CN118197656A - Combined drug evaluation method based on three-dimensional human micro-physiological system - Google Patents
Combined drug evaluation method based on three-dimensional human micro-physiological system Download PDFInfo
- Publication number
- CN118197656A CN118197656A CN202410368467.5A CN202410368467A CN118197656A CN 118197656 A CN118197656 A CN 118197656A CN 202410368467 A CN202410368467 A CN 202410368467A CN 118197656 A CN118197656 A CN 118197656A
- Authority
- CN
- China
- Prior art keywords
- physiological
- organ
- dimensional
- human micro
- physiological system
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 81
- 229940079593 drug Drugs 0.000 title claims abstract description 57
- 238000011156 evaluation Methods 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 31
- 230000000694 effects Effects 0.000 claims abstract description 15
- 210000003484 anatomy Anatomy 0.000 claims abstract description 14
- 230000000857 drug effect Effects 0.000 claims abstract description 13
- 239000004615 ingredient Substances 0.000 claims abstract description 5
- 231100000048 toxicity data Toxicity 0.000 claims abstract description 4
- 210000000056 organ Anatomy 0.000 claims description 94
- 210000004027 cell Anatomy 0.000 claims description 63
- 239000011664 nicotinic acid Substances 0.000 claims description 41
- 230000001988 toxicity Effects 0.000 claims description 26
- 231100000419 toxicity Toxicity 0.000 claims description 26
- 230000002792 vascular Effects 0.000 claims description 22
- 208000031229 Cardiomyopathies Diseases 0.000 claims description 18
- 239000000890 drug combination Substances 0.000 claims description 15
- 230000008859 change Effects 0.000 claims description 14
- 230000001575 pathological effect Effects 0.000 claims description 12
- 239000008280 blood Substances 0.000 claims description 11
- 201000007270 liver cancer Diseases 0.000 claims description 11
- 208000014018 liver neoplasm Diseases 0.000 claims description 11
- 210000002569 neuron Anatomy 0.000 claims description 11
- 206010058467 Lung neoplasm malignant Diseases 0.000 claims description 10
- 230000008499 blood brain barrier function Effects 0.000 claims description 10
- 210000001218 blood-brain barrier Anatomy 0.000 claims description 10
- 201000005202 lung cancer Diseases 0.000 claims description 10
- 208000020816 lung neoplasm Diseases 0.000 claims description 10
- 230000006870 function Effects 0.000 claims description 9
- 210000002216 heart Anatomy 0.000 claims description 9
- 238000010146 3D printing Methods 0.000 claims description 8
- 239000012530 fluid Substances 0.000 claims description 8
- 210000004072 lung Anatomy 0.000 claims description 8
- 210000004556 brain Anatomy 0.000 claims description 7
- 210000004185 liver Anatomy 0.000 claims description 7
- 210000003734 kidney Anatomy 0.000 claims description 6
- 206010027476 Metastases Diseases 0.000 claims description 5
- 230000004888 barrier function Effects 0.000 claims description 5
- 230000009401 metastasis Effects 0.000 claims description 5
- 230000035790 physiological processes and functions Effects 0.000 claims description 5
- 239000005517 L01XE01 - Imatinib Substances 0.000 claims description 4
- 238000010276 construction Methods 0.000 claims description 4
- 210000002249 digestive system Anatomy 0.000 claims description 4
- KTUFNOKKBVMGRW-UHFFFAOYSA-N imatinib Chemical compound C1CN(C)CCN1CC1=CC=C(C(=O)NC=2C=C(NC=3N=C(C=CN=3)C=3C=NC=CC=3)C(C)=CC=2)C=C1 KTUFNOKKBVMGRW-UHFFFAOYSA-N 0.000 claims description 4
- 229960002411 imatinib Drugs 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 238000013528 artificial neural network Methods 0.000 claims description 3
- 229940000425 combination drug Drugs 0.000 claims description 3
- 238000005516 engineering process Methods 0.000 claims description 3
- 230000003902 lesion Effects 0.000 claims description 3
- 210000004324 lymphatic system Anatomy 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 239000000919 ceramic Substances 0.000 claims description 2
- 230000006854 communication Effects 0.000 claims description 2
- 238000004891 communication Methods 0.000 claims description 2
- 230000007721 medicinal effect Effects 0.000 claims description 2
- 239000011347 resin Substances 0.000 claims description 2
- 229920005989 resin Polymers 0.000 claims description 2
- 230000011218 segmentation Effects 0.000 claims description 2
- 238000012377 drug delivery Methods 0.000 claims 1
- 231100000331 toxic Toxicity 0.000 abstract description 2
- 230000002588 toxic effect Effects 0.000 abstract description 2
- 230000003993 interaction Effects 0.000 abstract 1
- AOJJSUZBOXZQNB-TZSSRYMLSA-N Doxorubicin Chemical compound O([C@H]1C[C@@](O)(CC=2C(O)=C3C(=O)C=4C=CC=C(C=4C(=O)C3=C(O)C=21)OC)C(=O)CO)[C@H]1C[C@H](N)[C@H](O)[C@H](C)O1 AOJJSUZBOXZQNB-TZSSRYMLSA-N 0.000 description 18
- 210000001519 tissue Anatomy 0.000 description 13
- 230000003592 biomimetic effect Effects 0.000 description 11
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 9
- 229960004316 cisplatin Drugs 0.000 description 9
- 229960004679 doxorubicin Drugs 0.000 description 9
- 238000010166 immunofluorescence Methods 0.000 description 9
- 206010028980 Neoplasm Diseases 0.000 description 8
- SNIOPGDIGTZGOP-UHFFFAOYSA-N Nitroglycerin Chemical compound [O-][N+](=O)OCC(O[N+]([O-])=O)CO[N+]([O-])=O SNIOPGDIGTZGOP-UHFFFAOYSA-N 0.000 description 7
- 239000000006 Nitroglycerin Substances 0.000 description 7
- 238000003501 co-culture Methods 0.000 description 7
- 229960003711 glyceryl trinitrate Drugs 0.000 description 7
- 201000011510 cancer Diseases 0.000 description 6
- 238000010586 diagram Methods 0.000 description 6
- 210000005166 vasculature Anatomy 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 5
- 210000004204 blood vessel Anatomy 0.000 description 5
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 210000001365 lymphatic vessel Anatomy 0.000 description 4
- 239000004005 microsphere Substances 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 210000005013 brain tissue Anatomy 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 230000003278 mimic effect Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000013334 tissue model Methods 0.000 description 3
- 210000000689 upper leg Anatomy 0.000 description 3
- XUKUURHRXDUEBC-KAYWLYCHSA-N Atorvastatin Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-KAYWLYCHSA-N 0.000 description 2
- XUKUURHRXDUEBC-UHFFFAOYSA-N Atorvastatin Natural products C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CCC(O)CC(O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-UHFFFAOYSA-N 0.000 description 2
- MLDQJTXFUGDVEO-UHFFFAOYSA-N BAY-43-9006 Chemical compound C1=NC(C(=O)NC)=CC(OC=2C=CC(NC(=O)NC=3C=C(C(Cl)=CC=3)C(F)(F)F)=CC=2)=C1 MLDQJTXFUGDVEO-UHFFFAOYSA-N 0.000 description 2
- 101000979001 Homo sapiens Methionine aminopeptidase 2 Proteins 0.000 description 2
- 101000969087 Homo sapiens Microtubule-associated protein 2 Proteins 0.000 description 2
- 239000005511 L01XE05 - Sorafenib Substances 0.000 description 2
- 102100021118 Microtubule-associated protein 2 Human genes 0.000 description 2
- 210000001015 abdomen Anatomy 0.000 description 2
- 229960001686 afatinib Drugs 0.000 description 2
- ULXXDDBFHOBEHA-CWDCEQMOSA-N afatinib Chemical compound N1=CN=C2C=C(O[C@@H]3COCC3)C(NC(=O)/C=C/CN(C)C)=CC2=C1NC1=CC=C(F)C(Cl)=C1 ULXXDDBFHOBEHA-CWDCEQMOSA-N 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 229940041181 antineoplastic drug Drugs 0.000 description 2
- 239000002473 artificial blood Substances 0.000 description 2
- 229960005370 atorvastatin Drugs 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000032677 cell aging Effects 0.000 description 2
- 210000000038 chest Anatomy 0.000 description 2
- 238000011260 co-administration Methods 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 210000000245 forearm Anatomy 0.000 description 2
- 210000003128 head Anatomy 0.000 description 2
- 210000001624 hip Anatomy 0.000 description 2
- 239000000017 hydrogel Substances 0.000 description 2
- 238000010874 in vitro model Methods 0.000 description 2
- 210000001699 lower leg Anatomy 0.000 description 2
- 210000004880 lymph fluid Anatomy 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 210000004165 myocardium Anatomy 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229960003787 sorafenib Drugs 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 206010013710 Drug interaction Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 206010050017 Lung cancer metastatic Diseases 0.000 description 1
- 208000029578 Muscle disease Diseases 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001601 blood-air barrier Anatomy 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000002932 cholinergic neuron Anatomy 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- NJDNXYGOVLYJHP-UHFFFAOYSA-L disodium;2-(3-oxido-6-oxoxanthen-9-yl)benzoate Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=CC(=O)C=C2OC2=CC([O-])=CC=C21 NJDNXYGOVLYJHP-UHFFFAOYSA-L 0.000 description 1
- 210000005064 dopaminergic neuron Anatomy 0.000 description 1
- 230000003291 dopaminomimetic effect Effects 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 238000001647 drug administration Methods 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 230000005713 exacerbation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000002483 medication Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 230000002107 myocardial effect Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000004126 nerve fiber Anatomy 0.000 description 1
- 210000003061 neural cell Anatomy 0.000 description 1
- 230000007243 organ organ communication Effects 0.000 description 1
- 210000002220 organoid Anatomy 0.000 description 1
- 238000005192 partition Methods 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 230000002633 protecting effect Effects 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 230000003362 replicative effect Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001578 tight junction Anatomy 0.000 description 1
- 231100000440 toxicity profile Toxicity 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of drug evaluation, in particular to a combined drug evaluation method based on a three-dimensional human micro-physiological system, which comprises the following steps: constructing a human micro-physiological system simulating a three-dimensional anatomical structure; applying one of n single pharmaceutical ingredients to the human micro-physiological system respectively; the human micro-physiological system is combined; and comparing the obtained drug effect and/or toxicity data to obtain the evaluation result of the combined drug. The invention can reveal the interaction mechanism of medicines possibly existing in the combined medicine and complications possibly caused, provides scientific reference for doctors to prescribe multiple medicines, greatly reduces the prescription error rate caused by empirical prescription, improves the treatment effect and welfare of patients, and simultaneously provides a method for comparing different medicine combinations and a method for comparing different curative effects and toxic effects of the same medicine combination on the elderly and normal people for medical researchers.
Description
Technical Field
The invention relates to the technical field of drug evaluation, in particular to a combined drug evaluation method based on a three-dimensional human micro-physiological system.
Background
In real life, many people often take multiple medicines simultaneously when suffering from multiple diseases, and this is especially common for the elderly. However, the simultaneous administration of multiple drugs may cause various problems, including unpredictable drug interactions and potential complications. For example, the use of anticancer drugs often may lead to complications such as cardiomyopathy. At present, an effective means for thoroughly solving the problem is not available. Physicians often prescribe a variety of medications through inquiry and clinical experience, and although this approach sometimes achieves a certain effect, it does not completely solve the above-mentioned problems. Clinicians may take incorrect prescriptions or incorrect doses, resulting in exacerbations or new complications for the patient. This situation not only affects the patient, but also places additional burden on the social healthcare system, and thus this problem has become a global challenge in nature.
The key to solving this problem is to scientifically evaluate the influence of simultaneous use of multiple drugs on human body. Firstly, the existing drug effect and toxicity evaluation standards are aimed at single drugs, and no evaluation paradigm is determined for combined administration; second, there is a lack of in vitro cell or tissue models for drug efficacy and toxicity evaluation of the combination, and thus no relevant clinical trials can be conducted. The existing human body imitated cell or tissue model has poor bionic degree, and a result with reference is difficult to obtain. In view of the complexity of multiple diseases and the potential complications of co-administration, the development of an advanced, highly biomimetic in vitro model capable of accurately replicating the complexity of the human body is critical for objective assessment of co-administration.
Disclosure of Invention
Therefore, the technical problem to be solved by the invention is to solve the problems that the existing drug effect and toxicity evaluation standards in the prior art are all aimed at single drugs, the combined drug has no definite evaluation paradigm, and the in-vitro high bionic cell or tissue model for evaluating the drug effect and toxicity of the combined drug is lacking.
In order to solve the technical problems, the invention provides a method for evaluating the combined medication by using a human micro-physiological system simulating a three-dimensional anatomical structure, which comprises the following steps:
S1, constructing a human body micro-physiological system simulating a three-dimensional anatomical structure; the human micro-physiological system comprises a three-dimensional bionic main body, a plurality of organ chambers correspondingly arranged in the three-dimensional bionic main body, and a vascular system for providing physiological support for bionic organs in the organ chambers, wherein the vascular system comprises a vascular system and/or a lymphatic system;
S2, respectively applying one of n single medicinal components to the human micro-physiological system, and measuring the medicinal effect and/or toxicity of the corresponding single medicinal component through the change of at least one bionic organ each time;
s3, carrying out combined medication on the human micro-physiological system, wherein the combined medication is obtained by combining n single drug components, and the drug effect and/or toxicity of the combined medication are determined through the change of at least one bionic organ;
s4, comparing the drug effect and/or toxicity data obtained by the S2 and the S3 to obtain an evaluation result of the combined drug.
In one embodiment of the present invention, in S1, the method for constructing the human micro-physiological system is as follows:
S11, obtaining a 3D printing electronic file of a bionic body based on a three-dimensional anatomical structure of a human body, and dividing the 3D printing electronic file into a plurality of functional module files, wherein a part of a vascular system and at least one organ cavity are arranged in each functional module file;
S12, performing 3D printing on the plurality of function module files generated by segmentation to obtain a plurality of function module entities;
S13, constructing a bionic organ in an organ cavity of the functional module entity through tissue and/or cell engineering technology;
s14, assembling a plurality of functional module entities into a complete three-dimensional bionic main body, and enabling vascular systems in the functional module entities to form a loop;
s15, enabling corresponding physiological liquid to circulate in the vascular system, and exciting physiological functions of the human micro-physiological system.
In one embodiment of the invention, in S5, each of said organ chambers is provided with at least one communication port for the construction of a biomimetic organ and/or for substance exchange with the vascular system.
In one embodiment of the present invention, in S6, the 3D printed material is selected from one or more of a resin, APL and ceramic.
In one embodiment of the present invention, in S3, further comprising: the method comprises the steps of grading the human micro-physiological system to implement the medicine component combination, wherein the grading implementation of the medicine component combination is to apply any combination from any two to any n-1 single medicine components, and each combination is used for measuring the medicine effect and/or toxicity of the medicine component combination through the change of at least one bionic organ.
In one embodiment of the invention, in S2, further comprising determining the efficacy and/or toxicity of the corresponding single pharmaceutical ingredient by a change in at least one physiological fluid; in S3, the method further comprises determining the efficacy and/or toxicity of the combination drug by a change in at least one physiological fluid, and determining the efficacy and/or toxicity of the combination of pharmaceutical ingredients by a change in at least one physiological fluid.
In one embodiment of the invention, the biomimetic organ comprises a normal organ as well as a pathological organ.
In one embodiment of the present invention, the normal organ comprises a brain including human neurons and blood brain barrier and a kidney including two-dimensionally cultured HK2 cells or three-dimensionally cultured HK2 cytoballs in a human micro-physiological system of a patient simulating liver cancer metastasis and lung cancer and having cardiomyopathy; the pathological organ comprises a liver, a lung and a heart in a human micro-physiological system of a patient with simulated liver cancer to metastasize lung cancer and cardiomyopathy, wherein the liver comprises two-dimensional HepG2 cells or three-dimensional cultured HepG2 cytoballs, and the lung comprises two-dimensional cultured A549 cells or three-dimensional cultured A549 cytoballs; the heart comprises two-dimensional cultured AC16 cells or three-dimensional cultured AC16 cell spheres of imatinib lesions.
In one embodiment of the invention, the biomimetic organ comprises cells with an age characteristic; the cells having age characteristics include cells in infant state, cells in toddler state, cells in young state, cells in adult state, and cells in elderly state.
In one embodiment of the invention, the human micro-physiological system further comprises a neural network providing physiological support for the organ chamber, a digestive system, a excretory system, a qi-blood barrier, and a blood-brain barrier.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
Firstly, the invention provides a reliable method for scientifically evaluating the effects of various drug combinations in human bodies, by simulating different pathological states in a three-dimensional human micro-physiological system and increasing the monomer number contained in the combined drug step by step, the drug effect and toxicity of the combined drug can be evaluated by accurately monitoring the change of physiological parameters of bionic organs or physiological fluids; the method can avoid the situation of prescribing a plurality of medicines only depending on doctor experience, so that medical researchers can comprehensively understand the characteristics of different medicine combinations, and provide scientific basis for formulating more effective treatment schemes, thereby improving the scientificity and accuracy of treatment, being beneficial to reducing adverse reactions and complications of medicines and improving the treatment effect and welfare of patients.
Secondly, the invention provides a method for scientifically comparing the effects of the same medicine combination on the old and young people, and the responses of different age groups to the medicine combination can be compared by simulating the physiological states of different age groups in a three-dimensional human body micro-physiological system and applying the same medicine combination; the method can help medical researchers to know the safety and effectiveness of the drug combination in different age groups, provides scientific basis for personalized treatment, and ensures the safety and effectiveness of combined administration in the elderly and young.
Drawings
In order that the invention may be more readily understood, a more particular description of the invention will be rendered by reference to specific embodiments thereof that are illustrated in the appended drawings, in which
FIG. 1 is a flow chart of a method for joint drug evaluation using a human micro-physiological system that simulates a three-dimensional anatomy;
FIG. 2 is a schematic diagram of the structure of a human micro-physiological system; wherein A is a human body st l file screenshot; b is a schematic diagram of the internal structure of a human body st l file; c is a module division diagram of the human micro-physiological system; d is an assembly schematic diagram of a human micro-physiological system;
FIG. 3A is a view showing an electronic file with a cut-out cross section; b is an image tightly connected by HUVEC; C. d is a representation of tight junctions of blood brain barrier cells (scale = 100 μm);
FIG. 4 is a schematic flow diagram of the vasculature in the human micro-physiological system;
FIG. 5A is a TH immunofluorescence image of the ibsc-dopaminergic neurons in Ge l MA; b is MAP2 immunofluorescence image of the ipsc-dopaminergic neuron in Ge l MA; c is CHAT immunofluorescence image of ipsc-cholinergic neurons in Ge l MA; d is MAP2 immunofluorescence image of the ibsc-cholinergic neurons in Ge l MA; e is CTNT immunofluorescence image of AC16 sphere; MRP2 immunofluorescence image of A549 sphere; g is an ALB immunofluorescence image of a HepG2 sphere; CK18 immunofluorescence image of HK2 sphere (scale bar = 100 μm);
Fig. 6a is a schematic diagram of a two-dimensional microchip structure for comparison in the third embodiment; b is a vigor comparison graph of co-culture of five primary tissues in a human micro-physiological system (three-dimensional human body chip), co-culture of two-dimensional human body chip and single culture;
FIG. 7 is a graph showing the results of toxicology and pharmacodynamics evaluation when three drugs based on the human micro-physiological system are combined;
FIG. 8 is a comparison of two prescriptions in example four, wherein A is CTNT immunofluorescence images of ac16 cells, positive control group induced with imatinib; b shows cell viability under both prescriptions; c shows cytotoxicity of both prescriptions to different cells;
FIG. 9 is a comparison of normal and aged human micro-physiological systems; wherein A is a display of cell aging; B. c, displaying multi-drug evaluation results based on the old human micro-physiological system and the normal human micro-physiological system;
FIG. 10 shows the fluorescence intensity measurements of sodium fluorescein transmitted by normal HUVEC and aged HUVEC over 24 h.
Detailed Description
The present invention will be further described with reference to the accompanying drawings and specific examples, which are not intended to be limiting, so that those skilled in the art will better understand the invention and practice it.
Example 1
Referring to fig. 1-4, the present invention employs an advanced biomimetic human micro-physiological system to simulate a three-dimensional anatomy of a human body, the system mainly comprising a three-dimensional biomimetic body, an organ chamber and a vascular system; the three-dimensional bionic body is constructed by a 3D printing technology or other manufacturing modes so as to simulate the whole structure of a human body; in some specific embodiments, the three-dimensional bionic body comprises a plurality of functional modules which can be assembled and disassembled, the functional modules comprise parts such as head, chest, waist and abdomen, thigh, shank, forearm and the like, each functional module forms the bionic body in a connection state, and organ culture or sampling operation can be respectively carried out on organ chambers in the functional modules in a disassembly state; a plurality of organ chambers are arranged in the three-dimensional bionic main body, each organ chamber simulates the position of an organ or tissue structure of a human body corresponding to the three-dimensional position, and the bionic organ is arranged in the organ chamber and is obtained by implantation or direct culture in the organ chamber; the bionic organ can be a normal organ or a pathological organ to simulate different physiological and pathological states in a human body; complex vascular systems including vascular systems and lymphatic systems are also built in the three-dimensional bionic body to simulate the blood circulation and immune system of a human body, and play a role in providing physiological support for bionic organs. By constructing the human micro-physiological system in S1, an in-vitro model simulating the complex structure and function inside the human body is established, and a reliable platform is provided for subsequent combined drug evaluation.
In S2, single drugs are applied to the human micro-physiological system in several times, usually, a single drug is dissolved or suspended in a physiological fluid at a proper concentration, and then the drug solution is injected into the human micro-physiological system by means of a syringe or a liquid inlet along with vascular system, etc., the physiological parameter changes of the bionic organ, such as the cell morphology, growth rate, metabolic activity, etc., are monitored in real time/at regular time, and the response of the bionic organ after each single drug is injected is recorded and analyzed to evaluate the drug effect and/or toxicity.
In S3, the combined drug to be tested formed by combining at least two single drugs is applied to the human micro-physiological system in a fractional manner, and the drug solution is applied and the drug effect and/or toxicity of the combined drug are evaluated in the same or similar way as that of S2; in S4, by comparing the drug effect and/or toxicity data obtained in the S2 and S3 stages, the evaluation result of the combined drug can be obtained, and the results can help the drug research and development personnel to know the comprehensive influence of different drug combinations on the human body, so that a reference basis is provided for the specification of the drug treatment scheme.
In one embodiment, in which the human micro-physiological system mimics a sample having liver cancer with metastatic lung cancer and having cardiomyopathy, the normal organs include normal brain and normal kidney, and the pathological organs include lung cancer lung, heart with cardiomyopathy and liver with liver cancer. Wherein, the normal brain comprises human neurons and blood brain barrier, and the normal kidney comprises two-dimensional culture HK2 cells or three-dimensional culture HK2 cell spheres; lung cancer lung comprises two-dimensional cultured A549 cells or three-dimensional cultured A549 cell spheres, heart muscle disease heart comprises two-dimensional cultured AC16 cells or three-dimensional cultured AC16 cell spheres damaged by Emartini, liver cancer liver comprises two-dimensional HepG2 cells or three-dimensional cultured HepG2 cell spheres.
In other specific embodiments, the biomimetic organ comprises cells with an age characteristic; the cells having age characteristics include cells in infant state, cells in toddler state, cells in young state, cells in adult state, and cells in elderly state. By constructing the bionic organ by using cells with different age characteristics, the physiological states of different age groups in a human body can be more truly simulated, so that the response of tissues with different age groups to the drug combination to be tested can be obtained, the drug effect and/or toxicity evaluation result of the drug combination to be tested based on the age characteristics can be further obtained, the accuracy and reliability of in-vitro drug evaluation can be improved, and more accurate guidance can be provided for clinical application.
In other specific embodiments, the human micro-physiological system further comprises a neural network that provides physiological support to the organ chamber, the digestive system, the excretory system, the qi-blood barrier, and the blood-brain barrier. The nerve cells in the nerve network sense the stimulus and transmit signals to other bionic organs or the whole system through nerve fibers to trigger corresponding physiological reactions so as to improve the simulation degree; when the medicine is injected into the organ cavity, the digestive system simulates the digestion process in the human body and decomposes and absorbs the medicine, thereby affecting the metabolism and action of the medicine; in the metabolic process of the excretory system, the excretory process in the human body is simulated, and the metabolic products are discharged out of the human body, so that the clearance of the medicine and the maintenance of steady-state concentration are influenced; the qi-blood barrier simulates the exchange process between blood and gas, affecting the distribution and transport of drugs in blood; the blood brain barrier simulates the exchange process between blood and brain tissue, limits certain substances from entering the brain tissue from the blood, protects the brain tissue from harmful substances, and reflects the distribution of the medicine in the brain more truly.
Example two
The present example provides the construction of a human micro-physiological system that mimics a cancer/cardiomyopathy patient:
The construction of the three-dimensional human micro-physiological system comprises several key steps: firstly, creating a digital st l file (3D printed file) of a human body, comprehensively referring to functional partitions of the human body and the convenience degree of assembly, cutting the file to obtain a plurality of functional modules (such as parts of head, chest, waist, abdomen, thigh, shank, forearm and the like), wherein each functional module can reflect an internal organ cavity (comprising a tissue cavity) and a vascular system, and the organ cavity in each functional module is customized according to a bionic organ or tissue, including pathological or aging organ and/or vascular morphology; then, the st l file is used for 3D printing to print out each entity function module; thereafter, a biomimetic organ is built within the organ chambers of the physical functional module by tissue and cell engineering techniques (the biomimetic organ is likewise customized), such as in combination with a culture medium (a culture medium liquid or hydrogel), in some cases a biomimetic organ or tissue may be built directly within a specific organ chamber (tissue lumen) or vasculature; then, assembling a plurality of entity function modules to form an artificial miniature or equal-proportion human body (three-dimensional bionic main body), and enabling vascular systems in the entity function modules to form a loop; after assembly, circulation of culture medium or other biological fluid is started in the system through the vascular system so as to activate the whole human micro-physiological system, so that the human micro-physiological system can act as a dynamic biological system; after the experiment is finished, separating each entity functional module, and extracting bionic organs (in organ chambers), tissues or body fluids from the entity functional modules for detection.
In a specific embodiment, a 3D three-dimensional human micro-physiological system (see fig. 2a, 2B, and 2C) with cancer metastasis and cardiomyopathy is designed and manufactured according to the above steps. The three-dimensional human micro-physiological system consists of a three-dimensional precisely positioned reflected sound "organ" chamber, "blood vessel" and "lymphatic vessel" (refer to D in figure 2). It contains five types of "organs" distributed in seven organ chambers, filled with Ge l MA hydrogels containing cells or cell aggregates. For example, dopaminergic and cholinergic neurons from human i PSC were contained in the head modular organ chamber, while lesion AC 16 microspheres (using imatinib as a damaging agent) simulating cardiomyopathy and a549 microspheres simulating lung cancer were included in the chest modular organ chamber (heart culture chamber, lung culture chamber). In addition, hepG2 microspheres are contained in the lumbar-abdominal modular organ chamber (liver-intestine culture chamber), representing liver cancer, and HK2 microspheres are contained in the thigh modular organ chamber (kidney culture chamber). The micro-channels represent blood and lymphatic vessels, which are responsible for the transport of artificial blood and lymphatic vessels, respectively. Simulating the scanning of a blood vessel from the top of an organ chamber, and simulating an air-blood barrier by clamping a porous membrane loaded with cells between the blood vessel and the organ; this embodiment simulates a lung cancer patient with liver cancer and cardiomyopathy metastasis, and when artificial blood and lymph fluid flow through the "human body", the three-dimensional human micro-physiological system is activated and physiological functions are developed.
To ensure accurate replication of the human body, it is important to establish efficient connections between different "organs" and their respective blood and lymphatic vessels in a three-dimensional human micro-physiological system. We have employed a technique whereby two vasculature passes through the top of an organ chamber, which can effectively facilitate organ-to-organ communication. The electronic file display shown in fig. 3a, which provides an interface between the organ and the vasculature, clearly shows the presence of two microchannels. Importantly, a porous membrane with HUVECs cells (see B in fig. 3) is placed between the organ chamber and the vasculature to ensure proper vascularization of the organ. Placed between the head module brain culture chamber and the vasculature is a porous membrane-based blood brain barrier (see C in fig. 3). The direction of blood and lymph fluid travel in a three-dimensional biomimetic human body is shown with reference to fig. 4.
In order to enhance the similarity of the three-dimensional human micro-physiological system to the actual human body, it is important to replicate the structure and function of the actual organs. In the field of drug discovery and precision medicine, a widely accepted method is to use cell aggregates such as organoids or cytospheroids as a mimic organ, and in this embodiment, a method of using cell aggregates as a mimic organ uses human cytospheroids in addition to ipsc-neurons and HUVECs cells to mimic organs (see a-H in fig. 5). In order to create an advantageous three-dimensional microenvironment for these cell aggregates or individual cells, ge l MA is used, a material widely used for culturing such cell structures.
Example III
The present embodiment provides characterization of three-dimensional human micro-physiological systems
To characterize the effects of co-culture of various organs in three-dimensional humans, more sensitive primary mouse tissues including brain, heart, kidney, lung and liver were used in this example. By respectively measuring the vitality after 15 days of co-culture of the three-dimensional human micro-physiological system, 15 days of co-culture of the two-dimensional human chips and 15 days of single culture, the vitality of the primary mouse tissues in the three-dimensional human micro-physiological system is found to be greater than that of the two-dimensional human chips and greater than that of the single culture after 15 days of co-culture; referring to fig. 6a, a two-dimensional human body chip manufactured according to chinese invention patent CN202211518830.4 as a control is shown; fig. 6B shows that the three-dimensional human micro-physiological system has better co-culture effect, can keep tissue activity with high fidelity, has high bionic property, and is suitable for multi-disease research.
Example IV
The embodiment provides the application of the three-dimensional human micro-physiological system in the multi-disease research
Cancer/cardiomyopathy is a common complication. In many cases, cardiomyopathy is caused directly by cancer treatment. This form of multipathogenic treatment becomes more challenging when cancer metastasis is involved, as many types of anticancer drugs are often involved. In the second embodiment, a three-dimensional human micro-physiological system simulating patients with cancers and cardiomyopathy is constructed, while in the present embodiment, the use of the three-dimensional human micro-physiological system to obtain the therapeutic effect and toxicity profile of the drug combination is shown, and scientific data is provided for doctors to make clinical decisions in treating cancers and cardiomyopathy. Specifically, a combination drug comprising doxorubicin, cisplatin and nitroglycerin is selected, wherein the cisplatin is aimed at lung cancer, the doxorubicin is aimed at liver cancer, and the nitroglycerin is aimed at cardiomyopathy. First, the efficacy and toxicity of a single component across an "organ" drug combination was measured, then the efficacy and toxicity of a two drug combination was measured, and finally the efficacy and toxicity of a three drug combination was measured. Through the comparison of the data, the characteristic of combined drug administration of the doxorubicin, the cisplatin and the nitroglycerin can be obtained.
Referring to fig. 7, the dose-response curves for doxorubicin, cisplatin and nitroglycerin were found to have half-inhibition concentrations at which they were allowed to act on other organs. It can be seen that doxorubicin and cisplatin have a variable inhibitory effect on other cells, except neural cells. The nerve cells have blood brain barrier, so the medicine can hardly act on the nerve cells, and the nerve cells keep high activity. When the drugs are mixed pairwise, the heart protecting effect of the nitroglycerin and the toxicity of the cisplatin and the doxorubicin to the cardiac muscle are opposite, so that the activity of myocardial cells is kept high, and when the cisplatin and the doxorubicin are mixed, the expected synergistic effect does not occur, which suggests that the drug combination is probably not optimal under our experimental conditions, but rather the synergistic effect is reflected after the three drugs are mixed. These results show that when various drugs are mixed, it is necessary to test the toxicity and efficacy curves in advance.
In addition to efficacy and toxicity, the three-dimensional human micro-physiological system can also be used to compare the effects of different drug combinations and the same drug combinations on normal and aged three-dimensional human micro-physiological systems. Firstly, evaluating two model drug combinations by using a normal three-dimensional human micro-physiological system, and evaluating the curative effect and safety of the drug combinations. Prescription 1 is "doxorubicin, cisplatin and nitroglycerin" and prescription 2 is "afatinib, sorafenib and atorvastatin". Specifically, cisplatin and afatinib are directed against lung cancer, doxorubicin and sorafenib are directed against liver cancer, nitroglycerin and atorvastatin are directed against cardiomyopathy. Referring to FIG. 8A, the efficacy of formulation-1 on injured AC-16 was significantly changed from the positive control, while formulation-2 was not significantly changed. Referring to FIG. 8B, it is evident that prescription-1 has a similar inhibitory effect on HepG2 and slightly stronger inhibitory effect on A549. In addition, it is more toxic to HK2, AC16 than prescription 2. The results shown in fig. 8B and fig. 8C are identical. Based on these results, it can be concluded that prescription-2 is better suited than prescription-1 to address the co-occurrence of cancer and cardiomyopathy.
Next, the efficacy and safety of the formulation-2 in the normal three-dimensional human micro-physiological system and the three-dimensional micro-physiological system of the aged human with cell aging can be compared (refer to a shown in fig. 9 a). Referring to FIG. 9B, the survival rate of HK2 in the three-dimensional human micro-physiological system of the elderly is approximately half that of the normal three-dimensional human micro-physiological system, indicating that the safe dose of prescription 2 to the elderly should be significantly lower than for adults. Interestingly, referring to FIG. 9C, the cytotoxicity of prescription-2 to HepG2, A549 and AC16 showed about 5% difference between normal and aged mps, although these pathological "organs" were essentially identical. The slight change can be caused by two reasons, namely, firstly, the HK2 of the three-dimensional human micro-physiological system of the old undergoes massive apoptosis, and a large amount of inflammatory factors are released, so that other pathological organs are influenced; second, aged HUVECs lead to increased barrier permeability (see fig. 10), leading to higher drug concentrations interacting with pathological "organs" within the system.
It is apparent that the above examples are given by way of illustration only and are not limiting of the embodiments. Other variations and modifications of the present invention will be apparent to those of ordinary skill in the art in light of the foregoing description. It is not necessary here nor is it exhaustive of all embodiments. And obvious variations or modifications thereof are contemplated as falling within the scope of the present invention.
Claims (10)
1. A method for evaluating combined medication by using a human micro-physiological system simulating a three-dimensional anatomical structure is characterized by comprising the following steps:
S1, constructing a human body micro-physiological system simulating a three-dimensional anatomical structure; the human micro-physiological system comprises a three-dimensional bionic main body, a plurality of organ chambers correspondingly arranged in the three-dimensional bionic main body, and a vascular system for providing physiological support for bionic organs in the organ chambers, wherein the vascular system comprises a vascular system and/or a lymphatic system;
S2, respectively applying one of n single medicinal components to the human micro-physiological system, and measuring the medicinal effect and/or toxicity of the corresponding single medicinal component through the change of at least one bionic organ each time;
s3, carrying out combined medication on the human micro-physiological system, wherein the combined medication is obtained by combining n single drug components, and the drug effect and/or toxicity of the combined medication are determined through the change of at least one bionic organ;
s4, comparing the drug effect and/or toxicity data obtained by the S2 and the S3 to obtain an evaluation result of the combined drug.
2. The method for performing drug combination evaluation by using the human micro-physiological system simulating the three-dimensional anatomical structure according to claim 1, wherein in S1, the method for constructing the human micro-physiological system is as follows:
S11, obtaining a 3D printing electronic file of a bionic body based on a three-dimensional anatomical structure of a human body, and dividing the 3D printing electronic file into a plurality of functional module files, wherein a part of a vascular system and at least one organ cavity are arranged in each functional module file;
S12, performing 3D printing on the plurality of function module files generated by segmentation to obtain a plurality of function module entities;
S13, constructing a bionic organ in an organ cavity of the functional module entity through tissue and/or cell engineering technology;
s14, assembling a plurality of functional module entities into a complete three-dimensional bionic main body, and enabling vascular systems in the functional module entities to form a loop;
s15, enabling corresponding physiological liquid to circulate in the vascular system, and exciting physiological functions of the human micro-physiological system.
3. A method of constructing a human micro-physiological system according to claim 2, wherein in S5 each of said organ chambers is provided with at least one communication port for the construction of a bionic organ and/or for substance exchange with the vascular system.
4. The method of constructing a human micro-physiological system according to claim 2, wherein in S6, the 3D printed material is selected from one or more of resin, APL and ceramic.
5. The method for joint drug evaluation using a human micro-physiological system simulating three-dimensional anatomy according to claim 1, further comprising, in S3: the method comprises the steps of grading the human micro-physiological system to implement the medicine component combination, wherein the grading implementation of the medicine component combination is to apply any combination from any two to any n-1 single medicine components, and each combination is used for measuring the medicine effect and/or toxicity of the medicine component combination through the change of at least one bionic organ.
6. The method for combined use evaluation of human micro-physiological systems simulating three-dimensional anatomy according to any one of claims 1,2 or 5, further comprising determining the efficacy and/or toxicity of the corresponding single pharmaceutical ingredient by a change in at least one physiological fluid in S2; in S3, the method further comprises determining the efficacy and/or toxicity of the combination drug by a change in at least one physiological fluid, and determining the efficacy and/or toxicity of the combination of pharmaceutical ingredients by a change in at least one physiological fluid.
7. The method for combined drug delivery evaluation using a human micro-physiological system simulating three-dimensional anatomy according to claim 1, wherein the bionic organ includes a normal organ and a pathological organ.
8. The bionic organ according to claim 7, wherein the normal organ comprises brain including human neurons and blood brain barrier and kidney including two-dimensionally cultured HK2 cells or three-dimensionally cultured HK2 cell balls in a human micro-physiological system of a patient who simulates liver cancer metastasis and lung cancer and has cardiomyopathy; the pathological organ comprises a liver, a lung and a heart in a human micro-physiological system of a patient with simulated liver cancer to metastasize lung cancer and cardiomyopathy, wherein the liver comprises two-dimensional HepG2 cells or three-dimensional cultured HepG2 cytoballs, and the lung comprises two-dimensional cultured A549 cells or three-dimensional cultured A549 cytoballs; the heart comprises two-dimensional cultured AC16 cells or three-dimensional cultured AC16 cell spheres of imatinib lesions.
9. The method for joint drug use evaluation using a human micro-physiological system simulating three-dimensional anatomy according to claim 1, wherein the bionic organ includes cells with age characteristics; the cells having age characteristics include cells in infant state, cells in toddler state, cells in young state, cells in adult state, and cells in elderly state.
10. The method of claim 1, wherein the human micro-physiological system further comprises a neural network, a digestive system, a excretory system, a qi-blood barrier, and a blood-brain barrier that provide physiological support to the organ chamber.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410368467.5A CN118197656A (en) | 2024-03-28 | 2024-03-28 | Combined drug evaluation method based on three-dimensional human micro-physiological system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410368467.5A CN118197656A (en) | 2024-03-28 | 2024-03-28 | Combined drug evaluation method based on three-dimensional human micro-physiological system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN118197656A true CN118197656A (en) | 2024-06-14 |
Family
ID=91400027
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410368467.5A Pending CN118197656A (en) | 2024-03-28 | 2024-03-28 | Combined drug evaluation method based on three-dimensional human micro-physiological system |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN118197656A (en) |
-
2024
- 2024-03-28 CN CN202410368467.5A patent/CN118197656A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Goriely et al. | Mechanics of the brain: perspectives, challenges, and opportunities | |
| Bae et al. | Advanced drug delivery 2020 and beyond: Perspectives on the future | |
| Zhu et al. | 3D printing promotes the development of drugs | |
| Selimović et al. | Organs-on-a-chip for drug discovery | |
| Beecher | Experimental pharmacology and measurement of the subjective response | |
| Bhuskute et al. | 3D printed personalized medicine for cancer: applications for betterment of diagnosis, prognosis and treatment | |
| Baum et al. | Role of complementary and alternative medicine in the care of patients with breast cancer: report of the European Society of Mastology (EUSOMA) Workshop, Florence, Italy, December 2004 | |
| US20170157318A1 (en) | System and Treatment Method to Increase Circulation and Pluripotency of Stem and Progenitor Cells within a Patient | |
| O’farrell et al. | Luminal fluid motion inside an in vitro dissolution model of the human ascending colon assessed using magnetic resonance imaging | |
| RU2584598C1 (en) | Microfluid chip for creation of mammalian organ cell models | |
| Cho et al. | High-throughput zebrafish intramuscular recording assay | |
| CN118197656A (en) | Combined drug evaluation method based on three-dimensional human micro-physiological system | |
| Lozoya-Agullo et al. | Preclinical models for colonic absorption, application to controlled release formulation development | |
| Stefanov et al. | Western medicine versus Eastern medicine: do both have a common root, scientific background, and worldwide recognition | |
| Zhang | A medical mini-me one day your doctor could prescribe drugs based on now a biochip version of you reacts | |
| Roldan et al. | Head phantom for the acquisition of pulsatile optical signals for traumatic brain injury monitoring | |
| Narasimhan | The role of porous medium modeling in biothermofluids | |
| Poorman et al. | Naturopathic medical education: where conventional, complementary, and alternative medicine meet | |
| Menshykau et al. | Mechanistic image-based modelling: concepts and applications | |
| Kumari et al. | IMPORTANCE OF DASHVIDHA ATUR PARIKSHA IN CLINICAL PRACTICE | |
| Chandrakar et al. | Impact of Perfusion on Growth and Drug Delivery in Vascularized Tumor Spheroid-On-A-Chip Models | |
| Hansdah et al. | A Concept of Paradi Guna (Samyoga) and its Clinical Utility | |
| Convener | In association with | |
| Walmstedt | Mathematical modelling of microvascular oxygen transport and the impact of blood pressure | |
| Farooqi | Effect of Methanolic Extracts of Acer Cappadocicum on HepG2 Cancer Cell Line in a Liver Microphysiological System |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination |