JPH01313426A - Saccharide lactam derivative and cancerous cell metastatic inhibitor containing the same - Google Patents

Abstract

NEW MATERIAL:A compound expressed by formula I [R<1> and R<2> are H, formula II, III, etc., (R<3> to R<8> are H, halogen, lower alkyl, lower alkoxy, etc.; R<9> is lower alkyl, etc.); W is H or acetyl, except when all the R<1>, R<2> and W are H]. EXAMPLE:3-O-Trityl-D-gluco-delta-lactam. USE:A cancer cell metastatic inhibitor capable of remarkably inhibiting metastasis of cancerous cells with extremely low cytotoxicity. PREPARATION:A D-gluco-delta-lactam expressed by formula IV is reacted with an activated trityl compound expressed by formula V (X is halogen) in the presence of a base (e.g., triethylamine) in a solvent, such as N,N- dimethylformamide, at 5-100 deg.C for 1-48hr to afford the compound expressed by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a novel glycolactam derivative that significantly inhibits the formation of metastatic foci of cancer cells, and a cancer cell metastasis inhibitor containing the same.

[Conventional technology]

Conventional anticancer drugs are mainly drugs that kill malignant cells through the cell-killing properties of substances or via the human immune system.

These drugs are still not sufficient for the treatment of cancer. In addition, solid cancers are treated with surgery and radiation therapy, and the success rate of removing the primary cancer has greatly improved. In fact, the most important factor that affects the prognosis of cancer patients is cancer metastasis.

For these reasons, it is expected that the effectiveness of the various current therapies will be further enhanced by suppressing the metastasis of cancer cells. None of them are in full-scale use.

[Problem to be solved by the invention]

The present inventors conducted research on drugs mainly having an anti-metastatic effect, especially substances that have an effect of inhibiting cancer cell metastasis, and first discovered a glycolactam derivative that has an effect of inhibiting cancer cell metastasis. I applied on the day.

An object of the present invention is to provide novel D-gluulose delta-lactam derivatives and anticancer agents based on their cancer cell metastasis inhibiting action.

[Means to solve the problem]

The present invention is based on the general formula [wherein R' and R2 may be the same or different hydrogen atoms (wherein R3 to R8 may be the same or different hydrogen atoms, halogen atoms, lower alkyl groups, lower alkokene groups, Halogen methyl group, carboxyl group.

represents a carboxyalkyl group or a carbalkoxyalkyl group, R9 represents a lower alkyl group or (in the formula, R5 and R6 have the same meanings as above), and w represents a hydrogen atom or an acetyl group. However, the case where R1゜R2, W are all hydrogen atoms is excluded] and the cancer cell metastasis inhibitor which contains the same sugar lactam derivative as an active ingredient.

The sugar lactam derivative represented by the general formula (1) of the present invention is
Nojirimaine (5-amino-
5-deoxy-D-glucobylanose) (JP-A-Sho, 13
-760 Publication, Tetrahedron, Volume 23,
2125, 1968)) (Meiji Seika Research Annual Report).

Volume 13, pp. 80-84, 1973, Special Publication 1973-2
8375) as a raw material, it can be obtained by the following manufacturing process.

R of the compound (1) of the present invention represented by the above formula (1)
Compound (Ia) in which 1°R2 is (R3-R8 have the same meanings as above) can be produced by the following reaction formula (la).

(In the formula, X represents a halogen atom, and R3-R6 have the same meanings as above.) In this reaction, N,N-dimethylformamide, N.

In an inert polar solvent such as N-dimethylacetamide, dimethylsulfoxide, pyridine, an inorganic base such as potassium carbonate, sodium carbonate, sodium hydride, barium oxide, hydrogen carbonate, or triethylamine, pyridine, diisopropylethylamine, Synchhexylamine, 1゜5-diazabicyclo(4,3,0
) D-glucose in the presence of an organic base such as non-5-ene
It is obtained by reacting with δ-lactam (↓) and activated trityl compound (7), usually at 5 to 100°C for 1 to 48 hours.

R1 of the compound (1) of the present invention represented by the above formula (1)
Compound (Ib) in which ゜R2 is \ (wherein R', R', R9 have the same meanings as above) can be prepared by combining \ and D-gluco-δ according to the production of (Ia) above. -It can be obtained by reacting with lactam (yo).

However, the reaction temperature is generally high, e.g.
C. or the 3- or 6-position of D-gluco-delta-lactam, the reaction using the following diazo compound is simple.

Ren (wherein RIO is a hydrogen atom or an acyl protecting group, R3°R' and R9 have the same meanings as above) This reaction uses methylene chloride, 1,2-dichloroethane, tetrahydrofuran, dioxane, and acetone.

The reaction is carried out in an inert solvent such as ethyl acetate, toluene, acetonitrile, N,N-dimethylformamide, or a mixed solvent thereof, usually in the presence of an acid catalyst.

The acid catalysts used are tetrafluoroboric acid and perchloric acid.

Examples include protons such as trifluoromethanesulfonic acid, tetrafluoroboric acid, silver tetrachloride, ferric chloride, antimony pentafluoride, and zinc chloride.

The (Ib) type compound of the compound (+) of the present invention represented by the above formula (1) can be obtained at a reaction temperature of -60 DEG to +50 DEG C. and a reaction time of 1 to 24 hours.

Compounds of the present invention include, but are not limited to, the sugar cucum derivatives shown below.

3-0-) Lichiru D-Gullo δ-Lakucum.

6-0-) Lythyl-D-glucose δ-lactam.

2-0-) Lythyl-D-glucose δ-lactam.

3.6-di-0-trityl-D-gluco-δ-lactam, 6-0-(3,5-ditrifluoromethyl-trityl)-D-gluco-δ-lactam, 6-0-(3,3° −
ditrifluoromethyl-trityl)-D-gluco-δ-
lactam, 5-Q-(4-tert-butyl-trityl)-D-gluco-δ-lactam.

6-O-(4-carboxy-trityl)-D-gluco-
δ-lactam, 6-0-benzhydryl-D-gluco-
δ-lactam, 6-0-(3,5-cyto, trifluoromethyl-benzhydryl)-D-gluco-δ-lactam, 6-0-(3,3°-ditrifluoromethyl-benzhydryl)-D-gluco -δ-lactam, 6-O-(
1-β-naphthyl-ethyl)-D-gluco-δ-lactam, 6-O-(p-phenyl-benzhydryl)-D-
Gluco-δ-lactam, 6-0-(1-phenyl-ethyl)-D-gluco-δ-lactam, 6-0-(α-chloro-5-nitro-benzhydryl)-D-gluco-δ
-lactam, 6-0-(3,4-dichloro-benzhydryl)-D-gluco-δ-lactam, 6-0-(4,
4°-dichloro-benzhydryl)-D-gluco-δ-
lactam, 6-0-(2,4-difluoro-benzhydryl)-D-gluco-δ-lactam, 6-0-(4-nitro-benzhydryl)-D-gluco-δ-lactam,
6-0-(1-p-phenoxy-ethyl)-D-
'j glucololactam, 6-0-(3,4-dimethyl-benzhydryl)-D-gluco-δ-lactam,
6-0-m-(1-tert-butyloxycarbonyl-ethyl-benzhydryl)-D-gluco-δ-lactam.

Next, production examples of the compounds of the present invention will be shown.

[Production Example] Production Example 1 Hydrazone compound 1 synthesized from 6-0-(3,5-citrifluoromethyl-benzhydryl)-D-gluco-δ-lactam 3.5 to ditrifluoromethylbenzophenone according to a conventional method ．． Og was dissolved in 20 ml of ethyl acetate, nickel peroquinide 3 and Og were added, and the mixture was stirred at room temperature for 1 hour.

After removing the insoluble portion, ethyl acetate was distilled off at an external temperature of 30° C. or lower to obtain 0.95 g of a diazo compound.

095 g of the obtained diazo compound and 2,3.1-)
Acetyl-D-glucose δ-lactam 500 mg 1
, 2-dichloroethane (10 m2) and cooled to 5°C. While stirring, in an argon stream, a solution of 40 mg of tetrafluoroboric acid etherate in 1 m (l) of 1,2-dichloroethane was slowly added dropwise. After the dropping was complete, the mixture was stirred at 5°C for 30 minutes. The liquid obtained by concentration The product was purified by silica gel column chromatography (toluene-ethyl acetate 2:I) to obtain 2.3.4-triO-acetyl-6-0-(3,
5-ditrifluoromethylbenzhydryl)-D-gluco-δ-lactam [Silica gel thin layer chromatography (toluene-ethyl acetate 1) Rt Q, 34E
540 mg was obtained.

270 mg of the 2,3,4-trilow acetyl protected 6-0-substituted product obtained above was dissolved in 4 d of methanol, and a freshly prepared methanol solution of sodium ions was added at any time, followed by stirring at room temperature.

The reaction was checked by silica gel thin layer chromatography (chloroform-methanol 4:1), and after the reaction was completed, the same amount of water as the reaction solution was added, treated with Amberlyst 15 to remove sodium ions, and then concentrated to dryness. did. Five portions of diethyl ether were added to the residue, and the mixture was stirred at room temperature for 30 minutes. The insoluble solid part was collected by filtration, and the title compound [silica gel thin layer chromatography (40 form-methanol 4
:1) Rro, 61 mass spectrometry (F, D, rrIa
ss) m/z +479(M+) 1 150
mg was obtained.

Production Example 2 6-0-<3.3"-ditrifluoromethylbenzhydryl)-D-gluco-δ-lactam- Book - Substitute for the hydrazone form of 3.5-ditrifluoromethylbenzhydryl in Production Example 1 Production Example 1
In the same manner as above, the title compound [silica gel thin layer chromatography (chloroform-methanol 4': 1)
Rr O,51, mass spectrometry (FD, mass) m
/z:480(M')] was obtained.

! ! ! Preparation Example 3 6-0-p-phenylbenzhydryl-D-gluco-δ
- Lactam Production Example 1 The title compound [Silica gel thin layer chromatograph Fi (chloroform-methanol 4: 1) Rr O, 64, mass spectrometry (F, D, mass) m/z: 419 (
M")) was obtained.

Production example 4 6-O-(1-β-naphthyl-ethyl)-D-gluco-
The title compound [silica gel thin layer] was prepared in the same manner as in Production Example 1, using 1-β-naphthyl-ethyl hydrazone instead of the hydrazone of 3,5-ditrifluoromethylbenzhydryl in δ-lactam Production Example 1. Chromatography (chloroform-methanol 4:1) R, 0,4, mass fraction tJ?
(F, D, mass) m/z:331(
M") :] was obtained.

Production Example 5 6-O-(1(3,4-dimethoxyphenyl)-ethyl 1-D-gluco-δ-lactam Instead of the hydrazone form of 3.5-ditrifluoromethylbenzhydryl in Production Example 1, 1 Production Example 1 using the hydrazone form of -(3,4-ristoquinphenyl)-ethyl
The title compound [silica gel thin layer chromatography (chloroform-methanol 4:1) R,
0,43 mass spectrometry (F, D.

mass) m/z: 341 (M")] was obtained.

Production Example 6 6-0-(3,4-dichlorobenzhydryl)-D-gluco-δ-lactam Instead of the hydrazone form of 3,5-ditrifluoromethylbenzhydryl in Production Example 1, 3,4- Using the hydrazone compound of dichlorobenzhydryl, the title compound was prepared in the same manner as in Production Example 1 [Silica gel thin layer chromatography (chloroform-methanol'4: l) R,,0,46
, mass spectrometry (F, D, mass) m/z:
411(M”):1 was obtained.

Production Example 7 3.6-di-0-trityl-D-gluco-δ-lactam 1.7 g of D-gluco-δ-lactam and 5.6 g of trityl chloride were mixed with 50 rn1. and heated and stirred at 50° C. overnight. After pyridine was distilled off, chloroform and saturated aqueous sodium bicarbonate were added, and the mixture was extracted three times with chloroform. After drying the chloroform layer with anhydrous potassium carbonate, chloroform was distilled off, and then purified by silica gel thin layer chromatography (chloroform-methanol 70:1) to obtain the title compound [silica gel thin layer chromatography (chloroform-methanol 50:1) R,0
, 25, mass spectrometry (F, D.

mass) m/z:660(M”) ] 0
．． 8g was obtained.

Production Example 8 6-0-(3,3'-ditrifluoromethyltrityl)
-D-Gluco-δ-lactam 1.06 g of D-gluco-δ-lactam and 3.0 g of 3,3''-ditrifluoromethyltrityl chloride were dissolved in 40 ml of pyridine, and the mixture was heated and stirred at 60°C overnight. After distilling off, add ethyl acetate and saturated sodium bicarbonate solution,
The aqueous layer was further saturated with IJium chloride and extracted three times with ethyl acetate. After drying the ethyl acetate layer over anhydrous potassium carbonate, the solvent was distilled off and subjected to silica gel column chromatography (chloroform-methanol 20:1).
The title compound was purified by silica gel thin layer chromatography (chromatography, loform-methanol 10:1).
600 mg of R,0,23) was obtained.

Production Example 9 6-0-(3,5-ditrifluoromethyltrityl)-
The title compound [silica gel thin layer] was prepared in the same manner as in Production Example 3, except that 3,5-ditrifluoromethyltrityl chloride was used instead of 3,3°-ditrifluoromethyltrityl chloride in D-gluco-δ-lactam Production Example 8. Chromatography (chloroform-
Methanol 4:1) RrO053, mass spectrometry (
F, D, mass) m/z: 555 (M
'>II was obtained.

Production Example 10 5-Q-(4-tert-butyltrityl 1-D-gluco-δ-lactam 4-tert-butyltrityl chloride was used instead of 3,3°-ditrifluoromethyltrityl chloride in Production Example 8. , The title compound was obtained in the same manner as in Production Example 8 [silica gel thin layer chromatography (chloroform-methanol 6:1) R,0,38, mass spectrometry (F,
D, mass) m/z: 475 (M+)) was obtained.

The cancer cell metastasis inhibitor of the present invention is clinically administered intravenously, arterially, cutaneously, and subcutaneously as an oral or parenteral preparation containing the above-mentioned 〇-substituted-D-glucose δ-lactam.

It is administered intradermally, rectally, intramuscularly, or orally.

Moreover, stronger effects can be expected by administering directly to the tumor. The dosage varies depending on the dosage form, dosage form, age, weight, and disease state of the patient, but is approximately 100 to 300 doses per day.
Administer 0 mg once or several times.

Parenteral preparations include sterile aqueous or non-aqueous solutions or emulsions. Bases for non-aqueous solutions or emulsions include propylene glycol, polyethylene glycol, glycerin, olive oil, corn oil,
Examples include ethyl oleate.

Oral preparations include capsules, granules, fine granules, powders, and the like.

These preparations contain starch and lactose as excipients.

Contains mannitol, ethylcellulose, sodium chloride, methylcellulose, etc. Magnesium stearate or calcium stearate is added as a lubricant. As the binder, gelatin, gum arabic, cellulose ester, polyvinylpyrrolidone, etc. are used.

It should be noted that acute toxicity (L
Dso value) is 0.3/l:g or more after 8 mouths of administration.

Next, formulation examples and effects of the present invention will be explained.

〔Example〕

Example 1 6"0-) 'J phenylmethyl-D-glucoro-
Lactam 200mg lactose
130mg potato starch
70mg polyvinylpyrrolidone
Mix 10mg magnesium stearate, 2.5mg lactose and potato starch, add a 20% ethanol solution of polyvinylpyrrolidone, moisten the mixture uniformly, pass through a sieve with a mesh size of 1 [1 + 11], dry at 45°C, and dry again. It was passed through a 1 m+n mesh sieve. The granules thus obtained were mixed with magnesium stearate and formed into tablets.

Example 2 6-0-(3,3'-ditrifluoromethyltrityl)
-D-gluco-δ~lactam is ground in a pin mill to particles of approximately 10 μm, and 500 mg thereof is filled into an injection vial to prepare an injection.

When administering this injection, TWEEN
5ml of a solution of 5mg of 80 and 10mg of Nissho Pharmaceutical gelatin dissolved in 5me of distilled water for injection! Add to the injection, shake well and administer the appropriate amount intramuscularly to form a suspension.

Anticancer Cell Metastasis Suppression Effect Test Test Method Fiddler from mouse tumor melanoma 816 strain
(Fiddler) Method in Can
cerRe5erch.

15, pp. 1399-439, 1978) 81
6 highly metastatic strains were selected and used, and the metastatic effect was evaluated by the method of Kijima-3ucla et al. (Cancer Research, Vol. 46, 858-8).
62, 1986).

First, the B16 highly metastatic strain was prepared using Dulbecco M with fetal bovine serum added.
Cells were grown in E medium (DME medium), added with O-substituted-D-glucololactam of the general formula (1) of the present invention, and cultured at 37°C in the presence of 5% CO2 for 2 to 4 days to proliferate the cells. was detached from the culture vessel using trypsin-EDTA solution.

These cells were treated as living cells with Dulbecco's balanced salt solution that does not contain Ca″1 and Mg″ so that the number of I XIO′ cells per l me! 8 It became cloudy.

0.1 mg of this suspension was injected into the mouse tail vein, and
After rearing for 4 days, the lungs were removed through laparotomy, and metastatic nodules of the B16 highly metastatic strain formed on and inside the lungs were counted and compared with a control that was not treated with the drug.

Test Example 1 Cytotoxicity of 6-0-triphenylmethyl-D-gluco-δ-lactam B16 highly metastatic strain and mouse-derived tumor cell L929 strain were incubated in a DME medium supplemented with 10% tallow serum with 5% CO7 After culturing at 37°C in the presence of
The highly metastatic strain is IX104 cells, and the L929 strain is IXIO cells.
' Cells were suspended in the above medium until they became .

150I of this suspension was added to N-(6-0-)liphenylmethyl)-D-gluco-δ-lactam or Adriamain (control) solution 504 and mixed. The 816 highly metastatic strain was cultured for 3 days, and the L929 strain was cultured for 4 days, and the viability was observed under an inverted microscope to determine cytotoxicity. The results were as shown in Table 1.

(The following is the margin of this page) Table 1 In the table, 10 indicates growth and - indicates death.

As is clear from the above test results, 6-0-) is the active ingredient of the present invention! J phenylmethyl-D-glucoro-
Lactam is effective against B16 highly metastatic strain and L929 strain.
No significant cell damage was shown even at 0 K/ml.

Test Example 2 Anti-metastatic effect of 6-0-triphenylmethyl-D-gluco-δ-lactam A B16 highly metastatic strain was planted in a DME medium supplemented with 10% tallow serum, and 6-0-)! 1 mj of I-phenylmethyl-D-curquo-lactam! Add 30g per bottle, 5% CO2
The cells were cultured for 3 days at 37°C in the presence of.

The cells were peeled off from the culture container in the same manner as in Test Example 1, and 1
C so that live cells per ml are IXl06 cells.
The cells were suspended in Dulbecco's balanced salt solution containing no a++ and Mg, and 0.1 yd of the suspension was injected into the tail vein of MDFl mice (8 weeks old, male), and the cells were transplanted. After 14 days of rearing and observation, laparotomy was performed. The lungs were removed and the metastatic nodules of the BI6 highly metastatic strain formed on and inside the lungs were counted.
Shown in the table.

As is clear from the test results in Table 2 and above, 6-0-)U phenylmethyl-, which is the active ingredient of the cancer metastasis inhibitor of the present invention,
D-gluco-δ-lactam significantly inhibited the metastasis of the B16 highly metastatic strain to the lungs.

〔Effect of the invention〕

The novel lactam derivative of the present invention is a useful compound that inhibits the metastasis of cancer cells, and the cancer cell metastasis inhibitor of the present invention containing this compound as an active ingredient inhibits the metastasis of cancer cells very significantly, and This drug has very little cytotoxicity.

Therefore, it is a useful drug that can significantly enhance its anticancer effect even when administered simultaneously with existing anticancer drugs, or when used during surgical therapy or radiotherapy.

Claims (1)

[Claims] 1. General formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, R^1 and R^2 may be the same or different hydrogen atoms, ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ , ▲Mathematical formula, chemical formula,
There are tables, etc. ▼ or ▲ There are mathematical formulas, chemical formulas, tables, etc. , represents a carboxyl group, a carboxyalkyl group, a carbalkoxyalkyl group, and R^9 is a lower alkyl group or ▲There are numerical formulas, chemical formulas, tables, etc.▼ (In the formula, R^5 and R^6 have the same meanings as above). represent}, W
represents a hydrogen atom or an acetyl group. However, R^1, R
A sugar lactam derivative represented by ^2, except when all Ws are hydrogen atoms. 2. A cancer cell metastasis inhibitor comprising the glycolactam derivative according to claim 1 as an active ingredient.