WO2017156648A1

WO2017156648A1 - Compound having anti-inflammatory effect

Info

Publication number: WO2017156648A1
Application number: PCT/CN2016/000142
Authority: WO
Inventors: 陈秀兰
Original assignee: 广州诺威生物技术有限公司
Priority date: 2016-03-18
Filing date: 2016-03-18
Publication date: 2017-09-21

Abstract

A compound having anti-inflammatory effect. The compound has a structure as shown in formula (I), and has a stronger anti-inflammatory and analgesic effect comparing to ibuprofen.

Description

An anti-inflammatory compound

Technical field

The present invention relates to a novel compound having an anti-inflammatory action.

Background technique

Antipyretic analgesic and anti-inflammatory drugs are a class of drugs that have antipyretic and analgesic effects, and most have anti-inflammatory and anti-rheumatic effects. Because of its different chemical structure and anti-inflammatory mechanism than glucocorticoid steroidal anti-inflammatory drugs (SAIDS), different types of NSAIDs, also known as non-steroidal anti-inflammatory drugs (NSAIDS), have the same mechanism of action. They inhibit the final production of prostacyclin (PGI1), prostaglandins (PGE1, PGE2) and thromboxane A2 (TXA2) by inhibiting the activity of cyclooxygenase. Prostaglandins have many functions: increased vascular permeability; various tissue arterial dilatation; regulation of renal blood flow, increased renal filtration rate; promotion of sodium excretion, lowering blood pressure; inhibition of gastric acid secretion; contraction of uterine muscle fibers, dissolution of the corpus luteum; Relaxation of tracheal smooth muscle; vasoconstriction of nasal mucosa; inhibition of platelet aggregation; promotion of bone resorption; inhibition of glycerolipid decomposition. In addition to inhibiting the synthesis of prostaglandins, NSAID also inhibits the release of bradykinin during inflammation, alters lymphocyte responses, and reduces the migration and phagocytosis of granulocytes and monocytes. It is also because NSAID inhibits the synthesis of prostaglandins, in addition to the analgesic and anti-inflammatory effects, there are also corresponding side effects. Mainly in the gastrointestinal tract and kidneys. To evaluate the efficacy of NSAIDs, most NSAIDs were compared to ibuprofen or aspirin.

Summary of the invention

The present invention relates to a novel compound SZY1406 (formula I), as shown in the following figure:

Non-steroidal anti-inflammatory drugs have anti-inflammatory, analgesic and antipyretic effects; the experimental injection of carrageenan replicates the inflammatory model, and the multi-dose group is established to observe the effect of oral administration of SZY1406 and ibuprofen on paw swelling in rats. The strength of SZY1406 and ibuprofen. Normal toe volume measured before dosing, grouped by volume, 10 per group Only, once a dose, immediately after the administration of the toe injection of 1% carrageenan 0.1ml / only, measured the toe volume of rats at 1h, 2h, 3h, 4h, 5h after inflammation, statistical toe before and after inflammation Volume and rate of swelling of the toes after inflammation, and comparative analysis of the intensity of ibuprofen and SZY1406. Compared with before administration, ibuprofen 2mg/kg showed a significant increase in the toe volume at each time point after administration, suggesting that ibuprofen 2mg/kg has no anti-inflammatory effect on carrageenan-induced inflammation, and cloth There was no increase in the volume of rat toe at 8, 12, and 16 mg/kg of loffin at each time point after administration; SZY1406 2 mg/kg increased the volume of rat toe after administration, 2 h, 3 h after administration. The increase was more obvious. SZY1406 4, 8 mg/kg showed no increase in rat toe volume at each time point after administration. Compared with the model group, the swelling rate of the toes in the rats with ibuprofen at 8, 12, and 16 mg/kg was significantly decreased. The swelling rate of the toes in SZY14062, 4, 8, 12, and 16 mg/kg rats decreased to varying degrees. The toe swelling rate of the rats in the dose group was significantly reduced 5 h after administration. According to the changes of rat toe volume and swelling rate before and after administration, it can be seen that under the experimental conditions, ibuprofen and SZY1406 have obvious anti-inflammatory effects, and ibuprofen can obtain stable anti-inflammatory effect at 8mg/kg. SZY1406 2mg/kg can obtain strong anti-inflammatory effect 5h after administration.

SZY1406 is an anti-inflammatory and analgesic non-steroidal anti-inflammatory drug. The carrageenan inflammation model used in the experiment is to induce the inflammation of the carrageenan in the animal to cause inflammation or a certain stage of cellular tissue reaction. It can be seen from the experiment that the injection of carrageenan can cause obvious inflammatory reaction in the rat toe. After the intervention of the test substance, the ibuprofen suspension can obtain stable anti-inflammatory effect at 8 mg/kg, SZY1406 The swelling rate of the toe in the dose group was reduced at different levels at 1h, 2h, 3h and 4h after administration. 2mg/kg could exert a good anti-inflammatory effect, and the toe swelling of the rat was 5h after administration. The rate is significantly reduced, showing a strong anti-inflammatory effect, suggesting that the anti-inflammatory effect of SZY1406 takes a long time, and the anti-inflammatory effect is stronger than that of ibuprofen.

Specific embodiment

Example 1: Preparation of SZY1406 The synthetic route is as follows:

Specific synthesis method:

Compound 1 (11.2 g, 0.05 mol), Compound 2 (11.6 g, 0.05 mol), HOBT (7.5 g, 0.055 mol) and N-methylmorpholine (16.5 ml, 0.15 mol) were dissolved in a nitrogen atmosphere. In chloroform (150 ml), EDCI (10.6 g, 0.055 mol) in DMF (50 ml) was added dropwise under ice bath. After the completion of the dropwise addition, the reaction was carried out at 0 ° C for 30 min, and the reaction was carried out at room temperature for 6 h. The reaction was completed by TLC (DCM / MeOH = 10/1). The organic phase was washed twice with dilute hydrochloric acid, saturated brine and water. The product was dried, and the solvent was evaporated to dryness (DCM/MeOH = 200/1 to 50/1) to afford product (yield: 90%).

Compound 3 (18 g, 0.045 mol), compound 4 (9.54 g, 0.045 mol) and DMAP (0.6 g, 4.95 mmol) were dissolved in dichloromethane (100 ml) under nitrogen atmosphere. EDCI (9.5 g, 0.0495 mol). After the reaction was carried out at 0 ° C for 30 min, the reaction was carried out at room temperature for 4 h, and the reaction was completed by TLC (DCM / MeOH = 30/1). The organic phase was washed twice with dilute hydrochloric acid, saturated sodium bicarbonate and water. After drying, the solvent was evaporated to dryness (DCM / MeOH = 200/1 to 100/1) to afford product 23.1 g (yield 88.5%).

Compound 5 (23.1 g, 0.04 mol) and 10% Pd/C (2 g) were dissolved in acetic acid (50 ml) under a hydrogen atmosphere, and reacted at room temperature for 24 h. The reaction was completed by TLC (MeOH) and filtered through celite. Pd/C, the solvent was distilled off (DCM/MeOH = 5/1 to 1/10) to give a product (13.6 g, purity ≥97%, yield 95.5%). The NMR spectrum is shown in Figure 1.

Example 2: SZY1406 pharmacodynamic experiment

1 SZY1406 on the pain caused by hot plate method in mice

Female Kunming mice weighing 20-22 g were used. First, the pain sensitivity screening was performed. The mice were placed on a hot plate at 55.0 °C ± 0.5 °C. The time experienced by the hind paws of the mice was used as the threshold of pain response, and the jumpers were removed and less than 5 s or more than 30 s. None of the responders were screened for 60 pain-sensitive mice, which were randomly divided into five groups according to body weight, with 12 rats in each group. Experiments were performed using SZY1406. SZY1406 low, medium and high dose groups (SZY1406 doses of 4, 12, 36mg / kg body weight) and positive control group (ibuprofen, dose of 240mg / kg body weight), using 0.5% sodium carboxymethyl cellulose The solution is prepared as a suspension. After the pre-dose pain threshold was measured, the mice in each test group were intragastrically administered with the corresponding drugs, and the blank control group was given an equal volume of physiological saline. The hairs of the mice were removed from the ankle joints of the hind paws with a depilatory agent and dried. The pre-dose pain response threshold was determined in the same manner for each mouse, and then the pain response threshold after each rat administration was measured 1 h after each test group, and the mouse was immediately removed from the hot plate after the hind paw. The threshold of the pain response was measured again 2 h and 3 h after the administration, and the threshold of the pain response between the groups was subjected to t test to evaluate the analgesic effect. The results of the test are shown in Table 1 and Figure 1. The results show that the thresholds of the pain response of the hot plate method before the administration of the mice in each test group are similar. After the administration, the mice in each dose group of SZY1406 are licked at 1h, 2h, 3h. The time was significantly longer than that of the blank control group, which increased the threshold of pain response, indicating that SZY1406 can significantly prolong the pain threshold of hot plate in mice, has significant analgesic effect, and is significantly better than ibuprofen. In Table 1, the low, medium, and high dose groups represent the SZY1406 low, medium, and high dose groups, respectively.

Table 1 Effect of SZY1406 on the pain threshold of mice in hot plate assay

Note: Compared with the blank control group: *P<0.05; **P<0.01.

2. The effect of SZY1406 on carrageenan-induced paw swelling in rats

Self-made rat toe volume measuring instrument: Take 2 ml, 20 ml syringes, 1 three-way piston, 1 pipette, 1 infusion tube, and install according to Figure 2. At the top of the 20ml syringe housing, use a black line to circumscribe the horizontal mark as a horizontal line. Rotate the tee to make the pipette communicate with the 2ml syringe. Push the syringe to make the liquid level of the pipette at 0 mark. Rotate the tee to make the 2ml syringe and 20ml. The syringe is connected, the liquid in the 2ml syringe is pushed out, and the liquid level in the syringe of 20ml is adjusted to the horizontal line with a straw. Rat toe volume measurement can be performed.

When measuring, draw a horizontal line on the upper part of the upper leg of the rat's toe ankle with a marker, grab the rat, gently press the hip joint, keep the hind limb of the rat stretched, and extend the rat toe into the 20ml syringe. In the outer casing, adjust the height of the rat so that the toe line coincides with the horizontal line on the 20 ml syringe. At the same time, the liquid level in the 20 ml syringe is controlled by a 2 ml syringe to coincide with the horizontal line and the rat toe line. Remove the rat, put it back into the cage, rotate the tee, connect the 2 ml syringe to the pipette, push the liquid from the 2 ml syringe into the pipette, and read the volume, which is the rat toe volume. After reading the volume, adjust the liquid level of the pipette to 0 scale, rotate the tee, connect the 2ml syringe to the 20ml syringe casing, push out the 2ml inner liquid, and adjust the liquid level of the 20ml syringe casing with the straw to coincide with the horizontal line. A toe volume measurement of a rat.

50 rats weighing 220-250g were used, and after 12 hours of fasting, the right hind foot toe volume of all animals was measured as the base volume using a self-made rat toe volume tester. After the measurement, the animals were placed according to the toe volume. They were randomly divided into 5 groups, SZY1406 low, medium and high dose groups (SZY1406 doses were 3.2, 9.6, 28.8 mg/kg body weight, respectively), and another vehicle control group and positive control group (ibuprofen, dose 192 mg/kg). Body weight), 10 rats in each group, the test substance was prepared by using 0.5% sodium carboxymethyl cellulose as a suspension. Immediately after grouping, 1% carrageenan 0.1ml/only was injected subcutaneously into the right hind foot and ankle. Immediately after the injection of carrageenan, the test substance was intragastrically administered once, and the right hind paw toe volume of the rats at 1 h, 2 h, and 3 h after administration was measured, and the swelling rate of the toe of the rat was calculated, and the swelling rate (%) was calculated. = toe volume after inflammation * 100% / volume before inflammation.

The results of the test showed that the swelling rate of the toes in the rats after inflammation was calculated by the formula: swelling rate (%) = volume of the toes after inflammation * 100% / volume of the toes before inflammation, from the results, the model control group was inflamed The swelling rate of the toe of the rat was significantly increased. Compared with the model group, the swelling rate of the toe in the low dose group of SZY1406 was decreased at 1h, 2h, 3h after administration, and the toe of SZY1406 medium and high dose group was rat. The swelling rate was significantly decreased at 1 h, 2 h, and 3 h after administration (P < 0.01). Compared with before administration, the model control group showed a significant increase in the toe volume of the rat after injection of carrageenan (P<0.01). The low dose of SZY1406 did not increase as much as the model control group after administration. Compared with before administration, the increase was significant (P<0.01). The volume of the toe in the middle and high dose groups of SZY1406 did not increase significantly after administration, suggesting that SZY1406 medium and high doses were induced by carrageenan. Rat paw swelling has obvious inhibitory effects. The results are shown in Table 2 and Table 3.

Table 2 Effect of swelling rate on the toe of rats after inflammation (%)

Note: Compared with the model control group: **P<0.01.

Table 3 Effect of before and after toe volume on rats before and after inflammation (ml)

Note: Compared with before administration: **P<0.01.

According to the change of rat toe volume and swelling rate before and after administration, it can be seen that under the experimental conditions, both ibuprofen and SZY1406 have obvious anti-inflammatory effects, and the anti-inflammatory effect of SZY1406 is much greater than that of ibuprofen.

Claims

a compound of the structure shown in formula (I):
A salt of the compound of claim 1.
The sodium salt of the compound of claim 1.
Use of a compound according to claims 1-3 for the preparation of an anti-inflammatory analgesic.