CN110724127A

CN110724127A - Synthetic method and anti-tumor application of seleno-axitinib

Info

Publication number: CN110724127A
Application number: CN201911196493.XA
Authority: CN
Inventors: 郁彭; 萨旭仁贵; 张佳伟; 崔钰慧; 赵连波; 郝磊; 滕玉鸥; 王栋
Original assignee: Tianjin University of Science and Technology
Current assignee: Tianjin University of Science and Technology
Priority date: 2019-11-29
Filing date: 2019-11-29
Publication date: 2020-01-24
Anticipated expiration: 2039-11-29
Also published as: CN110724127B

Abstract

The invention relates to a synthetic method of a seleno-axitinib drug and application of the seleno-axitinib drug in antitumor drugs, wherein the synthetic method comprises the following steps: reacting compound 4 to form selenoacetinib. In vitro cell experiments prove that compared with the axitinib, the seleno-axitinib has obvious curative effect on treating the kidney cancer, obviously reduces the side effects of the axitinib such as blood pressure rise, weight reduction, liver enzyme, blood creatinine rise and the like, and provides a better alternative scheme for the optimization and clinical application of the antitumor drug axitinib.

Description

Synthetic method and anti-tumor application of seleno-axitinib

Technical Field

The invention belongs to the field of organic synthesis and the field of medicine application, and relates to a seleno-axitinib compound, which comprises synthesis, evaluation and application.

Background

Targeted treatment of cancer is currently an important therapeutic approach for controlling patients in the middle and late stages. Greatly prolongs the life cycle of the patient and obviously improves the life quality.

Axitinib is a targeted VEGFR1, 2, 3 inhibitor that was approved by the U.S. Food and Drug Administration (FDA) in 2012 for the treatment of renal cell carcinoma for which other systems have failed therapy, and is second-line drug administration. The axitinib inhibits the neogenesis and regeneration of tumor vessels by blocking VEGFR 2. Meanwhile, the use of the axitinib is often accompanied by the occurrence of hypertension and other side effects.

Selenium is a trace element indispensable to the life activities of the organism, and has wide biological effects, including antioxidation, induction of tumor cell apoptosis, blood pressure regulation, organism immunity enhancement and the like. Organic selenium such as selenocysteine and selenomethionine can remove oxygen free radical, protect cardiovascular system, and inhibit tumor cell growth and tumor blood vessel formation.

In view of the urgent need of developing novel targeted anticancer drugs, the structure of the existing targeted drugs on the market is needed to be optimized in the field, and the invention utilizes the principle of biological electron isostere to replace the sulfur element of the compound axitinib with the selenium element for biological evaluation to obtain the anticancer drug seleno-axitinib with small side effect and good curative effect.

Disclosure of Invention

The invention provides a synthetic method of seleno-axitinib and application of the seleno-axitinib in preparing antitumor drugs.

The purpose of the invention is realized by the following technical scheme:

the structural formula of the seleno-axitinib is as follows:

the seleno-axitinib compound is obtained by the following synthetic route:

the compounds of the present invention may be administered by a variety of routes, such as orally, intraarterially, subcutaneously, intramuscularly, intraluminally, peritumorally, and the like. The administration route and the administration dose may vary depending on the subject of administration, the administration route or the formulation form of the drug, but it is a prerequisite to ensure that the composition can achieve an effective blood concentration in the mammal.

The invention has the advantages and positive effects that:

1. the compound is a novel anti-tumor drug, has a remarkable curative effect on the treatment of kidney cancer, and remarkably reduces the side effects of increasing blood pressure, reducing weight and the like compared with the original drug of axitinib.

2. The compound is a high-activity antitumor drug composition, and in-vivo experiments are adopted for research to find that the composition keeps the antitumor effect of the axitinib, and obviously reduces the liver and kidney damage level caused by the axitinib compared with the original drug of the axitinib.

Drawings

FIG. 1 Effect of Axitinib and Selenoaxitinib on vascular permeability of mouse skin

FIG. 2 Effect of Axitinib and Selenoaxitinib on the systolic blood pressure of Wistar rats

FIG. 3 Effect of Axitinib and Selenoaxitinib on VEGFR-2 levels in mouse sera

FIG. 4 Effect of Asatinib and Selenoacetinib on liver enzyme (ALT)

FIG. 5 Effect of Asitinib and Selenoacetatinib on serum creatinine levels

FIG. 6 is a nuclear magnetic hydrogen spectrum of Selenacitinib in deuterated dimethyl sulfoxide;

figure 7 is a nuclear magnetic carbon spectrum of seleno-axitinib in deuterated dimethyl sulfoxide;

Detailed Description

The following detailed description of the embodiments of the present invention is provided for the purpose of illustration and not limitation, and should not be construed as limiting the scope of the invention.

The raw materials used in the invention are conventional commercial products unless otherwise specified; the methods used in the present invention are conventional in the art unless otherwise specified.

The structural formula of the seleno-axitinib compound is shown as the following figure:

the synthetic route of the seleno-axitinib compound is shown as the following figure:

the invention particularly comprises a synthetic method of the seleno-axitinib, which comprises the following steps:

compound 1(1.0g, 2.32mmol), Compound 2(1.99g, 4.64mmol), copper sulfide (4.43mg, 46.37. mu. mol), iron powder (77.69mg, 1.39mmol), potassium carbonate (320.45mg, 2.32mmol) and dimethyl sulfoxide (10mL) were added to a two-necked round bottom flask, the reaction was replaced three times with argon, the solution was warmed to 140 ℃ under argon and stirred, and the reaction was followed by TLC until the reaction was complete. Extraction was performed with ethyl acetate (3 × 100mL), the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and purified by using a developing solvent petroleum ether/ethyl acetate (10: 1) and 200-mesh 300-mesh silica gel column chromatography to obtain 3912 mg of the compound with a yield of 76%.

Compound 3(0.5g, 0.96mmol) was added to a solution consisting of methanol (5mL) and water (0.5mL), followed by p-toluenesulfonic acid monohydrate (1.1g, 5.79mmol), the solution was warmed to 70 ℃ and stirred, and the reaction was followed by TLC until the reaction was complete. After the reaction, the reaction solution was cooled to room temperature, 1mL of ice water was added thereto, the mixture was stirred for 20 minutes, the reaction solution was filtered, and the filter cake was dried in the dark to obtain 40.41g of the compound with a yield of 94%.

Compound 4(200mg, 0.46mmol) was added to 4mL of methylamine alcohol solution, and the reaction was stirred under a closed condition at 70 ℃ for 36 hours. After the reaction is finished, the solvent is evaporated to dryness under reduced pressure, 2mL of methyl tert-butyl ether is added for pulping, filtering and drying to obtain 180mg of seleno-axitinib, white solid and the yield is 90%.¹H NMR(400MHz，DMSO-d₆)δ13.43(s，1H)，8.61-8.58(m，2H)，8.24(d，J＝8.4Hz，1H)，7.97(d，J＝16.4Hz，1H)，7.86(s，1H)，7.84-7.80(m，1H)，7.69-7.67(m，2H)，7.60(d，J＝16.4Hz，1H)，7.37(d，J＝8.0Hz，1H)，7.30-7.22(m，3H)，6.90-6.87(m，1H)，2.81(d，J＝4.0Hz，3H).¹³C NMR(100MHz，DMSO-d₆)δ167.9，154.9，149.6，142.1，136.9，135.6，133.5，131.0，129.5，129.3，128.7，127.9，125.4，123.7，122.7，122.5，121.9，120.7，118.5，26.3.

Example 1

Inhibition effect of seleno-axitinib on growth of kidney cancer in Balb/c nude mouse subcutaneous xenograft tumor model

The suspension of the axitinib and the seleno-axitinib in 0.5 percent sodium carboxymethylcellulose is administered by intragastric administration for 15 days. The concentration of the low-dose 30mg/kg drug and the concentration of the high-dose 60mg/kg drug are respectively set for the axitinib and the seleno-axitinib. The results are shown in Table 1.

TABLE 1 inhibition of renal cancer growth by Axitinib and losartan monomers and combinations

From the above results, it can be seen that axitinib has a significant inhibitory effect on the growth of kidney cancer and is dose-dependent, and that seleno-axitinib also has a significant inhibitory effect on kidney cancer and is dose-dependent.

Example 2

Effect of Asitinib and Selenoacetatinib on vascular permeability of skin (Miles test)

A C57BL/6N mouse was gavaged with a single dose of Afatinib (10mg/kg) and seleno-Afatinib (10mg/kg), the tail vein was injected with Evans blue dye, the abdominal area was shaved after anesthesia, PBS and 1% VEGFA were injected into each side of the abdominal area, and after 4 hours, the abdominal area was vertically cut by about 3-4 cm from the lower abdomen to the chest with scissors. The dye aggregation zone was removed. After the sample is dried, formamide is added, evans blue dye is extracted, and the absorbance value is detected.

The results are shown in fig. 1, and it can be seen from the control group that VEGFA significantly increases vascular permeability. This effect is blocked by the VEGFR inhibitor axitinib, which reduces VEGFA-induced high vascular permeability, and is one of its mechanisms to inhibit tumor growth. The seleno-axitinib also achieves the same effect of inhibiting vascular permeability.

Example 3

Effect of Axitinib and Selenoaxitinib on blood pressure of Wistar rat

8 Wistar rats of 7 weeks of age were randomly selected, 2 rats per group, and individually gavage Axitinib (25mg/kg), Selenoaxitinib (25mg/kg) and control sodium carboxymethylcellulose were administered twice daily for 7 consecutive days. Systolic blood pressure changes were measured daily.

The systolic blood pressure test results on day 7 are shown in figure 2, compared with the control group, the systolic blood pressure of the rat is obviously increased by the axitinib by about 20%, and the blood pressure increase level of the seleno-axitinib is lower than that of the axitinib.

Example 4

Effect of Axitinib and Selenoaxitinib on body weight of Wistar rats

The use of the axitinib can cause the weight loss, the experiment uses the continuous administration of the axitinib and the seleno-axitinib with different concentrations for 7 days to detect the influence of various groups of compounds on the weight of rats, and the experimental result is shown in table 2.

TABLE 2 Effect of Axitinib and Selenoaxitinib on Wistar rat body weight

From the above results, it can be seen that the compound administration for 7 days increased the body weight of the control group by 20.1g, the axitinib group by only 3.1g, and the selenoacetitinib by 34.6 g. It can be concluded that axitinib affects the increase in body weight, whereas seleno-axitinib has a positive effect on body weight.

Example 5

Effect of Axitinib and Selenoaxitinib on serum VEGFR-2 levels

VEGFR-2 levels in blood serum of Balb/c nude mice after administration of different concentrations of Asitinib and seleno-Asitinib, and inhibition effect of Asitinib and seleno-Asitinib on VEGFR-2 was evaluated

The results are shown in figure 3, and the concentration of the obtained product is dependent on that of Astinib which inhibits VEGFR-2 in serum. Selenoacetinib has the same effect and is concentration-dependent.

Example 6

The effect of axitinib and selenoacetitinib on liver enzyme (ALT).

Glutamate-pyruvate transaminase (ALT) is a commonly used liver function detection index and can reflect the functional state of the liver to a certain extent. In this experiment, the ALT level in rat serum was used to evaluate the effect of axitinib and selenoacetitinib on liver function.

The results are shown in figure 4, the anticancer drug axitinib significantly increased ALT levels in serum, while the increase in ALT levels in serum of seleno-axitinib was much lower than that of axitinib.

Example 7

The effect of axitinib and selenoacetinib on blood creatinine (Cr).

Serum creatinine is an important indicator for the assessment of renal function, and an increase in serum creatinine level is often directly associated with a decrease in the rate of creatinine clearance in the kidney. In this experiment, the effect of axitinib and selenoacetitinib on renal function was evaluated by the creatinine level in the serum of Balb/c nude mice.

The results are shown in figure 5, and the blood creatinine level of the mice is remarkably increased by the Asitinib, while the serum creatinine level of the seleno-Asitinib tends to be normal.

Although the embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the embodiments disclosed.

Claims

1. An anti-tumor compound characterized by: the compound is selenium substituted axitinib.

2. The compound of claim 1, wherein: the seleno-axitinib is an antitumor drug.

3. A compound according to claim 1 or 2, wherein: the structure of the seleno-axitinib is as follows:

4. a compound according to claim 1 or 3, characterized in that: the synthetic steps of the seleno-axitinib are as follows:

5. the antitumor agent as claimed in claim 1 or 3, wherein: it can be administered by various routes, such as oral, arterial, subcutaneous, intramuscular, intraluminal, peritumoral, etc.

6. The antitumor agent as claimed in claim 1 or 3, wherein the administration route and the administration dose are varied depending on the administration subject, the administration route or the formulation form of the agent, provided that the composition can achieve an effective blood concentration in the mammal.

7. The use of the antitumor agent according to claim 1 or 3 as a therapeutic agent for kidney cancer or as a clinically adjunctive therapeutic agent for malignant tumor.