阅读说明：本技术 微针释药装置及制造方法、皮肤疾病治疗装置 (Microneedle drug delivery device, manufacturing method thereof and skin disease treatment device ) 是由 钱志勇 郝颖 于 2020-04-09 设计创作，主要内容包括：本发明公开了涉及生物医药领域,尤其是微针释药装置制造方法,包括以下步骤：A、通过双乳化法将治疗药物和载药物质制成释药装置；制作微针系统载体材料；B、将释药装置加入微针模板中,然后将微针系统载体材料填充微针模板；C、干燥后取出得到微针释药装置。通过在载药粒子中添加光热转化剂,从而使得释药装置能够吸收特定波长的光,将含有治疗药物的载药粒子输送至疾病组织中,载药粒子释放,当用光照射时,载药粒子中的光热转化剂将光能转化成热能,不仅能使疾病组织高热从而杀伤病灶,而且还可以控制载药粒子中治疗药物的释放行为,达到化疗与光热治疗协同治疗的目的。(The invention discloses a method for manufacturing a microneedle drug release device, which relates to the field of biomedicine and comprises the following steps: A. preparing the therapeutic drug and the drug-loaded substance into a drug release device by a double emulsification method; manufacturing a microneedle system carrier material; B. adding the drug release device into the microneedle template, and then filling the microneedle template with the microneedle system carrier material; C. taking out the microneedle drug delivery device after drying. Through adding light-heat conversion agent in carrying medicine particle to make medicine release device can absorb the light of specific wavelength, carry the medicine carrying particle who contains treatment medicine to the disease tissue in, carry the medicine carrying particle release, when shining with the light, light-heat conversion agent in the medicine carrying particle converts light energy into heat energy, thereby can not only make the high fever of disease tissue kill and kill the focus, but also can control the release action of treatment medicine in the medicine carrying particle, reach the purpose of chemotherapy and light-heat treatment concurrent therapy.)

1. The microneedle drug release device is characterized in that: the drug delivery device comprises a treatment drug and drug-loaded particles, wherein the treatment drug is loaded in the drug-loaded particles, and the drug-loaded particles contain a photothermal conversion agent.

2. A microneedle delivery device as claimed in claim 1, wherein: the photothermal conversion agent has near-infrared light responsiveness.

3. A microneedle delivery device as claimed in claim 2, wherein: the drug-loaded particles can release therapeutic drugs under the irradiation of near-infrared light of 780-820 nm; the photothermal conversion agent comprises one or more of nanogold rods, indocyanine green, neoindocyanine green and IR780 iodide.

4. A microneedle delivery device as claimed in claim 1, wherein: the therapeutic agent is used for treating skin diseases; the skin diseases include infection, cancer; the cancer comprises skin squamous carcinoma, skin adenocarcinoma, melanoma; the therapeutic drug is one or more of 5-fluorouracil, dacarbazine, doxorubicin, docetaxel and paclitaxel.

5. A microneedle delivery device as claimed in claim 1, wherein: the drug-loaded particles are nano-scale particles; the drug-carrying particles are made of two-block polymer methoxy polyethylene glycol-polycaprolactone, methoxy polyethylene glycol-polylactic acid or a composition thereof; in the two-block polymer methoxy polyethylene glycol-polycaprolactone or methoxy polyethylene glycol-polylactic acid, the molecular weight of methoxy polyethylene glycol is 4000-6000, preferably 5000; the molecular weight of the polycaprolactone or the polylactic acid is 20000-50000, and preferably 30000.

6. A microneedle delivery device as claimed in claim 5, wherein: the drug-loaded particles are composed of methoxy polyethylene glycol-polycaprolactone or methoxy polyethylene glycol-polylactic acid, and the block ratio of the drug-loaded particles is 1: 4-1: 10.

7. A microneedle delivery device as claimed in claim 1, wherein: the microneedle system is a microneedle array consisting of a plurality of microneedles; the height of a single microneedle is 400-800 μm, preferably 600 μm; the molecular weight of the microneedle material is 400-2000 kDa.

8. A microneedle delivery device as claimed in claim 1, wherein: the micro-needle system is prepared by taking sodium hyaluronate, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, sodium alginate or a composition thereof as a carrier material.

9. The manufacturing method of the microneedle drug release device is characterized in that: the method comprises the following steps:

A. preparing the therapeutic drug and the drug-loaded substance into a drug release device by a double emulsification method;

B. adding the drug release device into the microneedle template, and then filling the microneedle template with the microneedle system carrier material;

C. drying and then taking out to obtain the microneedle drug delivery device as claimed in any of claims 1 to 8.

10. The method for manufacturing a microneedle delivery device according to claim 9, wherein: in step a, the emulsifier is polyvinyl alcohol.

11. The method for manufacturing a microneedle delivery device according to claim 9, wherein: the ratio of the drug release device to the carrier material of the microneedle system is 1: 1-1: 20.

12. A dermatological disease treatment device, characterized in that: comprises the microneedle drug delivery device as claimed in any of claims 1 to 8 and a near infrared illumination device.

Technical Field

The invention relates to the field of biological medicine, in particular to a microneedle drug release device, a manufacturing method and a skin disease treatment device.

Background

The epidermic diseases comprise microbial infection, canceration, injury and the like, and in the case of epidermic cancer, the epidermic cancer is a malignant tumor of the skin, and the incidence rate of the epidermic cancer is gradually increased along with the aging of the population in China. According to the source of tumor cells, malignant tumors of epidermis can be classified into squamous cell carcinoma, basal cell carcinoma, malignant melanoma, etc. Superficial malignant tumor of epidermis is usually treated by surgical resection, radiotherapy, chemotherapy, laser therapy, etc. The surgical therapy may not completely remove the residual tumor cells, and thus the tumor is easy to recur. Radiotherapy and chemotherapy have certain treatment effect, but serious toxic and side effects are generated in the treatment process, and the immune system of the organism is damaged. Laser therapy is an emerging treatment, but long-term laser therapy causes DNA damage to the skin and, again, leads to the development of epidermal carcinoma. In summary, these conventional treatments all have significant therapeutic limitations.

The micro-needle (Microneedles) is a micro-needle point structure manufactured by micro-nano processing technology, and has the size of micron, the height of 10-2000 μm and the width of 10-50 μm. Currently, microneedle drug delivery systems have been widely used for delivery of biological macromolecules such as drugs, proteins, genes, RNA, vaccines, etc. due to their rapid, efficient, and painless modes of administration. Compared with an oral administration system and an injection administration system, the microneedle administration system can directly penetrate through the stratum corneum of the skin to realize the transmission of the drug and avoid the first pass effect of the liver. Combining the characteristics of epidermal tumor, the micro-needle drug delivery system is a better drug delivery carrier applied to the treatment of epidermal cancer. Although microneedle administration has many advantages, the rate of drug release is difficult to control, and the temperature at which the microneedles penetrate into the lesion cannot be controlled.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a drug release device which can control the drug release behavior and change the temperature of the focus, and can be used for treating various skin diseases by the cooperation of photothermal treatment and chemotherapy.

The microneedle drug release device used for solving the technical problem comprises a dissolvable microneedle system with skin penetrating capability, wherein the microneedle system is internally provided with the drug release device, the drug release device comprises a treatment drug and drug-loaded particles, the treatment drug is loaded in the drug-loaded particles, and the drug-loaded particles contain a photo-thermal conversion agent.

The micro-needle system contains a large number of micro-needle point structures, the micro-needle can be dissolved in a human body, and the micro-needle can directly penetrate through the cuticle or other tissues of the skin to realize the transmission of a medicine release device, so that the medicine can directly reach the focus, and the first pass effect of the liver is avoided. The therapeutic drug is mainly selected according to the types of diseases, the drug-loaded particles are used as a carrier of the therapeutic drug and are used as a tiny drug delivery unit after the therapeutic drug is loaded, the size of the drug-loaded particles can be determined according to the actual diseases, and the drug-loaded particles are usually nano-sized, so that the drug-loaded particles can effectively act on the focus. The basic material of the drug-carrying particles is a high molecular material with good biocompatibility and biodegradability, so that the drug-carrying particles can be degraded after entering a human body, thereby releasing the treatment drugs therein.

Further, the photothermal conversion agent has near-infrared light responsiveness. The near infrared light has the characteristics of high tissue permeability and minimum phototoxicity, and the photothermal conversion agent with near infrared light responsiveness is adopted, so that the drug release device can control the release behavior of the therapeutic drug in the skin, and simultaneously can generate heat at the drug release device, and the focus is killed and killed by high heat.

Specifically, the drug-loaded particles can release therapeutic drugs under the irradiation of near-infrared light of 780-820 nm; the photothermal conversion agent comprises one or more of nanogold rods, indocyanine green, neoindocyanine green (IR820) and IR780 iodide.

In particular, the therapeutic agent is used for treating skin diseases; the skin diseases include infection, cancer; the cancer comprises skin squamous carcinoma, skin adenocarcinoma, melanoma; the therapeutic drug is one or more of 5-fluorouracil, dacarbazine, doxorubicin, docetaxel and paclitaxel.

Specifically, the drug-loaded particles are nanoscale particles; the drug-carrying particles are made of methoxy polyethylene glycol-polycaprolactone (MPEG-PCL), methoxy polyethylene glycol-polylactic acid (MPEG-PDLLA) or a composition thereof, and in the two-block polymer methoxy polyethylene glycol-polycaprolactone or methoxy polyethylene glycol-polylactic acid, the molecular weight of methoxy polyethylene glycol is 4000-6000, preferably 5000; the molecular weight of the polycaprolactone or the polylactic acid is 20000-50000, and preferably 30000. The molecular weights of the two blocks in the two-block polymer are 5000-20000, 5000-30000, 5000-40000 and 5000-50000 respectively; for example, in methoxy polyethylene glycol-polycaprolactone, the methoxy polyethylene glycol is 5000, and the polycaprolactone is 20000-50000; for example, in the methoxypolyethylene glycol-polylactic acid, the methoxypolyethylene glycol is 5000, and the polylactic acid is 20000 to 50000.

Specifically, the drug-loaded particles are composed of methoxy polyethylene glycol-polycaprolactone or methoxy polyethylene glycol-polylactic acid, and the block ratio of the drug-loaded particles is 1: 4-1: 10.

Specifically, the height of the microneedles in the microneedle system may be 400-800 μm, preferably 600 μm; the molecular weight of the microneedle material is 400-2000 kDa.

Specifically, the microneedle system is prepared from sodium hyaluronate, chitosan, polyvinylpyrrolidone, polyvinyl alcohol, sodium alginate or a composition thereof.

The invention also aims to solve the technical problem of providing a manufacturing method for manufacturing the microneedle drug release device with photoresponse, which comprises the following steps: the method comprises the following steps:

A. the treatment drug and the drug-carrying substance are prepared into the drug release device by a double emulsification method; preparing a microneedle system carrier material;

B. adding the drug release device into the microneedle template, and then filling the microneedle template with the microneedle system carrier material;

C. and taking out the microneedle drug release device after drying.

Specifically, in step a, the emulsifier is polyvinyl alcohol.

Specifically, the ratio of the drug release device to the carrier material of the microneedle system can be 1: 1-1: 20.

Another technical problem to be solved by the present invention is to provide a skin disease treatment device capable of controlling drug release behavior and changing the temperature of the focus, comprising the microneedle drug release device and the near infrared illumination device. When the skin disease treatment device is adopted, the microneedle of the microneedle drug release device is penetrated into the diseased tissue, the drug release device is released along with the dissolution of the microneedle, the near infrared light emitted by near infrared illumination equipment is utilized to irradiate the diseased tissue, and the photothermal conversion agent in the drug-carrying particles converts the light energy into heat energy, so that the diseased tissue is heated to kill tumor cells, the release behavior of the treatment drug in the drug-carrying particles can be controlled, and the purpose of chemotherapy and photothermal treatment cooperative treatment is achieved.

The near-infrared illumination equipment can be a common equipment in the field, namely a multimode fiber coupled laser, and can output near-infrared light of 780-820 nm. During treatment, the output laser of the near-infrared illumination equipment corresponds to the maximum ultraviolet absorption peak of the photothermal conversion agent in the drug-loaded particles. When the maximum absorption wavelength of the photothermal conversion agent in the used drug-loaded particles is 808nm, the following parameters are recommended in the treatment: the using amount of the photo-thermal conversion agent is 0-1 mg; the 808nm near infrared light irradiation power is 0-5W/cm²(ii) a The temperature change of the 808nm near-infrared light after irradiating the microneedle is 30-80 ℃; the 808nm near infrared light irradiation time is 1-60 min.

The invention has the beneficial effects that:

through adding light-heat conversion agent in the medicine carrying particle for the medicine release device can absorb the light of specific wavelength, will contain the medicine carrying particle of treatment medicine and carry to the disease tissue in, the medicine carrying particle release, when shining with light, light-heat conversion agent in the medicine carrying particle converts light energy into heat energy, thereby can not only make the high fever of disease tissue kill and kill the focus, but also can control the release action of treatment medicine in the medicine carrying particle, reaches the purpose of chemotherapy and light-heat treatment concurrent therapy. The mode is particularly convenient and effective in treating skin diseases, the invention also utilizes the micro-needle to convey the drug-loaded particles into the disease tissues, and simultaneously utilizes near infrared light with high tissue permeability and minimum phototoxicity as a light source. Compared with the traditional treatment methods of skin squamous carcinoma, skin adenocarcinoma and melanoma, the near-infrared light-responsive drug delivery device based on the microneedle, which is constructed by the invention, can directly penetrate through the horny layer of the skin and enter the dermis through the acanthocyte layer and the basal cell layer so as to realize the transmission of the drug and avoid the first pass effect of the liver. Compared with the methods of surgical excision, radiotherapy, chemotherapy, laser therapy and the like, the near-infrared light-responsive drug release device has the advantages of small toxic and side effects, high compliance, convenient operation and the like.

Drawings

Figure 1 is a schematic view of the method of preparation and the drug release behaviour of a drug release device according to the invention;

FIG. 2 is a particle size distribution plot in drug-loaded particle characterization;

FIG. 3 is a transmission electron micrograph of a drug loaded particle characterization;

FIG. 4 is a graph of UV absorption in drug-loaded particle characterization;

FIG. 5 is a graph of release behavior in drug-loaded particle characterization;

fig. 6 is a flow chart of a manufacturing method of a microneedle drug release device;

fig. 7 is a representation of microneedles;

fig. 8 is a trypan blue staining graph and H & E section graph of the skin penetration ability of the microneedles;

FIG. 9 is a microneedle photothermal effect profile;

fig. 10 is a graph of temperature rise of microneedles;

FIG. 11 is a graph of tumor growth for the examples;

FIG. 12 is a graph of the change in body weight of experimental mice;

FIG. 13 is a graph of tumor size comparison;

FIG. 14 is a graph of tumor weight;

illustration of the drawings: MPEG-PCL is a drug-carrying particle of a methoxy polyethylene glycol-polycaprolactone two-block polymer; the 5-Fu-MPEG-PCL is a drug-loaded nanoparticle only containing 5-fluorouracil; ICG-MPEG-PCL is a nano drug-carrying particle only containing indocyanine green; the 5-Fu & ICG-MPEG-PCL is a nano drug-loaded particle loaded with 5-fluorouracil and indocyanine green; control is a Control without any treatment; HA MN is a pure hyaluronic acid dissolvable microneedle group; the ICG @ HA MN is a hyaluronic acid dissoluble microneedle group loaded with ICG; 5-Fu-MPEG-PCL @ HA MN is a hyaluronic acid dissoluble microneedle group loaded with 5-Fu-MPEG-PCL; ICG-MPEG-PCL @ HA MN is a hyaluronic acid dissoluble microneedle group loaded with ICG-MPEG-PCL; the 5-Fu & ICG-MPEG-PCL @ HA MN is a hyaluronic acid dissoluble microneedle group loaded with the 5-Fu & ICG-MPEG-PCL; fig. 11 to 14 are a set of diagrams in which the correspondence of the labels is on fig. 14.

Detailed Description

The invention will be further explained with reference to the drawings.

The nomenclature abbreviated, methoxypolyethylene glycol is MPEG, polycaprolactone is PCL, polylactic acid PDLLA, 5-fluorouracil is 5-Fu, and indocyanine green is ICG.

Referring to fig. 6, the method for manufacturing the microneedle device is described in the following examples, and it is understood that suitable process parameters may be selected according to different factors such as diseases to be treated, treatment sites, and raw materials.

Firstly, preparing a therapeutic drug, a drug-loaded substance and a micro-needle system carrier material, wherein the therapeutic drug and the drug-loaded substance are prepared into a drug release device by a double-emulsion method. The drug-carrying substance for forming the drug-carrying particle can be purchased directly or prepared by self, and if the drug-carrying substance is prepared by self, the drug-carrying substance can be prepared by the following method.

The preparation method of the drug-loaded substance comprises the following steps: weighing appropriate amount of polyethylene glycol monomethyl ether and caprolactone or D, L-lactide, adding into a three-necked bottle, vacuumizing the reaction system for 3min, introducing nitrogen for 3min, and repeating for 3 times. Under the protection of nitrogen, adding stannous octoate (0.3% w/w), heating to 150 ℃ under the condition of magnetic stirring, and reacting for 9 hours. After the reaction is finished, the temperature is cooled to room temperature, and a proper amount of dichloromethane (CH) is added₂Cl₂) The crude product was dissolved and then poured into 5 volumes of frozen anhydrous ether and stirred rapidly to produce a precipitate. After the crude product had precipitated completely, the supernatant was removed and the washing was repeated three times. And then, drying the precipitate in a vacuum drying oven at 37 ℃ to constant weight to obtain two-block polymer of methoxy polyethylene glycol-polycaprolactone (MPEG-PCL) and methoxy polyethylene glycol-polylactic acid (MPEG-PDLLA). The molecular weight of the methoxypolyethylene glycol is 4000-6000, preferably 5000; the molecular weight of polycaprolactone or polylactic acid is 20000-50000, and the preferred molecular weight is30000. The molecular weights of the two blocks in the two-block polymer are 5000-20000, 5000-30000, 5000-40000 and 5000-50000 respectively; for example, in methoxy polyethylene glycol-polycaprolactone, the methoxy polyethylene glycol is 5000, and the polycaprolactone is 20000-50000; for example, in the methoxypolyethylene glycol-polylactic acid, the methoxypolyethylene glycol is 5000, and the polylactic acid is 20000 to 50000. The drug-carrying particles are generally made of diblock polymers.

There are four cases of making therapeutic drugs and drug-loaded substances into drug delivery devices by the double emulsion method, each as described separately below:

1. dispersing the water-soluble chemotherapeutic drug or the photo-thermal conversion agent into a water phase, dispersing the material into an organic phase, adding the water phase into the organic phase, and performing ultrasonic emulsification for 5min to obtain a W/O solution; then adding the W/O solution into a polyvinyl alcohol aqueous solution, and carrying out ultrasonic emulsification for 5min to obtain a W/O/W solution; finally, removing the organic solvent in the solution to obtain the nano particles loaded with the chemotherapeutic drug and the photo-thermal conversion agent.

2. Dispersing the fat-soluble chemotherapeutic drug or the photothermal conversion agent and the material into an organic phase, adding the organic phase into a polyvinyl alcohol aqueous solution, and performing ultrasonic emulsification for 5min to obtain an O/W solution; finally, removing the organic solvent in the solution to obtain the nano particles loaded with the chemotherapeutic drug and the photo-thermal conversion agent.

3. Dispersing water-soluble chemotherapeutic drugs into a water phase, dispersing a fat-soluble photothermal conversion agent material into an organic phase, adding the water phase into the organic phase, and performing ultrasonic emulsification for 5min to obtain a W/O solution; then adding the W/O solution into a polyvinyl alcohol aqueous solution, and carrying out ultrasonic emulsification for 5min to obtain a W/O/W solution; finally, removing the organic solvent in the solution to obtain the nano particles loaded with the chemotherapeutic drug and the photo-thermal conversion agent.

4. Dispersing the water-soluble photo-thermal conversion agent into a water phase, dispersing the fat-soluble chemotherapeutic drug into an organic phase, adding the water phase into the organic phase, and performing ultrasonic emulsification for 5min to obtain a W/O solution; then adding the W/O solution into a polyvinyl alcohol aqueous solution, and carrying out ultrasonic emulsification for 5min to obtain a W/O/W solution; finally, removing the organic solvent in the solution to obtain the nano particles loaded with the chemotherapeutic drug and the photo-thermal conversion agent.

Secondly, preparing the dissolvable microneedle system loaded with the near-infrared light-responsive nanoparticles by a template method. The microneedle templates were made of Polydimethylsiloxane (PDMS) and were 1cm × 1cm in size, and each template consisted of 400(20 × 20) tip grooves. The preparation process comprises the following steps: firstly, adding drug-loaded nano particles with a certain concentration into a PDMS micro-needle template, centrifuging at the rotating speed of 3000rpm/min for 30min, and removing redundant nano particles; adding a certain amount of microneedle materials uniformly dissolved in deionized water into a microneedle template, centrifuging at the rotating speed of 3000rpm/min for 30min, and standing overnight in a precision air-blast drying oven at 45 ℃; and finally, separating the obtained dry product from the PDMS microneedle template to obtain the dissolvable microneedle drug release device loaded with the near-infrared light responsive nanoparticles.

The prepared microneedle drug delivery device based on the microneedle has near-infrared light-responsive release behavior, the response wavelengths of the nanogold rods and the indocyanine green are 808nm, the response wavelengths of the indocyanine green (IR820) are 820nm and 780nm respectively, the response wavelength is the wavelength with the optimal absorbance, and the wavelengths near the response wavelength still have better absorbance, which can be shown in figure 4. When irradiated with near infrared light of a corresponding wavelength, for example, near infrared light of 808nm, the photothermal conversion agent in the nanoparticles converts light energy into heat energy, so that the nanoparticles are ruptured, and the chemotherapeutic drug loaded in the nanoparticles is released.

The invention can be used for treating skin squamous carcinoma, skin adenocarcinoma and melanoma by utilizing the microneedle medicament release device. The micro-needle of the micro-needle drug release device is penetrated into the tumor part, the drug-loaded nano-particles are released along with the dissolution of the micro-needle, when the micro-needle is irradiated by near infrared light with the wavelength of 808nm, the photo-thermal conversion agent in the nano-particles converts the light energy into heat energy, thereby not only leading the tumor tissue to have high heat to kill the tumor cells, but also controlling the release behavior of the drugs in the nano-particles and achieving the purpose of the chemotherapy and photo-thermal therapy cooperative treatment.