Patent Application
20210106969
The disclosure relates to a demethylation apparatus, a cancer therapy apparatus including the same, and a demethylation method and a cancer therapy apparatus and method using the same.
The terahertz electromagnetic wave (also referred to as terahertz wave hereinafter) has a wavelength between those of infrared ray and microwave, that is, a frequency ranging from 0.1 to 10 THz. Since the energy of terahertz wave is lower than the ionization level of biological sample, terahertz wave is used in biological system test. As a result, damage to biological samples caused by biological system test can be minimized.
The function of DNA may be controlled by a chemical change called methylation. When the excessive methylation phenomenon, in which the methylation level deviated from normal level, occurs, the function of DNA becomes abnormal and various diseases may occur.
It is an object of the invention to provide an apparatus and method capable of demethylating methylated molecules, and a cancer therapy apparatus and cancer therapy method using the same.
According to one aspect of the present invention, there is provided a demethylation method, comprising: (a) irradiating a terahertz wave of a first frequency band including a methylation resonance frequency onto a methylated molecule; and (b) demethylating the methylated molecule by the terahertz wave.
The demethylation method further comprises: (c) generating the terahertz wave using a laser beam; and (d) condensing and steering the terahertz wave by a polarizer toward the methylated molecule to be irradiated in (a).
The demethylation method further comprises: (e) filtering the terahertz wave condensed and steered in (d) to pass the terahertz wave of the first frequency band.
The first frequency band comprises a frequency of 1.6 THz.
According to another aspect of the present invention, there is provided a cancer therapy method comprising a demethylation method.
According to yet another aspect of the present invention, there is provided a cancer therapy apparatus capable of performing the demethylation method.
According to yet another aspect of the present invention, there is provided a demethylation apparatus irradiating a terahertz wave of a first frequency band including a methylation resonance frequency onto a methylated molecule to demethylate the methylated molecule by the terahertz wave.
The demethylation apparatus comprises: a laser beam generator generating a laser beam; a crystal excited by the laser beam to generate the a terahertz wave; a polarizer condensing and steering the terahertz wave generated by the crystal toward the methylated molecule to be irradiated onto the methylated molecule.
The demethylation apparatus further comprises a filter passing the terahertz wave of the first frequency band.
The filter is positioned in an optical path between the polarizer and the methylated molecule
The demethylation apparatus further comprises: a first half wavelength plate changing a polarization direction of the laser beam generated by the laser beam generator by 90 degrees; and a grating 300 tilting the laser beam having the polarization direction thereof changed by 90 degrees toward the crystal to be incident thereon.
The demethylation apparatus further comprises: an optical condenser disposed in an optical path between the grating and the crystal and condensing the laser beam from the grating to be incident on the crystal.
The demethylation apparatus further comprises: a second half wavelength plate disposed in an optical path between the optical condenser and the crystal and changing a polarization direction of the laser beam condensed by the optical condenser to a vertical polarization.
The terahertz wave comprises terahertz wave pulses or a continuous terahertz wave.
The first frequency band comprises a frequency of 1.6 THz.
According to yet another aspect of the present invention, there is provided a cancer therapy apparatus comprising the demethylation apparatus.
A demethylation apparatus capable of demethylating methylated molecules, a cancer therapy apparatus including the same, and a demethylation method and a cancer therapy method using the same are provided through the embodiments.
Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings such that those skilled in the art can easily implement the present invention. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms.
In order to clearly describe the present invention, parts unrelated to the description have been omitted, and like reference numerals indicates like parts throughout the specification. However, the present invention is not necessarily limited to the drawings.
The demethylation apparatus using a terahertz wave according to an embodiment irradiates a high-power terahertz electromagnetic wave onto methylated molecules, for example, methylated DNA for demethylation. Hereinafter, the high-power terahertz electromagnetic wave is referred to as a high-power terahertz wave. In the present disclosure, a methylation resonance frequency in a terahertz band is detected, and a demethylation reaction is induced by the high-power terahertz wave of a band including the corresponding resonance frequency.
First, a demethylation apparatus using a terahertz wave will be described with reference to
A high-power laser generated by a regenerative amplifier is irradiated onto a LiNbO3 crystal to generate a high-power terahertz wave in the demethylation apparatus 1. A power of the high-power terahertz wave may be measured by a pyroelectric detector in the demethylation apparatus 1. The demethylation apparatus 1 may induce a demethylation reaction by irradiating the high-power terahertz wave onto a methylated molecule (referred to as a sample hereinafter). In particular, since a terahertz wave of a certain band may be advantageous in inducing the demethylation reaction, a terahertz electromagnetic wave of the corresponding band may be generated by using a filter according to the embodiment.
As shown in
The laser beam generator 10 generates a high-power laser beam using a titanium:sapphire regenerative amplifier.
The half wavelength plate 100 changes the polarization direction of the high-power laser beam generated by the laser beam generator 10 by 90 degrees such that the laser beam has horizontal polarization before the laser beam is incident on the grating 300. The half wavelength plate 150 changes the polarization direction of the laser beam incident on the crystal 500 by 90 degrees such that the laser beam has a vertical polarization.
The optical chopper 200 intermittently passes i.e. intermittently blocks the laser beam passed through the half wavelength plate 100 at a predetermined period. As a result, the presence and the absence of the laser beam alternately occurs such that a lock-in amplifier (not shown) may generate a detection signal which has the same phase as a reference signal by rectifying the laser beam which has passed through the optical chopper 200.
According to one embodiment, the demethylation apparatus 1 may generate a terahertz wave pulse or a continuous terahertz wave. According to one embodiment, the demethylation method may relate to a method capable of showing that demethylation may be induced through a detection device using an arbitrary terahertz wave generator. The laser beam is pulsed using the optical chopper 200 to generate a high-power terahertz wave pulse. However, the present invention is not limited thereto, and the demethylation apparatus 1 may generate a continuous terahertz wave. In such case, the energy of a continuous terahertz wave may have a level at which demethylation of a sample is induced.
The grating 300 tilts to a specific angle the beam having horizontal polarization passed through the half wavelength plate 100. In order to generate the high-power terahertz wave using the crystal 500, the laser beam should be incident on the crystal 500 at a specific angle. For example, the grating 300 may have a grating angle and a grating spacing such that the laser beam is incident at an angle of 65 degrees with respect to the surface of the crystal 500.
The optical condenser 400 is located in an optical path between the grating 300 and the crystal 500 and narrows the laser beam from the grating 300. Since the laser beam passing through the grating 300 is too wide to be incident on the crystal 500, the optical condenser 400 uses a lens and a mirror to condense the beam to have a cross-section suitable to be incident on the crystal 500.
As shown in
The half wavelength plate 150 is located in the optical path between the optical condenser 400 and the crystal 500, and changes the polarization direction of the laser beam condensed by the optical condenser 400. As a result, the polarization direction of the laser beam incident on the crystal 500 passed through the half wavelength plate 150 is vertical.
The crystal 500 is excited by the incident laser beam and emits a terahertz wave. In the embodiment, crystal is used as an example of the terahertz wave generator, but the present invention is not limited thereto. The demethylation apparatus 1 may include various generators capable of generating high-power terahertz waves. For example, the generator may be a system that generates terahertz wave pulses using a crystal as in the embodiment, a system that generates continuous terahertz waves using a crystal or an air breakdown system. In addition, while a laser beam is used to generate a terahertz wave in the embodiment, the present invention is not limited thereto such that other means capable of generating a terahertz wave of a frequency band for inducing demethylation may be used.
The polarizer 600 may steer the terahertz wave radiated from the crystal 500 toward a predetermined direction using a reflective optical system. Here, the predetermined direction refers to an appropriate direction for irradiating the terahertz wave onto the sample 2.
The polarizer 600 shown in
The intensity of the terahertz wave radiated from the crystal 500 may be adjusted by two polarizing plates 601 and 602. The terahertz wave passing through the polarizing plates 601 and 602 is steered and condensed by the parabolic mirror 603, the reflector 604 and the parabolic mirror 605 and then irradiated onto the sample 2.
The pyroelectric detector 700 measures the terahertz wave that has passed through the sample 2 with an optical sensor. Information about the measured terahertz wave is fed back to the laser beam generator 10, and the laser beam outputted by the laser beam generator 10 may be controlled according to the information.
When compared with the embodiment shown in
According to the embodiments described with reference to
Like reference numerals indicates like parts described in
As shown in
The description of the parts the same as those described with reference to
The laser beam pulse from the laser beam generator 10 is split into two beams by the beam splitter 20. One of the two beams is a pump beam that excites the crystal 500 to generate a terahertz wave, and the other of the two beams is a probe beam that is irradiated to the pulse detector 900 to detect terahertz wave pulses. The terahertz wave pulse generated by the crystal 500 is incident on the sample, and then reflected from and/or transmitted through the sample. The terahertz wave pulse is modified by the reflection and/or transmission. The modified terahertz wave pulse is detected by the pulse detector 900.
The pump beam generated by the beam splitter 20 is steered by the reflector 21, and passes through the half wavelength plate 100, optical chopper 200, grating 300 and optical condenser 400 to be incident upon the crystal 500.
The delay element 30 is necessary for sampling the form of the terahertz wave pulse on a time axis, and generates the probe beam by delaying the laser beam outputted from the laser beam generator 10 and changing the path thereof. The probe beam may be used to scan and sample the terahertz wave pulses passed through the sample 2.
Since the length of the optical path is varied by the delay element 30, the time it takes for the probe beam to reach the pulse detector 900 is also varied. The embodiment may be applied to time-domain spectroscopy (TDS) for measuring the intensity of terahertz wave pulses incident on the pulse detector 900 while varying time.
The delay element 30 includes two reflectors 31 and 32, and the positions of the two reflectors 31 and 32 may be changed along the first axis (direction of the arrow shown in
The parabolic mirrors 901 and 902 condense terahertz wave pulses that have passed through the sample 2 and the condensed terahertz wave pulses are incident on the pulse detector 900. The probe beam from the reflector 33 passes through the parabolic mirror 901, is reflected by the parabolic mirror 902, and is incident on the pulse detector 900.
The pulse detector 900 detects the terahertz wave pulse that has passed through the sample 2. The pulse detector 900 measures the intensity of the incident terahertz wave pulse according to the change over time. The pulse detector 900 scans and samples the terahertz wave pulse that has passed through the sample 2 using the probe beam to measure a change in the terahertz wave pulse. When the signal corresponding to the intensity of the terahertz wave pulse measured by the pulse detector 900 is subjected to Fourier transform, the amplitude and phase of the spectrum of the transmitted terahertz wave may be obtained.
The pulse detector 900 may sample the terahertz wave pulses using an electro-optic sampling method. The pulse detector 900 includes a nonlinear crystal (e.g., ZnTe) 903 and a balanced photodetector 904.
The terahertz wave pulses as well as the laser beam transmitted through the sample 2 are incident on the nonlinear crystal 903. When the laser beam, which is the probe beam, and the terahertz wave pulses are incident on the nonlinear crystal 903, a phenomenon in which the polarization of the laser beam may change according to the electric field of the terahertz wave pulse due to the Pockels effect.
The photo detector 904 may indirectly measure the terahertz wave pulses by detecting a change in polarization of the laser beam using a quarter wavelength plate, a beam splitting polarizer, and a balance photodiode.
The photodetector 904 may measure the change over time in the intensity of the terahertz wave pulses according to TDS, and detect the amplitude and phase of each frequency component by subjecting the measured intensity to Fourier transform.
A demethylation reaction is induced by irradiating the high-power terahertz wave on the sample using the demethylation apparatuses shown in
When the terahertz wave is incident on methylated molecules, the resonance peak of the spectrum of the terahertz wave in a predetermined frequency band is determined according to the methylation degree of the methylated molecules. For example, as the methylation degree of DNA increases, the resonance peak gets higher around 1.6 THz (between 1.5 and 1.7 THz) in the frequency spectrum of the terahertz wave. Therefore, the demethylation degree of the methylated sample demethylated by the demethylation apparatus may be obtained through the terahertz wave measuring device.
Hereinafter, the effects of the demethylation apparatus will be described through experiment examples.
In the graph shown in
As shown in
The M-293T is divided into two samples, and a high-power terahertz wave is irradiated onto one of the M-293T samples for 1 hour using the demethylation apparatus shown in
As a result, as shown in
As shown in
Deoxycytidine and S-adenosyl-L-methionine (SAM) were prepared as substrates, and DNMT (DNA methyltransferase; New England Biolab) was added to prepare 5-Methyl-cytidine with a methyl group bonded to the 5 position of the cytosine according to the manufacturer's manual (https://www.neb.com/products/m0230-human-dna-cytosine-5-methyltransferase-dnmt1#Protocols %20&%20Manuals). This methylation process of Deoxycytidine is represented by the reaction scheme shown in
The intensity of the terahertz wave may be rapidly reduced by the moisture in the sample. In the experiment, a sample holder was used along with the terahertz wave measuring device of the demethylation apparatus. As shown in
For example, as shown in
The sample placed between the hydrophobic surface 54 of the quartz window 51 and the Teflon window 55 is cooled to 20 degrees below zero by the thermoelectric cooler 53.
As shown in (a) of
According to the spectrum shown in
As shown in (b) of
The “THz result” bar graphs denoted as “THz result” represent the methylation degree of DNA of the three samples measured using the terahertz wave measuring device, and the bar graphs denoted as “ELISA-like reaction” represent the methylation degree of the DNA of the three samples measured by the Elisa-like reaction method (hereinafter, referred to as ELISA quantitative method).
As shown in
As shown in
As shown in
As shown in
It is to be demonstrated that, with reference to
In the experiment in which the graphs shown
The spectrum shown in
As shown in
The spectrum shown in
As shown in
As described above, the high-power terahertz wave when radiated onto a sample induces the demethylation reaction. Considering the decrease of the resonance peak even when the filter is applied, it can be seen that the high-power terahertz wave of the predetermined frequency band (approximately 1.6 THz) is deeply involved in inducing demethylation.
The reason the demethylation degrees of the samples irradiated with terahertz wave using filter as shown in
14 to 18 are diagrams showing a graph showing a resonance frequency of a blood cancer sample and a resonance frequency of a demethylated sample, and a graph showing a comparison and verification of the methylation degree using a terahertz wave measuring device and ELISA quantification method.
As shown in
“SU-DHL1” in
In the bar graphs of
In
As shown in
The above-described demethylation apparatus and the demethylation method may be applied to a cancer therapy method and apparatus.
As shown in
The demethylation apparatus 1′ may be employed in cancer therapy apparatus 2 in place of demethylation apparatus 1.
The terahertz wave lens 21 condenses the high-power terahertz wave outputted from the demethylation apparatus 1.
The high-power terahertz wave condensed by the terahertz wave lens 21 travels along the waveguide 22. a high-power terahertz wave traveled along the waveguide 22 is irradiated onto the affected area where cancer cells are present via the terahertz wave output tube 23. The waveguide 22 may be extended to a predetermined length such that the terahertz wave output tube 23 may be positioned on the affected area. The user may hold and move the terahertz wave output tube 23 for treatment toward the affected area. The terahertz wave output tube 23 may be have a shape suitable for enabling such manipulation.
The embodiment shown in
Although the embodiment of the present invention has been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements by those skilled in the art using the basic concept of the present invention defined in the following claims are also in the scope of the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 10-2018-0040634 | Apr 2018 | KR | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/KR2018/014489 | 11/23/2018 | WO | 00 |
