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KR20170041375A - A Microneedle skin patch functionalized by Aptamer-coated carbon nanotube for early diagnosis of various diseases - Google Patents

A Microneedle skin patch functionalized by Aptamer-coated carbon nanotube for early diagnosis of various diseases Download PDF

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KR20170041375A
KR20170041375A KR1020150140741A KR20150140741A KR20170041375A KR 20170041375 A KR20170041375 A KR 20170041375A KR 1020150140741 A KR1020150140741 A KR 1020150140741A KR 20150140741 A KR20150140741 A KR 20150140741A KR 20170041375 A KR20170041375 A KR 20170041375A
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손인식
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/44Detecting, measuring or recording for evaluating the integumentary system, e.g. skin, hair or nails
    • A61B5/441Skin evaluation, e.g. for skin disorder diagnosis
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/0002Remote monitoring of patients using telemetry, e.g. transmission of vital signals via a communication network
    • AHUMAN NECESSITIES
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    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/05Detecting, measuring or recording for diagnosis by means of electric currents or magnetic fields; Measuring using microwaves or radio waves
    • A61B5/053Measuring electrical impedance or conductance of a portion of the body
    • A61B5/0531Measuring skin impedance
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/145Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue
    • A61B5/14507Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue specially adapted for measuring characteristics of body fluids other than blood
    • A61B5/1451Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue specially adapted for measuring characteristics of body fluids other than blood for interstitial fluid
    • A61B5/14514Measuring characteristics of blood in vivo, e.g. gas concentration or pH-value ; Measuring characteristics of body fluids or tissues, e.g. interstitial fluid or cerebral tissue specially adapted for measuring characteristics of body fluids other than blood for interstitial fluid using means for aiding extraction of interstitial fluid, e.g. microneedles or suction
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/68Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
    • A61B5/6801Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be attached to or worn on the body surface
    • A61B5/683Means for maintaining contact with the body
    • A61B5/6832Means for maintaining contact with the body using adhesives
    • A61B5/6833Adhesive patches
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61BDIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00Measuring for diagnostic purposes; Identification of persons
    • A61B5/68Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient
    • A61B5/6846Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive
    • A61B5/6847Arrangements of detecting, measuring or recording means, e.g. sensors, in relation to patient specially adapted to be brought in contact with an internal body part, i.e. invasive mounted on an invasive device
    • A61B5/685Microneedles

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Abstract

본 발명은 다양한 질병의 조기진단용 앱타머 코팅된 탄소 나노튜브로 기능화된 마이크로니들 스킨 패치에 관한 것으로, 본 발명의 패치는 피부내 세포간질에 존재하는 다양한 바이오마커가 이에 특이적으로 반응하는 앱타머와 결합했을 때 나타나는 전기전도도의 변화를 실시간으로 감지하여 이를 증폭시켜서 외부의 기기에 효과적으로 전송함으로써 다양한 질환을 조기에 진단할 수 있다.The present invention relates to a micro needle skin patch functionalized with an aptamer-coated carbon nanotube for early diagnosis of various diseases, and a patch of the present invention is characterized in that a variety of biomarkers existing in the subepithelial cell epilepsy are aptamers It is possible to diagnose various diseases at an early stage by effectively detecting the change of the electric conductivity when it is coupled with an external device in real time and amplifying it.

Description

다양한 질병의 조기진단용 앱타머 코팅된 탄소 나노튜브로 기능화된 마이크로니들 스킨 패치{A Microneedle skin patch functionalized by Aptamer-coated carbon nanotube for early diagnosis of various diseases}Technical Field [0001] The present invention relates to a micro-needle skin patch functionalized with an aptamer-coated carbon nanotube for early diagnosis of various diseases.

본 발명은 다양한 질병의 조기진단용 앱타머 코팅된 탄소 나노튜브로 기능화된 마이크로니들 스킨 패치에 관한 것이다.The present invention relates to a micro needle skin patch functionalized with an aptamer-coated carbon nanotube for early diagnosis of various diseases.

피부 질환은 오늘날 세계에서 주요 건강 관리 도전을 대표한다. 미국(국립암연구소, www.cancer.gov)에서 해마다 새로이 진단되는 백만 건 이상의 피부암과 더불어, 피부 질환을 예측 및 진단하는 것은 그것의 관리 측면에서 중요하다. 현재의 진단 방법은 시각적인 관찰 및 생검에 주로 의존한다. 그러나, 시각적인 관찰에 의존하는 검출 방법은 피부 상태 또는 질환을 진단하는데 반드시 효과적인 것은 아니며, 임상적인 징후가 나타날 때까지는 위험(risk) 또는 질환을 검출하지 못한다. 게다가, 생검과 같은 침습성 방법은 시험 대상에 대한 외상뿐만 아니라, 감염의 가능성도 증가시킨다. 또한 상기 방법은 안전하게 실시되기 위해서 의사에 의해 수행되어야 하며, 일반적으로 반응에 관여하는 세포인 피부 표면에 있는 풍부한 세포 샘플을 보통 제공하지 않는다.Skin diseases represent a major healthcare challenge in the world today. Along with more than one million new skin cancers diagnosed annually in the US (National Cancer Institute, www.cancer.gov), predicting and diagnosing skin disorders is important in its management. Current diagnostic methods rely mainly on visual observation and biopsy. However, detection methods that rely on visual observations are not necessarily effective in diagnosing a skin condition or disease, and do not detect a risk or disease until clinical manifestations occur. In addition, invasive methods such as biopsy increase the likelihood of infection as well as trauma to the test subject. In addition, the method should be performed by a physician in order to be safely performed, and usually does not provide a rich cell sample on the surface of the skin, which is generally a cell involved in the response.

따라서 피부 상태 및 질환을 진단 및 모니터링하는 비침습성 방법은 환자의 관리를 위한, 그리고 기존 및 새로운 치료제, 피부 관리 제품 및 피부 관리 섭생의 효능을 평가하기 위한 중요한 수단을 나타낸다. 게다가, 상기방법은 피부 질환의 발생에 대한 시험 대상의 유전적 소인뿐만 아니라, 시험 대상의 피부 상태를 기초로 하는 특이적인 유전적 변화에 관하여 중요한 정보를 제공할 수 있다. 상기 유전적 변화를 동정하는 것은 잠재적인 약물 타겟 및 예방적 조치를 동정하고 특정 치료제, 피부 관리 제품 또는 섭생에 대해 사람이 실제적으로 반응을 하는지를 결정하는데 중요할 수 있다. 뿐만 아니라, 검출 및 진단 방법은 그러한 치료, 제품 및 조치의 안전성을 평가하는데 중요하다.Non-invasive methods for diagnosing and monitoring skin conditions and diseases therefore represent an important means for the management of patients and for evaluating the efficacy of existing and new treatments, skin care products and skin care regimens. In addition, the method can provide important information about specific genetic changes based on the skin condition of the test subject, as well as the genetic susceptibility of the test subject to the occurrence of skin disease. Identifying the genetic alteration may be important in identifying potential drug targets and preventative measures and determining whether a person actually responds to a particular therapeutic agent, skin care product or regimen. In addition, detection and diagnostic methods are important in assessing the safety of such treatments, products and measures.

또한, 국소적인 피부질환은 물론 다양한 질병상태에서 피부의 물질의 조성이 바뀌는 것이 보고되고 있다. 지질, 구조단백질, 염증성물질, 핵산, 대사산물 등 여러물질이 질병상태에 따라 피부에서 다양하게 검출되는 것으로 알려져있다. 현재 아토피성 피부염, 흑색종, 피부의 세균성 염증 외에도 알츠하이머병, 파킨슨씨병, 유방암, 심혈관계 질환, 당뇨병, 약물 중독등의 다양한 질환에서 피부의 바이오마커 분석이 이루어지고 있다. 그러나, 대부분의 경우 대단히 침습적인 피부생검(Skin Biopsy)이 사용되고 있다. 비침습적 방법으로 Iontophoresis, Microdialysis, Tape stripping, Ultrasound, Microneedle등이 사용되고 있으나, 매우 효율은 낮은 편이다 (Paliwal et al., 2013 Diagnostic opportunities based on skin biomarkers. European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences 50:546-556). In addition, it has been reported that the composition of skin material changes in various disease states as well as in local skin diseases. It is known that various substances such as lipids, structural proteins, inflammatory substances, nucleic acids, metabolites and the like are variously detected in the skin depending on the disease state. In addition to atopic dermatitis, melanoma and bacterial inflammation of the skin, biomarker analysis of skin has been performed in various diseases such as Alzheimer's disease, Parkinson's disease, breast cancer, cardiovascular disease, diabetes, drug addiction and the like. However, in most cases, a very aggressive skin biopsy is being used. In the noninvasive method, Iontophoresis, Microdialysis, Tape stripping, Ultrasound, and Microneedle are used, but they are very low in efficiency (Paliwal et al., 2013 Diagnostic opportunities based on biomarkers. Federation for Pharmaceutical Sciences 50: 546-556).

[선행 특허 문헌][Prior Patent Literature]

대한민국 특허공개번호 제1020120006945호Korean Patent Publication No. 1020120006945

본 발명은 상기의 문제점을 해결하고 필요성에 의하여 안출된 것으로서 본 발명의 목적은 다양한 질환의 조기 진단에 이용하기 위한 패치를 제공하는 것이다. SUMMARY OF THE INVENTION The present invention has been made in view of the above problems, and it is an object of the present invention to provide a patch for use in early diagnosis of various diseases.

상기의 목적을 달성하기 위하여 본 발명은 마이크로니들(Microneedle)을 앱타머로 코팅된 탄소나노튜브 전계효과트랜지스터 (CNT-FET)로 기능화하여 피부 내 세포간질에 존재하는 바이오마커가 특이적으로 반응하는 앱타머와 결합했을 때 나타나는 전기전도도의 변화를 실시간으로 감지하여 이를 증폭시켜서 외부의 기기에 효과적으로 전송함으로써 질환을 조기에 진단할 수 있게 하는 진단용 스킨패치를 제공한다.In order to achieve the above object, the present invention provides an application in which a microneedle is functionalized with a carbon nanotube field effect transistor (CNT-FET) coated with an aptamer to specifically react with a biomarker existing in the cell epilepsy in the skin It provides a diagnostic skin patch that enables real-time detection of electrical conductivity changes when combined with a tamer, amplifies it, and effectively transmits it to external devices to diagnose the disease early.

본 발명의 일 구현예에 있어서, 상기 바이오마커는 단백질, 펩타이드, DNA 또는 RNA인 것이 바람직하나 이에 한정되지 아니한다.In an embodiment of the present invention, the biomarker is preferably a protein, a peptide, DNA, or RNA, but is not limited thereto.

본 발명의 일 구현예에 있어서, 상기 외부 기기는 스마트폰, 휴대폰, 또는 무선통신기기인 것이 바람직하나 이에 한정되지 아니한다.
In one embodiment of the present invention, the external device is a smart phone, a mobile phone, or a wireless communication device, but is not limited thereto.

이하 본 발명을 설명한다.Hereinafter, the present invention will be described.

앱타머로 coating된 탄소나노튜브 전계효과트랜지스터 (CNT-FET)로 기능화한 마이크로니들 스킨 패치를 이용해 각질층 아래 표피층에 존재하는 다양한 질병의 표지자 (Biomarkers)를 실시간으로 검출함으로써 다양한 질환의 조기 진단에 이용하는 것이 발명의 목적이다.Biomarkers of various diseases present in the epidermis layer below the stratum corneum can be detected in real time using a micro needle skin patch functionalized with an aptamer-coated carbon nanotube field effect transistor (CNT-FET) to be used for early diagnosis of various diseases It is an object of the invention.

국소적인 피부질환은 물론 다양한 질병상태에서 피부의 물질의 조성이 바뀌는 것이 보고되고 있다. 지질, 구조 단백질, 염증성 물질, 핵산, 대사산물 등 여러 물질이 질병상태에 따라 피부에서 다양하게 검출되는 것으로 알려져있다. 현재 아토피성 피부염, 흑색종, 피부의 세균성 염증 외에도 알츠하이머병, 파킨슨씨병, 유방암, 심혈관계 질환, 당뇨병, 약물 중독 등의 다양한 질환에서 피부의 바이오마커 분석이 이루어지고 있다.It has been reported that the composition of skin material changes not only in local skin diseases but also in various disease states. It is known that various substances such as lipids, structural proteins, inflammatory substances, nucleic acids, metabolites and the like are variously detected in the skin depending on the disease state. In addition to atopic dermatitis, melanoma and bacterial inflammation of the skin, biomarker analysis of skin has been performed in various diseases such as Alzheimer's disease, Parkinson's disease, breast cancer, cardiovascular disease, diabetes, drug addiction and the like.

본 발명은 피부의 각질층 (stratum corneum) 을 침투하여 표피층에 이를 수 있는 길이 약 200 μm의 Microneedle을 앱타머로 코팅된 탄소나노튜브 전계효과트랜지스터 (CNT-FET)로 기능화하여 피부내 세포간질에 존재하는 다양한 바이오마커가 이에 특이적으로 반응하는 앱타머와 결합했을 때 나타나는 전기전도도의 변화를 실시간으로 감지하여 이를 증폭시켜서 외부의 기기에 효과적으로 전송함으로써 다양한 질환을 조기에 진단할 수 있게 하는 진단용 스킨패치이다.The present invention relates to a method for manufacturing a carbon nanotube field effect transistor (CNT-FET) coated with an aptamer-coated carbon nanotube field effect transistor (CNT-FET) having a length of about 200 μm that penetrates the stratum corneum of the skin and reaches the skin layer. It is a diagnostic skin patch that detects the change of electrical conductivity when a variety of biomarkers are combined with an aptamer that reacts specifically with it, amplifies it, and effectively transmits it to an external device so that various diseases can be diagnosed early. .

본 발명을 통하여 알 수 있는 바와 같이, 본 발명의 패치는 피부내 세포간질에 존재하는 다양한 바이오마커가 이에 특이적으로 반응하는 앱타머와 결합했을 때 나타나는 전기전도도의 변화를 실시간으로 감지하여 이를 증폭시켜서 외부의 기기에 효과적으로 전송함으로써 다양한 질환을 조기에 진단할 수 있다.As can be seen from the present invention, the patch of the present invention senses the change in electrical conductivity when a variety of biomarkers existing in the intercellular epidermis bind to an aptamer specifically reacting thereto, So that various diseases can be diagnosed early.

도 1은 탄소나노튜브 표면에 링커를 부착시킨 후, 압타머를 이 링커에 고정하여 구성한 탄소나노튜브 압타머 전계트랜지스터. 이 경우 CDI-tween 20가 링커로 쓰였다.
도 2는 Microneedle 표면에 여러 종류의 압타머로 코팅된 탄소나노튜브 전계효과 트랜지스터를 부착한 후, 압타머가 특정 바이오마커와 결합했을 때 나타나는 전기전도도의 차이를 증폭해서 외부 기기 (스마트폰 등)에 전송해서 실시간으로 모니터링 하는 방법.
도 3은 Microneedle 내부의 채널에 여러 종류의 압타머로 코팅된 탄소나노튜브 전계효과 트랜지스터를 부착한 후, 압타머가 특정 바이오마커와 결합했을 때 나타나는 전기전도도의 차이를 증폭해서 외부 기기 (스마트폰 등)에 전송해서 실시간으로 모니터링 하는 방법.
BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a carbon nanotube extramammetic field effect transistor formed by attaching a linker to the surface of a carbon nanotube and fixing the extruder to the linker. In this case, CDI-tween 20 was used as a linker.
FIG. 2 is a graph showing the results obtained by attaching a carbon nanotube field effect transistor coated with various types of aptamers on a microneedle surface, amplifying a difference in electrical conductivity when an aptamer is combined with a specific biomarker, To monitor in real time.
FIG. 3 is a graph showing the relationship between the electric conductivity of a carbon nanotube field effect transistor coated with various types of aptamers on a channel inside a microneedle, To monitor in real time.

이하 비한정적인 실시예를 통하여 본 발명을 더욱 상세하게 설명한다.단 하기 실시예는 본 발명을 예시하기 위한 의도로 기재한 것으로서 본 발명의 범위는 하기 실시예에 의하여 제한되는 것으로 해석되지 않는다.
The present invention will now be described in more detail by way of non-limiting examples. The following examples are intended to illustrate the invention, but the scope of the invention is not to be construed as being limited by the following examples.

실시예Example 1.  One. 앱타머로Aptamer 코팅된 탄소나노튜브  Coated carbon nanotubes 전계효과트랜지스터Field effect transistor 제작 making

압타머를 분자인식물질로 이용한 탄소 나노튜브센서는 2005년 한국화학연구원에서 최초로 보고 하였다( Single-walled carbon nanotube biosensors using aptamers as molecular recognition elements. Journal of the American Chemical Society 127:11906-11907). 기판에 직접 성장된 탄소나노튜브에 포토리소그라피를 이용하여 전극과 절연층을 형성하였고, 탄소나노튜브의 표면에 표적물질을 검출하기 위하여 CDI-Tween 20을 링커로 사용하여 코팅한 후 압타머를 링커에 결합하였다 (그림 1).A carbon nanotube sensor using aptamer as a molecular recognition material was first reported by the Korean Chemical Research Institute in 2005 (Journal of the American Chemical Society 127: 11906-11907). An electrode and an insulating layer were formed on the carbon nanotube grown directly on the substrate. Photolithography was used to form an electrode and an insulating layer. To detect a target substance on the surface of the carbon nanotube, CDI-Tween 20 was coated as a linker, (Figure 1).

앱타머란 single strand DNA나 RNA의 삼차원구조를 이용해 특정 물질을 검출하는 방법으로 항원-항체 반응과 비슷하나 항체에 (10-15 nm) 비해 물질의 사이즈가 훨씬 작아서 (1-2 nm) 탄소나노튜브에 결합시켰을 때 훨씬 더 효과적으로 전계효과트랜지스터를 구현할 수 있는 장점이 있다.Aptamer is a method of detecting a specific substance by using a three-dimensional structure of single strand DNA or RNA. It is similar to an antigen-antibody reaction, but its size is much smaller than that of antibody (10-15 nm) It has the advantage of being able to implement a field effect transistor much more effectively when coupled to a tube.

또한 여러 종류의 바이오 마커에 대한 여러 종류의 앱타머를 코팅한 개개의 탄소나노튜브를 마이크로니들에 부착하여 (유전자칩과 비슷한 개념) 동시에 여러 종류의 물질을 검출할 수 있다 (Multiplexing). In addition, various types of substances can be detected simultaneously by attaching individual carbon nanotubes coated with various kinds of aptamers to various types of biomarkers (similar to a gene chip) by attaching them to a micro needle (Multiplexing).

실시예Example 2.  2. MicroneedleMicroneedle tip 표면에  tip on the surface 앱타머로Aptamer 코팅된 탄소나노튜브를 부착 Attach coated carbon nanotubes

각각의 마이크로니들 표면에 상기 압타머로 코팅된 탄소나노튜브를 부착시켰다.The carbon nanotubes coated with the aptamer were attached to the surface of each microneedle.

실시예Example 3.  3. MicroneedleMicroneedle 내부의 채널에  On the internal channel 앱타머로Aptamer 코팅된 탄소나노튜브를 부착 Attach coated carbon nanotubes

각각의 마이크로니들 내부의 채널에 압타머로 코팅된 탄소나노튜브를 부착시켰다.
Carbon nanotubes coated with an aptamer were attached to the channels inside each micro needle.

Claims (3)

마이크로니들(Microneedle)을 앱타머로 코팅된 탄소나노튜브 전계효과트랜지스터 (CNT-FET)로 기능화하여 피부 내 세포간질에 존재하는 바이오마커가 특이적으로 반응하는 앱타머와 결합했을 때 나타나는 전기전도도의 변화를 실시간으로 감지하여 이를 증폭시켜서 외부 기기에 효과적으로 전송함으로써 질환을 조기에 진단할 수 있게 하는 진단용 스킨패치.The change in electrical conductivity when a microneedle is functionalized with an aptamer-coated carbon nanotube field effect transistor (CNT-FET) and bound to an aptamer that specifically responds to a biomarker present in the intercellular epidermis of the skin In real time, amplifies it, and sends it to an external device effectively to diagnose the disease early. 제 1항에 있어서, 상기 바이오마커는 단백질, 펩타이드, DNA 또는 RNA인 것을 특징으로 하는 진단용 스킨패치.The diagnostic skin patch according to claim 1, wherein the biomarker is a protein, a peptide, DNA, or RNA. 제 1항에 있어서, 상기 외부 기기는 스마트폰, 휴대폰, 또는 무선통신기기인 것을 특징으로 하는 진단용 스킨패치.
The diagnostic skin patch according to claim 1, wherein the external device is a smart phone, a mobile phone, or a wireless communication device.
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WO2020069567A1 (en) 2018-10-02 2020-04-09 WearOptimo Pty Ltd Electrode arrangement
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