TW201531308A - Ultrasound microbubble contrast agent for external use - Google Patents
Ultrasound microbubble contrast agent for external use Download PDFInfo
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本發明是關於一種外用超音波對比劑,特別是針對外用型微氣泡對比劑與其在化學藥物傳輸之應用。 The present invention relates to a topical ultrasonic contrast agent, particularly to a topical microbubble contrast agent and its use in chemical drug delivery.
對於大部分的化學品或藥物經皮吸收(transcutaneous absorption)的效率來說,皮膚障蔽功能(skin barrier function)為影響有效成分穿透皮膚最主要的關鍵因素,而擔任此障蔽功能者則是皮膚最外層的表皮層(厚度約10~15μm)。表皮層可分為最外層的角質層及其他活性表皮層,表皮穿透常受限於表皮層內的角質層所形成之屏障,導致藥物穿透不易。由於角質層的結構非常緻密,因此若分子量大於500道爾頓(Dalton)以上則不易穿透。另外由於脂質雙層是重要的藥物穿透路徑,親油性的非離子態的藥物原則上比親水性離子態的更容易穿透皮膚。 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.
為了克服經皮吸收受限於表皮層障礙的問題,本發明採 用物理性促進方法,以超音波方式搭配微氣泡對比劑協助藥物或化學品穿透皮膚,其優勢在於作用速度快,皮膚結構僅些微改變,皮膚恢復正常狀態的速度也較快。相較於傳統超音波對比劑須經由血管注射方式注入到生物體內,本案之微氣泡對比劑僅需施用至生物體表,無需要注射到體內,施用安全性高,適於美容、醫療或研究領域等。 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.
本發明提供一種外用型微氣泡對比劑,其使用時係塗覆 於生物體之體表,而不需採用習知的注射方式來使用。所述的外用型微氣泡對比劑,其介質可為水相或膠狀形態,並包含有特定濃度、粒徑大小之微氣泡。微氣泡材質可以是白蛋白、微脂體、聚合物、前述材質之共聚物或混合物或以上各者之組合。其可以直接施用於體表來使用,也可以搭配不同應用範圍之力學振盪能量來加以使用,藉由該力學振盪波動使得該些微氣泡經振盪能量之壓力誘導而生成脹與縮的一連串過程而使該些微氣泡破裂(destruction),進而產生能量及震波(shock wave)導致細胞或組織的細微破損,以加強搭配使用之各種化學物質的吸收效果。常見的力學振盪能量源可以是聲學能量或光學能量,例如是超音波或雷射光。所述微氣泡破裂產生之能量及震波對體表接觸部位之皮膚或黏膜造成微量且可回復的破損,藉此增加在該部位之化學物質之經皮吸收,因此可廣泛運用在醫療或美容等領域。舉例而言,可協助手術後之外用疼痛藥物吸收、加強各種美容美體保養成分之吸收等。 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.
本發明提供一種外用型微氣泡對比劑,至少包括一液態介質以及多個分散於該液態介質中之多個微氣泡。所述多個微氣泡由微氣泡材料組成,所述微氣泡對比劑是透過超音波震盪含葡萄糖的所述液態介質與所述微氣泡材料的混合液製備而成,所述微氣泡的平均顆粒直徑是透過所述微氣泡材料含量、所述液態介質葡萄糖的含量比例來控制。 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.
根據本發明之實施例,所述液態介質為等張生理食鹽 水,所述微氣泡材料為白蛋白、聚合物、微脂體、其共聚物或其組合。 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.
根據本發明之實施例,所述液態介質為等張生理食鹽水,所述微氣泡材料為人類血清白蛋白。。 According to an embodiment of the invention, the liquid medium is isotonic saline, and the microbubble material is human serum albumin. .
根據本發明之實施例,該些微氣泡粒徑介於0.5微米至3.7微米之間。 According to an embodiment of the invention, the microbubbles have a particle size between 0.5 microns and 3.7 microns.
本發明提供一種強化化學物質於體表的局部區域之滲透吸收的方法,該方法包含將前述微氣泡對比劑外用施用至體表的局部區域。將一治療有效劑量之化學物質施用至該局部區域。然後將一力學振盪源產生裝置直接接觸於該微氣泡組成物與該化學物質施用過的該局部區域,藉由該力學振盪源產生裝置產生之力學波,使該微氣泡對比劑中的該些微氣泡,受力學波作用而振盪擠壓,而增進該化學物質於該局部區域穿透過體表之滲透吸收。 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.
根據本發明之實施例,其中施用微氣泡對比劑之步驟與施用化學物質之步驟是分別施用,並非同時施用。 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.
根據本發明之實施例,其中該力學振盪源產生裝置包括一超音波裝置與/或一雷射光裝置。 According to an embodiment of the invention, the mechanical oscillation source generating device comprises an ultrasonic device and/or a laser device.
本發明提供一種強化化學物質於體表的局部區域之滲透吸收的方法,該方法之步驟一是將前述微氣泡對比劑外用施用至體表的局部區域,藉由該外力作用使該微氣泡對比劑中的該些微氣泡受力擠壓破裂,而增進該化學物質於該局部區域穿透過體表之滲透吸收,步驟二是將一治療有效劑量之化學物質施用至該局部區域以利其被吸收。 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. .
基於上述,本發明所提供之外用型微氣泡對比劑可安全有效地加強施用部位之化學物質之滲透吸收,且避免對比劑注入體內所可能導致之過敏風險。 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.
為讓本發明之上述特徵和優點能更明顯易懂,下文特列舉實施例配合所附圖解作詳細說明如下。 The above-described features and advantages of the present invention will be more apparent from the following description.
10‧‧‧仿體 10‧‧‧Faux
20、400‧‧‧超音波探頭 20,400‧‧‧Ultrasonic probe
30‧‧‧灌流區 30‧‧‧Irrigation area
35‧‧‧傳導膠 35‧‧‧Transmission adhesive
100‧‧‧培養皿 100‧‧‧ Petri dishes
200‧‧‧細胞 200‧‧‧ cells
300‧‧‧微氣泡對比劑 300‧‧‧Microbubble contrast agent
500‧‧‧超音波傳導膠 500‧‧‧Supersonic conductive adhesive
MB‧‧‧微氣泡 MB‧‧‧microbubbles
圖1為依據本發明一實施例之微氣泡對比劑搭配超音波施打之基因轉殖架構示意圖。 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.
圖2為依據本發明實施例以三種不同粒徑微氣泡對比劑與同樣的超音波條件來進行綠螢光基因轉殖表現量量化結果之條狀圖。 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.
圖3所示乃是圖2實施例以螢光顯微鏡觀察綠螢光基因轉殖表 現結果。 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.
圖4為依據本發明另一實施例之經皮穿透實驗系統結構示意圖。 4 is a schematic view showing the structure of a percutaneous penetration experiment system according to another embodiment of the present invention.
圖5為依據本發明實施例之經皮穿透實驗的穿透深度圖。 Figure 5 is a penetration depth map of a percutaneous penetration experiment in accordance with an embodiment of the present invention.
圖6為依據本發明實施例之各組經皮穿透實驗的穿透深度量化圖。 Figure 6 is a quantification of penetration depth for each group of percutaneous penetration experiments in accordance with an embodiment of the present invention.
圖7為依據本發明又一實施例之各組經皮穿透實驗的穿透深度量化圖。 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.
圖8A為超音波能量1W/cm2之染劑於豬皮穿透實驗的穿透深度放大100倍之顯微影像。 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.
圖8B為超音波能量1W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。 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.
圖9A為超音波能量2W/cm2之染劑於豬皮穿透實驗的穿透深度放大100倍之顯微影像。 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.
圖9B為超音波能量2W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。 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.
圖10A為超音波能量3W/cm2之染劑於豬皮穿透實驗的穿透深度放大100倍之顯微影像。 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.
圖10B為超音波能量3W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。 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.
本發明之微氣泡對比劑可為水相形態或膠狀形態,其中 有特定濃度、粒徑大小之微氣泡,而依其所含微氣泡之材質可分為大致三類:白蛋白微氣泡、微脂體微氣泡、或是聚合物微氣泡。 微氣泡對比劑所含之微氣泡具有穩定包覆的球殼,可被用來加強超音波反射的散射信號。微氣泡對比劑在不同的超音波強度能量作用之下,,將可加強化學物質對於超音波作用部位的滲透深度(亦即吸收效率)與/或穿透量(亦即吸收量)。 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.
本發明之微氣泡對比劑,可選擇白蛋白作為為唯一主原 料,或使用白蛋白當主原料同時搭配不同含量的葡萄糖,透過同一製程但是搭配不同原料的比例,即可製作出所需粒徑之微氣泡。本發明主要透過原料比例之調整,免除了過去必需搭配不同製程或多次離心過濾等繁瑣的物理方法,以精簡的製程達到製作不同粒徑之外用微氣泡。 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.
本發明之微氣泡對比劑可具有不同粒徑大小之微氣泡。 不同粒徑大小之微氣泡結合同一超音波能量會有不同程度之加強效果。本發明驗證不同粒徑之微氣泡搭配不同能量的超音波,能夠達到不同深度或濃度之穿透效果,故可針對選定的作用部位及預期的效果對象來加以調整。 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.
微氣泡對比劑在適當的聲場能量作用下,微氣泡對比劑 會承受很大的擠壓以及舒張,導致微氣泡對比劑破裂成碎片進而發生線性散射與穴蝕效應。穴蝕效應所產生之震波會導致細胞膜擾動而增加其通透性。根據研究顯示,在高聲場的情況下微氣泡 對比劑所產生的穴蝕效應,可以加強微循環滲透(microvascular leakages)、免疫細胞的滲透(inflammatory cell infiltrations)、溶解血栓(hemolysis)、甚或造成微血管破裂(capillary ruptures)等情況。 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.
本發明所提供之外用型微氣泡對比劑,乃係用於塗覆或 塗抹至生物體體表面之特定區域(亦即外用),該外用型微氣泡對比劑可加強生物體體表部位對於混合在該外用型微氣泡對比劑中的吸收效果。 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.
相較於一般所使用注入於體內血液循環系統的微氣泡對 比劑來說,本發明是將微氣泡對比劑設計成為介於超音波探頭與超音波作用部位(體表局部區域,如面部、關節部位或耳膜等)中間的介質,也就是說微氣泡係穩定存在於微氣泡對比劑中並且直接與超音波探頭接觸,在超音波能量誘導下產生穴蝕效應,進而加強體表部位對於其所搭配的化學物質的吸收與利用。此外,由於與本發明的微氣泡對比劑混合使用的化學物質並非包覆於微氣泡內,因此,它可以與本發明之微氣泡對比劑混合使用或是分開使用,亦即該等化學物質可依不同先後順序施用或塗覆至體表。 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.
本發明之外用型微氣泡對比劑,可以針對微氣泡濃度以 及介質張力來進行設計調整,而使微氣泡對比劑成為直接接觸超音波探頭的外用製劑。微氣泡對比劑所使用之介質可以為水相或膠狀形態,並且在搭配特定濃度之微氣泡後仍可具備有效之聲學傳導特性。微氣泡對比劑所使用之微氣泡材質可以是白蛋白、微脂體、聚合物、前述材質之共聚物或混合物,或以上各者之組合。 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.
本發明更提供一種外用型超音波微氣泡組成物之用途,其係外用施用於體表的局部區域以促進具藥效之化學物質穿透該局部區域的皮膚或黏膜,強化其滲透吸收。該外用型超音波微氣泡組成物包括至少一介質與分散於該介質中之多個微氣泡,其中該介質是液狀、膠狀形態,該些微氣泡材質選自白蛋白、聚合物或微脂體、前述材質之共聚物或混合物,或以上各者之組合。 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.
本發明更提供一種微氣泡組成物,所述微氣泡組成物可 以直接施用於體表的局部區域,僅須直接塗抹或稍加外力按摩擠壓,也會產生的穴蝕效應而使微氣泡破裂,進而促進具藥效之化學物質穿透該局部區域之皮膚或黏膜,強化該物質的滲透吸收。 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.
以下實施例係以白蛋白微氣泡對比劑為例,但本發明之微氣泡對比劑並不僅限於下列實施例之內容。 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.
實施例 Example
外用型微氣泡製劑之製作步驟:方法一:水相微氣泡對比劑製作:將生理食鹽水與1.2wt%人類血清白蛋白(human serum albumin,HSA,購自Octapharma,Vienna,Austria)均勻混合成10毫升之溶液並充入C3F8氣體後,以細胞粉碎儀振盪2分鐘以製成微氣泡對比劑,其在振盪過程中會形成以白蛋白為球殼包覆C3F8的微氣泡。將震盪完成的微氣泡對比劑分裝至微量離心管中,置於微量離心機中進行分離(轉速:1200rpm(128.7g)、時間:2分鐘),抽取下清液並加入適量的生理食鹽水,可保存於4℃冰箱中。在本 方法所製備的對比劑所包含之微氣泡濃度為4×108~2x109顆/ml而微氣泡粒徑分佈為0.5~3.7μm。 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.
A劑:以生理食鹽水為介質,將各種商用脂質球殼之微氣泡(包括磷脂類微氣泡SonoVue®(購自Bracco Diagnostics,Milan,Italy)或Definity(購自Lantheus Medical Imaging Inc,Billerica,MA)或Targestar(購自Targeson,La Jolla,CA)或前述自製之白蛋白球殼微氣泡對比劑的濃度,皆調整成1×109~2×109顆/ml(微氣泡原液)。 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).
B劑:配製搭配運用之化學物、生物分子或藥品等,所欲搭配之物質須調配為液狀、乳狀或膠狀,並使其與人體細胞成等張而pH值=7.4。化學物、生物分子或藥品可以為例如止痛藥雙氯芬酸、熊果素、維他命C磷酸鎂鹽、美白成分九胜肽、健大黴素或糖皮質激素等物質。 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劑:以B劑作為稀釋劑而將A劑進行2~1000倍稀釋混合後所得到之混合組成物,將其塗抹應用於生物體體表。其中效果特佳組為以B劑將A劑作2~40倍稀釋,較佳組為以B劑將A劑作30~150倍稀釋,或者也可以用B劑將A劑作100~1000倍稀釋。根據實驗,施用於皮膚時,10倍稀釋效果為最佳,但其他稀釋濃度也有效力,但效果不一。故應視施用區域而調整。一般而言,外用對比劑中微氣泡濃度較佳為約2x106~2 x108顆/ml。 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.
一般而言,可以將超音波探頭直接與生物體體表塗抹之C劑接觸,來進行局部超音波施打,能量為功率0.1~5W/cm2,而機 械指數(MI)<1.9。此外,與C劑作用之超音波能量,可以使用其他可產生力學振盪效應能量形式之裝置進行替代或合併使用,例如,治療用的雷射光束,並非全然受限於超音波的裝置使用。 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.
而本發明製備微氣泡對比劑之方式僅需控制原料成分比例,執行同一製程即可製備得到不同粒徑之微氣泡對比劑,製程精簡單純,可有效降低生產成本。 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.
方法二:特定粒徑微氣泡對比劑製作步驟:類似於上述方法一自製之白蛋白球殼微氣泡對比劑之製作方式,以生理食鹽水(或含有葡萄糖之生理食鹽水)搭配不同濃度人類血清白蛋白來製備出不同粒徑微氣泡(參見表1)的微氣泡對比劑。 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).
本案透過不同實驗驗證,可以利用控制原料配方比例來製作不同粒徑的白蛋白微氣泡對比劑。使用原料主要為白蛋白、生理食鹽水與/或葡萄糖。將白蛋白與葡萄糖混和或溶解於生理食鹽水中,並依照下列表1所列舉成分比例調配,即可製備得到所需粒徑大小之微氣泡對比劑。以下各例皆使用重量百分比。 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.
欲製備白蛋白微氣泡對比劑粒徑為0.5-1μm範圍間,使用原料比例為0.5-1wt%的白蛋白。欲製備白蛋白微氣泡對比劑粒徑為1-1.5μm間,使用原料比例為1-1.5wt%白蛋白、1.32wt%白蛋白及4-6wt%葡萄糖或0.5-1wt%白蛋白及8-12wt%葡萄糖。欲製備白蛋白微氣泡對比劑粒徑為1.5-2μm間,使用原料比例為1.32wt%白蛋白及8-17wt%葡萄糖或4.5-5.2wt%白蛋白及43-47 wt%葡萄糖。欲製備白蛋白微氣泡對比劑粒徑為2-2.5μm間,原料比例為1.32wt%白蛋白及18-32wt%葡萄糖、1.8-2.2wt%白蛋白及8-12wt%葡萄糖、3.3-3.7wt%白蛋白及43-47wt%葡萄糖、3.3-3.7wt%白蛋白及8-12wt%葡萄糖或1.8-2.2wt%白蛋白及43-47wt%葡萄糖。欲製備白蛋白微氣泡對比劑粒徑落於2.5-3μm間,使用原料比例為1.8-5.2wt%白蛋白、1.32wt%白蛋白及38-42wt%葡萄糖或4.8-5.2wt%白蛋白及8-12wt%葡萄糖。欲製備白蛋白微氣泡對比劑粒徑大小落於3-3.7μm間,使用原料比例為1.32wt%白蛋白及43-47wt%葡萄糖或0.5-1wt%白蛋白及43-47wt%葡萄糖。根據此原料比例製作成白蛋白微氣泡對比劑,可得相對應之粒徑大小的白蛋白微氣泡對比劑。 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.
將依表1比例配製的生理食鹽水(或含有葡萄糖之生理食鹽水)與白蛋白溶液混合後。將超音波震盪儀(設定20kHz)的探頭直接接觸混合物施以超音波能量震盪100-140秒左右,即可製成特定粒徑之微氣泡對比劑(微氣泡原液),其粒徑範圍參見表1最左欄。 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.
前述自製之白蛋白球殼微氣泡對比劑所含的微氣泡濃度為2×109顆/ml(原液),可分裝或不分裝冰存於4℃環境中。 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.
本實施例中根據表1選用其中三種比例製作出微泡直徑(粒徑)範圍0.5-1μm、1-1.5μm及3-3.5μm之白蛋白微氣泡對比劑,微氣泡對比劑結合超音波進行實驗,觀測是否提升基因轉殖效率。如圖1所示,本實施例使用HEI-OC1細胞進行綠螢光基因轉殖,細胞200單層培養於培養皿100中,置入微氣泡對比劑300使微氣泡MB與細胞200接觸。於培養皿100下方放置超音波探頭400,而在探頭400與培養100皿之間塗覆一層超音波傳導膠500,而施打超音波能量(超音波能量為功率0.1-5W/cm2),以改善細胞通透度。若作用後殘存沒破的氣泡仍停留在皮膚上,則洗掉殘存的微氣泡MB之後,讓細胞200接觸包覆帶綠螢光基因質體的微脂體進行基因轉殖,而於24小時後觀察綠螢光基因轉殖之效率。圖2為綠螢光基因轉殖表現量量化結果之條狀圖,圖3所示乃是螢光顯微鏡觀察綠螢光基因轉殖表現結果。如圖2-3所示,不同粒徑大小之微氣泡對比劑結合相同能量超音波對於基因轉殖效率之提升是有所差異的,於此實施例中可觀察到,較大粒徑之微氣泡對比劑能更有效提升基因轉殖效率。因此,可以根據不同需求,選擇進行基因轉殖較有利之粒徑大小。 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.
經皮穿透實驗流程 Percutaneous penetration test procedure
圖4為依據本發明的實施例之仿體穿透實驗系統結構示意圖。先以濃度為0.3wt%之洋菜膠製作仿體10,以利用仿體來模擬人體皮膚並進行穿透實驗。超音波探頭20與仿體10距離為約5mm。傳導膠35係置於探頭20上以使得傳導膠35與仿體10距離為約3mm。請參見圖4,灌流區30係置於仿體10上方而傳導膠35則位於灌流區30上方。傳導膠35填於灌流區30與仿體10間的空隙防止染劑滲出。本發明所提供之微氣泡對比劑係用來作為傳導膠35,而所搭配施打之化學物質係置於灌流區30中。 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.
照前述之微氣泡製作方式,並將不同粒徑微氣泡搭配超音波來觀測其經皮穿透之深度,使用染劑Evans blue及與人體組織相似的仿體進行模擬保養成分及藥物於皮膚穿透深度的探討。 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.
超音波施打流程:將超音波基因暨藥物傳送儀(ST 2000V)探頭置放於仿體上方距離仿體5mm,將依文思藍(Evans blue)染劑與水配置0.25%溶液注入灌流區內,而超音波頭靜置於染劑中施打超音波(2W/cm2)進行穿透實驗,在靜置完預定時間後,即可利用顯微鏡觀察染劑之仿體穿透深度利用。將滲透完的仿體進行顯微鏡觀察量測穿透深度並利用Matlab軟體作影像處理分析並計算穿透深度。控制組乃是0.25wt%的依文思藍染劑食鹽水溶液。 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.
進行下列三組實驗並變化不同的參數,以找出對應於最佳之染劑穿透深度的條件:(1)僅含依文思藍染劑(以(C)來表示);(2)單純施打超音波組(以(U)來表示)、(3)粒徑1.4μm微氣泡對比劑組(U+1.4μm)、(4)粒徑2.1μm微氣泡對比劑組(U+2.1μm)以及 (5)粒徑3.5μm微氣泡對比劑組(U+3.5μm);搭配超音波能量為1W/cm2、2W/cm2及3W/cm2之不同條件。利用顯微鏡來觀察仿體穿透深度並利用計算程式MATLAB來計算穿透深度而得到量化結果,如圖5-6所示。圖5為依據本實施例之之各組經洋菜仿體穿透實驗的穿透深度及MATLAB分析後之結果圖。結果發現,在相同超音波能量下,含有粒徑較大的微氣泡的微氣泡對比劑,可提供較深的染劑穿透效果。 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.
接著,改變搭配的超音波能量條件,來探討其對於穿透深度之影響。 Next, change the collocation energy condition of the collocation to explore its effect on penetration depth.
進行下列三組實驗並變化不同的參數,以找出對應於最佳之染劑穿透深度的條件:搭配超音波能量為1W/cm2、2W/cm2及3W/cm2之不同條件,結合粒徑1.4μm微氣泡對比劑組、粒徑2.1μm微氣泡對比劑組及粒徑3.5μm微氣泡對比劑組(10倍稀釋,微氣泡濃度2×108顆/ml),靜置時間為1分鐘。圖6為依據實施例之各組經洋菜仿體穿透實驗的穿透深度量化圖。 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.
在另一實驗中,將灌流區放置於厚度約2mm豬皮上進行經皮穿透實驗,其之實驗方式係與仿體穿透實驗相似,搭配超音波能量為1W/cm2、2W/cm2及3W/cm2之不同條件,結合粒徑1.4μm微氣泡對比劑組、粒徑2.1μm微氣泡對比劑組及粒徑3.5μm微氣泡對比劑組,來探討其對於穿透深度之影響。圖七為依據實施例之各組經豬皮染劑穿透實驗的穿透深度量化圖。其之實驗結果如圖8A-10B所示,圖8A為超音波能量1W/cm2之染劑於豬皮 穿透實驗的穿透深度放大100倍之顯微影像。圖8B為超音波能量1W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。圖9A為超音波能量2W/cm2之染劑於豬皮穿透實驗的穿透深度放大100倍之顯微影像。圖9B為超音波能量2W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。圖10A為超音波能量3W/cm2之染劑於豬皮穿透實驗的穿透深度放大100倍之顯微影像。圖10B為超音波能量3W/cm2之染劑於豬皮穿透實驗的穿透深度放大400倍之顯微影像。 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.
圖8A-10B為染劑放置於豬皮上進行冷凍切片之顯微影像,各組皆靜置15分鐘觀察顯微影像,可以看見隨著超音波能量變大,染劑穿透的深度就越深。而微氣泡對比劑微氣泡的粒徑越大時,染劑穿透的深度也越深。就施用超音波能量1W/cm2來說,不同粒徑組別的穿透深度雖然有差,但並差異並不明顯,而在施用超音波能量2W/cm2及3W/cm2時,不同粒徑組別的穿透深度就開始有了較大的差異,此部分與仿體的結果一致。 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.
從圖8A-10B中可以得知,控制組平均穿透深度為12.24μm,加上超音波有利於幫助染劑直接滲透穿越表皮層,故穿透深度明顯較控制組深。控制組施用超音波能量1W/cm2、2W/cm2及3W/cm2分別得到穿透深度為12.78μm、14.07μm、15.5μm。粒徑1.4μm微氣泡對比劑組施用超音波能量1W/cm2、2W/cm2及3W/cm2作用後的穿透平均深度分別為14.6μm、18.79μm、22.12μm。粒徑2.1μm微氣泡對比劑組施用超音波能量1W/cm2、 2W/cm2及3W/cm2作用後的穿透平均深度分別為16μm、22.55μm、27.28μm。粒徑3.5μm微氣泡對比劑組施用超音波能量1W/cm2、2W/cm2及3W/cm2作用後的穿透平均深度分別為19μm、25.02μm、30.61μm。 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.
從穿透實驗結果可知,超音波搭配本發明之微氣泡對比劑可以使得染劑穿透較深或穿透較為均勻。而相對於仿體,豬皮的穿透實驗,更顯示出本發明之微氣泡對比劑確實有強化化學物滲透(穿透)之功效。 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).
本發明的外用型微氣泡對比劑在使用時,需先將本發明微氣泡對比劑與稀釋劑的稀釋以約1:2至1:1000之比例稀釋,所述的稀釋劑可以是本發明之微氣泡對比劑本身所含之介質,更增加其比例;或以藥效成分本身(亦即,前述之該些化學物質)作為稀釋劑。此外,本發明的外用型微氣泡對比劑之介質並非不侷限於傳統液狀等張溶液。本發明的外用型微氣泡對比劑之微氣泡球殼材質可以是白蛋白、聚合物、微脂體、前述材質之共聚物或混合物,或以上各者之組合。 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.
外用型微氣泡對比劑中的微氣泡濃度範圍係介於2x106~2x108顆/ml之間。該微氣泡對比劑中微氣泡則應佔該微氣泡對比劑與介質之組合物總重量之0.5~2wt%(重量百分份),可有效傳遞聲波。 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.
本發明所搭配使用之超音波能量範圍係較佳地屬於非聚焦式的低能超音波,其能量只有MI=0.2~0.4的範圍,相較美國 FDA所規範之醫用超音波MI須低於1.9,眼科使用須低於0.2;本發明搭配使用之超音波能量範圍不但遠低於其所規範之MI=1.9,也未包含在眼科使用範圍內。本發明所使用之超音波能量範圍並不會產生局部的溫度變化,在上述之本發明實驗的操作過程中進行溫度監控,發現溫度差異只在正負0.1度之間,因此本發明搭配使用之超音波能量範圍與熱效應無關。 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.
雖然本發明之微氣泡組成物可以搭配超音波使用,但是本發明之微氣泡組成物同樣可以直接施用於體表的局部區域,僅須直接塗抹或施以外力按摩擠壓,也會產生的穴蝕效應而使微氣泡破裂,進而促進具藥效之化學物質穿透該局部區域之皮膚或黏膜,強化該些化學物質的滲透吸收。 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.
雖然本發明已以實施例揭露如上,然其並非用以限定本發明,任何所屬技術領域中具有通常知識者,在不脫離本發明之精神和範圍內,當可作些許之更動與潤飾,故本發明之保護範圍當視後附之申請專利範圍所界定者為基準。 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.
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