Active oxygen response type local anesthetic based on quaternary amine lidocaine derivative as well as preparation and application of active oxygen response type local anesthetic

Abstract

The invention belongs to the field of medicines, and particularly relates to an active oxygen response type local anesthetic based on a quaternary amine lidocaine derivative as well as preparation and application of the active oxygen response type local anesthetic. The medicine disclosed by the invention can selectively generate an anesthetic effect in a living body for a long time.

Description

Technical Field

[0001] The present invention belongs to the field of medicines, and in particular relates to an active oxygen responsive local anesthetic drug based on a quaternary lidocaine derivative, and a preparation method and application thereof. Background Art

[0002] Compared with systemic analgesics, local anesthetics can show obvious advantages in relieving pain and reducing adverse reactions. However, its short-term effect often limits its clinical application, so it is necessary to develop local anesthetics with long-term effect, selective action and low toxicity (Wang Q, Zhang Y, Liu J, et al. Quaternary lidocaine derivatives: past, present, and future [J]. Drug Design, Development and Therapy, 2021: 195-207.). Quaternary ammonium lidocaine derivatives (QLD) have been shown to have these characteristics in electrophysiological studies and are therefore considered to be a very promising local anesthetic. QLD is expected to become a hot topic in the research of local anesthetic drugs in the future. QLD has attracted the attention of researchers since the 1970s. It has a similar chemical structure to commonly used lidocaine, but its molecule contains a quaternary ammonium salt group, which gives QLD a longer duration of anesthesia and a more selective local anesthetic effect. QLD has a wide range of applications, such as clinical medicine, surgery, and pain management. Because its mode of action is local anesthesia, it can effectively relieve pain and reduce the adverse reactions caused by general anesthetics. At the same time, the long-term effect of QLD can shorten the time for patients to recover after surgery and reduce unnecessary medical expenses. Nowadays, the main types of QLD include QX-222, QX-572, QX-314 and QX-OH, etc. Among them, QX-314, as a local anesthetic currently being widely studied, has advantages such as longer duration and selective sensory blockade compared with traditional local anesthetics (Lim TK, Macleod BA, Ries CR, et al. The quaternary lidocaine derivative, QX-314, produces long-lasting local anesthesia in animal models in vivo [J]. The Journal of the American Society of Anesthesiologists, 2007, 107: 305-311.). However, QX-314 still has some limitations. Because it has a very strong dose dependence, serious toxic and side effects will occur when the dose is too high, so strict dose control is required. Moreover, the reduced lipophilicity of QX-314 makes it difficult for it to enter neural cells to exert its effects. Therefore, it is imperative to overcome these limitations.

[0003] Quaternary ammonium lidocaine derivatives such as QX-314 and QX-222 have the characteristics of long-term nerve conduction blockade. Among them, QX-314 has attracted widespread attention because, in addition to producing long-term peripheral nerve block, it also has sensory selective analgesia when used in combination with capsaicin, chemical permeation enhancers, etc. However, the researchers used lidocaine (70mM) as a positive control group and saline as a negative control group to investigate the adverse reactions of QX-314, and found that all mice injected with saline and lidocaine behaved normally. In contrast, in the QX-314 injection group, at a dose of 5mM, 4 of 6 mice (67%) produced motor blockade and 5 (83%) produced sensory blockade, while at a dose of 10mM, 5 of 6 mice (83%) produced sensory blockade and 6 of 6 mice (100%) showed obvious irritation reactions. It is worth noting that one of the six mice injected with a 5mM dose (17%) died, and two of the six mice injected with a 10mM dose (33%) died. The results show that although QX-314 can produce lasting nerve blockade, it is unlikely to be a drug suitable for human spinal anesthesia because obvious toxic reactions have already occurred in mouse experiments (Schwarz SK, Cheung H MC, Ries CR, et al. Lumbar intrathecal administration of the quaternary lidocaine derivative, QX-314, produces irritation and death in mice [J]. The Journal of the American Society of Anesthesiologists, 2010, 113: 438-444.). In addition, the study found that two of the four mice injected with 150mM QX-314 had nerve blockade, which did not disappear during the observation period of up to one week. This shows that high concentrations of QX-314 administration can lead to irreversible nerve blockade and pose a risk of neurotoxicity (Sagie I, Kohane DS. Prolonged sensory-selective nerve blockade [J]. Proceedings of the National Academy of Sciences, 2010, 107: 3740-3745.). Systemic administration is even more likely to cause severe central nervous system and cardiac toxicity. The researchers injected different doses of lidocaine or QX-314 (dose range 7.5-30 mg / kg) intravenously and observed signs of central nervous system toxicity in mice (convulsions, disorders, loss of righting reflex and / or death). At the same time, the electrocardiogram of the mice was also measured to evaluate the cardiotoxicity of the drug.According to the statistical analysis of experimental data, the ED50 of lidocaine for central nervous system toxicity is 19.5 mg / kg, and the ED50 of QX-314 is 10.7 mg / kg, which is lower than lidocaine. Similarly, the ED50 of lidocaine for cardiac toxicity is also higher than the ED50 of QX-314. These results show that the relative potency of QX-314 in central nervous system and heart toxicity is significantly higher than that of lidocaine at the same dose, and QX-314 is not safer than lidocaine in terms of systemic toxicity. Although QX-314 may be a potentially clinically useful drug, its systemic toxicity does not make it an advantage in use compared to traditional local anesthetics. Summary of the invention

[0004] In view of the shortcomings of the existing problems, the purpose of the present invention is to provide an active oxygen responsive local anesthetic based on a quaternary lidocaine derivative and its preparation and use.

[0005] The technical solution adopted by the present invention to solve the technical problem is:

[0006] In the first aspect, the present invention protects a compound S-QXOH, which has the following structure:

[0007]

[0008] When the above compounds come into contact with active oxygen, the sulfur atom in the molecule is oxidized by the active oxygen, and the oxygen atom of the active oxygen further attacks the ester bond in the molecule, thereby breaking the ester bond and releasing the local anesthetic QX-OH molecule to produce an anesthetic effect.

[0009]

[0010] In the second aspect, the present invention protects the preparation method of the aforementioned compound, and its synthetic route is as follows:

[0011]

[0012] The specific preparation method is as follows:

[0013] Step 1, synthesis of compound QX-OH: add lidocaine and 2-bromoethanol to a reaction bottle, heat to 100±5°C for reaction, add ethyl acetate to the reaction solution after the reaction is completed, stir for 0.5±0.5h, filter and wash to obtain compound QX-OH;

[0014] Step 2, synthesis of compound Ace-QXOH: under nitrogen protection, compound QX-OH was dissolved in N,N-dimethylformamide, triethylamine was added to the reaction solution, and the mixture was stirred under an ice bath. After the reaction solution was cooled, acryloyl chloride was slowly added dropwise to the reaction solution, and the mixture was reacted at room temperature. After the reaction was completed, ice water was added to the reaction solution to quench the reaction, the aqueous phase was extracted, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified to obtain compound Ace-QXOH;

[0015] Step 3, synthesis of compound S-QXOH: dissolve compound Ace-QXOH in acetonitrile, add acetylcysteine ​​and triethylamine to the reaction solution in sequence, stir at room temperature for 5±5 min, evaporate the solvent, and purify the crude product to obtain compound S-QXOH.

[0016] In a third aspect, the present invention protects the use of the above-mentioned compound in the preparation of local anesthetic drugs.

[0017] In a specific embodiment, the anesthetic drug is a local anesthetic drug.

[0018] Preferably, the local anesthetic is a long-acting local anesthetic or a selective local anesthetic.

[0019] In a fourth aspect, the present invention protects a pharmaceutical composition comprising the compound described above.

[0020] In a specific embodiment, the pharmaceutical composition further contains a pharmaceutically acceptable carrier or excipient.

[0021] Preferably, the pharmaceutically acceptable carrier or excipient is selected from sodium bicarbonate and physiological saline or water for injection.

[0022] In a specific embodiment, the pharmaceutical composition includes an oral preparation, an injection preparation, a transdermal preparation or a mucosal preparation.

[0023] The present invention establishes a rat sciatic nerve block model, takes 0.6 mL of compound S-QXOH (30 mM), injects the rat's sciatic nerve, and the rat's pain sensation is normal 10 minutes after the injection. Take 0.3 mL of 3% H2O2, inject the rat's sciatic nerve. After 10 minutes of injection of 3% H2O2, the rat's pain sensation is completely blocked, and the pain sensation is still completely blocked after 60 minutes. The experimental results show that the compound of the present invention can cause long-term sensory block in rats.

[0024] Beneficial Effects

[0025] The present invention provides an active oxygen responsive local anesthetic based on a quaternary lidocaine derivative, which can produce a long-lasting and low-toxic anesthetic effect in vivo and is suitable for popularization and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The results are shown for the sciatic nerve injection of 0.6 mL of compound S-QXOH (30 mM); the 10 min time point is the time of hydrogen peroxide injection.

[0027] Figure 2 The survival rate of C2C12 mouse myoblasts incubated with different concentrations of lidocaine, compound QX314 and S-QXOH.

[0028] Figure 3 The survival rate of PC12 rat adrenal pheochromocytoma cells incubated with different concentrations of lidocaine, compound QX314 and S-QXOH.

[0029] Figure 4 The staining results of muscle samples after injection of compound QX314 and S-QXOH.

[0030] Figure 5 The staining results of sciatic nerve after injection of compound S-QXOH. DETAILED DESCRIPTION

[0031] The present invention is further described in detail below with reference to the examples. The reagents or instruments used without indicating the manufacturer are regarded as conventional products that can be purchased on the market.

[0032] Example 1: Synthesis of compound S-QXOH

[0033]

[0034] F1: Synthesis of compound QX-OH. Lidocaine (5 g, 21.3 mmol) and 2-bromoethanol (1.5 mL, 21.3 mmol) were added to a reaction bottle and heated to 100°C for reaction. After the reaction was completed, 500 mL of ethyl acetate was added to the reaction solution and stirred for 0.5 h. 7.3 g of QX-OH was obtained by filtration and washing as a white solid with a yield of 96%.

[0035] 1 H NMR (300MHz, Methanol-d4) δ7.14–7.07(m,3H),4.52(s,2H),4.07–3.98(m,2H),3.85–3.65(m,6H),2.24(s,6H),1.39(t,J=7.2Hz,6H). 13C NMR (75MHz, Methanol-d4) δ164.02,136.74,134.13,129.22,128.83,61.53,58.44,56.89,56.69,18.67,8.31.

[0036] F2: Synthesis of compound Ace-QXOH. Under nitrogen protection, QX-OH (5 g, 14 mmol) was dissolved in 30 mL of N,N-dimethylformamide. Triethylamine (2.9 mL, 21 mmol) was added to the reaction solution and stirred under an ice bath. After the reaction solution was cooled, acryloyl chloride (1.36 mL, 16.8 mmol) was slowly added dropwise to the reaction solution, and the ice bath was removed to allow the reaction solution to react at room temperature. After the reaction was completed, ice water was added to the reaction solution to quench the reaction, and the aqueous phase was extracted with dichloromethane. The organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to obtain 3.5 g of compound Ace-QXOH with a yield of 75.3%.

[0037] 1 H NMR (300MHz, DMSO-d6) δ10.20(s,1H),7.10(d,J=1.8Hz,3H),6.38(dd,J=17.2,1.7Hz,1H),6.23(dd,J=17.2,10.2Hz,1H),6.03(dd,J= 10.2,1.7Hz,1H),4.60(t,J=4.9Hz,2H),4.48(s,2H),3.97(t,J=5.0Hz,2H),3.65(q,J=7.1Hz,4H),2.16(s,6H),1.33(t,J=7.1Hz,6H).

[0038] F3: Synthesis of compound S-QXOH. Compound Ace-QXOH (0.5 g, 1.5 mmol) was dissolved in 5 mL of acetonitrile, and acetylcysteine ​​(0.24 g, 1.8 mmol) and triethylamine (0.31 mL, 2.2 mmol) were added to the reaction solution in sequence, and stirred at room temperature for 5 min. The solvent was evaporated, and the crude product was purified by column chromatography to obtain 0.7 g of compound S-QXOH with a yield of 94%.

[0039] 1H NMR(300MHz, Methanol-d4)δ7.08(d,J=3.8Hz,3H),4.59–4.54(m,1H),4.50(s,2H),4.38(dd,J=6.7,4.5Hz,1H),3.98(t,J=5.1Hz,2H),3 .76(q,J=7.2Hz,4H),3.18–2.99(m,2H),2.86–2.79(m,2H),2.69(dd,J=7.2,5.7Hz,2H),2.22(s,6H),1.94(s,3H),1.41(t,J=7.2Hz,6H).

[0040] Example 2: In vivo activity evaluation experiment

[0041] A rat sciatic nerve block model was established, and a hot plate test was used to determine whether the rats had sensory block. The rats in the control group were injected with 0.6 mL of 3% hydrogen peroxide solution. After 10 minutes and 30 minutes of injection, neither sensory nor motor block occurred. 0.6 mL of compound S-QXOH (30 mM) was injected into the sciatic nerve of the rats. After 10 minutes of injection, no sensory block occurred in the rats. 0.3 mL of 3% hydrogen peroxide solution was injected 10 minutes after the injection of 0.6 mL of compound S-QXOH (30 mM). After 10 minutes, the sensory nerves of the rats were completely blocked, and the sensory block still existed after 60 minutes. The experimental results show that the compounds of the present invention can cause long-term sensory block in rats.

[0042] Example 3: Cytotoxicity Experiment

[0043] The toxicity of S-QXOH to muscle cells and neurons was evaluated using C2C12 mouse myoblasts (American Type Culture Collection (ATCC), Manassas, VA, USA) and PC12 rat adrenal pheochromocytoma cells (ATCC, Manassas, VA, USA), respectively.

[0044] C2C12 cells were seeded at a density of 1,000 cells per well in DMEM containing 20% ​​FBS and 1% penicillin-streptomycin and then incubated in DMEM containing 2% horse serum and 1% penicillin-streptomycin for 10-14 days to differentiate into myotubes. PC12 cells were seeded at a density of 4,000 cells per well in DMEM containing 2.5% FBS, 12.5% ​​horse serum and 1% penicillin-streptomycin and then incubated in DMEM containing 1% horse serum, 50 ng / mL nerve growth factor and 1% penicillin-streptomycin for 7 days. The cytotoxicity of S-QXOH was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) colorimetric method. C2C12 cells and PC12 cells were treated with different concentrations of compound S-QXOH and compound QX-314, respectively, and lidocaine (a local anesthetic commonly used in clinical practice) was used as a positive control drug. After incubation for 24 hours, 40 μL of MTS solution was added to each well, and the cells were incubated for another 4 hours. Synergy TM The absorbance (490 nm) was measured on an Mx microplate reader (BioTek Inc., VT, USA). The viability was expressed as a percentage of the result of untreated cells.

[0045] The experimental results show that ( Figure 2-Figure 3 ), the cell survival rate after incubation with compound S-QXOH was significantly higher than that after incubation with compound QX-314 and positive control lidocaine. This indicates that the cytotoxicity of compound S-QXOH is significantly lower than that of compound QX-314 and lidocaine.

[0046] Example 4: Muscle and nerve tissue toxicity test

[0047] Four days and fourteen days after rats were injected with 0.6 mL of compound S-QXOH (30 mM) and 0.6 mL of compound QX-314 (30 mM), respectively, the animals were euthanized by inhalation of carbon dioxide to evaluate the inflammation and toxicity of compound S-QXOH and compound QX-314. The sciatic nerve and adjacent connective tissue closest to the nerve block injection site were obtained using sterilized surgical instruments. The muscle samples were fixed in 10% neutral buffered formalin solution, and the muscle tissue was embedded in paraffin. After sectioning, the muscle was stained with hematoxylin and eosin using standard techniques. Each section was examined in a blind manner using an optical microscope. The rat sciatic nerve was fixed in Karnovsky's KII solution, treated accordingly, embedded in resin, and stained with toluidine blue. In order to ensure the objectivity of the experimental results, the experimenter evaluated in a blind manner using an optical microscope. The experimental results show that ( Figure 4-Figure 5), the muscle toxicity of compound S-QXOH is less than that of compound QX-314, and compound S-QXOH has no neurotoxicity.

[0048] The protection content of the present invention is not limited to the above embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be thought of by those skilled in the art are included in the present invention and are protected by the attached claims.

Claims

1. A compound having the following structure:

2. The method for preparing the compound according to claim 1, characterized in that: Its synthetic route is as follows: The preparation method is as follows: Step 1, synthesis of compound QX-OH: add lidocaine and 2-bromoethanol to a reaction bottle, heat to 100±5°C for reaction, add ethyl acetate to the reaction solution after the reaction is completed, stir for 0.5±0.5h, filter and wash to obtain compound QX-OH; Step 2, synthesis of compound Ace-QXOH: under nitrogen protection, compound QX-OH was dissolved in N,N-dimethylformamide, triethylamine was added to the reaction solution, and the mixture was stirred under an ice bath. After the reaction solution was cooled, acryloyl chloride was slowly added dropwise to the reaction solution, and the mixture was reacted at room temperature. After the reaction was completed, ice water was added to the reaction solution to quench the reaction, the aqueous phase was extracted, the organic phase was collected, dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified to obtain compound Ace-QXOH; Step 3, synthesis of compound S-QXOH: dissolve compound Ace-QXOH in acetonitrile, add acetylcysteine ​​and triethylamine to the reaction solution in sequence, stir at room temperature for 5±5 min, evaporate the solvent, and purify the crude product to obtain compound S-QXOH.

3. Use of the compound according to claim 1 in the preparation of anesthetic drugs.

4. The use according to claim 3, characterized in that: The anesthetic drug is a local anesthetic drug.

5. The use according to claim 4, characterized in that: The local anesthetic is a long-acting local anesthetic or a selective local anesthetic.

6. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the compound according to claim 1.

7. The pharmaceutical composition according to claim 6, characterized in that The pharmaceutical composition also contains pharmaceutically acceptable carriers or excipients.

8. The pharmaceutical composition according to claim 6, characterized in that The pharmaceutically acceptable carrier or excipient is selected from sodium bicarbonate and physiological saline or water for injection.

9. The pharmaceutical composition according to any one of claims 6 to 8, characterized in that The pharmaceutical composition includes oral preparations, injection preparations, transdermal preparations or mucosal preparations.

Images