CN1267444C

CN1267444C - Compound thiophosphates containing selenium with fluorin being added to and synthesis

Info

Publication number: CN1267444C
Application number: CN 200410047109
Authority: CN
Inventors: 许新华; 张秋林; 李言杰; 陈雄; 张青丽
Original assignee: Hunan University
Current assignee: Hunan University
Priority date: 2004-12-28
Filing date: 2004-12-28
Publication date: 2006-08-02
Anticipated expiration: 2024-12-28
Also published as: CN1660873A

Abstract

The present invention relates to a seleniferous thiophosphate compound of tegafur and synthesis thereof. The seleniferous thiophosphate compound of tegafur relates to an organic selenium and a nucleoside type substance. The structure of the compound is disclosed in a general formula. The compound is synthesized by two-step pan boiling, and anhydrous benzene is used as a solvent. Below 65 DEG C, iodine powder is used as a catalyst to cause 2-N, N-diethylamino-1, 3, 2-phospho hetero pentane to react with hydroxyalkyl tegafur and the sulphur powder in sequence, in this way, an intermediate body (2) is synthesized. Then, diaryl diselenide is reduced by sodium borohydride to obtain diaryl sodium selenide which carries out nucleophilic ring opening to the intermediate body (2) at room temperature and anhydrous alcohol. The seleniferous thiophosphate compound of tegafur with high yield is formed. By the detection of antineoplastic activity, the compound of the present invention has good inhibiting effect for the bladder cancer cells and the gastric carcinoma cells of human bodies and has little toxicity to epithelial cells. Thus, the introduction of selenium is one of efficient paths to improve the activity of the phospholipid type compound of tegafur.

Description

Contain the thiophosphatephosphorothioate compounds of Tegafur of selenium and synthetic

Technical field: a kind of thiophosphatephosphorothioate compounds of the Tegafur that contains selenium and synthetic, relate to organoselenium and a kind of nucleosides material, specifically be the phosphorothioate derivative that contains the Tegafur of selenium.

Background technology: phosphatide has physiologically active widely, and a large amount of phosphatide has been synthesized and has proved to have good anti-cancer activity, and it is again the effective carrier of medicine simultaneously, and tumor tissues is had certain targeting; Nucleosides and analogue thereof are widely used as antitumor and antiviral clinically; Selenium is that human body must be micro-, it has very significantly anti-cancer, antitumous effect, clinical effectiveness shows, organic selenium can improve very effectively selenium bioavailability and as life must the trace element physiological function, and its toxicity reduces greatly than inorganic selenium, its compound is being brought into play great function in treatment aspect the cancer, in recent years, Se is incorporated in the drug molecule or synthetic new contains the focus that the Se medicine is a research.But selenium is induced one except that this seminar has carried out some preliminary researchs, to yet there are no bibliographical information in the phosphatide nucleosides conjugate.We design two kinds of methods selenium are induced one in the phosphatide nucleosides conjugate, and the first will be connected with the three valent phosphors selenizing of nucleosides, and it two is cyclic glycerol phospholipid conjugates of synthetic nucleosides, by the organoselenium group it is carried out nucleophilic ring opening subsequently.Recently, the selenium (sulphur) that this seminar has finished Tegafur is for synthetic and selenium (sulphur) phenol of the cyclic glycerol phospholipid conjugates nucleophilic ring opening to it, the phospholipid derivative that synthesizes the novel Tegafur of several class formations, external activity experiment shows, to the inhibition effect of human bladder's cancer cells than the good (patent No.: the ZL 02 139741.4 of former medicine Tegafur; Application number: 200410046927.5,200410046925.6,200410046926.0).On this basis, carry on the novel synthetic and activity research that contains selenium nucleosides phosphatide (ester) conjugate, will have certain directive significance the initiative of phospholipid new drug.

Summary of the invention: the objective of the invention is to the sulfo-phosphide compounds of the Tegafur that contains selenium of synthesizing new, low in the hope of solving the nucleoside medicine curative effect, the big deficiency that waits of toxicity.

Realize that technical scheme of the present invention is: compound of the present invention is a kind of thiophosphatephosphorothioate compounds of the Tegafur that contains selenium shown in general formula:

Ar＝C ₆H ₅，p-CH ₃C ₆H ₄，m-CH ₃C ₆H ₄

Its synthetic method is:

Ar＝C ₆H ₅，p-CH ₃C ₆H ₄，m-CH ₃C ₆H ₄

Be described in further detail the present invention below:

Synthetic contains the thiophosphatephosphorothioate compounds of the Tegafur of selenium ³¹PNMR (85%H ₃PO ₄Be external standard) to measure with BRUKER AC-P400HE type instrument, mass spectrum is measured by Agilent 1100 Series, and fusing point is to measure by capillary tube technique. ³¹PNMR, MS and physicochemical data see Table 1.

Table 1.3a～3c's ³¹P NMR and physicochemical data

Entry	R	Molecular formula	R _f	Fusing point (℃)	³¹P NMR (ppm)	MS (M+1-Na)	Yield (％)
Entry	R	Molecular formula	R _f	Fusing point (℃)	³¹P NMR (ppm)	MS (M+1-Na)	Yield (％)	3a	C ₆H ₅	C ₁₈H ₂₁FN ₂O ₆PSSeNa	0.32	87～88	59.166	523.1	92
3b	p-CH ₃C ₆H ₄	C ₁₉H ₂₃FN ₂O ₆PSSeNa	0.37	80～81	58.769	537.2	83	3a	C ₆H ₅	C ₁₈H ₂₁FN ₂O ₆PSSeNa	0.32	87～88	59.166	523.1	92
3b	p-CH ₃C ₆H ₄	C ₁₉H ₂₃FN ₂O ₆PSSeNa	0.37	80～81	58.769	537.2	83	3c	m-CH ₃C ₆H ₄	C ₁₉H ₂₃FN ₂O ₆PSSeNa	0.38	72～73	59.523	537.2	86

Illustrate: measure 3a～c Rf value R _fUsed moving phase is methyl alcohol/trichloromethane (1: 5); Yield is the productive rate behind the column chromatography.

Measure with BRUKER AC-P400HE type instrument ¹H NMR (is interior mark with TMS) and the results are shown in Table 2.

The real spectrum of table 2.2a～3c hydrogen nuclear magnetic resonance ¹H NMR is as follows:

3a：7.193-7.480(m，6H)；6.016(s，1H)；3.933-4.313(m，8H)；3.078(t，2H， ³J _H-H＝7.2Hz)；1.829-2.436(m，4H).
	3b：7.255-7.342(m，3H)，7.013-7.082(m，2H)，6.013(s，1H)；3.936-4.319(m， 8H)；3.083(t，2H， ³J _H-H＝7.2Hz)；2.273(s，3H)，1.831-2.438(m，4H).
3c 7.063-7.532(m，5H)；6.012(s，1H)；3.934-4.312(m，8H)；3.079(t，2H， ³J _H-H＝7.2Hz)；2.301(s，3H)，1.836-2.445(m，4H).

Adopt international tetrazolium (MTT) colorimetry, experiment in vitro has been measured compound 3a～c human body has been reached normal liver epithelial cell L-02 toxic action effect transitional cell bladder carcinoma cell line T-24 and stomach cancer cell BGC-823 restraining effect, and T-24, BGC-823 and the L-02 cell strain of in-vitro cultivation selected in this test for use.Cell doubling time be 24～30h. its results are shown in Table 3.

(half is put dead amount ID to table 3 compound 3a～c to transitional cell bladder carcinoma cell line T-24, stomach cancer cell BGC-823 restraining effect and to the toxic action of normal liver epithelial cell L-02 ₅₀, μ g/m1)

Entry	T-24	BGC-823	L-02
Entry	T-24	BGC-823	L-02	3a	5.6	34	147
3b	12	18	98	3a	5.6	34	147
3b	12	18	98	3c	8.7	37	86

As known from Table 3, compound 3a～c has good antineoplastic activity, and less to the toxic action of normal liver epithelial cell L-02.Further activity research well afoot.

The thiophosphatephosphorothioate compounds that contains the Tegafur of selenium is that a class is not seen the bibliographical information new compound.It is synthetic to be by " two steps treated different things alike ", is solvent with the dry-out benzene, under 65 ℃, makes 2-N with the iodine powder as catalyzer, N-diethylamino-1,3,2-dioxaphospholane successively with hydroxyalkyl Tegafur and sulphur powder reaction synthetic intermediate (2); Then, in room temperature, dehydrated alcohol, the fragrant sodium selenide that sodium borohydride reduction diaryl diselenide produces carries out nucleophilic ring opening to intermediate (2), and the formation of high yield contains the thiophosphatephosphorothioate compounds of the Tegafur of selenium.Their structure is passed through ³¹P NMR, ¹H NMR and mass spectrum confirmation.

Embodiment

1, the group thiophosphate compound that contains the Tegafur of selenium synthesizes

1.1 contain 1,3 of Tegafur, 2-dioxaphospholane (2) synthetic:

In the 25ml dry-out benzene, add 3-hydroxyethyl Tegafur 732mg (3.0mmol), 2-N successively; N-diethylamino-1; 3; 2-dioxaphospholane 978mg (6.0mmol) and 76mg (0.3mmol) iodine powder, nitrogen protection, 65 ℃ of stirrings of oil bath (800 rev/mins) 24 hours; adding 192mg (6.0mmo1) sulphur powder then continues to stir 0.5 hour; solvent is removed in rotation, residue column chromatography (ethyl acetate/sherwood oil=1/1) separate compound (2) 638mg, colourless viscous liquid.

2.2 containing the group thiophosphate compound of the Tegafur of selenium synthesizes

In the there-necked flask of 50ml, add 76mg (2.0mmol) sodium borohydride, 10ml dehydrated alcohol, 312mg (1.0mmol) diselenide successively; about 50 minutes of stirring at room; become clarification up to reaction system; extract dehydrated alcohol (2ml) solution that the selenophenol sodium solution 5ml for preparing injects 366mg (1.0mmol) compound (2) with syringe; nitrogen protection; stirred overnight at room temperature; uncovered continuation was stirred 12 hours; column chromatography (methyl alcohol/trichloromethane=1/5) got compound (3a) 481mg, white powder after solvent was removed in decompression.

2, antitumor cytolytic activity

With RPIM 1640 substratum that contain 10% newborn calf serum (training base) fully with cell cultures to be measured to logarithmic phase, be digested to individual cells with the Digestive system that contains 0.25% pancreatin, 0.02%EDTA, counting, inoculating cell is in 96 well culture plates, and 5000 cells in every hole comprise 5 dosage groups, every group of 6 parallel holes, at 37 ℃, 100% relative humidity contains 5%CO ₂, 95% air incubator in pre-the cultivation 24 hours.

Difference assay takes by weighing testing sample 30mg, and with the 3mlDMSO dissolving, adding secondary deionized water to cumulative volume is 30ml, and promptly mother liquid concentration is 1000ug/ml.

The substratum that contains the different concns testing sample that test group renews: 10ug/ml: get mother liquor 0.12ml and 11.82ml and train base fully and be added to 6 and cultivate in the plate holes; 20ug/ml: get mother liquor 0.24ml and 11.76ml and train base fully and be added to 6 and cultivate in the plate holes; 50ug/ml, 100ug/ml, 200ug/ml prepares with similar method.Control group then changes 12ml into and trains base fully.

Cell is at 37 ℃, and 100% relative humidity contains 5%CO ₂, 95% air incubator in cultured continuously after 24 hours, every hole adds the serum-free medium that contains 0.2mg/mlMTT of the new preparation of 20ul, continues to cultivate 4 hours.

Inhale carefully and remove supernatant liquid, every hole adds 100ulDMSO, and vibration makes the crystallisate of metabolism generation dissolve the back fully and measures the OD value at 570nm wavelength place with microplate reader gently.

Thank you: this paper work obtains state natural sciences fund (20372020) and subsidizes.

Claims

1. thiophosphatephosphorothioate compounds that contains the Tegafur of selenium is characterized in that the general formula of this compound is:

Ar＝C ₆H ₅，p-CH ₃C ₆H ₄，m-CH ₃C ₆H ₄，

2. the thiophosphatephosphorothioate compounds of a Tegafur that contains selenium as claimed in claim 1 is synthetic.Method is as follows: be solvent with the dry-out benzene, under 65 ℃, make 2-N as catalyzer with the iodine powder, N-diethylamino-1,3,2-dioxaphospholane successively with hydroxyalkyl Tegafur and sulphur powder reaction synthetic intermediate (2); Then, in room temperature, dehydrated alcohol, the fragrant sodium selenide that sodium borohydride reduction diaryl diselenide produces carries out nucleophilic ring opening to intermediate (2), and the formation of high yield contains the thiophosphatephosphorothioate compounds of the Tegafur of selenium, and reaction formula is:

Ar＝C ₆H ₅，p-CH ₃C ₆H ₄，m-CH ₃C ₆H ₄