CN114075250B

CN114075250B - Preparation of platinum metal complex of oxafolium and application thereof in antitumor drugs

Info

Publication number: CN114075250B
Application number: CN202010840990.5A
Authority: CN
Inventors: 张跃茹; 向道凤; 谭相石; 颜道静
Original assignee: Hangzhou Xing'ao Biological Technology Co ltd
Current assignee: Hangzhou Xing'ao Biological Technology Co ltd
Priority date: 2020-08-20
Filing date: 2020-08-20
Publication date: 2023-01-13
Anticipated expiration: 2040-08-20
Also published as: WO2022036991A1; CN114075250A

Abstract

The invention belongs to the technical field of biological medicines, and discloses an innovative platinum metal complex which is formed by coordination of cyclohexanediamine, folic acid and metal platinum. Therefore, the innovative platinum coordination compound of the oxafolium has wide application prospect in preparing antitumor drugs.

Description

Preparation of platinum metal complex of oxafolium and application thereof in antitumor drugs

Technical Field

The invention belongs to the technical field of biological medicine, and particularly relates to a novel platinum metal complex of oxa-nervone [ Pt (DACH) (FA) ₂ ]Preparation, structural composition and application thereof in preparing antitumor drugs.

Background

Tumors are one of the major diseases seriously harming human life and health, and are manifested by abnormal cell hyperproliferation and differentiation. According to WHO statistics, annual cancer cases may increase to 2400 million worldwide. At the same time, cancer imposes a huge burden on the global economy. The following cancers are the main culprits of human death worldwide and mainly comprise: tracheal/bronchial cancer, lung cancer; liver cancer, gastric cancer, esophageal cancer, colorectal cancer, cancer of the reproductive system, prostate cancer, breast cancer, and cervical cancer. Chemotherapy, is a common clinical method used in cancer treatment at the present stage. Metal complex drugs such as platinum drugs belong to chemotherapeutic drugs and are widely used in the chemotherapy process. Clinically, mainly aiming at different cancer patients, effective platinum drugs are selected to inhibit the growth of tumor cells or promote the tumor cells to trigger a self-death mechanism, thereby achieving the aim of curing the cancer.

Metal complex drugs have been used clinically as early as the 70's in the last century. At present, there are first-line clinical anticancer drugs cisplatin and its derivative drugs, and also many metal complex drugs in clinical and preclinical research, including ruthenium, gold, iridium, etc. The platinum antineoplastic drugs become one of the most effective drugs for clinically treating malignant tumors at present due to the unique action mechanism and wide anticancer spectrum. However, the problems of large toxic and side effects, unclear drug resistance or cross drug resistance, targeting property and the like become obstacles for limiting the expansion of clinical application. According to statistics, 70-80% of the current chemotherapy or combined chemotherapy treatment schemes for malignant tumors use platinum drugs. Following cisplatin, 6 platinum-based antitumor drugs such as carboplatin (carboplatin), nedaplatin (nedaplatin), oxaliplatin (oxaliplatin), sulbactin (SKI-2053R), lobaplatin (lobaplatin) and miriplatin (miriplatin) are listed in each country in succession. Although platinum antineoplastic drugs have achieved great success, there are many obstacles to clinical application, such as no targeting, easy generation of toxic and side effects, drug resistance or cross-drug resistance. Cisplatin has special effect on testicular cancer, and the cure rate is improved from 10% to 85%. The cis-platinum drugs have many side effects, including: renal toxicity; nausea, vomiting; neurotoxicity; ototoxicity; alopecia; electrolyte disturbance. Clinically, the composition can also be used together with bleomycin (bleomycin), paclitaxel (paclitaxel) and 5-fluorouracil (5-fluorouracil) to achieve better treatment effect. Nevertheless, the use of platinum-based anticancer drugs and the like is limited by their toxic side effects and the inherent and acquired resistance of tumor cells. In the clinical treatment process, the problem that people need to solve is that the metal complex medicines with higher efficiency and lower toxicity are required. Therefore, based on the monofunctional cisplatin metal complex, the cisplatin metal complex has targeting and composite functions, can increase efficiency and reduce toxicity, and the research of the innovative anticancer platinum metal complex becomes an international hotspot.

Folic acid is an important B-group water-soluble vitamin, also called vitamin M, which was isolated from spinach in 1941 and named because of its abundant content in green leaves, also called pteroylglutamic acid. As the importance of folic acid in diet is gradually recognized, especially the research on folic acid and birth defects, cardiovascular diseases and tumors is deepened, folic acid becomes an extremely important micronutrient. Folic acid plays an important role in purine and pyrimidine synthesis, amino acid interconversion, and certain methylation reactions. The formyl group as carried by tetrahydrofolic acid is used in the following various reactions: (1) carbons at positions 2, 5, and 8 in purine synthesis; (2) glycine to serine; (3) homocysteine is methylated to form methionine; (4) synthesizing thymine from uracil; (5) synthesizing choline from ethanolamine; (6) methylation of nicotinamide to form N' -methylnicotinamide, etc. The lack of folic acid in humans produces megaloblastic anemia, glossitis and diarrhea.

The research preferably prepares the innovative platinum coordination compound [ Pt (DACH) (FA) of Osa leaf by using folic acid as the coordination molecule of metal platinum and the coordination reaction of folic acid and diamine ligand with transition metal such as platinum and the like ₂ ]. Research shows that the preferred olsalazine coordination compound has good water solubility, obvious curative effect of inhibiting solid tumor of mice and obviously reduced toxic and side effect compared with oxaliplatin. Therefore, the innovative platinum metal complex of oxafolium oxanthi sibirica has good application prospect in preparing antitumor drugs.

Disclosure of Invention

The invention provides a composition of a novel folic acid platinum metal complex, a preparation method and application thereof in antitumor drugs. The platinum metal complex is formed by coordination of cyclohexanediamine and folic acid with metal platinum. The innovative platinum metal complex has good water solubility. The compound has better anti-tumor activity than oxaliplatin in a mouse solid tumor model test, has better safety, and obviously reduces toxic and side effects than cisplatin and oxaliplatin.

Detailed Description

The present invention will be described in detail with reference to examples. In the present invention, the following examples are given to better illustrate the present invention and are not intended to limit the scope of the present invention.

Example 1: novel platinum metal complex of oxafolin [ Pt (DACH) (FA) ₂ ]Preparation and characterization of all reagents were analytical grade, purchased from Sigma-Aldrich. [ Pt (DACH) (FA) ₂ ]The specific synthesis steps are as follows: dissolving potassium tetrachloroplatinate (1 mmol) and potassium iodide (8 mmol) in 20 ml of purified water under stirring, adding cyclohexanediamine (DACH, 2 mmol) dropwise, reacting under stirring for 1 hr to give yellow precipitate, filtering, washing with water to give yellow powder, and drying to obtain intermediate [ Pt (D)ACH)(I) ₂ ]. Dissolving the intermediate yellow powder in 50 ml water, stirring for overnight reaction, filtering to remove silver iodide precipitate, adding dropwise sodium hydroxide solution (2 mmol) of folic acid into the filtrate, heating for 2 days, filtering, separating and purifying the filtrate with ion exchange column, concentrating, and drying to obtain final product [ Pt (DACH) (FA) ₂ ]813. Mg, yield 68%.

The final product, brown yellow microcrystalline powder, was verified by mass spectrometry, elemental analysis, and platinum metal content analysis to be [ Pt (DACH) (FA) ₂ ]. The peak of the highest nuclear to cytoplasmic ratio was 1190 by mass spectrometry, corresponding to the target product molecular weight of 1189. Elemental analysis nuclear platinum metal analysis results were as follows:

TABLE 1 [ Pt (DACH) (FA) ₂ ]Elemental analysis and analysis results of platinum Metal content

Example 2: detection Using mouse tumor bearing model [ Pt (DACH) (FA) ₂ ]The anti-tumor effect of the compound is the inhibition effect on the growth of subcutaneous transplantation tumor of the mouse.

Animal(s) production

BALB/C normal mice, male, 20-22g in weight, 7-8 weeks old, SPF grade, purchased from shanghai sierk laboratory animals llc [ laboratory animal quality certification no: SCXK (Shanghai) 2007-0005].

Feeding conditions

All mice were left free to feed and drink water and were kept at room temperature (25 + -2) ° c. The feed and water are sterilized by high pressure, and all experimental feeding processes are SPF grade.

Dose setting

Mice were given by intraperitoneal injection, [ Pt (DACH) (FA) ₂ ]1 dose group 15mg/kg (platinum content: 2.4 mg/kg) was set.

Test control

Negative control: physiological saline solution

Positive control: oxaliplatin in a dose of 5mg/kg (platinum content: 2.4 mg/kg);

method of administration

The administration route is as follows: abdominal injection

Administration volume: 100 microliter/piece; the administration times are as follows: the administration was continued for 27 days, 1 time every other day.

Number of animals per group: 10 are

The mouse colorectal cancer cell line CT26 and the mouse breast cancer cell line 4T1 are purchased from cell banks of Chinese academy of sciences.

The main steps of the test

Establishment and intervention of tumor model mouse

Cell culture, passage, cell collection at log phase, concentration of 1.0X 10 per ml ⁷ ) Respectively preparing mouse colorectal cancer cell line CT26, transplanting the mouse colorectal cancer cell line CT26 to a BalB/C common mouse and a mouse breast cancer cell line 4T1, transplanting the mouse colorectal cancer cell line CT to a BalB/C common mouse, and observing the anti-tumor effect of different drugs. Mice right forelimb axillary injection of 0.2ml cell suspension (cell number 2.0X 10) ⁶ One/one), the tumorigenicity is successful about 8 days. According to the experimental categories of two mouse tumor models, the two groups are respectively divided into 3 groups randomly, including a model control group, an oxaliplatin positive drug control group and an innovative drug oxafolin [ Pt (DACH) (FA) ₂ ]And (4) administration groups. The administration was 1 time every other day for 27 consecutive days. After 27 days, the mice were sacrificed and tumor body weights were taken and tumor inhibition rates were calculated.

Statistical analysis

Data are expressed in x ± s, processed using SPSS10.0 software, and the significance of tumor weight differences for each group was compared using a one-way ANOVA test, with significance level a =0.05.

Results of the experiment

A new type of innovative drug [ Pt (DACH) (FA) ] was successfully prepared after subcutaneous inoculation of tumor cells in mice ₂ ]Can obviously inhibit the growth of tumor, and the tumor weight is obviously lower than that of a negative control model group (P) after the administration for 27 days<0.05， P<0.01 The medicine effect of inhibiting tumor growth is better than that of oxaliplatin), the survival rate of mice is obviously improved, and the toxic and side effect of the medicine is obviously reduced compared with that of the oxaliplatin. Specific results are shown in the following table.

TABLE 2 [ Pt (DACH) (FA) ₂ ]For BalB/C mouseEffect of colorectal cancer cells CT26 subcutaneous transplantation tumor (n =10, mean. + -. SD)

Note: * P <0.05vs negative control; * P <0.01vs negative control group.

TABLE 3 [ Pt (DACH) (FA) ₂ ]Inhibition of BalB/C murine breast cancer 4T1 subcutaneous transplantable tumor (n =10, mean ± SD)

Note: * P <0.05vs negative control; * P <0.01vs negative control group.

Claims

1. The platinum metal complex is characterized in that the chemical formula of the platinum metal complex is [ Pt (DACH) (FA) ₂ ]Wherein DACH is cyclohexanediamine and FA is folate.

2. A process for preparing the platinum metal complex of oxaneri of claim 1, comprising: taking potassium tetrachloroplatinate as a platinum metal raw material, firstly reacting with KI to generate potassium tetraiodoplatinate; the potassium tetraiodoplatinate reacts with cyclohexanediamine to generate [ Pt (DACH) (I) ₂ ]Complex intermediate, [ Pt (DACH) (I) ₂ ]Then reacting with folic acid in the presence of silver nitrate under heating and stirring, separating and purifying the filtrate to obtain platinum complex [ Pt (DACH) (FA) ₂ ]Liquid, concentration and drying to obtain a powder solid sample.

3. Use of the platinum olsa-leaf complex according to claim 1 for the preparation of an antitumor agent for the treatment of various types of tumors including colorectal, breast, testicular, ovarian, prostate, lung, nasopharyngeal, esophageal, malignant lymphoma, head and neck, thyroid, osteogenic sarcoma, bladder, cervical and germ cell tumors.

4. The use of claim 3, wherein the anti-tumor drug comprises unit preparations with different specifications and pharmaceutically acceptable drug preparations, and the administration mode of the anti-tumor drug comprises one or more of intravenous injection, intramuscular injection, subcutaneous injection, intravenous drip, nasal drip and oral administration.