CN103980174A - Substituted bismuth pyrrolidinyl dithiocarbamate (III) complex and preparation method thereof, and application of complex in preparing antineoplastic drugs - Google Patents
Substituted bismuth pyrrolidinyl dithiocarbamate (III) complex and preparation method thereof, and application of complex in preparing antineoplastic drugs Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/06—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with radicals, containing only hydrogen and carbon atoms, attached to ring carbon atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
Abstract
The invention relates to a substituted bismuth pyrrolidinyl dithiocarbamate (III) complex of which the structural formula is disclosed in the specification. The substituted bismuth pyrrolidinyl dithiocarbamate (III) complex has favorable antitumor activity, has obvious activity for inhibiting stomach cancer, lung cancer, liver cancer, colorectal cancer, cervical carcinoma, oophoroma, mammary cancer, leukaemia, colon cancer and other malignant tumors, and has wide application prospects when being developed into novel antitumor drugs.
Description
technical field
The present invention relates to antitumor drug technical field, be specifically related to substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex and preparation method thereof and in the application of preparing in antitumor drug.
Background technology
The mankind's health and lives in malignant tumour (cancer) serious threat, the patient who dies from every year cancer accounts for 1/4th of total death toll, and sickness rate is increase year after year situation, the control of cancer becomes one of problem that medical science and life science worker mainly study.China's cancer morbidity is very surprising, has become second-biggest-in-the-world cancer state occurred frequently, and statistic data (WHOGLOBOCAN 2008) shows: all tumour new cases of China are 2,800,000, are 2 times (U.S. are 1,400,000) of the new tumor cases of the U.S.; The same year, China's tumor mortality case was 1,960,000, and the death of the U.S. is 570,000, and the gap of the two is 3.4 times.As can be seen here, the having a high potential of Science in Future in China and global cancer therapy drug market growth, research and development new generation anti-cancer medicament must have huge market.But, have tens kinds for the cancer therapy drug of cancer therapy at present, but the common solid tumor of majority is still lacked to active drug, and many antitumour drugs produce resistance in various degree in process of clinical application.Therefore, the antitumor drug of exploitation screening high-efficiency low-toxicity becomes anti-cancer agent and researches and develops of paramount importance task.
Bismuth has long medicinal history.Bismuth belongs to period 6 V A family heavy metal element in periodictable, is positioned at metal and nonmetal intersection, has special physico-chemical property.Due to nontoxicity, the non-carinogenicity of bismuth, be called as green metal.Through years of researches and development, bismuth compound has been widely used in the aspects such as medical science, chemical industry and biology, the field being wherein most widely used is medicine, as for Surgery Treatment wound and hemostasis, as the pharmaceutical cpd of killing Helicobacter pylori treatment stomach ulcer and gastrointestinal disturbance, separately studies have found that the growth that the title complex of bismuth can anticancer and have no side effect.
To bismuth preparation, the effect in medical treatment conducts in-depth research professor Sun Hongzhe of Hong Kong University.Bismuth compound not only shows to have on antitumour activity in the application of field of cancer, and they can also alleviate the caused toxic side effect of other anticancer chemotherapeutic agent.Experimentation on animals shows, Bismuth trinitrate and citric acid coupling can be alleviated effectively along platinum medicine caused kidney damage in anticancer therapy process.Research discovery recently, bismuth agent can suppress the growth of sars coronavirus, and the helicase (helicase) in its process same Bi of possibility (III) inhibition SARS virus is relevant.Bismuth can with organism in different kinds of molecules combination.Research finds that gsh (GSH) can prevent CBS precipitation.Bismuth also can by and Transferrins,iron complexes (transferrin) in conjunction with the object that reaches biological transmission.
Within 2006, the Edward R T Tiekink of NUS has taught at U. S. application the patent of Bismuth dibutyldithiocarbamate compound (US 2006/0142621 A1), has inquired into the anti-tumor activity of dialkyl amino dithioacid bismuth compound.But exist structure single, the antitumor cell strain of effect is few, the problem that clinical value is lower, also has larger distance apart from clinical application.
Summary of the invention
The object of the invention is to open a kind of substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex and preparation method thereof and in the application of preparing in antitumor drug, substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex has good anti-tumor activity, and the malignant tumours such as cancer of the stomach, lung cancer, liver cancer, large bowel cancer, cervical cancer, ovarian cancer, mammary cancer, leukemia, colorectal carcinoma are had to significant antitumor activity.
For achieving the above object, the technical scheme adopting is in the present invention: substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex, and skeleton symbol is as follows:
,
R
2and R
3for H, R
1for
;
or
;
Or, R
1and R
3for H, R
2for F;
Or, R
1and R
2for H, R
3for F;
Or, R
1for H, R
2and R
3for F.
The preparation method of substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex:
Get (2R)-2-crassitude and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids recrystallization, obtains (2R)-2-crassitude dithiocarbamic acid bismuth;
Or: get L-PROLINE and triethylamine adds anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, reaction finishes rear adjust pH, washes out after precipitation and carries out suction filtration, alcohol wash, dry, and solids recrystallization, obtains (S)-2-carboxy pyrrole alkane dithiocarbamic acid bismuth;
Or: get L-PROLINE carbethoxy hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, after reaction finishes, carry out suction filtration, alcohol wash, dry, solids recrystallization, (S)-2-ethoxycarbonyl tetramethyleneimine dithiocarbamic acid bismuth;
Or: get R-3-fluoropyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains (3R)-3-fluoropyrrolidine dithiocarbamic acid bismuth;
Or: get S-3-fluoropyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains (3S)-3-fluoropyrrolidine dithiocarbamic acid bismuth;
Or: get 3,3-difluoro pyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains 3,3-, bis-fluoropyrrolidine dithiocarbamic acid bismuths;
Substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex has significant antitumor activity to malignant tumours such as cancer of the stomach, lung cancer, liver cancer, large bowel cancer, cervical cancer, ovarian cancer, mammary cancer, leukemia, colorectal carcinomas.
beneficial effect
Salt and the hydrate of substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex of the present invention and described title complex have good anti-tumor activity, preparation method is simple, the malignant tumours such as cancer of the stomach, lung cancer, liver cancer, large bowel cancer, cervical cancer, ovarian cancer, mammary cancer, leukemia, colorectal carcinoma are had to significant antitumor activity, be developed as new antitumoral new drug and be with a wide range of applications.
Brief description of the drawings
Fig. 1-Fig. 4 is that the title complex (numbering N048) of embodiment tri-suppresses test result for tumor cell extracorporeal growth;
Fig. 5-Fig. 7 is that the title complex (numbering N055) of embodiment tetra-suppresses test result for tumor cell extracorporeal growth;
Fig. 8-Figure 10 is that the title complex (numbering N060) of embodiment five suppresses test result for tumor cell extracorporeal growth;
Figure 11-Figure 13 is that the title complex (numbering N061) of embodiment six suppresses test result for tumor cell extracorporeal growth;
Figure 14-Figure 16 is that the title complex (numbering N062) of embodiment mono-suppresses test result for tumor cell extracorporeal growth;
Figure 17 is the title complex (numbering N048) of the embodiment tri-growth inhibition test result to noumenal tumour model;
Figure 18 is the title complex (numbering N060) of the embodiment five growth inhibition test result to noumenal tumour model.
Embodiment
By specific embodiment, the present invention is further illustrated below, but not as restriction.
embodiment mono-:(2R)-2-crassitude dithiocarbamic acid bismuth and preparation thereof
In round-bottomed flask, add 0.426 g (5 mmol) (2R)-2-crassitude, 0.25g (6mmol) sodium hydroxide, 15mL anhydrous methanol, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76g (10mmol) CS
2, magnetic agitation is reacted 5 h.
By 0.53g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 3 h, suction filtration, anhydrous methanol is washed, vacuum-drying.Methylene dichloride and ethyl alcohol recrystallization for solids, obtain 0.82 g yellow powder solid, yield 71.3 %.Fusing point: 224-226 DEG C, specific rotatory power
=-205
°(HCl
3), be numbered N062.
1HNMR(400?MHz,?CDCl
3),?δ:?1.362～1.378?(9H,?d,?-CH
3,?J=6.4Hz),?1.692～1.731?(3H,?m,?Cy-CH，J=5.2Hz),?2.035～2.071(3H,?m,?Cy-CH,?J=4.0-4.8Hz),?2.088～2.161(6H,?m,?Cy-CH
2-,?J=4.0-6.8Hz),?3.890～3.952?(6H,?q,?N-CH
2,?J=7.6-9.2Hz),?4.725～4.782?(3H,?m,?N-CH,?J=6.4-6.8Hz)。
Molecular formula: C
18h
30n
3s
6bi, results of elemental analyses (being calculated value % in bracket): C 31.29 (31.34), H 4.30 (4.39), N 6.06 (6.09).
embodiment bis-:(S)-2-carboxy pyrrole alkane dithiocarbamic acid bismuth and preparation thereof
In 50 mL round-bottomed flasks, add 0.576 g (5mmol) L-PROLINE, 0.759 g (7.5mmol) triethylamine, 15mL anhydrous methanol, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76g (10mmol) CS
2, magnetic agitation is reacted 1 h, and stirring at room temperature is reacted 3 h.
By 0.53 g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 4 h.With glacial acetic acid adjust pH be 5-6, separate out yellow mercury oxide, suction filtration, anhydrous methanol is washed, dry.Solids recrystallizing methanol, obtains 1.05 g yellow solids.Yield 80.8%.Fusing point: 186-188 DEG C, specific rotatory power
=-103
°(DMSO).
1HNMR(400?MHz,?DMSO-
d 6),?δ:?1.971～2.014?(6H,?m,?Cy-CH
2,?J=6.4～7.2Hz),?2.272～2.318?(6H,?m,?Cy-CH
2，J=6.4-7.6Hz),?3.818～3.848(6H,?t,?N-CH
2,?J=7.6Hz),?4.761～4.805?(3H,?t,?N-CH,?J=8.8Hz)。
Molecular formula: C
18h
24n
3o
6s
6bi, results of elemental analyses (being calculated value % in bracket): C 27.69 (27.72), H 3.01 (3.11), N 5.32 (5.39).
embodiment tri-:(S)-2-ethoxycarbonyl tetramethyleneimine dithiocarbamic acid bismuth and preparation thereof
In 50 mL round-bottomed flasks, add 0.898g (5 mmol) L-PROLINE carbethoxy hydrochloride, 0.44 g (11 mmol) sodium hydroxide, 15 mL anhydrous methanols, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76 g (10 mmol) CS
2, magnetic agitation is reacted 2 h, and stirring at room temperature is reacted 4 h.
By 0.53 g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 3 h, suction filtration, anhydrous methanol is washed, vacuum-drying.Solids acetonitrile recrystallization, obtains 1.15 g aureus crystal.Yield 79.9%.Fusing point: 89-91 DEG C, specific rotatory power
=-393
°(CHCl
3), be numbered N048.
1HNMR(400?MHz,?CDCl
3),?δ:?1.261～1.297?(9H,?t,?-CH
3,?J=7.2Hz),?2.069～2.091?(3H,?m,?Cy-CH，J=4.4-6.8Hz),?2.092～2.155?(6H,?m,?Cy-CH
2,?J=5.6-8.2Hz),?2.314～2.400?(3H,?m,?Cy-CH，J=6.4-8.0Hz),?3.929～3.998(3H,?m,?N-CH,?J=4.4-7.6Hz),?4.061～4.108?(3H,?m,?N-CH,?J=3.2-5.2Hz),?4.185～4.238?(2H,?q,?O-CH
2,?J=7.2Hz),?4.997～5.031?(3H,?d?d,?N-CH,?J=3.2-7.6Hz)。
Molecular formula: C
24h
36n
3o
6s
6bi, results of elemental analyses (being calculated value % in bracket): C 33.28 (33.36), H 4.16 (4.21), N 4.69 (4.86).
embodiment tetra-:(3R)-3-fluoropyrrolidine dithiocarbamic acid bismuth and preparation thereof
In round-bottomed flask, add 0.628 g (5mmol) R-3-fluoropyrrolidine hydrochloride, 0.44 g (11mmol) sodium hydroxide, 15 mL anhydrous methanols, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76 g (10 mmol) CS
2, magnetic agitation is reacted 2 h, and stirring at room temperature is reacted 4 h.
By 0.53 g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 3 h, suction filtration, anhydrous methanol is washed, dry.Solids is purifying on post, eluent: ethyl acetate-sherwood oil=1:2, separates and obtain 1.02 g yellow powder solids.Yield 87.2 %, fusing point: 156-158 DEG C, specific rotatory power
=+10.5
°(CHCl
3), be numbered N048.
1HNMR(400?MHz,?CDCl
3),?δ:?2.349～2.442?(6H,?m,?Cy-CH
2,?J=6.0-8.0Hz),?3.883～3.960?(6H,?m,?N-CH
2,?J=7.4Hz),?4.285～4.315?(3H,?m,?F-CH，J=8.0Hz),?5.255～5.286(3H,?q,?N-CH,?J=5.6～7.6Hz)，5.388～5.416?(3H,?q,?N-CH,?J=4.8～?6.8Hz)。
Molecular formula: C
15h
21n
3f
3s
6bi, results of elemental analyses (being calculated value % in bracket): C 25.68 (25.67), H 3.01 (3.02), N 5.89 (5.99).
embodiment five:(3S)-3-fluoropyrrolidine dithiocarbamic acid bismuth and preparation thereof
In round-bottomed flask, add 0.628 g (5mmol) S-3-fluoropyrrolidine hydrochloride, 0.44 g (11mmol) sodium hydroxide, 15 mL anhydrous methanols, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76 g (10 mmol) CS
2, magnetic agitation is reacted 2 h, and stirring at room temperature is reacted 4 h.
By 0.53 g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 3h, suction filtration, anhydrous methanol is washed, dry.Solids is purifying on post, eluent: ethyl acetate-sherwood oil=1:2, separates and obtain 0.97 g yellow powder solid.Yield 82.9%, fusing point: 156-158 DEG C, specific rotatory power
=-10.5
°(CHCl
3), be numbered N060.
1HNMR(400?MHz,?CDCl
3),?δ:?2.349～2.442?(6H,?m,?Cy-CH
2,?J=6.0-8.0Hz),?3.883～3.960?(6H,?m,?N-CH
2,?J=7.4Hz),?4.285～4.315?(3H,?m,?F-CH，J=8.0Hz),?5.255～5.286(3H,?q,?N-CH,?J=5.6～7.6Hz)，5.388～5.416?(3H,?q,?N-CH,?J=4.8～?6.8Hz)。
Molecular formula: C
15h
21n
3f
3s
6bi, results of elemental analyses (being calculated value % in bracket): C 25.63 (25.67), H 2.98 (3.02), N 5.88 (5.99).
embodiment six:3,3-, bis-fluoropyrrolidine dithiocarbamic acid bismuth and preparations thereof
In round-bottomed flask, add 0.718 g (5mmol) 3,3-difluoro pyrrolidine hydrochloride, 0.44 g (11 mmol) sodium hydroxide, 15 mL anhydrous methanols, after magnetic agitation is dissolved, 0-5 DEG C of cooling bath, drips 0.76g (10 mmol) CS
2, magnetic agitation is reacted 2 h, and stirring at room temperature is reacted 4 h.
By 0.53 g (1.7mmol) BiCl
3be dissolved in 15 mL anhydrous methanols, splash in above-mentioned reaction solution, stirring at room temperature 5 h, suction filtration, anhydrous methanol is washed, dry.Solids is purifying on post, eluent: ethyl acetate-sherwood oil=1:3, separates and obtain 1.11 g yellow powder solids.Yield 88.1%, fusing point: 228-230 DEG C, is numbered N061.
1HNMR(CDCl
3,?400?MHz),?δ:?2.521～2.557?(6H,?t,?Cy-CH
2,?J=7.2Hz),?4.220～4.262?(6H,?t,?N-CH
2,?J=8.4Hz),?5.405?(6H,?s,?N-CH
2)。
Molecular formula: C
15h
18n
3f
6s
6bi, results of elemental analyses (being calculated value % in bracket): C 23.73 (23.84), H 2.38 (2.41), N 5.48 (5.56).
antitumor cytolytic activity
one, anti tumor activity in vitro research
1. cell strain and cultivation
The cancer cell such as cancer cell leukemia cell line (HL-60), lung cancer cell line (A-549), hepatoma cell strain (BEL-7402), colorectal cancer cell lines (SW-1116), cervical cancer cell strain (HELA), Ovarian Cancer Cells (3AO), breast carcinoma cell strain (MCF-7), stomach cancer cell line (MKN-28) are incubated to 10% inactivated fetal bovine serum, 100U/mL penicillin, in the RPMI1640 or DMEM substratum of 100 μ g/mL Streptomycin sulphates, in 10% CO of 37 DEG C
2under incubator and saturated humidity condition, cultivate.For normal cultivation, attached cell is all with carrying out in the trypsinase-EDTA of 1 times of concentration after the de-wall of tryptic digestion, by the ratio of 1:5～1:20 by within every cell 3～4 days, going down to posterity once (when cell is when in culture dish, institute's area coverage is 80-90%).
2. Antitumor Activity of Drugs is measured
Test the day before yesterday, described cell carries out tryptic digestion approximately 5 minutes (in 37 DEG C of incubators) in 1 times of concentration trypsinase-EDTA solution, is suspended in the DMEM or RPMI nutrient solution of 10 mL, is then inoculated into 96 orifice plates.Cell is 1～2 × 10 by density
4/ hole is inoculated in standard DMEM or RPMI substratum, and volume is 50 μ L.
Second day, prepare a series of 2x concentration dilutions of tested metal complexes with standard DMEM nutrient solution and store liquid (0.003 μ M～60 μ M).50 μ L dilution storage liquid are joined in corresponding plate well, and the tested title complex ultimate density that the cell in test slab is accepted is (0.0015 μ M～30 μ M) a series of 9 different dosage groups, every group of 3 parallel holes.Therefore 2 kinds of compounds (for example X and Y) can be tested (table 1) in a plate simultaneously.Except test compound, each plate also comprises positive control (0.5 μ M Zorubicin) and negative control (DMSO).
Table 1: the testing scheme of 96 orifice plates and tested compounds final concentration in test
96 orifice plate culture plates are put in 37 DEG C of incubators and are cultivated after 48 hours, and abandoning supernatant adds the serum-free medium of the 0.5 mg/mL MTT solution (0.5%MTT) that contains 20 μ L of the 200 fresh configurations in μ L/ hole, and 37 DEG C are continued to cultivate 5 hours.Abandoning supernatant, add 150 μ L DMSO, after concussion mixes, measure the light absorption value in each hole at enzyme-linked immunosorbent assay instrument OD 490 nm places, calculate inhibitory rate of cell growth, formula is: inhibitory rate of cell growth=[(negative control group OD value-experimental group OD value)/negative control group OD value] × 100%.And by inhibitory rate of cell growth and corresponding concentration, go out IC50 value (half inhibiting rate IC50, the drug level that inhibitory rate of cell growth is 50%) by computed in software.For each compound, this mensuration all repeats twice in addition, once complete mensuration three times, just calculates IC50 value by raw data.
3. experimental result
Measure numbering N048, N055, N060, N061, the N062 etc. of different concns through mtt assay for tumor cell growth activity impact, result is as shown in Fig. 1-Figure 16.Numbering N048, N055, N060, N061, N062 etc. suppress active IC50 value (table 2) as shown in the table to various tumor cell extracorporeal growths.
Table 2. title complex suppresses active to tumor cell extracorporeal growth
two, anti-tumor in vivo activity research
1. experiment material
1.1. laboratory animal and knurl strain
8-12 health Balb/c in age in week mouse, male and female half and half (body weight 22-25g), each cage fills 5 mouse, is placed between clean animal raising the drinking-water of freely ingesting.
Use mouse colonic cell C26 is the subcutaneous allos noumenal tumour model of making in Balb/c mouse both sides.
1.2. testing drug
All title complexs are all formulated in 20%DMSO carrier soln, and to numbering N048, concentration is 4 mg/mL; To numbering N060, concentration is 4 mg/mL.
2. experimental procedure
2.1. animal model and grouping
Before mouse both sides, tuck in lower to subcutaneous injection 5 × 10
6individual culturing cell has been set up C26 solid tumor models, treats that 80% gross tumor volume is greater than 80 mm
3time, all mouse are divided into embodiment treatment group and vehicle Control group at random.
2.2. medication
All described title complexs are all with intraperitoneal injection, 30mg/kg/ days, control group is accepted, without drug carrier injection, within one week 5 days, to continue 3 weeks, have a rest one week, or until mouse have to put to death due to tumour greatly.
2.3. measurement of tumor
Two axles (mm) (L, the major axis of vernier caliper measurement tumour; W, minor axis).With formula: gross tumor volume V=0.5 × L × W
2estimate gross tumor volume V(cubic millimeter, mm
3).In pharmacological agent, carry out measurement of tumor every day, after treatment finishes one week 3-4 time.
2.4. the evaluation of antitumous effect
Anti-tumor activity is by drawing tumor growth curve and calculating maximum inhibition rate of tumor growth (maximum tumour inhibiting rate, MTGI) and evaluate: maximum tumor growth suppresses (MTGI)=[(vehicle Control group mean tumour volume-administration group mean tumour volume)/vehicle Control group mean tumour volume] × 100%.
2.5. experimental result
Result statistical analysis, the maximum tumour inhibiting rate of numbering N048 is 16%, p<0.05; The maximum tumour inhibiting rate of numbering N060 is 10%, p<0.05.The tumor growth curve of drawing after one-way ANOVA analyzes is as Figure 17 and Figure 18.
3. conclusion
Experimental data shows, metal complexes has significant antitumor activity to malignant tumours such as cancer of the stomach, lung cancer, liver cancer, large bowel cancer, cervical cancer, ovarian cancer, mammary cancer, leukemia, colorectal carcinomas, is developed as new antitumoral new drug and is with a wide range of applications.
Claims (4)
1. substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex, is characterized in that: the skeleton symbol of described substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex is as follows:
,
R
2and R
3for H, R
1for
;
or
;
Or, R
1and R
3for H, R
2for F;
Or, R
1and R
2for H, R
3for F;
Or, R
1for H, R
2and R
3for F.
2. the preparation method of substituted pyrrolidin dithiocarbamic acid bismuth as claimed in claim 1 (III) title complex, is characterized in that:
Get (2R)-2-crassitude and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids recrystallization, obtains (2R)-2-crassitude dithiocarbamic acid bismuth;
Or: get L-PROLINE and triethylamine adds anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, reaction finishes rear adjust pH, separates out after precipitation and carries out suction filtration, alcohol wash, dry, and solids recrystallization, obtains (S)-2-carboxy pyrrole alkane dithiocarbamic acid bismuth;
Or: get L-PROLINE carbethoxy hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, after reaction finishes, carry out suction filtration, alcohol wash, dry, solids recrystallization, (S)-2-ethoxycarbonyl tetramethyleneimine dithiocarbamic acid bismuth;
Or: get R-3-fluoropyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains (3R)-3-fluoropyrrolidine dithiocarbamic acid bismuth;
Or: get S-3-fluoropyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains (3S)-3-fluoropyrrolidine dithiocarbamic acid bismuth;
Or: get 3,3-difluoro pyrrolidine hydrochloride and NaOH and add anhydrous methanol, after stirring and dissolving, drip CS
2, obtain reaction solution, get BiCl
3splash in above-mentioned reaction solution with the mixing solutions of anhydrous methanol, carry out suction filtration, alcohol wash, dry after reaction finishes, solids purifies and separates on silicagel column obtains 3,3-, bis-fluoropyrrolidine dithiocarbamic acid bismuths.
3. the salt of substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex as claimed in claim 1 and described title complex and hydrate are in the application of preparing in antitumor drug.
4. the salt of substituted pyrrolidin dithiocarbamic acid bismuth (III) title complex as claimed in claim 3 and described title complex and hydrate, in the application of preparing in antitumor drug, is characterized in that: described tumour is cancer of the stomach, lung cancer, liver cancer, large bowel cancer, cervical cancer, ovarian cancer, mammary cancer, leukemia or colorectal carcinoma.
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| PCT/CN2015/080495 WO2015180693A1 (en) | 2014-05-30 | 2015-06-01 | Bismuth(iii)dithiocarbamate complex and preparation method and use thereof |
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| CN104876893A (en) * | 2015-05-25 | 2015-09-02 | 南阳师范学院 | Substituted-homopiperzine dithiocarbamic acid bismuth complexes, and medicinal salts, preparation methods and antitumor application thereof |
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|---|---|---|---|---|
| WO2004067505A2 (en) * | 2003-01-29 | 2004-08-12 | National University Of Singapore | Bismuth dithiocarbamate compounds and uses thereof |
| US20100137557A1 (en) * | 2005-07-07 | 2010-06-03 | Cancure Laboratories Llc | Use of one or more metal carriers to selectively kill mammalian cells |
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| WO2004067505A2 (en) * | 2003-01-29 | 2004-08-12 | National University Of Singapore | Bismuth dithiocarbamate compounds and uses thereof |
| US20100137557A1 (en) * | 2005-07-07 | 2010-06-03 | Cancure Laboratories Llc | Use of one or more metal carriers to selectively kill mammalian cells |
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| CN104876893A (en) * | 2015-05-25 | 2015-09-02 | 南阳师范学院 | Substituted-homopiperzine dithiocarbamic acid bismuth complexes, and medicinal salts, preparation methods and antitumor application thereof |
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