WO2020172337A1 - Methods to predict repeated dose toxicity using an integrated organ platform - Google Patents

Methods to predict repeated dose toxicity using an integrated organ platform Download PDF

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Publication number
WO2020172337A1
WO2020172337A1 PCT/US2020/018911 US2020018911W WO2020172337A1 WO 2020172337 A1 WO2020172337 A1 WO 2020172337A1 US 2020018911 W US2020018911 W US 2020018911W WO 2020172337 A1 WO2020172337 A1 WO 2020172337A1
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WO
WIPO (PCT)
Prior art keywords
toxicity
tissue
methods
predict
repeated dose
Prior art date
Application number
PCT/US2020/018911
Other languages
French (fr)
Inventor
James Mckim
Original Assignee
Iontox, Llc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Iontox, Llc filed Critical Iontox, Llc
Priority to EP20759516.6A priority Critical patent/EP3927417A4/en
Publication of WO2020172337A1 publication Critical patent/WO2020172337A1/en

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity

Definitions

  • the present invention is in the field of toxicity testing for various pharmaceuticals.
  • a method of testing the toxicity of a compound to a specific tissue comprising: growing the tissue three dimensionally in an integrated plate system, contacting the tissue with the compound; and measuring the toxicity of the compound on the tissue.
  • Figure 1 shows an integrated human organ plate with simulated blood flow.
  • Figure 2 shows a diagrammatic side view of three organ integrated plate.
  • Figure 3 shows the data for determining acetaminophen pharmacokinetic s .
  • Figure 4 shows the data for gene expression to predict toxicity.
  • Figure 5 shows the data for cyclohexamide pharmacokinetics in the HuDMOP system.
  • Figure 6 shows the data for cyclohexamide cytotoxicity in three organs.
  • Figure 7A shows the data for the skin model with scopalmine.
  • Figure 7B shows the data for the liver model with scopalmine.
  • Three-dimensionally grown human tissue e.g., intestinal tissue, liver tissue, and kidney tissue, were established in an integrated plate format with simulated blood.
  • An example of the integrated plate format is disclosed in the U.S. Application Publication 2015/0267158, incorporated herein by reference in its entirety, including the drawings.
  • Test materials were then added to the apical side of the tissue model at time 0. Dose selection was based on in vitro studies to determine the maximum tolerated dose level (MTD).
  • the simulated blood flow pumps were started, and samples were collected at regular time intervals up to 24 hr. Samples were taken from the various tissue compartments, e.g., the basolateral intestinal compartment, the liver media compartment, the renal compartment and from the simulated blood. At 24 hr a second dose was applied to the apical surface of the intestine tissue and samples were again collected over the next 24 hr at regular intervals.
  • the three-organ plate set is shown in Figure 1. A side view showing the various sampling compartments is shown in Figure 2.
  • Cm x for multiple drugs correlated well with the maximum concentration in the basolateral space (simulating systemic interstitial space), The deflection in the simulated blood curve supported this as does the time to maximum (Tmax). Both the Cm x and Tmax values in the plate system matched clinical data for the drugs. An example of these kinetics are shown with the drug acetaminophen in Figure 3.
  • Systemic organ specific toxicity was predicted by using a custom algorithm that incorporated the kinetic data as well as key biochemical and molecular markers. Specifically, these markers included Cytochrome P450 induction, Nrf2 activation, and key Nrf2 controlled genes (See Figure 4).
  • cycloheximide is a potent inhibitor of protein synthesis and highly toxic.
  • the pharmacokinetic data in Figure 6 demonstrated absorption, Cm x and Tm x all of which were consistent with clinical data.
  • the toxicity markers indicated key genes were turned on and that toxicity occured in the intestine and liver. Again, these results were consistent with known animal and human clinical data.
  • This approach can also be used with drugs that are aerosols, vapors, or gases by using an inhalation chamber that attaches to the HuDMOP plate. This yielded a lung-liver-kidney model. Another iteration was to use skin-liver-kidney (see Figure 5).

Abstract

A method of testing the toxicity of a compound to a specific tissue, the method comprising: growing the tissue three dimensionally in an integrated plate system, contacting the tissue with the compound; and measuring the toxicity of the compound on the tissue.

Description

METHODS TO PREDICT REPEATED DOSE TOXICITY
USING AN INTEGRATED ORGAN PLATFORM
RELATED APPLICATION
[001] The present application claims priority to the U.S. Provisional Application Serial No. 62/807,708, filed on February 19, 2019 by James MCKIM, and entitled: “METHODS TO PREDICT REPEATED DOSE TOXICITY USING AN INTEGRATED ORGAN PLATFORM,” the entire disclosure of which is hereby incorporated herein by reference, including all the figures.
FIELD OF THE INVENTTION
[002] The present invention is in the field of toxicity testing for various pharmaceuticals.
BACKGROUND OF THE DISCLOSURE
[003] Currently drugs and chemicals are tested for toxicity in 14-, 28-, and 90-day studies using rats and dogs as surrogates for humans. The data obtained includes pharmacokinetic information, organ toxicity, metabolism and the development of dose- response relationships.
[004] There are currently no alternative (e.g., ex vivo, in vitro) methods to obtain these parameters. Therefore, a method that uses human tissues in an integrated platform that can provide the parameters required would be of great value to the pharmaceutical and chemical industries.
SUMMARY OF THE INVENTION
[005] A method of testing the toxicity of a compound to a specific tissue, the method comprising: growing the tissue three dimensionally in an integrated plate system, contacting the tissue with the compound; and measuring the toxicity of the compound on the tissue.
BRIEF DISCRETION OF THE DRAWINGS
[006] Figure 1 shows an integrated human organ plate with simulated blood flow. [007] Figure 2 shows a diagrammatic side view of three organ integrated plate.
[008] Figure 3 shows the data for determining acetaminophen pharmacokinetic s .
[009] Figure 4 shows the data for gene expression to predict toxicity.
[0010] Figure 5 shows the data for cyclohexamide pharmacokinetics in the HuDMOP system.
[0011] Figure 6 shows the data for cyclohexamide cytotoxicity in three organs.
[0012] Figure 7A shows the data for the skin model with scopalmine.
[0013] Figure 7B shows the data for the liver model with scopalmine.
DETAILED DESCRIPTION OF THE EMBODIMENTS
[0014] Using the methods described herein, the inventors of the present disclosure were able to test the toxicity of various compounds in various tissues without the need to dose and animal. The dosing can be done at various time intervals that is much more rapid than in animal studies. The methods disclosed herein lead to faster, more accurate, and less expensive means of obtaining toxicity data.
[0015] Three-dimensionally grown human tissue, e.g., intestinal tissue, liver tissue, and kidney tissue, were established in an integrated plate format with simulated blood. An example of the integrated plate format is disclosed in the U.S. Application Publication 2015/0267158, incorporated herein by reference in its entirety, including the drawings.
[0016] Test materials were then added to the apical side of the tissue model at time 0. Dose selection was based on in vitro studies to determine the maximum tolerated dose level (MTD). The simulated blood flow pumps were started, and samples were collected at regular time intervals up to 24 hr. Samples were taken from the various tissue compartments, e.g., the basolateral intestinal compartment, the liver media compartment, the renal compartment and from the simulated blood. At 24 hr a second dose was applied to the apical surface of the intestine tissue and samples were again collected over the next 24 hr at regular intervals. The three-organ plate set is shown in Figure 1. A side view showing the various sampling compartments is shown in Figure 2.
[0017] Cm x for multiple drugs correlated well with the maximum concentration in the basolateral space (simulating systemic interstitial space), The deflection in the simulated blood curve supported this as does the time to maximum (Tmax). Both the Cm x and Tmax values in the plate system matched clinical data for the drugs. An example of these kinetics are shown with the drug acetaminophen in Figure 3.
[0018] Systemic organ specific toxicity was predicted by using a custom algorithm that incorporated the kinetic data as well as key biochemical and molecular markers. Specifically, these markers included Cytochrome P450 induction, Nrf2 activation, and key Nrf2 controlled genes (See Figure 4).
[0019] The result was that by using multiple dosing in vitro (optimized for 48 hr) the duration of studies were shortened, dose response curves were developped, metabolism and kinetic parameters were better understood, and organ specific toxicity was predicted. Absorption and bioavailability were also predicted accurately with this model system. These three organs were selected because they are the primary reasons for toxicity and they control 90% of drug or chemical pharmacokinetics. In the Acetaminopen (APAP) example below there was no significant toxicity. Only slight toxicity measured in liver and kidney. These data also corresponded to in vivo human clinical data.
[0020] In contrast to APAP, a non-toxic and safe drug, cycloheximide is a potent inhibitor of protein synthesis and highly toxic. The pharmacokinetic data in Figure 6 demonstrated absorption, Cm x and Tm x all of which were consistent with clinical data. The toxicity markers (see Figure 7) indicated key genes were turned on and that toxicity occured in the intestine and liver. Again, these results were consistent with known animal and human clinical data.
[0021] This approach can also be used with drugs that are aerosols, vapors, or gases by using an inhalation chamber that attaches to the HuDMOP plate. This yielded a lung-liver-kidney model. Another iteration was to use skin-liver-kidney (see Figure 5).

Claims

WHAT IS CLAIMED IS:
1. A method of testing the toxicity of a compound to a specific tissue, the method comprising:
growing the tissue three dimensionally in an integrated plate system, and contacting the tissue with the compound; and
measuring the toxicity of the compound on the tissue.
PCT/US2020/018911 2019-02-19 2020-02-19 Methods to predict repeated dose toxicity using an integrated organ platform WO2020172337A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP20759516.6A EP3927417A4 (en) 2019-02-19 2020-02-19 Methods to predict repeated dose toxicity using an integrated organ platform

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201962807708P 2019-02-19 2019-02-19
US62/807,708 2019-02-19

Publications (1)

Publication Number Publication Date
WO2020172337A1 true WO2020172337A1 (en) 2020-08-27

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EP (1) EP3927417A4 (en)
WO (1) WO2020172337A1 (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050260745A1 (en) * 2004-05-19 2005-11-24 Massachusetts Institute Of Technology Perfused three-dimensional cell/tissue disease models
US20060019326A1 (en) * 2003-01-16 2006-01-26 Vacanti Joseph P Use of three-dimensional microfabricated tissue engineered systems for pharmacologic applications
US20070099294A1 (en) * 2005-11-02 2007-05-03 The Ohio State University Research Foundation Materials and methods for cell-based assays
US20130143230A1 (en) * 2011-12-02 2013-06-06 The Trustees Of The Stevens Institute Of Technology Microfluidic-based cell-culturing platform and method
US20140030752A1 (en) * 2012-07-25 2014-01-30 Massachusetts Institute Of Technology Modular platform for multi-tissue integrated cell culture
WO2016100227A1 (en) * 2014-12-15 2016-06-23 The Regents Of The University Of California Multi-organ cell culture system and methods of use thereof

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9631167B2 (en) * 2014-03-21 2017-04-25 Iontox, Llc Dynamic multi organ plate
EP3940059A1 (en) * 2016-08-25 2022-01-19 Philip Morris Products S.A. Cell culture

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060019326A1 (en) * 2003-01-16 2006-01-26 Vacanti Joseph P Use of three-dimensional microfabricated tissue engineered systems for pharmacologic applications
US20050260745A1 (en) * 2004-05-19 2005-11-24 Massachusetts Institute Of Technology Perfused three-dimensional cell/tissue disease models
US20070099294A1 (en) * 2005-11-02 2007-05-03 The Ohio State University Research Foundation Materials and methods for cell-based assays
US20130143230A1 (en) * 2011-12-02 2013-06-06 The Trustees Of The Stevens Institute Of Technology Microfluidic-based cell-culturing platform and method
US20140030752A1 (en) * 2012-07-25 2014-01-30 Massachusetts Institute Of Technology Modular platform for multi-tissue integrated cell culture
WO2016100227A1 (en) * 2014-12-15 2016-06-23 The Regents Of The University Of California Multi-organ cell culture system and methods of use thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3927417A4 *

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Publication number Publication date
EP3927417A4 (en) 2022-11-16
EP3927417A1 (en) 2021-12-29

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