Use of cannabidiol in the treatment of tuberous sclerosis complex

Inactive Publication Date: 2020-07-02
GW RES LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0029]In accordance with a second aspect of the present invention there is provided a method of treating a patient with Tuberous Sclerosis Complex (TSC) comp

Problems solved by technology

When present, neurologic complications are the most common cause of mortality and morbidity and are the most likely to affect quality of life.
If the tumours are large or th

Method used

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  • Use of cannabidiol in the treatment of tuberous sclerosis complex
  • Use of cannabidiol in the treatment of tuberous sclerosis complex
  • Use of cannabidiol in the treatment of tuberous sclerosis complex

Examples

Experimental program
Comparison scheme
Effect test

example 1

Efficacy of Cannabidiol in Reduction of Tumours in Two Tuberous Sclerosis Complex Cell Lines

Materials and Methods

Cell Lines and Maintenance

[0047](i) Angiomyolipomas (AML) cells derived from a Tuberous Sclerosis Complex patient (621-101 cell-line), with a known loss of function of TSC2. To generate a rescue control cell line, these 621-101 cells were stably transfected with a TSC2-expressing plasmid possessing Zeocin™ resistance (pcDNA3.1zeo-hTSC2). Zeocin™ was purchased from LifeTechnologies (cat no: R25001) and was used to originally generate the stable cell line (by Prof. Lisa Henske) over two weeks of Zeocin™ selection and then for continued maintenance of the cells in tissue culture (at 100 μg / ml).

[0048](ii) Tsc2− / − p53− / − and Tsc2+ / + p53− / − mouse embryonic fibroblast (MEF) cell lines (now referred to as Tsc2− / − and Tsc2+ / +, respectively).

Cell Culture

[0049]Tsc2− / − and Tsc2+ / + cell lines were cultured and maintained in DMEM supplemented with 10% (v / v) FBS and 1% (v / v) penicillin-...

example 2

Efficacy of Cannabidiol in Reduction of Tumours a Zebrafish Model of Tuberous Sclerosis Complex

Materials and Methods

Zebrafish Husbandry

[0071]A zebrafish model of TSC with a tsc2 gene deletion was used. This model has been previously published and validated (Kim et al. 2011). By mating the gene deletion animals with wild-type tsc2+ / + animals, heterozygote (tsc2+ / −) zebrafish were also obtained and tested in this example.

Imaging

[0072]For both pS6 and TUNEL staining, non-consecutive sections were imaged using a Zeiss AxioImager microscope. Exposure time was kept constant during image acquisition and determined by the observation of a slide where primary antibody or enzyme solution was omitted. Pictures were taken with a 20× objective, using the AxioVision software, and coloured using Fiji ImageJ. All counting and measuring of cells was done in the original black and white pictures.

Immunohistochemistry

[0073]For TUNEL staining (1:10; Roche, Sigma, UK), 10 μm sections were cut and collect...

example 3

Efficacy of Cannabidiol in Combination With an mTOR Inhibitor in a Tumour Cell Line

Materials and Methods

Cell Lines

[0096]PANC-1 is a human pancreatic carcinoma, epithelial-like cell line; PANC-1 cells are used as an in vitro model of non-endocrine pancreatic cancer for tumorigenicity studies. The cells possess the type B phenotype for glucose-6-phosphate dehydrogenase G6PD and overexpress heregulin / human epidermal growth-factor receptor 2 (HER2 / neu) oncogene.

[0097]MIAPACA is a homo sapiens pancreas carcinoma. This is a hypotriploid human cell line.

Materials

[0098]Everolimus (RAD001, Afinitor, Novartis), is a rapamycin analogue. It is an oral mammalian target of rapamycin (mTOR) inhibitor and belongs to the PI3K related family of protein kinases and is activated by phosphorylation at serine 2448 (S2448).

[0099]Pure CBD was used in this example.

Cell Viability Assay

[0100]Cell viability was analyzed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay. Pancreati...

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Abstract

The present invention relates to the use of cannabidiol (CBD) for the treatment of tumours associated with Tuberous Sclerosis Complex (TSC). In particular the CBD was able to decrease the number and size of marker cells, pS6, in a zebrafish model of TSC. This is5 suggestive of a disease modifying effect whereby treatment with CBD could result in the reduction or prevention of the benign tumours that occur in TSC patients. Preferably the CBD used is in the form of a highly purified extract of cannabis such that the CBD is present at greater than 98% of the total extract (w/w) and the other components of the extract are characterised. In particular the cannabinoid tetrahydrocannabinol (THC) has been substantially 10 removed, to a level of not more than 0.15% (w/w) and the propyl analogue of CBD, cannabidivarin, (CBDV) is present in amounts of up to 1%. Alternatively, the CBD may be a synthetically produced CBD. In use the CBD is given concomitantly with one or more other drugs used in the treatment of TSC. Such drugs may include rapamycin and/or everolimus.

Description

[0001]The present invention relates to the use of cannabidiol (CBD) for the treatment of tumours associated with Tuberous Sclerosis Complex (TSC). In particular the CBD was able to decrease the number and size of marker cells, pS6, in a zebrafish model of TSC. This is suggestive of a disease modifying effect whereby treatment with CBD could result in the reduction or prevention of the benign tumours that occur in TSC patients.[0002]Preferably the CBD used is in the form of a highly purified extract of cannabis such that the CBD is present at greater than 98% of the total extract (w / w) and the other components of the extract are characterised. In particular the cannabinoid tetrahydrocannabinol (THC) has been substantially removed, to a level of not more than 0.15% (w / w) and the propyl analogue of CBD, cannabidivarin, (CBDV) is present in amounts of up to 1%. Alternatively, the CBD may be a synthetically produced CBD.[0003]In use the CBD is given concomitantly with one or more other d...

Claims

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Application Information

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IPC IPC(8): A61K31/05A61P35/00
CPCA61K45/06A61P35/00A61K31/05A61K31/436A61K31/352A61K2300/00A61P43/00
Inventor WHALLEY, BENJAMINHIND, WILLIAMGRAY, ROYSTONBAZELOT, MICHAELDE SILVA SERRA, INESWILLIAMS, CLAIRETEE, ANDREW
Owner GW RES LTD
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