alkyl group by a reaction with triethylamine or triethylammonium hydroxide, and then the above is used as a lower acid anhydride having 1 to 2 carbon atoms in the alkyl group. The alkyl peracyl halogenoaldose uronate can be prepared by acylating the hydroxyl group in the presence of the acidic catalyst or the basic catalyst, followed by reaction with a hydrogen halide-acetic acid solution.

For example, methyl (2,3,4-tri-O-acetyl-α-D
-Glucopyranosyl bromide) uronate is a method
It can be produced according to the method described in Carbohyd. Chem., 2, 335 (1963).

Further, in the case of an aldose having an amino group in the molecule (aminoaldose), an aminoaldose is heated with an acyl halide to form an O-peracylaminohalogenoaldose-hydrogen halide salt, and then trifluoroacetic anhydride or alkyl anhydride. A trifluoroacetamide or a lower acylamide peracyl halogenoaldose can be produced by reacting a lower acid anhydride having a group having 1 to 2 carbon atoms.

As another method, peracylaminoaldose / hydrogen halide is trifluoroacetic anhydride using a pyridine catalyst in a solvent such as methylene chloride, a lower acid anhydride having 1 to 2 carbon atoms in the alkyl group, or a halogenated salt. It can also be prepared by reacting with an acyl and then with a hydrogen halide-acetic acid solution.

For example, 3,4,6-tri-O-acetyl-2-deoxy-
2-Trifluoroacetamide-α-D-glucopyranosyl bromide is described in J. Org. Chem., 31, 507 (1966), J. Org.
It can be produced according to the method described in Chem., 32, 1821 (1967).

The protecting group for the glycoside compound of camptothecin, that is, the lower acyl protecting group, the benzoyl protecting group, the trifluoroacetyl protecting group, the lower alkyl ester protecting group is the same as the above-mentioned glycoside compound in sodium, methoxy, or acetone. De-methanol solution or
It is removed by subjecting it to mild alkaline hydrolysis with 0.1N aqueous sodium hydroxide. This hydrolysis product is acidified with an ion exchange resin (sulfonic acid-H ⁺ type) or 0.1N hydrochloric acid, and the product is recrystallized from a solvent such as methanol, ethanol, or ethanol-n-hexane, or
Alternatively, a suitable solvent (for example, separated by ion exchange resin or reverse phase chromatography, for example, ODS column etc. after purification, for example,
A glycoside compound of a camptothecin derivative can be obtained by recrystallization from methanol, ethanol, ethanol-n-hexane, etc.).

Furthermore, when the sugar moiety has an amino group, as an acid addition salt (salt such as mineral acid or organic acid) at the amino group,
When it has a carboxyl group, it can be obtained as its alkali metal salt or quaternary ammonium salt.

Hydroxycamptothecin as a raw material and its derivatives have a hydroxyl group capable of glycosidation at the 20th position in addition to the hydroxyl group on the A ring. However, in the glycosidation method as described above, the hydroxyl group at the 20th position is Does not undergo glycosidation. Therefore, no protective group is required for the 20th hydroxyl group.

Other glycosidation methods include, for example, a method of reacting hydroxycamptothecin and its derivative with the above-mentioned protected halogenoaldose in the presence of silver carbonate or silver oxide (Knigs-Knorr reaction), and aldose acetate being chlorided. Examples thereof include a method of reacting with heating using zinc or p-toluenesulfonic acid as a catalyst, a method of reacting the above-mentioned protected halogenoaldose in an alkaline solvent (pyridine or quinoline etc.), and the like.

In addition, as a method for removing the protective group, the
A method using 0.5N barium methoxide, a method using ammonia-saturated methanol and the like can be mentioned.

Of the camptothecin and the camptothecin derivative's A-ring hydroxyl group substitution products, which are raw materials for the production of the novel camptothecin derivative according to the present invention, the 9-position substitution product and the 12-position substitution product are camptothecin, or 9-nitro form or 12-nitro form of camptothecin. Can be produced through reduction and diazotization (see JP-A-59-51289).

10-hydroxycamptothecin or 7-lower alkyl-
10-hydroxycamptothecin is obtained by reacting camptothecin or 7-lower alkylcamptothecin with hydrogen peroxide in acetic acid to convert them into N-oxides.
The N-oxide is dissolved in a solvent such as dioxane, acetonitrile, chloroform, methylene chloride, glyme, diglyme or an arbitrary mixed solvent thereof, and ultraviolet rays are obtained by using a mineral acid such as sulfuric acid or perchloric acid or an organic sulfonic acid. It can be produced by treating it under irradiation (Japanese Patent Laid-Open No. 58-58).
-39683, JP-A-58-39684, JP-A-58-39685).

The 11-hydroxy-substituted compound can be produced through oxidation (aromatization), reduction, and diazotization using the 11-nitro compound obtained by nitration of the B-ring reduced tetrahydro compound of camptothecin as a starting material (JP-A-59). -51287 gazette). 11-Hydroxycamptothecin can be obtained by introducing a lower alkyl group at the 7-position thereof by the method developed by the present inventors. (JP-A-56-1587
(See Publication No. 86).

The present invention will be described in detail below with reference to examples.

Example 1 7-Ethylcamptothecin-10-yl 2 ', 3', 4 ', 6'
-Tetra-O-acetyl-β-D-glucopyranoside Method A: 7-ethyl-10-hydroxycamptothecin 1.57
g, 2,3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide 12.48 g and anhydrous potassium carbonate 3.01 g
Suspended in 300 ml of acetone and shake vigorously for 20
Boil to reflux for an hour. After completion of the reaction, the solvent is distilled off under reduced pressure, chloroform and water are added to the residue, and the mixture is shaken. The material insoluble in both layers is filtered off using Celite. The chloroform layer is separated, washed with saturated brine, dried over magnesium sulfate, and concentrated under reduced pressure.

The concentrate was subjected to medium pressure silica gel column chromatography (eluted with chloroform followed by chloroform / methanol (100/1)) and then recrystallized from chloroform-n-hexane to give 0.71 g of the title compound as colorless needles. Obtained as crystals.

Method B: 7-ethyl-10-hydroxycamptothecin 100m
After dissolving g in 0.1N potassium hydroxide aqueous solution 6.4 ml, 2,
A solution of 3,4,6-tetra-O-acetyl-α-D-glucopyranosyl bromide (210 mg) in acetone (5 ml) is added, and the mixture is reacted at room temperature for 2 hours. After filtering off the yellow substance that precipitates during the reaction, acetone is distilled off under reduced pressure at a low temperature. The resulting aqueous solution is shaken with chloroform and 0.1N hydrochloric acid. The chloroform layer is separated, washed with water, dried over magnesium sulfate, and concentrated under reduced pressure.

The concentrate was subjected to medium pressure silica gel column chromatography to obtain 50 mg of the title compound (eluting with chloroform followed by chloroform / methanol (100/1)).

MP191-192 ℃ (decomposition) SIMS: MH ⁺ 723 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.04 (t) 20-CH ₂ CH ₃ 1.40 (t) 7-CH ₂ CH ₃ 1.81 to 1.98 (m) 20-CH ₂ CH ₃ 2.06,2.07,2.08 , 2.11 (s × 4) 2 ′, 3 ′, 4 ′, 6′-OCOCH ₃ 3.07 to 3.24 (m) 7-CH ₂ CH ₃ 3.83 (s) 20-OH 3.98 (d × d × d) 5 '-H 4.22 (dxd) 6'-C H (H)-4.33 (dxd) 6'-CH ( H ) -5.20 to 5.42 (m) 1'-H, 2'-H, 3' -H, 4'-H 5.25 (s ) 5-CH 2 - 5.31 (d) 17-C H (H) - 5.75 (d) 17-CH (H) - 7.51 (d × d) 11-H 7.60 ( d) 9-H 7.64 (s) 14-H 8.20 (d) 12-H Example 2 7-Ethylcamptothecin-10-yl β-D-glucopyranoside 7-Ethylcamptothecin-10-yl 2 ', 3', 4 ', 6'
-Tetra-O-acetyl-β-D-glucopyranoside 60
0 mg was suspended in 60 ml of anhydrous methanol, 0.5 ml of 28% sodium methoxide-methanol solution was added thereto, and the suspension was stirred at room temperature for 3 days.
Let react for 0 minutes. After the reaction, 2 ml of water is added, and an ion exchange resin (sulfonic acid-H ⁺ type) is added thereto to make the liquid weakly acidic. After removing the resin by filtration and washing with methanol, the washings are combined and dried under reduced pressure. The residue was recrystallized from methanol to give the title compound (410 mg) as pale-yellow needles.

MP232-233 ℃ (decomposition) SIMS: MH ⁺ 555 ₁ H-NMR δ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ C H ₃ 1.31 (t) 7-CH ₂ C H ₃ 1.78 to 1.96 (m) 20-CH ₂ CH ₃ 3.12 to 3.25 ( M ) 7-C H ₂ CH ₃ , Glc.-H 3.32 to 3.39 (m) Glc.-H × 2 3.44 to 3.54 (m) Glc.-H × 2 3.70 to 3.80 (m) Glc.-H 4.68 (d × d) 6'-OH 5.10 (d) Glc.-OH 5.14 (d) 1'-H (J 1 ', 2' = 8.06Hz) 5.16 (d) Glc.-OH 5.32 (s) 5- _{CH 2 - 5.43 (s) 17} -CH 2 -, Glc.-OH 6.50 (s) 20-OH 7.29 (s) 14-H 7.58 (d × d) 11-H 7.74 (d) 9-H 8.11 (d ) 12-H Example 3 7-Ethylcamptothecin-10-yl 2 ′, 3 ′, 4′-tri-O-acetyl-β-L-fucopyranoside 7-ethyl-10 hydroxycamptothecin 1.57 g, 2,
3,4-Tri-O-acetyl-α-L-fucopyranosyl bromide (5.86 g) and anhydrous potassium carbonate (2.35 g) were added to acetone (30).
Suspend in 0 ml and boil under reflux for 4 hours with vigorous stirring. After completion of the reaction, the solvent is distilled off under reduced pressure, the residue is dissolved in chloroform, and the insoluble substance is removed by filtration using Celite. The filtered material is washed with chloroform, the filtrates are combined and concentrated under reduced pressure. The concentrate was subjected to medium pressure silica gel column chromatography (eluted with chloroform and then with chloroform / methanol (100/1)), and then recrystallized from ethanol to obtain 1.72 g of the title compound as colorless needle crystals. .

MP258-260 ℃ (decomposition) SIMS: MH ⁺ 665 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.04 (t) 20-CH ₂ C H ₃ 1.34 (d) 6′-CH ₃ 1.40 (t) 7-CH ₂ C H ₃ 1.81 to 1.98 (m) 20-C H ₂ CH ₃ 2.05,2.11,2.24 (s × 3) 2 ′, 3 ′, 4′-OCOCH ₃ 3.06 to 3.21 (m) 7-C H ₂ CH ₃ 3.86 (s) 20-OH 4.09 (q) 5 '-H 5.20 (d × d) 3'-H 5.25 (s) 5-CH 2 - 5.27 (d) 1'-H (J 1', 2 '= 8.06Hz) 5.31 (d) 17-C H ( H) -5.37 (d) 4'-H 5.57 (dxd) 2'-H 5.75 (d) 17-CH ( H ) -7.52 (dxd) 11-H 7.61 (d) 9-H 7.64 ( s) 14-H 8.19 (d) 12-H Example 4 7-Ethylcamptothecin-10-yl β-L-fucopyranoside 1.1 g of 7-ethylcamptothecin-10-yl 2 ', 3', 4'-tri-O-acetyl-β-L-fucopyranoside was suspended in 160 ml of anhydrous methanol, and 0.5 ml of 28% sodium methoxide-methanol solution was suspended in the suspension. Is added and reacted at room temperature for 40 minutes. Post-treatment was carried out in the same manner as in Example 2 and recrystallization from methanol gave 0.77 g of the title compound as colorless prism crystals.

MP207-209 ℃ (decomposition) SIMS: MH ⁺ 539 ¹ H-HMR δ: 400MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.19 (d) 6'-CH ₃ 1.32 (t) 7-CH ₂ CH ₃ 1.78 to 1.96 (m) 20 _{-C H 2 CH 3 3.02~3.16 (m} ) 7-C H (H) CH 3 3.19~3.32 (m) 7-CH (H) CH 3 3.39~3.58 (m) 3'-H, 4'-H 3.61 to 3.70 (m) 2'-H 3.93 (q) 5'-H 4.62 (d) 4'-OH 4.89 (d) 3'-OH 5.15 (d) 1'-H (J ₁ ', ₂ ' = 7.69Hz) 5.27 (d) 2'- OH 5.31 (s) 5-CH 2 - 5.43 (s) 17-CH 2 - 6.51 (s) 20-OH 7.29 (s) 14-H 7.58 (d × d) 11 -H 7.67 (d) 9-H 8.12 (d) 12-H Example 5 3 ', 4', 6'-tri-O-acetyl-1 ', 2'-O-
(7-Ethylcamptothecin-10-ylorthoacetyl) -β-D-mannopyranoside 2.35 g of 7-ethyl-10-hadidoxycamptothecin,
2.90 g of 2,3,4,6-tetra-O-acetyl-α-D-mannopyranosyl bromide and 3.52 g of anhydrous potassium carbonate were suspended in 450 ml of acetone, and the mixture was boiled under reflux for 6 hours with vigorous stirring. Let After completion of the reaction, the insoluble matter is filtered off using Celite, and the filtered material is washed with acetone. The washing solutions are combined and the solvent is distilled off under reduced pressure to obtain a blackish brown tar-like residue. This residue was subjected to medium pressure silica gel column chromatography (chloroform / n-hexane (4/1), chloroform, and then chloroform / methanol (100 /
Then, it was washed with ether and recrystallized from methanol to obtain 0.17 g of the title compound as pale yellow granular crystals.

MP141-144 ℃ SIMS: MH ⁺ 723 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.04 (t) 20-CH ₂ CH ₃ 1.39 (t) 7-CH ₂ CH ₃ 1.83 to 1.96 (m) 20-CH ₂ CH ₃ 1.97, 2.06, 2.10 , 2.13 (s × 4) Orthoacetyl 3 ′, 4 ′, 6′-OCOCH ₃ 3.14 (q) 7-CH ₂ CH ₃ 3.69 (d × d × d) 5′-H 3.86 (s) 20-OH 4.17 (dxd) 6'-C H (H) -4.26 (dxd) 6'-CH ( H ) -4.29 (dxd) 2'-H 5.09 (dxd) 3'-H 5.25 _{(s) 5-CH 2 -} 5.31 (d) 17-C H (H) - 5.33 (d × d) 4'-H 5.46 (d) 1'-H (J 1 ', 2' = 2.56Hz) 5.75 (D) 17-CH ( H ) -7.63 (dxd) 11-H 7.64 (s) 14-H 7.79 (d) 9-H 8.16 (d) 12-H Example 6 7-Ethylcamptothecin-10- IL 2 ', 3', 6 ',
2 ", 3", 4 ", 6" -hepta-O-acetyl-β-D-maltoside 1.57 g of 7-ethyl-10-hydroxycamptothecin and 2.35 g of anhydrous potassium carbonate were suspended in 250 ml of acetone, and the mixture was boiled under reflux with vigorous stirring to give 2,3,6,
50 ml of an acetone solution containing 10.50 g of 2 ', 3', 4 ', 6'-hepta-O-acetyl-α-D-maltolubromide was added dropwise over 30 minutes, and the reaction was carried out for 6 hours. After completion of the reaction, the insoluble matter is filtered off using Celite, and the filtered material is washed with chloroform. After combining the filter washings and distilling off the solvent under reduced pressure, the residue was subjected to medium pressure silica gel column chromatography,
(Chloroform then chloroform / methanol (10
Then, it was recrystallized from ethanol to obtain 1.09 g of the title compound as a yellow powder.

MP157-161 ℃ SIMS: MH ⁺ 1011 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.04 (t) 20-CH ₂ CH ₃ 1.42 (t) 7-CH ₂ CH ₃ 1.83 to 1.96 (m) 20-CH ₂ CH ₃ 2.02,2.04,2.05 , 2.07,2 ′, 3 ′, 6 ′, 2 ″, 3 ″, 2.08,2.09,2.11 (s × 7) 4 ″, 6 ″ -OCOCH ₃ 3.08 to 3.26 (m) 7-CH ₂ CH ₃ 3.82 (S) 20-OH 3.93 to 4.07 (m) 5'-H, 5 "-H 4.08 (dxd) 6" -C H (H) -4.19 (dxd) 4'-H 4.27 (dx d) 6 "-CH (H) - 4.31 (d × d) 6'-C H (H) - 4.55 (d × d) 6'-CH (H) - 4.88 (d × d) 2" -H 5.08 (d × d) 4 "-H 5.20 (d × d) 2'-H 5.26 (s) 5-CH 2 - 5.31 (d) 17-C H (H) - 5.38 (d) 1'-H (J ₁ ', ₂ ' = 7.69Hz) 5.39 (dxd) 3'-H 5.40 (dxd) 3 "-H 5.49 (d) 1" -H (J ₁ ", ₂ " = 4.03Hz) 5.75 ( d) 17-CH ( H ) -7.49 (dxd) 11-H 7.59 (d) 9-H 7.64 (s) 14-H 8.19 (d) 12-H Example 7 7-Ethylcamptothecin-10-yl β-D-maltoside 7-ethylcamptothecin-10-yl 2 ', 3', 6 ',
400 mg of 2 ", 4", 6 "-hepta-O-acetyl-β-D-maltoside was suspended in 45 ml of anhydrous methanol, and 28%
Add 0.2 ml of sodium methoxide-methanol solution,
Incubate for 30 minutes at room temperature. Post-treatment as in Example 2,
Recrystallization from ethanol gave the title compound (269 mg) as colorless needles.

MP220-221 ℃ (decomposition) SIMS: MH ⁺ 717 ¹ H-NMR δ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.32 (t) 7-CH ₂ CH ₃ 1.78 to 1.95 (m) 20-CH ₂ CH ₃ 3.06 to 3.82 _{(m) 7-C H 2} CH 3, 2'-H, 3'-H, 4'-H, 5'-H, 6'-CH 2 -, 2 "-H, 3" -H, 4 " -H, 5 "-H, 6" -CH 2 - 4.53 (t) 6 ' or 6 "-OH 4.68 (t) 6 ' or 6" -OH 4.92 (d) Mal. -OH 4.94 (d) Mal. -OH 5.09 (d) 1 "-H (J 1", 2 "= 3.30Hz) 5.24 (d) 1'-H (J 1 ', 2' = 7.69Hz) 5.32 (s) 5-CH 2 - 5.43 _{(s) 17-CH 2 -} 5.50 (d) Mal.-OH 5.56 (d) Mal.-OH 5.67 (d) Mal.-OH 6.50 (s) 20-OH 7.29 (s) 14-H 7.58 (d × d) 11-H 7.73 (d) 9-H 8.11 (d) 12-H Example 8 Methyl (7-ethylcamptothecin-10-yl 2 ',
3 ', 4'-tri-O-acetyl-β-D-glucopyranoside) -uronate Method A: 7-ethyl-10-hydroxycamptothecin 1.37
g, methyl (2,3,4-tri-O-acetyl-α-D-glucopyranosyl bromide) -uronate (8.34 g) and anhydrous potassium carbonate (2.42 g) were suspended in acetone (300 ml) with vigorous stirring. Boil to reflux for 7 hours. After completion of the reaction, the insoluble matter is filtered off using Celite, and the filtered material is washed with chloroform. After combining the filter washes and distilling off the solvent under reduced pressure, the residue was subjected to medium pressure silica gel column chromatography (eluted with chloroform and then chloroform / methanol (100/1)), and then from ethanol. Recrystallization gave 85 mg of the title compound as colorless needles.

Method B: 7-ethyl-10-hydroxycamptothecin 10.0
g was dissolved in 640 ml of 0.1N potassium hydroxide aqueous solution, and then methyl (2,3,4-tri-O-acetyl-α-D-glucopyranosyl bromide) -uronate 20.0 g of acetone 500
Add ml solution and react at room temperature for 4 hours. Then, the mixture was post-treated in the same manner as in Method B of Example 1 and recrystallized from ethanol to obtain 0.6 g of the title compound as colorless needle crystals.

MP179-182 ℃ (decomposition) SIMS: MH ⁺ 709 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.03 (t) 20-CH ₂ CH ₃ 1.40 (t) 7-CH ₂ CH ₃ 1.81 to 1.97 (m) 20-CH ₂ CH ₃ 2.08, 2.09, 2.12 (S × 3) 2 ′, 3 ′, 4′-OCOCH ₃ 3.14 (q) 7-CH ₂ CH ₃ 3.73 (s) 6′-COOCH ₃ 3.98 (s) 20-OH 4.33 (d) 5′- H 5.24 (s) 5-CH 2 - 5.29 (d) 17-C H (H) - 5.36~5.49 (m) 1'-H, 2'-H, 3'-H, 4'-H 5.73 (d ) 17-CH ( H ) -7.48 (dxd) 11-H 7.61 (d) 9-H 7.61 (s) 14-H 8.13 (d) 12-H Example 9 Methyl (7-ethylcamptothecin-10- IL β-D-
Glucopyranoside) -uronate Methyl (7-ethylcamptothecin-10-yl 2 ',
30 mg of 3 ', 4'-tri-O-acetyl-β-D-glucopyranoside) -uronate was suspended in 3 ml of anhydrous methanol, and a 28% sodium methoxide-methanol solution was suspended therein.
Add 50μ and incubate at room temperature for 30 minutes. Ion-exchange resin (sulfonic acid-H ⁺ type) is added to the reaction solution to make it weakly acidic. The resin is filtered off, washed with methanol, and then the filter washings are combined and dried under reduced pressure. The residue was subjected to medium pressure silica gel column chromatography (eluted with chloroform / methanol (10/1)) and then recrystallized from ethanol to obtain 13 mg of the title compound as colorless to pale yellow amorphous powder.

MP239-241 ℃ (decomposition) SIMS: MH ⁺ 583 ¹ H-NMRδ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.28 (t) 7-CH ₂ CH ₃ 1.78 to 1.95 (m) 20-CH ₂ CH ₃ 3.10 (d × q) 7-C H ( H) CH 3 3.28 (d × q) 7-CH (H) CH 3 3.36~3.54 (m) 2'-H, 3'-H, 4'-H 3.66 (s) _{6'-COOCH 3 4.27 (d)} 5'-H 5.31 (s) 5-CH 2 - 5.34 (d) GlcA.-OH 5.43 (s) 17-CH 2 - 5.46 (d) 1'-H (J 1 ′, ₂ ′ = 7.33Hz) 5.50 (d) GlcA.-OH 5.61 (d) GlcA.-OH 6.50 (s) 20-OH 7.29 (s) 14-H 7.58 (d × d) 11-H 7.68 (d ) 9-H 8.12 (d) 12-H Example 10 7-Ethylcamptothecin-10-yl β-D-glucopyranoside uronic acid Method A: 380 mg of methyl (7-ethylcamptothecin-10-yl 2 ', 3', 4'-tri-O-acetyl-β-D-glucopyranoside) -uronate was suspended in 20 ml of acetone, and 0.1N water was added thereto. Add 32 ml of sodium oxide aqueous solution and react at room temperature for 2.5 hours. After completion of the reaction, pH 4-5 with 1N hydrochloric acid
Acetone was distilled off under reduced pressure at a low temperature, followed by reverse phase medium pressure column chromatography (ODS column) treatment (eluting with water / methanol (7/3)), followed by ethanol- Recrystallized from n-hexane to give compound 28
2 mg was obtained as a pale yellow powder.

Method B: Methyl (7-ethylcamptothecin-10-yl 2 ', 3', 4'-tri-O-acetyl-β-D-glucopyranoside) -uronate 250 mg was suspended in anhydrous methanol 10 ml, and 28% was added thereto. 0.2 ml of sodium methoxide-methanol solution is added, and the mixture is reacted at room temperature for 20 minutes. Then, 0.5 ml of 1N sodium hydroxide aqueous solution was added to the reaction solution,
Incubate for 30 minutes at room temperature. Ion exchange resin (sulfonic acid-H ⁺ acid) is added to the reaction solution to make it weakly acidic. The resin is filtered off, washed with methanol, and then the filter washings are combined and dried under reduced pressure. The residue was treated with reverse phase medium pressure column chromatography (ODS column) (eluted with water / methanol (6/4)) and then recrystallized from ethanol-n-hexane to give 120 mg of the title compound. Obtained as a pale yellow powder.

MP226-229 ℃ (decomposition) SIMS: MH ⁺ 569 ¹ H-NMR δ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.30 (t) 7-CH ₂ CH ₃ 1.78 to 1.95 (m) 20-CH ₂ CH ₃ 3.07 to 3.47 _{(m) 7-C H 2} CH 3, 2'-H, 3'-H, 4'-H 3.76 (d) 5'-H 5.11~5.25 (br-s) GlcA.-OH 5.21 (d) 1 ′ -H (J ₁ ′, ₂ ′ = 7.33Hz) 5.30 (s) 5-CH ₂ − 5.43 (s) 17-CH ₂ −, GlcA.−OH 6.50 (s) 20−OH 7.29 (s) 14− H 7.60 (d × d) 11-H 7.73 (d) 9-H 8.11 (d) 12-H Example 11 7-Ethylcamptothecin-10-yl β-D-glocopyranoside Uronic acid sodium salt 130 mg of 7-ethylcamptothecin-10-yl β-D-glucopyranoside uronitacid is dissolved in 5 ml of distilled water, to which 20 mg of sodium hydrogen carbonate is added to obtain a uniform solution. After filtration and freeze-drying, 134 mg of the title compound was obtained as a yellow powder.

¹ H-NMR δ: 400 MHz (D ₂ O + 1N HCl) 1.01 (t) 20-CH ₂ C H ₃ 1.11 (t) 7-CH ₂ C H ₃ 1.81 to 2.01 (m) 20-CH ₂ CH ₃ 2.49 (d × q) 7-C H ( H) CH 3 2.72 (d × q) 7-CH (H) CH 3 3.62~3.82 (m) 2'-H, 3'-H, 4'-H 3.97 (d) 5-C H (H) - 4.10 (d) 5'-H 4.34 (d) 5-CH (H) - 5.10 (d) 1'-H (J 1 ', 2' = 7.33Hz) 5.26 (d) 17-C H (H) - 5.33 (d) 17-CH (H) - 6.80 (s) 14-H 6.87 (s) 9-H, 11-H 6.98 (s) 12-H eXAMPLE 12 7-ethyl Camptothecin-10-yl 3 ', 4', 6'-tri-O-acetyl-2'-deoxy-2'-trifluoroacetamido-β-D-glucopyranoside 1.96 g of 7-ethyl-10-hydroxycamptothecin
After dissolving in 125 ml of 0.1N potassium hydroxide aqueous solution, 3,4,6
-Tri-O-acetyl-2-deoxy-2-trifluoroacetamide-α-D-glucopyranosyl bromide
A solution of 4.64 g of acetone in 100 ml is added and reacted at room temperature for 2 hours. The yellow substance that precipitates during the reaction is filtered off, the filtered material is washed with acetone, and the filter washings are combined, and the acetone is distilled off under reduced pressure at low temperature. Chloroform and 0.
Add 1N hydrochloric acid and shake. The chloroform layer is separated, washed with water, dried over magnesium sulfate, and concentrated under reduced pressure.
The concentrate is subjected to medium pressure silica gel column chromatography, and the fraction containing the desired product (eluted with chloroform and then chloroform / methanol (100/5)) is collected and dried. The residue was further treated with silica gel column chromatography (chloroform / ether (1/1)
Then, the residue was eluted with chloroform / methanol (10/1) and then recrystallized from chloroform-n-hexane to obtain 80 mg of the title compound as colorless to pale yellow crystalline powder.

MP148-151 ℃ (decomposition) SIMS: MH ⁺ 776 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 0.96 (t) 20-CH ₂ CH ₃ 1.32 (t) 7-CH ₂ CH ₃ 1.79 to 1.93 (m) 20-CH ₂ CH ₃ 2.10,2.11,2.11 (S × 3) 3 ′, 4 ′, 6′-OCOCH ₃ 2.93 (d × q) 7-C H (H) CH ₃ 3.08 (d × q) 7-CH ( H ) CH ₃ 4.04 (a × d × d) 5'-H 4.25 ( d × d) 6'-C H (H) - 4.33 (br) 20-OH 4.37 (d × d) 6'-CH (H) - 4.49 (d × d × d ) 2'-H 4.88 (d) 5-C H (H) - 5.04 (d) 5-CH (H) - 5.26 (d × d) 4'-H 5.27 (d) 17-C H (H) - 5.45 (d) 1'-H 5.47 (dxd) 3'-H 5.71 (d) 17-CH ( H ) -7.35 (dxd) 11-H 7.49 (d) 9-H 7.63 (br) 2 ′ -NHCOCF ₃ 7.65 (s) 14-H 8.08 (d) 12-H Example 13 7-Ethylcamptothecin-10-yl 2′-deoxy-
2'-trifluoroacetamide-β-D-glucopyranoside 7-Ethylcamptothecin-10-yl 3 ', 4', 6'-tri-O-acetyl-2'-deoxy-2'-trifluoroacetamide-β-D-glucopyranoside (65 mg) was dissolved in anhydrous methanol (5 ml). To this, 0.1 ml of 28% sodium methoxide-methanol solution is added, and the mixture is reacted at room temperature for 30 minutes. Ion exchange resin (sulfonic acid-H ⁺
Type) to make the liquid weakly acidic. After removing the resin by filtration and washing with methanol, the washings are combined and dried under reduced pressure. The residue was crystallized from ethanol-n-hexane to give 50 mg of the title compound as a pale yellow powder.

MP204-206 ℃ (decomposition) SIMS: MH ⁺ 650 ¹ H-NMR δ: 40 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.30 (t) 7-CH ₂ CH ₃ 1.78 to 1.95 (m) 20-CH ₂ CH ₃ 3.11 to 3.23 _{(m) 7-C H 2} CH 3 3.24~3.33 (m) 4'-H 3.43~3.62 (m) 3'-H, 5'-H, 6'-C H (H)
-3.78-3.85 (m) 6'-CH ( H ) -3.86-3.95 (m) 2'-H 4.80 (dxd) 6'-OH 5.26 (d) 1'-H (J ₁ ', ₂ = 8.43Hz) 5.31 (d) 3'- OH or 4'-OH 5.32 (s) 5 -CH 2 - 5.42 (s) 17-CH 2 - 5.43 (d) 3'-OH or 4'-OH 6.49 (s ) 20-OH 7.29 (s) 14-H 7.48 (dxd) 11-H 7.70 (d) 9-H 8.13 (d) 12-H 9.37 (d) 2'-NHCOCF ₃ Example 14 7-Ethylcamptothecin -10-yl 2'-amino-
2'-deoxy-β-D-glupyranoside hydrochloride 7-Ethylcamptothecin-10-yl 2'-deoxy-
40 mg of 2'-trifluoroacetamide-β-D-glucopyranoside is suspended in 4 ml of water, and 1.85 ml of 0.1N sodium hydroxide aqueous solution is added thereto, and the mixture is reacted at room temperature for 2.5 hours.
The reaction mixture was ice-cooled, 0.1 ml of hydrochloric acid (3 ml) was added to the mixture to acidify it, and then medium pressure reverse phase column chromatography (ODS column)
Workup (eluted with water / methanol (7/3 then 1/1)) then crystallized from methanol-n-hexane to give 24 mg of the title compound as a pale yellow powder.

MP223 to 226 ℃ (decomposition) SIMS: MH ⁺ 554 ¹ H-NMR δ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.32 (t) 7-CH ₂ CH ₃ 1.78 to 1.96 (m) 20-CH ₂ CH ₃ 2.86 (t -like) 2'-H 3.18 (q ) 7-C H 2 CH 3 3.22~3.38 (m) 3'-H, 4'-H 3.46~3.79 (m) 5'-H, 6'-C H ( H) - 3.73~3.83 (m) 6' -CH (H) - 4.71 (d × d) 6'-OH 5.11 (d) 1'-H (J 1 ', 2' = 8.06Hz) 5.15~5.22 ( br) 3'-OH or 4'-OH 5.24~5.54 (br) 3' -OH or _{4'-OH, 2'-NH 2} · HCl 5.31 (s) 5-CH 2 - 5.43 (s) 17-CH _{2 - 6.50 (s) 20-} OH 7.29 (s) 14-H 7.62 (d × d) 11-H 7.78 (d) 9-H 8.12 (d) 12-H eXAMPLE 15 camptothecin-10-yl 2 ', 3 ', 4', 6'-Tetra-
O-acetyl-β-D-glucopyranoside 900 mg of 10-haladoxycamptothecin was dissolved in 68 ml of 0.1N potassium hydroxide aqueous solution, and then 2,3,4,6-tetra-
O-acetyl-α-D-glucopyranosyl bromide 2.
A solution of 24 g of acetone in 60 ml is added, and the mixture is reacted at room temperature for 2 hours. After filtering off the yellow substance that precipitates during the reaction, acetone is distilled off under reduced pressure at a low temperature. Chloroform and 0.1N hydrochloric acid are added to the resulting aqueous solution and shaken. The chloroform layer is separated, washed with water, dried over magnesium sulfate, and concentrated under reduced pressure.

The concentrate was subjected to medium pressure silica gel column chromatography (eluted with chloroform and then with chloroform / methanol (100/4)) and then crystallized from chloroform-n-hexane to obtain 75 mg of the title compound as a colorless powder. .

MP268-269 ℃ (decomposition) SIMS: MH ⁺ 695 ¹ H-NMR δ: 400 MHz (CDCl ₃ ) 1.04 (t) 20-CH ₂ CH ₃ 1.83 to 1.97 (m) 20-CH ₂ CH ₃ 2.07,2.08,2.10 (s × 3) 2 ′, 3 ′, 4 ', 6'-OCOC
_{H 3 (2.08 × 2) 3.79} (s) 20-OH 3.98 (d × d × d) 5'-H 4.23 (d × d) 6'-C H (H) - 4.34 (d × d) 6'- CH (H) - 5.20~5.43 (m ) 5-CH 2 -, 17-C H (H) -, 1'-H, 2'-H, 3'-H, 4'-H 5.74 (d) 17 -CH ( H ) -7.40 (d) 9-H 7.52 (dxd) 11-H 7.64 (s) 14-H 8.18 (d) 12-H 8.27 (s) 7-H Example 16 Camptothecin-10- Yl β-D-glucopyranoside Camptothecin-10-yl 2 ', 3', 4 ', 6'-tetra-
50 mg of O-acetyl-β-D-glucopyranoside is suspended in 20 ml of anhydrous methanol, 0.1 ml of 28% sodium methoxide-methanol solution is added, and the mixture is reacted at room temperature for 30 minutes. Ion-exchange resin (sulfonic acid-H ⁺ type) is added to the reaction solution to make it weakly acidic. The resin is filtered off, washed with methanol, and then the filter washings are combined and dried under reduced pressure. The residue was recrystallized from ethanol to give 33 mg of the title compound as colorless ~
Obtained as pale yellow needles.

MP214-216 ℃ (decomposition) SIMS: MH ⁺ 527 ¹ H-NMR δ: 400 MHz (DMSO-d ₆ ) 0.88 (t) 20-CH ₂ CH ₃ 1.78 to 1.98 (m) 20-CH ₂ CH ₃ 3.18 to 3.28 (m) 4'-H 3.30 to 3.39 ( m) 2'-H, 3'-H 3.42 to 3.58 (m) 5'-H, 6'-C H (H) -3.69 to 3.81 (m) 6'-CH ( H ) -4.60 (d x d ) 6'-OH 5.07 (d) 4'-OH 5.12 (d) 1'-H (J ₁ ', ₂ ' = 6.96Hz) 5.14 (d) 2'-OH or 3'-OH 5.28 (s) 5 _{-CH 2 - 5.42 (s) 17} -CH 2 -, 2'-OH or 3'-OH 6.50 (s) 20 -OH 7.30 (s) 14-H 7.59 (d × d) 11-H 7.65 (d) 9-H 8.11 (d) 12-H 8.55 (s) 7-H The present invention provides a novel compound having a higher activity than camptothecin. The compound provided by the present invention, or a salt thereof is as described below. It has excellent antitumor activity and is considered to be used as an anticancer agent.

The antitumor activity of the compounds provided by the invention is
An example will be described below.

Experimental Example 1 Antitumor effect on mouse leukemia L1210 Experimental method 5 × 10 ⁵ mouse leukemia L1210 was 7 weeks old (body weight 17 to 19).
Female CDF ₁ mice (g) (6 per group) were intraperitoneally transplanted, and the test substance was intraperitoneally administered 1, 5 and 9 days after the transplantation, and the life prolonging effect was observed.

The test substance was dissolved or suspended in physiological saline and administered.
The total dose was as shown in Table 1. The antitumor effect was obtained by multiplying the ratio of the average survival time (T) of the drug-administered group and the average survival time (C) of the non-drug-administered group by 100 times under the above experimental conditions (T / C).
%).

Experimental Results 7-Ethylcamptothecin-10-yl 2 ′, 3 ′, 4 ′, 6′-tetra-O-acetyl-β-D-glucopyranoside (Compound 1) and 7-ethylcamptothecin-10-obtained in the examples. Table 1 shows the results of an antitumor effect test conducted on yl β-D-glucopyranoside (Compound 2). For comparison, the antitumor effect of 7-ethylcamptothecin sodium salt (Compound 3) was shown.

As can be seen from the experimental results described above, the novel glycoside derivative of camptothecin provided by the present invention exhibits excellent antitumor activity against experimental animal tumors.

Further, among the novel glycoside derivatives of camptothecin obtained by the present invention, the compound in which the protecting group in the sugar moiety is removed has excellent water solubility as compared with camptothecin, 7-lower alkylcamptothecin and their hydroxy-substituted derivatives. .

The compound provided by the present invention or a salt thereof has excellent antitumor activity and excellent water solubility, and can be used as an anti-malignant tumor therapeutic agent.

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor, Matahiko Hodai, 1-1-19 Higashi-Shimbashi, Minato-ku, Tokyo Yakult Honsha Co., Ltd. (72) Inventor, Tomio Furuta 1-1, Higashi-Shimbashi, Minato-ku, Tokyo No. 19 Yakult Honsha Co., Ltd. (72) Inventor Teruo Yokokura 1-1-19 Higashishimbashi, Minato-ku, Tokyo Yakult Honsha Co., Ltd.

Claims (1)

[Claims] 1. A general formula (I) (In the formula, R ¹ represents a hydrogen atom or a C _1-4 alkyl group, R ² is an aldose residue, or —COOH group, —NH ₂ group or —OH group present in the group is Aldose protected by a lower aliphatic acyl group having 1 to 2 carbon atoms in the alkyl group, a benzoyl group, a trifluoroacetyl group, a lower alkyl ester group having 1 to 2 carbon atoms in the alkyl group, or a combination thereof. A camptothecin derivative or a salt thereof, which represents a residue, wherein the bond between R ² and camptothecin represents a glycosidic bond between the oxygen atom on the A ring of camptothecin and the 1st carbon atom of the sugar.