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Identifying compounds modifying a cellular phenotype

a technology of cellular phenotype and compound, applied in the field of identifying compounds modifying cellular phenotype, can solve the problems of frequent limited access to primary cells and susceptible primary cells, and achieve the effect of simple operation for the user

Inactive Publication Date: 2019-11-21
2CUREX APS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This method enables efficient testing of large panels of compounds on limited cell samples with minimal handling, accurately recapitulating in vivo behavior, thereby identifying effective treatments for cancer patients.

Problems solved by technology

Primary cells are susceptible, and should therefore only be subjected to as few handling steps as possible.
Furthermore, access to primary cells is frequently limited and in order to avoid unnecessary loss of cell material as few handling steps as possible are desirable.

Method used

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  • Identifying compounds modifying a cellular phenotype
  • Identifying compounds modifying a cellular phenotype
  • Identifying compounds modifying a cellular phenotype

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0329]Loading of Array with Chemo-Therapeutic Compounds “Freely” Distributed in the Support as Single Compounds and in Combinations at Various Concentrations and Ratios

Materials and Solutions

[0330]Support: Matrigel® available from BD Biosciences.[0331]Arrays: Micro slide format arrays with 96 individual wells surrounded by a common reservoir. Each well has a volume of 20 μL and the distance between neighboring wells was 1 mm.[0332]Chemo therapeutic compounds (Irinitecan, Oxaliplatin, Leucovorin, 5FU, Cetuximab)[0333]Cooling device for keeping 5° C.

Procedure:

[0334]Compounds, Matrigel® and arrays are kept at 5° C. (cooling device) throughout the process when not in the freezer.[0335]1. Compounds were diluted in Matrigel® (Fluid) as single compounds and / or in combinations equal to standard treatment regimens of patients with colorectal cancer.[0336]2. Matrigel® with compounds were added to individual wells of the arrays (Fluid)[0337]3. Arrays were centrifuged 10 min at 3000 RPM (cold)[...

example 2

[0339]Loading of Arrays with Test Compounds Imbedded in Thermo Sensitive Nano-Particles Distributed in Matrigel; for Release Upon Demand

Materials and Solutions:

[0340]Nano-particles containing chemo therapeutic compounds[0341]Support: Matrigel® available from BD Biosciences,[0342]Arrays: Micro slide format array with 96 individual wells surrounded by a common reservoir as described in Example 1[0343]Cooling device for keeping 5° C.

Procedure:

[0344]Nano-particles, Matrigel® and arrays were kept at 5° C. (culling block) throughout the process when not in the freezer.[0345]1. Compound containing nano-particles were diluted in Matrigel® (fluid).[0346]2. Matrigel containing the nano-particles (from above point 1) were added to individual wells of the array[0347]3. The arrays were centrifuged 10 min at 3000 RPM (cold)[0348]4. The arrays were stored at −18° C. for 24 hrs. (solid)

example 3

[0349]No Spill Over Occurs Between Wells Because the Support is Solid then Cells are Added to the Reservoir

Materials and Solutions

[0350]Support: Matrigel® available from BD Biosciences[0351]Arrays: Micro slide format arrays with 96 individual s surrounded by a common reservoir as described in Example 1[0352]Chemo therapeutic compound: Doxorubicin[0353]Cooling device for keeping 5° C.

Procedure:

[0354]Compounds, Matrigel® and arrays are kept at 5° C. (cooling device) t1 throughout the process when not in the freezer.[0355]1. Doxorubicin was diluted in Matrigel® (Fluid)[0356]2. Matrigel® with Doxorubicin was added to individual wells of the arrays (Fluid)[0357]3. The array was centrifuged 10 min at 3000 RPM (cold)[0358]4. The array was stored at −18° C. for 24 hrs. (Solid)[0359]1. Tumor tissue from patient was prepared to form a population of small individual micro-tumors (spheroids)[0360]2. Micro-tumors were suspended in cold STEM media[0361]3. The array was taken directly from freezer...

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Abstract

The present invention relates to a method and tools for extracting information on a compounds influence on a cellular phenotype. The method of the invention may be used as a very efficient procedure for testing the efficacy or resistance of single drugs or combinations of drugs on cells from individual patients. Thus, the methods may be useful for predicting efficacy of a drug on a given patient. The methods are also useful for testing of compounds for toxicity, identifying drug targets for known or novel compounds.

Description

REFERENCE TO RELATED APPLICATIONS[0001]This application is continuation of U.S. patent application Ser. No. 15 / 321,888, filed Dec. 23, 2016, which is a U.S. national stage application of PCT / DK2015 / 050197, filed Jul. 1, 2015, which claims priority to Denmark Patent Application No. PA201400357, filed Jul. 2, 2014. The entire content of each application is incorporated herein by reference.FIELD OF THE INVENTION[0002]The present invention relates to a method and tools for extracting information relating to the influence on a cellular phenotype or cellular property influenced by a compound or a combination of compounds. In particular, the method relates to a semi solid support that through multiple phase shifts allow loading of compounds and cells into the support and testing the influence of said compounds on the cellular system.[0003]The method of the invention may be used as a very efficient procedure for testing or discovering the influence of a library of compounds on a cellular pr...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/50C07K16/28A61K31/704A61K31/555A61K31/513A61K31/4745A61K31/519
CPCA61K31/555A61K31/4745A61K31/513G01N33/5008A61K31/704G01N33/5011C07K16/2863A61K31/519
Inventor HAGEL, GRITHTHASTRUP, OLE
Owner 2CUREX APS
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