The Experts below are selected from a list of 660 Experts worldwide ranked by ideXlab platform
Wouter A Duetz - One of the best experts on this subject based on the ideXlab platform.
-
biocatalytic production of Perillyl Alcohol from limonene by using a novel mycobacterium sp cytochrome p450 alkane hydroxylase expressed in pseudomonas putida
Applied and Environmental Microbiology, 2005Co-Authors: J B Van Beilen, Bernard Witholt, Rene Holtackers, Daniel Luscher, Ulrich Bauer, Wouter A DuetzAbstract:A number of oxygenated monoterpenes present at low concentrations in plant oils have anticarcinogenic properties. One of the most promising compounds in this respect is ()-Perillyl Alcohol. Since this natural product is present only at low levels in a few plant oils, an alternative, synthetic source is desirable. Screening of 1,800 bacterial strains showed that many alkane degraders were able to specifically hydroxylate L-limonene in the 7 position to produce enantiopure ()-Perillyl Alcohol. The oxygenase responsible for this was purified from the best-performing wild-type strain, Mycobacterium sp. strain HXN-1500. By using N-terminal sequence information, a 6.2-kb ApaI fragment was cloned, which encoded a cytochrome P450, a ferredoxin, and a ferredoxin reductase. The three genes were successfully coexpressed in Pseudomonas putida by using the broad-host-range vector pCom8, and the recombinant converted limonene to Perillyl Alcohol with a specific activity of 3 U/g (dry weight) of cells. The construct was subsequently used in a 2-liter bioreactor to produce Perillyl Alcohol on a scale of several grams. The production of ()-Perillyl Alcohol from L-limonene is of interest because of the limited availability of ()-Perillyl Alcohol in nature and its proven anticarcinogenic properties; phase II trials to evaluate Perillyl Alcohol for the treatment of breast, pancreatic, and colorectal cancer are in progress (37). The only microbial enzyme system described thus far that transforms limonene to Perillyl Alcohol was found in Bacillus stearothermophilus BR388. However, this enzyme system is not sufficiently regiospecific; significant quantities of carveol, carvone, and terpineol are also produced (25). Removal of these side products is difficult as their boiling points and hydrophobicities are almost identical, and expensive purification methods (for example, chromatography) would be required to obtain sufficiently pure Perillyl Alcohol. Therefore, the industrial production of Perillyl Alcohol with this Bacillus enzyme system is not attractive. The conversion of limonene to perillic acid by a Pseudomonas putida strain expressing a cymene monooxygenase was described by Mars et al. (21) and could be interesting as Perillyl Alcohol is likely to be an intermediate in the production of perillic acid. Other literature concerning limonene biotransformations was reviewed recently (5). The approach that we used to find strains capable of regiospecific hydroxylation of limonene consisted of screening a collection of 1,800 bacterial strains grown on a range of relatively reduced substrates, such as toluene, naphthalene, and various alkanes. Using this approach, we anticipated that we would find oxygenases involved in catabolic pathways that would accept L-limonene as a substrate. Previous work has demonstrated that many catabolic oxygenases accept a wide range of unnatural substrates. Toluene dioxygenases, for ex
-
biotransformation of limonene by bacteria fungi yeasts and plants
Applied Microbiology and Biotechnology, 2003Co-Authors: Wouter A Duetz, Harro J. Bouwmeester, J B Van Beilen, Bernard WitholtAbstract:The past 5 years have seen significant progress in the field of limonene biotransformation, especially with regard to the regiospecificity of microbial biocatalysts. Whereas earlier only regiospecific biocatalysts for the 1,2 position (limonene-1,2-diol) and the 8-position (α-terpineol) were available, recent reports describe microbial biocatalysts specifically hydroxylating the 3-position (isopiperitenol), 6-position (carveol and carvone), and 7-position (Perillyl Alcohol, Perillylaaldehyde, and perillic acid). The present review also includes the considerable progress made in the characterization of plant P-450 limonene hydroxylases and the cloning of the encoding genes.
Pamela L Crowell - One of the best experts on this subject based on the ideXlab platform.
-
cell cycle arrest by the isoprenoids Perillyl Alcohol geraniol and farnesol is mediated by p21cip1 and p27kip1 in human pancreatic adenocarcinoma cells
Journal of Pharmacology and Experimental Therapeutics, 2007Co-Authors: Dean A Wiseman, Sean R Werner, Pamela L CrowellAbstract:Pancreatic cancer, the fourth leading cause of cancer-associated mortality in the United States, usually presents in an advanced stage and is generally refractory to chemotherapy. As such, there is a great need for novel therapies for this disease. The naturally derived isoprenoids Perillyl Alcohol, farnesol, and geraniol have chemotherapeutic potential in pancreatic and other tumor types. However, their mechanisms of action in these systems are not completely defined. In this study, we investigated isoprenoid effects on the cell cycle and observed a similar antiproliferative mechanism of action among the three compounds. First, when given in combination, the isoprenoids exhibited an additive antiproliferative effect against MIA PaCa-2 human pancreatic cancer cells. Furthermore, all three compounds induced a G(0)/G(1) cell cycle arrest that coincided with an increase in the expression of the cyclin kinase inhibitor proteins p21(Cip1) and p27(Kip1) and a reduction in cyclin A, cyclin B1, and cyclin-dependent kinase (Cdk) 2 protein levels. Immunoprecipitation studies demonstrated increased association of both p21(Cip1) and p27(Kip1) with Cdk2 as well as diminished Cdk2 kinase activity after isoprenoid exposure, indicating a cell cycle-inhibitory role for p21(Cip1) and p27(Kip1) in pancreatic adenocarcinoma cells. When siRNA was used to inhibit expression of p21(Cip1) and p27(Kip1) proteins in MIA PaCa-2 cells, conditional resistance to all three isoprenoid compounds was evident. Given similar findings in this cell line and in BxPC-3 human pancreatic adenocarcinoma cells, we conclude that the chemotherapeutic isoprenoid compounds Perillyl Alcohol, farnesol, and geraniol invoke a p21(Cip1)- and p27(Kip1)-dependent antiproliferative mechanism in human pancreatic adenocarcinoma cells.
-
effects of the isoprenoids Perillyl Alcohol and farnesol on apoptosis biomarkers in pancreatic cancer chemoprevention
Anticancer Research, 2002Co-Authors: Yvette D Burke, Siar A Ayoubi, Sean R Werner, Bryan C Mcfarland, Douglas K Heilman, Bruce A Ruggeri, Pamela L CrowellAbstract:Perillyl Alcohol, farnesol and geraniol have chemotherapeutic activity toward pancreatic and other cancers. Perillyl Alcohol induces apoptosis and increases expression of the proapoptotic protein Bak in cultured pancreatic tumor cells. We tested the hypothesis that farnesol and geraniol would have similar effects. After 48 hours of treatment with farnesol geraniol or Perillyl Alcohol, human BxPC3 pancreatic cancer cells exhibited a 3 to 10-fold increase in apoptosis and higher Bak expression than the controls. We then tested the hypotheses that Perillyl Alcohol and farnesol would have chemopreventive activity toward pancreatic cancer and would increase Bak expression and apoptosis in vivo. Hamster pancreatic cancer was initiated at time 0 with N-nitrosobis(2-oxopropyl)amine. Animals were fed control, 2% (w/w) Perillyl Alcohol, or 1% (w/w) farnesol diets from weeks 5-42. Pancreatic carcinoma incidence was decreased by Perillyl Alcohol and farnesol. Hyperplastic pancreatic ductal neoplasms from Perillyl Alcohol and farnesol-treated animals had higher Bak protein expression (p < 0.05), and somewhat higher apoptotic rates, diminished expression of the antiapoptotic protein BCL-XL, and lower rates of DNA synthesis than the controls.
-
induction of the apoptosis promoting protein bak by Perillyl Alcohol in pancreatic ductal adenocarcinoma relative to untransformed ductal epithelial cells
Carcinogenesis, 1997Co-Authors: K R Stayrook, J H Mckinzie, Y D Burke, Y A Burke, Pamela L CrowellAbstract:Perillyl Alcohol has antitumor activity toward pancreas and other cancers with low toxicity. Here, we have investigated the mechanism of action responsible for the differential sensitivity of malignant versus non-malignant pancreatic cells to the drug. We report that the rate of apoptosis is over 6-fold higher in Perillyl Alcohol-treated pancreatic adenocarcinoma cells than in untreated cells, and that the effect of Perillyl Alcohol on pancreatic tumor cells is significantly greater than its effect on non-malignant pancreatic ductal cells. Moreover, the Perillyl Alcohol-induced increase in apoptosis in all of the pancreatic tumor cells is associated with a 2- to 8-fold increase in the expression of the proapoptotic protein Bak, but Bak expression is not affected by Perillyl Alcohol in non-malignant cells. Thus, the antitumor activity of Perillyl Alcohol toward pancreatic cancers may be due to preferential stimulation of Bak-induced apoptosis in malignant versus normal cells. Bak may, therefore, be a useful biomarker for the chemopreventive and therapeutic effects of Perillyl Alcohol.
-
inhibition of pancreatic cancer growth by the dietary isoprenoids farnesol and geraniol
Lipids, 1997Co-Authors: Yvette D Burke, Jennifer M Stark, Steven L Roach, Stephanie E Sen, Pamela L CrowellAbstract:Fruits and vegetables have protective effects against many human cancers, including pancreatic cancer. Isoprenoids are one class of phytochemicals which have antitumor activity, but little is known about their effects on cancer of the pancreas. We tested the hypothesis that isoprenoids would inhibit the growth of pancreatic tumor cells. Significant (60-90%) inhibition of the anchorage-independent growth of human MIA PaCa2 pancreatic tumor cells was attained with 25 microM farnesol, 25 microM geranylgeraniol, 100 microM Perillyl amine, 100 microM geraniol, or 300 microM Perillyl Alcohol. We then tested the relative in vivo antitumor activities of dietary farnesol, geraniol, and Perillyl Alcohol against transplanted PC-1 hamster pancreatic adenocarcinomas. Syrian Golden hamsters fed geraniol or farnesol at 20 g/kg diet exhibited complete inhibition of PC-1 pancreatic tumor growth. Both farnesol and geraniol were more potent than Perillyl Alcohol, which inhibited tumor growth by 50% at 40 g/kg diet. Neither body weights nor plasma cholesterol levels of animals consuming isoprenoid diets were significantly different from those of pair-fed controls. Thus, farnesol, geraniol, and Perillyl Alcohol suppress pancreatic tumor growth without significantly affecting blood cholesterol levels. These dietary isoprenoids warrant further investigation for pancreatic cancer prevention and treatment.
J B Van Beilen - One of the best experts on this subject based on the ideXlab platform.
-
biocatalytic production of Perillyl Alcohol from limonene by using a novel mycobacterium sp cytochrome p450 alkane hydroxylase expressed in pseudomonas putida
Applied and Environmental Microbiology, 2005Co-Authors: J B Van Beilen, Bernard Witholt, Rene Holtackers, Daniel Luscher, Ulrich Bauer, Wouter A DuetzAbstract:A number of oxygenated monoterpenes present at low concentrations in plant oils have anticarcinogenic properties. One of the most promising compounds in this respect is ()-Perillyl Alcohol. Since this natural product is present only at low levels in a few plant oils, an alternative, synthetic source is desirable. Screening of 1,800 bacterial strains showed that many alkane degraders were able to specifically hydroxylate L-limonene in the 7 position to produce enantiopure ()-Perillyl Alcohol. The oxygenase responsible for this was purified from the best-performing wild-type strain, Mycobacterium sp. strain HXN-1500. By using N-terminal sequence information, a 6.2-kb ApaI fragment was cloned, which encoded a cytochrome P450, a ferredoxin, and a ferredoxin reductase. The three genes were successfully coexpressed in Pseudomonas putida by using the broad-host-range vector pCom8, and the recombinant converted limonene to Perillyl Alcohol with a specific activity of 3 U/g (dry weight) of cells. The construct was subsequently used in a 2-liter bioreactor to produce Perillyl Alcohol on a scale of several grams. The production of ()-Perillyl Alcohol from L-limonene is of interest because of the limited availability of ()-Perillyl Alcohol in nature and its proven anticarcinogenic properties; phase II trials to evaluate Perillyl Alcohol for the treatment of breast, pancreatic, and colorectal cancer are in progress (37). The only microbial enzyme system described thus far that transforms limonene to Perillyl Alcohol was found in Bacillus stearothermophilus BR388. However, this enzyme system is not sufficiently regiospecific; significant quantities of carveol, carvone, and terpineol are also produced (25). Removal of these side products is difficult as their boiling points and hydrophobicities are almost identical, and expensive purification methods (for example, chromatography) would be required to obtain sufficiently pure Perillyl Alcohol. Therefore, the industrial production of Perillyl Alcohol with this Bacillus enzyme system is not attractive. The conversion of limonene to perillic acid by a Pseudomonas putida strain expressing a cymene monooxygenase was described by Mars et al. (21) and could be interesting as Perillyl Alcohol is likely to be an intermediate in the production of perillic acid. Other literature concerning limonene biotransformations was reviewed recently (5). The approach that we used to find strains capable of regiospecific hydroxylation of limonene consisted of screening a collection of 1,800 bacterial strains grown on a range of relatively reduced substrates, such as toluene, naphthalene, and various alkanes. Using this approach, we anticipated that we would find oxygenases involved in catabolic pathways that would accept L-limonene as a substrate. Previous work has demonstrated that many catabolic oxygenases accept a wide range of unnatural substrates. Toluene dioxygenases, for ex
-
biotransformation of limonene by bacteria fungi yeasts and plants
Applied Microbiology and Biotechnology, 2003Co-Authors: Wouter A Duetz, Harro J. Bouwmeester, J B Van Beilen, Bernard WitholtAbstract:The past 5 years have seen significant progress in the field of limonene biotransformation, especially with regard to the regiospecificity of microbial biocatalysts. Whereas earlier only regiospecific biocatalysts for the 1,2 position (limonene-1,2-diol) and the 8-position (α-terpineol) were available, recent reports describe microbial biocatalysts specifically hydroxylating the 3-position (isopiperitenol), 6-position (carveol and carvone), and 7-position (Perillyl Alcohol, Perillylaaldehyde, and perillic acid). The present review also includes the considerable progress made in the characterization of plant P-450 limonene hydroxylases and the cloning of the encoding genes.
Thomas C. Chen - One of the best experts on this subject based on the ideXlab platform.
-
Potentially Curative Therapeutic Activity of NEO212, a Perillyl Alcohol-Temozolomide Conjugate, in Preclinical Cytarabine-Resistant Models of Acute Myeloid Leukemia
'MDPI AG', 2021Co-Authors: Axel H. Schönthal, Heeyeon Cho, Steve Swenson, Radu O. Minea, Hye Na Kim, Nazleen Mohseni, Yong-mi Kim, Thomas C. ChenAbstract:Despite progress in the treatment of acute myeloid leukemia (AML), the clinical outcome remains suboptimal and many patients are still dying from this disease. First-line treatment consists of chemotherapy, which typically includes cytarabine (AraC), either alone or in combination with anthracyclines, but drug resistance can develop and significantly worsen prognosis. Better treatments are needed. We are developing a novel anticancer compound, NEO212, that was created by covalent conjugation of two different molecules with already established anticancer activity, the alkylating agent temozolomide (TMZ) and the natural monoterpene Perillyl Alcohol (POH). We investigated the anticancer activity of NEO212 in several in vitro and in vivo models of AML. Human HL60 and U937 AML cell lines, as well as different AraC-resistant AML cell lines, were treated with NEO212 and effects on cell proliferation, cell cycle, and cell death were investigated. Mice with implanted AraC-sensitive or AraC-resistant AML cells were dosed with oral NEO212, and animal survival was monitored. Our in vitro experiments show that treatment of cells with NEO212 results in growth inhibition via potent G2 arrest, which is followed by apoptotic cell death. Intriguingly, NEO212 was equally potent in highly AraC-resistant cells. In vivo, NEO212 treatment strikingly extended survival of AML mice and the majority of treated mice continued to thrive and survive without any signs of illness. At the same time, we were unable to detect toxic side effects of NEO212 treatment. All in all, the absence of side effects, combined with striking therapeutic activity even in an AraC-resistant context, suggests that NEO212 should be developed further toward clinical testing
-
the rolipram Perillyl Alcohol conjugate neo214 is a mediator of cell death through the death receptor pathway
Molecular Cancer Therapeutics, 2019Co-Authors: Heeyeon Cho, Axel H. Schönthal, Florence M. Hofman, Weijun Wang, Thu Zan Thein, Stephen Swenson, Rochelle A Fayngor, Nagore I Marinramos, Thomas C. ChenAbstract:Glioblastoma (GBM) is a highly aggressive primary brain tumor with a poor prognosis. Treatment with temozolomide, standard of care for gliomas, usually results in drug resistance and tumor recurrence. Therefore, there is a great need for drugs that target GBM. NEO214 was generated by covalently linking rolipram to Perillyl Alcohol (POH) via a carbamate bond to form the rolipram–Perillyl Alcohol conjugate. We show here that NEO214 is effective against both temozolomide-sensitive and temozolomide-resistant glioma cells. Furthermore, NEO214 is effective for different mechanisms of temozolomide resistance: overexpression of MGMT (O6-methylguanine methyl-transferase); deficiency in specific mismatch repair proteins; and overexpression of base excision repair (BER) proteins. NEO214-induced cytotoxicity involves apoptosis triggered by endoplasmic reticulum (ER) stress, as well as activating the Death Receptor 5 (DR5)/TNF-related apoptosis-inducing ligand (TRAIL/Apo2L) pathway. In vitro studies show that glioma cells treated with NEO214 express DR5 and exhibit cell death in the presence of recombinant TRAIL, a growth factor constitutively produced by astrocytes. Our in vitro 3D coculture data show that induction of DR5 in glioma cells with NEO214 and TRAIL cause tumor cell death very effectively and specifically for glioma cells. In vivo studies show that NEO214 has antitumor efficacy in orthotropic syngeneic rodent tumor models. Furthermore, NEO214 has therapeutic potential especially for brain tumors because this drug can cross the blood–brain barrier (BBB), and is effective in the TRAIL-rich astrocyte microenvironment. NEO214 is a strong candidate for use in the treatment of GBMs.
-
induction of pro apoptotic endoplasmic reticulum stress in multiple myeloma cells by neo214 Perillyl Alcohol conjugated to rolipram
International Journal of Molecular Sciences, 2018Co-Authors: Thomas C. Chen, Florence M. Hofman, Heeyeon Cho, Nymph Chan, Shirin Labib, Axel H. SchönthalAbstract:Despite the introduction of new therapies for multiple myeloma (MM), many patients are still dying from this disease and novel treatments are urgently needed. We have designed a novel hybrid molecule, called NEO214, that was generated by covalent conjugation of the natural monoterpene Perillyl Alcohol (POH), an inducer of endoplasmic reticulum (ER) stress, to rolipram (Rp), an inhibitor of phosphodiesterase-4 (PDE4). Its potential anticancer effects were investigated in a panel of MM cell lines. We found that NEO214 effectively killed MM cells in vitro with a potency that was over an order of magnitude stronger than that of its individual components, either alone or in combination. The cytotoxic mechanism of NEO214 involved severe ER stress and prolonged induction of CCAAT/enhancer-binding protein homologous protein (CHOP), a key pro-apoptotic component of the ER stress response. These effects were prevented by salubrinal, a pharmacologic inhibitor of ER stress, and by CHOP gene knockout. Conversely, combination of NEO214 with bortezomib, a drug in clinical use for patients with MM, resulted in synergistic enhancement of MM cell death. Combination with the adenylate cyclase stimulant forskolin did not enhance NEO214 impact, indicating that cyclic adenosine 3',5'-monophosphate (AMP) pathways might play a lesser role. Our study introduces the novel agent NEO214 as a potent inducer of ER stress with significant anti-MM activity in vitro. It should be further investigated as a potential MM therapy aimed at exploiting this tumor's distinct sensitivity to ER stress.
-
Temozolomide-Perillyl Alcohol conjugate impairs Mitophagy flux by inducing lysosomal dysfunction in non-small cell lung Cancer cells and sensitizes them to irradiation
'Springer Science and Business Media LLC', 2018Co-Authors: Minghui Chang, Thomas C. Chen, Weijun Wang, Xingguo Song, Xinran Geng, Xingwu Wang, Li Xie, Xianrang SongAbstract:Abstract Background Temozolomide-Perillyl Alcohol conjugate (TMZ-POH), a novel Temozolomide (TMZ) analog developed based on the conjugation of TMZ and Perillyl Alcohol (POH), displayed strong anticancer potency in multiple cancer types. In this study, we aimed to clarify the relationship between TMZ-POH and autophagy, and explore the underlying mechanisms involved in. Methods The proteins involved in autophagy, mitochondrial fission, lysosomal function and membrane traffic were detected by western blots; Autophagosome, mitochondria and lysosome were visualized by transmission electron microscope (TEM) and immunostaining; Apoptosis analysis and fluorescence probe detection were applied by flow cytometry. Results TMZ-POH blocked mitophagy flux although the number of autophagosomes which colocalized with mitochondria in the cells was increased via inducing lysosomal dysfunction as evidence from impaired lysosomal acidification, maturation and hampered autophagosome- lysosome fusion, which largely depended on its downregulation on the small GTPase RAB7A via mevalonate pathway. More importantly, our data demonstrated TMZ-POH sensitized cancer cell to irradiation induced apoptosis. Conclusions Temozolomide-Perillyl Alcohol conjugate impairs mitophagy flux by inducing lysosomal dysfunction in Non-Small Cell Lung Cancer (NSCLC) cells and sensitizes them to irradiation, thereby proposing TMZ-POH as a potential radiosensitizer
-
Intranasal Perillyl Alcohol for Glioma Therapy: Molecular Mechanisms and Clinical Development
'MDPI AG', 2018Co-Authors: Thomas C. Chen, Clovis O. Da Fonseca, Axel H. SchönthalAbstract:Intracranial malignancies, such as primary brain cancers and brain-localized metastases derived from peripheral cancers, are particularly difficult to treat with therapeutic agents, because the blood-brain barrier (BBB) effectively minimizes brain entry of the vast majority of agents arriving from the systemic circulation. Intranasal administration of cancer drugs has the potential to reach the brain via direct nose-to-brain transport, thereby circumventing the obstacle posed by the BBB. However, in the field of cancer therapy, there is a paucity of studies reporting positive results with this type of approach. A remarkable exception is the natural compound Perillyl Alcohol (POH). Its potent anticancer activity was convincingly established in preclinical studies, but it nonetheless failed in subsequent clinical trials, where it was given orally and displayed hard-to-tolerate gastrointestinal side effects. Intriguingly, when switched to intranasal delivery, POH yielded highly promising activity in recurrent glioma patients and was well tolerated. As of 2018, POH is the only intranasally delivered compound in the field of cancer therapy (outside of cancer pain) that has advanced to active clinical trials. In the following, we will introduce this compound, summarize its molecular mechanisms of action, and present the latest data on its clinical evaluation as an intranasally administered agent for glioma
Yumi Fujiwara - One of the best experts on this subject based on the ideXlab platform.
-
molecular cloning and characterization of a perilla frutescens cytochrome p450 enzyme that catalyzes the later steps of perillaldehyde biosynthesis
Phytochemistry, 2017Co-Authors: Yumi Fujiwara, Michiho ItoAbstract:Perilla produces the cyclohexanoid monoterpene perillaldehyde as a major constituent of an essential oil that is accumulated in its glandular trichomes. Perillaldehyde is a marker compound for quality control of soyo and has biological activities such as antibacterial, sedative, or vasodilatory effects. The predicted perillaldehyde formation involves the cyclization of geranyl diphosphate, hydroxylation, and oxidation, and cytochrome P450 plays a crucial role in perillaldehyde biosynthesis. In this study, a cytochrome P450-type enzyme with Perillyl Alcohol and perillaldehyde synthase activities was isolated by analyzing an expressed sequence tag library from several oil types of pure lines of perilla. A recombinant protein with a sequence that was highly specific for the type of perillaldehyde was expressed in Saccharomyces cerevisiae and evaluated by an in vitro enzymatic reaction. The recombinant protein catalyzed the hydroxylation and oxidation of limonene to Perillyl Alcohol and perillaldehyde. Cytochrome P450 limonene-7-hydroxylase cDNA from Perilla frutescens has been previously isolated. The cytochrome P450 isolated in this study shares 37% amino-acid identity with the previously isolated enzyme; however, it may have different characteristics.