Drug evaluation method and drug evaluation system for simulating change in blood concentration

Abstract

A drug evaluation method according to various embodiments of the present invention may comprise the steps of: receiving a pharmacokinetic (PK) profile, which is a blood concentration of a target drug over time; determining a compartment model from the PK profile; determining a parameter from the determined compartment model; and calculating, from the parameter, a liquid volume (Vr,1min) to be replaced every minute in a microfluidic system. A drug evaluation system according to various embodiments of the present invention comprises: an inlet for liquid injection into the system; an outlet for liquid discharge; a liquid reservoir for liquid storage; a magnetic stirrer arranged in the reservoir; a pump for controlling liquid flow; and a control unit for controlling the speed of the pump, wherein the control unit can perform the operation of the above-described drug evaluation method.

Description

Drug evaluation method and drug evaluation system for simulating changes in blood concentration

[0001] The present invention relates to a drug evaluation method and a drug evaluation system for simulating changes in blood concentration.

[0002] Traditional in vitro experimental methodologies have served as crucial tools in the drug development process. These methods primarily involve assessing the biological effects and toxicity of drugs through in vitro cell cultures. However, these methods have several limitations.

[0003] First, most in vitro experiments evaluate drug effects at a single concentration. This does not accurately reflect the actual distribution of drugs in vivo, nor does it account for changes in drug dose or concentration over time. In a real biological environment, drug concentration continuously changes after injection, and these changes significantly impact drug effects.

[0004] Second, most existing in vitro systems evaluate drug effects in static environments. This fails to reflect dynamic factors, such as blood circulation in a living body. Specifically, drugs injected into the blood undergo rapid concentration changes in vivo. Current cell culture systems operate at a single concentration, failing to reflect these rapid changes and thus falling short of the true in vivo changes.

[0005] Thus, existing in vitro experimental methodologies have limitations in accurately reflecting the efficacy and toxicity of drugs in real-world biological environments. This represents a critical issue in drug development, highlighting the need for new experimental methodologies to address this issue.

[0006] Meanwhile, animal testing is an essential element in the drug development process, playing a crucial role in verifying efficacy and safety. However, animal testing poses numerous ethical challenges.

[0007] First, animal testing can cause suffering to the animals. Animals experience discomfort and pain during the experiment, and drug administration experiments, in particular, can cause serious side effects. The ethical concerns surrounding this issue are a frequent topic of discussion among scientists.

[0008] Furthermore, animal testing often results in the unnecessary sacrifice of large numbers of animals. At every stage of the drug development process, numerous animals are used in experiments, and this massive sacrifice fundamentally undermines the dignity of animals for human-centered interests.

[0009] Thus, animal testing raises complex ethical questions about balancing the needs of drug development with animal welfare. To address this, the development of new experimental methodologies that can replace animals is needed.

[0010] Meanwhile, drug delivery methods continue to evolve over time, and these changes highlight the need for new experimental methodologies. While drugs were previously administered primarily as solutions dissolved in liquid, recent research is actively underway to formulate drugs into nanoparticles.

[0011] Nanoparticle drug formulations hold great potential for maximizing drug efficacy and minimizing side effects. These formulations allow for more effective control over drug consumption and distribution, playing a crucial role in the treatment of diseases such as cancer. However, the PK profiles of these nanoparticle formulations differ significantly from those of conventional solution-type drugs, raising the need for new experimental methodologies.

[0012] Additionally, one of the unique properties of nanoparticle drugs is the enhanced permeability and retention (EPR) effect, which allows nanoparticles to better penetrate and remain in tumor tissue. While this offers significant advantages in cancer treatment, new experimental methods are required to accurately assess this effect.

[0013] These changes in drug administration methods and the resulting need for new experiments demonstrate the limitations of current experimental methodologies, highlighting the importance of developing new experimental methodologies that can replace animals.

[0014] The purpose of the present invention is to solve the above-described problems, and to provide a drug evaluation method and drug evaluation system that can overcome the limitations and problems of in vitro experiments, resolve ethical issues of animal experiments, and satisfy new drug evaluation methods according to changes in drug administration methods.

[0015] A drug evaluation method according to various embodiments of the present invention comprises the steps of: receiving a pharmacokinetic (PK) profile, which is a blood concentration of a target drug over time; determining a compartment model from the PK profile; determining parameters from the determined compartment model; and determining a liquid volume (V) to be replaced every minute in a microfluidic system from the parameters. r,1min ) may include a step of calculating.

[0016] A drug evaluation system according to various embodiments of the present invention includes an inlet for injecting liquid into the system; an outlet for discharging liquid; a liquid reservoir for storing liquid; a magnetic stirrer disposed within the reservoir; a pump for controlling the flow of liquid; and a control unit for controlling the speed of the pump, wherein the control unit can perform the operations of the drug evaluation method described above.

[0017] The drug evaluation method and system of the present invention are microfluidic systems that actually reflect PK profiles, replacing animal testing and providing more realistic drug responses. This system can more accurately assess drug responses across various drug formulations. Furthermore, because the drug evaluation system of the present invention reflects PK profiles across drug formulations, it provides more reliable data and improves experimental efficiency and accuracy.

[0018] The drug evaluation method and system of the present invention can reduce animal testing while simultaneously improving experimental accuracy and efficiency. Furthermore, it can improve the drug development process and more accurately assess drug safety and efficacy.

[0019] In addition, by equipping the drug evaluation method and drug evaluation system of the present invention with an additional configuration capable of measuring the EPR effect, the cancer treatment effect of the nanoparticle drug can be evaluated more accurately.

[0020] Figure 1 is a schematic diagram of a microfluidic system to which the drug evaluation method of the present invention is applied.

[0021] Figure 2 is a flowchart of the operation of a drug evaluation method according to various embodiments of the present invention.

[0022] Figure 3 is a drawing for explaining the compartment model.

[0023] Figure 4 is a drawing for explaining a two-compartment model.

[0024] Figure 5 is a graph for explaining the drug evaluation method of the present invention in the case of a two-compartment model.

[0025] Figure 6 compares the results of using the drug evaluation system of the present invention with the results of animal experiments.

[0026] Hereinafter, various embodiments of this document are described with reference to the attached drawings. The embodiments and terminology used herein are not intended to limit the technology described in this document to specific embodiments, but should be understood to encompass various modifications, equivalents, and / or alternatives of the embodiments.

[0027] Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings.

[0028]

[0029] The drug evaluation method and system according to various embodiments of the present invention are methods and systems capable of reflecting the pharmacokinetic (PK) profile of a drug in a microfluidic system. Specifically, the drug evaluation method and system of the present invention can accurately assess the efficacy and safety of a drug by monitoring and modeling changes in blood drug concentration in vitro in real time.

[0030] Referring to Figure 1, a microfluidic system to which the drug evaluation method of the present invention is applied is described. A microfluidic system is a device capable of controlling and manipulating liquids using small channels. Microfluidic systems offer high precision and fine fluid flow control, simplifying complex experimental procedures and enabling the efficient processing of extremely small samples. Microfluidic systems utilize millimeter-sized tubes to transport liquids.

[0031] Referring to Figure 1, the microfluidic system may be composed of an inlet for injecting liquid into the system, an outlet for discharging liquid, a liquid reservoir for storing excess liquid outside the tube, a magnetic stirrer for quickly and uniformly mixing a drug with the liquid, and a multi-channel pump for providing various flows within the microfluid.

[0032] Microfluidic systems can be equipped with various organ-on-a-chips or tumor-on-a-chips. These chips are devices designed to mimic the functions of specific tissues or cells. If microfluidic systems are like artificial blood vessels, then the chips can be like artificial organs. Each of these on-a-chips has a specific purpose, and they can be used to simultaneously verify the anticancer effects and toxicity of drugs. The examples are not limited to this, and microfluidic systems can also be equipped with electronic chips. This can add the ability to monitor drug concentrations within the system in real time.

[0033] By applying the drug evaluation method of the present invention to a microfluidic system, a liquid flow simulating blood flow in the body can be implemented, thereby reflecting the actual body environment.

[0034]

[0035] Referring to FIG. 2, a drug evaluation method according to various embodiments of the present invention comprises the steps of: receiving a pharmacokinetic (PK) profile, which is a blood concentration of a target drug over time (S100); determining a compartment model from the PK profile (S200); determining parameters from the determined compartment model (S300); and determining a liquid volume (V) to be replaced every minute in a microfluidic system from the parameters. r,1min ) may include a step (S400) of calculating.

[0036] First, in the step of receiving a PK profile (S100), blood concentration profile data measured over time by administering the drug to an animal can be received. A PK profile is data that describes changes in blood concentration after drug administration. Since blood drug concentration is directly related to the drug's efficacy, it plays a crucial role in accurately understanding and predicting drug efficacy.

[0037] Next, in the step of determining the compartment model (S200), the compartment model can be determined based on the graph shape of the preceding PK profile. Specifically, referring to FIG. 3, the compartment model can be divided into a 1-compartment model and a 2-compartment model. The 1-compartment model is a model in which the natural logarithm of the blood concentration over time is expressed as a straight line or close to a straight line. The 2-compartment model is a model in which the natural logarithm of the blood concentration over time is not expressed as a straight line. In other words, the 2-compartment model is a curved or broken graph model.

[0038] Next, in the step of determining the parameters (S300), the amount of drug to be administered into the microfluidic system (D system ) and the drug elimination rate constant k can be determined.

[0039] Specifically, in the step of determining parameters (S300), the parameters can be determined in different ways depending on the type of compartment model below.

[0040] First, let's explain the 1-compartment model.

[0041] For drugs that follow the 1-compartment model, the blood concentration (C) of the drug at a specific time is given by the following equation: t ) decreases exponentially over time.

[0042] C t = C 0 × e -kt

[0043] Here, C0 is the initial concentration, k is the rate constant, t is time, and e is the natural constant (e=2.7182..).

[0044] That is, in the case of a 1-compartment model, the drug elimination rate constant k can be determined using the following equation in the step of determining parameters (S300).

[0045] k= {ln(C0) - ln(C t )} / t

[0046] (Here, C0 is the initial concentration, which is the blood concentration at time 0, C t is the blood concentration at a specific time (t), where t is time.)

[0047] Meanwhile, as can be seen from the above equation, when a drug is injected intravenously into an animal, it is immediately distributed to the blood and various organs of the body through rapid blood flow, thereby forming a superconcentration (C0), which is the drug concentration in the blood at time 0. In pharmacokinetics, the amount of drug administered to an animal (D animal ) and C0, the following equation holds.

[0048] D animal = Vd × C0

[0049] Here, Vd is a pharmacokinetic term with a volume unit representing the volume of distribution, which refers to the relationship between the administered dose and the initial concentration in the body. A larger value indicates that a greater amount of the drug has been distributed to organs other than the blood at time 0. Therefore, the volume may exceed the actual body fluid volume of the animal.

[0050] Drugs injected into a microfluidic system simulating the body should be immediately and uniformly distributed throughout the entire system to form a super-concentration (C0) similar to the values ​​obtained in animals. However, unlike the body environment, there are no various organs in the microfluidic system other than the fluid to which the drug is distributed, so the actual total volume of fluid in the system (V) rather than the Vd value system) must be applied to obtain the same C0 value. In addition, to immediately achieve concentration equilibrium within the entire fluid, the drug is injected into a reservoir containing a large volume of liquid within the microfluidic system. A magnetic stirrer is installed within the reservoir so that the drug can be quickly and uniformly mixed with the liquid. The amount of drug (D) to be administered within the microfluidic system system ) is calculated by the following formula.

[0051] D system = V system × C0

[0052] In the 1-compartment model, the concentration decreasing over time is always a constant ratio of the initial concentration. As can be seen from the equation, the drug concentration C after time t is equal to the initial concentration C0 multiplied by e^(-kt). In other words, the decrease in drug concentration occurs through first-order loss, and this loss is implemented in the microfluidic system by removing a constant volume of liquid every minute and adding a new liquid that does not contain the same amount of drug. At this time, the total liquid volume and the k value are used to calculate the volume of liquid that must be replaced per minute. The unit of the k value, which is a function of time, is min. -1 is calculated by converting it into V . That is, for the 1-compartment model, the liquid volume (V ) that must be replaced per minute within the microfluidic system r,1min ) can be calculated using the following formula in the step (S400).

[0053] V r,1min = V system × (1 - e -k )

[0054] Through this, the volume of liquid that must be replaced per minute (Volume of replacement per 1 min, V r,1min) can be obtained. In the 1-compartment model, since the drug concentration decreases at the same rate from beginning to end, the volume of liquid discharged to the outlet and the volume of new liquid flowing into the inlet for 1 minute are V r,1min This can be maintained without significant change. Through this, the microfluidic system can accurately reflect the drug PK profile obtained in animals by exponentially decreasing the drug's blood concentration over time.

[0055]

[0056] Below, the two-compartment model is described.

[0057] In general, the two-compartment model is used when the natural logarithm of the drug concentration in the blood after the drug is injected is not linear, or when the drug concentration in the blood is fitted by an exponential function (i.e., C t = C0*?* e -kt ) cannot be expressed as a linear model. That is, when the natural logarithm of the blood drug concentration over time is not linear, a multi-compartment model is used to represent the concentration changing over time. This model mainly consists of a 'central compartment' where the drug is well distributed and a 'peripheral compartment' where the drug is less distributed. The central compartment includes tissues where the drug is distributed quickly and actively, and the peripheral compartment refers to tissues where the drug is distributed relatively slowly and poorly, such as muscle or adipose tissue.

[0058] Referring to Figure 4, when the blood concentration (Ct) of a drug following the two-compartment model at a specific time is expressed on the log scale of the y-axis, there is a part where there is a single bend in the middle. The section before this point is expressed as the pre-phase of distribution (α phase), and the section after it is expressed as the post-phase of distribution (β phase). Immediately after a drug following the two-compartment model is administered into the body, at time 0, drug distribution to the central compartment organs occurs immediately, forming a superconcentration (C0). In the pre-phase of distribution, drug transport from the central compartment to the peripheral compartment occurs gradually, and in the post-phase of distribution, after drug transport to the peripheral compartment is complete, only overall drug loss occurs.

[0059] A two-compartment curve can be expressed as the sum of two first-order vanishing graphs by applying the residual method, as shown in Figure 4. By extending the straight line portion of the distribution posterior as shown in the blue line, the slope β and y-intercept B can be obtained, and the residual line obtained by subtracting the blue line from the purple line is shown in red to obtain its slope α and y-intercept A.

[0060] Red line (1st graph) Ae -αt , the blue line (second graph) is Be -βt It can be quantified as , and since the 2-compartment curve is the sum of two straight lines, for drugs that follow the 2-compartment model, it can be expressed as follows.

[0061] C t = Ae -αt + Be -βt

[0062] To implement concentration changes in a two-compartment microfluidic system, the superconcentration (C0) must be determined, as in the one-compartment. The superconcentration in the two-compartment can be determined in several ways, as follows.

[0063] C0= A + B

[0064] D animal = Vd × C0

[0065] As with the 1-compartment model, unlike the body environment, there are no various organs in the microfluidic system other than the fluid to which the drug is distributed, so the actual total volume of fluid in the system (V ) rather than the Vd value is used. system ) must be applied to obtain the same C0 value. Therefore, the amount of drug (Dsystem) that must be injected into the system can be calculated as follows.

[0066] D system = V system × C0

[0067]

[0068] Meanwhile, in the case of a two-compartment model with two exponential terms, the decrease in drug concentration in the blood is more complex and requires a pump speed that varies over time. To achieve this, the pump speed can be adjusted for each interval between blood sampling times in animal experiments. This allows for varying the fluid volume to be replaced per hour for each interval.

[0069] For example, referring to Fig. 5, C0, C1, C2, C3, C4, and C5 represent drug concentrations at each sampling point. That is, C0 is the initial concentration, C1 is the blood concentration at t1, C2 is the blood concentration at t2, C3 is the blood concentration at t3, C4 is the blood concentration at t4, and C5 is the blood concentration at t5. k1, k2, k3, k4, and k5 represent the drug elimination rate constants (min) in the first interval from time 0 to t1, the second interval from t1 to t2, the third interval from t2 to t3, the fourth interval from t3 to t4, and the fifth interval from t4 to t5, respectively. -1 ) is indicated.

[0070] At this time, the disappearance rate constant in each section can be obtained as follows. That is, in the case of a two-compartment model, the disappearance rate constant k of the drug can be determined using the following formula in the step of determining parameters (S300).

[0071] k 1= {ln(C0)-ln(C1)} / (t1-0)

[0072] k 2= {ln(C1)-ln(C2)} / (t2-t1)

[0073] k 3= {ln(C0)-ln(C1)} / (t3-t2)

[0074] k 4= {ln(C1)-ln(C2)} / (t4-t3)

[0075] k 5= {ln(C1)-ln(C2)} / (t5-t4)

[0076] Next, for the two-compartment model, the liquid volume (V) that must be replaced per minute within the microfluidic system r,1min ) In the step (S400) of calculating, the calculation can be performed for each of the first to fifth sections using the following formula.

[0077] V in the first section r,1min = V system × (1 - e -k1 )

[0078] V in the second section r,1min = V system × (1 - e -k2 )

[0079] V of the third section r,1min = V system × (1 - e -k3 )

[0080] V of the 4th section r,1min = V system × (1 - e -k4 )

[0081] V of the 5th section r,1min = V system × (1 - e -k5 )

[0082] In this way, V in each section r,1minBy controlling the blood concentration of a drug within a microfluidic system, changes in the blood concentration can be implemented very similarly to reality. Meanwhile, although the number of sampling points is described as 5 in the above, the embodiment is not limited thereto, and the number of sampling points may be 2 or more. Preferably, the number of sampling points may be 5 to 10.

[0083] Meanwhile, in the previous example, the time interval was divided and the liquid volume (V) that must be replaced every minute for each interval r,1min ) was calculated, but according to another embodiment, V changes over time r,1min By calculating the values, we can also implement more subtle concentration changes.

[0084] Specifically, the time (t) after administration n minutes as follows: n ) in V r,1min can induce.

[0085]

[0086] Time t from the 2-compartment model equation n , t (n+1) Concentration C at time n and C( n+1) Each can be obtained as follows.

[0087] C n = Ae -αtn + Be -βtn

[0088] C (n+1) = Ae -αt(n+1) + Be -βt(n+1)

[0089] Also, the decay rate constant (k) at time n minutes n ) can be obtained as follows.

[0090] k n =ln(C n )-ln(C (n+1) )

[0091] =ln(C n / C (n+1) )

[0092] Cn , C (n+1) If we substitute it into the above equation, we get the following.

[0093] k n =ln{(Ae -αtn +Be -βtn ) / (Ae -αt(n+1) +Be -βt(n+1) )}

[0094] (Here, k n is the decay rate constant at time n minutes, α is the slope of the first graph to which the residual method is applied, A is the y-intercept of the first graph, β is the slope of the second graph to which the residual method is applied, B is the y-intercept of the second graph, tn is time n, and tn+1 is time n+1.)

[0095] k above n Using the value V at time n r,1min can be obtained.

[0096] V at time n r,1min = V system × (1 - e -kn )

[0097] Using the equation above, the k value and V that change every minute r,1min By reflecting this, more sophisticated concentration changes can be implemented.

[0098]

[0099] A drug evaluation system according to various embodiments of the present invention is characterized in that the above-described microfluidic system is driven by the above-described drug evaluation method. That is, the drug evaluation system according to various embodiments of the present invention includes an inlet for injecting liquid into the system; an outlet for discharging liquid; a liquid reservoir for storing liquid; a magnetic stirrer disposed within the reservoir; a pump for controlling the flow of liquid; and a control unit for controlling the speed of the pump, wherein the control unit can perform the operation of the above-described drug evaluation method.

[0100] Specifically, the control unit can first receive a PK profile. The control unit can receive blood concentration profile data measured over time by administering a drug to an animal.

[0101] The control unit can determine a compartment model from the PK profile. The control unit can determine, from the PK profile, either a 1-compartment model in which the natural logarithm of the blood concentration over time is expressed as a straight line; or a 2-compartment model in which the natural logarithm of the blood concentration over time is not expressed as a straight line.

[0102] The control unit can determine parameters from the determined compartment model. The control unit can determine the amount of drug (D) to be administered into the microfluidic system from the determined compartment model. system ) and the drug elimination rate constant k can be determined.

[0103] Specifically, the control unit controls the amount of drug (D) to be administered into the microfluidic system. system ) can be determined in the following manner.

[0104] D system = V system × C0

[0105] (Here, V system is the total volume of actual fluid in the microfluidic system, and C0 is the hyperconcentration, which is the blood concentration at time 0.)

[0106] Meanwhile, if the control unit determines the PK profile as a 1-compartment model, the drug elimination rate constant k can be determined by the following equation.

[0107] k= {ln(C0) - ln(C t )} / t

[0108] (Here, C0 is the initial concentration, which is the blood concentration at time 0, C t is the blood concentration at a specific time (t), where t is time.)

[0109] The control unit calculates the volume of liquid (V) that must be replaced per minute in the microfluidic system from this dissipation rate constant k. r,1min ) can be determined in the following manner.

[0110] V r,1min = V system × (1 - e -k )

[0111] (Here, V system is the total volume of actual fluid in the microfluidic system, and k is the elimination rate constant of the drug.)

[0112] Meanwhile, when the control unit determines the PK profile as a two-compartment model, the dissipation rate constant k1 in the first section from time 0 to time t1 and the dissipation rate constant k2 in the second section from time t1 to time t2 can be determined by the following equations.

[0113] k 1= {ln(C0)-ln(C1)} / (t1-0)

[0114] k 2= {ln(C1)-ln(C2)} / (t2-t1)

[0115] (Here, C0 is the initial concentration, C1 is the blood concentration at t1, and C2 is the blood concentration at t2.)

[0116] The control unit is configured to have these dissipation rate constants k1 and The volume of liquid that must be replaced per minute in the microfluidic system from k2 (V r,1min ) can be determined in the following manner.

[0117] V in the first section r,1min = V system × (1 - e -k1 )

[0118] V in the second section r,1min = V system × (1 - e -k2 )

[0119] (Here, V systemis the total volume of the actual fluid in the microfluidic system, k1 is the disappearance rate constant of the drug in the first section, and k2 is the disappearance rate constant of the drug in the second section.)

[0120] Alternatively, the control unit determines the drug elimination rate constant k from two graphs using the residual method when the PK profile is determined by a two-compartment model. n It can be decided in this way.

[0121] k n =ln{(Ae -αtn +Be -βtn ) / (Ae -αt(n+1) +Be -βt(n+1) )}

[0122] (Here, k n is the decay rate constant at time n minutes, α is the slope of the first graph to which the residual method is applied, A is the y-intercept of the first graph, β is the slope of the second graph to which the residual method is applied, B is the y-intercept of the second graph, tn is time n, and tn+1 is time n+1.)

[0123] The control unit is, these k n Using the value, V at time n according to the following equation r,1min can be obtained.

[0124] V r,1min = V system × (1 - e -kn )

[0125] The control unit calculates V r,1min The liquid volume can be controlled by controlling the pump, inlet and outlet, etc. using .

[0126] The drug evaluation method and system of the present invention, a microfluidic system that actually reflects PK profiles, can replace animal testing and provide more realistic drug responses. This system can more accurately assess drug responses across various drug formulations. Furthermore, because the drug evaluation system of the present invention reflects PK profiles across drug formulations, it can provide more reliable data and improve experimental efficiency and accuracy.

[0127] The drug evaluation method and system of the present invention can reduce animal testing while simultaneously improving experimental accuracy and efficiency. Furthermore, it can improve the drug development process and more accurately assess drug safety and efficacy.

[0128] In addition, by equipping the drug evaluation method and drug evaluation system of the present invention with an additional configuration capable of measuring the EPR (enhanced permeability and retention) effect, the cancer treatment effect of the nanoparticle drug can be evaluated more accurately.

[0129]

[0130] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0131]

[0132] Experimental method

[0133] 1. Establishment of HPLC analysis conditions for paclitaxel

[0134] HPLC (High Performance Liquid Chromatography) is the most versatile and widely used elution chromatography technique, boasting the advantages of high sensitivity, easy and accurate quantitation, and ease of automation. Therefore, this experiment aimed to establish analytical conditions for paclitaxel using HPLC. A commercially available, high-purity standard, paclitaxel, was used, and the analytical conditions were established through a process of stepwise dilution and confirming the appearance of a peak during HPLC analysis.

[0135] 2. Intravenous drug administration to ICG mice

[0136] After diluting the drug to an appropriate concentration, it was administered via the tail vein of ICG mice. A syringe was used to insert the needle into the vein, slowly injecting the drug, and ensuring no adverse reactions occurred. All procedures were performed under appropriate sterile conditions, and the injection site was disinfected to reduce the risk of infection. After the experiment, used needles, syringes, and other potentially contaminated materials were disposed of according to regulations.

[0137] 3. Calculation of PK parameters from blood concentration profile obtained after intravenous administration

[0138] Immediately after drug administration to ICG mice, blood samples were collected every 0, 5, 15, 30, 60, 120, and 240 minutes to measure blood concentrations at each hour. A concentration-time curve was drawn using the drug concentrations of the measured blood samples to show the distribution, metabolism, and elimination of the drug over time. From the blood concentration profile, the elimination rate constant (k) and volume of distribution (V) were calculated. d ) and removal rate (CL t ) and other pharmacokinetic parameters were calculated.

[0139] 4. Calculating the flow rate within the system by reflecting the PK parameter

[0140] PK parameters were calculated from drug concentrations in blood samples, and the hourly concentration decrease rate was calculated by reflecting the elimination rate constant (k). Based on the calculated hourly concentration decrease rate, the flow rate was assigned according to drug elimination at each point, and the results are presented in Table 1 below.

[0141] Interval (min)k(min) -1 )V r,1min (ml / min)0~50.1060092.0116660015~150.0965661.84099694615~300.0688581.33082469730~600.02555 10.50455637360~1200.0105490.209878546120~2400.0131950.262160923240~4800.0008550.017094503

[0142] 5. Media collection and concentration analysis after drug administration within the fluid flow system As with the PK parameters determined by intravenous drug administration to ICG mice, we attempted to mimic the PK parameters by administering drugs into the reservoir of the fluid flow system. The reservoir contained an amount of medium (RPMI1640) equivalent to the normal blood volume of the ICG mouse, and the drug was injected at the same concentration as that administered to the ICG mouse. Media was collected over time and the concentration was analyzed via HPLC.

[0143]

[0144] Experimental results

[0145] 1. HPLC analysis conditions

[0146] A standard calibration curve was prepared using the peak area by concentration obtained from the chromatogram of the standard solution, and the presence of paclitaxel was confirmed through this. A mobile phase of ACN:DW=70:30 was used, and the analyte was detected at a wavelength of 227.4 nm, which was established as the analysis condition.

[0147] 2. PK profile and parameter calculation after intravenous drug administration to ICG mice

[0148] After intravenous administration of the drug to ICG mice, blood samples were collected hourly to measure drug concentrations. The PK profile and PK parameter values ​​obtained through this are as follows.

[0149]

[0150] Here, A, B, α, and β were obtained by fitting the blood concentration-time actual values. Specifically, β was derived by obtaining the slope of the straight line from the second graph, which is a straight line connecting the data in the posterior distribution (β phase), and B was obtained by reading the y-intercept.

[0151] C β =B·e -β·t

[0152] Next, the first graph can be derived by plotting the data obtained by subtracting the extended linear data of the posterior distribution (β phase) from the actual blood concentration-time measured values, and can be expressed by the following equation.

[0153] CC β =CB·e -β·t =A·e -α·t

[0154] Cr=A·e -α·t

[0155] Through this, the first graph is derived, α is derived by multiplying the slope of the first graph by 2.303, and A is obtained by reading the y-intercept.

[0156] From A, B, α, and β, parameters such as half-life, distribution volume, and clearance in the α phase and β phase can be calculated.

[0157]

[0158]

[0159] Meanwhile, CL tis the clearance rate, which is the volume of blood purified per unit time (ml / min), and is defined as (drug elimination rate) / (drug blood concentration), and can be expressed by the following equation.

[0160]

[0161] If we integrate the above equation from t=0 to t=∞, we get the following.

[0162]

[0163] Here, D represents the dose and AUC represents the area under the blood concentration-time curve.

[0164] 3. Changes in concentration within the system after drug administration in the fluid flow system and comparison with animal data

[0165] After drug administration within the fluid flow system, samples were collected hourly to measure concentration. The PK mimic profile and parameter values ​​of the fluid flow system obtained through this process are as follows.

[0166]

[0167] The results of expressing this as a concentration-time curve and comparing them are as shown in Figure 6.

[0168] Referring to Figure 6, it can be confirmed that the error of the AUC value is 6.19%, and the error of the CL value is 5.83%, both values ​​being less than 10%. In other words, from a pharmacodynamic perspective, the experimental results of the fluid flow system are similar to the results of the animal experiment, and the error range is less than 10%, confirming that the fluid flow system can sufficiently replace animal experiments.

[0169]

[0170] The present invention has been described above, focusing on preferred embodiments thereof. Those skilled in the art will appreciate that the present invention can be implemented in modified forms without departing from its essential characteristics. Therefore, the disclosed embodiments should be considered illustrative rather than limiting. The scope of the present invention is set forth in the claims, not the foregoing description, and all differences within the scope equivalent thereto should be construed as being encompassed by the present invention.

[0171] The drug evaluation method and drug evaluation system of the present invention are microfluidic systems that actually reflect PK profiles, and thus can replace animal testing and more accurately evaluate drug responses according to various drug formulations, thus having high potential for industrial use.

Claims

1. A step of receiving a pharmacokinetic (PK) profile, which is a blood concentration profile of a target drug over time; A step of determining a compartment model from the above PK profile; A step of determining parameters from the determined compartment model; and From the above parameters, the liquid volume (V) that must be replaced per minute in the microfluidic system r,1min ) comprising the step of calculating a drug evaluation method.

2. In paragraph 1, The step of determining the above compartment model is: A 1-compartment model in which the natural logarithm of blood concentration over time is expressed as a straight line; and A drug evaluation method characterized in that the natural logarithm of blood concentration over time is determined by one of the two-compartment models that is not expressed by a straight line.

3. In paragraph 1, The step of determining the above parameters is: The amount of drug to be administered within the microfluidic system (D system ) and a drug evaluation method characterized by determining the drug elimination rate constant k.

4. In paragraph 3, The amount of drug to be administered into the above microfluidic system (D system ) is a drug evaluation method characterized by being determined by the following formula. D system = V system × C0 (Here, V system is the total volume of actual fluid in the microfluidic system, and C0 is the hyperconcentration, which is the blood concentration at time 0.) 5. In paragraph 2, The step of determining the above parameters is: A drug evaluation method characterized in that, for the above 1-compartment model, the drug elimination rate constant k is determined by the following equation. k= {ln(C0) - ln(C t )} / t (Here, C0 is the initial concentration, which is the blood concentration at time 0, C t is the blood concentration at a specific time (t), where t is time.) 6. In paragraph 2, The step of determining the above parameters is: For the above 2-compartment model, The vanishing velocity constant k1 in the first interval from time 0 to time t1; and A drug evaluation method characterized in that the disappearance rate constant k2 in the second section from time t1 to time t2 is determined by the following equation. k 1= {ln(C0)-ln(C1)} / (t1-0) k 2= {ln(C1)-ln(C2)} / (t2-t1) (Here, C0 is the initial concentration, C1 is the blood concentration at t1, and C2 is the blood concentration at t2.) 7. In paragraph 2, The volume of liquid (V) that must be replaced per minute within the above microfluidic system r,1min ) in the calculation step, For the above 1-compartment model, a drug evaluation method characterized by calculating using the following equation. V r,1min = V system × (1 - e -k ) (Here, V system is the total volume of actual fluid in the microfluidic system, and k is the elimination rate constant of the drug.) 8. In paragraph 2, The volume of liquid (V) that must be replaced per minute within the above microfluidic system r,1min ) in the calculation step, For the above 2-compartment model, V in the first interval from time 0 to time t1 r,1min ; and V in the second interval from time t1 to time t2 r,1min A drug evaluation method characterized in that each of the following equations is determined. V in the first section r,1min = V system × (1 - e -k1 ) V in the second section r,1min = V system × (1 - e -k2 ) (Here, V system is the total volume of the actual fluid in the microfluidic system, k1 is the disappearance rate constant of the drug in the first section, and k2 is the disappearance rate constant of the drug in the second section.) 9. In paragraph 2, The step of determining the above parameters is: For the above two-compartment model, the drug elimination rate constant k is obtained from two graphs using the residual method. n A drug evaluation method characterized by determining the following. k n =ln{(Ae -αtn +Be -βtn ) / (Ae -αt(n+1) +Be -βt(n+1) )} (Here, k n is the decay rate constant at time n minutes, α is the slope of the first graph to which the residual method is applied, A is the y-intercept of the first graph, β is the slope of the second graph to which the residual method is applied, B is the y-intercept of the second graph, tn is time n, and tn+1 is time n+1.) 10. In paragraph 9, The volume of liquid (V) that must be replaced per minute within the above microfluidic system r,1min ) in the calculation step, For the above 2-compartment model, The above k n Using the value, V at time n according to the following equation r,1min A drug evaluation method characterized by obtaining . V r,1min = V system × (1 - e -kn ) 11. Inlet for injecting liquid into the system; Outlet for liquid discharge; Liquid reservoir for liquid storage; A magnetic stirrer placed within the reservoir; a pump that regulates the flow of liquid; and A control unit for controlling the speed of the above pump is included, The above control unit, The volume of liquid that must be replaced per minute in a microfluidic system (V r,1min ) is characterized by performing an operation of calculating a drug evaluation system.

12. In paragraph 11, The above control unit, Receive the PK profile, Determine the compartment model from the above PK profile, Determine the parameters from the determined compartment model, From the above parameters, the liquid volume (V) that must be replaced per minute in the microfluidic system r,1min ) is characterized by performing an operation of calculating a drug evaluation system.

13. In paragraph 11, The above control unit, From the above PK profile, A 1-compartment model in which the natural logarithm of blood concentration over time is expressed as a straight line; and A drug evaluation system characterized in that the natural logarithm of blood concentration over time is determined by one of two-compartment models that is not expressed by a straight line.

14. In paragraph 11, The above control unit, The amount of drug to be administered into the microfluidic system from the determined compartment model (D system ) and a drug evaluation system characterized by determining the drug elimination rate constant k.

15. In paragraph 11, The above control unit, The amount of drug to be administered within the microfluidic system (D system ) is characterized by determining the following. D system = V system × C0 (Here, V system is the total volume of actual fluid in the microfluidic system, and C0 is the hyperconcentration, which is the blood concentration at time 0.) 16. In paragraph 11, The above control unit, For the 1-compartment model, the drug elimination rate constant k is determined by the following equation, k= {ln(C0) - ln(C t )} / t (Here, C0 is the initial concentration, which is the blood concentration at time 0, C t is the blood concentration at a specific time (t), where t is time.) From the above dissipation rate constant k, the liquid volume (V) that must be replaced per minute in the microfluidic system r,1min ) is determined by the following method. V r,1min = V system × (1 - e -k ) (Here, V system is the total volume of actual fluid in the microfluidic system, and k is the elimination rate constant of the drug.) 17. In paragraph 11, The above control unit, For the 2-compartment model, The vanishing velocity constant k1 in the first interval from time 0 to time t1; and The vanishing velocity constant k2 in the second section from time t1 to time t2 is determined by the following equation, k 1= {ln(C0)-ln(C1)} / (t1-0) k 2= {ln(C1)-ln(C2)} / (t2-t1) (Here, C0 is the initial concentration, C1 is the blood concentration at t1, and C2 is the blood concentration at t2.) The above dissipation rate constant k1 and The volume of liquid that must be replaced per minute in the microfluidic system from k2 (V r,1min ) is determined by the following method. V in the first section r,1min = V system × (1 - e -k1 ) V in the second section r,1min = V system × (1 - e -k2 ) (Here, V system is the total volume of the actual fluid in the microfluidic system, k1 is the disappearance rate constant of the drug in the first section, and k2 is the disappearance rate constant of the drug in the second section.) 18. In paragraph 11, The above control unit, For the 2-compartment model, The drug elimination rate constant k from two graphs using the residual method n Decide to do so, k n =ln{(Ae -αtn +Be -βtn ) / (Ae -αt(n+1) +Be -βt(n+1) )} (Here, k n is the decay rate constant at time n minutes, α is the slope of the first graph to which the residual method is applied, A is the y-intercept of the first graph, β is the slope of the second graph to which the residual method is applied, B is the y-intercept of the second graph, tn is time n, and tn+1 is time n+1.) The above k n Using the value, V at time n according to the following equation r,1min A drug evaluation system characterized by obtaining . V r,1min = V system × (1 - e -kn )

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