CN115040470A

CN115040470A - Microneedle patch and method for producing same

Info

Publication number: CN115040470A
Application number: CN202210774707.2A
Authority: CN
Inventors: 杨文龙; 邓静怡; 万昱; 庄嘉琅; 乔正华; 冼依雯
Original assignee: Shenzhen Rongwei Technology Co ltd
Current assignee: Shenzhen Rongwei Technology Co ltd
Priority date: 2022-07-01
Filing date: 2022-07-01
Publication date: 2022-09-13

Abstract

The embodiment of the invention discloses a microneedle patch and a preparation method thereof. The method for manufacturing the microneedle patch of the embodiment of the present invention includes, for example: dissolving a high molecular material in water to obtain a high molecular polymer aqueous solution; adding doxorubicin and copper sulfide nanoparticles into the high-molecular polymer aqueous solution to obtain a microneedle matrix solution; placing the microneedle matrix solution into a microneedle female die, centrifuging and curing to obtain a microneedle matrix layer; and attaching the substrate layer to the microneedle matrix layer to obtain the microneedle patch. The microneedle patch provided by the embodiment of the invention can be used for better treating breast cancer.

Description

Microneedle patch and method for producing same

Technical Field

The invention relates to the technical field of medicines, in particular to a microneedle patch and a preparation method of the microneedle patch.

Background

Worldwide, breast cancer still ranks as the first of female cancers, 167.1 million new cases of breast cancer occur every year worldwide, and about 52.2 million patients die of breast cancer every year. In china, breast cancer is the highest prevalence of cancer in women. Doxorubicin is a first-line therapeutic drug for treating breast cancer, but doxorubicin has a strong systemic effect when administered by injection, and has the disadvantages of myelosuppression, cardiotoxicity, nausea and vomiting. Therefore, there is a need for a better therapeutic drug or method to treat breast cancer.

Disclosure of Invention

Aiming at least partial defects and shortcomings in the prior art, the embodiment of the invention provides a microneedle patch and a preparation method thereof, which can better treat breast cancer.

In one aspect, embodiments of the present invention provide a method for preparing a microneedle patch, for example, including: dissolving a high molecular material in water to obtain a high molecular polymer aqueous solution; adding doxorubicin and copper sulfide nanoparticles into the high-molecular polymer aqueous solution to obtain a microneedle matrix solution; placing the microneedle matrix solution into a microneedle female die, centrifuging and curing to obtain a microneedle matrix layer; and attaching the substrate layer to the microneedle matrix layer to obtain the microneedle patch.

In one embodiment of the present invention, the polymer material includes a first polymer material and a second polymer material; the first high polymer material comprises one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid and chitosan, and the second high polymer material comprises one of monomer polymer or copolymer of carbopol, vinyl pyrrolidone and derivatives thereof.

In an embodiment of the present invention, the mass ratio of the first polymer material to the second polymer material is: (1-3) and (3-10).

In one embodiment of the present invention, the mass ratio of the doxorubicin, the copper sulfide nanoparticles and the polymer material is: (0.05-0.1), (0.3-0.5) and (1).

In one embodiment of the present invention, the preparation method further comprises a copper sulfide nanoparticle preparation step, and the copper sulfide nanoparticle preparation step specifically comprises: adding sodium citrate and copper chloride into deionized water and stirring to obtain an initial solution; adding a sodium sulfide aqueous solution into the initial solution and carrying out high-temperature oscillation to obtain a transition solution; and adding a sodium chloride ethanol aqueous solution into the transition solution at a low temperature, centrifuging, removing supernatant, and washing to obtain the copper sulfide nano-particles.

In one embodiment of the invention, the mass ratio of the sodium citrate to the copper chloride is 1 (1.0-1.2); the mass ratio of sodium sulfide in the sodium sulfide water solution is 20-26%.

In an embodiment of the present invention, the preparation method further includes a microneedle negative mold preparation step, and the microneedle negative mold preparation step specifically includes: in a vacuum environment, the mass ratio is (0.5-3): (5-20) mixing the polydimethylsiloxane curing agent with the prepolymer to obtain a polydimethylsiloxane mixed solution; and casting the polydimethylsiloxane mixed solution on a male mold of a metal microneedle body and solidifying to obtain the microneedle female mold.

In another aspect, embodiments of the present invention provide a microneedle patch, for example, including: a base layer; and a microneedle matrix layer disposed on the base layer; wherein, the microneedle matrix layer comprises a high molecular material, doxorubicin and copper sulfide nanoparticles.

In one embodiment of the present invention, the mass ratio of the doxorubicin, the copper sulfide nanoparticles and the high molecular material is: (0.05-0.1), (0.3-0.5) and (1).

In one embodiment of the present invention, the polymer material includes a first polymer material and a second polymer material; the mass ratio of the first high polymer material to the second high polymer material is as follows: (1-3) and (3-10); the first high polymer material comprises one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid and chitosan, and the second high polymer material comprises one of monomer polymer or copolymer of carbopol, vinyl pyrrolidone and derivatives thereof.

The technical scheme has the following advantages or beneficial effects: according to the embodiment of the invention, doxorubicin and copper sulfide are combined to be used as a preparation, and a transdermal drug delivery mode is adopted to treat tumors, so that the treatment effect of breast cancer is improved, and the toxic and side effects are reduced. The preparation method provided by the embodiment of the invention is simple in process and convenient to prepare. In addition, the microneedle patch prepared by the preparation method provided by the embodiment of the invention has strong mechanical strength and strong skin penetrating power.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without creative efforts.

Fig. 1 is a schematic flow chart of a method for manufacturing a microneedle patch according to an embodiment of the present invention.

Fig. 2 is a schematic flow chart of another method for preparing a microneedle patch according to an embodiment of the present invention.

Fig. 3 is a detailed schematic flow diagram of the preparation of copper sulfide nanoparticles of fig. 2.

Fig. 4 is a detailed flow diagram of the process of fig. 2 for preparing a negative microneedle mold.

Fig. 5 is a schematic structural diagram of a microneedle patch manufactured by the manufacturing method provided by the embodiment of the present invention.

Fig. 6 is a schematic view showing changes in tumor volume of nude mice treated with the microneedles provided in example 1 according to the present invention.

Fig. 7 is a graph showing the effect of pressing the skin surface of the rat skin for 5min after vertically placing a 200g weight on the microneedle in test example 2.

FIG. 8 is a graph showing the change in body weight of four groups of nude mice in experimental example 3.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.

It is to be understood that the terms first, second, end, and the like in the description and in the claims are used for distinguishing between similar elements and not necessarily for describing a particular sequential or chronological order. It is to be understood that the terms so used are interchangeable under appropriate circumstances such that the embodiments of the invention described herein are, for example, capable of operation in sequences other than those illustrated or otherwise described herein. Furthermore, the terms "comprises," "comprising," and "having," and any variations thereof, are intended to cover a non-exclusive inclusion, such that a process, system, article, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, article, or apparatus.

As shown in fig. 1, an embodiment of the present invention provides a method for preparing a microneedle patch. The method for preparing the microneedle patch includes, for example, the steps of:

s100: dissolving a high molecular material in water to obtain a high molecular polymer aqueous solution;

s200: adding doxorubicin and copper sulfide nanoparticles into the high-molecular polymer aqueous solution to obtain a microneedle matrix solution;

s300: placing the microneedle matrix solution into a microneedle female die, centrifuging and curing to obtain a microneedle matrix layer; and

s400: and attaching the substrate layer to the microneedle matrix layer to obtain the microneedle patch.

According to the embodiment of the invention, doxorubicin and copper sulfide are combined to be used as a preparation, and a transdermal drug delivery mode is adopted to treat tumors, so that the treatment effect of breast cancer is improved, and the toxic and side effects are reduced. The copper sulfide nano-particles as a novel nano-preparation have the characteristics of no toxicity, uniform dispersion and high photothermal effect, have better tumor treatment effect, and have a specific mechanism of locally heating tumor parts under the action of near-infrared laser so as to kill tumor cells. The preparation method provided by the embodiment of the invention is simple in process and convenient to prepare.

The polymer material includes, for example, a first polymer material and a second polymer material. The mass ratio of the first high polymer material to the second high polymer material is as follows: (1-3) and (3-10). The first polymer material includes, for example, one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid, and chitosan. The second polymer material includes, for example, one of monomer polymers or copolymers of carbopol, vinyl pyrrolidone and derivatives thereof.

Further, as shown in fig. 2, the method for preparing the microneedle patch of the present embodiment may further include:

s40, preparing the copper sulfide nano particles.

Specifically, as shown in fig. 3, step S40 specifically includes:

s401, adding sodium citrate and copper chloride into deionized water and stirring to obtain an initial solution;

s403, adding a sodium sulfide aqueous solution into the initial solution, and performing high-temperature oscillation to obtain a transition solution; and

and S405, adding a sodium chloride ethanol aqueous solution into the transition solution at a low temperature, performing centrifugal treatment, removing supernatant, and washing to obtain the copper sulfide nano-particles.

Wherein the mass ratio of the sodium citrate to the copper chloride is 1 (1.0-1.2). The mass ratio of sodium sulfide in the sodium sulfide aqueous solution is 20-26%.

s80, preparing a microneedle female die.

Specifically, as shown in fig. 4, step S80 specifically includes:

s801, in a vacuum environment, mixing the components in a mass ratio of (0.5-3): (5-20) mixing the polydimethylsiloxane curing agent with the prepolymer to obtain a polydimethylsiloxane mixed solution;

and S803, casting the polydimethylsiloxane mixed solution on a male mold of the metal microneedle body and curing to obtain the female mold of the microneedle.

The prepolymer mixture is, for example, SYLGARD184 silicone rubber.

The microneedle female die provided by the embodiment of the invention has high die transparency, and if the solution for preparing the microneedle patch is colored, the solution can be conveniently observed whether permeates into the micropores of the die or not after the solution is added into the micropores of the microneedle female die. In addition, the microneedle female die can be repeatedly used, and the cost is reduced. After the microneedle patch is peeled off from the microneedle negative mould, the microneedle negative mould needs to be soaked in purified water for a period of time before being used. The microneedle female die has strong flexibility and can be used for preparing microneedle patches with different heights according to requirements.

In addition, refer to fig. 5, which is a schematic structural diagram of a microneedle patch manufactured by the manufacturing method according to the embodiment of the present invention. Specifically, as shown in fig. 5, the microneedle patch 100 includes, for example, a base layer 120 and a microneedle matrix layer 110. The microneedle matrix layer 110 is disposed on the substrate layer 120.

The base layer 120 is, for example, a single-sided foam tape. The single-sided foam adhesive tape is prepared from ethylene-vinyl acetate copolymer, has a certain heat preservation effect, and prevents heat generated by photothermal therapy from overflowing.

The microneedle matrix layer 110 includes materials such as a polymer material, doxorubicin, and copper sulfide nanoparticles 111. The mass ratio of the doxorubicin, the copper sulfide nano-particles and the high polymer material is as follows: (0.05-0.1), (0.3-0.5) and (1). The polymer material includes, for example, a first polymer material and a second polymer material. The mass ratio of the first high polymer material to the second high polymer material is as follows: (1-3) and (3-10); the first polymer material includes, for example, one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid, and chitosan. The second polymer material includes, for example, one of monomer polymers or copolymers of carbopol, vinyl pyrrolidone and derivatives thereof.

As shown in fig. 5, the microneedle substrate layer 110 of the microneedle patch has a shape such as a cone, a triangular pyramid, or other shapes.

In order to better understand the manufacturing method provided by the embodiments of the present invention, the process of manufacturing the microneedle patch will be described in detail below.

First, a step of preparing copper sulfide nanoparticles. Specifically, the steps include:

(1) in order to improve the forming of copper sulfide nano particles, adding sodium citrate and copper chloride with the mass ratio of 1 (1.0-1.2) into deionized water, and stirring for 0.5-1.0 hour to obtain a light blue clear solution, namely an initial solution;

(2) adding a sodium sulfide aqueous solution with the mass ratio of sodium sulfide (20-26%) into the light blue clear solution, and then oscillating at high temperature for 0.15-0.5 hours to obtain a dark green solution, namely a transition solution, wherein the temperature of the high-temperature oscillation is 80-100 ℃;

(3) transferring the dark green solution to a low-temperature environment with the temperature of 0-4 ℃, adding a sodium chloride ethanol aqueous solution, centrifuging at a high speed, removing a supernatant, and washing with ethanol water to obtain copper sulfide nanoparticles; typically, the ethanol water is in a mixed configuration at an equal volume ratio, such as 1: 1. The mass ratio of sodium chloride to ethanol water in the sodium chloride ethanol water solution is (70-95%): 1.

and secondly, preparing a microneedle female die. Specifically, the steps include:

(1) the preparation method comprises the following steps of (1) mixing an uncrosslinked and cured polydimethylsiloxane curing agent and a prepolymer according to a certain mass ratio (0.5-3): (5-20) mixing, wherein the prepolymer is SYLGARD184 silicone rubber;

(2) removing redundant gas under vacuum to obtain a polydimethylsiloxane mixed solution, pouring the solution on a male mold of a metal microneedle body, and curing at high temperature to obtain a female mold of the microneedle;

and finally, a step of preparing the microneedle patch. Specifically, the steps include:

(1) the mass ratio of (1-3): (3-10) adding the first high polymer material and the second high polymer material into water to obtain a high polymer material aqueous solution, adding doxorubicin and copper sulfide nanoparticles into the high polymer material aqueous solution, performing ultrasonic treatment under a high-temperature environment until the solution is basically clear and uniform, and removing bubbles in the solution by means of high-speed centrifugation and the like to obtain a microneedle matrix solution; wherein the temperature of the high-temperature environment is (80-100 ℃); the mass ratio of the doxorubicin, the copper sulfide nano particles and the high molecular polymer is as follows: (0.05-0.1): 0.3-0.5): 1; in addition, the quality of the high molecular polymer needs to ensure that the chemotherapeutic drug and the nanoparticles are uniformly dispersed in the microneedle matrix solution;

(2) adding the microneedle matrix solution into a microneedle female die, centrifuging, scraping the residual microneedle matrix solution on the microneedle female die, and curing at the temperature of 40-70 ℃ for 2-15 hours to obtain a microneedle matrix layer;

(3) and (3) attaching the single-sided foam adhesive tape to the microneedle substrate layer of the microneedle female die to obtain the microneedle patch.

In addition, the present invention will also illustrate the performance of the microneedle patch provided by the embodiment of the present invention in 3 specific test examples.

Test example 1 microneedle patch having chemotherapy-photothermal combination for treating transplanted mammary tumor in nude mice

20 nude mice for 2 weeks were randomly and evenly distributed into 4 mouse cages to be inoculated with breast cancer transplantation tumor cells, and then raised for 2 weeks to record the size of their tumor volume, and the administration was performed every three days and the volume of their tumor was recorded. Randomly grabbing three nude mice from each group of nude mice for posterior cervical vertebra sacrifice, dissecting the sacrificed nude mice, and taking out the nude mice breast cancer transplantation tumor. The duration period is 24 days, the tumor volume of the nude mice is photographed and recorded for 9 times, and the average value of the body weight of each group of nude mice is counted and made into a line graph, as shown in fig. 6. After the nude mice are inoculated with the breast cancer cells, the tumor volume of the nude mice in the infrared light irradiation group is increased in a geometric form, but is smaller than that of the nude mice in the blank group; the volume growth speed of the nude mouse tumor with the doxorubicin copper sulfide microneedle administration group is slow; the tumor volume of the doxorubicin copper sulfide nano microneedle administration and infrared light irradiation group nude mice is obviously smaller than that of the other three groups of nude mice, and the tumor of the group of nude mice is controlled. The result shows that the doxorubicin copper sulfide nanoparticle microneedle administration and infrared irradiation can well inhibit the growth of the breast cancer transplantation tumor of a nude mouse.

Test example 2 evaluation of penetration of microneedle patch into skin in vitro

Since breast cancer causes local thickening and hardening of the skin, a microneedle patch having better toughness and strength has a better effect. The surface condition of the mouse skin was observed after pressing the skin of the isolated mouse for 5min with a 200g weight on 3 microneedle patches A, B and C prepared according to the method provided by the present invention, wherein the mass ratio of the first polymer material to the second polymer material in the polymer materials of the 3 microneedle patches A, B and C was 3:10, 1:2 and 3:5, respectively. As shown in fig. 7, 3 kinds of microneedle patches were able to penetrate the skin. Therefore, the microneedle prepared by the invention has enough mechanical strength to penetrate into the skin and strong penetrating performance.

Test example 3 evaluation of safety of microneedle patch

The safety evaluation of the microneedles was performed by studying the body weight change of nude mice. The body weight change results of the nude mice in the blank microneedle group, the administration microneedle group, the blank microneedle + illumination group, and the administration microneedle + illumination group are shown in fig. 8. It can be seen that the weight of the nude mice inoculated with the breast cancer transplantable tumor tends to increase, and the quality of the four groups of nude mice is stabilized at 18-19 g after 24 days. The positive control group was not much different from the negative control group. The difference between the groups is not obvious and there is no rule to say, so there is no statistical difference in the effect of the transdermal administration of the doxorubicin-copper sulfide nanoparticle on the body weight of the nude mice. The microneedle has small systemic effect, high safety and no influence on the change of body weight of nude mice.

In conclusion, the embodiment of the invention combines doxorubicin and copper sulfide as a preparation and treats tumors by adopting a transdermal administration mode, so as to improve the treatment effect of breast cancer and reduce toxic and side effects. The copper sulfide nano-particles have the characteristics of no toxicity, uniform dispersion and high photothermal effect, have better tumor treatment effect, and have the specific mechanism of locally heating tumor parts under the action of near-infrared laser so as to kill tumor cells. The preparation method provided by the embodiment of the invention is simple in process and convenient to prepare. In addition, the microneedle patch prepared by the preparation method provided by the embodiment of the invention has strong mechanical strength and strong skin penetrating power.

In addition, it should be understood that the foregoing embodiments are merely exemplary illustrations of the present invention, and the technical solutions of the respective embodiments can be arbitrarily combined and collocated without conflict, contradiction and conflict between technical features.

Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims

1. A method of manufacturing a microneedle patch, comprising:

dissolving a high molecular material in water to obtain a high molecular polymer aqueous solution;

adding doxorubicin and copper sulfide nanoparticles into the high-molecular polymer aqueous solution to obtain a microneedle matrix solution;

placing the microneedle matrix solution into a microneedle female die, centrifuging and curing to obtain a microneedle matrix layer; and

and attaching the substrate layer to the microneedle matrix layer to obtain the microneedle patch.

2. The production method according to claim 1, wherein the polymer material includes a first polymer material and a second polymer material; the first high polymer material comprises one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid and chitosan, and the second high polymer material comprises one of monomer polymer or copolymer of carbopol, vinyl pyrrolidone and derivatives thereof.

3. The method according to claim 2, wherein the mass ratio of the first polymer material to the second polymer material is: (1-3) and (3-10).

4. The preparation method according to claim 1, wherein the mass ratio of the doxorubicin, the copper sulfide nanoparticles and the high molecular material is: (0.05-0.1) and (0.3-0.5) 1.

5. The method according to claim 1, further comprising a copper sulfide nanoparticle preparation step, wherein the copper sulfide nanoparticle preparation step specifically comprises:

adding sodium citrate and copper chloride into deionized water and stirring to obtain an initial solution;

adding a sodium sulfide aqueous solution into the initial solution and carrying out high-temperature oscillation to obtain a transition solution; and

and adding a sodium chloride ethanol aqueous solution into the transition solution at a low temperature, centrifuging, removing a supernatant, and washing to obtain the copper sulfide nano-particles.

6. The preparation method according to claim 5, wherein the mass ratio of the sodium citrate to the copper chloride is 1 (1.0-1.2); the mass ratio of sodium sulfide in the sodium sulfide aqueous solution is 20-26%.

7. The method according to any one of claims 1 to 6, further comprising a microneedle negative mold preparation step, the microneedle negative mold preparation step specifically comprising:

in a vacuum environment, the mass ratio is (0.5-3): (5-20) mixing the polydimethylsiloxane curing agent with the prepolymer to obtain a polydimethylsiloxane mixed solution; and

and casting the polydimethylsiloxane mixed solution on a male mold of a metal microneedle body and curing to obtain the microneedle female mold.

8. A microneedle patch, comprising:

a base layer; and

a microneedle matrix layer disposed on the base layer;

wherein, the microneedle matrix layer comprises a high molecular material, doxorubicin and copper sulfide nanoparticles.

9. The microneedle patch according to claim 8, wherein the mass ratio of the doxorubicin, the copper sulfide nanoparticles and the polymer material is: (0.05-0.1), (0.3-0.5) and (1).

10. A microneedle patch according to claim 8 or 9, wherein the polymer material comprises a first polymer material and a second polymer material; the mass ratio of the first high polymer material to the second high polymer material is as follows: (1-3) and (3-10); the first high polymer material comprises one of polyvinyl alcohol, polylactic acid-glycolic acid copolymer, polylactic acid, hyaluronic acid and chitosan, and the second high polymer material comprises one of monomer polymer or copolymer of carbopol, vinyl pyrrolidone and derivatives thereof.