Synthesis of (-)-Camphene derivatives and evaluation of their biological activities

Terpenes are aromatic oils that give their fragrance and taste. These compounds have additionally been acknowledged to hold an array of medicinal advantages, (±)-camphene is a one of example cannabis terpene possessing a strong musky and earthy smell with hints of pine. (±)-Camphene is a primary ingredient in many perfumes, oils, and topical lotions. It is also naturally present in conifers, nutmeg, ginger, and rosemary. A common synthetic method for (±)-camphene is to use pinene, another cannabis terpene. (±)-Camphene is commonly used in some of salves, creams and lotions that are cannabis-based and its possibility of this ability to treat skin conditions such as eczema and psoriasis indicates camphene could hold antifungal and antibacterial effectiveness. Camphene is generally more commonly found in indica strains with medicinal potential and is notable. In this study, (-)-camphene (70) will be used as key compounds for the synthesis of new biologically active compounds (73-77). Starting from (-)-camphene [75% (camphene with 90% fenchene)], bromination reactions and Suzuki-Miyaura coupling reactions will be carried out. In this bromination system, (-)-camphene (70) will be dissolved in dichloromethane (20 mL), and Br2 (dissolved in CH2Cl2) will be added dropwise to the reaction mixture with the help of a pressure balanced dropping funnel. When aniline is added to dibromo-(-)-camphene (71) and the reaction mixture is boiled under reflux for 2 hours, (1S,4R)-3-(Bromomethylene)-2,2-dimethylbicyclo-2,2-dimetilbisiklo[2.2.1]heptane (72) will be obtained. Brominated derivatives suitable for metal-catalyzed reactions are an important key compound for many new products. In this study, it is planned to apply metal-catalyzed Suzuki-Miyaura coupling reactions for the synthesis of new (-)-camphene derivatives from monobromo-(-)-camphene. Coupling reactions of bicyclic structures were carried out in the presence of catalysts such as Pd(PPh3)4 and various boronic acids (phenyl, 4-methoxyphenyl, 4-thiomethylphenyl, 4-ethylphenyl, 4-trifluoromethoxyphenyl). Following the general synthesis procedure for the synthesis of relevant compounds, monobromo-(-)-camphene (72) and related boronic acid compounds (73-77) were interacted with, then K2CO3 was added to the reaction mixture as a base, toluene ethanol and water as solvents (1:1:1) was added and boiled under reflux at 110°C for 17 hours. Then, the crude product was extracted with ethylacetate (3x50 mL) and the organic phase was dried with Na2SO4 and the reaction mixture was purified by silica gel column chromatography. The final compounds were characterized using 1H NMR, 13C NMR and FTIR spectroscopy methods; they were also shown to be photoactive and negatively polarimetric. The biological effects of (-)-camphene derivatives (cancer cell lines and cell culture, cytotoxicity test, DNA fragmentation by agarose gel electrophoresis, migration assay, antibacterial and antifungal activity measurement by microdilution assay) were examined. In this study, A549 (ATCC, CCL-185), Calu1 (ATCC, HTB-54), and H1650 (ATCC, CRL-5883) lung cancer cell lines, SW1353 (ATCC, HTB-94)/MG63 (ATCC, CRL-1427)/, and Saos2 (ATCC, HTB-85) bone cancer cell lines], Beas2B (RRID, CVCL-0168) normal lung cell line, and HC (Sigma Aldrich, 402-05A) normal chondrocytes cell line were used. MTT test was used to measure the effects of the synthesized compounds (71-77) on cell proliferation and NCI-60 survival parameter values. It was determined by the LDH method whether the compounds to be tested (71-77) were cell cytotoxic or cytostatic. DNA laddering activity of the (-)-camphene compounds was evaluated by a DNA laddering assay in accordance with the standard method. Wound healing assay was used to determine the effect of the compounds on the migration of cells. It is a method based on measuring the proliferation ability of cells in the medium containing the test substance. For measuring the antibacterial and antifungal activities using microdilution test in this project [E. faecalis VRE ATCC 19433, E. faecalis ATCC 29212, S. aureus MSSA ATCC 29213, S. aureus ATCC 25923, S. aureus MRSA ATCC 46300, E. coli ATCC 25922, P. aeruginosa AGME ATCC 27853, C. albicans (ATCC 10231), S. gordonii (NCTC 7870) and A. actinomycetemcomitans (ATCC 33384)] were used. Therefore, these molecules lack antimicrobial properties against all bacteria and fungi but they have the effect of (-)-camphene molecules are partly responsible for their anticancer activities it may be a good option for cancer treatment and compound 72 was more cancer-specific for bone and lung cancer types, and make it is the most convenient one for further preclinical studies.