Lignin-carbohydrate complex anti-intestinal cancer drug delivery material capable of promoting drug to enter cells and preparation method of lignin-carbohydrate complex anti-intestinal cancer drug delivery material

Abstract

The invention provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material capable of promoting drug entry and a preparation method of the lignin-carbohydrate complex anti-intestinal cancer drug delivery material, and belongs to the field of biomedical materials. The preparation method comprises the following steps: (1) dissolving a lignin-carbohydrate complex in water, and performing ultrasonic-assisted mixing with an ethanol solution of an anti-intestinal cancer drug at room temperature to obtain a mixed solution; (2) adding salt into the mixed solution obtained in the step (1), and stirring at room temperature to obtain a solution; and (3) transferring the solution obtained in the step (2) into a dialysis bag, dialyzing in water, and freeze-drying or vacuum-drying the dialysate to obtain the lignin-carbohydrate complex anti-intestinal cancer drug delivery material capable of promoting the drug to enter cells. The delivery material prepared by the method can effectively protect the activity of the anti-intestinal cancer drug, prevent the anti-intestinal cancer drug from being decomposed or losing efficacy in the stomach and the intestinal tract and improve the drug delivery effect, and the method is simple and low in cost.

Description

Technical Field

[0001] The present invention relates to the field of biomedical materials, and in particular to a lignin-carbohydrate complex anti-intestinal cancer drug delivery material capable of promoting drug entry into cells and a preparation method thereof. Background Art

[0002] Intestinal cancer, also known as large intestine cancer or colorectal cancer, is a malignant tumor that occurs in the colon or rectum. Common treatment methods include surgery, radiotherapy, chemotherapy, etc., but these methods are relatively traumatic or have side effects on patients. Although oral anticancer drugs can reduce physical trauma, the drugs used, such as 5-fluorouracil (5-FU), irinotecan, oxaliplatin, etc., often bring many adverse reactions to patients, and cannot reach the diseased cells accurately and efficiently, resulting in unsatisfactory efficacy and therapeutic effects. The main reasons are as follows: First, most drugs are inactivated or decomposed in the stomach and cannot reach the intestinal lesions completely; second, even if they reach the intestines, they are decomposed by the metabolism of many intestinal microorganisms, making the drugs ineffective; third, even if they reach the lesion site, they are blocked by the cell membrane and enter the lesion cells at a low rate, so they are excreted. Summary of the invention

[0003] The present invention provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material capable of promoting drug entry into cells and a preparation method thereof. The anti-intestinal cancer drug delivery material prepared by the present invention can accurately and efficiently reach the lesion site.

[0004] The present invention provides a method for preparing a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells, comprising the following steps:

[0005] (1) dissolving the lignin-carbohydrate complex in water and then mixing it with an ethanol solution of an anti-colon cancer drug under ultrasonic assistance at room temperature to obtain a mixed solution;

[0006] (2) adding salt to the mixed solution obtained in step (1), stirring at room temperature to obtain a solution;

[0007] (3) The solution obtained in step (2) is transferred to a dialysis bag and dialyzed in water, and the dialysate is freeze-dried or vacuum-dried to obtain the lignin-carbohydrate complex anti-colon cancer drug delivery material that can promote drug entry into cells.

[0008] Preferably, in step (1), the ratio of the mass of the lignin-carbohydrate complex to the volume of water is 1:10 to 1:1000, the unit of the mass is g, and the unit of the volume is mL;

[0009] The lignin-carbohydrate complex is Any one of lignin-carbohydrate complex, enzymatic lignin-carbohydrate complex, organic solvent lignin-carbohydrate complex, ionic liquid lignin-carbohydrate complex, and deep eutectic solvent lignin-carbohydrate complex;

[0010] The carbohydrate mass content in the lignin-carbohydrate complex is 5-30%.

[0011] Preferably, in step (1), the mass concentration of the anti-colon cancer drug in the ethanol solution of the anti-colon cancer drug is 0.1-30%.

[0012] Preferably, the anti-colon cancer drug is any one of paclitaxel, camptothecin, vincristine, resveratrol, quercetin, bryostatin, tanshinone IIA, cyclophosphamide, gemcitabine, cytarabine, carboplatin, oxaliplatin, cisplatin, etoposide, teniposide, pemetrexed, 5-fluorouracil and imatinib.

[0013] Preferably, in step (1), the volume ratio of water to the ethanol solution of the anti-colon cancer drug is 10:1 to 100:1.

[0014] Preferably, in step (1), the power of the ultrasound during the ultrasound-assisted mixing is 100 to 1000 W, and the time of the ultrasound-assisted mixing is 1 to 30 min.

[0015] Preferably, in step (2), the salt is any one of NaCl, KCl, and CaCl2;

[0016] The volume ratio of the mass of the salt to the water in step (1) is 0.0001:1 to 0.001:1, the unit of the mass is g, and the unit of the volume is mL;

[0017] Preferably, in step (2), the stirring speed is 100 to 1000 rpm and the time is 1 to 10 min.

[0018] Preferably, in step (3), the molecular weight cutoff of the dialysis bag is 500 to 1000 Da, and the dialysis time is 0.5 to 2 h;

[0019] The freeze drying temperature is -50 to -10°C, the pressure is 10 to 500 Pa, and the time is 6 to 24 hours; the vacuum drying temperature is 30 to 60°C, the pressure is -0.02 to -0.1 MPa, and the time is 6 to 12 hours.

[0020] The present invention also provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that is prepared by the preparation method described in the above technical solution and can promote drug entry into cells.

[0021] Lignin-carbohydrate complex is a natural polymer complex in which lignin and carbohydrates (generally hemicellulose) in plant cell walls are connected by ether bonds or ester bonds. It has a unique hydrophobic (lignin end) and hydrophilic (hemicellulose end) structure, and is an ideal anti-intestinal cancer drug delivery material; the anti-intestinal cancer drug is hydrophobic and can be combined with the lignin end, while the hydrophilic end of the lignin-carbohydrate complex is at the periphery, and a unique shell-core structure is formed by this self-assembly method, and the anti-intestinal cancer drug is at the core position. The lignin-carbohydrate complex coats the anti-intestinal cancer drug to form a shell, which effectively protects the anti-intestinal cancer drug, thereby preventing it from decomposing or inactivating in the stomach and intestines, and the polysaccharide in the shell has acid and microbial responsiveness, and is subject to the dissolution mechanism of gastric acid and microorganisms in the stomach and intestines, so that the drug can diffuse from the shell in the intestines to act on the lesion position. Therefore, the anti-intestinal cancer drug delivery material of the present invention can effectively protect the activity of the anti-intestinal cancer drug, avoid its decomposition or failure in the stomach and intestines, thereby improving its drug delivery effect.

[0022] The core-shell drug delivery material has a large number of negatively charged groups (-OH, -COOH, etc.) on its surface, which can load metal cations such as Na + , K + , Ca 2+ , these ions are easily recognized by the cell membrane and can pass through the cell membrane efficiently.

[0023] The preparation method of the anti-intestinal cancer drug delivery material of the present invention is simple and low in cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is an atomic force micrograph of the lignin-carbohydrate complex anti-colon cancer drug delivery material of Example 1. DETAILED DESCRIPTION

[0025] The present invention provides a method for preparing a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells, comprising the following steps:

[0026] (1) dissolving the lignin-carbohydrate complex in water and then mixing it with an ethanol solution of an anti-colon cancer drug under ultrasonic assistance at room temperature to obtain a mixed solution;

[0027] (2) adding salt to the mixed solution obtained in step (1), stirring at room temperature to obtain a solution;

[0028] (3) The solution obtained in step (2) is transferred to a dialysis bag and dialyzed in water, and the dialysate is freeze-dried or vacuum-dried to obtain the lignin-carbohydrate complex anti-colon cancer drug delivery material that can promote drug entry into cells.

[0029] The present invention dissolves the lignin-carbohydrate complex in water and then mixes it with an ethanol solution of an anti-intestinal cancer drug under ultrasonic assistance at room temperature to obtain a mixed solution.

[0030] In the present invention, the ultrasonic power during the ultrasonic-assisted mixing is preferably 100-1000 W, and the time of the ultrasonic-assisted mixing is preferably 1-30 min. In a specific embodiment of the present invention, the ultrasonic power can be 100 W, 200 W, 300 W, 400 W, 500 W, 600 W, 700 W, 800 W, 900 W or 1000 W, and the time of the ultrasonic-assisted mixing can be 1 min, 5 min, 10 min, 15 min, 20 min, 25 min or 30 min.

[0031] In the present invention, the mass ratio of the lignin-carbohydrate complex to the volume ratio of water is preferably 1:10 to 1:1000, the unit of the mass is g, and the unit of the volume is mL. In a specific embodiment of the present invention, the mass ratio of the lignin-carbohydrate complex to water can be 1:10 to 1000, 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900 or 1:1000.

[0032] In the present invention, the lignin-carbohydrate complex preferably comprises One or more of lignin-carbohydrate complexes, enzymatic lignin-carbohydrate complexes, organic solvent lignin-carbohydrate complexes, ionic liquid lignin-carbohydrate complexes, and deep eutectic solvent lignin-carbohydrate complexes.

[0033] In the present invention, the mass content of carbohydrates in the lignin-carbohydrate complex is preferably 5-30%. In a specific embodiment of the present invention, the mass content of carbohydrates in the lignin-carbohydrate complex can be 5%, 10%, 15%, 20%, 25% or 30%.

[0034] In the present invention, the volume ratio of water to the ethanol solution of the anti-intestinal cancer drug is preferably 10 to 100:1; in a specific embodiment of the present invention, the volume ratio of water to the ethanol solution of the anti-intestinal cancer drug can be 10:1, 20:1, 30:1, 40:1, 50:1, 60:1, 70:1, 80:1, 90:1 or 100:1;

[0035] In the present invention, the mass concentration of the anti-colon cancer drug in the ethanol solution of the anti-colon cancer drug is preferably 0.1-30%. In a specific embodiment of the present invention, the mass concentration of the anti-colon cancer drug in the ethanol solution of the anti-colon cancer drug can be 0.1%, 1%, 5%, 10%, 15%, 20%, 25% or 30%.

[0036] In the present invention, the anti-colon cancer drug is preferably any one of paclitaxel, camptothecin, vincristine, resveratrol, quercetin, bryostatin, tanshinone IIA, cyclophosphamide, gemcitabine, cytarabine, carboplatin, oxaliplatin, cisplatin, etoposide, teniposide, pemetrexed, 5-fluorouracil and imatinib.

[0037] After obtaining the mixed solution, the present invention adds salt to the mixed solution obtained in step (1), and stirs at room temperature to obtain a solution.

[0038] In the present invention, the stirring speed is preferably 100-1000rpm, and the time is preferably 1-10min. In a specific embodiment of the present invention, the stirring speed may be 100rpm, 200rpm, 300rpm, 400rpm, 500rpm, 600rpm, 700rpm, 800rpm, 900rpm or 1000rpm, and the time may be 1min, 2min, 3min, 4min, 5min, 6min, 7min, 8min, 9min or 10min.

[0039] In the present invention, the volume ratio of the mass of the salt to the water in step (1) is preferably 0.0001:1 to 0.001:1, the unit of the mass is g, and the unit of the volume is mL. In a specific embodiment of the present invention, the volume ratio of the mass of the salt to the water in step (1) can be 0.0001:1, 0.0002:1, 0.0003:1, 0.0004:1, 0.0005:1, 0.0006:1, 0.0007:1, 0.0008:1, 0.0009:1 or 0.001:1; the salt is preferably any one of NaCl, KCl, and CaCl2.

[0040] After obtaining the solution, the present invention transfers the solution obtained in step (2) into a dialysis bag and dialyzes it in water, and then freeze-dries or vacuum-dries the dialysate to obtain the lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells.

[0041] In the present invention, the molecular weight cutoff of the dialysis bag is preferably 500-1000Da. In a specific embodiment of the present invention, the molecular weight cutoff of the dialysis bag may be 500Da, 600Da, 700Da, 800Da, 900Da or 1000Da.

[0042] In the present invention, the dialysis time is preferably 0.5 to 2 hours. In a specific embodiment of the present invention, the dialysis time can be 0.5 hours, 1 hour, 1.5 hours or 2 hours.

[0043] The purpose of dialysis is to remove the anticancer drugs and salt ions that are not bound to the lignin-carbohydrate complex and to improve the purity of the product.

[0044] In the present invention, the freeze-drying temperature is preferably -50 to -10°C, the pressure is preferably 10 to 500 Pa, and the time is preferably 6 to 24 h. In a specific embodiment of the present invention, the freeze-drying temperature may be -50°C, -40°C, -30°C, -20°C or -10°C, the pressure may be 10 Pa, 100 Pa, 200 Pa, 300 Pa, 400 Pa or 500 Pa, and the time may be 6 h, 12 h, 18 h or 24 h. The vacuum drying temperature is preferably 30 to 60°C, the pressure is preferably -0.02 to -0.1 MPa, and the time is preferably 6 to 12 h. In a specific embodiment of the present invention, the vacuum drying temperature may be 30°C, 40°C, 50°C or 60°C, the pressure may be -0.02 MPa, -0.04 MPa, -0.06 MPa, -0.08 MPa or -0.1 MPa, and the time may be 6 h, 8 h, 10 h or 12 h.

[0045] The present invention also provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that is prepared by the preparation method described in the above technical solution and can promote drug entry into cells.

[0046] The lignin-carbohydrate complex anti-colon cancer drug delivery material capable of promoting drug entry into cells and the preparation method thereof provided by the present invention are described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0047] In the embodiment: The preparation method of lignin-carbohydrate complexes can be found in the literature (DOI: 10.1186 / s13068-018-1262-1), the preparation method of enzymatic lignin-carbohydrate complexes can be found in the literature (DOI: 10.1515 / HF.2006.025), the preparation method of organic solvent lignin-carbohydrate complexes can be found in the literature (Peng et al., BioResources, 2014, 9(1): 1276-1289), the preparation method of ionic liquid lignin-carbohydrate complexes can be found in the literature (DOI: 10.3390 / molecules26195740), and the preparation method of low eutectic solvent lignin-carbohydrate complexes can be found in the literature (DOI: 10.1016 / j.carbpol.2023.121452).

[0048] Example 1

[0049] This embodiment provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells and a preparation method thereof, the steps are:

[0050] (1) 1g The lignin-carbohydrate complex (carbohydrate content of 5%) was dissolved in 10 mL of water and mixed with 1 mL of 0.1% paclitaxel ethanol solution at room temperature using 100 W ultrasonic wave assistance for 30 min;

[0051] (2) adding 0.001 g of NaCl to the mixed solution obtained in step (1), and stirring at room temperature and a rotation speed of 100 rpm for 10 min;

[0052] (3) The solution obtained in step (2) was transferred to a 500Da dialysis bag and dialyzed in water for 0.5 h. The dialysate was then freeze-dried at a temperature of -50°C and a pressure of 10 Pa for 6 h to obtain a lignin-carbohydrate complex anti-colon cancer drug delivery material with a drug loading of 46% and an encapsulation efficiency of 92%.

[0053] Using a rat orthotopic transplanted small intestinal tumor model, the drug was orally administered at a dose of 80 mg / kg per rat once every two days. After one month, the volume of the rat small intestinal tumor decreased by 20% and the weight decreased by 35%.

[0054] Figure 1 This is an atomic force micrograph of the lignin-carbohydrate complex anti-colon cancer drug delivery material of Example 1.

[0055] from Figure 1 The shell-core structure can be clearly observed in the experiment, which effectively protects the anticancer drugs in the core position and avoids decomposition or inactivation in the stomach and intestines.

[0056] Example 2

[0057] This embodiment provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells and a preparation method thereof, the steps are:

[0058] (1) 1 g of enzymatically hydrolyzed lignin-carbohydrate complex (carbohydrate content of 30%) was dissolved in 1000 mL of water and mixed with 10 mL of 30% camptothecin ethanol solution at room temperature using 1000 W ultrasonic wave for 1 min;

[0059] (2) adding 1 g of KCl to the mixed solution obtained in step (1), and stirring at room temperature and a rotation speed of 1000 rpm for 1 min;

[0060] (3) The solution obtained in step (2) was transferred to a 1000Da dialysis bag and dialyzed in water for 2 hours. The dialysate was then freeze-dried at a temperature of -10°C and a pressure of 500Pa for 24 hours to obtain a lignin-carbohydrate complex anti-colon cancer drug delivery material with a drug loading of 58% and an encapsulation efficiency of 70%.

[0061] AOM / DSS CRC colon cancer rat model was used. The drug was orally administered at a dose of 80 mg / kg per rat once every two days. After one month of administration, the volume of colon cancer in the rats decreased by 28% and the weight decreased by 37%.

[0062] Example 3

[0063] This embodiment provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells and a preparation method thereof, the steps are:

[0064] (1) 1 g of organic solvent lignin-carbohydrate complex (carbohydrate content of 10%) was dissolved in 100 mL of water and mixed with 5 mL of 10% bryostatin ethanol solution at room temperature using 500 W ultrasonic assistance for 10 min;

[0065] (2) adding 0.02 g of CaCl2 to the mixed solution obtained in step (1), and stirring at room temperature and a rotation speed of 500 rpm for 3 min;

[0066] (3) The solution obtained in step (2) was transferred to a 600Da dialysis bag and dialyzed in water for 1 hour. The dialysate was then freeze-dried at a temperature of -30°C and a pressure of 300Pa for 18 hours to obtain a lignin-carbohydrate complex anti-colon cancer drug delivery material with a drug loading of 45% and an encapsulation efficiency of 86%.

[0067] A rat orthotopic transplanted cecal tumor model was used, and the drug was orally administered at a dose of 80 mg / kg per rat once every two days. After one month, the volume of the rat cecal tumor decreased by 24% and the weight decreased by 32%.

[0068] Example 4

[0069] This embodiment provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells and a preparation method thereof, the steps are:

[0070] (1) 1 g of ionic liquid lignin-carbohydrate complex (carbohydrate content of 20%) was dissolved in 400 mL of water and mixed with 5 mL of 20% oxaliplatin ethanol solution at room temperature using 800 W ultrasonic wave-assisted mixing for 15 min;

[0071] (2) adding 0.2 g NaCl to the mixed solution obtained in step (1), and stirring at room temperature and a rotation speed of 700 rpm for 6 min;

[0072] (3) The solution obtained in step (2) was transferred to a 700Da dialysis bag and dialyzed in water for 1.5 hours. The dialysate was then vacuum dried at a temperature of 30°C and a pressure of -0.1MPa for 12 hours to obtain a lignin-carbohydrate complex anti-colon cancer drug delivery material with a drug loading of 53% and an encapsulation efficiency of 95%.

[0073] A rat colorectal cancer model was induced using dimethylbenzylhydrazine. The drug was orally administered at a dose of 80 mg / kg per rat once every two days. After one month, the volume of the rat colorectal cancer was reduced by 35% and the weight was reduced by 43%.

[0074] Example 5

[0075] This embodiment provides a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells and a preparation method thereof, the steps are:

[0076] (1) 1 g of a low eutectic solvent lignin-carbohydrate complex (carbohydrate content of 25%) was dissolved in 800 mL of water and mixed with 20 mL of 1% imatinib ethanol solution at room temperature using 400 W ultrasonic assistance for 20 min;

[0077] (2) adding 0.6 g KCl to the mixed solution obtained in step (1), and stirring at room temperature and a rotation speed of 400 rpm for 4 min;

[0078] (3) The solution obtained in step (2) was transferred to an 800Da dialysis bag and dialyzed in water for 1.2 hours. The dialysate was then vacuum dried at a temperature of 60°C and a pressure of -0.02MPa for 6 hours to obtain a lignin-carbohydrate complex anti-colon cancer drug delivery material with a drug loading of 55% and an encapsulation efficiency of 98%.

[0079] AOM / DSS CRC rectal cancer rat model was used, and the drug was orally administered at a dose of 80 mg / kg per rat once every two days. After one month, the volume of the rectal cancer in the rats decreased by 26% and the weight decreased by 30%.

[0080] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A method for preparing a lignin-carbohydrate complex anti-intestinal cancer drug delivery material that can promote drug entry into cells, characterized in that: The following steps are involved: (1) dissolving the lignin-carbohydrate complex in water and then mixing it with an ethanol solution of an anti-colon cancer drug under ultrasonic assistance at room temperature to obtain a mixed solution; (2) adding salt to the mixed solution obtained in step (1), stirring at room temperature to obtain a solution; (3) The solution obtained in step (2) is transferred to a dialysis bag and dialyzed in water, and the dialysate is freeze-dried or vacuum-dried to obtain the lignin-carbohydrate complex anti-colon cancer drug delivery material that can promote drug entry into cells.

2. The preparation method according to claim 1, characterized in that: In step (1), the ratio of the mass of the lignin-carbohydrate complex to the volume of water is 1:10 to 1:1000, the unit of the mass is g, and the unit of the volume is mL; The lignin-carbohydrate complex is Any one of lignin-carbohydrate complex, enzymatic lignin-carbohydrate complex, organic solvent lignin-carbohydrate complex, ionic liquid lignin-carbohydrate complex, and deep eutectic solvent lignin-carbohydrate complex; The carbohydrate mass content in the lignin-carbohydrate complex is 5-30%.

3. The preparation method according to claim 1, characterized in that: In step (1), the mass concentration of the anti-colon cancer drug in the ethanol solution of the anti-colon cancer drug is 0.1-30%.

4. The preparation method according to claim 1, characterized in that: The anti-colon cancer drug is any one of paclitaxel, camptothecin, vincristine, resveratrol, quercetin, bryostatin, tanshinone IIA, cyclophosphamide, gemcitabine, cytarabine, carboplatin, oxaliplatin, cisplatin, etoposide, teniposide, pemetrexed, 5-fluorouracil and imatinib.

5. The preparation method according to claim 1, characterized in that: In step (1), the volume ratio of water to the ethanol solution of the anti-colon cancer drug is 10:1 to 100:

1.

6. The preparation method according to claim 1, characterized in that: In step (1), the power of the ultrasonic wave during the ultrasonic wave-assisted mixing is 100 to 1000 W, and the time of the ultrasonic wave-assisted mixing is 1 to 30 minutes.

7. The preparation method according to claim 1, characterized in that: In step (2), the salt is any one of NaCl, KCl, and CaCl2; The mass ratio of the salt to the volume ratio of the water in step (1) is 0.0001:1 to 0.001:1, the unit of the mass is g, and the unit of the volume is mL.

8. The preparation method according to claim 1, characterized in that: In step (2), the stirring speed is 100 to 1000 rpm and the time is 1 to 10 min.

9. The preparation method according to claim 1, characterized in that: In step (3), the molecular weight cutoff of the dialysis bag is 500 to 1000 Da, and the dialysis time is 0.5 to 2 h; The freeze drying temperature is -50 to -10°C, the pressure is 10 to 500 Pa, and the time is 6 to 24 hours; the vacuum drying temperature is 30 to 60°C, the pressure is -0.02 to -0.1 MPa, and the time is 6 to 12 hours.

10. The lignin-carbohydrate complex anti-colon cancer drug delivery material capable of promoting drug entry into cells prepared by the preparation method according to any one of claims 1 to 9.

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