CN105622674A - Tetravalent platinum complex with bioactive group and preparation method of tetravalent platinum complex - Google Patents

Abstract

The invention discloses a tetravalent platinum complex with a bioactive group and a preparation method of the tetravalent platinum complex. The tetravalent platinum complex is a platinum (IV) complex and has the structure shown in the formula II (please see the formula in the description), wherein in the formula II, Y is OH or Cl, and Bio represents the bioactive group. The platinum (IV) complex is prepared according to the equation in the formula III (please see the formula in the description), wherein in the formula III, Y is OH or Cl, Bio-OH represents a compound with bioactivity, TBTU represents a coupling agent O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium tetrafluoroborate, TEA represents a catalyst triethylamine, DMF represents solvent N,N-dimethyl formamide, and DMSO represents solvent dimethylsulfoxide. Cis-platinum is adopted for the bottom face of an octahedron, a small-molecular targeted or medicine active group is introduced to one axial position, a hydroxyl group or helium atom is introduced into another axial position, and the anti-tumor tetravalent platinum complex overcoming cisplatin resistance is provided, so the high-efficiency and low-toxin platinum (IV) complex is obtained.

Description

One class tetravalence platinum complex containing bio-active group and preparation method thereof

Technical field

The present invention relates to antitumor tetravalence platinum complex, it is specifically related to the bottom surface with cisplatin skeleton for tetravalence platinum complex octahedral structure, introduce at an axial location and single there is bioactive group, introduce hydroxyl or chlorine atom at another axial location, obtain a class and there is platinum (IV) coordination compound of anti-tumor activity; The invention still further relates to the preparation method of this platinum complexes and application.

Background technology

The antitumor bivalence platinum medicines such as cisplatin, carboplatin and oxaliplatin have been widely used in clinic, for treating relevant cancer. As first metal antitumor drug used clinically, cisplatin mainly acts on the guanine N7 position target spot of DNA, by being cross-linked to form adduct with DNA, makes tumor cell generation apoptosis, causes cells arrest and cell death. Big quantity research shows, cisplatin trigger cancer cells death mainly includes four-stage: (1) transmembrane transport: produce cellular uptake by the mode of actively diffusion and Passive intake; (2) hydration dissociation: due to the concentration difference of intraor extracellular chloride ion, it is easy to form platinum (II) hydrated cation; (3) targeting migrates: platinum (II) ion forms Pt-DNA adduct with DNA; (4) DNA damage is lethal: by acting on DNA, causes the death of cancerous cell.

According to above-mentioned mechanism, along platinum medicine after intravenous injection, before arriving tumor cell, can with the such as glutathion effect poisoning and deactivation of the nucleophile in blood. Therefore, existing exist some major defects along platinum medicine: one is show corresponding toxicity, mainly Toxicity of Kidney and bone marrow toxicity; Two is the drug resistance produced after medication. These deficiencies limit the application of these platinum (II) medicine to a certain extent.

Research in recent years finds, platinum (IV) coordination compound has good stability, can reduce and the reaction of internal nucleophile, introduce certain axial group and can regulate platinum (IV) coordination compound hydrophilic and lipotropy, promote the absorption of medicine. Platinum (IV) ion generates after platinum (II) ion through reduction in vivo, can with cancerous cell DNA effect, if at platinum (IV) coordination compound axially introduced into bio-active group, then be likely to and platinum (II) ion produce synergistic antitumor activity. Therefore, platinum (IV) coordination compound arouses widespread concern as anti-tumor predrug.

The structure activity relationship of platinum (IV) coordination compound is shown in Fig. 1, mainly includes the following aspects: the kinetics of X leaving group part major effect compound, toxicity, dissolubility; Am carrier ligand major effect Pt-DNA adduction mode and anti-tumor activity; The ability of the targeting of Y-axis part major decision medicine, biological activity and entrance tumor cell, affects the redox property of compound, the ability being namely reduced in the tumor tissues of hypoxia. Therefore, the targeting of medicine can be increased and fat-soluble by introducing functional type axial ligand, overcome toxic and side effects, increase the anti-tumor activity of compound.

Although people have carried out a few thing in platinum (IV) drug research, design and be prepared for the coordination compound of multiple type, it is desirable with the space structure of platinum (IV) coordination compound, it is thus achieved that high-efficiency low-toxicity and suitable platinum medicine are without crossing drug resistant or the medicine with selective therapy effect. But up to now, there is presently no and be capable of targeting or overcome platinum (IV) coordination compound of cisplatin resistance to list as medicine.

Summary of the invention

Technical problem: it is an object of the invention to utilize the space structure of platinum (IV) coordination compound, adopting cisplatin is octahedra bottom surface, little molecular targeted or pharmaceutically active group is introduced at an axial location, another axial location introduces hydroxyl or chlorine atom, there is provided and there is the antitumor tetravalence platinum complex overcoming cisplatin resistance, to obtaining platinum (IV) medicine of high-efficiency low-toxicity; The preparation method that present invention also offers these platinum (IV) coordination compound, and they are in the application prepared on antitumor drug.

Technical scheme: the present invention is the tetravalence platinum complex that a class contains bio-active group, described tetravalence platinum complex and platinum (IV) coordination compound, its structure is such as shown in Formula II:

In Formula II, Y is OH or Cl; Bio represents bio-active group.

The preparation method that one class of the present invention contains the tetravalence platinum complex of bio-active group, described platinum (IV) coordination compound, the reaction equation shown in formula III carries out,

In formula III, Y is OH or Cl; Bio-OH represents has bioactive compound, and TBTU represents coupling reagent O-BTA-N, N, N ', N '-tetramethylurea Tetrafluoroboric acid, and TEA represents catalyst of triethylamine, and DMF represents solvent DMF, and DMSO represents solvent dimethyl sulfoxide;

Specifically adopt with the following method: equimolar reactant B io-OH and coupling reagent TBTU is mixed in dry DMF or DMSO, under stirring at room temperature, add equimolar TEA, it is subsequently adding equimolar Pt (IV) reactant A or B, reactant liquor stirs 12-48 hour under spending at 30-60 DEG C under nitrogen protection, then removal of solvent under reduced pressure, and concentrated solution separates through silica gel column chromatography, eluent is dichloromethane and methanol mixed solvent, obtains yellow solid product;

The structure such as Formulas I V of Pt (IV) reactant A and B represents,

Wherein A represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], reacted in water by cisplatin and hydrogen peroxide and prepare; B represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], reacted in water by cisplatin and chlorosuccinimide and prepare.

Described Bio-OH represents has bioactive compound structure shown in Formulas I (a-f),

Wherein, (a) is the anti-tumor drugs targeting silmitasertib of known entrance clinical trial, is a kind of Protein kinase C K2 inhibitor, referred to as CX-OH; B () is known marketed drug chlorambucil, be a kind of target spot be the antitumor drug of DNA, referred to as CHL-OH; C () is biotin, i.e. biotin, referred to as BT-OH; D () is the carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUA-OH; E () is the triazole carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUT-OH; F () is the carboxylic acid derivates of known antitumor drug SAHA, be a kind of PARP inhibitor, referred to as SAA-OH.

The preparation of described CUT-OH is undertaken by the reaction equation shown in Formula V,

Preparation CUT-OH adopts and is made by method:

L) sodium acetate of equimolar 2,4-4-dihydroxy benzaldehydes and acetoglycocoll and triplication is added in acetic anhydride, be stirred at reflux 4h, reactant liquor is cooled to room temperature, pours in frozen water, obtain yellow mercury oxide, filtering, wash twice with frozen water, drying solid obtains intermediate M1;

2) M1 being added concentrated hydrochloric acid: ethanol is in the mixed solution of 2: 1, reflux 1h, and reactant liquor is cooled to room temperature, pour in frozen water, then under condition of ice bath, it is slowly added to the sodium nitrite of qdx, after adding stirring 15min, it is slow added into the Hydrazoic acid,sodium salt of triplication, continue stirring 15min, filter out tan precipitate, washing, column chromatography for separation obtains intermediate M2, and eluent is petroleum ether: ethyl acetate is 5: 1 mixed solvents;

3) M2 is suspended in methanol, add equimolar fourth-3-alkynes-1-alcohol and a small amount of sodium ascorbate, 10min is stirred at room temperature under nitrogen protection, again the copper sulphate pentahydrate being dissolved in the qdx of water is joined in reactant liquor, room temperature lucifuge stirring 4h, then rotation is evaporated off the methanol in dereaction liquid, add water precipitation yellow solid, filter, gained solid pure water twice, dry to obtain intermediate M3;

4) M3 being dissolved in DMF, add the succinic anhydride of qdx and the DMAP of catalytic amount, 50 DEG C of reactions are overnight, it is cooled to room temperature, concentration of reaction solution, column chromatography for separation obtains light yellow product, and eluent is petroleum ether: ethyl acetate: methanol is 10: 10: 1 mixed solvents.

Beneficial effect:

(1), anti tumor activity in vitro

With cisplatin resistance stomach cancer cell SGC-7901/CCCP, brain glioblastoma cell U87 or neurogliocytoma cell U251, prepared compound human lung cancer cell A549, colon cancer cell HCT-116, hepatocellular carcinoma H22, breast cancer cell MCF-7 and MDA-MB-231, ovarian cancer cell A2780, pancreatic cancer cell PANC-1, stomach cancer cell SGC-7901 have been carried out anti tumor activity in vitro evaluation, and cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations.

The experimental data of the compound 1 and 2 containing target protein kinase CK2 inhibitor group CX is in Table 2. Compound 1 to the cancerous cell MCF-7 of CK2 high expressed, HepG2, HCT-116 and A2780 activity be better than cisplatin 2.5-4.5 times, the cancerous cell SGC-7901 express CK2 moderate or the suppression ratio of the cancerous cell PANC-1 of low expression are close with cisplatin, it should be noted that cisplatin resistance cancerous cell SGC-7901/CCCP is significantly inhibited effect by this compound, drug-resistance factor is 0.60. Compound 2 is high to CK2, the activity of the cancerous cell of middle expression is slightly less than cisplatin, but cisplatin resistance cancerous cell SGC-7901/CCCP is also illustrated that significant inhibitory action by it, and drug-resistance factor reaches 0.24. Table 2 data show that the introducing of targeting group makes platinum (IV) coordination compound be significantly improved compared with the anti-tumor activity of cisplatin, and axially the activity of the axial compound 2 containing chlorine atom of compound 1 ratio containing oh group is better.

The experimental data of the compound 3 containing the CHL group that target spot is DNA is in Table 3. This compound is suitable with cisplatin to the activity of surveyed cancerous cell, and cisplatin resistance cancerous cell SGC-7901/CCCP is had certain inhibitory action by it, and drug-resistance factor is 2.65.

The experimental data of the compound 4 and 5 containing biotin BT group is in Table 4. The activity of the compound 4 two cancerous cell to surveying is below cisplatin, but compound 5 is more sensitive to hepatocellular carcinoma H22 and stomach cancer cell SGC701, cytotoxic activity is better than cisplatin, particularly that cisplatin resistance cancerous cell SGC-7901/CCCP is particularly sensitive, its activity is 21 times of cisplatin, and drug-resistance factor is 0.42. Result shows that the compound 4 axially containing oh group is more weak than the activity of the compound 5 axially containing chlorine atom.

The experimental data of the compound 6 containing coumarin carboxylic acid derivates CUA group is also in Table 4. The activity of the cancerous cell of all tests is all higher than cisplatin by compound 6, as for the cytotoxic activity of colon cancer cell HCT116 being 30 times of cisplatin, it is 23 times of cisplatin to the cytotoxic activity of stomach cancer cell SGC-7901, it is 19 times of cisplatin to the cytotoxic activity of cisplatin resistance stomach cancer cell SGC-7901/CCCP, but its drug-resistance factor is 5.31.

The experimental data of the compound 7 of the triazole carboxylic acid derivates CUT group containing coumarin is in Table 5. The activity of human glioma cell U87 and neurogliocytoma cell U251 is better than cisplatin by compound 7, and cytotoxic activity is 2-3 times of cisplatin.

The experimental data of the compound 8 containing PARP inhibitor SAA group is in Table 6. The activity of the compound 8 cancerous cell to testing is suitable with cisplatin, and what have is slightly strong, and what have is slightly weak, but it is to 11 times that the cytotoxic activity of cisplatin resistance stomach cancer cell SGC-7901/CCCP is cisplatin, and drug-resistance factor is 0.66.

Result above shows, introduces bio-active group and shows effective antitumour activity to platinum (IV) coordination compound that cisplatin is parent.

(2), anti-tumor in vivo activity

With prepared compound 1, the growth of SGC-7901 cells nude mouse xenograft tumor and effect anti-tumor in vivo activity rating, cisplatin and CX-OH and drug combination thereof are carried out as comparison.

Experimental result is in Table 7 (inhibitory action that SGC-7901 cells nude mouse xenograft tumor is grown by given the test agent), Fig. 2 is that SGC-7901 cells nude mouse xenograft tumor is grown the impact of change in volume by given the test agent, Fig. 3 is the inhibitory action that SGC-7901 cells nude mouse xenograft tumor is grown by given the test agent, and Fig. 4 is the given the test agent impact on SGC-7901 cells nude mouse xenograft tumor nude mice body weight. From the data in table 7, it can be seen that the tumour inhibiting rate of compound 1 is far above isodose cisplatin and CX-OH. Compared with cisplatin and CX-OH drug combination, the CX-OH drug combination that the tumour inhibiting rate of compound 1 is administered in conjunction with the low frequency higher than cisplatin, the CX-OH drug combination being administered in conjunction with high frequency time with cisplatin is suitable. As shown in Figures 2 and 3, being no matter volume or the weight data of transplanted tumor, the CX-OH drug combination that compound 1 is administered in conjunction with high frequency time with cisplatin can significantly inhibit the growth of nude mouse xenograft tumor, higher than other test group. It should be noted that Fig. 4, compound 1 on animal subject body weight almost without impact, and cisplatin and the weight of animals impact is relatively big with CX-OH drug combination, illustrate that the toxicity of compound 1 is less, there is the feature of high-efficiency low-toxicity.

Accompanying drawing explanation

Fig. 1. the structure activity relationship of platinum (IV) coordination compound.

Fig. 2. SGC-7901 cells nude mouse xenograft tumor is grown the impact of change in volume by given the test agent.

Fig. 3. the inhibitory action that SGC-7901 cells nude mouse xenograft tumor is grown by given the test agent.

Fig. 4. the given the test agent impact on SGC-7901 cells nude mouse xenograft tumor nude mice body weight.

Detailed description of the invention

Targeting involved in the present invention or pharmaceutically active group are known have bioactive compound or derivatives thereof, their structure shown in Formulas I (a-f),

Wherein, the anti-tumor drugs targeting (silmitasertib) that (a) is known entrance clinical trial, is a kind of Protein kinase C K2 inhibitor, referred to as CX-OH; B () is known marketed drug chlorambucil, be a kind of target spot be the antitumor drug of DNA, referred to as CHL-OH; C () is biotin, i.e. biotin (in many tumor cells biotin acceptor process LAN), referred to as BT-OH; D () is the carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUA-OH; E () is the triazole carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUT-OH; F () is the carboxylic acid derivates of known antitumor drug SAHA, be a kind of PARP inhibitor, referred to as SAA-OH.

Platinum of the present invention (IV) coordination compound, its structure is such as shown in Formula II:

In Formula II, Y is OH or Cl; Bio represents bio-active group, for parent CX, BT, CHL, CUA, CUT or SAA of compound structure shown in Formulas I (a-f); The structure of concrete representative compound is as shown in table 1:

The structure of table 1. representative compound 1.8

Preparation platinum (IV) coordination compound shown in Formula II, the reaction equation according to formula III carries out,

In formula III, Y is OH or Cl; Compound shown in Bio-OH representative formula I (a-f), such as CX-OH, BT-OH, CHL-OH, CUA-OH, CUT-OH or SAA-OH; TBTU represents coupling reagent O-BTA-N, N, N ', N '-tetramethylurea Tetrafluoroboric acid, and TEA represents catalyst of triethylamine, and DMF represents solvent DMF, and DMSO represents solvent dimethyl sulfoxide.

Preparation platinum (IV) coordination compound described in Formula II; employing is made by method: the equimolar reactant B io-OH and the coupling reagent TBTU dry DMF in certain volume or DMSO are mixed; under stirring at room temperature; add equimolar TEA; it is subsequently adding equimolar Pt (IV) reactant A or B; reactant liquor stirs 12-48 hour under spending at 30-60 DEG C under nitrogen protection; then removal of solvent under reduced pressure; concentrated solution separates through silica gel column chromatography; eluent is dichloromethane and methanol mixed solvent, obtains yellow solid product.

The structure such as Formulas I V of Pt (IV) reactant A and B represents,

Wherein A represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], reacted in water by cisplatin and hydrogen peroxide by literature method (Can.J.Chem.1982,60,529) and prepare; B represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], reacted in water by cisplatin and chlorosuccinimide by literature method (Inorg.Chem.2014,53,9326) and prepare.

In reactant B io-OH, and CX-OH (J.Med.Chem., 2011,54,635), CHL-OH (Anti-cancerDrugs, 2006,17,511), CUA-OH (ColloidPolym.Sci.2014,292,85) and SAA-OH (J.Med.Chem., 2010,53,1937) prepared by the method with reference to corresponding bibliographical information; BT-OH is commodity.

The preparation of CUT-OH is undertaken by the reaction equation shown in Formula V,

Preparation CUT-OH adopts and is made by method:

1. the sodium acetate of equimolar 2,4-4-dihydroxy benzaldehydes and acetoglycocoll and triplication is added in the acetic anhydride of certain volume, be stirred at reflux 4h, reactant liquor is cooled to room temperature, pours in frozen water, obtain yellow mercury oxide, filtering, wash twice with frozen water, drying solid obtains intermediate M1;

2. being added by M1 in the mixed solution of concentrated hydrochloric acid and ethanol (2: 1), reflux 1h, and reactant liquor is cooled to room temperature, pour in frozen water, then under condition of ice bath, it is slowly added to the sodium nitrite of qdx, after adding stirring 15min, it is slow added into the Hydrazoic acid,sodium salt of triplication, continue stirring 15min, filter out tan precipitate, washing, column chromatography for separation obtains intermediate M2, and eluent is petroleum ether and ethyl acetate (5: 1) mixed solvent;

3. a certain amount of M2 is suspended in methanol, add equimolar fourth-3-alkynes-1-alcohol and a small amount of sodium ascorbate, 10min is stirred at room temperature under nitrogen protection, again the copper sulphate pentahydrate being dissolved in the qdx of a small amount of water is joined in reactant liquor, room temperature lucifuge stirring 4h, then rotation is evaporated off the methanol in dereaction liquid, add water precipitation yellow solid, filter, gained solid pure water twice, dry to obtain intermediate M3;

4. a certain amount of M3 being dissolved in DMF, adds the succinic anhydride of qdx and the DMAP of catalytic amount, 50 DEG C of reactions are overnight, it is cooled to room temperature, concentration of reaction solution, column chromatography for separation obtains light yellow product, and eluent is petroleum ether, ethyl acetate and methanol (10: 10: 1) mixed solvent.

Platinum (IV) coordination compound prepared by the inventive method determines the molecular structure of compound through nucleus magnetic hydrogen spectrum and mass spectrum, and some compounds have also characterized through nuclear-magnetism carbon spectrum. In compound mass spectrum, all there are multiple isotopic peaks of platinum element in quasi-molecular ion peak.

Shown in table 1, some tumor cells are had significant inhibitory action by representative platinum (IV) coordination compound, and what have exceedes cisplatin, and the compound particularly having, to cisplatin resistance cancerous cell performance sensitivity, has the ability overcoming cisplatin resistance. These cancerous cell include pulmonary carcinoma, colon cancer, hepatocarcinoma, breast carcinoma, ovarian cancer, cancer of pancreas, gastric cancer, cerebral glioma and neurogliocytoma cell. The in vivo test of compound 1 shows, the tetravalence platinum complex containing targeted drug group shows the anti-tumor activity of high-efficiency low-toxicity, can be applicable to prepare cancer therapy drug.

The present invention is by following embodiment further instruction, but these explanations are not limiting as the present invention. Unless otherwise indicated, some reactants include suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂] and suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl] all adopt literature method to prepare.

(1), the preparation of compound

The preparation of embodiment 1. compound 1

157.4mg (0.45mmol) CX-OH and 144.5mg (0.45mmol) TBTU is dissolved in 15mL dry DMF, 10min is stirred at room temperature, it is subsequently adding 45.5mg (0.45mmol) TEA, after continuing stirring 5min, add 150.0mg (0.45mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], the lower 50 DEG C of reaction 48h of nitrogen protection. Concentration of reaction solution, concentrated solution separates through silica gel column chromatography, and eluent is dichloromethane and methanol mixed solvent (10: 1), obtains yellow product 79.9mg, productivity 40%.

¹HNMR (DMSO-d6, ppm): �� 4.23 (s, 6H), 7.13 (d, J=7.9,1H), 7.44 (t, J=8.0,1H), 7.97 (d, J=8.4,1H), 8.07 (d, J=8.6,1H), 8.27 (m, 2H), 8.46 (t, J=2.0,1H), 8.61 (d, J=5.9,1H), 8.82 (d, J=8.5,1H), 9.14 (d, J=5.9,1H), 9.7 (s, 1H), 10.2 (s, 1H).¹³CNMR (DMSO-d6, ppm): �� 116.73,119.53,120.53,122.58,122.75,122.92,124.37,124.54,127.26,128.69,130.46,132.10,142.27,143.66,147.92,148.10,150.46,167.57.ESI-MS:[M-H]⁺=665.1.

The preparation of embodiment 2. compound 2

Suitable with reactant, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl] substitute suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂] prepare with reference to method described in embodiment 1, obtain yellow product, productivity 38%.

¹HNMR (DMSO-d6, ppm): �� 6.38 (s, 6H), 7.16 (d, J=8.1,1H), 7.49 (t, J=8.2,1H), 7.99 (d, J=8.4,1H), 8.08 (d, J=8.4,1H), 8.3 (m, 2H), 8.61 (d, J=5.6,1H), 8.85 (d, J=8.5,1H), 9.01 (d, J=5.6,1H), 9.7 (s, 1H), 10.2 (s, 1H).¹³CNMR (DMSO-d6, ppm): �� 116.87,119.53,120.52,121.85,122.29,122.60,124.28,125.38,127.58,128.77,130.62,133.28,135.44,142.40,143.48,147.77,148.22,150.41,173.19.ESI-MS:[M-H]⁺=681.9

The preparation of embodiment 3. coordination compound 3

100.0mg (0.33mmol) CHL-OH is dissolved in 5mL dry DMF, add 105.6mg (0.33mmol) TBTU, 5min is stirred at room temperature, it is subsequently adding 33.3mg (0.33mmol) TEA, after continuing to be stirred for 10min, add 115.8mg (0.33mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], room temperature reaction 24h under nitrogen protection. Concentration of reaction solution, concentrated solution separates through silica gel column chromatography, and eluent is dichloromethane and methanol mixed solvent (5: 1), obtains yellow solid 95.7mg, and productivity is 46%.

¹H-NMR (DMSO-d6, ppm): �� 1.67-1.72 (2H, m), 2.19-2.24 (2H, m), 2.45 (2H, m), 3.69 (8H, s), 6.02-6.35 (6H, m), and 6.64-6.67 (2H, d), 7.02-7.05 (2H, d) .ESI-MS:[M+H]⁺=637.9 (100%).

The preparation of embodiment 4. coordination compound 4

122.1mg (0.50mmol) BT-OH and 160.5mg (0.50mmol) TBTU is dissolved in the anhydrous DMSO of 10mL, 15min is stirred at room temperature, it is subsequently adding 50.6mg (0.50mmol) TEA, after continuing stirring 15min, add 160.7mg (0.50mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], the lower 60 DEG C of reaction 36h of nitrogen protection. Concentration of reaction solution, concentrated solution separates through silica gel column chromatography, and eluent is dichloromethane and methanol mixed solvent (5: 1), obtains yellow solid 50.0mg, productivity 18%.

¹HNMR (DMSO-d6, ppm): �� �� 1.34-1.48 (m, 6H), 2.18 (t, 2H), 2.60 (s, 1H), 2.80 (m, 1H), 3.10 (s, 1H), 4.14 (s, 1H), 4.31 (s, 1H), 5.94 (m, 6H), 6.32 (d, 2H) .ESI-MS:[M-H]-=559.1, [M+Cl]^-=595.0.

The preparation of embodiment 5. coordination compound 5

134.0mg (0.55mmol) BT-OH and 176.6mg (0.55mmol) TBTU is dissolved in the anhydrous DMSO of 10mL, 15min is stirred at room temperature, it is subsequently adding 55.7mg (0.55mmol) TEA, after continuing stirring 15min, add 193.9mg (0.55mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], 40 DEG C of reaction 24h under nitrogen protection. Concentration of reaction solution, concentrated solution separates through silica gel column chromatography, and eluent is dichloromethane and methanol mixed solvent (5: 1), obtains yellow solid 100.1mg,

Productivity: 31%.

¹HNMR (DMSO-d6, ppm): �� 1.62-1.35 (m, 6H), 2.23 (s, 2H), 2.57 (m, 3H), 2.81,3.11,4.15 (s, 1H), 4.31 (s, 1H), 6.18-6.36 (m, 8H).¹³CNMR (DMSO-d6, ppm): 825.01,25.98,28.66,33.97,36.62,55.93,59.66,61.48,163.22,181.05.ESI-MS:[M-H]^-=577.0.

The preparation of embodiment 6. coordination compound 6

110.0mg (0.50mmol) CUA-OH and 160.5mg (0.50mmol) TBTU is dissolved in the anhydrous DMSO of 20mL, 15min is stirred at room temperature, it is subsequently adding 50.6mg (0.50mmol) TEA, after continuing stirring 15min, add 160.7mg (0.50mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], 60 DEG C of reaction 48h under nitrogen protection. Concentration of reaction solution, concentrated solution separates through silica gel column chromatography, and eluent is dichloromethane and methanol mixed solvent (5: 1), obtains yellow solid 50.0mg, productivity: 19%.

¹HNMR (DMSO-d6, ppm): �� 1.24 (s, 1H), 4.65 (s, 2H), 5.95 (m, 6H), 6.28 (d, 1H), 6.94 (m, 2H), 7.60 (d, 1H), 7.99 (d, 1H) .ESI-MS:[M-H]^-=535.0.

The preparation of embodiment 7. coordination compound 7

201.6mg (0.54mmol) CUT-OH and 173.4mg (0.54mmol) TBTU is dissolved in 30mL dry DMF, 10min is stirred at room temperature, it is subsequently adding 54.5mg (0.54mmol) TEA, after continuing stirring 10min, add 180.4mg (0.54mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], the lower 50 DEG C of reaction 36h of nitrogen protection. After reactant liquor rotation is evaporated off solvent, concentrated solution is through column chromatography for separation, and eluent is dichloromethane and methanol mixed solvent (5: 1), obtains yellow solid 90.1mg, productivity: 24%.

¹HNMR (DMSO-d6, ppm): �� 2.45 (4H, s), 3.02-3.05 (2H, m), and 4.28-4.31 (2H, m), 5.82-5.93 (6H, m), 6.30 (1H, s), and 6.46-6.48 (1H, d), 7.39-7.41 (1H, d), 8.25 (1H, s), 8.29 (1H, s) .ESI-MS:[M-H]^-=688.1.

The preparation of embodiment 8. coordination compound 8

77.8mg (0.31mmol) SAA-OH and 100.2mg (0.31mmol) TBTU is dissolved in 15mL dry DMF, 5min is stirred at room temperature, it is subsequently adding 31.6mg (0.31mmol) TEA, after continuing stirring 5min, add 110.2mg (0.31mmol) suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], the lower 50 DEG C of reaction 12h of nitrogen protection. After reactant liquor rotation is evaporated off solvent, concentrated solution is through column chromatography for separation, and eluent is dichloromethane and methanol mixed solvent (15: 1), obtains yellow solid 57.9mg, productivity: 32%.

¹HNMR (DMSO-d6, ppm): �� 1.31 (s, 4H), 1.53 (d, J=28.7Hz, 5H), 2.35-2.18 (m, 4H), 6.18 (s, 7H), 7.01 (t, J=6.9Hz, 1H), 7.28 (t, J=7.6Hz, 2H), 7.58 (d, J=7.8Hz, 2H).¹³CNMR (DMSO-d6, ppm): 825.53,28.95,36.78,39.62,39.76,39.90,40.04,40.18,40.31,40.45,119.52,123.36,129.08,171.73.ESI-MS:[M-H]^-=582.0.

The preparation of embodiment 9.CUT-OH

(1) preparation of M1 and M2

2.76g (20mmol) 2,4-4-dihydroxy benzaldehyde, 2.34g (20mmol) acetoglycocoll and 4.92g (60mmol) sodium acetate are added in 100mL acetic anhydride, is stirred at reflux 4h. It is cooled to room temperature, reactant liquor is poured in frozen water (about 500mL), obtain yellow mercury oxide, filter, wash 2 times with frozen water, obtain M1 crude product.

Being placed in reaction bulb by above-mentioned yellow solid, add the mixed solution of 30mL concentrated hydrochloric acid and ethanol (2: 1), reflux 1h. It is cooled to room temperature, reactant liquor is poured into equipped with in 40mL frozen water. Under Z condition of ice bath, it is slowly added to 2.76g (40mmol) sodium nitrite, add stirring 15min, it is slow added into 3.90g (60mmol) Hydrazoic acid,sodium salt, continues stirring 15min, filter out tan precipitate, washing, column chromatography for separation obtains intermediate M21.51g, and eluent is petroleum ether and ethyl acetate (5: 1) mixed solvent, productivity 37%.

¹H-NMR (DMSO-d6, ppm): �� 6.76 (d, 1H), 6.79-6.82 (dd, 1H), 7.47-7.50 (d, 1H), 7.59 (s, 1H), 10.51 (s, 1H).

(2) preparation of M3

500.0mg (2.46mmol) M2 is suspended in 10mL methanol; 172.3mg (2.46mmol) fourth-3-alkynes-1-alcohol and 24.4mg (0.12mmol) sodium ascorbate; 10min is stirred at room temperature under nitrogen protection; again 122.9mg (0.49mmol) copper sulphate pentahydrate being dissolved in 2mL water is joined in reactant liquor; room temperature lucifuge stirring 4h; then rotation is evaporated off the methanol in dereaction liquid; add 10mL elutriation and go out yellow solid; filter; solid pure water twice; dry to obtain intermediate M3604mg, productivity 90%.

¹H-NMR (DMSO-d6, ppm): �� 2.87-2.89 (2H, m), 3.69-3.71 (2H, m), 4.72 (1H, s), 6.84 (1H, s), 6.89-6.91 (1H, d), and 7.74-7.75 (1H, d), 8.33 (1H, s), 8.55 (1H, s), 10.83 (1H, s) .ESI-MS:[M-H]^-=272.1.

(3) preparation of product

200.0mg (0.73mmol) M3 is dissolved in 10mLDMF, add the succinic anhydride of 146.5mg (1.46mmol) and the DMAP of catalytic amount, 50 DEG C of reactions are overnight, it is cooled to room temperature, concentration of reaction solution, column chromatography for separation obtains light yellow product 231mg, productivity 85%, and eluent is petroleum ether, ethyl acetate and methanol (10: 10: 1) mixed solvent.

¹H-NMR (DMSO-d6, ppm): �� 2.42 (4H, s), 3.04-3.08 (2H, m), and 4.30-4.34 (2H, m), 6.85 (1H, s), and 6.89-6.92 (1H, d), 7.73-7.76 (1H, d), 8.40 (1H, s), 8.57 (1H, s), 10.87 (1H, s), 12.15 (1H, s) .ESI-MS:[M-H]^-=372.1.

(2), the in vitro cytotoxic effect test of compound

Experimental technique: adopt MTT method that representative compound and the cisplatin medicine of the present invention have been carried out cytotoxic activity test. Taking the logarithm the cell counting of trophophase, be inoculated in 96 well culture plates, every hole is about 8000-10000 cell. Incubated overnight, is administered after cell attachment, sets administration group, positive controls and negative control group respectively. Compound to be measured is configured to storage liquid with D/W or the DMSO of 5%, becomes a series of concentration with cell culture medium before use, and wherein the final concentration of DMSO is less than 4 ��. Each concentration sets 3 multiple holes. Cultivating 72 hours after dosing, adding 20 �� l concentration is the MTT of 5mg/ml, hatches 4 hours, removes supernatant for 37 DEG C, and the DMSO adding 150 �� l dissolves first a ceremonial jade-ladle, used in libation. Under 490 wavelength, measure the OD value in every hole by microplate reader, and calculate suppression ratio, do concentration-suppression ratio curve and calculate IC₅₀Value.

Test example 1

Testing the compound 1-2 anti tumor activity in vitro to human breast cancer cell line Bcap-37, hepatocellular carcinoma H22, colon cancer cell HCT-116, ovarian cancer cell A2780, pancreatic cancer cell PANC-1, stomach cancer cell SGC-7901 and cisplatin resistance stomach cancer cell SGC-7901/CCCP, cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations, calculate suppression ratio and IC thereof₅₀Value evaluates the cytotoxic activity of medicine, and result is in Table 2.

The cytotoxic activity of table 2. compound 1 and 2

* .RF: drug-resistance factor=drug resistant cancer cells IC₅₀Value/normal cancerous cell IC₅₀Value.

* .nd: do not measure.

Test example 2

Testing the compound 3 anti tumor activity in vitro to human colon cancer cell HCT-116, stomach cancer cell SGC-7901 and cisplatin resistance stomach cancer cell SGC-7901/CCCP, cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations, calculate suppression ratio and IC thereof₅₀Value evaluates the cytotoxic activity of medicine, and result is in Table 3.

The cytotoxic activity of table 3. compound 3

* .RF: drug-resistance factor=drug resistant cancer cells IC₅₀Value/normal cancerous cell IC₅₀Value.

Test example 3

Testing the compound 4.6 anti tumor activity in vitro to human colon cancer cell HCT-116, lung cell A549, breast cancer cell MCF-7, hepatocellular carcinoma H22, stomach cancer cell SGC-7901 and cisplatin resistance stomach cancer cell SGC-7901/CCCP, cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations, calculate suppression ratio and IC thereof₅₀Value evaluates the cytotoxic activity of medicine, and result is in Table 4.

The cytotoxic activity of table 4. compound 4-6

* .RF: drug-resistance factor=drug resistant cancer cells IC₅₀Value/normal cancerous cell IC₅₀Value.

* .nd: do not measure.

Test example 4

Testing the compound 7 anti tumor activity in vitro to human glioma cell U87 and neurogliocytoma cell U251, cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations, calculate suppression ratio and IC thereof₅₀Value evaluates the cytotoxic activity of medicine, and result is in Table 5.

The cytotoxic activity of table 5. compound 7

Test example 5

Testing the compound 8 anti tumor activity in vitro to human breast cancer cell MDA-MB-231 and MCF-7, colon cancer cell HCT-116, stomach cancer cell SGC-7901 and cisplatin resistance stomach cancer cell SGC-7901/CCCP, cisplatin is as positive control. Observe compound suppression situation to growth of tumour cell under variable concentrations, calculate suppression ratio and IC thereof₅₀Value evaluates the cytotoxic activity of medicine, and result is in Table 6.

The cytotoxic activity of table 6. compound 8

* .RF: drug-resistance factor=drug resistant cancer cells IC₅₀Value/normal cancerous cell IC₅₀Value.

(3), the anti-tumor in vivo active testing of compound

Experimental technique: adopt nude mice, evaluates given the test agent and grows SGC-7901 cells nude mouse xenograft tumor with or without inhibitory action and action intensity.

Animal subject: BALB/c nude mouse, is provided by Ling Chang bio tech ltd, Shanghai. Laboratory animal production licence: SCXK (Shanghai) 2013-0018, the quality certification is numbered: 2013001812710, laboratory animal occupancy permit: SYXK (Soviet Union) 2011-0036. Age in days: 6w, body weight: 18-20g, sex: female, number of animals: often group 5, totally 35.

Medicine and reagent: cisplatin, compound 1 and CX-OH, be used that 5% glucose injection ultrasonic dissolution.

Group and dosage regimen are as follows:

Model control group: tail vein injection 5% G/W, 0.1ml/10g, once in a week, administration 3 times altogether;

Compound 1 (5mg/kg): tail vein injection 5mg/kg drug solution, 0.1ml/10g, once in a week, administration 3 times altogether;

Cisplatin (5mg/kg): the drug solution of tail vein injection 5mg/kg, 0.1ml/10g, once in a week, administration 3 times altogether;

CX-OH (5mg/kg): the drug solution of gavage 5mg/kg, 0.1ml/10g, once in a week, administration 3 times altogether;

CX-OH (5mg/kg): the drug solution of gavage 5mg/kg, 0.1ml/10g, every other day once, administration 11 times altogether;

Cisplatin+CX-OH (is 5mg/kg): tail vein injection cisplatin solution, gavage CX-OH solution, 0.1ml/10g, is weekly, altogether administration 3 times;

Cisplatin+CX-OH (is 5mg/kg): tail vein injection cisplatin solution, 0.1ml/10g, once in a week, administration 3 times altogether, gavage CX-OH, 0.1ml/10g, every other day once, administration 11 times altogether;

Test example 6

Collecting the SGC-7901gastriccarcinomacellline suspension cultivated, concentration is 1 �� 10⁷Individual/ml, is inoculated in axillary fossa on the right side of nude mouse with every 0.1ml subcutaneous. Transplanted tumor in nude mice vernier caliper measurement transplanted tumor diameter, after inoculating 11 days, tumor growth is to 100-150mm³Time by animal random packet, often group 5. Meanwhile, each group nude mice starts administration, and dosage regimen is shown in group and dosage regimen, uses the method measuring tumor footpath, dynamically observes the Graft Versus Tumor of given the test agent. After experiment terminates, putting to death nude mice immediately, operation strips tumor block and weighs.

The computing formula of gross tumor volume (tumorvolume, TV) is: TV=1/2ab², wherein a, b represent length and width respectively.

Result according to measuring calculates relative tumour volume (relativetumorvolume, RTV), and computing formula is:

RTV=V_t/V₀, wherein V₀(d during for sub-cage administration₀) measure gained gross tumor volume, V_tGross tumor volume during for measuring each time. The evaluation index of anti-tumor activity: Relative tumor rate of increase T/C (%), computing formula is as follows:

$T/C\left(\%\right)=\frac{T_{RTV}}{C_{RTV}}\×100$

Wherein, T_RTV: treatment group RTV; C_RTV: model group RTV.

The evaluation index of anti-tumor activity: inhibition rate of tumor growth (%), computing formula is as follows:

Average X �� SD represents, analyzes and carry out statistical procedures with t inspection between group, and application SPSS (StaffsticalPackagefortheSocialScience) 17.0 pairs of results carry out statistical analysis.

The inhibitory action that SGC-7901 cells nude mouse xenograft tumor is grown by table 7. given the test agent

Compare with model control group, * P < 0.05, * * P < 0.01.

Claims (4)

1. a class contains the tetravalence platinum complex of bio-active group, it is characterised in that described tetravalence platinum complex and platinum (IV) coordination compound, and its structure is such as shown in Formula II:

In Formula II, Y is OH or Cl; Bio represents bio-active group.

2. the preparation method that a class as claimed in claim 1 contains the tetravalence platinum complex of bio-active group, it is characterised in that described platinum (IV) coordination compound, the reaction equation shown in formula III carries out,

In formula III, Y is OH or Cl; Bio-OH represents has bioactive compound, and TBTU represents coupling reagent O-BTA-N, N, N ', N '-tetramethylurea Tetrafluoroboric acid, and TEA represents catalyst of triethylamine, and DMF represents solvent DMF, and DMSO represents solvent dimethyl sulfoxide;

Specifically adopt with the following method: equimolar reactant B io-OH and coupling reagent TBTU is mixed in dry DMF or DMSO, under stirring at room temperature, add equimolar TEA, it is subsequently adding equimolar Pt (IV) reactant A or B, reactant liquor stirs 12-48 hour under spending at 30-60 DEG C under nitrogen protection, then removal of solvent under reduced pressure, and concentrated solution separates through silica gel column chromatography, eluent is dichloromethane and methanol mixed solvent, obtains yellow solid product;

The structure such as Formulas I V of Pt (IV) reactant A and B represents,

Wherein A represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH)₂], reacted in water by cisplatin and hydrogen peroxide and prepare; B represents suitable, suitable, trans-[Pt (NH₃)₂Cl₂(OH) Cl], reacted in water by cisplatin and chlorosuccinimide and prepare.

3. the preparation method that a class according to claim 2 contains the tetravalence platinum complex of bio-active group, it is characterised in that described Bio-OH represents has bioactive compound structure shown in Formulas I (a-f),

Wherein, (a) is the anti-tumor drugs targeting silmitasertib of known entrance clinical trial, is a kind of Protein kinase C K2 inhibitor, referred to as CX-OH; B () is known marketed drug chlorambucil, be a kind of target spot be the antitumor drug of DNA, referred to as CHL-OH; C () is biotin, i.e. biotin, referred to as BT-OH; D () is the carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUA-OH; E () is the triazole carboxylic acid derivates of the natural product coumarin with antioxidation antitumaous effect, referred to as CUT-OH; F () is the carboxylic acid derivates of known antitumor drug SAHA, be a kind of PARP inhibitor, referred to as SAA-OH.

4. the preparation method that a class according to claim 2 contains the tetravalence platinum complex of bio-active group, it is characterised in that the preparation of described CUT-OH is undertaken by the reaction equation shown in Formula V,

Preparation CUT-OH adopts and is made by method:

1) sodium acetate of equimolar 2,4-4-dihydroxy benzaldehydes and acetoglycocoll and triplication is added in acetic anhydride, be stirred at reflux 4h, reactant liquor is cooled to room temperature, pours in frozen water, obtain yellow mercury oxide, filtering, wash twice with frozen water, drying solid obtains intermediate M1;

2) M1 being added concentrated hydrochloric acid: ethanol is in the mixed solution of 2: 1, reflux 1h, and reactant liquor is cooled to room temperature, pour in frozen water, then under condition of ice bath, it is slowly added to the sodium nitrite of qdx, after adding stirring 15min, it is slow added into the Hydrazoic acid,sodium salt of triplication, continue stirring 15min, filter out tan precipitate, washing, column chromatography for separation obtains intermediate M2, and eluent is petroleum ether: ethyl acetate is 5: 1 mixed solvents;

3) M2 is suspended in methanol, add equimolar fourth-3-alkynes-1-alcohol and a small amount of sodium ascorbate, 10min is stirred at room temperature under nitrogen protection, again the copper sulphate pentahydrate being dissolved in the qdx of water is joined in reactant liquor, room temperature lucifuge stirring 4h, then rotation is evaporated off the methanol in dereaction liquid, add water precipitation yellow solid, filter, gained solid pure water twice, dry to obtain intermediate M3;

4) M3 being dissolved in DMF, add the succinic anhydride of qdx and the DMAP of catalytic amount, 50 DEG C of reactions are overnight, it is cooled to room temperature, concentration of reaction solution, column chromatography for separation obtains light yellow product, and eluent is petroleum ether: ethyl acetate: methanol is 10: 10: 1 mixed solvents.