8-hydroxy dictamnine platinum complex with high activity in vitro and vivo as well as synthesis method and application of 8-hydroxy dictamnine platinum complex

Abstract

The invention discloses an 8-hydroxyl dictamnine platinum complex with high activity in vitro and vivo as well as a synthesis method and application thereof. According to the invention, 8-hydroxy dictamnine is used as an active ligand and reacts with cis-Pt (DMSO) 2Cl2 platinum salt to synthesize the 8-hydroxy dictamnine platinum complex. The 8-hydroxyl dictamnine platinum complex can inhibit the growth of human ovarian adenocarcinoma cis-platinum drug-resistant cells SK-OV-3 / DDP in a targeted manner, shows excellent anti-tumor activity, has potential medicinal value, and is expected to be used for preparing various anti-tumor drugs.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to an 8-hydroxybactamine platinum complex with high in vitro and in vivo activity, a synthesis method and application thereof. Background Art

[0002] The nephrotoxicity and toxicity of cisplatin-based drugs limit their clinical application, motivating scientists to develop a series of novel platinum-based anticancer drugs that can target and overcome drug resistance. However, the synthesis and anticancer activity of 8-hydroxybactam platinum complexes have not been reported in the literature. Summary of the Invention

[0003] One of the purposes of the present invention is to provide an 8-hydroxybactamine platinum complex.

[0004] Specifically, the 8-hydroxybactam platinum complex has a chemical formula of [Pt(Rob)Cl2](RoPt), and its chemical structure is shown below:

[0005]

[0006] A second object of the present invention is to provide a method for synthesizing the above-mentioned 8-hydroxybactamine platinum complex.

[0007] Specifically, the synthesis method of the above-mentioned 8-hydroxyrodecanoic acid platinum complex comprises weighing 8-hydroxyrodecanoic acid (H-Rob) and cis-Pt(DMSO)2Cl2 platinum salt, adding a solvent, reacting at 45°C for 2 days and at 68°C for 1 day under sealed conditions, cooling, and filtering to obtain the 8-hydroxyrodecanoic acid platinum complex.

[0008] Its synthetic route is as follows:

[0009]

[0010] The solvent is methanol (CH3OH) and dimethyl sulfoxide (DMSO), or methanol (CH3OH) and acetone.

[0011] Another object of the present invention is to provide the application of the above-mentioned 8-hydroxybactamine platinum complex.

[0012] Specifically, the present invention relates to the use of the above-mentioned 8-hydroxybuctamidine platinum complex in the preparation of anti-tumor drugs and the use of the above-mentioned 8-hydroxybuctamidine platinum complex in the preparation of drugs for the targeted treatment of ovarian cancer.

[0013] Compared with the existing technology, the present invention uses 8-hydroxyrobin (H-Rob) as the active ligand to react with cis-Pt(DMSO)2Cl2 platinum salt to synthesize 8-hydroxyrobin platinum complex RoPt, and investigates its anticancer activity and toxicity against human ovarian adenocarcinoma SK-OV-3 cells, human ovarian adenocarcinoma cisplatin-resistant SK-OV-3 / DDP cells, and normal liver HL-7702 cells. The experimental results show that the complex RoPt has the best anticancer effect on SK-OV-3 / DDP cancer cells, with an IC of 1. 50 The value was 6.42±0.21 μM, demonstrating significantly greater activity than H-Rob, cis-Pt(DMSO)2Cl2, and cisplatin. Furthermore, it exhibited minimal toxicity to normal HL-7702 cells, demonstrating that the complex RoPt could target and inhibit the proliferation of SK-OV-3 / DDP cancer cells. In vivo tumor inhibition experiments demonstrated that the complex exhibited a potent tumor inhibition effect in nude mice bearing SK-OV-3 ovarian adenocarcinoma, with an inhibition rate of up to 37.1%. In summary, the 8-hydroxybactam platinum complex RoPt exhibited superior in vitro and in vivo antitumor activity, possessing potential medicinal applications and promising potential for the preparation of various antitumor drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is the X-ray single crystal diffraction pattern of the 8-hydroxybactam platinum complex RoPt prepared in Example 1 of the present invention;

[0015] Figure 2 This is a hydrogen nuclear magnetic resonance spectrum of the 8-hydroxybactam platinum complex RoPt prepared in Example 1 of the present invention;

[0016] Figure 3 This is an infrared spectrum of the 8-hydroxybactam platinum complex RoPt prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0017] The present invention will be further described below with reference to specific examples, but the present invention is not limited to these examples.

[0018] Example 1

[0019] In a 15.0 cm thick-walled drug-resistant tube, 0.1 mmol of 8-hydroxyrobin (H-Rob) and cis-Pt(DMSO)2Cl2 platinum salt were weighed, respectively, followed by the addition of 1.5 mL of CH3OH and 0.1 mL of DMSO. After the drug-resistant bottle was capped, the reaction was incubated at 45°C for 2 days and 68°C for 1 day, then cooled to 37°C, the bottle cap was opened, and the compound RoPt was obtained with a yield of 86.4%.

[0020] The obtained product RoPt was identified:

[0021] (1) X-ray diffraction pattern of compound RoPt, the spectrum is as follows Figure 1 shown.

[0022] (2) The H NMR spectrum of compound RoPt is as follows Figure 2 shown.

[0023] 1 H NMR (600MHz, DMSO-d6) δ8.17(d,J=2.7Hz,1H),7.66(d,J=2.7Hz,1H),7.32(dd,J =11.9,5.1Hz,2H),6.96(dd,J=6.8,2.0Hz,1H),4.56(s,3H),2.50(overlap,6H).

[0024] (3) Elemental analysis results are shown in Table 1.

[0025] Table 1 Elemental analysis results of compound RoPt in the embodiment

[0026]

[0027]

[0028] (4) Infrared spectrum data of compound RoPt.

[0029] IR(KBr):3437,3152,3121,3083,3013,2999,2952,2916,1929,1750,1616,1585,1557,1520,1468,1441,1416,1404,1368,1342,1317 ,1299,1290,1242,1225,1198,1160,1134,1103,1062,1029,991,944,922,885,864,813,805,742,722,691,641,586,578,555,527cm -1 .

[0030] Thus, the obtained product RoPt can be determined, and its structural formula is as follows:

[0031]

[0032] Example 2

[0033] In a 15.0 cm thick-walled drug-resistant tube, 0.1 mmol of 8-hydroxyrobin (H-Rob) and cis-Pt(DMSO)2Cl2 platinum salt were weighed, respectively, followed by the addition of 1.5 mL of CH3OH and 0.1 mL of acetone. After the lid of the drug-resistant bottle was closed, the reaction was incubated at 45°C for 2 days and 68°C for 1 day, then cooled to 37°C, the bottle cap was opened, and the compound RoPt was obtained with a yield of 80.9%.

[0034] In order to fully illustrate the use of the 8-hydroxybactamine platinum complex RoPt of the present invention in pharmaceutical manufacturing, the applicant conducted in vitro and in vivo anti-tumor activity experiments on it.

[0035] 1. Experimental study on the proliferation inhibition activity of 8-hydroxybactam platinum complex RoPt on three human cell lines

[0036] 1. Cell lines and cell culture

[0037] In this study, three human cell lines were selected: human ovarian adenocarcinoma SK-OV-3 cells, human ovarian adenocarcinoma cisplatin-resistant SK-OV-3 / DDP cells, and normal liver HL-7702 cells.

[0038] All human cell lines were cultured in RPMI-1640 medium containing 100 U / mL penicillin, 10 wt% calf blood, and 100 U / mL streptomycin in an incubator at 37° C. with a volume concentration of 5% CO 2 .

[0039] 2. Preparation of Test Compounds

[0040] The purity of all compounds used must be ≥95%. Their DMSO stock solutions are diluted with physiological buffer to a final solution of 20 μmol / L (final DMSO concentration ≤1%), and the degree of inhibition of each compound on the growth of normal cells or selected tumor cells at this concentration is tested.

[0041] 3. Cell Growth Inhibition Experiment

[0042] The specific steps are as follows:

[0043] (1) Cells were digested, counted, and prepared to a concentration of 5×10 4 100 μL of cell suspension was added to each well of a 96-well cell culture plate and cultured in a 37°C, 5.0% CO2 incubator for 24.0 h.

[0044] (2) Dilute the drug to the desired concentration using RPMI-1640 medium, and add 100 μL of the corresponding drug-containing medium to each well of a 96-well plate to achieve final concentrations of 0, 1.0, 2.5, 5.0, 10.0, 20.0, and 50.0 μM, respectively. Set up six parallel experiments for each concentration. After adding the drug, continue culturing in a 37°C, 5% CO2 incubator for 48 h.

[0045] (3) Perform CCK-8 staining and measure the OD value at a wavelength of λ = 450nm: add 10μL CCK-8 to each well, continue to culture in the incubator for 2h, shake on a shaker for 10min to gently mix, and then measure the OD value of each well with a microplate reader at a wavelength of λ = 450nm. Calculate the inhibition rate of each compound on the growth of the selected cell line, and then calculate the IC value of each test compound on the selected cell line using the Bliss method. 50 The results are shown in Table 2 below.

[0046] Table 2. IC values ​​of compound RoPt against various cell lines 50 Value (μM)

[0047]

[0048] From IC 50 The results of activity screening showed that the complex RoPt had the best anticancer effect on SK-OV-3 / DDP cancer cells, with an IC 50 The value is 6.42±0.21μM, and its activity is much greater than that of H-Rob, cis-Pt(DMSO)2Cl2 and cisplatin; and it has low toxicity to normal HL-7702 cells; indicating that the complex RoPt can target and inhibit the proliferation of SK-OV-3 / DDP cancer cells.

[0049] 2. In vivo tumor inhibition experiment in tumor-bearing nude mice

[0050] Human ovarian adenocarcinoma SK-OV-3 cells in the logarithmic growth phase were collected and diluted to 5.0×10 7 Use a 1.0mL syringe to draw 0.2mL of suspension, which contains approximately 1×10 7 Then, the cells were inoculated into the subcutaneous tissue of the right armpit of nude mice. 3 Human breast squamous cell carcinoma HCC1806 cells were passaged in nude mice for 4 generations. After their growth stabilized, mice with vigorous tumor growth and no ulceration were selected and sacrificed by cervical dislocation. The animal skin was disinfected with 75% medical alcohol, and the tissue blocks were dissected to remove the necrotic part. The tumor tissue was cut into 1.5mm pieces. 3The left and right small pieces were inoculated into the subcutaneous tissue of the right axilla of nude mice using a trocar. The diameter of the transplanted tumor was measured with an electronic vernier caliper. When the tumor volume grew to 100-300 mm 3 The animals were randomly divided into groups.

[0051] Nude mice bearing human ovarian adenocarcinoma SK-OV-3 cells were randomly divided into a vehicle group, a drug-addition group, and a cisplatin-positive control group, with seven animals per group. Intraperitoneal injection of the compound was initiated on the day of grouping, with the compound administered every two days and cisplatin administered every other day. Tumor diameter and body weight were measured every three days. Mice were sacrificed by cervical dislocation on day 21, and tumors were dissected, weighed, photographed, and the tumor inhibition rate calculated.

[0052] Tumor volume calculation formula: V = a × b 2 / 2, where a is the major diameter and b is the minor diameter;

[0053] Relative tumor volume RTV = V t / V0, where V t is the volume at each measurement, and V0 is the volume at the time of grouping;

[0054] Relative tumor growth rate T / C% = (T RTV / C RTV )×100%;

[0055] Tumor growth inhibition rate (%) = (average tumor weight of vehicle group - average tumor weight of treatment group) / average tumor weight of vehicle group × 100%.

[0056] Table 3. In vivo tumor inhibition results of the complex RoPt on nude mice bearing human ovarian adenocarcinoma SK-OV-3 tumors

[0057]

[0058] In vivo tumor inhibition experiments, as shown in Table 3, demonstrate that the complex RoPt exhibits a moderate tumor inhibition effect in a nude mouse model bearing human ovarian adenocarcinoma SK-OV-3, with an inhibition rate of 37.1%. In summary, the complex YRoPt exhibits superior antitumor activity in vitro and in vivo, possesses potential medicinal value, and is expected to be used in the preparation of various antitumor drugs.

[0059] In summary, the 8-hydroxybactam platinum complex RoPt of the present invention exhibits excellent in vitro and in vivo antitumor activity and selectivity, as well as good cytotoxicity selectivity and overcoming cisplatin resistance. It has good potential medicinal value and is expected to be used in the preparation of various antitumor drugs.

Claims

1. A highly active 8-hydroxydandrogen platinum complex in vivo and in vitro, characterized in that: The chemical structure is shown below:

2. The method for synthesizing the 8-hydroxydandrogenine platinum complex according to claim 1, characterized in that: Take 8-hydroxyrodecanoic acid (H-Rob) and cis-Pt(DMSO)2Cl2 platinum salt, then add solvent, react at 45°C for 2 days under sealed conditions, then react at 68°C for 1 day, cool to 37°C, filter, and obtain 8-hydroxyrodecanoic acid platinum complex.

3. The method for synthesizing 8-hydroxydandrogenine platinum complex according to claim 2, characterized in that: The solvent is methanol and dimethyl sulfoxide, or methanol and acetone.

4. The method for synthesizing 8-hydroxydandrogenine platinum complex according to claim 3, characterized in that: The solvent is 1.5 mL of methanol and 0.1 mL of dimethyl sulfoxide.

5. The method for synthesizing 8-hydroxydandrogenine platinum complex according to claim 3, characterized in that: The solvent was 1.5 mL of methanol and 0.1 mL of acetone.

6. Use of the 8-hydroxydandrogenine platinum complex obtained according to claim 1 in the preparation of anti-tumor drugs.

7. Use of the 8-hydroxydandrogenine platinum complex obtained according to claim 1 in the preparation of drugs for targeted treatment of ovarian cancer.

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