Lidocaine hydrochloride loaded polymer material as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicine synthesis, and particularly relates to a lidocaine hydrochloride loaded polymer material as well as a preparation method and application thereof. The lidocaine hydrochloride loaded polymer material provided by the invention comprises a carrier and lidocaine hydrochloride loaded in the carrier, the carrier is polyethyleneimine and polyacrylic acid which are physically crosslinked. According to the invention, lidocaine hydrochloride is loaded in physically crosslinked polyethyleneimine and polyacrylic acid, and the lidocaine hydrochloride can be directly applied to a part, can absorb moisture and can rapidly form gel; the data of the embodiment shows that the polymer material loaded with lidocaine hydrochloride can obviously prolong the analgesic time efficiency, and also can promote cell apoptosis and inhibit tumor growth by enhancing infiltration of CD8 + cells.

Description

Technical Field

[0001] The invention belongs to the technical field of drug synthesis, and particularly relates to a lidocaine hydrochloride-loaded polymer material, a preparation method thereof, and an application thereof. Background Art

[0002] Acute post-surgical pain (APSP) occurs after surgery-related tissue damage and is one of the common complications after surgery. Uncontrolled APSP may further develop into chronic post-operative pain, further increasing the medical burden.

[0003] The most commonly used analgesic medication for postoperative pain management is opioids, and opioids are the most widely used drugs. However, their high-dose use can lead to many related side effects, such as respiratory depression, nausea, vomiting, itching, intestinal dysfunction, and fatal overdose due to abuse. Therefore, postoperative pain management based on local anesthetics (LA) is a current research hotspot, mainly including continuous epidural analgesia, regional nerve block, and local wound infiltration. Compared with traditional opioids, postoperative analgesia based on LA has the advantages of lower price and fewer side effects, but LA has a short duration of action, usually no more than 8 hours, and overdose can still cause irritability, convulsions, and even life-threatening LA poisoning reactions.

[0004] To address the short duration of LA action and the potential for toxicity caused by overdose, current research focuses on developing various engineered LAs. These approaches primarily involve loading commonly used LAs (such as lidocaine hydrochloride, ropivacaine, and bupivacaine) onto various carriers. Common drug carriers include liposomes and graphene. However, liposomes can exhibit a burst release effect, resulting in unstable sustained-release effects. Graphene also presents similar issues and suffers from poor biocompatibility. Summary of the Invention

[0005] The purpose of the present invention is to provide a lidocaine hydrochloride loaded polymer material and a preparation method and application thereof. The lidocaine hydrochloride loaded polymer material provided by the present invention has good biocompatibility and sustained-release effect.

[0006] In order to achieve the above object, the present invention provides the following technical solutions:

[0007] The present invention provides a lidocaine hydrochloride loaded polymer material, comprising a carrier and lidocaine hydrochloride loaded in the carrier; the carrier is physically cross-linked polyethyleneimine and polyacrylic acid;

[0008] The mass percentage of lidocaine hydrochloride in the lidocaine hydrochloride loaded polymer material is 6 to 12%;

[0009] The porosity of the lidocaine hydrochloride loaded polymer material is 70%; the mass ratio of the polyethyleneimine to the polyacrylic acid is 1:1.

[0010] Preferably, the weight average molecular weight of the polyethyleneimine is 70,000; the weight average molecular weight of the polyacrylic acid is 240,000.

[0011] Preferably, the particle size of the lidocaine hydrochloride loaded polymer material is 100-400 μm.

[0012] Preferably, the pore size of the lidocaine hydrochloride loaded polymer material is 15 to 30 μm.

[0013] The present invention also provides a method for preparing the lidocaine hydrochloride loaded polymer material described in the above technical solution, comprising the following steps:

[0014] mixing lidocaine hydrochloride, a polyethyleneimine solution, and a polyacrylic acid solution to obtain a polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride;

[0015] The polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride is freeze-dried to obtain the lidocaine hydrochloride loaded polymer material.

[0016] Preferably, the freeze-drying comprises pre-freezing and vacuum drying performed sequentially; the pre-freezing temperature is -80°C; the time is 40 to 50 hours; the vacuum degree of the vacuum drying is ≤10Pa, and the time is 40 to 50 hours.

[0017] Preferably, the mass concentration of the polyethyleneimine solution is 10% (w / v); the mass concentration of the polyacrylic acid solution is 10% (w / v); and the mass concentration of lidocaine hydrochloride in the polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride is 6% (w / v).

[0018] Preferably, the volume ratio of the polyethyleneimine solution to the polyacrylic acid solution is 1:1.

[0019] The present invention also provides the lidocaine hydrochloride loaded polymer material described in the above technical solution or the use of the lidocaine hydrochloride loaded polymer material described in the above technical solution in the preparation of drugs with prolonged analgesic effect, anti-inflammatory drugs or anti-tumor drugs.

[0020] The present invention provides a lidocaine hydrochloride-loaded polymer material, comprising a carrier and lidocaine hydrochloride loaded in the carrier; the carrier is physically cross-linked polyethyleneimine and polyacrylic acid; the weight percentage of lidocaine hydrochloride in the lidocaine hydrochloride-loaded polymer material is 6-12%; the porosity of the lidocaine hydrochloride-loaded polymer material is 70%; and the mass ratio of the polyethyleneimine to the polyacrylic acid is 1:1. The present invention loads lidocaine hydrochloride in the physically cross-linked polyethyleneimine and polyacrylic acid. The material can be directly applied to the affected area, absorbs water, and rapidly forms a hydrogel. The hydrogel has a stable structure, thereby achieving a stable sustained release effect. Furthermore, the polyethyleneimine and polyacrylic acid are non-toxic and have good biocompatibility.

[0021] The data in the examples show that the polymer material loaded with lidocaine hydrochloride can significantly prolong the analgesic effect, and may also promote cell apoptosis and inhibit tumor growth by enhancing the infiltration of CD8+ cells.

[0022] Furthermore, the preparation method of the lidocaine hydrochloride loaded polymer material provided by the present invention is simple and easy to operate, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0024] Figure 1 This is a scanning electron micrograph of the LDC-PEI / PAA hydrogel prepared in Example 1;

[0025] Figure 2 This is a diagram showing the in vitro drug release effect of the LDC-PEI / PAA hydrogel prepared in Example 1;

[0026] Figure 3 Storage modulus (G') and loss modulus (G") plots for the LDC-PEI / PAA hydrogel prepared in Example 1;

[0027] Figure 4 The infrared spectrum analysis and XRD detection pattern of the LDC-PEI / PAA hydrogel prepared in Example 1;

[0028] Figure 5 This is the MWT of each group of mice in Application Example 1. DETAILED DESCRIPTION

[0029] The present invention provides a lidocaine hydrochloride loaded polymer material, comprising a carrier and lidocaine hydrochloride loaded in the carrier; the carrier is physically cross-linked polyethyleneimine and polyacrylic acid;

[0030] The mass percentage of lidocaine hydrochloride in the lidocaine hydrochloride loaded polymer material is 6-12%; the porosity of the lidocaine hydrochloride loaded polymer material is 70%; and the mass ratio of the polyethyleneimine to the polyacrylic acid is 1:1.

[0031] The present invention sets the loading amount of lidocaine hydrochloride within the above range, which can ensure that the lidocaine hydrochloride has a perfect effect and does not produce a poisoning reaction.

[0032] The lidocaine hydrochloride loaded polymer material provided by the present invention can directly act on the injured part and quickly (within 2 seconds) form a hydrogel state.

[0033] In one embodiment of the present invention, the weight-average molecular weight of the polyethyleneimine is 70,000; the weight-average molecular weight of the polyacrylic acid is 240,000. In another embodiment of the present invention, the mass ratio of the polyethyleneimine to the polyacrylic acid is 1:1. In another embodiment of the present invention, the percentage of lidocaine hydrochloride loaded by mass in the lidocaine hydrochloride-loaded polymer material is 12%.

[0034] As an embodiment of the present invention, the porosity of the lidocaine hydrochloride loaded polymer material is 70%.

[0035] The present invention provides a method for preparing a lidocaine hydrochloride loaded polymer material, comprising the following steps:

[0036] mixing lidocaine hydrochloride, polyethyleneimine (PEI) solution and polyacrylic acid (PAA) solution to obtain polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride;

[0037] The polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride is freeze-dried to obtain the lidocaine hydrochloride loaded polymer material.

[0038] As one embodiment of the present invention, the mass concentration of the polyethyleneimine solution can be 10% (w / v); the mass concentration of the polyacrylic acid solution can be 10% (w / v). As one embodiment of the present invention, the content of lidocaine hydrochloride in the polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride can be 6% (w / v).

[0039] As an embodiment of the present invention, the mixing is carried out under the condition of magnetic stirring.

[0040] As an embodiment of the present invention, the polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride has a porous network structure, which wraps the lidocaine hydrochloride therein.

[0041] As an embodiment of the present invention, the freeze-drying includes pre-freezing and vacuum drying performed in sequence; the pre-freezing temperature is -70 to -90°C, specifically -80°C; the time can be 40 to 50 hours, specifically 48 hours; the vacuum degree of the vacuum drying can be ≤10Pa, specifically 1Pa; the time can be 40 to 50 hours, specifically 48 hours.

[0042] The present invention also provides the use of the lidocaine hydrochloride loaded polymer material described in the above technical solution or the lidocaine hydrochloride loaded polymer material described in the above preparation method in the preparation of drugs with prolonged analgesic effect, anti-inflammatory drugs or anti-tumor drugs.

[0043] In order to further illustrate the present invention, the scheme of the present invention is described in detail below with reference to the accompanying drawings and embodiments, but they should not be understood as limiting the scope of protection of the present invention.

[0044] Sources of materials and parameters involved in the examples:

[0045] Polyethyleneimine MW70000; Shanghai MacLean Biochemical Technology Co., Ltd.;

[0046] Polyacrylic acid MW240000; Shanghai MacLean Biochemical Technology Co., Ltd.;

[0047] Lidocaine hydrochloride; Shanghai Aladdin Biochemical Technology Co., Ltd.

[0048] Example 1

[0049] LDC was dissolved in a 10% wt PAA solution to obtain a PAA solution containing 12 wt% LDC. The PAA solution containing 12 wt% LDC was mixed with a 10% (w / v) PEI solution in a 5:5 ratio of equal volumes to obtain a 6% (w / v) LDC-PEI / PAA hydrogel loaded with LDC (denoted as LDC-PEI / PAA hydrogel).

[0050] The LDC-loaded PEI / PAA hydrogel was placed in a -80°C refrigerator (pre-cooled for 48 h) in the dark, then vacuum-dried (vacuum drying at a vacuum degree of 1 Pa for 48 h) and ground to obtain a lidocaine hydrochloride-loaded polymer material (the mass percentage of lidocaine hydrochloride was 11.4%).

[0051] The particle size of the lidocaine hydrochloride loaded polymer material obtained in Example 1 is 300 μm; the pore size is 15 μm, and the mass percentage of lidocaine hydrochloride in the lidocaine hydrochloride loaded polymer material is 11.4%.

[0052] Example 2

[0053] The only difference from Example 1 is that a PAA solution containing 12 wt % LDC was mixed with a PEI solution in a volume ratio of 6:4 to obtain a lidocaine hydrochloride-loaded polymer material with a particle size of 200 μm (the mass percentage of lidocaine hydrochloride was 11.4%) and a pore size of 20 μm.

[0054] Example 3

[0055] The only difference from Example 1 is that a PAA solution containing 12 wt % LDC was mixed with a PEI solution in a volume ratio of 4:6 to obtain a lidocaine hydrochloride-loaded polymer material with a particle size of 250 μm (the mass percentage of lidocaine hydrochloride was 11.4%) and a pore size of 15 μm.

[0056] Figure 1 This is a scanning electron micrograph of the LDC-PEI / PAA hydrogel prepared in Example 1. Figure 1 It can be seen that the LDC-PEI / PAA hydrogel prepared in Example 1 has a porous network structure.

[0057] An in vitro drug release experiment was conducted on the LDC-PEI / PAA hydrogel prepared in Example 1. The experimental steps were as follows: simulating the in vivo environmental temperature, the LDC-PEI / PAA hydrogel powder was placed in a centrifuge tube to form a hydrogel at 37°C, and 1 mL was taken; 1% phosphate buffer solution (1% PBS, 4 mL, 37°C) was added to the centrifuge tube and placed in a constant temperature oscillating box at 37°C and 70 rpm. At the specified time (2nd, 4th, 6th, 8th, 10th, 12th, 24th, 48th, 72th, 96th, 120h), 1 mL of solution was collected, and after collecting the solution, 1 mL of 37°C 1% PBS solution was added to the centrifuge tube. Finally, the concentration of lidocaine hydrochloride was determined by testing the absorbance at 263 nm using a UV-visible spectrophotometer, and the drug release curve was drawn. The experimental results are shown in Figure 2 ,In vitro drug release results showed that LDC released about 60% at 12 h, ,and about 75% at 24 h, and the total release of LDC exceeded 120 h.

[0058] The storage modulus (G') and loss modulus (G") of the LDC-PEI / PAA hydrogel prepared in Example 1 were tested using a German Haake Mars40 rheometer. The test results are shown in Figure 3 ,from Figure 3It can be seen that the G' of the LDC-PEI / PAA hydrogel prepared in Example 1 is always greater than G", and its G' and G" always change with frequency, indicating that the LDC-PEI / PAA hydrogel is formed by physical cross-linking and has strong tissue adhesion.

[0059] The LDC-PEI / PAA hydrogel of Example 1 was subjected to infrared spectroscopy and XRD detection, and the test results are shown in FIG. Figure 4 A and B, from Figure 4 From A, we can see that LDC is at 3300cm -1 and 1600cm -1 The characteristic peak of amide group (-NH-CO-) is shown near 3000 cm -1 The characteristic peak of aromatic ring (CH) is shown near PEI / PAA at 1600 cm -1 The characteristic peaks of amino (-NH) and carboxyl (-COOH) are shown near the surface, where amino is the functional group of PEI and carboxyl is the functional group of PAA; LDC-PEI / PAA is at 3300 cm -1 and 1600cm -1 The characteristic peak of amide group (-NH-CO-) is shown near 3000 cm -1 The characteristic peak of aromatic ring (CH) is shown near 1600 cm -1 The characteristic peaks of amino (-NH) and carboxyl (-COOH) groups were shown nearby. The results showed that PEI and PAA formed PEI / PAA hydrogel through physical crosslinking, and LDC was successfully loaded on the hydrogel. Figure 4 Figure B also verifies the successful synthesis of the above-mentioned LDC-PEI / PAA hydrogel.

[0060] Application Examples

[0061] Application Example 1: Study on the analgesic time effect in the mouse plantar incision model

[0062] (1) Animal grouping

[0063] Thirty BALB / c mice were randomly divided into five groups using a random number table, with six mice in each group: blank control group (Comparative Example 1: Blank group), model group (Comparative Example 2: Ctrl group), simple LDC group (Comparative Example 3: LDC group), simple PEI / PAA hydrogel group (Comparative Example 4: PEI / PAA group), and LDC-PEI / PAA hydrogel group (Comparative Example 5: LDC-PEI / PAA group).

[0064] (2) Model establishment

[0065] The model was established under general anesthesia and aseptic conditions. To prevent the mice from dying from hypothermia after anesthesia, all operations were performed on a heating blanket.

[0066] (3) Mechanical Withdrawal Threshold (MWT) measurement

[0067] The basal MWT of each group of mice should be measured before modeling, recorded as base, and the MWT of each group of mice should be measured once at 0, 4, 8, 12, 24, 48, 72, 96, and 120 hours after modeling (measurement results are shown in Figure 5 , Figure 5 Figure A is a schematic diagram of the mouse plantar incision model, and Figure B is the MWT of each group of mice. 30 minutes before measurement, mice were placed on a wire mesh to isolate each other and familiarize themselves with the environment. VonFrey fibers were then used to stimulate the soles of their left feet from under the wire mesh to observe their reactions. The weights of the VonFrey fibers were 0.008, 0.02, 0.04, 0.07, 0.16, 0.40, 0.60, 1.00, and 1.40 g, respectively. First, 0.40 g of fiber was used to stimulate the soles of the mice's feet. If the mice showed an avoidance reaction, such as withdrawing their legs, it was recorded as "×" and a lighter fiber was used. If the mice showed no obvious reaction, it was recorded as "0" and a heavier fiber was used. This cycle was repeated 6 times, with a 5-minute interval between each time. The weight of the fiber used last time was recorded to obtain a set of data consisting of six "×"s and "0s". The final result was obtained through processing, which was the MWT of the mice. After all cycle measurements are completed, the experimental data are analyzed by drawing a line graph.

[0068] Results: The MWT of the LDC-PEI / PAA group in comparative example 5 was significantly increased, which was consistent with the results of the in vitro drug sustained release experiment. The LDC-PEI / PAA hydrogel significantly prolonged the analgesic effect of LDC.

[0069] Application Example 2: Comparison of inflammatory factor levels in each group of mice

[0070] In each comparative example of Application Example 1, blood samples were collected from the retroorbital venous plexus of mice 24 hours after modeling, and the expression levels of inflammatory factors IL-1β, IL-6, and TNF-α were observed using Masson staining sections after ELISA testing, further demonstrating the biocompatibility of LDC-PEI / PAA hydrogel.

[0071] Results: The expression levels of inflammatory factors IL-1β, IL-6, and TNF-α in the LDC-PEI / PAA group in Comparative Example 5 were significantly reduced, indicating that the LDC-PEI / PAA hydrogel has good biocompatibility and anti-inflammatory potential.

[0072] Application Example 3: Pathological changes in the plantar surface of mice in each group

[0073] In each comparative example of Application Example 1, plantar specimens were collected on the 7th and 14th day after modeling and HE staining was performed to analyze the plantar inflammation.

[0074] Results: The inflammatory infiltration of the LDC-PEI / PAA group in Example 5 was significantly reduced on the 7th and 14th days after modeling, which verified the conclusion of Application Example 2.

[0075] Application Example 4: Effect of LDC-PEI / PAA Hydrogel on Tumor-Bearing Mice

[0076] (1) Animal grouping

[0077] Thirty BALB / c mice were randomly divided into five groups using a random number table, with six mice in each group: model group (Comparative Example 1: Uncut group), unresected irradiation group (Comparative Example 2: Uncut+RT group), simple LDC-PEI / PAA hydrogel group (Comparative Example 3: Cut+Gel group), resected irradiation group (Comparative Example 4: Cut+RT group), and irradiated LDC-PEI / PAA hydrogel group (Comparative Example 5: Cut+RT+Gel group).

[0078] (2) Model establishment

[0079] The model was established under general anesthesia and aseptic conditions. To prevent the mice from dying from hypothermia after anesthesia, all operations were performed on a heating blanket.

[0080] (3) The mice in each comparison example were treated accordingly on the 7th day after inoculation: Comparative Example 1 received no treatment; Comparative Example 2 was given radiation therapy without tumor removal; Comparative Example 3 was given LDC-PEI / PAA hydrogel after tumor removal; Comparative Example 4 was given radiation therapy after tumor removal; Comparative Example 5 was given radiation therapy and LDC-PEI / PAA hydrogel after tumor removal.

[0081] (4) The MWT, body weight, tumor volume and paralysis time of mice in each comparison example were measured.

[0082] (4) Another set of experiments was conducted using the same method. Tumor specimens from each comparison group of mice were collected 1 week after treatment for Tunel staining to observe cell apoptosis, and liver and tumor specimens were collected for flow cytometry analysis.

[0083] Results: Mice in the Cut+RT+Gel group (Comparative Example 5) showed a significant increase in MWT, demonstrating a favorable analgesic effect. Tumor volume growth was the slowest, demonstrating anti-tumor potential, and paralysis was the longest, demonstrating its favorable tumor growth inhibition effect. Tunel staining revealed that mice in the Cut+RT+Gel group (Comparative Example 5) showed the most tumor apoptosis, and flow cytometry revealed the highest CD8+ cell (cytotoxic T lymphocyte) infiltration. These results suggest that its anti-tumor effect may be achieved by increasing CD8+ cell infiltration and thereby promoting tumor cell apoptosis.

[0084] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. Other embodiments can be obtained based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A lidocaine hydrochloride-loaded polymer material, comprising a carrier and lidocaine hydrochloride loaded in the carrier; the carrier is physically cross-linked polyethyleneimine and polyacrylic acid; The mass percentage of lidocaine hydrochloride in the lidocaine hydrochloride loaded polymer material is 6 to 12%; The porosity of the lidocaine hydrochloride loaded polymer material is 70%; the mass ratio of the polyethyleneimine to the polyacrylic acid is 1:

1.

2. The lidocaine hydrochloride loaded polymer material according to claim 1, wherein The weight average molecular weight of the polyethyleneimine is 70,000; the weight average molecular weight of the polyacrylic acid is 240,000.

3. The lidocaine hydrochloride loaded polymer material according to claim 1 or 2, characterized in that: The particle size of the lidocaine hydrochloride loaded polymer material is 100-400 μm.

4. The lidocaine hydrochloride loaded polymer material according to claim 1 or 2, characterized in that: The pore size of the lidocaine hydrochloride loaded polymer material is 15 to 30 μm.

5. A method for preparing the lidocaine hydrochloride loaded polymer material according to any one of claims 1 to 4, comprising the following steps: mixing lidocaine hydrochloride, a polyethyleneimine solution, and a polyacrylic acid solution to obtain a polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride; The polyethyleneimine / polyacrylic acid hydrogel loaded with lidocaine hydrochloride is freeze-dried to obtain the lidocaine hydrochloride loaded polymer material.

6. The preparation method according to claim 5, wherein The freeze drying comprises pre-freezing and vacuum drying carried out in sequence; the pre-freezing temperature is -80°C; the time is 40 to 50 hours; the vacuum degree of the vacuum drying is ≤10Pa, and the time is 40 to 50 hours.

7. The preparation method according to claim 5, wherein The mass concentration of the polyethyleneimine solution is 10% (w / v); the mass concentration of the polyacrylic acid solution is 10% (w / v); The mass concentration of lidocaine hydrochloride in the lidocaine hydrochloride-loaded polyethyleneimine / polyacrylic acid hydrogel is 6% (w / v).

8. The preparation method according to claim 5, wherein The volume ratio of the polyethyleneimine solution to the polyacrylic acid solution is 1:

1.

9. Use of the lidocaine hydrochloride-loaded polymer material according to any one of claims 1 to 4 or the lidocaine hydrochloride-loaded polymer material according to any one of claims 5 to 8 in the preparation of drugs with prolonged analgesic effect, anti-inflammatory drugs or anti-tumor drugs.

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