WO2022021508A1 - Method and system for using pulsed laser to control drug release - Google Patents

Abstract

A method and a system for using a pulsed laser to control drug release, relating to the technical field of pulsed laser applications. A solution (6) used in the invention comprises an agarose hydrogel (8) carrier, an anti-cancer drug (9), and a black phosphorus photosensitizer (10) loaded in the hydrogel (8) carrier. The anti-cancer drug (9) is encapsulated into the agarose hydrogel (8) loaded with the black phosphorus photosensitizer (10). A 780-110 nm near infrared pulsed laser provided in the system causes a photothermal reaction in the black phosphorus photosensitizer (10), accomplishing conversion of the hydrogel (8) from a gel state to a sol state, thereby implementing a process of release of the drug (9). The energy in single pulses of a pulsed laser can be controlled, and the irradiation space thereof is highly precise, and therefore the provided method and a system for using a pulsed laser to control the release of a drug (9) is able to precisely control the release position of the drug (9). By means of controlling the laser energy and pulse count of a pulsed laser, precise control of the amount of the drug (9) released is made possible, the invention thus having a high clinical value.

Description

A method and system for controlling drug release using pulsed laser

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of the Chinese patent application number "202010735260.9" filed by Tsinghua University on July 28, 2020, with the invention titled "A Method and System for Controlling Drug Release Using Pulsed Laser".

technical field

The present application belongs to the technical field of pulsed laser applications, and particularly relates to a method and system for controlling drug release by using pulsed lasers.

Background technique

Cancer is a serious threat to human life and health. Currently, common treatment methods include surgery, radiotherapy, and chemotherapy. Multiple invasive drug injections to cancerous tissue cause great damage to the body, so clinical research on drug delivery systems has received extensive attention. The existing biodegradable drug delivery systems are limited in clinical application due to their poor controllability of drug release time, drug release amount and drug release location. In the prior art, the patent application "A black phosphorus-based hydrogel near-infrared light controllable release immune drug system" (Chinese patent, application number: 201910473466.6) proposes a method for controlling drug release by continuous near-infrared light irradiation, The control of the drug release time is realized. However, in this patent application, due to the large average power of the continuous laser and the poor controllability of the irradiation energy and irradiation position of the continuous laser, the controllability of the drug release amount and the drug release position is poor. Therefore, it is still insufficient in clinical application.

SUMMARY OF THE INVENTION

The purpose of this application is to propose a method and system for controlling drug release by using pulsed laser. In view of the problems existing in the existing methods, using pulsed laser to control the release of drug, because the single-pulse energy of the pulsed laser is adjustable and the number of pulses is controllable, The photothermal reaction of the photosensitive drug release solution can be controlled, and the precise control of drug release amount and release position can be realized.

The method for controlling drug release by using pulsed laser proposed in this application includes the following steps:

(1) Synthesize photosensitive drug release solution, the process is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder is placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution is 0.3-5 mg/ml. The black phosphorus solution is subjected to ultrasonic treatment for 5-8 hours, and the ultrasonic frequency is 3-7 kHz. Perform centrifugal separation at a temperature of 6 to 10° C. for 500 to 1000 seconds, and the centrifugal separation speed is 1500 to 2500 rpm. After standing for 5 to 10 minutes, the supernatant is extracted, and the supernatant is dried to obtain nano black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

Put the nano black phosphorus flakes in water, so that the concentration of the nano black phosphorus solution is 0.01～0.35mg/ml, add the polyethylene glycol amine solution with a concentration of 0.01～0.35mg/ml to the nano black phosphorus solution, the mass of the added The volume ratio is: nanometer black phosphorus solution: polyethylene glycol amine solution=1:(0.8～1.2), obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5～3 hours, the ultrasonic frequency is 3～7kHz, After ultrasonication, magnetic stirring is performed for 1 to 4 hours, and the magnetic stirring speed is 1000 to 1800 rpm;

(1-3) Add an anticancer drug solution with a mass volume concentration of 4 to 8 mg/ml to the first mixed solution in step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: first mixed solution: anticancer drug solution=1:(1～1.5);

(1-4) The second mixed solution of step (1-3) is heated to 63-75° C., agarose is added, and the mass percentage of the agarose added is 0.2-18.0 wt %. After the agarose is completely dissolved, it forms after cooling. Hydrogel to prepare photosensitive drug release solution;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens, so that the number of laser pulses is 30-155, and the single pulse energy of the pulsed laser is 1-4 J/cm ² to control the release of the drug.

The system for controllable drug release based on pulsed laser proposed in this application includes: a pulsed laser, a density attenuator, an electronically controlled shutter, a mirror, an objective lens and an electronically controlled translation stage; the laser pulse sequence emitted by the pulsed laser sequentially passes through the density The attenuation sheet, the electronically controlled shutter, the reflecting mirror and the objective lens are then focused into the photosensitive drug release solution, and the photosensitive drug release solution is placed on the electronically controlled translation stage.

A method and system for controlling drug release by using pulsed laser proposed in this application has the following advantages:

1. The method for controlling drug release by pulsed laser in the present application, because the pulse number of the pulsed laser used is controllable, the drug release can be controlled quantitatively by the number of pulses, and the strict control of the drug release amount is realized, which overcomes the existing problems. The disadvantage of poor controllability of the continuous laser drug delivery system improves the reliability of clinical applications.

2. The application uses pulsed laser to control the drug release system. By adjusting the density attenuation sheet, the single pulse energy incident into the photosensitive drug release solution is strictly controlled to be 1-4J/cm ² . Adjust and irradiate the photosensitive drug release solution to achieve precise control of drug release.

Additional aspects and advantages of the present application will be set forth, in part, in the following description, and in part will be apparent from the following description, or learned by practice of the present application.

Description of drawings

The above and/or additional aspects and advantages of the present application will become apparent and readily understood from the following description of embodiments taken in conjunction with the accompanying drawings, wherein:

FIG. 1 is a schematic structural diagram of a system for controlling drug release by using a pulsed laser proposed in the present application.

FIG. 2 is a schematic diagram of the encapsulation and release process of anticancer drugs.

In Figures 1 and 2, 1 is a pulsed laser, 2 is a density attenuator, 3 is an electronically controlled shutter, 4 is a mirror, 5 is an objective lens, 6 is a photosensitive drug release solution, 7 is an electronically controlled translation, and 8 is agarose water Gel, 9 are anticancer drug particles, 10 are PEGylated black phosphorus nanosheets.

detailed description

The method for controlling drug release by using pulsed laser proposed in this application includes the following steps:

(1) Synthesize photosensitive drug release solution, the process is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder is placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution is 0.3-5 mg/ml. The black phosphorus solution is subjected to ultrasonic treatment for 5-8 hours, and the ultrasonic frequency is 3-7 kHz. Carry out centrifugal separation at a temperature of 6 to 10 ° C for 500 to 1000 seconds, and the centrifugal separation speed is 1500 to 2500 rpm. After standing for 5 to 10 minutes, the supernatant is extracted, and the supernatant is dried to obtain nano black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

Put the nano black phosphorus flakes in water, so that the concentration of the nano black phosphorus solution is 0.01～0.35mg/ml, add the polyethylene glycol amine solution with a concentration of 0.01～0.35mg/ml to the nano black phosphorus solution, the mass of the added The volume ratio is: nanometer black phosphorus solution: polyethylene glycol amine solution=1:(0.8～1.2), obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5～3 hours, the ultrasonic frequency is 3～7kHz, After ultrasonication, magnetic stirring is performed for 1 to 4 hours, and the magnetic stirring speed is 1000 to 1800 rpm;

(1-3) Add an anticancer drug solution with a mass volume concentration of 4 to 8 mg/ml to the first mixed solution in step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: first mixed solution: anticancer drug solution=1:(1～1.5);

(1-4) The second mixed solution of step (1-3) is heated to 63-75° C., agarose is added, and the mass percentage of the agarose added is 0.2-18.0 wt %. After the agarose is completely dissolved, it forms after cooling. Hydrogel to prepare photosensitive drug release solution;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens, so that the number of laser pulses is 30-155, and the single pulse energy of the pulsed laser is 1-4 J/cm ² to control the release of the drug.

The structure of the controllable drug release system based on pulsed laser proposed in this application is shown in FIG. 1 , including: a pulsed laser 1 , a density attenuator 2 , an electronically controlled shutter 3 , a mirror 3 , an objective lens 5 and an electronically controlled translation stage 7 . The laser pulse sequence emitted by the pulsed laser 1 passes through the density attenuating sheet 2, the electronically controlled shutter 3, the mirror 4 and the objective lens 5 in turn and is focused into the photosensitive drug release solution 6, and the photosensitive drug release solution 6 is placed in the described on the electronically controlled translation stage 7.

In the above system, the center wavelength of the pulsed laser is 780-1100 nm, the repetition frequency is 3-9 Hz, and the magnification of the objective lens is 10 times.

The working process of the system based on pulsed laser controllable drug release proposed in this application is as follows:

First, a photosensitive drug release solution is synthesized, and the black phosphorus powder is placed in water to obtain a black phosphorus solution. 3 to 7 kHz, after the end of ultrasonication, centrifuge at a temperature of 6 to 10 °C for 500 to 1000 seconds, and the centrifugal separation speed is 1500 to 2500 rpm. After standing for 5 to 10 minutes, extract the supernatant, and dry the supernatant. dry to obtain nano black phosphorus flakes; put the nano black phosphorus flakes in water, so that the concentration of the nano black phosphorus solution is 0.01～0.35mg/ml, add polyethylene with a concentration of 0.01～0.35mg/ml to the nano black phosphorus solution Diolamine solution, the mass-to-volume ratio added is: nano-black phosphorus solution: polyethylene glycol amine solution=1: (0.8～1.2), to obtain a first mixed solution, and ultrasonically process the first mixed solution for 0.5 to 3 hours , the ultrasonic frequency is 3-7 kHz, the magnetic stirring is 1-4 hours after the ultrasonic, and the magnetic stirring speed is 1000-1800 rpm; in the first mixed solution of the step, the anti-cancer drug solution with a concentration of 4-8 mg/ml is added to obtain the second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: the first mixed solution: the anticancer drug solution=1:(1～1.5), to obtain the second mixed solution; the second mixed solution is heated to 63～ 75 ℃, add agarose, the mass percentage of the added agarose is 0.2-18.0wt%, after the agarose is completely dissolved, a hydrogel is formed after cooling, and the photosensitive drug release solution is prepared; The decay rate and the exposure time of the electronically controlled shutter enable the release of anticancer drugs. Select a suitable objective lens 5, fix the photosensitive drug release solution on the electronically controlled translation stage, adjust the position of the electronically controlled translation stage to focus the laser in the solution; adjust the density attenuation sheet to make the single pulse incident on the photosensitive drug release solution. The energy is controlled at 1-4J/cm ² ; the repetition frequency of the pulsed laser and the exposure time of the electronically controlled shutter are set so that the number of laser pulses passing through the shutter is 30-155, and the photosensitive controlled drug release is realized.

The embodiments of the present application are described below: the black phosphorus powder used in the embodiments of the present application is produced by Beijing Beike New Material Technology Co., Ltd., and the product model is BKTMDC010699.

Example 1:

(1) The process of synthesizing the photosensitive drug release solution is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder was placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution was 0.8 mg/ml. The black phosphorus solution was subjected to ultrasonic treatment for 5 hours, and the ultrasonic frequency was 3.5 kHz. Centrifugal separation was carried out for 550 seconds at a centrifugal separation speed of 1500 rpm, and after standing for 6 minutes for precipitation, the supernatant was extracted, and the supernatant was dried to obtain nano-black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

The nano black phosphorus flakes are placed in water, so that the concentration of the nano black phosphorus solution is 0.05 mg/ml, and the polyethylene glycol amine solution with a concentration of 0.05 mg/ml is added to the nano black phosphorus solution, and the mass volume ratio added is: Nano black phosphorus solution: polyethylene glycol amine solution=1:1, obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5 hours, the ultrasonic frequency is 3.5kHz, and after the ultrasonic is finished, magnetic stirring is performed for 1 hour, and the magnetic stirring speed is is 1200rpm;

(1-3) adding a concentration of 5mg/ml anticancer drug solution to the first mixed solution of step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: the first Mixed solution: in anticancer drug solution=1:1, the third mixed solution is obtained;

(1-4) The third mixed solution of step (1-3) is heated to 65°C, agarose with a sol temperature of 48°C is added, the mass percentage of the added agarose is 5.0 wt %, and when the agarose is completely dissolved, cooled After forming a hydrogel, a photosensitive drug release solution is prepared;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens 5 .

(3) The parameters of the pulsed laser are set as follows: the number of laser pulses is 40, the single-pulse energy of the pulsed laser is controlled to be 2J/cm ² , and the drug release amount is 3.22 mg/ml.

Example 2:

(1) The process of synthesizing the photosensitive drug release solution is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder was placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution was 1 mg/ml. The black phosphorus solution was subjected to ultrasonic treatment for 5.5 hours, and the ultrasonic frequency was 4 kHz. Centrifuge for 600 seconds at a centrifugal speed of 2000 rpm, and after standing for 6 minutes for precipitation, extract the supernatant, and dry the supernatant to obtain nano black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

The nano black phosphorus flakes are placed in water, so that the concentration of the nano black phosphorus solution is 0.08 mg/ml, and the polyethylene glycol amine solution with a concentration of 0.08 mg/ml is added to the nano black phosphorus solution, and the mass volume ratio added is: Nano black phosphorus solution: polyethylene glycol amine solution=1:1.1, obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5 hour, the ultrasonic frequency is 4kHz, after the ultrasonic is finished, magnetic stirring is 1 hour, and the magnetic stirring speed is 1400rpm;

(1-3) adding a concentration of 5mg/ml anticancer drug solution to the first mixed solution of step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: the first Mixed solution: in anticancer drug solution=1:1, the third mixed solution is obtained;

(1-4) The third mixed solution of step (1-3) is heated to 70°C, agarose with a sol temperature of 52°C is added, and the mass percentage of the added agarose is 10.0 wt %. When the agarose is completely dissolved, it is cooled down. After forming a hydrogel, a photosensitive drug release solution is prepared;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens.

(3) The parameters of the pulsed laser are set as follows: the number of laser pulses is 60, the single-pulse energy of the pulsed laser is controlled to be 2.5 J/cm ² , and the drug release amount is 4.17 mg/ml.

Example 3:

(1) The process of synthesizing the photosensitive drug release solution is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder is placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution is 3 mg/ml. The black phosphorus solution is subjected to ultrasonic treatment for 7 hours, and the ultrasonic frequency is 6 kHz. Centrifuge for 800 seconds at a centrifugal speed of 2500 rpm, and after standing for 6 minutes for precipitation, extract the supernatant, and dry the supernatant to obtain nano black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

Put the nano black phosphorus flakes in water so that the concentration of the nano black phosphorus solution is 0.3 mg/ml, and add a polyethylene glycol amine solution with a concentration of 0.3 mg/ml to the nano black phosphorus solution, and the added mass volume ratio is: Nano black phosphorus solution: polyethylene glycol amine solution=1:1.2, obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 1 hour, the ultrasonic frequency is 6 kHz, and after the ultrasonic is finished, magnetic stirring is performed for 1.5 hours, and the magnetic stirring speed is 1800rpm;

(1-3) adding a concentration of 6mg/ml anticancer drug solution to the first mixed solution of step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: the first Mixed solution: in anticancer drug solution=1:1, the third mixed solution is obtained;

(1-4) The third mixed solution of step (1-3) was heated to 68°C, agarose with a sol temperature of 47°C was added, and the mass percentage of the added agarose was 17.0 wt %. When the agarose was completely dissolved, it was cooled down. After forming a hydrogel, a photosensitive drug release solution is prepared;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens.

(3) The parameters of the pulsed laser are set as follows: the number of laser pulses is 100, the single-pulse energy of the pulsed laser is controlled to be 3 J/cm ² , and the drug release amount is 4.86 mg/ml.

Example 4:

(1) The process of synthesizing the photosensitive drug release solution is as follows:

(1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

The black phosphorus powder is placed in water to obtain a black phosphorus solution, the mass volume concentration of the black phosphorus solution is 2mg/ml, the black phosphorus solution is subjected to ultrasonic treatment for 6 hours, and the ultrasonic frequency is 4.2kHz. Carry out centrifugation for 800 seconds, the centrifugation speed is 2000rpm, and after standing for 6 minutes, the supernatant is extracted, and the supernatant is dried to obtain nano black phosphorus flakes;

(1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

Put the nano black phosphorus flakes in water, so that the concentration of the nano black phosphorus solution is 0.2 mg/ml, and add a polyethylene glycol amine solution with a concentration of 0.2 mg/ml to the nano black phosphorus solution, and the added mass volume ratio is: Nano black phosphorus solution: polyethylene glycol amine solution=1:1, obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5 hours, the ultrasonic frequency is 4.2kHz, and the magnetic stirring is performed for 1 hour after the ultrasonication, and the magnetic stirring speed is 1800rpm;

(1-3) adding a concentration of 7mg/ml anticancer drug solution to the first mixed solution of step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: the first Mixed solution: in anticancer drug solution=1:1, the third mixed solution is obtained;

(1-4) The third mixed solution of step (1-3) is heated to 66°C, agarose with a sol temperature of 49°C is added, the mass percentage of the added agarose is 12.0 wt %, and when the agarose is completely dissolved, cooling After forming a hydrogel, a photosensitive drug release solution is prepared;

(2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens.

(3) The parameters of the pulsed laser are set as follows: the number of laser pulses is 140, the single-pulse energy of the pulsed laser is controlled to be 4 J/cm ² , and the drug release amount is 5.05 mg/ml.

The above-mentioned embodiment is the preferred embodiment of the application, but the embodiment of the application is not limited by the above-mentioned embodiment, and any other changes, modifications, substitutions, combinations, Simplification, all should be equivalent substitution methods, and all are included in the protection scope of the present application.

Claims (3)

1. A method for controlling drug release by using pulsed laser, characterized in that, the method comprises the following steps:

   (1) Synthesize photosensitive drug release solution, the process is as follows:

   (1-1) Preparation of black phosphorus nanosheets by solution exfoliation:

   The black phosphorus powder is placed in water to obtain a black phosphorus solution. The mass volume concentration of the black phosphorus solution is 0.3-5 mg/ml. The black phosphorus solution is subjected to ultrasonic treatment for 5-8 hours, and the ultrasonic frequency is 3-7 kHz. Carry out centrifugal separation at a temperature of 6 to 10 ° C for 500 to 1000 seconds, and the centrifugal separation speed is 1500 to 2500 rpm. After standing for 5 to 10 minutes, the supernatant is extracted, and the supernatant is dried to obtain nano black phosphorus flakes;

   (1-2) Coating polyethylene glycol amine on the surface of nano black phosphorus flakes:

   Put the nano black phosphorus flakes in water, so that the concentration of the nano black phosphorus solution is 0.01～0.35mg/ml, add the polyethylene glycol amine solution with a concentration of 0.01～0.35mg/ml to the nano black phosphorus solution, the mass of the added The volume ratio is: nanometer black phosphorus solution: polyethylene glycol amine solution=1:(0.8～1.2), obtain the first mixed solution, carry out ultrasonic treatment to the first mixed solution for 0.5～3 hours, the ultrasonic frequency is 3～7kHz, After ultrasonication, magnetic stirring is performed for 1 to 4 hours, and the magnetic stirring speed is 1000 to 1800 rpm;

   (1-3) Add an anticancer drug solution with a mass volume concentration of 4 to 8 mg/ml to the first mixed solution in step (1-2) to obtain a second solution, the volume ratio of the first mixed solution to the anticancer drug solution is: first mixed solution: anticancer drug solution=1:(1～1.5);

   (1-4) The second mixed solution of step (1-3) is heated to 63-75° C., agarose is added, and the mass percentage of the agarose added is 0.2-18.0 wt %. After the agarose is completely dissolved, it forms after cooling. Hydrogel to prepare photosensitive drug release solution;

   (2) Focus the laser pulse sequence into the photosensitive drug release solution through the objective lens, so that the number of laser pulses is 30-155, and the single pulse energy of the pulsed laser is 1-4 J/cm ² to control the release of the drug.
2. A system for controllable drug release based on pulsed laser, characterized in that the system comprises: a pulsed laser, a density attenuator, an electronically controlled shutter, a mirror, an objective lens and an electronically controlled translation stage; the laser pulse sequence emitted by the pulsed laser After passing through the density attenuation sheet, the electronically controlled shutter, the reflecting mirror and the objective lens in sequence, the photosensitive drug releasing solution is focused into the photosensitive drug releasing solution, and the photosensitive drug releasing solution is placed on the said electronically controlled translation stage.
3. The system for controlled drug release based on pulsed laser according to claim 2, characterized in that the center wavelength of the pulsed laser is 780-1100 nm, the repetition frequency is 3-9 Hz, and the magnification of the objective lens is 10 times.

Images