Published Research
Published research on Ubisol-Aqua™ describes the discovery, solubilization capabilities, and preclinical studies in animal and cell models that demonstrate the efficacy of this new technology for solubilizing lipophillic compounds for potential use in various applications. The Ubisol-Aqua™ technology has been tested with two water-insoluble model compounds, coenzyme Q10 (CoQ10) and amphotericin B, to demonstrate its broad applicability. These studies demonstrate that Ubisol-Aqua™ can be used to:
1. Enhance solubility of water-insoluble compounds
2. Improve bioavailability
3. Reformulate drugs to reduce toxicity
4. Act as a pro-drug
Zymes has also begun making significant contributions to research in the area of green chemistry. Zymes’ proprietary PTS, a nanomicelle-forming amphiphile, can be used to promote chemical reactions in water at room temperature by cost-effectively achieving solubilization of both reactants and catalysts. This can obviate the need for organic solvents and heat, both of which are potentially either costly, or detrimental to the environment. This approach highlights micellar catalysis, a reaction variable that has been relatively underdeveloped in the synthetic community, particularly among those working in transition metal based cross-couplings. These studies demonstrate the use of PTS as a new enabling technology in micellar catalysis for transition metal-mediated processes.
Published Papers:
Borowy-Borowski H, Sodia C, Docherty J, Walker PR, Sikorska M. Unique technology for solubilization and delivery of highly lipophilic bioactive molecules. Journal of Drug Targeting 2003;12(7):415-424.
The Ubisol-Aqua™ family of compounds was expanded to produce an assortment of novel carriers capable of solubilizing water-insoluble compounds. The new carriers included polyoxyethanyl-cholesteryl sebacate (PCS™) and polyoxyethanyl-β-sitosteryl sebacate (PSS™). The compounds were synthesized, purified and evaluated for safety. In vivo and in vitro safety studies demonstrated that the compounds were non-toxic and efficacious. Acute cardiotoxicity studies performed in rats further demonstrated the safety of the compounds. PTS™, PCS™, and PSS™ formed stable, water-soluble nano-sized complexes with a variety of water-insoluble molecules including vitamins, carotenoids, ubiquinones, poly-unsaturated fatty acids, and polyene macrolide antibiotics.
In vivo bioavailability experiments with PCS™: Coenzyme Q10 showed increased blood levels of coenzyme Q10 relative to control rats given an oil-based coenzyme Q10 formulation. In addition, when rats were given PTS™ alone, plasma vitamin E levels increased, demonstrating the ability of PTS™ to act as a pro-drug. When amphotericin B was solubilized with PSS™, it was effective against a systemic c. neoformans infection in mice and was as effective as the antifungal drug, FUNGIZONE. In vitro toxicity studies showed it to be similar to AMBISOME, a commercially available liposomal preparation of amphotericin B. These exciting new formulations represent a major advance in the delivery of water-insoluble molecules with wide application for delivery of ingredients for the nutritional, cosmeceutical and pharmaceutical industries. These studies further demonstrate the vast potential of the technology including the reformulation of drugs with near term patent expiry or those that have failed clinical trials due to low solubility.
Sikorska M, Borowy-Borowski H, Zurakowski B, Walker PR. Derivatised alpha-tocopherol as a CoQ10 carrier in a novel water-soluble formulation. Biofactors 2003;18(1-4):173-83.
This study describes the synthesis of polyoxyethanyl-α-tocopheryl sebacate (PTS™) and its use to successfully solubilize coenzyme Q10. PTS™ and coenzyme Q10 form a non-covalent complex (PTS™: Coenzyme Q10) that is water-soluble and stable in solution for extended periods of time. Following oral administration of this formulation to rats, blood levels of both vitamin E and coenzyme Q10 were measured. Blood levels of coenzyme Q10 were elevated within 1 hour, peaked at 3 hours and remained elevated at 20 hours post administration. Vitamin E concentrations were 3 to 4 times higher than baseline levels observed 3-9 hours after ingestion and remained elevated at 24 hours. These results demonstrate that the active ingredients in PTS™ are absorbed rapidly through the gastrointestinal tract and are bioavailable.
The newly formulated, water-soluble coenzyme Q10 was also tested in a rat model of ishemic brain damage induced by disruption of blood flow to the brain. Rats received PTS: Coenzyme Q10 by intraperitoneal injections given immediately and 3 hours after ischemia was induced. Brain damage was assessed by histological evaluation. Rats treated with PTS™: Coenzyme Q10 experienced significant neuroprotection relative to untreated rats. This study demonstrates the potential therapeutic value of water-soluble coenzyme Q10 in treating acute ischemic conditions such as stroke and heart attack.
Sandhu JK, Pandey S, Ribecco-Lutkiewicz M, Monette R, Borowy-Borowski H, Walker PR, Sikorska M. Molecular mechanisms of glutamate neurotoxicity in mixed cultures of NT2 derived neurons and astrocytes: protective effects of Coenzyme Q10. J. of Neuroscience Research 2003;72:691-703.
In the central nervous system, glutamate acts as a neurotransmitter but, at high concentrations, can also act as a neurotoxin through a process called excitotoxicity. Excitotoxicity contributes to nerve damage in neurologic disorders such as stroke, epilepsy, hypoglycemia, Alzheimer’s, Parkinson’s, Huntington’s disease and amyotrophic lateral sclerosis (ALS). Excitotoxicity causes nerve damage through the formation of reactive nitrogen and reactive oxygen species (RNOS, ROS), which cause neuronal cell death by decreasing energy (ATP) production in the mitochondria. Astrocytes are non-nerve cells in the brain which help to rapidly clear glutamate, preventing excitotoxic injury to nerve cells. In an experimental model where nerve cells are exposed to glutamate and low oxygen, astrocytes will no longer clear glutamate and accordingly, nerve cell death occurs. However, a number of antioxidants including coenzyme Q10 and vitamin E are able to scavenge RNOS and protect nerve cells from the damaging effects induced by glutamate.
This study demonstrated that pretreatment of cultured nerve cells with Ubisol-Aqua™: Coenzyme Q10 increased mitochondrial coenzyme Q10 and ATP levels. Ubisol-Aqua™: Coenzyme Q10 also effectively decreased RNOS production, decreased indicators of cell damage and death, and improved astrocyte function, significantly increasing nerve cell survival.
Chuang Y, Chan JYH, Chang, AYW, Sikorska M, Borowy-Borowski H, Liou C, Cha SHH, Neuroprotective effects of coenzyme Q10 at rostral ventrolateral medulla against fatality during experimental endotoxemia in the rat. Shock 2003;19(5):427-432.
Sepsis is a systemic inflammatory response to infection or injury which continues to have high morbidity and mortality despite aggressive treatment. In this animal study, sepsis-induced death was usually preceded by a dramatic reduction in the low frequency components of blood pressure signals. The presence of these signals reflects the functional integrity of a specific area of the brain stem, the rostral ventrolateral medulla (RVLM). It was further observed that mitochondrial dysfunction in cells from the RVLM was associated with sepsis-induced death.
Since coenzyme Q10 inhibits oxidative damage to lipids, proteins and DNA, it has potential to act as a neuroprotective agent in both acute and chronic brain damage. The efficacy of Ubisol-Aqua™: Coenzyme Q10 to act as a neuroprotective agent in sepsis was therefore assessed. In this study, sepsis was induced in animals leading to progressive hypotension and death. However, pre-treatment by microinjection of Ubisol-Aqua™: Coenzyme Q10 into the RVLM significantly prolonged survival time. This was associated with improvement in the blood pressure signal as well as blunting of the increased production of oxidants. These results support the potential efficacy of Ubisol-Aqua™: Coenzyme Q10 for disorders associated with an immunoinflammatory response, such as sepsis.
McCarthy S, Somayajulu M , Sikorska M, Borowy-Borowski H, Pandey S. Paraquat induces oxidative stress and neuronal cell death; neuroprotection by water-insoluble Coenzyme Q10. Toxicology and Applied Pharmocology 2004;201:21-31.
Paraquat is an extremely toxic herbicide, once widely used in North America. A strong association between paraquat exposure and Parkinson’s disease has been suggested. In this study performed in a cell model of human nerve cells, paraquat treatment induced mitochondrial dysfunction and ROS production leading to nerve cell death (apoptosis) similar to that seen in Parkinson’s disease. Pre-treatment of these cells with Ubisol-Aqua™: Coenzyme Q10 prior to paraquat exposure inhibited ROS generation and significantly reduced the number of dead cells. Treatment also increased energy (ATP) production and maintained normal cell biochemical function. These results provide evidence supporting the protective effects of coenzyme Q10 in nerve cell damage induced by environmental toxins.
Somayajulu M, McCarthy S, Hung M, Sikorska M, Borowy-Borowski H, and Pandey S. Role of mitochondria in neuronal cell death induced by oxidative stress; neuroprotection by coenzyme Q10. Neurobiology of Disease 2005;18:618-827.
Nerve cells depend heavily on mitochondrial production of ATP due to their high energy expenditure. They are also at risk for oxidative damage caused by the potentially toxic by-products of energy metabolism. Such oxidative stress can ultimately lead to cell death. It is believed that this mechanism plays a role in contributing to neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. In this study, two types of human nerve cells were grown in culture and exposed to hydrogen peroxide, which causes mitochondrial dysfunction. Mitochondrial dysfunction leads to an increase in reactive oxygen species (ROS) which cause oxidative damage to cell membranes, DNA, and ultimately cell death. When the cells were pretreated with Ubisol-Aqua™: Coenzyme Q10 and then exposed to hydrogen peroxide, total cellular ROS was decreased, the mitochondrial membrane remained intact and the nerve cells remained healthy. These studies suggest that water-soluble coenzyme Q10 has the potential to be used therapeutically for neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease.
Naderi J, Somayajulu-Nitu M, Mukerji A, Sharda P, Sikorska M, Borowy-Borowski H, Antonsson B, Pandey S. Water-insoluble formulation of Coenzyme Q10 inhibits Bax-induced destabilization of mitochondria in mammalian cells. Apoptosis 2006;8:1359-69.
This paper elucidates the mechanism by which coenzyme Q10 acts to prevent programmed cell death (apoptosis). Cultured human fibroblasts and embryonic kidney cells were pre-treated with Ubisol-Aqua™: Coenzyme Q10 and then subjected to hydrogen peroxide, a potent oxidant. The pre-treated cells showed a decrease in mitochondrial dysfunction and cell death compared to untreated cells. The mechanism by which this occurred involved the inhibition of a pro-apoptotic protein, Bax. These data further support the role of water-soluble coenzyme Q10 to prevent oxidative damage.
Mobarol M, Cherry CL, Wesselingh SL, Fain R and Hooker DJ. International AIDS Society Meeting, Sydney Australia 2007, Ubisol-Aqua™: Coenzyme Q10 prevents antiretroviral toxic neuropathy in an in vitro model.
Cultured fetal rat dorsal root ganglia (DRGs) have been established as an in vitro model of antiretroviral toxic neuropathy (ATN), a complication associated with AIDS treatment. ATN can occur in AIDS patients in countries where the use of nucleoside analogue reverse transcriptase inhibitors (NRTIs) stavudine (d4T) or didanosine (ddI) are used. Concentrations between 0.1 and 100 uM of Ubisol-Aqua™: Coenzyme Q10 (HQO™) were tested in this DRG model which measures neurite morphology and growth. All concentrations of HQO™ tested were associated with improved neurite growth (p<0.0001 versus control). DRGs cultured with HQO™ also remained viable for at least one week beyond those cultured under control conditions.
DRGs exposed to d4T demonstrated impaired neurite growth compared to controls (p = 0.01). When cells treated with d4T were exposed to HQO™, greater neurite growth was observed than those with d4T alone (p = 0.01) and were not different than controls. Similarly, when DRGs were exposed to ddI, neurite growth was significantly reduced compared to controls (p<0.001), and neurites were dead by day 16. This toxicity was completely prevented by HQO™ resulting in neurite morphology and growth that was the same as controls. These studies demonstrate that HQO™ enhances the growth and survival of fetal rat DRGs and reduces the toxicity of d4T and ddI in this in vitro model of ATN. These data support the possibility that coenzyme HQO™ may be useful for preventing ATN associated with ddI and d4T.
Lipshutz BH, Aguinaldo GT, Ghorai S, and Voigtritter K. “Olefin Cross-Metathesis Reactions at Room Temperature Using the Nonionic Amphiphile “PTS”: Just Add Water”, Organic Letters 2008, 10(7), 1325-1328.
This publication describes the first examples of unsymmetrical olefin cross-metathesis reactions in water as the only solvent. The reactions were conducted using water-insoluble substrates at room temperature, with commercially available catalysts, and small percentages of Zymes’ PTS. Nanometer micelles are formed in water that accommodate both substrates and Ru-based metathesis catalysts. The reactions proceed with high efficiency and very high E-selectivity, the products from which can be easily isolated.
Lipshutz .H and Taft BR “Heck Couplings at Room Temperature in Nanometer Aqueous Micelles”, Organic Letters 2008, 10(7), 1329-1332.
PTS, part of Zymes’ proprietary solubilization platform Ubisol-Aqua, has been shown to promote Heck cross-coupling between non-water-soluble reaction partners, in water, at room temperature. The formation of nanomicelles in water allows these first examples of Heck reactions in water (as the only solvent) at room temperature.
Lipshutz BH, Petersen TB, and Abela AR , “Room Temperature Suzuki-Miyaura Couplings in Water Facilitated by Nonionic Amphiphiles”, Organic Letters 2008, 10(7), 1333-1336.
Small percentages of PTS in water can be used to promote Suzuki-Miyaura cross- couplings. A dilute aqueous solution containing an arylboronic acid and one of a wide variety of aryl halides or pseudo-halides, in the presence of a Pd catalyst and inexpensive base, has been shown to afford biaryl cross-coupling products in good yields. In most cases, the reactions proceed at ambient temperatures, even with sterically hindered and lipophilic substrates.
Somayajulu-NiŃu M, Sandhu JK, Cohen J, Sikorska M, Sridhar TS, Matei A, Borowy-Borowski H, and Pandey S. Paraquat induces oxidative stress, neuronal loss in substantia nigra region and Parkinsonism in adult rats: Neuroprotection and amelioration of symptoms by water-soluble formulation of Coenzyme Q10. BMC Neuroscience 2009, 10:88 Epub.
Parkinson’s disease (PD), for which currently there is no cure, develops as a result of progressive loss of dopamine neurons in the brain; thus, identification of any potential therapeutic intervention for disease management is of a great importance. Prophylactic application of water-soluble formulation of coenzyme Q10 (PTS™: Coenzyme Q10) could effectively offset the effects of environmental neurotoxin paraquat, believed to be a contributing factor in the development of PD. This paper utilized a model of paraquat-induced dopaminergic neurodegeneration in adult rats. Histological and biochemical analyses of rat brains revealed increased levels of oxidative stress markers and a loss of approximately 65% of dopamine neurons in the substantia nigra region. The paraquat-exposed rats also displayed impaired balancing skills. In contrast, paraquat exposed rats receiving PTS™: Coenzyme Q10 in their drinking water prior to and during the paraquat treatment neither developed neurodegeneration nor reduced balancing skills and were indistinguishable from the control paraquat-untreated rats. This is the first preclinical evaluation of a water-soluble coenzyme Q10 formulation showing the evidence of prophylactic neuroprotection at clinically relevant doses.
Cherry CL, Mobarok M, Wesselingh SL, Fain R, Weinstock SB, Tachedjian G, Srivastava S, Tyssen DP, Glass JD,
Hooker DJ. Ubisol-AquaTM:Coenzyme Q10 Prevents Antiretroviral Toxic Neuropathy in an in vitro Model. In press, Current HIV Research.