Ruthenium complex containing alkynyl group, method of synthesizing the same and use thereof
11420990 · 2022-08-23
Assignee
Inventors
Cpc classification
C09K2211/185
CHEMISTRY; METALLURGY
C09K2211/1074
CHEMISTRY; METALLURGY
B01J31/2295
PERFORMING OPERATIONS; TRANSPORTING
International classification
C07F15/00
CHEMISTRY; METALLURGY
Abstract
A Ruthenium complex containing alkynyl group, a method of synthesizing the Ruthenium complex containing alkynyl group and a use thereof are provided. The Ruthenium complex has a chemical formula of Ru(L).sub.2(DPPZ′). DPPZ′ is a main ligand having structural formula of ##STR00001##
R is any one selected from the group consisting of H, substituted or unsubstituted phenyl, R.sub.1NH.sub.2, R.sub.1OH, and SiMe.sub.3; R.sub.1 is C1-C5 chain alkyl group; L is an auxiliary ligand with N as coordinating atom. Sonogashira coupling reaction is utilized to introduce alkynyl group into a DPPZ-type Ruthenium complex; the introduced alkynyl group is beneficial to promote the transmembrane absorption of drug molecules, increase the probability of drug entry into cells, and can also increase efficacy and reduce toxic and side effects of drugs. The Ruthenium complex provided has significant anti-tumor activity, especially anti-breast cancer activity, and can provide new ideas for designing anti-tumor drug molecules in the future.
Claims
1. A method of synthesizing a Ruthenium complex containing an alkynyl group, comprising the following steps: S1: adding Br-DPPZ, [Ru(L).sub.2Cl.sub.2].nH.sub.2O or L-[Ru(L).sub.2(py).sub.2].mH.sub.2O or D-[Ru(L).sub.2(py).sub.2].mH.sub.2O and a mixed solvent to a reaction vessel to obtain a first mixed solution; afterwards, heating and refluxing the first mixed solution for a first reaction to obtain a first resulting product, when the first reaction is complete, post-processing the first resulting product to obtain an intermediate; wherein py represents pyridine, and each of n and m is an integer, independently; S2: adding the intermediate prepared in S1 into a microwave reaction tube, and adding anhydrous acetonitrile to the intermediate to obtain a mixture, and performing an ultrasonic treatment on the mixture to dissolve the intermediate to obtain a second mixed solution; afterwards, under a protection of an inert gas, first adding triethylamine and ##STR00044## and then immediately adding Pd(PPh.sub.3).sub.2Cl.sub.2 and CuI to the second mixed solution; after sealing the microwave reaction tube, putting the microwave reaction tube into a microwave reactor for a microwave assisted heating for a second reaction to obtain a second resulting product, and when the second reaction is complete, post-processing the second resulting product to obtain the Ruthenium complex containing the alkynyl group, wherein a chemical formula of the Ruthenium complex containing the alkynyl group is Ru(L).sub.2(DPPZ′), wherein L is an auxiliary ligand with N as coordinating atom, and DPPZ′ is a main ligand having structural formula (I) as follows: ##STR00045## wherein R is any one selected from the group consisting of H, substituted phenyl, unsubstituted phenyl, R.sub.1NH.sub.2, R.sub.1OH, and SiMe.sub.3, wherein R.sub.1 is a C1-C5 chain alkyl group.
2. The method according to claim 1, wherein the mixed solvent is ethylene glycol and water in S1.
3. The method according to claim 2, wherein a volume ratio of the ethylene glycol and the water is 9:1 in S1.
4. The method according to claim 1, wherein the inert gas is nitrogen or argon in S2.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
(1) The present invention will be further described below with reference to the accompanying drawings:
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DETAILED DESCRIPTION OF THE EMBODIMENTS
(20) 1. The Molecular Structure and Number of the Designed Compound
(21) 1.1. The Molecular Structure and Number of the Ligand
(22) ##STR00010##
(23) 1.2. The Molecular Structure and Number of DPPZ Alkynyl Ruthenium Complex
(24) 1.2.1. The Molecular Structure and Number of Racemic Bipyridine DPPZ Alkynyl Ruthenium Complex
(25) ##STR00011## ##STR00012##
(26) 1.2.2. The Molecular Structure and Number of Chiral Bipyridine DPPZ Alkynyl Ruthenium Complex
(27) ##STR00013## ##STR00014## ##STR00015## ##STR00016##
(28) 1.2.3. The Molecular Structure and Number of Racemic Phenanthroline DPPZ Alkynyl Ruthenium Complex
(29) ##STR00017## ##STR00018##
(30) 2. The Overall Synthesis Route of Alkynyl Modified Ruthenium(II) Complex with DPPZ as the Main Ligand
(31) 2.1. The Microwave Assisted Synthesis Route of Racemic Bipyridine DPPZ Alkynyl Complex
(32) ##STR00019##
(33) 2.2 The Microwave Assisted Synthesis Route of Chiral Bipyridine DPPZ Alkynyl Complex
(34) ##STR00020## ##STR00021##
(35) Description of Chiral Resolution
(36) 1) Synthesis of [Ru(bpy).sub.2Cl.sub.2].2H.sub.2O
(37) Bipyridine (1.87 g, 12 mmol), lithium chloride (2.43 g, 57.6 mmol), and ruthenium trichloride (1.57 g, 6 mmol) were added into a 50 mL three-necked flask, then DMF and water were added, the reaction was performed by refluxing at 140° C. for 8 h under the protection of argon. After the reaction stopped, the reaction system was cooled to room temperature, then acetone was added, and black crystals were obtained by vacuum filtration; after washing a filter cake for several times with ice water and acetone, it was dried in a vacuum drier to obtain purple-black crystals with a yield of 75.5%. (Calculated with RuCl.sub.3.3H.sub.2O).
(38) 2) Synthesis of [Ru(bpy).sub.2(py).sub.2].2H.sub.2O
(39) [Ru(bpy).sub.2Cl.sub.2].2H.sub.2O (2.01 g, 3.85 mmol), pyridine and distilled water were added into a 150 mL three-necked flask, the reaction was performed by refluxing at 100° C. for 4 h under the protection of argon. After the reaction stopped, the reaction system was cooled to room temperature, all the solvent was evaporated under reduced pressure; afterwards, the resulting residue was dissolved in methanol to obtain a red solution, then ether was added, before performing vacuum filtration; after washing a filter cake for several times with ether, it was dried in a vacuum drier to obtain red crystals with a yield of 89.5%.
(40) 3) Synthesis of [Ru(bpy).sub.2(py).sub.2][o,o′-dibenzoyl-D-tartrate].12H.sub.2O
(41) 1.95 g Cis-[Ru(bpy).sub.2(py).sub.2].Cl.sub.2 was dissolved in water, 0.5 M O,O′-dibenzoyl D-sodium tartrate solution was added at room temperature (O,O′-dibenzoyl D-sodium tartrate solution was prepared by neutralizing O,O′-dibenzoyl D-tartaric acid with sodium hydroxide), stirring at room temperature for 25 min; after filtration, a filtrate was placed in a beaker for crystallizing in a fume hood. After ten days, a large amount of red crystals were precipitated. The crystals were washed for several times with ether and dried in a vacuum dryer to obtain red acicular crystals, with a yield of 45.8%.
(42) 4) Synthesis of [Ru(bpy).sub.2(py).sub.2][o,o′-dibenzoyl-L-tartrate].12H.sub.2O
(43) 1.95 g Cis-[Ru(bpy).sub.2(py).sub.2].Cl.sub.2 was dissolved in water, 0.5 M O,O′-dibenzoyl L-sodium tartrate solution was added at room temperature (O,O′-dibenzoyl L-sodium tartrate solution was prepared by neutralizing O,O′-dibenzoyl L-tartaric acid with sodium hydroxide), stirring at room temperature for 25 min; after filtration, a filtrate was placed in a beaker for crystallizing in a fume hood. After ten days, a large amount of red crystals were precipitated. The crystals were washed for several times with ether and dried in a vacuum dryer to obtain red acicular crystals, with a yield of 46.2%.
(44) 2.3. The Microwave Assisted Synthesis Route of Racemic Phenanthroline DPPZ Alkynyl Complex
(45) ##STR00022##
Example 1. Synthesis of a Ligand Br-DPPZ
(46) ##STR00023##
(47) 315.16 mg (1.5 mmol) o-phenanthroline 5,6-dione and 232.48 mg (1.25 mmol) 4-bromo-o-phenylenediamine were weighed and taken into a 30 ml quartz tube, and 20 ml absolute ethanol was added thereto, stirring for 5 minutes, and performing microwave reaction at 95° C. for 3 minutes. After the completion of the reaction, the reaction system was cooled to room temperature, 30 ml water was added, and a yellow-white solid was precipitated; after vacuum filtration, a filter cake was washed with water and dried in a vacuum drying oven at 50° C. The obtained crude product was dissolved in pure chloroform, packed in column with pure chloroform, and was purified by eluting with an eluent (pure chloroform:absolute ethanol=100:1). Afterwards, a main yellow band was collected and was subjected to rotary evaporation under reduced pressure to obtain a yellow solid, which was recrystallized with a solvent of absolute ethanol:chloroform=3:2, and a yield was 80%.
Example 2. Synthesis of [Ru(bpy).SUB.2.(Br-DPPZ)].SUP.2+
(48) ##STR00024##
(49) An appropriate ratio (1.5:1) of Br-DPPZ, [Ru(bpy).sub.2Cl.sub.2].2H.sub.2O and a 30 ml mixed solvent of ethylene glycol and water (V.sub.ethylene glycol:V.sub.water=9:1) were added into a 50 ml three-necked flask, heating and refluxing for reaction for 6 hours (T=120° C.). After the reaction stopped, the reaction system was cooled to room temperature, 80 ml water was added for dilution, and an orange red clear filtrate was obtained by filtration. An excess of NaClO.sub.4 was added to the filtrate, so as to produce a large amount of orange red precipitate; after vacuum filtration, the precipitate was dried in a vacuum drier to obtain an orange yellow crude product. The crude product was dissolved in a small amount of acetonitrile, and purified and separated by a neutral alumina column, packed in column with acetonitrile. Appropriate ratio of acetonitrile was used to elute down a main red component, which was then subjected to rotary evaporation under reduced pressure and drying in a vacuum drier, so as to obtain red crystals with a yield of 80%.
Example 3. Synthesis of RM1601
(50) ##STR00025##
(51) [Ru(bpy).sub.2(BrDPPZ)].sup.2+ (125 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and phenylacetylene (70 μL, 0.637 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 4 bands being separated, wherein the first band was light yellow, the second band was dark reddish brown, the third band was light yellow, and the fourth band was dark reddish brown. Eluting with pure acetonitrile, the first and second bands were collected; eluting with a solvent of acetonitrile:methanol=50:1, the third band was collected, which was a target band; eluting with methanol, the fourth band was obtained. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 31%.
Example 4. Synthesis of RM1602
(52) ##STR00026##
(53) [Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and propargyl alcohol (40 μL, 0.677 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:methanol=20:1, thereby 4 bands being separated, wherein the third band was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 20%.
Example 5. Synthesis of RM1603
(54) ##STR00027##
(55) [Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and 3-aminophenylacetylene (75 μL, 0.666 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 3 bands being separated; eluting with a solvent of acetonitrile:methanol=30:1, the second band was collected, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 35%.
Example 6. Synthesis of RM1604
(56) ##STR00028##
(57) [Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and trimethylsilylacetylene (95 μL, 0.672 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:toluene=1:1, thereby 4 bands being separated, eluting to collect the first band, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 32%.
Example 7. Synthesis of L Ru(bpy).SUB.2.(Br-DPPZ)
(58) ##STR00029##
(59) An appropriate ratio (1.5:1) of Br-DPPZ, L-[Ru(bpy).sub.2(py).sub.2][o,o′-dibenzoyl-L-tartrate].12H.sub.2O and a 30 ml mixed solvent of ethylene glycol and water (V.sub.ethylene glycol:V.sub.water=9:1) were added into a 50 mL three-necked flask, heating and refluxing for reaction for 6 hours (T=120° C.). After the reaction stopped, the reaction system was cooled to room temperature, 80 ml water was added for dilution, and an orange red clear filtrate was obtained by filtration. An excess of NaClO.sub.4 was added to the filtrate, so as to produce a large amount of orange red precipitate; after vacuum filtration, the precipitate was dried in a vacuum drier to obtain an orange yellow crude product. The crude product was dissolved in a small amount of acetonitrile, and purified and separated by a neutral alumina column, packed in column with acetonitrile. The acetonitrile was used to elute down a main red component, which was then subjected to rotary evaporation under reduced pressure and drying in a vacuum drier, so as to obtain red crystals with a yield of 30%.
Example 8. Synthesis of D Ru(bpy).SUB.2.(Br-DPPZ)
(60) ##STR00030##
(61) An appropriate ratio (1.5:1) of Br-DPPZ, D-[Ru(bpy).sub.2(py).sub.2][o,o′-dibenzoyl-L-tartrate].12H.sub.2O and a 30 ml mixed solvent of ethylene glycol and water (V.sub.ethylene glycol:V.sub.water=9:1) were added into a 50 mL three-necked flask, heating and refluxing for reaction for 6 hours (T=120° C.). After the reaction stopped, the reaction system was cooled to room temperature, 80 ml water was added for dilution, and an orange red clear filtrate was obtained by filtration. An excess of NaClO.sub.4 was added to the filtrate, so as to produce a large amount of orange red precipitate; after vacuum filtration, the precipitate was dried in a vacuum drier to obtain an orange yellow crude product. The crude product was dissolved in a small amount of acetonitrile, and purified and separated by a neutral alumina column, packed in column with acetonitrile. An appropriate ratio of acetonitrile was used to elute down a main red component, which was then subjected to rotary evaporation under reduced pressure and drying in a vacuum drier, so as to obtain red crystals with a yield of 40%.
Example 9. Synthesis of LRM1601
(62) ##STR00031##
(63) L[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and phenylacetylene (70 μL, 0.637 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 4 bands being separated, wherein the first band was light yellow, the second band was dark reddish brown, the third band was light yellow, and the fourth band was dark reddish brown. Eluting with pure acetonitrile, the first and second bands were collected; eluting with a solvent of acetonitrile:methanol=50:1, the third band was collected, which was a target band; eluting with methanol, the fourth band was obtained. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 35%.
Example 10. Synthesis of DRM1601
(64) ##STR00032##
(65) D[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and phenylacetylene (70 μL, 0.637 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 4 bands being separated, wherein the first band was light yellow, the second band was dark reddish brown, the third band was light yellow, and the fourth band was dark reddish brown. Eluting with pure acetonitrile, the first and second bands were collected; eluting with a solvent of acetonitrile:methanol=50:1, the third band was collected, which was a target band; eluting with methanol, the fourth band was obtained. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 50%.
Example 11. Synthesis of LRM1602
(66) ##STR00033##
(67) L[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and propargyl alcohol (40 μL, 0.677 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:methanol=20:1, thereby 4 bands being separated, wherein the third band was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 12%.
Example 12. Synthesis of DRM1602
(68) ##STR00034##
(69) D[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and propargyl alcohol (40 μL, 0.677 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:methanol=20:1, thereby 4 bands being separated, wherein the third band was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 15%.
Example 13. Synthesis of LRM1603
(70) ##STR00035##
(71) L[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and 3-aminophenylacetylene (75 μL, 0.666 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 3 bands being separated; eluting with a solvent of acetonitrile:methanol=30:1, the second band was collected, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 36%.
Example 14. Synthesis of DRM1603
(72) ##STR00036##
(73) D[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and 3-aminophenylacetylene (75 μL, 0.666 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 3 bands being separated; eluting with a solvent of acetonitrile:methanol=30:1, the second band was collected, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 37%.
Example 15. Synthesis of LRM1604
(74) ##STR00037##
(75) L[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and trimethylsilylacetylene (95 μL, 0.672 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:toluene=1:1, thereby 4 bands being separated, eluting to collect the first band, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 33%.
Example 16. Synthesis of DRM1604
(76) ##STR00038##
(77) D[Ru(bpy).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and trimethylsilylacetylene (95 μL, 0.672 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:toluene=1:1, thereby 4 bands being separated, eluting to collect the first band, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 35%.
Example 17. Synthesis of [Ru(phen).SUB.2.(Br-DPPZ)].SUP.2+
(78) ##STR00039##
(79) An appropriate ratio (1.5:1) of Br-DPPZ, [Ru(phen).sub.2Cl.sub.2].2H.sub.2O and a 30 ml mixed solvent of ethylene glycol and water (V.sub.ethylene glycol:V.sub.water=9:1) were added into a 50 mL three-necked flask, heating and refluxing for reaction for 6 hours (T=120° C.). After the reaction stopped, the reaction system was cooled to room temperature, 80 ml water was added for dilution, and an orange red clear filtrate was obtained by filtration. An excess of NaClO.sub.4 was added to the filtrate, so as to produce a large amount of orange red precipitate; after vacuum filtration, the precipitate was dried in a vacuum drier to obtain an orange yellow crude product. The crude product was dissolved in a small amount of acetonitrile, and purified and separated by a neutral alumina column, packed in column with acetonitrile. An appropriate ratio of acetonitrile was used to elute down a main red component, which was then subjected to rotary evaporation under reduced pressure and drying in a vacuum drier, so as to obtain red crystals with a yield of 60%.
Example 18. Synthesis of RPM1601
(80) ##STR00040##
(81) [Ru(phen).sub.2(BrDPPZ)].sup.2+ (125 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and phenylacetylene (70 μL, 0.637 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, the first band was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 33%.
Example 19. Synthesis of RPM1602
(82) ##STR00041##
(83) [Ru(phen).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and propargyl alcohol (40 μL, 0.677 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:methanol=20:1, thereby 4 bands being separated, wherein the third band was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 20%.
Example 20. Synthesis of RPM1603
(84) ##STR00042##
(85) [Ru(phen).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and 3-aminophenylacetylene (95 μL, 0.672 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with pure acetonitrile, thereby 3 bands being separated; eluting with a solvent of acetonitrile:methanol=30:1, the second band was collected, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 45%.
Example 21. Synthesis of RPM1604
(86) ##STR00043##
(87) [Ru(phen).sub.2(BrDPPZ)].sup.2+ (130 mg, 0.125 mmol) was weighed and taken into a 10 ml microwave reaction tube (with a stirrer therein), and 5 ml anhydrous acetonitrile was added thereto, ultrasonic processing for complete dissolution. Afterwards, under the protection of nitrogen, appropriate amount of triethylamine (70 μL, 0.501 μmol) and trimethylsilylacetylene (95 μL, 0.672 μmol) were added, and then the catalyst Pd(PPh.sub.3).sub.2Cl.sub.2 (17.6 mg) and CuI (20 mg) were quickly added; after being airtight, it was put into a microwave reactor for microwave assisted heating at 140° C. for 30 min. After the completion of the reaction, the reaction system was cooled to room temperature, and was subjected to filtration to obtain a filtrate. The filtrate was purified by a neutral alumina column, eluting with a solvent of acetonitrile:toluene=1:1, thereby 4 bands being separated, eluting to collect the second band, which was a target band. A solution of the target band was subjected to vacuum concentration to obtain a reddish brown solid, which was the target compound, with a yield of 32%.
Example 22. Inhibitory Effect of the Ruthenium Complex Containing Alkynyl Group on MDA-MB-231 Cells
(88) The Ruthenium complex containing alkynyl group in 1 mM stock solution was dissolved in DMSO, cell viability was measured by measuring the ability of cells to convert MTT into purple formazan dye. The cells were seeded in a 96-well plate for tissue culture for 24 hours, and then the cells were incubated with different concentrations of the complexes for 72 hours; at a higher temperature, 20 mL/well of MTT solution (5 mg/mL phosphate buffered saline) was added, and incubated for 5 hours. A microplate spectraphotometer (SpectroAmaxt250) was used to measure the color intensity of the formazan solution that reflects the cell growth conditions at 570 nm. The inhibitory effect of different Ruthenium complexes containing alkynyl group on MDA-MB-231 cells are shown in Table 1, wherein DRM1601 and DRM1603 have a good inhibitory effect on breast cancer cell line MDA-MB-231, especially the IC.sub.50 value of DRM1603 to MDA-MB-231 reaches 3.5±0.14 μM.
(89) TABLE-US-00001 TABLE 1 Inhibitory effect of different Ruthenium complexes containing alkynyl group on MDA-MB-231 cells IC.sub.50/μM Comp. MDA-MB-231 LRM1601 35.95 ± 1.6 LRM1602 382.5 ± 23.sup. LRM1603 53.5 ± 1.7 DRM1601 9.8 ± 0.2 DRM1602 162.6 ± 10.5 DRM1603 3.5 ± 0.14
Example 23. A Use of the Ruthenium Complex Containing Alkynyl Group as a Fluorescence Probe
(90) In a complete growth medium with 5×10.sup.4 cells per ml, MDA-MB-231 cells were incubated with RM1601 (20 μM) at 37° C. for 72 hours. The cells were washed for three times with phosphate buffered saline (PBS), fixed with 4% paraformaldehyde, and labeled with 4′,6-diamidino-2-phenylindole (DAPI; 0.5 μg/mL). A laser scanning confocal microscope was used to observe the fluorescence property of compound RM1601 and its localization in the cells; the results are shown in
(91) Specific embodiments of the present invention have been described above, but the specific embodiments are only used as examples and should not be taken as limitation. Those skilled in the art should be able to understand that any equivalent modifications and substitutions to the present invention are within the scope of the present invention. Therefore, without departing from the spirit and the scope of the present invention, a variety of equivalent changes and modifications should be within the scope of the present invention.