Compound with local anesthesia function as well as preparation method and application thereof

Abstract

The invention discloses a compound with a local anesthesia function as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The preparation method of the compound comprises the following steps: mixing and heating lidocaine and 2-bromoethanol to obtain a compound 1; the preparation method comprises the following steps: mixing dicarboxylic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and a solvent to obtain a mixed solution; mixing and dissolving a compound 1, alkali and a solvent, and then adding into the mixed solution; and then carrying out spin-drying, separation and purification to obtain the compound with the local anesthesia function. The medicine can be used as a local anesthetic or analgesic for long-acting nerve block or selective nerve block. The medicine can block nerve conduction, has biocompatibility and biodegradability, is expected to overcome the problems of short time efficiency of single-needle injection and the like, and improves the efficacy of the medicine.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to compounds with local anesthetic function, their preparation methods, and applications. Background Technology

[0002] Commonly used local anesthetics are as follows: The aforementioned local anesthetics are the main medications used in local anesthesia. As one of the means of perioperative analgesia, local anesthesia exerts its analgesic effect by injecting local anesthetics around the epineurium, thereby blocking the signal transmission of nerve pathways. Compared with general anesthesia, which has a higher incidence of side effects (respiratory depression, addiction, etc.), local anesthesia has gradually taken a leading position in patient pain management due to its fewer side effects, lower cost, and excellent analgesic effect. However, the application of local anesthetics still faces many problems, such as the short duration of single-needle injection (<16 hours) and catheter-related complications (infection, catheter displacement) involved in continuous infusion. The short-acting nerve block time of local anesthetics and adverse reactions caused by drug overdose have always been serious problems faced by local anesthesia in clinical practice. Therefore, the development of new local anesthetics that are inexpensive, highly reproducible, easy to produce, and easy to clinically translate is of great significance and is the fastest and most effective solution to the current serious problems. Summary of the Invention

[0003] In view of the shortcomings of the prior art, the purpose of this invention is to provide a compound with local anesthetic function, its preparation method and application, thereby solving the problems in the prior art.

[0004] The objective of this invention can be achieved through the following technical solutions: The compound possessing local anesthetic function has the following structural formula: Where n is 3, 5 or 7.

[0005] A method for preparing a compound with local anesthetic function includes the following steps: S1, lidocaine and 2-bromoethanol were mixed and heated to obtain compound 1; S2, a mixed solution is obtained by mixing a dicarboxylic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride and a solvent; compound 1, a base and a solvent are mixed and dissolved, and then added to the mixed solution; after rotary evaporation, separation and purification, a compound with local anesthetic function is obtained.

[0006] Furthermore, the synthetic route for compounds possessing local anesthetic function is as follows: .

[0007] Furthermore, the structural formula of compound 1 is as follows: .

[0008] Furthermore, the solvent is tetrahydrofuran or dichloromethane.

[0009] Furthermore, the base is any one of DMAP, DIPEA, pyridine, triethylamine, and Na2CO3.

[0010] Furthermore, the dicarboxylic acid is any one of glutaric acid, heptaic acid, and azelaic acid.

[0011] Furthermore, the molar ratio of the dicarboxylic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, compound 1, and the base is 1:2.4:2.4:2.4:2.4.

[0012] The above-mentioned compounds are used in the preparation of anesthetic or analgesic drugs.

[0013] A drug characterized by comprising capsaicin and the aforementioned compounds.

[0014] The beneficial effects of this invention are: 1. The compounds prepared by this invention can be used as long-acting local anesthetics or analgesics for nerve blockade; they can block nerve conduction, have biocompatibility and biodegradability, and are expected to overcome the problems of short duration of single-injection and improve drug efficacy.

[0015] 2. The compounds prepared by this invention can be used in combination with capsaicin, which can not only serve as long-acting local anesthetic agents for nerve block, but also for selective nerve block.

[0016] 3. The compounds prepared in this invention have the effect of separating sensory and motor blockade, and can be used as tool drugs for the study of local anesthesia mechanisms.

[0017] 4. The compounds prepared by this invention, especially the DT-A002 system, have low toxicity and are expected to be translated into clinical applications.

[0018] 5. The optimal base used in the preparation of the compound in this invention is DMAP. DMAP is a common solvent for activating hydroxyl groups and is widely used in reactions such as esterification, amidation, and cyclization. DMAP-catalyzed reactions have fast rates and high selectivity, and can also improve the intermolecular forces of the catalyst under reaction conditions, making the reaction more uniform. By screening the types of bases used in the reaction process, DMAP was ultimately determined to be the optimal base. Attached Figure Description

[0019] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, for those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0020] Figure 1 The hydrogen NMR spectrum of Example 1; Figure 2 The carbon NMR spectrum of Example 1; Figure 3 The mass spectrum of Example 1; Figure 4 The hydrogen NMR spectrum of Example 2; Figure 5 The carbon NMR spectrum of Example 2; Figure 6 This is the mass spectrum of Example 2; Figure 7 The hydrogen NMR spectrum of Example 3; Figure 8 The carbon NMR spectrum of Example 3; Figure 9 This is the mass spectrum of Example 3; Figure 10 The images show hematoxylin-eosin (H&E) stained sections and magnified local sections of the sciatic nerve and surrounding muscle tissue after administration of different drug groups in Example 4. Figure 11 The images show magnified sections of sciatic nerve stained with toluidine blue (TB) and local sections after administration of different drug groups in Example 4. Figure 12 The images show hematoxylin-eosin (H&E) stained sections and magnified local sections of the sciatic nerve and surrounding muscle tissue after administration of different drug groups in Example 5, as well as toluidine blue (TB) stained sections and magnified local sections of the sciatic nerve. Figure 13 Changes in body weight 14 days after administration of different concentrations of DT-A002; Figure 14 Organ coefficients (n≥5) after 14 days of administration of different concentrations of DT-A002. Figure 15 H&E stained sections of the heart, liver, spleen, lungs, kidneys and brain of mice in each group (n≥3), scale bars=50 μm. Detailed Implementation

[0021] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] The synthetic route for compounds with local anesthetic function is as follows: The process for preparing the target product is as follows: S1, lidocaine was mixed with 2-bromoethanol and heated to give compound 1 (QX-OH). S2, compound 2 (dicarboxylic acid), N-hydroxysuccinimide (NHS), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) and solvent are mixed to obtain a mixed solution; compound 1, base and solvent are mixed and dissolved, and then added to the above mixed solution; then the target product 1 is obtained by rotary evaporation, separation and purification.

[0023] The solvent is tetrahydrofuran or dichloromethane; the base used is any one of DMAP, DIPEA, pyridine, triethylamine, or Na2CO3; compound 2 (dicarboxylic acid) is any one of glutaric acid, pimelic acid, or azelaic acid.

[0024] The molar ratio of compound 2, NHS, EDCI, compound 1 and base is 1:2.4:2.4:2.4:2.4.

[0025] The technical solution of the present invention will be described below through the following embodiments; the following embodiments use lidocaine, which is commonly used in clinical practice, as the initial raw material for the reaction, and the structure of compound 1 is shown below: .

[0026] Furthermore, in the embodiments, "equivalent" refers to molar parts.

[0027] In this embodiment, the sources of the relevant raw materials are as follows: Lidocaine: Manufacturer: Shanghai McLean Biochemical Technology Co., Ltd., Brand: D822738-100g; NHS: Manufacturer is Adamas Reagents Ltd., brand number is 69489B, 25g; EDCI: Manufacturer is Adamas Reagents Ltd., brand name is 42385B, 25g; DMAP: Manufacturer is Jiangsu Aikon Biomedical R&D Co., Ltd., brand name is AK0034BU, 100g; Capsaicin: Manufacturer: Hebei Bailingwei Ultrafine Materials Co., Ltd., Brand: 196918, 1g; Pyridine: Manufacturer: Adamas Reagent Co., Ltd., Brand: 14230I, 100mg; Triethylamine: Manufacturer: Adamas Reagent Co., Ltd., Brand No. 17471I, 100mL; DIPEA: Manufacturer is Shanghai Maclean Biochemical Technology Co., Ltd., brand name N807281-500mL; Na2CO3: Manufacturer: Sinopharm Chemical Reagent Co., Ltd., Brand: 20210105, Analytical Grade AR, 500g.

[0028] Example 1 ① Reaction Procedure: Weigh glutaric acid (66.0 mg, 0.50 mM, 1 equivalent), N-hydroxysuccinimide (NHS) (138.0 mg, 1.20 mM, 2.4 equivalent), and 1-ethyl-3-(3-Dimethylaminopropyl)carbodiimide hydrochloride (EDCI) (230.0 mg, 1.20 mM, 2.4 equivalent), place them in a 50 mL round-bottom flask that has been treated to remove water and oxygen, add 5 mL of methylenechloromethane (DCM), and stir at room temperature for 4 h; then weigh compound 1 (QX-OH) (430.8 mg, 1.20 mM, 2.4 equivalent) and 4-dimethylaminopyridine (DMAP) (146.4 mg, 1.20 mM, 1.20 mM, 2.4 equivalent). After dissolving 5 mL of DCM (mM, 2.4 equivalents) in the solution, add it to a round-bottom flask and stir at room temperature for 6 h.

[0029] ② Post-processing: Add an appropriate amount of silica gel (about 500.0 mg) to the flask, then evaporate the solvent using a rotary evaporator, and separate and purify using a rapid preparative liquid chromatograph. The eluents are methanol (MeOH) and DCM, and the elution time is 1 h.

[0030] ③ Product Analysis: Hydrogen nuclear magnetic resonance (HMR) spectroscopy and carbon nuclear magnetic resonance (CMR) spectroscopy were used to verify the structural characteristics and purity of the synthesized product. From... Figure 1 , Figure 2 and Figure 3 The image shows the NMR spectrum of DT-A001. The chemical shifts of various hydrogen atoms in the NMR spectrum of DT-A001 are as follows: 1 H NMR (400 MHz, Chloroform- dThe δ values ​​were 10.17 (s, 1H), 7.02 (s, 3H), 4.90 (s, 2H), 4.62 (s, 2H), 4.02 (s, 2H), 3.78 (s, 4H), 2.55 (s, 2H), 2.25 (s, 6H), 1.98 (s, 1H), and 1.42 (s, 6H). High-resolution mass spectra showed that the molecular ion peak of DT-A001 was at 327.22, confirming the successful synthesis of the compound. The yield was 57%.

[0031] The results of this embodiment demonstrate that the preparation method of the present invention can obtain the target product: DT-A001.

[0032] Example 2 ①Reaction process: Weigh pimelic acid (80.1 mg, 0.50 mM, 1 equivalent), NHS (138.00 mg, 1.20 mM, 2.4 equivalent), and EDCI (230.0 mg, 1.20 mM, 2.4 equivalent), place them in a 50 mL round-bottom flask that has been treated with anhydrous and oxygen-free conditions, add 5 mL of DCM and stir at room temperature for 12 h; then weigh compound 1 (QX-OH) (430.8 mg, 1.20 mM, 2.4 equivalent) and DMAP (146.4 mg, 1.20 mM, 2.4 equivalent), dissolve them in 5 mL of DCM and add them to the round-bottom flask, stir at room temperature for 12 h.

[0033] ② Post-processing: Add an appropriate amount of silica gel (about 500.0 mg) to the flask, then evaporate the solvent using a rotary evaporator, and perform separation and purification using a rapid preparative liquid chromatograph. The eluents are MeOH and DCM, and the elution time is 1 h.

[0034] ③ Product Analysis: Hydrogen nuclear magnetic resonance (HMR) spectroscopy and carbon nuclear magnetic resonance (CMR) spectroscopy were used to verify the structural characteristics and purity of the synthesized product. From... Figure 4 , Figure 5 and Figure 6 As can be seen from the NMR spectrum, the chemical shifts of various hydrogen atoms in DT-A002 are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 11.01 (s, 1H), 7.04 (s,3H), 5.06 (s, 2H), 4.71 (s, 2H), 4.03 (s, 2H), 3.77 (d, J = 7.2 Hz, 4H), 2.43(d, J= 6.7 Hz, 2H), 2.26 (s, 6H), 1.66 (s, 2H), 1.52 (d, J = 7.1 Hz, 6H), 1.44(d, J = 7.4 Hz, 1H). High-resolution mass spectra showed that the molecular ion peak of DT-A002 was at 341.24, confirming the successful synthesis of the compound. The yield was 85%.

[0035] The results of this embodiment demonstrate that the preparation method of the present invention can obtain the target product: DT-A002.

[0036] Example 3 ①Reaction process: Weigh azelaic acid (94.1 mg, 0.50 mM, 1 equivalent), NHS (138.0 mg, 1.20 mM, 2.4 equivalent), and EDCI (230.0 mg, 1.20 mM, 2.4 equivalent), place them in a 50 mL round-bottom flask that has been treated with anhydrous and oxygen-free conditions, add 5 mL of DCM and stir at room temperature for 24 h; then weigh compound 1 (QX-OH) (430.8 mg, 1.20 mM, 2.4 equivalent) and DMAP (146.4 mg, 1.20 mM, 2.4 equivalent), dissolve them in 5 mL of DCM and add them to the round-bottom flask, stir at room temperature for 24 h.

[0037] ② Post-processing: Add an appropriate amount of silica gel (about 500.0 mg) to the flask, then evaporate the solvent using a rotary evaporator, and perform separation and purification using a rapid preparative liquid chromatograph. The eluents are MeOH and DCM, and the elution time is 1 h.

[0038] ③ Product Analysis: Hydrogen nuclear magnetic resonance (HMR) spectroscopy and carbon nuclear magnetic resonance (CMR) spectroscopy were used to verify the structural characteristics and purity of the synthesized product. From... Figure 7 , Figure 8 and Figure 9 As can be seen from the NMR spectrum, the chemical shifts of various hydrogen atoms in DT-A003 are as follows: 1 H NMR (400 MHz, Chloroform- d ) δ 10.97 (s, 1H), 7.15-6.80 (m, 3H), 4.90 (s, 2H), 4.61 (d, J = 4.9 Hz, 2H), 3.98 (t, J = 4.8 Hz, 2H), 3.69(q, J = 7.1 Hz, 4H), 2.33 (q, J= 7.6 Hz, 2H), 2.24 (s, 6H), 1.60 (d, J = 6.9 Hz, 2H), 1.45 (t, J = 7.0 Hz, 6H), 1.30 (s, 2H), 1.26 (s, 1H). High-resolution mass spectra showed that the molecular ion peak of DT-A003 was 355.25, confirming the successful synthesis of the compound. Its yield was 67%.

[0039] The results of this embodiment demonstrate that the preparation method of the present invention can obtain the target product: DT-A003; that is, it can obtain a compound containing unilateral ester bonds and bilateral ester bonds.

[0040] Example 4 In this embodiment, the products prepared in Examples 1-3 and related control compounds were formulated into equimolar concentration PBS solutions and injected near the sciatic nerve. The nerve blocking effects of various small molecule drugs were evaluated using the heat shrinkage threshold test and extensor postural thrust test. A QX-OH concentration of 35 mM was selected, with 6 rats per group. The grouping of experimental subjects, administration methods, and specific procedures are shown in Table 1 below. Table 1. Drug dosage and sensorimotor blockade status in each group Experimental results showed that when the dosage of the target compound was half that of lidocaine hydrochloride and compound 1, sensory and motor blockade occurred, and the duration of both sensory and motor blockade was longer than that in the two control groups. This suggests the potential for developing long-acting local anesthetics through modifications to other dosage forms or for use as a tool in researching the mechanisms of local anesthesia. Figure 10 As shown, no inflammatory cell infiltration was observed in the target compounds DT-A001-DT-A003 or the control group, indicating that the target compounds have clinical translational potential. Based on the scoring criteria (0 = normal muscle tissue morphology; 1 = peritendinitis, but inflammatory cells did not invade deeper tissue layers (<5 cell layers); 2 = deeper inflammatory cell infiltration (>5 layers); 3 = semi-fascicular inflammatory response; 4 = whole-fascicular inflammatory response), it can be concluded that no inflammatory cell infiltration was observed in any group, indicating good biocompatibility. Figure 11 As shown, lidocaine derivatives did not exhibit any neurological damage manifestations at the same dose, including changes in cell morphology (swelling, shrinkage), cell nuclei (condensation, fragmentation, dissolution), and vacuolation.

[0041] Example 5 Based on previous experience in the development of anesthetic drugs, charged sodium channel blockers can target pain receptors via capsaicin to produce pain-specific local anesthesia. In this embodiment, DT-A002 and capsaicin were used in combination to optimize the nerve block selectivity of DT-A002. A mixed solution of 1.4 mM DT-A002 and 0.818 mM capsaicin was prepared, and the ratio of the two was appropriately adjusted. The mixed drug was injected into the sciatic nerve of rats, with 6 rats in each group, and the nerve block effect was evaluated. The sensorimotor blockade of each group is shown in Table 2 below: Table 2. Drug dosage and sensorimotor blockade status in each group As shown in Table 2, lidocaine hydrochloride combined with capsaicin did not cause sensory or motor blockade at the given concentrations. DT-A002 alone also did not cause sensory or motor blockade, but when combined with capsaicin, both sensory and motor blockade occurred, and the blockade was concentration-dependent. Ultimately, when capsaicin was at 1.73 mM and DT-A002 was at 1.4 mM, a good separation between sensory and motor blockade was achieved. Figure 12 As shown in the H&E and TB sections, no muscle inflammation or nerve damage was observed in any group, indicating that the biocompatibility remained good even when combined with capsaicin.

[0042] Example 6 Based on the beneficial results of DT-A002, this embodiment evaluates the systemic toxicity of compound DT-A001 to verify the potential for clinical translation of this class of compounds. Kunming mice were randomly divided into 5 groups of 6 mice each: a saline group and experimental groups receiving 10, 15, 20, and 25 mg / kg of DT-A002 solution, respectively. Before administration, the body weight of the Kunming mice was measured. 200 μL of DT-A002 was injected via the tail vein. The mice's condition was observed after administration, and their body weight was measured daily, with the number of deaths recorded. The observation period lasted for 14 consecutive days. Heart, liver, spleen, lung, kidney, and brain tissues were obtained through dissection. Organ coefficients were calculated using the formula "organ weight / body weight," and each organ was then subjected to H&E staining.

[0043] Mice in the 20.0 mg / kg group experienced mild shortness of breath after administration, which resolved within 5 minutes; no abnormalities were observed in the other groups after administration. Figure 13 As shown, the four concentrations of DT-A002 had little effect on mouse body weight, with no significant difference in weight change between the 20.0 mg / kg group and the saline group. Figure 14 and Figure 15 It can be seen that there are no significant differences in organ coefficients and organ slices in the drug group and the saline group compared with the saline group, indicating that DT-A002 has low systemic toxicity.

[0044] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0045] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.

Claims

1. A compound having a local anesthetic function, characterized in that, The structural formula is: Wherein, n is 3, 5 or 7.

2. A process for the preparation of a compound having local anaesthetic properties, characterized in that, The method comprises the following steps: S1, lidocaine is mixed with 2-bromoethanol and heated to obtain compound 1; S2, dicarboxylic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride and a solvent are mixed to obtain a mixed solution; compound 1, a base and a solvent are mixed and dissolved, and then the mixed solution is added; after spin-drying, separation and purification, a compound with local anesthetic function is obtained.

3. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The synthesis path of the compound with local anesthetic function is: 。 4. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The structural formula of the compound 1 is: 。 5. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The solvent is tetrahydrofuran or dichloromethane.

6. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The base is any one of DMAP, DIPEA, pyridine, triethylamine and Na2CO3.

7. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The dicarboxylic acid is any one of glutaric acid, heptanedioic acid and azelaic acid.

8. The method for preparing the compound with local anesthetic function according to claim 2, characterized in that, The molar ratio between the dicarboxylic acid, N-hydroxysuccinimide, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, compound 1 and the base is 1:2.4:2.4:2.4:2.

4.

9. Use of the compound of claim 1 in the preparation of an anesthetic or analgesic drug.

10. A medicament, characterized by comprising: The capsaicin and the compound of claim 1.

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