TW201531308A - Ultrasound microbubble contrast agent for external use - Google Patents

Abstract

An ultrasound microbubble contrast agent for external use is provided. The ultrasound microbubble contrast agent applied to the body surface externally can safely and efficiently enhance the permeation and absorption of the drug or chemicals through the skin of the body.

Description

External type microbubble contrast agent

The present invention relates to a topical ultrasonic contrast agent, particularly to a topical microbubble contrast agent and its use in chemical drug delivery.

For the efficiency of transdermal absorption of most chemicals or drugs, the skin barrier function is the most important factor affecting the penetration of active ingredients into the skin, and the person who acts as the barrier function is the skin. The outermost skin layer (thickness of about 10~15μm). The epidermis layer can be divided into the outermost stratum corneum and other active epidermal layers. The penetration of the epidermis is often limited by the barrier formed by the stratum corneum in the epidermal layer, which makes drug penetration difficult. Since the structure of the stratum corneum is very dense, it is not easy to penetrate if the molecular weight is more than 500 Daltons. In addition, since the lipid bilayer is an important drug penetration path, the lipophilic non-ionic drug is in principle more likely to penetrate the skin than the hydrophilic ionic state.

In order to overcome the problem that percutaneous absorption is limited by epidermal barrier, the present invention adopts the present invention. Using physical promotion methods, ultrasonic wave matching microbubble contrast agent assists drugs or chemicals to penetrate the skin. The advantage is that the action speed is fast, the skin structure changes only slightly, and the skin returns to normal state at a faster rate. Compared with the traditional ultrasonic contrast agent, it must be injected into the living body by vascular injection. The microbubble contrast agent in this case only needs to be applied to the living body table, no need to be injected into the body, and the application safety is high, and it is suitable for beauty, medical treatment or research. Fields, etc.

The invention provides a micro-bubble contrast agent for external use, which is coated during use. It is used on the body surface of the organism without the use of conventional injection methods. The external-type microbubble contrast agent may be in the form of an aqueous phase or a gelatinous form, and contains microbubbles having a specific concentration and a particle size. The microbubble material may be albumin, a liposome, a polymer, a copolymer or mixture of the foregoing materials, or a combination of the above. It can be directly applied to the body surface for use, or can be used with mechanical oscillation energy of different application ranges, and the mechanical oscillation fluctuation causes the microbubbles to be induced by the pressure of the oscillation energy to generate a series of processes of expansion and contraction. The microbubble is destructed, which in turn generates energy and shock waves that cause micro-damage of cells or tissues to enhance the absorption of various chemicals used in combination. A common source of mechanical oscillation energy can be acoustic energy or optical energy, such as ultrasound or laser light. The energy and the shock wave generated by the microbubble rupture cause a slight and reversible damage to the skin or mucous membrane at the contact portion of the body surface, thereby increasing the percutaneous absorption of the chemical substance at the site, and thus can be widely used in medical or cosmetic treatment, etc. field. For example, it can assist in the absorption of pain medications after surgery, and enhance the absorption of various beauty and body care ingredients.

The invention provides a topical microbubble contrast agent comprising at least a liquid medium and a plurality of microbubbles dispersed in the liquid medium. The plurality of microbubbles are composed of a microbubble material prepared by ultrasonically oscillating a mixture of the liquid medium containing glucose and the microbubble material, and the average particle of the microbubbles The diameter is controlled by the content of the microbubble material and the ratio of the content of the liquid medium glucose.

According to an embodiment of the invention, the liquid medium is an isotonic physiological salt Water, the microbubble material is albumin, a polymer, a liposome, a copolymer thereof, or a combination thereof.

According to an embodiment of the invention, the liquid medium is isotonic saline, and the microbubble material is human serum albumin. .

According to an embodiment of the invention, the microbubbles have a particle size between 0.5 microns and 3.7 microns.

The present invention provides a method of enhancing the osmotic absorption of a chemical substance in a localized area of a body surface, the method comprising applying the aforementioned microbubble contrast agent to a localized area of the body surface for external application. A therapeutically effective amount of the chemical is applied to the localized area. Then, a mechanical oscillation source generating device is directly in contact with the microbubble composition and the local region to which the chemical substance is applied, and the mechanical wave generated by the mechanical oscillation source generating device makes the microbubble contrast agent The bubble is oscillated and squeezed by the action of the mechanical wave, and the osmotic absorption of the chemical substance in the local area penetrates the body surface is enhanced.

According to an embodiment of the invention, the step of applying the microbubble contrast agent and the step of applying the chemical are separately applied, not simultaneously.

According to an embodiment of the invention, the mechanical oscillation source generating device comprises an ultrasonic device and/or a laser device.

The invention provides a method for enhancing the osmotic absorption of a chemical substance in a partial region of a body surface. The first step of the method is to externally apply the microbubble contrast agent to a local region of the body surface, and the microbubble is compared by the external force. The microbubbles in the agent are crushed by force, and the osmotic absorption of the chemical substance in the local area is enhanced, and the second step is to apply a therapeutically effective dose of the chemical substance to the local area to facilitate absorption. .

Based on the above, the external microbubble contrast agent provided by the present invention can safely and effectively enhance the osmotic absorption of the chemical substance at the application site, and avoid the risk of allergies which may be caused by the injection of the contrast agent into the body.

The above-described features and advantages of the present invention will be more apparent from the following description.

10‧‧‧Faux

20,400‧‧‧Ultrasonic probe

30‧‧‧Irrigation area

35‧‧‧Transmission adhesive

100‧‧‧ Petri dishes

200‧‧‧ cells

300‧‧‧Microbubble contrast agent

500‧‧‧Supersonic conductive adhesive

MB‧‧‧microbubbles

1 is a schematic diagram of a gene transfer architecture of a microbubble contrast agent combined with ultrasonic application according to an embodiment of the invention.

2 is a bar graph of quantification of green fluorescent gene gene transfer performance by three different particle size microbubble contrast agents and the same ultrasonic conditions according to an embodiment of the present invention.

Figure 3 shows the green fluorescent gene gene transfer table observed by a fluorescent microscope in the embodiment of Figure 2. The result is now.

4 is a schematic view showing the structure of a percutaneous penetration experiment system according to another embodiment of the present invention.

Figure 5 is a penetration depth map of a percutaneous penetration experiment in accordance with an embodiment of the present invention.

Figure 6 is a quantification of penetration depth for each group of percutaneous penetration experiments in accordance with an embodiment of the present invention.

Figure 7 is a quantification of penetration depth for each group of percutaneous penetration experiments in accordance with yet another embodiment of the present invention.

Fig. 8A is a microscopic image in which the penetration intensity of the ultrasonic energy 1 W/cm ² dyed in the pig skin penetration test is magnified 100 times.

Fig. 8B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 1 W/cm ² in the pig skin penetration test.

Fig. 9A is a microscopic image in which the penetration intensity of the ultrasonic energy of 2 W/cm ² is magnified 100 times in the penetration depth of the pig skin penetration test.

Fig. 9B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 2 W/cm ² in the pig skin penetration test.

Fig. 10A is a microscopic image showing a magnification of 100 times the penetration depth of the dye energy of 3 W/cm ² in the pig skin penetration test.

Fig. 10B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 3 W/cm ² in the pig skin penetration test.

The microbubble contrast agent of the present invention may be in the form of an aqueous phase or a gelatinous form, wherein There are microbubbles of a specific concentration and particle size, and the materials of the microbubbles contained therein can be classified into three main types: albumin microbubbles, microlipid microbubbles, or polymer microbubbles. The microbubbles contained in the microbubble contrast agent have a stably coated spherical shell that can be used to enhance the scattered signal of ultrasonic reflection. The microbubble contrast agent, under the action of different ultrasonic intensity energies, will enhance the depth of penetration (i.e., absorption efficiency) and/or the amount of penetration (i.e., absorption) of the chemical for the ultrasonically acting site.

The microbubble contrast agent of the invention can select albumin as the sole main source Material, or use albumin as the main raw material at the same time with different levels of glucose, through the same process but with the proportion of different raw materials, you can make the micro-bubbles of the required particle size. The invention mainly adjusts the proportion of the raw materials, and eliminates the cumbersome physical methods that have to be matched with different processes or multiple centrifugal filtrations in the past, and uses the streamlined process to produce microbubbles with different particle sizes.

The microbubble contrast agent of the present invention may have microbubbles of different particle sizes. Microbubbles of different particle sizes combined with the same ultrasonic energy will have different degrees of enhancement. The invention verifies that the microbubbles of different particle sizes are matched with the ultrasonic waves of different energies, and can achieve the penetration effects of different depths or concentrations, so that the selected action parts and the expected effect objects can be adjusted.

Microbubble contrast agent under the action of appropriate sound field energy, microbubble contrast agent Will undergo a lot of squeezing and relaxation, causing the microbubble contrast agent to break into pieces and then linear scattering and cavitation effects. The shock wave generated by the cavitation effect causes cell membrane disturbance and increases its permeability. According to research, microbubbles in the case of high sound fields The cavitation effects produced by contrast agents can enhance microvascular leakages, inflammatory cell infiltrations, hemolysis, or even microvascular ruptures.

The external microbubble contrast agent provided by the invention is used for coating or The topical microbubble contrast agent is applied to a specific area of the surface of the living body (that is, for external use), and the absorption effect of the body surface portion of the living body for mixing in the external type microbubble contrast agent can be enhanced.

Compared with the microbubble pair injected into the blood circulation system in the body In the case of a specific agent, the present invention designs the microbubble contrast agent into a medium interposed between the ultrasonic probe and the ultrasonic action portion (a partial region of the body surface such as a face, a joint portion or an eardrum, etc.), that is, a microbubble system. Stable in the microbubble contrast agent and directly in contact with the ultrasonic probe, the cavitation effect is generated under the induction of ultrasonic energy, thereby enhancing the absorption and utilization of the chemical substances on the body surface. In addition, since the chemical substance used in combination with the microbubble contrast agent of the present invention is not coated in the microbubbles, it can be used in combination with or separately from the microbubble contrast agent of the present invention, that is, the chemicals can be used. Apply or apply to the body surface in a different order.

The external microbubble contrast agent of the present invention can be used for microbubble concentration The medium tension is used for design adjustment, and the microbubble contrast agent becomes an external preparation for direct contact with the ultrasonic probe. The medium used for the microbubble contrast agent can be in the form of an aqueous phase or a gel, and can still have effective acoustic conduction properties after being combined with a specific concentration of microbubbles. The microbubble material used for the microbubble contrast agent may be albumin, a liposome, a polymer, a copolymer or a mixture of the foregoing materials, or a combination of the above.

The present invention further provides a use of a composition for external use of ultrasonic microbubbles, which is applied externally to a localized area of a body surface to promote the penetration of a medicinal chemical substance into the skin or mucous membrane of the local area to enhance its osmotic absorption. The external-type ultrasonic microbubble composition comprises at least one medium and a plurality of microbubbles dispersed in the medium, wherein the medium is in a liquid or gel form, and the microbubbles are selected from the group consisting of albumin, polymer or liposome. a copolymer or mixture of the foregoing materials, or a combination of the above.

The invention further provides a microbubble composition, wherein the microbubble composition can In the local area directly applied to the body surface, it is only necessary to directly apply or slightly massage the external force, and the cavitation effect may also be generated to break the microbubbles, thereby promoting the penetration of the medicinal chemical substance into the skin of the local area. Or mucous membranes to enhance the penetration of the substance.

The following examples are exemplified by albumin microbubble contrast agents, but the microbubble contrast agent of the present invention is not limited to the contents of the following examples.

Example

Preparation steps of external microbubble preparation: Method 1: Preparation of aqueous microbubble contrast agent: physiological saline is uniformly mixed with 1.2 wt% human serum albumin (HSA, purchased from Octapharma, Vienna, Austria). After 10 ml of the solution was filled with C ₃ F ₈ gas, it was shaken by a cell pulverizer for 2 minutes to prepare a microbubble contrast agent which formed a microcapsule-coated C ₃ F ₈ microparticle during the shaking process. bubble. The shock-completed microbubble contrast agent was dispensed into a microcentrifuge tube and placed in a microcentrifuge for separation (rotation speed: 1200 rpm (128.7 g), time: 2 minutes), and the supernatant was withdrawn and an appropriate amount of physiological saline was added. It can be stored in a refrigerator at 4 °C. The contrast agent prepared by the method has a microbubble concentration of 4×10 ⁸ to 2×10 ⁹ particles/ml and a microbubble particle size distribution of 0.5 to 3.7 μm.

Agent A: Microbubbles of various commercial lipid globules (including phospholipid microbubbles SonoVue® (available from Bracco Diagnostics, Milan, Italy) or Definity (purchased from Lantheus Medical Imaging Inc, Billerica, MA) using physiological saline as a medium. Or the concentration of the Tarstarstar (purchased from Targeson, La Jolla, CA) or the aforementioned self-made albumin spherical shell microbubble contrast agent was adjusted to 1 × 10 ⁹ ~ 2 × 10 ⁹ / ml (microbubble stock solution).

Agent B: Prepare chemicals, biomolecules or medicines that are used in combination with the substances to be used. The substances to be mixed must be formulated into liquid, milky or gelatinous form, and made into isotonic with human cells and pH = 7.4. The chemical, biomolecule or drug may be, for example, an analgesic diclofenac, an arbutin, a vitamin C magnesium phosphate, a whitening ingredient, a quintapeptide, a gentamicin or a glucocorticoid.

C agent: A mixed composition obtained by diluting and mixing A agent 2 to 1000 times with a B agent as a diluent, and applying it to a living body surface. The best effect group is to dilute the A agent by 2~40 times with the B agent, and the preferred group is to dilute the A agent by 30~150 times with the B agent, or the agent A can be used for 100~1000 times with the B agent. dilution. According to the experiment, the 10-fold dilution effect is optimal when applied to the skin, but other dilution concentrations are also effective, but the effects are different. Therefore, it should be adjusted according to the application area. In general, the concentration of microbubbles in the external contrast agent is preferably about 2 x 10 ⁶ to 2 x 10 ⁸ /ml.

In general, the ultrasonic probe can be directly contacted with the C agent applied to the body surface to perform local ultrasonic application with an energy of 0.1 to 5 W/cm ² and a mechanical index (MI) of <1.9. In addition, the ultrasonic energy acting with the C agent can be replaced or used in combination with other devices that produce a mechanical oscillation effect energy form, for example, a therapeutic laser beam, which is not entirely limited to ultrasonic devices.

The method for preparing the microbubble contrast agent of the invention only needs to control the ratio of the raw material components, and the microbubble contrast agent with different particle diameters can be prepared by performing the same process, and the process is simple and pure, which can effectively reduce the production cost.

Method 2: a specific particle size microbubble contrast agent production step: similar to the above method, a homemade albumin spherical shell microbubble contrast agent is prepared by using physiological saline (or physiological saline containing glucose) with different concentrations of human serum Albumin to prepare microbubble contrast agents of different particle size microbubbles (see Table 1).

The case is verified by different experiments, and the ratio of raw material formula can be controlled to make albumin microbubble contrast agent with different particle sizes. The raw materials used are mainly albumin, physiological saline and/or glucose. The microbubble contrast agent of the desired particle size can be prepared by mixing or dissolving albumin with glucose in physiological saline solution and formulating according to the proportions of the components listed in Table 1 below. The following examples all use weight percentages.

To prepare an albumin microbubble contrast agent having a particle size ranging from 0.5 to 1 μm, albumin having a raw material ratio of 0.5 to 1 wt% is used. The albumin microbubble contrast agent is prepared to have a particle size of 1-1.5 μm, using a raw material ratio of 1-1.5 wt% albumin, 1.32 wt% albumin, and 4-6 wt% glucose or 0.5-1 wt% albumin and 8- 12wt% glucose. The preparation of albumin microbubble contrast agent has a particle size of 1.5-2 μm, using a raw material ratio of 1.32 wt% albumin and 8-17 wt% glucose or 4.5-5.2 wt% albumin and 43-47. Gt% glucose. The preparation of albumin microbubble contrast agent has a particle size of 2-2.5 μm, the ratio of raw materials is 1.32 wt% albumin and 18-32 wt% glucose, 1.8-2.2 wt% albumin and 8-12 wt% glucose, 3.3-3.7 wt. % albumin and 43-47 wt% glucose, 3.3-3.7 wt% albumin and 8-12 wt% glucose or 1.8-2.2 wt% albumin and 43-47 wt% glucose. The preparation of albumin microbubble contrast agent particle size falls between 2.5-3 μm, using a raw material ratio of 1.8-5.2 wt% albumin, 1.32 wt% albumin and 38-42 wt% glucose or 4.8-5.2 wt% albumin and 8 -12 wt% glucose. The size of the albumin microbubble contrast agent to be prepared falls between 3-3.7 μm, and the ratio of the raw materials is 1.32% by weight of albumin and 43-47% by weight of glucose or 0.5-1% by weight of albumin and 43-47% by weight of glucose. According to the ratio of the raw materials, the albumin microbubble contrast agent is prepared, and the corresponding albumin microbubble contrast agent can be obtained.

The physiological saline (or physiological saline containing glucose) prepared according to the ratio of Table 1 is mixed with the albumin solution. The probe of the ultrasonic oscillator (set at 20 kHz) is directly contacted with the mixture and subjected to ultrasonic energy oscillation for about 100-140 seconds to prepare a microbubble contrast agent (microbubble stock solution) of a specific particle size. 1 leftmost column.

The self-made albumin spherical shell microbubble contrast agent contains a microbubble concentration of 2×10 ⁹ particles/ml (stock solution), and can be stored in a 4° C. environment with or without ice.

In this example, according to Table 1, three kinds of ratios of 0.5-1 μm, 1-1.5 μm and 3-3.5 μm of albumin microbubble contrast agent were prepared, and the microbubble contrast agent was combined with ultrasonic wave. Experiment to see if it improves the efficiency of gene transfer. As shown in Fig. 1, in this example, HEI-OC1 cells were used for green fluorescent gene transfer, and cell 200 was monolayer cultured in a petri dish 100, and a microbubble contrast agent 300 was placed to bring the microbubbles MB into contact with the cells 200. The ultrasonic probe 400 is placed under the culture dish 100, and a layer of ultrasonic conductive adhesive 500 is applied between the probe 400 and the culture dish 100, and the ultrasonic energy is applied (the ultrasonic energy is 0.1-5 W/cm ² ). To improve cell permeability. If the remaining unbroken bubbles remain on the skin after washing, the remaining microbubbles MB are washed away, and the cells 200 are exposed to the microlipids coated with the green fluorescent gene plastid for gene transfer, and 24 hours. After the observation of the efficiency of green fluorescent gene transfer. Fig. 2 is a bar graph showing the quantitative results of the green fluorescent gene gene transfer expression, and Fig. 3 is a fluorescence microscope for observing the green fluorescent gene gene transfer performance. As shown in Figure 2-3, microbubble contrast agents of different particle sizes combined with the same energy ultrasound have different improvements in gene transfer efficiency. In this example, a larger particle size can be observed. Bubble contrast agent can improve the efficiency of gene transfer more effectively. Therefore, according to different needs, it is possible to select a particle size that is advantageous for gene transfer.

Percutaneous penetration test procedure

4 is a schematic view showing the structure of a pseudo-penetration experiment system according to an embodiment of the present invention. The imitation 10 was first prepared with a concentration of 0.3% by weight of acacia gum to simulate human skin using a pseudotype and perform a penetration test. The distance between the ultrasonic probe 20 and the phantom 10 is about 5 mm. A conductive adhesive 35 is placed on the probe 20 such that the conductive adhesive 35 is at a distance of about 3 mm from the analog 10. Referring to Figure 4, the perfusion zone 30 is placed over the imitation 10 and the conductive adhesive 35 is positioned above the perfusion zone 30. The conductive glue 35 is filled in the gap between the perfusion zone 30 and the imitation body 10 to prevent the dye from oozing out. The microbubble contrast agent provided by the present invention is used as the conductive paste 35, and the chemical substance to be applied is placed in the perfusion zone 30.

According to the above-mentioned microbubble production method, the micro-bubbles of different particle sizes are combined with ultrasonic waves to observe the depth of percutaneous penetration, and the dyeing agent Evans blue and the similar human body tissue are used to simulate the maintenance components and drugs on the skin. Through depth discussion.

Ultrasonic application process: The ultrasonic gene and drug delivery device (ST 2000V) probe is placed on the imitation body 5mm from the imitation body, and the Evans blue dye and water distribution 0.25% solution is injected into the perfusion area. The ultrasonic head is statically placed in the dye to apply ultrasonic (2W/cm ² ) for the penetration test, and after the predetermined time has elapsed, the depth of the imitation penetration of the dye can be observed by the microscope. The infiltrated phantom was subjected to microscopic observation to measure the penetration depth and analyzed by Matlab software for image processing and the penetration depth was calculated. The control group was a 0.25 wt% aqueous solution of Evans blue dye.

Perform the following three sets of experiments and change the different parameters to find the conditions corresponding to the best penetration depth of the dye: (1) contain only Evans blue dye (indicated by (C)); (2) simple application Ultrasonic group (indicated by (U)), (3) Microbubble contrast agent group (U+1.4 μm) with particle size of 1.4 μm, (4) Microbubble contrast agent group (U+2.1 μm) with particle size of 2.1 μm And (5) a microbubble contrast agent group (U + 3.5 μm) having a particle diameter of 3.5 μm; and different conditions of the ultrasonic energy of 1 W/cm ² , 2 W/cm ² and 3 W/cm ² . Use a microscope to observe the penetration depth of the phantom and use the calculation program MATLAB to calculate the penetration depth to obtain quantified results, as shown in Figure 5-6. Fig. 5 is a graph showing the penetration depth and the results of MATLAB analysis of each group of the analogy penetration experiment of the amaranth according to the present embodiment. It was found that under the same ultrasonic energy, the microbubble contrast agent containing microbubbles with larger particle diameters can provide a deeper dye penetration effect.

Next, change the collocation energy condition of the collocation to explore its effect on penetration depth.

Perform the following three sets of experiments and change the different parameters to find the conditions corresponding to the best penetration depth of the dye: with different conditions of ultrasonic energy of 1W/cm ² , 2W/cm ² and 3W/cm ² , Combined with a particle size of 1.4 μm microbubble contrast agent group, a particle size of 2.1 μm microbubble contrast agent group and a particle size of 3.5 μm microbubble contrast agent group (10-fold dilution, microbubble concentration 2×10 ⁸ particles/ml), standing time It is 1 minute. Fig. 6 is a quantification map of penetration depth of each group of ananas parison penetration test according to the examples.

In another experiment, the perfusion area was placed on pig skin with a thickness of about 2 mm for percutaneous penetration experiments. The experimental method was similar to the imitation penetration experiment with an ultrasonic energy of 1 W/cm ² and 2 W/cm. ² and 3W/cm ² different conditions, combined with particle size 1.4μm microbubble contrast agent group, particle size 2.1μm microbubble contrast agent group and particle size 3.5μm microbubble contrast agent group to explore its effect on penetration depth . Figure 7 is a quantification of penetration depth of each group of pig skin dye penetration experiments according to the examples. The experimental results are shown in Figs. 8A-10B. Fig. 8A is a microscopic image in which the ultrasonic wave energy of 1 W/cm ² is magnified 100 times in the penetration depth of the pig skin penetration test. Fig. 8B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 1 W/cm ² in the pig skin penetration test. Fig. 9A is a microscopic image in which the penetration intensity of the ultrasonic energy of 2 W/cm ² is magnified 100 times in the penetration depth of the pig skin penetration test. Fig. 9B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 2 W/cm ² in the pig skin penetration test. Fig. 10A is a microscopic image showing a magnification of 100 times the penetration depth of the dye energy of 3 W/cm ² in the pig skin penetration test. Fig. 10B is a microscopic image of a 400-fold magnification of the penetration depth of the dye energy of 3 W/cm ² in the pig skin penetration test.

8A-10B are microscopic images of frozen sections placed on pig skin, and each group was allowed to stand for 15 minutes to observe the microscopic image. It can be seen that as the ultrasonic energy becomes larger, the depth of penetration of the dye is higher. deep. When the particle size of the microbubble contrast agent microbubble is larger, the penetration depth of the dye is deeper. As far as the application of ultrasonic energy 1W/cm ² is concerned, although the penetration depths of different particle size groups are poor, the difference is not obvious, but when the ultrasonic energy is applied at 2W/cm ² and 3W/cm ² , the difference is different. The penetration depth of the particle size group began to vary greatly, and this part is consistent with the result of the imitation.

It can be seen from Fig. 8A-10B that the average penetration depth of the control group is 12.24 μm, and the ultrasonic wave is beneficial to help the dye directly penetrate the skin layer, so the penetration depth is obviously deeper than that of the control group. The control group applied ultrasonic energy of 1 W/cm ² , 2 W/cm ² and 3 W/cm ² to obtain penetration depths of 12.78 μm, 14.07 μm, and 15.5 μm, respectively. The average penetration depth after application of the ultrasonic energy of 1 W/cm ² , 2 W/cm ² and 3 W/cm ^{2 in the} microbubble contrast agent group of the particle diameter of 1.4 μm was 14.6 μm, 18.79 μm, and 22.12 μm, respectively. The average penetration depth after application of the ultrasonic energy of 1 W/cm ² , 2 W/cm ² and 3 W/cm ^{2 in the} microbubble contrast agent group of the particle diameter of 2.1 μm was 16 μm, 22.55 μm, and 27.28 μm, respectively. The average penetration depth after application of the ultrasonic energy of 1 W/cm ² , 2 W/cm ² and 3 W/cm ^{2 in the} microbubble contrast agent group of the particle diameter of 3.5 μm was 19 μm, 25.02 μm, and 30.61 μm, respectively.

It can be seen from the results of the penetration test that the ultrasonic wave is matched with the microbubble contrast agent of the present invention to make the dye penetration deeper or more uniform. Compared with the imitation, the penetration test of the pig skin shows that the microbubble contrast agent of the present invention does have the effect of enhancing chemical penetration (penetration).

When the external-type microbubble contrast agent of the present invention is used, the dilution of the microbubble contrast agent of the present invention and the diluent is first diluted at a ratio of about 1:2 to 1:1000, and the diluent may be the present invention. The medium contained in the microbubble contrast agent itself is increased in proportion; or the medicinal ingredient itself (that is, the aforementioned chemicals) is used as a diluent. Further, the medium of the external-purpose microbubble contrast agent of the present invention is not limited to the conventional liquid isotonic solution. The microbubble spherical shell material of the external-purpose microbubble contrast agent of the present invention may be albumin, a polymer, a liposome, a copolymer or a mixture of the foregoing materials, or a combination of the above.

The microbubble concentration range of the external type microbubble contrast agent is between 2 x 10 ⁶ and 2 x 10 ⁸ /ml. The microbubbles in the microbubble contrast agent should account for 0.5 to 2 wt% (weight percent) of the total weight of the composition of the microbubble contrast agent and the medium, and can effectively transmit sound waves.

The ultrasonic energy range used in conjunction with the present invention is preferably a non-focusing low-energy ultrasonic wave, and its energy is only in the range of MI=0.2-0.4, compared with the United States. The medical supersonic MI specified by the FDA must be less than 1.9, and the ophthalmic use must be less than 0.2; the ultrasonic energy range used in conjunction with the present invention is not only far below the normative MI=1.9, nor is it included in the ophthalmic use range. . The ultrasonic energy range used in the present invention does not cause local temperature change, and temperature monitoring is performed during the operation of the above-described experiment of the present invention, and it is found that the temperature difference is only between plus and minus 0.1 degrees, so the present invention is used in combination with the super The range of sonic energy is independent of thermal effects.

Although the microbubble composition of the present invention can be used in combination with ultrasonic waves, the microbubble composition of the present invention can also be directly applied to a partial area of the body surface, and only needs to be directly applied or applied by external force massage, and a hole can be generated. The opaque effect causes the microbubbles to rupture, thereby promoting the penetration of the medicinal chemical into the skin or mucous membrane of the localized area and enhancing the osmotic absorption of the chemical substances.

Although the present invention has been disclosed in the above embodiments, it is not intended to limit the invention, and any one of ordinary skill in the art can make some modifications and refinements without departing from the spirit and scope of the invention. The scope of the invention is defined by the scope of the appended claims.

Claims (10)

A topical microbubble contrast agent comprising at least: a liquid medium and a plurality of microbubbles dispersed in the liquid medium, wherein the plurality of microbubbles are composed of a microbubble material, and the microbubble contrast agent is permeated Ultrasonic vibration is prepared by oscillating a mixture of the liquid medium containing glucose and the microbubble material, and an average particle diameter of the microbubbles is transmitted through the microbubble material content, the liquid medium glucose content, and the The ratio of the microbubble material content to the content of the liquid medium glucose is controlled. The microbubble contrast agent for external use according to claim 1, wherein the liquid medium is isotonic physiological saline, and the microbubble material is human serum albumin. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 0.5 to 1 μm, and the microbubble material has an albumin content of 0.5 to 1 wt%. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 1-1.5 μm, and the microbubble material content is 1-1.5 wt% albumin, the micro The bubble material content is 1.32 wt% albumin and the liquid medium glucose content is 3-7 wt% glucose or the microbubble material content is 0.5-1 wt% albumin and the liquid medium glucose content is 8-12 wt% glucose. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 1.5 to 2 μm, and the microbubble material content is 1.32 wt% albumin and the liquid medium glucose content is 8-17 wt% glucose or the microbubble material content is 4.8-5.2 wt% albumin and the liquid medium glucose content is 43-47 wt% glucose. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 2 to 2.5 μm, the microbubble material content is 1.32 wt% albumin, and the liquid medium is glucose. The content is 18-32 wt% glucose, the microbubble material content is 1.8-2.2 wt% albumin and the liquid medium glucose content is 8-12 wt% glucose, and the microbubble material content is 3.3-3.7 wt% The albumin concentration and the liquid medium glucose content is 43-47 wt% glucose, the microbubble material content is 3.3-3.7 wt% albumin and the liquid medium glucose content is 8-12 wt% glucose or the micro The bubble material content is 1.8-2.2 wt% albumin and the liquid medium glucose content is 43-47 wt% glucose. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 2.5 to 3 μm, and the microbubble material has a content of 1.8 to 5.2 wt% of albumin, the microbubbles. The material content is 1.32 wt% albumin and the liquid medium glucose content is 38-42 wt% glucose or the microbubble material content is 4.8-5.2 wt% albumin and the liquid medium glucose content is 8-12 wt%. glucose. The external-type microbubble contrast agent according to claim 1, wherein the microbubbles have an average particle diameter of 3-3.7 μm, the microbubble material content is 1.32 wt% albumin, and the liquid medium glucose The content is 43-47 wt% glucose or the microbubble material content is 0.5-1 wt% albumin and the liquid medium glucose The content is 43-47 wt% glucose. A method for enhancing osmotic absorption of a chemical substance in a partial region of a body surface, the method comprising: applying a topical microbubble contrast agent as described in claims 1 to 8 to a partial region of a body surface; An effective dose of the chemical substance is applied to the localized region; and a mechanical oscillation source generating device is directly in contact with the microbubble composition and the localized region to which the chemical substance is applied, and the mechanical wave generated by the mechanical oscillation source generating device The microbubbles in the microbubble contrast agent are oscillated and squeezed by the action of mechanical waves to enhance the permeation absorption of the chemical substance through the body surface in the local region. A method for enhancing osmotic absorption of a chemical substance in a partial region of a body surface, the method comprising: applying a topical microbubble contrast agent as described in claims 1 to 8 to a partial region of a body surface; An effective dose of the chemical substance is applied to the localized area; and an external force is directly in contact with the microbubble composition and the localized area to which the chemical substance is applied, and the microbubbles in the microbubble contrast agent are affected by the external force The force squeezes the rupture and enhances the osmotic absorption of the chemical through the body surface in the localized area.