System and method for high throughput screening of cancer cells

A cancer cell, high-throughput technology, applied in compound screening/testing, measuring devices, instruments, etc., can solve long-standing and unsatisfied problems

Inactive Publication Date: 2018-05-11
CANNABICS PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In view of the above, there remains a long-standing and unmet need for the use of novel therapies such as the use of plant extracts to treat multifunctional diseases such as cancer

Method used

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  • System and method for high throughput screening of cancer cells
  • System and method for high throughput screening of cancer cells
  • System and method for high throughput screening of cancer cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0210] Cannabis Extraction Protocols

[0211] Dried flowers of six aged cannabis strains (i.e., CNB1, CNB2, CNB3, CNB4, CNB6, CNB8) were steeped in butane and purified with resin to exclude butane residues from the concentrated oil.

[0212] The concentrations of 10 cannabinoids (i.e., CBDA, CBGA, CBG, CBD, THCV, CBN, THCA, A9THC, A8THC, and CBC) in the extract were assessed using HPLC (see figure 1 ).

[0213] To prepare the stock for the cell lines, cannabis extract was dissolved in DMSO (eg about 50 mg / ml) and kept at -20°C until use.

Embodiment 2

[0215] Protocol for Cell Line Preparation

[0216] Referring now to non-limiting examples of cell cultures useful in the present invention:

[0217] Examples of Human Cancer Cell Lines

[0218] MDA-MB-231 and MCF-7 breast cancer cells

[0219] U87MG and U118MG glioblastoma cells

[0220] · PC3 prostate cancer cells

[0221] · HT29 and SW480 colon cancer cells

[0222] · AGS gastric adenocarcinoma

[0223] · MiaPaCa2 pancreatic cancer cells

[0224] Examples of human cancer cell lines (control group):

[0225] Stable non-tumor cell line HDF (human dermal fibroblast)

[0226] HaCat (human keratinocytes)

[0227] Now refer to the protocol for growing cell lines:

[0228] Cell lines were maintained in a humidified atmosphere at 37 °C containing 5% CO2.

[0229] With the exception of PC3 and SW480, all cell lines were routinely grown in minimal essential medium (DMEM) with phenol red (Sigma). PC3 and SW480 were grown in phenol red containing RPMI-1640 (Sigma).

[0230] Th...

Embodiment 3

[0244] In vitro antitumor activity of cannabis extracts against cancer cell lines

[0245] Purpose:

[0246] Robust methods and systems for high-throughput screening (HTS) to detect associations between cannabinoid ratios and doses and anti-tumor activity are provided. The method involves screening a growing library of human cancer cell lines tumor cell lines and / or biopsies using an expanded class range of cannabinoid-based compounds or extracts. The present invention demonstrates that testing of the biological activity of cannabis extracts, its components and compounds on different tumor cell lines or different tissue lines yields highly effective therapeutic data. In another aspect, the HTS systems and methods are applied to cell lines for the screening of potent cannabinoids or other natural extracts, with or without standard chemotherapy. Further, within the scope of the present application, HTS systems and methods are applied to patient-derived biopsies for screening e...

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PUM

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Abstract

The present invention discloses a method for high throughput screening (HTS) for identifying an analyte with a measurable effect on cells. The aforementioned method comprises steps of: (a) providing an array comprising a plurality of cell samples; (b) providing at least one analyte to be tested; (c) contacting said cell samples with said analyte; and (d) detecting a signal indicative of said measurable effect on cells wherein alteration of said signal over time measured on said cell sample relative to a control sample is indicative of said measurable effect of said analyte on said cell sample.The current invention further discloses means and methods for identifying an analyte selected from the group consisting of: cannabis extract or a fraction thereof cannabinoid- type constitute non cannabinoid-type constitute and any combination thereof. The analyte is indicative of cytotoxic or anti proliferative or anti mitotic or cell growth inhibitory activity in vitro.

Description

technical field [0001] The present invention relates to novel means and methods for high throughput screening of cancer cells. More specifically, the present invention relates to methods and systems for high throughput screening (HTS) for identifying analytes with measurable effects on cells. Background technique [0002] Cannabinoids include phytocannabinoids, endogenous endocannabinoids and synthetic cannabinoids. More than 60 phytocannabinoids have been identified within the Cannabis genus. The identification of cannabinoids exerts their pharmacological activity through two G-protein coupled receptors (GPCRs), the cannabinoid receptor type 1 (CB1) and receptor type 2 (CB2), in the cannabinoid signaling pathway. These receptors share 44% amino identity and unique but similar cannabinoid-binding profiles. CB1 receptors are mainly present in the central and peripheral nervous systems, inhibit neuronal excitability and mediator release, resulting in hypothermia, sedation, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/50
CPCG01N33/5014G01N33/5029G01N33/5085G01N33/948G01N33/5011A61P29/00A61P35/00A61P43/00A61K49/0008G01N33/502G01N33/5026
Inventor 埃亚勒·巴朗
Owner CANNABICS PHARMA
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