A composition of limonene and ibuprofen, in the appropriate molar ratio to form a deep eutectic system.
This invention is a composition of limonene and ibuprofen, in the appropriate molar ratio to form a deep eutectic system. These systems have proven to behave as a single entity, distinct from that of the pure components that constitute the mixture. Limonene:ibuprofen eutectic systems present protective, anti-inflammatory and anti-cancer properties and have demonstrated to enhance its therapeutic action, when compared to the pure components. The invention herein proposed provides clues for future development of new pharmaceutical systems, based on eutectic compositions of terpenes and anti-inflammatory agents which may enhance the bioactivity and efficacy of the systems designed and can lead to significant breakthroughs in cancer therapy.
Carcinogenesis is a phenomenon not only restricted to the abnormal growth of cells but also includes angiogenesis and inflammation processes, which play an important role in tumor progression. The bioactivity of deep eutectic solvents (DES) is not yet well explored. Literature has already reported cytotoxicity of ammonium- and choline chloride-based DES for several cancer cell lines, but it is not clear if this is a selective toxicity towards cancer cell lines.
Limonene is a cyclic monoterpene studied at preclinical and clinical levels due to its chemopreventive and chemotherapeutic activities at several types of cancer, as lung, breast, gastric or prostate. Its high lipophilicity contributes to a favorable cellular absorption, specifically at intestinal level, leading to a good bioavailability in the systemic circulation. In colorectal cancer, limonene has been reported to induce apoptosis via mitochondrial pathway and affect PI3K/Akt signaling pathway (survival and apoptosis). However, the administration of limonene is limited by its toxicity and volatility.
On the other hand, regarding inflammation process, different molecules have been explored. For example, fatty acids terpenes and non-steroidal anti-inflammatory agents have already been reported, where (i) capric acid, is known to act against oxidative stress and pro-inflammatory cytokines, (ii) menthol is capable of reducing IL-1ß at chronic colonic inflammation, and (iii) ibuprofen is a commercialized anti-inflammatory compound also associated with tumor reduction.
The problems associated with the use of limonene as a potent anti-cancer agent are related to its toxicity and volatility. To overcome this, different therapeutic deep eutectic solvents (THEDES) based on limonene (LIM) were developed to unravel the anticancer potential of such systems. THEDES were prepared by gently mixing capric acid (CA), menthol or ibuprofen (IBU) with LIM. Successful eutectic mixtures were obtained for Menthol:LIM (1:1), CA:LIM (1:1), IBU:LIM (1:4) and IBU:LIM(1:8).
LIM-based THEDES were evaluated in terms of their cytotoxicity and antiproliferative effects. The results indicate that all the THEDES present antiproliferative properties, but IBU:LIM (1:4) was the only formulation able to inhibit HT29 proliferation without comprising cell viability. Therefore, IBU:LIM (1:4) was the formulation selected for further assessment of anticancer properties. The results suggest that the mechanism of action of LIM:IBU (1:4) is different from isolated IBU and LIM. The combination of limonene with ibuprofen decreases the toxicity of limonene towards healthy cells by a factor of 4. This benefit could potentiate the use of limonene as an anti-cancer therapeutic agent. Thus, THEDES comprises the protective and anti-inflammatory properties of IBU allied to the anticancer properties of LIM.
Unpublished details and trade secrets of the project, including new data, is undisclosed to date but researchers would be more than happy to discuss with interested parties under an NDA (this can be easily arranged via the University).
STAGE OF DEVELOPMENT
TRL 3 – analytical and experimental critical function and/or characteristic proof of concept.
- Enhanced solubility of ibuprofen: 4-fold increase;
- THEDES (Limonene:ibuprofen): 4.4 times more selective towards cancer cell lines than limonene alone;
- THEDES (Limonene:ibuprofen): 6x less toxic towards healthy cells than limonene + ibuprofen (physical mixture);
- Protective and anti-inflammatory activity: Increase the inhibition of production of reactive oxide species by 1.4x;
- Anti-cancer activity.
- Cancer therapeutics.
- Available for exclusive and non-exclusive licensing;
- Seeking co-development partners to extend the research on other possible combinations, design and prepare in vitro testing for the understanding of the mechanisms of action of the system.
- Patent application submitted (WO2020188547).
Carolina Velhinho Pereira
Liliana Andreia Amaro Rodrigues
Ana Rita Duarte