Archive for the ‘Compound Screening’ Category
Families of Spinal Muscular Atrophy Funded Program Shows Quinazoline Compounds Give Survival Benefit in a Severe Mouse Model of SMA.
Last Updated on Monday, 28 December 2009 12:06 Written by Editor Monday, 28 December 2009 12:06
This publication, showing data from the testing of Quinazoline derivatives in a Spinal Muscular Atrophy mouse model, has been published in Human Molecular Genetics by lead author Dr. Matthew Butchbach from the laboratory of Dr. Arthur Burghes at the Ohio State University.
The generation of the Quinazoline compounds as a therapeutic drug candidate for Spinal Muscular Atrophy was fully funded by Families of SMA.
The paper explores whether the Quinazoline compounds, which increase the expression of SMN2, are useful as potential therapeutics for SMA. Ultrahigh-throughput screening identified substituted Quinazolines as potent SMN2 inducers. The drug-like properties of the initial screening hits were optimized through directed medicinal chemistry. This resulted in series of C5-Quinazoline derivatives.
Oral administration of three of these compounds (D152344, D153249 and D156844) to neonatal mice resulted in a dose-dependent increase in Smn promoter activity in the central nervous system.  The authors then examined the effect of these compounds on the progression of disease in SMNDelta7 SMA mice. Oral administration of D156844 significantly increased the mean lifespan of SMNDelta7 SMA mice by approximately 21-30% when given prior to motor neuron loss. Overall the authors summarize that the quinazoline derivative D156844 increases SMN expression in neonatal mouse neural tissues, delays motor neuron loss at PND11, and ameliorates the motor phenotype of SMNDelta7 SMA mice.
“This is the first compound series to go from hit-to-preclinical candidate that shows favorable pharmacology in the nervous system and shows benefit to severe SMA mice. This study shows that promising therapies for SMA can be developedâ€, said Matthew Butchbach, Ph.D., who is lead author on this publication.
“Families of SMA is pleased that the first test of this class of compounds in SMA mice shows potential therapeutic benefit. The clinical lead in this series called Quinazoline495, which is a more optimized compound than those tested here, has also been assessed in this animal model with similar results, as well as tested in a slightly less severe mouse model of SMA, in which it showed marked enhancement of survival”, says Jill Jarecki, Ph.D., FSMA research director.
The lead compound Quinazoline495 recently received orphan drug designation from the FDA for the treatment of spinal muscular atrophy. Please click here to read more.
Families of SMA recently licensed this series of compounds to Repligen Corporation for development as a drug treatment for Spinal Muscular Atrophy.
The full reference:
Butchbach ME, Singh J, Thornorsteinsdóttir M, Saieva L, Slominski E, Thurmond J, Andrésson T, Zhang J, Edwards JD, Simard LR, Pellizzoni L, Jarecki J, Burghes AH, Gurney ME. Effects of 2,4-diaminoquinazoline derivatives on SMN expression and phenotype in a mouse model for spinal muscular atrophy. (2009). Human Molecular Genetics, Epub ahead of print.
Source: fsma.org
Posted under Compound Screening, Drug-Like Compounds, Press Releases | Comments Off
Enzyme binds both sides of the mirror
Last Updated on Monday, 28 December 2009 11:30 Written by Editor Monday, 28 December 2009 11:30
European chemists have discovered that both mirror-image forms of a particular compound can bind at the same time in the same site of an enzyme, a phenomenon that has never been seen before. The finding has significance for drug discovery screening and studies of how small molecules interact with proteins.
Rolf Breinbauer from Graz University of Technology, Austria, and Wulf Blankenfeldt from the Max Planck Institute of Molecular Physiology in Dortmund, Germany, were studying a metabolic enzyme from a species of the bacterium Burkholderia cepacia, using racemic mixtures of chiral probe molecules to find ones that bound in the enzyme’s active site. In most cases only one form of a chiral (or ‘handed’) molecule would bind at once, but they found that in one instance both enantiomeric forms occupied the binding site at the same time.
‘If you read the textbooks about enantiomers,’ says Breinbauer, ‘there’s a simplified notion that one enantiomer is good and the other is either bad or just idle.’ He explains that for most proteins (apart from certain enzymes that have evolved to cope with wide ranges of substrate molecules) either only one enantiomer will bind, or both can bind individually - with the assumption that one form will be significantly more active than the other. ‘Our findings show that the world is more complicated,’ he adds.
While each individual enantiomer can bind to the enzyme seperately, Breinbauer notes that the arrangement of the molecules within the binding site is quite different when both bind together. This could lead to cooperative effects, producing either an enhanced or diminished response relative to the individual enantiomers.
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The three ways enantiomers can bind in enzymes: only one enantiomer binds (top); each binds individually (middle); both bind together (bottom)
© Angewandte Chemie
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He adds that this could have relevance in drug discovery screening, where mixtures of both enantiomers of chiral compounds are routinely screened together to find initial hits. ‘People need to consider more options when interpreting binding data from racemic mixtures.’
Dafydd Owen of Pfizer Research Chemistry in Sandwich, UK, agrees that the finding is an important reminder that chemists need to be open-minded about interpreting screening data. It also highlights the inherent trade-offs made when screening mixtures - particularly in high-throughput screens when mixtures of several compounds are tested at once.
Owen sees most interest in the discovery in the area of fragment-based drug discovery, where small ‘fragment’ molecules found to bind to a drug target are linked together to make potential drug molecules. ‘As a medicinal chemist,’ he adds, ‘my immediate thought was to join the two structures together to incorporate the best of each and make a hybrid.’ He points out, however, that from a fragment point of view it is almost irrelevant to the enzyme that the two molecules happen to be mirror images of each other, ‘despite their apparent similarity, nature views enantiomers as very different molecules’.
Phillip Broadwith
Source: rsc.org
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UT professor receives grant for new process
Last Updated on Wednesday, 16 December 2009 10:59 Written by Editor Wednesday, 16 December 2009 10:59
Using a pair of tweezers, a UT graduate student carefully lifted a nylon mesh square about the size of a thumbnail out of a small flask in his team’s lab.
The nylon had been soaking in a clear, watery solution containing a chemical compound — the “capture agent†— that it would bind with during a process tweaked by the student and his team.
Jennifer Brodbelt, a chemistry and biochemistry professor, received a $734,068 grant from the National Institutes of Health Oct. 1 to further develop the process — Desorption Electrospray Ionization (TM-DESI) — and perfect the nylon squares which isolate desired compounds from solutions.
Brodbelt, UT graduate students and two professors from Southwestern University in Georgetown were given a two-year deadline to gather blood from people of varying ages and levels of health, and to develop a more efficient method of analyzing the samples.
Results will be used to spot trends in the frequency of certain biological compounds, including amino acids.
A mass spectrometer, the machine Brodbelt’s team uses, can identify specific compounds in a mixture like blood. The tricky part was getting the sample to spray into the machine.
Joe Chipuk, a graduate student currently working on the project, was struck by the idea of having samples sprayed directly through a sifting material into the spectrometer.
Chipuk ran home and began collecting mesh materials to spray water through. He cut up his screen door, his wife’s tea strainer and the aerator from his kitchen faucet.
He went outside, used a hose to spray water through the mesh materials and observed the water’s exit path.
He then drafted a plan to create a mesh material soaked in a chemical that allows certain compounds to travel through but traps enough as to not let every metabolite escape.
After the unwanted materials are sorted out, the desired compounds attached to the mesh are released and analyzed by the spectrometer.
Before Chipuk’s square, the desorption process played out very much like a complicated billiards shot. The spray came down at an angle, hit the slide holding the blood sample and ricocheted off carrying the compounds through the spectrometer.
The new technique allows the compounds to be sorted and analyzed at a much faster pace than before. Chipuk said they can now analyze 50 samples in approximately eight minutes, whereas before, analyzing 50 samples would have taken more than 24 hours.
The team is focusing on improving the reliability and consistency of the mesh squares, Brodbelt said.
“The hope is that this could be a way to diagnose patterns of disease or determine a prognosis based on the pattern of metabolites,†Brodbelt said. “The sooner you have an idea that you might have cancer, or that you are on track to develop cancer, you could have screening done earlier and more frequently.â€
Source: dailytexanonline.com
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Computational Method Points To New Uses, Unexpected Side Effects Of Already Existing Drugs
Last Updated on Tuesday, 15 December 2009 04:19 Written by Editor Tuesday, 15 December 2009 04:19
Scientists at the University of North Carolina at Chapel Hill School of Medicine and the University of California, San Francisco have developed and experimentally tested a technique to predict new target diseases for existing drugs.
The researchers developed a computational method that compares how similar the structures of all known drugs are to the naturally occurring binding partners — known as ligands — of disease targets within the cell. In a study published this week in Nature, the scientists showed that the method predicts potential new uses as well as unexpected side effects of approved drugs.”This approach uncovered interactions between drugs and targets that we never could have predicted simply by looking at the chemical structures,” said senior study author Bryan Roth, M.D., Ph.D., professor of pharmacology and director of the National Institute of Mental Health Psychoactive Drug Screening Program at UNC. “We may now have a way to predict what side effects are likely to occur from treatment before we even put a drug into clinical testing.” Roth is also a member of the UNC Lineberger Comprehensive Cancer Center.
Many of the most successful drugs on the market today are being prescribed for ailments that are quite different from the ones they were originally designed to treat. Viagra, for instance, was once intended for coronary heart disease but now is used to combat erectile dysfunction. The discovery of surprising uses of developed drugs can sometimes be the result of serendipity, as unforeseen side effects emerge from clinical trials. In the past, researchers have tried to predict drug interactions by looking for chemical similarities among the possible targets of pharmaceutical compounds.
However, some drug targets which look very similar to one another bind very different ligands, and some targets that don’t have any obvious similarity bind similar ligands, says Brian Shoichet, Ph.D., co-senior study author and professor of pharmaceutical chemistry at the University of California at San Francisco. “So if instead we were to organize targets by the ligands they recognize, it could reveal different patterns than traditional approaches, and illuminate new opportunities for drugs to bind to unexpected targets.”
A team of researchers led by Roth and Shoichet did just that, comparing the structures of 3,365 FDA-approved and investigational drugs against the structures of hundreds of targets, defining each target by its ligands. They then honed in on thirty of the strongest predictions, validating the actual physical interactions between the drugs and targets in wet laboratory experiments.
In one of their follow-up experiments, the scientists investigated the molecular targets of the hallucinogenic substance dimethyltrytamine (DMT), which had previously been postulated to act through a site known as the sigma-1 receptor. Using the computational approach, Roth and colleagues found that DMT had a high affinity for serotonin receptors, including the binding site for LSD, another hallucinogen.
They also showed that the substance is hallucinogenic in normal mouse models but not in ones lacking the serotonin receptor. Roth says the power of their approach is it can be used to uncover the real targets of pharmaceutical compounds quickly and efficiently, and will probably lead to a greater understanding of the many molecular targets of each drug.
“Drugs are not as selective as we once thought,” said Roth, who is also a professor in the School of Pharmacy’s medicinal chemistry and natural products division. “It turns out that the most non-selective drugs are frequently the most effective for complex diseases. Rather than ‘magic bullets,’ we need to come up with ‘magic shotguns’ that hit more than one molecular target at a time. We could use this computational approach to identify the drugs that hit the right targets and miss the wrong ones.”
Study co-authors from UNC include Vincent Setola, research associate professor; Atheir Abbas, former graduate student; Sandra J. Hufeisen, senior research assistant; Niels H. Jensen, research associate; Michael B. Kuijer, research technician; Roberto C. Matos, research technician; Thuy B. Tran, research technician; Ryan Whaley, research technician; and Richard A. Glennon. The paper’s first author is Dr. Michael Keiser, from the UCSF side of the collaboration. Also from UCSF were Drs. John Irwin, Christian Laggner and Jerome Hert, and PharmDs Kelan Thomas and Douglas Edwards.
Funding for the studies at UNC and at UCSF came from the National Institutes of Health.
ScienceDaily (Nov. 7, 2009)
Posted under Compound Libraries, Compound Screening, Drug Development, Press Releases | Comments Off
Professor Receives Grant to Develop More Rapid Technology For Screening Blood Samples
Last Updated on Tuesday, 15 December 2009 03:29 Written by Editor Tuesday, 15 December 2009 03:29
AUSTIN, Texas — Dr. Jennifer Brodbelt, professor of chemistry and biochemistry at The University of Texas at Austin, has received a $734,068 grant from the National Institutes of Health (NIH) to develop a new method for rapidly screening blood samples for biomarkers.
Biomarkers are small molecules that indicate the presence of a particular physiological condition, typically a disease. The new method, if successful, could prove useful not just for identifying markers of specific diseases such as cancer or heart disease, but for discovering broader metabolic patterns correlated with conditions such as aging or obesity.
“There are technologies right now that are very effective at separating and analyzing the different compounds in a blood sample, but they tend to be relatively slow,” says Brodbelt, the principal investigator of the grant. “It makes it very hard to do analyses of lots of samples. What we’re developing is a chip-based method, where entire classes of compounds are captured on the chips and then all the compounds are released and analyzed by mass spectrometry in just a few seconds.”
Although the technology, if successful, should be useful in searching for biomarkers in all sorts of conditions, Brodbelt and her collaborators from Southwestern University in Georgetown, Texas, Drs. Lynn and Frank Guziec, are focusing on patterns that correlate with aging.
“We’re trying to develop maps that can correlate the progression of aging with metabolites that might be circulating in your blood,” says Brodbelt. “These could be small molecules that increase in quantity as you age, or actually change in composition as one ages.”
The new method, says Brodbelt, involves three basic stages.
The first stage is the coating of different regions of a mesh chip with a variety of “capture agents,” which chemically bind to specific compounds in a blood sample. A burst from an ultraviolet light then severs the chemical bonds between the chip and the captured substances. Then an electrospray, which is similar to a solvent aerosol spray, shoots through the mesh chip and transfers the different compounds into the mass spectrometer for analysis.
By analyzing the mass spectrometric data, says Brodbelt, scientists should be able to measure the presence and quantity of different compounds, and to do so on a scale, and with a speed, that wasn’t possible before.
“The payoff could be big,” she says. “It’s a different strategy than what might be pursued by molecular biologists or biochemists. They’ll often focus on studying one or two proteins at a time, and develop a really deep understanding of those proteins. We’re looking for the more generalized profile, and we may notice some patterns that weren’t apparent to them.
“There are so many other areas where you’d want to do profiling. It might involve looking for pesticides as part of an environmental study, or doing protein-related work or drug profiling work. If this approach is successful, I imagine other groups will try to develop these chips as well.”
Brodbelt’s grant, which is being funded as part of the American Recovery and Reinvestment Act (ARRA) government stimulus package, is a “Challenge Grant,” meant to encourage high-risk, high-reward research projects that may produce results quickly.
This is the second NIH grant in two years that Brodbelt and the Guziecs have received. In 2008, the collaborating groups received a four-year, $1,113,615 grant to evaluate an innovative technique that could assess the anti-cancer activity of new compounds.
For more information, contact: Jennifer Brodbelt, Department of Chemistry and Biochemistry, 512-471-0028.
Posted under Compound Screening, Grants and Awards, Press Releases | Comments Off
Antiviral activity of gliotoxin, gentian violet and brilliant green against Nipah and Hendra virus in vitro
Last Updated on Tuesday, 15 December 2009 03:27 Written by Editor Tuesday, 15 December 2009 03:27
Using a recently described monolayer assay amenable to high throughput screening format for the identification of potential Nipah virus and Hendra virus antivirals, we have partially screened a low molecular weight compound library (>8,000 compounds) directly against live virus infection and identified twenty eight promising lead molecules. Initial single blind screens were conducted with 10 microM compound in triplicate with a minimum efficacy of 90% required for lead selection.
Lead compounds were then further characterised to determine the median efficacy (IC50), cytotoxicity (CC50) and the in vitro therapeutic index in live virus and pseudotype assay formats.
Results: While a number of leads were identified, the current work describes three commercially available compounds: brilliant green, gentian violet and gliotoxin, identified as having potent antiviral activity against Nipah and Hendra virus. Similar efficacy was observed against pseudotyped Nipah and Hendra virus, vesicular stomatitis virus and human parainfluenza virus type 3 while only gliotoxin inhibited an influenza A virus suggesting a non-specific, broad spectrum activity for this compound.
Conclusions: All three of these compounds have been used previously for various aspects of anti-bacterial and anti-fungal therapy and the current results suggest that while unsuitable for internal administration, they may be amenable to topical antiviral applications, or as disinfectants and provide excellent positive controls for future studies.
Author: Mohamad AljofanMichael SgangaMichael LoChristine RootesMatteo PorottoAdam MeyerSimon SaubernAnne MosconaBruce Mungall
Credits/Source: Virology Journal 2009, 6:187
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Scientists hope mouse research leads to new anti-cancer therapies
Last Updated on Wednesday, 2 December 2009 12:28 Written by Editor Wednesday, 2 December 2009 12:28
Recent collaborative work between Cambridge, Mass., research institutes has discovered a method of screening for chemicals that selectively kill breast cancer stem cells in culture and in mice, a breakthrough that may directly or indirectly lead to new anti-cancer therapies.
“One of the major difficulties with developing good anti-cancer drugs is that the anti-cancer drugs don’t cure the tumors, and part of the reason they don’t cure the tumors is that they’re not very effective in specifically attacking and eliminating the cancer stem cells,†said Dr. Robert Weinberg, founding member of the Cambridge-based Whitehead Institute and a biology professor at MIT. “We’re trying to develop techniques to understand what creates cancer stem cells and how they are perpetuated.â€
A theory prevalent amongst many researchers suggests that the aggressive subset of cancer cells — called cancer stem cells — drives tumor growth and causes tumors to regenerate after chemotherapy has killed 99 percent of their cells.
Isolating true-to-form cancer stem cells proved to be a challenge until recently, when researchers at Weinberg’s lab at the Whitehead Institute discovered a method to manipulate these cells.
The discovery allowed a team of scientists led by Dr. Piyush Gupta of the Cambridge-based Broad Institute to derive cell lines from human breast epithelial cells and use them to screen 16,000 chemicals in culture dishes. The scientists found that 32 of these chemicals specifically target cancer stem cells and kill them.
Gupta’s team then tested the chemical compounds in mice, and narrowed the results down to one chemical, salinomycin, that appeared to shrink tumor growth. The study was published in an August issue of the journal Cell.
“Ours was really the first step in a long process.†Gupta said. “We have one compound now, and it is not clear whether this one compound is ideal in terms of its activity and also in terms of its toxicity.â€
The discovery of the potent chemical does not necessarily mean that it will have any improvement over current cancer treatment. Gupta’s team has started the extensive follow-up testing that is necessary to determine in which stage eliminating cancer stem cells would be most beneficial to patients, and whether the compound is suitable for humans at all.
“Things sometimes appear very promising in pre-clinical studies but then in patients they may for whatever reason not work as well,†Gupta said. “All we can do is try to design the best possible preclinical studies in the hope that it will work in the patients. We really want to understand how the compounds work in animals before we even think about putting them in people.â€
Still, the new screening method is a promising development in the field of anti-cancer treatment. Gupta said he expects some cancer stem cell-targeting therapies to make into human trials within three to five years.
“For a while it seemed like these cancer stem cells were exciting, but there was very little known about them,†Gupta said. “Now I think that we’re finally at the stage where we can really start to understand what’s going on inside these cells.â€
Wicked Local Cambridge
Source: tauntongazette.com
Posted under Cancer Research, Compound Screening, Discoveries, Innovations and Patents, Oncology Research, Press Releases | Comments Off
Galapagos expands strategic alliance in metabolic disease with Merck & Co., Inc
Last Updated on Tuesday, 1 December 2009 12:55 Written by Editor Tuesday, 1 December 2009 12:55
* Galapagos to discover small molecule candidate drugs for pre-clinical development in atherosclerosis* Total alliance milestones more than double to over EUR 400 million, plus royalties on worldwide sales
Mechelen, Belgium 14 October 2009 – Galapagos NV (Euronext: GLPG) announced today that it has expanded its global strategic alliance in metabolic diseases with an affiliate of Merck & Co., Inc to incorporate the development of new therapies for atherosclerosis.
Galapagos will be responsible for the discovery and pre-clinical development of new small molecule candidate drugs based on novel Galapagos targets. The alliance will make use of Galapagos’ proprietary target discovery platform for identification of novel targets in atherosclerosis, as well as in obesity and diabetes. After validation, targets will be selected by a joint steering committee and entered into screening and chemistry by Galapagos. Merck will have an exclusive option to license in each candidate for clinical development and commercialization on a worldwide basis. Upon exercise of such option, Merck will be responsible for the development and commercialization of the candidate drug. Galapagos may execute Phase I clinical studies and will have the right to further develop and commercialize certain compounds for which Merck does not exercise its exclusive option.
In January 2009, Galapagos announced an alliance with Merck in diabetes and obesity, with milestone payments with the potential to exceed EUR 170 million. Under the terms of this expanded agreement, that now includes small molecule candidate drugs for pre-clinical development in atherosclerosis, Galapagos is eligible to receive research, regulatory and sales milestone payments that may total in excess of EUR 400 million. In addition Galapagos is eligible to receive royalties upon commercialization of any products covered under the agreement.
The expansion announced today is separate from Galapagos’ alliance with Merck in inflammatory diseases announced in April 2009.
“We are pleased to expand our relationship with Merck, a highly synergetic and decisive partner in the metabolic and inflammation alliances,” said Onno van de Stolpe, CEO. “Today’s expansion into atherosclerosis fits very well into our strategy of leveraging the novelty coming out of Galapagos’ R&D approach.”
“Merck’s extended collaboration with Galapagos underscores our ongoing commitment to identifying novel therapeutic targets for fighting cardiovascular and metabolic diseases,” said Catherine Strader, Vice President, External Basic Research, Merck Research Laboratories.
About atherosclerosis
Atherosclerosis, commonly referred to as hardening of the arteries, involves the thickening of the artery walls as a result of the build-up of fatty materials such as cholesterol. Atherosclerosis is considered the most common underlying cause of strokes, heart attacks, and most cardiovascular diseases in general, including congestive heart failure. Treatment includes lifestyle changes and medicines. Global sales of cardiovascular drugs exceeded $95 billion in 2008, making it the largest therapeutic area worldwide. Current research is focused on reducing side effects and costs for new atherosclerosis therapies.
About Galapagos
Galapagos (Euronext: GLPG; OTC: GLPYY) is a drug discovery and development company with small molecule programs in bone and joint diseases, bone metastasis, cachexia, anti-infectives, atherosclerosis, and metabolic diseases. It has established risk sharing alliances with GSK, Janssen Pharmaceutica, Eli Lilly and Merck & Co., Inc. Through an alliance with MorphoSys, Galapagos is also developing new antibody therapies in bone and joint diseases. Its division BioFocus offers a full suite of target-to-drug discovery products and services to pharmaceutical and biotech companies and to patient foundations, encompassing target discovery and validation, screening and drug discovery through to delivery of pre-clinical candidates. Galapagos currently employs 495 people and operates facilities in six countries, with global headquarters in Mechelen, Belgium. More info at: www.glpg.com
CONTACT Galapagos NV Onno van de Stolpe, CEO Tel: +31 6 2909 8028
ir@glpg.com
This release may contain forward-looking statements, including, without limitation, statements containing the words “believes,” “anticipates,” “expects,” “intends,” “plans,” “seeks,” “estimates,” “may,” “will,” “could,” “stands to,” and “continues,” as well as similar expressions. Such forward-looking statements may involve known and unknown risks, uncertainties and other factors which might cause the actual results, financial condition, performance or achievements of Galapagos, or industry results, to be materially different from any historic or future results, financial conditions, performance or achievements expressed or implied by such forward-looking statements. Given these uncertainties, the reader is advised not to place any undue reliance on such forward-looking statements. These forward-looking statements speak only as of the date of publication of this document. Galapagos expressly disclaims any obligation to update any such forward-looking statements in this document to reflect any change in its expectations with regard thereto or any change in events, conditions or circumstances on which any such statement is based, unless required by law or regulation.
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Source: pr-usa.net
Posted under Collaborations, Compound Screening, Drug Development, Press Releases | Comments Off
High-Content Screening Surges Ahead
Last Updated on Tuesday, 1 December 2009 11:33 Written by Editor Tuesday, 1 December 2009 11:33
High-content screening (HCS) and the technology to do it faster, on more compounds in a shorter period of time, and to generate quantitative, multiparametric data took center stage at CHI’s “High Content East†meeting held in Boston last month. Presenters described how they are implementing enhanced screening systems, image-analysis methods, and data-management strategies to achieve daily HCS runs on tens of thousands of wells and screening campaigns totaling 200,000 to 3 million wells.
High throughput HCS—albeit not yet reaching the numbers common for conventional high-throughput screening (HTS) and with lingering limitations and challenges related to live-cell imaging over time—is making its mark and being used to probe the biological basis of disease and to detect even subtle phenotypic changes in response to experimental compounds.
Determining whether a cell looks like a cancer cell, for example, typically requires being able to detect subtle morphological changes, such as small alterations in size or structure, changes in the connections a cell makes with neighboring cells, or variations in the texture of staining. These have, historically, been mainly qualitative parameters detected by studying and comparing images of cells.
In her talk at the conference, Anne Carpenter, Ph.D., director of the imaging platform at the Broad Institute of Harvard University and MIT, presented her group’s work using HCS and image analysis to quantify difficult phenotypes and differentiate disease states such as leukemia.
Not only do HCS systems and image-analysis software automate the screening process, enabling theanalysis of many more cells in less time and increasing the chances of detecting even small numbers of altered cells, they can also utilize algorithms that evaluate defined combinations of parameters in a quantifiable manner and apply techniques to distinguish between clumping or closely juxtaposed cells. Relying on computer-based image analysis also standardizes the process, eliminating factors such as variability in human expertise and experience, consistency, and fatigue.
Dr. Carpenter’s group uses machine-learning methods to train image-analysis software to identify subtle phenotypic changes. Biologists work with the software in an iterative fashion in a process called supervised machine learning. They teach and correct the computers on a series of test images, refining the system’s knowledge base in a process that typically takes less than a day. The group developed the algorithms used by the biologists and has made them available as open-source software.
A recent paper published in PNAS by T. R. Jones, et al., documents the use of a trained image-analysis system to discriminate 15 different cellular phenotypes. Other projects involve teaching the software to discriminate leukemic from normal cells, to identify liver cells that are growing normally in culture—to aid in the development of physiologic models of liver function for use in drug testing—and training computers to detect subtle changes that signal the initiation of cell division for studying cell-cycle regulation in cancer.
Neil Carragher, Ph.D., senior scientist in the advanced science and technology laboratory at AstraZeneca, described how the company is applying high-content and live-cell imaging techniques and integrating the results with data derived from in vivo imaging and proteomic studies to improve clinical predictability.
Dr. Carragher’s group combines the results of high-content in vitro and in vivo assays to generate mechanistic information about phenotypic responses on candidate therapeutic compounds. The goal is to create a multiparametric fingerprint of a phenotype from images generated by HCS and to use this knowledge to enhance predictions of efficacy and toxicity early in drug discovery and reduce attrition later in development.
The phenotypic signatures are based on measurements of approximately 150 different parameters per cell for each assay. Data from multiple assays is collated for every test compound and compared with data obtained using well-characterized reference compounds to generate mechanistic hypotheses.
Only recently has open-source and commercial software become available “that allows you to quantitate more complex phenotypes, subtle changes, and heterogeneous responses from images,†Dr. Carragher said.
His group is employing two main approaches—each with different advantages and limitations. The first strategy relies on Definiens’ Cognition Network Technology™ software that allows users to develop algorithms that capture, computationally, what researchers can see visually. “It is very much context-based†and identifies objects based on how they are related to others in the image, rather than as individual pixels, explained Dr. Carragher. The in-house algorithm-development process depends on iterative programming steps. The other approach involves machine-learning tools using software such as the CellProfiler developed at the Broad Institute.
Redirecting Approved Drugs
| Identifying new applications for FDA-approved drugs using HCS and image-based systems biology is the focus of work being done by Stephen Wong, Ph.D., founding director of the bioinformatics and biomedical engineering program and the cellular and tissue microscopy core at the Methodist Hospital Research Institute and professor of radiology and neurosciences at Weill Cornell Medical College.
Dr. Wong gave examples of screening campaigns to decipher targets in the pathways responsible for the metastasis of breast cancer to the brain in his talk. He specifically described the computational tools his group is developing for high-content and network analysis, and the animal-imaging techniques being used to evaluate combinations of small molecule chemotherapeutic agents for their ability to cross the blood-brain barrier and to have an effect against central nervous system metastases in breast cancer. Dr. Wong’s group has also developed a series of quantitative image-analysis tools, including zebrafish image quantifier (ZFIQ), as well as software for studying neuronal spines (NeuronIQ), neurites (Neurite IQ), and time-lapse mitotic events in cells (DCellIQ). Dr. Wong’s HCS/systems biology research is funded by the NCI, NIA, and NLM. Because the compounds being studied are already approved drugs, Phase I trials are not needed. The quantitative data generated from HCS provides the evidence necessary for moving into Phase II studies, shortening the drug-development cycle to a year or less. The types of studies essential to Dr. Wong’s efforts, such as assays to monitor cell-cycle regulation or dendritic spine dynamics, require time-lapse, live-cell imaging. Looking at fixed cells provides only an artificial snapshot of where cells are at a particular point in time, explained Dr. Wong. “We want to look at a 384-well plate of continuously growing cells over five to six days,†he said, and in his view none of the instrument manufacturers competing in the HCS market has yet to provide a robust, incubator-based, environmentally controlled system that can achieve this. Vendors have tended to view HCS as just another type of high-throughput screening, but live-cell imaging done in as natural an environment as possible has quite different requirements, contended Dr. Wong. “Vendors are going in the wrong direction. The power of HCS is in the ability to visualize things in action and to extract lots more quantitative information from the images. If you, instead, retrofit HCS to HTS, you are losing its advantages,†such as the ability to see cells or spines change over time, to visualize cell-cell interactions, and to sync cell populations and study cell-cycle events in time-lapse, said Dr. Wong. In any experiment, “if you generate enough data you will get hits, but how many will be real hits versus false positives?†asked Dr. Wong. “We need to push the quality upfront on the biology side†and screen out, earlier in the discovery process, compounds that are destined to fail. Researchers at Pfizer are using HCS to study the genetic variation and physiologic interactions that underlie hepatic insulin resistance in type 2 diabetes and the prediabetic state. Diabetes is a complex, multigenic disease, and while advances in genomic and SNP-based technologies have led to the identification of at least 30 genes that contribute to the diabetic phenotype, much work remains to understand their role in cell biology and disease and how they interact. “If you are careful about the cell models you choose, you can use HCS to characterize these genes and monitor their effects on biochemical pathways,†said Steven Haney, Ph.D., associate fellow in biological profiling at Pfizer’s biotherapeutics and bioinnovation center. The company has invested heavily in developing cell models that are representative of human physiology, including hepatocytes that faithfully mimic liver function when grown in culture. The other main aspect of this research effort involves identifying changes that affect the diabetic phenotype, specifically glucose storage and utilization pathways, and distinguishing between effects that involve the insulin-signaling pathway from more general phenomena related to activation of toxicologic or stress pathways. “HCS can alert us to things we don’t necessarily know to look for, in a mechanism-independent way,†said Dr. Haney. “The increasing throughput of HCS allows us to look at a lot of cells and determine whether subtle phenotypic changes are significant or spurious.†Vendors Roll Out Image-Analysis SolutionsVersatility across application areas, from microscope-based imaging for detecting intracellular phenomena to high-speed scans at the cellular level to whole organism screening, is the focal point of instrument development at MDS Analytical Technologies. “With the options in our Complete Solution and the right infrastructure, you can use image-based assays for primary screening. We have tackled all the common bottlenecks,†said Michael Sjaastad, Ph.D., director of marketing for cellular imaging at MDS. The IsoCyte® DL laser-scanning cytometer complements the company’s ImageXpress® instrument platform as part of its overall HCS solution. MDS offers a high-throughput option that can screen and do image analysis on a 1,536-well plate in two to five minutes, according to Dr. Sjaastad. The instrument can image whole wells for accurate cell counting in cell-viability measurements, scan a microscope slide, or produce and analyze images of organisms such as zebrafish or worms when used in conjunction with the MetaXpress image-analysis software. For now, current systems “have the image resolution and acquisition speed researchers need,†and in Dr. Sjaastad’s view, future improvements will focus on “streamlining the data-analysis workflow and bringing the costs down per data point.†In a workshop at the meeting, Oliver Leven, Ph.D., head of screener professional services at Genedata, identified several ongoing challenges in HCS, including managing the volume and complexity of the data, improving the efficiency of data analysis, and creating an audit trail of results interpretation. As the throughput and scale of HCS increases, so too, do the difficulty and scope of these challenges. As researchers scale up an assay for high-throughput HCS, they need to select a defined set of parameters that represent the phenotype of interest and that allow them to assess the quality of both the assay and the data output. They also need to identify threshold values above or below which a result signifies a change in phenotype. The typical HCS image-analysis software that drives HCS systems routinely quantifies the cell images to yield a numerical description of the phenotypes. For large experiments, however, Dr. Leven described the researcher’s need to go back and view an image associated with an interesting or suspicious measurement as a persistent bottleneck. “The image is the experiment,†said Dr. Leven. A hit should signify a change in the cells, but it could also be an anecdotal finding or the result of an image out of focus. Distinguishing true hits from false positive results remains a challenge. Dr. Leven recounted the HCS projects that Genedata has performed for its pharma customers emphasizing the ability of the company’s High Content Analyzer—a new addition to the Genedata Screener® enterprise solution—to retrieve immediately any desired image. The high-throughput HCS projects described by Dr. Leven were able to analyze 40,000 compounds on a daily basis, for a total campaign of more than two million compounds, generating multifeatured data sets for each well. PerkinElmer’s high-content screening portfolio includes the Opera confocal microplate image reader and Acapellaâ„¢ image-analysis software, the compact Operetta HCS system, driven by Harmonyâ„¢ software, and the Columbusâ„¢ data-management system and new Columbus 2.0 for use with the Opera platform. Gabriele Gradl, Ph.D., global product leader for HCS at PerkinElmer Cellular Technologies, emphasized the complexity involved in deriving robust, quantitative data from cellular measurements derived on image analysis of high-content screens. Whereas, fluorescence-based analysis typically relies on identifying objects in cells and measuring their fluorescence intensities, PerkinElmer has developed a computational strategy that is independent of absolute fluorescence intensity. It relies on texture analysis and quantitative pattern analysis for data generation. Texture-analysis tools can detect patterns and effects that would not be apparent on routine visual analysis, according to Dr. Gradl. Threshold adjacency statistics is one example of such a tool. It searches for differences in fluorescence intensity values between adjacent pixels over a defined distance. Dr. Gradl described the particular advantages of applying texture analysis for detecting subtle morphologic changes associated with cell viability or toxicity assays and in stem cell research. It can detect differences not visible to the eye and identify changes that the user might not even have known to look for in the data. She presented, as an example, the use of texture analysis to assess mitochondrial integrity, as loss of mitochondrial activity and enhanced mitochondrial biogenesis are early markers of cytotoxicity. Dr. Gradl also described the use of texture analysis in brightfield imaging and the ability to assess segmentation based on granularity, enabling label-free proliferation assays and analysis of cell differentiation in real time. The algorithms developed by PerkinElmer can apply texture analysis to whole cells or to specific intracellular compartments depending on the design of the assay. The company is exploring a range of applications for its texture-analysis software tools, including stem cell differentiation analysis, quality control of stem cells produced for therapeutic use, live-cell imaging over time, and 3-D tissue sample analysis. Earlier this year, GE Healthcare introduced the IN Cell Analyzer 2000 cell-imaging system, which incorporates several new features: preview scoring of a selected area of a sample before an acquisition run; a large chip CCD camera coupled with a widefield illumination source for twice the brightness of a conventional xenon lamp, according to GE; whole-well imaging; an objectives range from 2x–100x; six imaging restoration modes; and a manual microscope mode. Fred Koller, Ph.D., president and CEO of Cyntellect, launched the company’s new Celigoâ„¢ cytometer at the “High Content East†meeting, emphasizing the system’s ability to image “every cell in every well,â€Â from edge to edge without edge effects using both brightfield and fluorescence imaging. Cyntellect’s optical technology achieves high-quality large field imaging using a set of mirrors to capture each well in its entirety without moving the plate and without the need to refocus, allowing for rapid, full-plate imaging. Celigo provides “uniform illumination with no gradient across the well,†said Dr. Koller, and allows for a combination of label-free imaging and three-color fluorescence. He described the instrument’s capabilities for performing cell-counting assays, cell growth tracking, and confluency studies, for example, and for noninvasive imaging of stem cell cultures without disrupting their three-dimensional colony structures. Celigo can switch from single-cell to colony-counting mode. The company has also developed a secretion assay for use on the Celigo that measures the amount of protein secreted by individual cells. The assay can distinguish between high and low secretors and is useful for detecting heterogeneity and instability in cell cultures early in process development, such as for antibody manufacturing. The Cellular Imaging and Analysis group at Thermo Fisher Scientific introduced the Cellomics iDevâ„¢ intelligent assay development workflow for HCS image analysis at “High Content Eastâ€. Users work training image sets of positive and negative biology, applying imaging and analytical algorithms that allow for real-time interaction with the images. The software employs the biological data generated to optimize assay protocols. Source: genengnews.com |
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Evotec, Ono Extend Drug Discovery Services Pact
Last Updated on Tuesday, 1 December 2009 11:29 Written by Editor Tuesday, 1 December 2009 11:29
The companies began their drug target collaboration in March 2008.
Under the agreements, Evotec will provide high-throughput screening, in vitro pharmacology, protein crystallography, and medicinal chemistry services to discover small molecular weight compounds that will be used against an ion channel target. The aim of the collaboration is to move Ono’s compound towards clinical development.
For its research services, Evotec will receive research funding and milestone payments, the Hamburg-based company said.
“We anticipate the collaboration will result in identifying a novel drug candidate with a high potential,” Ono’s Managing Director, Kazuhito Kawabata, said in a statement.
Specific financial terms of the agreement were not released.
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Researchers find candidates for new HIV drugs
Last Updated on Tuesday, 1 December 2009 11:26 Written by Editor Tuesday, 1 December 2009 11:26
While studying an HIV protein that plays an essential role in AIDS progression, researchers at the University of Pittsburgh School of Medicine have discovered compounds that show promise as novel treatments for the disease.
HIV drug discovery efforts have met with little success in finding compounds that interact with an important HIV virulence factor, called Nef, because it lacks biochemical activity that can be directly measured, explained Thomas E. Smithgall, Ph.D., William S. McEllroy Professor and Chair, Department of Microbiology and Molecular Genetics, and senior author of the paper, which was published last week in the early, online version of ACS Chemical Biology.
To get around that problem, Dr. Smithgall’s team developed an assay to measure Nef function indirectly by coupling it to another protein, called Hck, which Nef activates in HIV-infected cells. Because Hck activity can be easily measured, the investigators were able to use it as a reporter for Nef activity in an automated high-throughput screening process. In collaboration with the University of Pittsburgh Drug Discovery Institute, they screened a library of 10,000 chemical compounds against the coupled proteins to see which ones influenced Nef-induced activation of Hck.
After further testing, they confirmed that three compounds inhibited the activity of the Nef-Hck complex and, more importantly, all of them also interfered with HIV replication. One compound was so effective that it suppressed HIV replication to undetectable levels in cell culture experiments.
“So we now have a way to rapidly and efficiently screen for inhibitors of Nef signaling through Hck,” Dr. Smithgall said. “But the surprise was that some of those inhibitors also showed strong antiviral activity in cell culture models.”
There is evidence that people infected with HIV variants that have mutations in the Nef gene take substantially longer to develop disease symptoms or AIDS, he said. In animal models, disrupting the production of Nef from the virus or its interaction with Hck also delays or prevents disease symptoms. The next challenge for the researchers will be to determine whether these compounds also interfere with progression of AIDS-like disease in animal models by blocking Nef function.
“Most current therapies for HIV infection use drugs that interfere with the function of viral enzymes such as reverse transcriptase or with the interaction of the virus and the host cell,” Dr. Smithgall said. “Targeting Nef represents an entirely new approach that could be useful to deal with issues such as drug-resistant HIV strains, and may slow the progression to AIDS.”
He added that Nef is just one of several so-called “accessory proteins” encoded by HIV which are important virulence factors in AIDS. Inhibitory compounds against some of the others might be revealed using a similar coupled protein approach for high throughput screening.
Source: labspaces.net
Posted under Compound Screening, Discoveries, Innovations and Patents, Drug Development, HIV Research, HT Screening, New Drugs, Press Releases | Comments Off
SwitchGear Genomics Launches Novel High-Throughput Screening Products for Profiling Biological Pathway Regulation
Last Updated on Tuesday, 1 December 2009 11:21 Written by Editor Tuesday, 1 December 2009 11:21
Collections of Experimentally-Validated Human Promoter Reporter Vectors Empower Researchers to Measure Transcriptional Regulation Across Genes in a Variety of Different Biological Pathways MENLO PARK, Calif.--(Business Wire)-- SwitchGear Genomics, Inc., a leading provider of products for studying regulatory elements in the human genome, today announced the availability of the first cost-effective, high-throughput research tools for screening transcriptional activation and repression in a number of key biological pathways. The new SwitchGear pathway sets utilize experimentally-validated luciferase reporter vectors to accurately quantify human promoter activity from complete sets of genes associated with inflammation, cholesterol biosynthesis, oncology, vascular biology, nuclear hormone receptor signaling, and many other important biological pathways. "The SwitchGear panels of human promoter targets was selected from our genome-wide reporter collection of promoters using motif analysis and published functional genomic data sets," said Shelley Force Aldred, co-founder and President of SwitchGear Genomics, Inc. "We then performed pathway-specific inductions to create an activity profile across the set of constructs. We offer complete pathway profiling sets of constructs in high-throughput plate formats that empower researchers to efficiently profile the effects of many compounds and conditions." In addition, the company provides a smaller subset of "key responder" promoter constructs that showed a strong induction response in the experiments and which may be used as biomarkers in primary screening applications. Researchers at the National Institutes of Health screened over 1400 compounds to test hypoxia pathway stimulation and published the results in an article entitled "Identification of Chemical Compounds that Induce HIF-1alpha Activity." The screening process, including the use of the SwitchGear Genomics hypoxia set of promoter reporter assays, differentiated between 3 hypoxia mimetics and 2 other compounds that triggered the pathway independent of HIF-1alpha, a result important for effective compound screening. In addition to the hypoxia (HIF1a) pathway products, SwitchGear offers the following reporter assay profiling sets in both plate format and biomarker subsets: CREB, NF-kB, heat shock (HSF), p53, STAT, serum response factor (SRF), and cholesterol biosynthesis (SREBP). In addition, the company offers nuclear receptor sets for estrogen receptor, androgen receptor, and glucocorticoid receptor pathways. About SwitchGear Genomics, Inc.: SwitchGear Genomics, Inc. is a leading provider of products for studying regulatory elements in the human genome. The company has developed a comprehensive approach to generate new insights into gene regulatory networks and allow researchers to efficiently screen entire pathways in living cells. SwitchGear was founded in March 2005 by Dr. Richard Myers, Dr. Nathan Trinklein and Dr. Shelley Force Aldred from Stanford University. For more information about SwitchGear, please visit the company's website at http://www.switchgeargenomics.com.
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Broad Institute Puts Genedata’s Screener to Work for High-Throughput Screening Data Analysis
Last Updated on Monday, 30 November 2009 01:59 Written by Editor Monday, 30 November 2009 01:59
Genedata announced today that the Broad Institute is using its Screener software platform to manage and analyze high-throughput screening data as part of its participation in the National Institutes of Health’s Molecular Libraries Roadmap Initiative.
“We’re screening upwards of 50 assays and analyzing more than 20 million wells of screening data” per year, Dave DeCaprio, associate director of the Chemical Biology Platform at the Broad Institute, said. He told BioInform that the Broad has been using the software since May, and that has reduced the time for data analysis from a “few weeks” to hours.
DeCaprio said that he and his colleagues chose Screener because it “is able to analyze a lot of data,” but only needs a “nominally powered-server.” The software sits on the Broad’s local server farm and “the client component gets served out through a web browser.”
Since follow-up biology and chemistry is “extremely expensive” after high-throughput screening, it’s critical to be able to query data from the screens as quickly as possible in order to identify potential problems, he said.
“One of the things we wanted was the ability to have strong analytics on the data, so we could automatically detect problems,” while at the same time offering an interactive feature so he and his team can intervene while looking at the data, he said. “They could quickly jump into the data and see what it really looked like.”
DeCaprio and his team validated the software against “some existing systems we had and existing approaches” from the “top five vendors,” but he declined to offer vendor or software names.
Open source tools he did not wish to name were also part of the evaluation but he said the team didn’t see anything that would support “what we wanted to do in terms of the visualization capabilities.”
“There are great algorithms you can get,” DeCaprio said. “The combination of algorithms, visualization, and manual curation is, I think, incredibly essential to getting high quality data out. That’s not something we saw anywhere else.”
“The key thing for us was the ability to integrate manual curation of the data with the algorithms,” DeCaprio said.
A user can look at all the results from a six-week screen, “visually spot some problems and make corrections, and the algorithms would adjust the data analysis based on that,” he said.
Small Molecule Test Drive
DeCaprio is responsible for the Broad’s infrastructure for small-molecule screening and further development, which includes informatics, compound management, data analysis, and analytical chemistry, as well as the procedural side of the work, he said.
The platform is a “public screening center,” he said. The Broad Institute’s Probe Development Center is one of nine centers funded under the NIH’s Molecular Libraries Probe Production Centers Network, which kicked off last year with $70 million in funding over four years to accelerate the pace at which small molecule probes are developed.
Typical customers for the Broad’s chemical biology platform are scientists who have identified a potential molecule and would like to put it “in front of 350,000 compounds,” he told BioInform.
These scientists can first apply to the NIH, and if accepted, the Broad Institute takes in their assay, runs the screen, and does follow-up chemistry “to basically get them to a chemical probe” DeCaprio said.
In addition to the NIH application process, scientists can apply directly to the Broad and work on a fee-for service basis.
Some of the Broad’s clients include Princeton University’s Bonnie Bassler and Stanford University’s Jerry Crabtree, several research teams at the Dana-Farber Cancer Institute, and Massachusetts General Hospital.
Customers for the service are generally academics, DeCaprio said.
“We’re happy to work with anybody, but one of our goals is to make all the data publicly available,” he said. “Everything we do goes into the PubChem database,” which most pharmaceutical firms and biotechs would rather avoid, he said.
Capture It
The intake process at the Broad involves first replicating an outside researcher’s work and then running the screen. Unlike traditional labs, he said, The Broad puts robotics to work on a protocol and all steps and changes are captured in a CambridgeSoft electronic lab notebook system that is made available to the researchers who requested the screen, he said.
“We use that to track all the interactions so that all of the metadata about what’s going on [in] experiments” is captured, DeCaprio said.
Primary data for 325,000 compounds comes off a detection instrument and is processed using Screener to “do QC, correct for controls, [and] normalize the data,” he said. Part of the process is automated but some decisions are manual since it is a “complicated process with a lot of points of failure.”
Some data management steps include a scientist manually marking a screen result as “invalid” or ” what I expected to see,” he said. Screener enables this level of interactivity, which is a feature that DeCaprio appreciates.
After a review of the first set of results with the collaborator, the team can decide which follow-on steps are necessary.
The Broad team is using Screener’s Assay Analyzer module, which visualizes the “raw well-level data.” It captures data from plate readers and processes them according to “predefined business rules.”
A separate module, Condoseo, plots the data on dose-response curves, to give scientists information such as IC50 numbers, which sheds light on the potential effectiveness of a compound.
All of the data stays at the Broad Institute in an “open data-sharing environment.” Participating scientists contribute their biology and findings and sign a data-sharing agreement. “They get access to everybody else’s data,” he said.
“Eventually it all goes public, after a year,” he said. In the first year, it is an environment “where people can access it privately.”
While high-throughput screening data sets are “not huge” — especially compared to ” the next-generation sequencing problems we have,” DeCaprio said that HTS data has its own challenges.
For example, results are “heavily dependent” on the conditions and the context of an assay, DeCaprio said, so he and his colleagues focus on the metadata, the “richness of the data,” associated with the results.
“In the small-molecule space you absolutely have to have” metadata, or the results can end up being “useless.” In a separate project with CambridgeSoft he is working on metadata management “to really try and open up that metadata and make it far more searchable,” he said.
The metadata is captured in the ELN, which is integrated with Screener, he explained, adding that the Broad team used Screener’s APIs to build the integration.
Source: Genomeweb
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Scientists get closer to making safe patient-specific stem cells
Last Updated on Monday, 30 November 2009 01:55 Written by Editor Monday, 30 November 2009 01:55
Scientists are a big step closer to their long-term of goal of creating patient-specific stem cells that are safe to use and don’t require the destruction of embryos.
Induced pluripotent stem cells – also known as iPS cells – are all the rage in the nascent field of regenerative medicine. Like embryonic stem cells, they have the potential to become any type of cell in the body and could be used to grow replacement parts, such as insulin-producing beta cells for diabetes patients or nerve cells for repairing spinal cord injuries.
Even better, they can be made by reprogramming skin or other cells from the patients who need them. That not only eliminates the need to use embryos, it ensures that the replacement tissues made from iPS cells are genetically matched to patients and won’t be rejected by the body’s immune system.
But there’s still a big catch: In order to rewind adult cells to a pluripotent state, researchers have to turn on a set of dormant genes that have the potential to cause tumors. So do the viruses they use to activate those genes.
So researchers have been looking for ways around this problem. One approach is to snip out the genes and viruses once the reprogramming is complete. Another is to use DNA sequences called transposons in place of viruses, then delete the transposons after they’re no longer needed. One group of researchers has even used genetic engineering to modify the key genes so that they can enter the skin cells without requiring viruses or transposons.
But many scientists think the safest approach is to replace the genes altogether with so-called small molecules. In a study published online today in the journal Cell Stem Cell, researchers from the Harvard Stem Cell Institute report that a single compound they dubbed RepSox can replace two of the four key reprogramming genes.
“We’re halfway home, and remarkably we got halfway home with just one chemical,†senior author Kevin Eggan, a professor in Harvard’s department of stem cell and regenerative biology, said in a statement.
Eggan’s team identified RepSox by screening 200 compounds and waiting a couple of weeks to see which of them did the best job of transforming mouse cells into iPS cells in combination with three of the four reprogramming genes. The researchers were surprised to find that their compound not only replaced the gene Sox2 (hence the name RepSox), but also made the gene c-Myc obsolete.
Now the group will turn its attention to finding other small molecules that could replace the remaining genes – Oct4 and Klf4 – as well, “opening a route to purely chemical programming,†they write.
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Scripps Research scientists awarded $3.9 million grant to develop new compound screening platform
Last Updated on Tuesday, 20 October 2009 10:04 Written by Editor Tuesday, 20 October 2009 10:04
Bicoastal effort could help revolutionize the search for new therapies
La Jolla, CA, and Jupiter, FL, October 5, 2009 –A pair of scientists from The Scripps Research Institute, one on each coast, has been awarded a five-year $3.9 million grant from the National Institutes of Health (NIH) to develop a new technology to accelerate the search for new protein ligands – compounds that bind to proteins and alter their function.
Current screening technology, which is slow and expensive, has caused what the NIH calls a “major bottleneck” in the search for these basic tools that are key for the broader study of biological processes and that lay the groundwork for development of most drugs.
The grant, awarded as part of the NIH’s new Roadmap Transformative R01 Program, will be shared between the laboratories of Tom Kodadek, Ph.D., a professor in the Scripps Research Departments of Chemistry and Cancer Biology in Jupiter, Florida, and Benjamin Cravatt III, Ph.D., professor and chair of the Department of Chemical Physiology and member of The Skaggs Institute for Chemical Biology and Helen L. Dorris Child and Adolescent Neuro-Psychiatric Disorder Institute at Scripps Research in La Jolla, California.
“Ben and I are extremely pleased to win this highly competitive award and to be among the first selected for the new Transformative Grant program from the NIH,” Kodadek said. “This is a perfect example of the tremendous collaborative possibilities available within Scripps Research. We worked on the proposal together and the fact that we’re both part of the same national institution will make the work that much easier as we move ahead.”
Cravatt added, “This project is a good reflection of what those of us at Scripps Research in La Jolla and in Florida are trying to accomplish – fostering collaborative interaction and working on complimentary research projects. This will help cement the strong working relationship between our two campuses.”
The NIH Roadmap Transformative R01 (T-R01) Program awards were launched this year to support exceptionally innovative, high risk, original, and/or unconventional research projects that have the potential to create or overturn fundamental scientific paradigms.
“The appeal of the Pioneer, New Innovator, and now the T-R01 programs, is that investigators are encouraged to challenge the status quo with innovative ideas, while being given the necessary resources to test them,” said NIH Director Francis S. Collins, M.D., Ph.D. “The fact that we continue to receive such strong proposals for funding through the programs reflects the wealth of creative ideas in science today.”
Two Innovative Methods and a Cab Ride
The new Scripps Research project will combine two separate technologies from each laboratory – a peptoid library synthesis and screening platform developed in the Kodadek laboratory and an activity-based protein profiling system developed in the Cravatt laboratory.
Kodadek’s screening platform involves the creation of vast libraries of peptoids (peptoids are synthetic molecules that are similar to peptides, compounds that when joined together make up proteins) displayed on microscopic beads that are screened against fluorescently tagged proteins that light up after binding with a high affinity, highly selective ligand.
“Our screening technology simulates the cellular environment,” Kodadek said, “because the tagged proteins, which represent only a small fraction of the total, are mixed in with un-tagged competitors. There is a specificity filter built into the process from the beginning.”
The Cravatt Laboratory has pioneered the Activity-Based Protein Profiling technology, which allows scientists to identify protein classes based on their activity. The basic technology attaches a single label or probe to proteins from a particular subset of the proteome, which allows access to what are considered low abundance proteins and makes it ideal for massive parallel screening experiments. So far, Activity-Based Protein Profiling probes have been developed for more than a dozen distinct enzyme classes.
Cravatt’s technology makes it possible to target what he calls “interesting classes of proteins” but in a highly parallel fashion – hundreds of screens at a time of those multi- million member peptoid libraries. Although both scientists have known one another for some time, many of the details of the collaboration were worked out on a cab ride from England’s Heathrow airport to London last summer.
“Tom and I had an editorial board meeting in London, and shared a cab from the airport,” Cravatt said. “The fact that Tom had recently joined Scripps Florida helped get us energized about the project.”
“It’s true,” Kodadek added. “The ideas behind the grant proposal just popped out of that ride.”
A Transformational Marriage
The combination of the Kodadek and Cravatt advanced technologies will allow the screening of massive peptoid libraries (1-10 million synthetic compounds) in parallel fashion, a novel strategy that the scientists predict will increase the rate of ligand discovery by several hundred times over current methods.
“The gist of our proposal is quite simply marrying these two beautifully worked out technologies,” Kodadek said. “We have a good track record on both sides, plus we’re building off these innovative platforms, so if this works, and I’m certain it will, it will definitely be transformational.”
That transformation, when it comes, should result in more lead drug candidates, Kodadek said, because while the scientists’ success rate has been lower than those using current high throughput screening technology, the quality of the ligands identified has been significantly better. Some of this is due to the fact that simple synthetic compounds like peptoids have many advantages over other ligands such as antibodies. They can be modified easily for attachment to surfaces and can be produced in relatively large amounts at lower cost and rather quickly – a multi-million member peptoid library, for example, can be created in around three days.
“The way most science works today,” Cravatt said, “is that researchers tend to huddle around those areas where there are tools available. By combining our technologies, we will have a streamlined, unbiased way to identify high quality protein ligands and that will give us access to a large part of the proteome that others can’t study right now because the current technology is inadequate.”
About The Scripps Research Institute
The Scripps Research Institute is one of the world’s largest independent, non-profit biomedical research organizations, at the forefront of basic biomedical science that seeks to comprehend the most fundamental processes of life. Scripps Research is internationally recognized for its discoveries in immunology, molecular and cellular biology, chemistry, neurosciences, autoimmune, cardiovascular, and infectious diseases, and synthetic vaccine development. Established in its current configuration in 1961, it employs approximately 3,000 scientists, postdoctoral fellows, scientific and other technicians, doctoral degree graduate students, and administrative and technical support personnel. Scripps Research is headquartered in La Jolla, California. It also includes Scripps Florida, whose researchers focus on basic biomedical science, drug discovery, and technology development. Scripps Florida is located in Jupiter, Florida.
Source: eurekalert.org
Posted under Compound Screening, Discoveries, Innovations and Patents, Grants and Awards, Industry News, Press Releases | Comments Off
MALDI-Based Method May Reduce Cost of Rx-Screening Assays, Speed Drug Development
Last Updated on Tuesday, 20 October 2009 09:44 Written by Editor Tuesday, 20 October 2009 09:44
This story originally ran on Oct. 6.
By Tony Fong
Researchers from the University of Cincinnati and MDS Analytical Technologies have used mass spectrometry to develop a high-throughput screening method for drug discovery they say can be more precise and cost-effective than existing techniques.
The technique is based on a MALDI triple-quadrupole platform and exploits the selective multiple-reaction monitoring transition features of the platform. By doing so, the new method is able to lower the cost of high-throughput screening for drug compounds to pennies per well from as much as $1 per well currently, Ken Greis, associate professor of cancer and cell biology and director of proteomics and mass spectrometry at the University of Cincinnati College of Medicine, told ProteoMonitor recently.
A study detailing the method was published Sep. 15 in the online edition of Rapid Communications in Mass Spectrometry.
In the paper, Greis and his co-authors said that drug discovery typically begins with a validated target enzyme “with the initial goal of finding appropriate molecular scaffolds with inhibitory activity via high-throughput screening.” The scaffolds are then subsequently used for lead compound optimization, and “ideally for the development of a safe and effective therapeutic compound.”
The most common methods of high-throughput screening have been fluorescence- and chemiluminescence-based approaches. Such approaches, Greis said, have been “very successful” because the same reagents can be used for many different enzymes.
But that same characteristic also creates a risk for interference.
“When one’s evaluating a compound repository for inhibitors, you often have a series of compounds that will fluoresce themselves,” Greis said. “If they fluoresce, they’re going to give you a false signal. Alternatively … there are compounds that inhibit the fluorescent properties, or what’s called quenching fluorescence, [that] also give false read-outs.”
Another problem is in the way the assays get generalized so that the reagents work for a wide range of enzymes. Such assays are called coupled assays: “You have a product being formed from your enzyme reaction but that’s not what actually triggers the fluorescence,” Greis said. “That product gets converted to another enzyme to another product through another enzyme to another product that then can be fluoresced.â€
This series of enzyme step, or coupled assays, ultimately results in a read-out. “The problem is any compound that interferes with any of those steps along the way also gives you false read-outs,” which tend to be false positives, he added.
But by using mass spectrometry to measure enzyme activity, Greis and his colleagues are able to get a direct read-out, “so a mass spectrometer effectively can give you a quantitation and a mass of a compound.”
By taking a ratio of the substrate being converted to a product — the essence of an enzyme assay, Greis said — and measuring that directly on a mass spec, there is no interference either from quenching or auto-fluorescence.
“And what we’ve found thus far is we’ve not seen any false positive read-outs. If we get a compound that shows that it’s active, even in single-point assays, it’s been demonstrated that it’s a dose-dependent inhibitor.”
And because the method uses native peptides or small-molecule substrates, the method can be done for “at most, pennies per sample well,” Greis said. By comparison, fluorescent and chemiluminescence reagents cost between 50 cents to $1 per well.
“So if you run a million compounds, you can run up a half-million dollars of reagents costs right away, whereas the label-free read-out is going to cost you maybe a couple thousand dollars for the reagents,” he said. “That’s a mass spec advantage.”
A prior study by researchers in China had demonstrated the utility of a MALDI-Fourier transform mass spectrometer for high-throughput screening of small-molecule substrate/product conversion.
The researchers chose AChE because of its long history of enzyme assay development, including colorimetric assays, pH-change assays, and most recently aggregation-induced fluorescence assays and mass-spec assays.
Speed is of the Essence
They also chose a MALDI platform, rather than an electrospray platform, because of the higher speed that can be achieved on the MALDI. Most enzymatic reactions contain salts that can interfere with mass spectrometry. An ESI platform requires a desalting step, which limits the throughput to five to 10 seconds per sample. A MALDI-based approach skips the desalting step, however, because the technology is less sensitive to salts.
“Essentially all that we do is run the enzyme reaction on a 384-well format,” Greis said. “We transfer all at once into a matrix plate mix and onto our MALDI target plate.”
Because there are no cleanup steps on the MALDI triple-quad, samples can be scanned at up to three samples per second, he said.
Greis acknowledged that the MALDI technology, especially the MALDI triple-quad, is not a popular tool for drug discovery. In his opinion, that’s because drug-discovery researchers were trained on electrospray mass specs and are comfortable with them.
“To then move them into a MALDI platform that they don’t understand, they’ve got a bias that it can’t be quantitative, and all these sorts of things from earlier studies using MALDI-based approaches that have been demonstrated time and time again to not be true anymore — I think there’s a cultural thing,” he said.
A criticism of a MALDI approach is that while it works well for peptide substrate screening, it doesn’t for small-molecule substrate products because of matrix interference in the low mass range.
“And we show very directly … that by taking advantage of the transition,” a chemical fragmentation that is diagnostic of a substrate or product “that one can do in a triple-quad, that matrix interference completely goes away,” Greis said.
The researchers tested their method by screening a library of 1,008 structurally diverse compounds across 384-well microtiter plates as an example of a single-dose primary screen, and reported that all known AChE inhibitors resulted in complete inhibition of enzyme activity, as expected. The hits were then validated “by demonstrating concentration-dependent inhibition and the rank order of inhibitory potency in hit follow-up assays,” they said in their study.
The technique they’ve developed can also be used on a simple MALDI instrument, though it works best for peptide substrate enzymes. With low molecular-weight enzymes, sensitivity can be an order of magnitude lower on a simple MALDI “because you’d have to be using enough enzyme substrate product to see your substrate products down in those low mass ranges in amongst all of the matrix peaks,” Greis said.
Also, Greis said there will be enzymes — such as fatty acids and long-chain hydrocarbons —that will not be amenable to a MALDI-based approach.
“The fact of the matter is that any mass spectrometry-based technique is only as good as the molecule that it’s trying to evaluate,” he said. “We have to be able to ionize the substrate and/or the product to be able to measure and quantify it.”
In ongoing work, he and his team members are developing multiplex assays. The typical screening approach is to take a target enzyme and pass the whole repository across it to look for inhibitors, and then validate the inhibitors. The next therapeutic target is then set up and the process is repeated.
With a mass spec-based approach, “as long as your enzymes reactions are compatible … you can run multiple enzymes in one pot and pass your repository against it once and get hits for all those different enzymes,” Greis said.
In conferences, Greis and his colleagues have presented proof-of-concept studies that show that “this in fact works quite well using a kinase and acetylcholinesterase or a kinase with a protease all in the same part,” he said. “We’ve shown that we can get selective inhibitors for each of them individually without interference in the multiplex format.”
Source: genomeweb.com
Posted under Compound Screening, HT Screening, Industry News, Press Releases | Comments Off
Trophos launches the new generation of its fast plate imaging instrument, the Plate RUNNER HD
Last Updated on Thursday, 22 October 2009 11:09 Written by Editor Monday, 12 October 2009 03:31
Marseille, FRANCE, 2009 September 29 – Trophos SA, a clinical stage pharmaceutical company developing innovative therapeutics for indications with under-served needs in neurology and cardiology, announced today the launch of the new generation of its fluorescence fast plate imaging system, the Plate RUNNER HD(R).
Trophos originally developed the Plate RUNNER HD(R) to speed up its own screening campaigns for drug discovery and development. The company has now implemented an improved version which offers much better resolution and allows imaging of the 7mm well of the standard 96 plate in matrices of up to 8192 x 8192 pixels. This positions it as the superior alternative to the traditional automated microscope in a wider area of cellular and small organism imaging applications such as neurite outgrowth or C.elegans screens. The Plate RUNNER HD(R) is protected by patents granted in the USA and Europe.
“The Plate RUNNER HD(R) has now reached both a performance and an operational level that allows it to be widely used in various imaging platforms,” said Damian Marron, CEO of Trophos. “We have optimized it by regular use in our screening and research platforms. Use in other labs such as the Neuronal Cell Biology & Pathology research unit directed by Dr Christian Neri at the Inserm Psychiatry and Neurosciences Centre (Paris, France) and the Laboratory of Motor Neuron Biology directed by Dr Alvaro G. Estévez at the Burke Medical Research Institute (NY, USA) has confirmed its performance.” (For further information, see http://www.broca.inserm.fr and http://www.burke.org).
The Plate RUNNER HD(R) fills the gaps between the low sensitivity/low resolution/cell consuming fluorescence plate readers, the high sensitivity but non-imaging flow cytometer, and the slow, poorly automated and small field fluorescence microscope. It does this by allowing whole well/full plate imaging in just 2 minutes 40 seconds for 96 images of 1024 x 1024 pixels, 8 minutes at 4096 x 4096 resolution and 25 minutes at the extremely powerful 8192 x 8192 resolution. The maximum resolution gives details as fine as 1 micron, which covers the vast majority of cellular imaging applications.
“The instrument is not only the source of our pipeline – nearly 1 million compounds screened in eight years – but also an everyday research tool that does in two minutes a job that usually requires a week manually,” said Pierre Delaage, Head of Development of Trophos Instruments. “Using exclusively open data formats, the Plate RUNNER HD(R) integrates smartly and smoothly with all existing imaging and data processing environments with no hidden extra costs. It is also a “green product” using at least ten times less electrical power for illumination than its competitors. It has the important advantage of having a lower price and TCO than the traditional microscope-based instruments.”
Dr Christian Neri, Research Director at the Psychiatry and Neuroscience Center of Inserm in Paris, stated “We have been using the Trophos screening system since its first commercial version; it allowed us to simply and rapidly automate our screening protocols on C.elegans worms and on cells. To move further in C.elegans screens we needed a far higher resolution without sacrificing light, sensitivity and field. We have now seen that Trophos’ latest instrument meets perfectly our very stringent requirements.”
“We continue to directly sell and support the Plate RUNNER HD(R) and are now looking to set up a commercial and industrial partnership to address more widely the global market opportunity,” added Marron. “The Plate RUNNER HD(R) offers unique performance and can be a great fit for imaging solutions manufacturers and vendors, to exploit opportunities for example in the area of small organism or angiogenesis screens as well as cellular screens.”
About the Plate RUNNER HD(R): Developed by Trophos since 1999 initially for its own HTS platform, The Plate RUNNER HD(R) is a rapid fluorescence 96-plate imager giving full 7mm well images in single snapshots (ie no mosaic reconstruction), embedding three commonly used wavelength illumination lamps using fast-switching LED technology, thus leading to more homogeneous light, very long life time (100 000 hours against 5000 hours for xenon or mercury lamps), higher speed and dramatically reduced maintenance costs. Images are given in standard 1024 x 1024 definition but also in higher 2048 x 2048, 4096 x 4096 and even 8192 x 8192 pixels definition, leading to details of about 1 micron in size.
Compared to classic automated microscopes, The Plate RUNNER HD(R) is much more simple, integrated, automated, robust and fast; it has no oculars (eyepieces), no huge frame, no complicated auto focus system, no complicated settings, no complicated software, and does not need any cooling/heating delay between illumination sessions nor painful recalibration between sessions. Built on modular, open and interoperable design and data formats, it is compatible with any existing image processing platform. Training for common tasks is achieved in less than 1 hour, and there is no need of high skilled people to install or set it up.
Driven by Trophos proprietary software running on standard low cost PCs and using advanced real time programming techniques coming from the industry, the device reliably acquires 96 images at 1024 x 1024 definition in just 2 minutes 40 seconds (for a standard 488nm excitation and 200ms exposure time), 8mn for 4096 x 4096 images and 25mn for 8192 x 8192 images.
The device is patent protected in France, Europe and USA. Patent is pending in Canada and Japan. Detailed specifications are available here (http://www.trophos.com/download/pr_datasheet.pdf).
About Trophos: http://www.trophos.com Trophos is a clinical stage pharmaceutical company developing innovative therapeutics from discovery to clinical validation for indications with under-served needs in neurology and cardiology. The Company has a novel and proprietary cholesterol-oxime based chemistry platform generating a pipeline of drug candidates, with the lead product, olesoxime (TRO19622), in phase II clinical trials and a second product, TRO40303, planned to enter the clinic in 2010. Trophos’ mitochondrial pore modulator compounds enhance the function and survival of stressed cells via modulation of dysfunctional mitochondria through interactions at the permeability transition pore (mPTP). Recently published clinical studies support the therapeutic rationale for mitochondria targeted drugs in neurology (Alzheimer’s disease) and cardiology (ischemia-reperfusion injury), which Trophos is uniquely placed to exploit.
Trophos has not only invested in science but also in technology such as the Plate RUNNER HD(R), which is a key reason why Trophos was able to bring products into phase II clinical trial after only 8 years, instead of the 12-15 or even 20 years commonly observed”.
For further information, please contact:
Andrew Lloyd & Associates Andrew Lloyd / Neil Hunter Tel: +44 1273 675100 allo@ala.com
Source: medadnews.com
Posted under Compound Libraries, Compound Screening, Industry News, Press Releases | Comments Off
GENFIT identifies compounds which modulate clock genes for the treatment of cardiometabolic disease and CNS disorders
Last Updated on Monday, 12 October 2009 02:29 Written by Editor Monday, 12 October 2009 02:29
Lille (France), Cambridge (Massachusetts, United States), September 28, 2009 – GENFIT (Alternext: ALGFT; ISIN: FR0004163111), a biopharmaceutical company at the forefront of drug discovery and development, focusing on the early diagnosis and preventive treatment of cardiometabolic and neurodegenerative diseases, today announces the successful identification of Hit compounds for an orphan nuclear receptor which plays a key role in the regulation of circadian cycle in different organs. These Hits were identified through the screening of chemical libraries performed at GENFIT facilities in Lille.
In humans, many aspects of behavior and physiology are coordinated by an endogenous circadian rhythm (circa diem, meaning approximately one day) that is generated by an internal clock system which synchronizes daily variations in gene expression to rhythms such as sleep and wake alternance, variations in body temperature, blood pressure, heart rate, as well as cognition, attention and mood.
A large body of evidence from both human and animal studies now points to a relationship between circadian disorders and altered metabolic response, suggesting that circadian and metabolic regulatory networks are tightly interconnected.
As a consequence, misalignment of the internal timing system versus environmental stimuli, such as day/night cues, as experienced during jetlag or shift work, may result in dysregulation of physiological cycles of fuel utilization and energy storage, and has been associated with increased risk to develop obesity, type 2 diabetes, hyperlipidemia, high blood pressure and cardiovascular disease. As well, modulating, resetting and stabilization of central circadian rhythms have been proposed as therapeutic strategies for certain CNS disorders.
“This is an important milestone in our drug discovery programs in the field of cardiometabolic disease and CNS disorders”, says Dean Hum (CSO of Genfit). “Deorphanisation of this nuclear receptor further demonstrates our expertise in this class of therapeutic targets, and provides novel series of compounds to address the focus therapeutic areas of Genfit with a very innovative approach via modulation of clock genes and the circadian rhythm”.
About GENFIT:
GENFIT is a biopharmaceutical company focused on the Discovery and Development of drug candidates in strategic therapeutic fields linked to cardiometabolic and neurodegenerative disorders (prediabetes/diabetes, atherosclerosis, dyslipidemia, obesity, Alzheimer’s…). GENFIT uses a multi-pronged approach based on early diagnosis, preventive solutions, and therapeutic treatments to address these major public health concerns and their unmet medical needs. GENFIT’s proprietary research programs and its partnerships with leading pharmaceutical companies, including Sanofi-Aventis, Solvay Group, Pierre Fabre, and Servier, have resulted in the creation of a rich and diversified pipeline of drug candidates at different stages of development. GENFIT’s lead proprietary compound, GFT505, is currently in Phase II and two other compounds, in partnership with Sanofi-Aventis (AVE0897) and SOLVAY (SLV341), are in the advanced stages of Phase I.
With facilities in Lille, France, and Cambridge, MA (USA), the Company has about 130 employees, including over 100 scientists. GENFIT is a public company listed on the Alternext trading market by Euronext(TM) Paris (Alternext: ALGFT; ISIN: FR0004163111). www.genfit.com
Contacts:
GENFIT
Jean-Fran̤ois Mouney РChairman of the Management Board
+33 (0)3 20 16 40 00
Milestones – Press Relations
Bruno Arabian
+33 (0)1 75 44 87 40 / +33 (0)6 87 88 47 26 – barabian@milestones.fr
Copyright Hugin
The appendixes relating to the press release are available on:
http://www.hugingroup.com/documents_ir/PJ/CO/2009/158601_88_7960_20090928-PR-GENFIT.pdf
This announcement is originally distributed by Hugin. The issuer is solely responsible for the content of this announcement.
Source: euronext.com
Posted under Compound Screening, Discoveries, Innovations and Patents, Industry News, Press Releases | Comments Off
DiscoveryBioMed, Inc. Awarded Phase 2 SBIR Grant by the NIH to Discover Hypertension and Cystic Fibrosis (CF) Drugs
Last Updated on Monday, 12 October 2009 12:50 Written by Editor Monday, 12 October 2009 12:50
BIRMINGHAM, Ala.–(BUSINESS WIRE)–DiscoveryBioMed, Inc. (DBM) today announced that it has been awarded a $750,000 Small Business Innovations Research (SBIR) Phase 2 grant by the National Institutes of Health (NIH) to continue the research into the discovery and development of small molecules to alleviate multiple chronic human diseases including cystic fibrosis (CF), hypertension and chronic kidney diseases with hypertension.
“We are proud to have been awarded this grant and to have our technology again recognized and validated by the NIH,†said Dr. Erik Schwiebert, Chief Executive Officer of DiscoveryBioMed. “With our academic partners at the University of Alabama at Birmingham and at Johns Hopkins University School of Medicine, we stand ready to test lead compounds for safety and efficacy in both CF and hypertensive animal models.â€
DBM has adapted a known electrical bioassay method to be high-throughput screening friendly, a necessary solution to bring the bioassay to the molecular target endogenous to the apical cell membrane of polarized renal and respiratory epithelia. The molecular target in play for this drug discovery program is an epithelial ion channel that is the rate-limiting step for the handling of salt in the distal portions of the kidney and in the respiratory tract. When over-active, this sodium channel can cause dehydration of the airways and too much salt in the blood, leading to high blood pressure.
“To successfully study this ion channel target, we had to bring the bioassay to the target where it is most comfortable, the apical membrane of a polarized epithelium simulated in in vitro 3D culture,†continued Dr. Schwiebert. “Researchers refer to this target as ‘twitchy’ since it does not behave the same in other experimental systems. It also depends upon factors produced by the epithelium itself to maintain proper activity. DBM brought the assay to the target and remains true to the principle that the target should be endogenous to a human or mammalian epithelial cell system to empower the most biologically-relevant drug discovery program. We believe screening on life-like human cell platforms is essential in development of drugs that ultimately will be provided to human patients.â€
Additionally, DiscoveryBioMed has a pair of closely related lead compounds in hand that it will use as a medicinal chemistry platform. Additional hit-to-lead compounds are emerging. At the end of Phase 2, DBM anticipates having pre-clinical animal data and, possibly, proof-of-concept efficacy data in animals and in humans to show to potential out-license partners.
About DiscoveryBioMed, Inc.
DiscoveryBioMed, Inc. is a life sciences and biotechnology company that engages in R&D and services work in cell engineering and production and cell-based drug discovery. The company is located within The Innovation Depot facility in Birmingham, Alabama. Using physiologically relevant cell culture models preferably derived from normal and diseased adult human cells and tissues, DBM focuses on finding therapeutic compounds for a variety of human diseases. It also applies this custom human cell-based approach to its “fee-for-service†support to researchers in allied areas and currently serves clients both locally in Alabama as well as in 11 other states in the US currently. For more information, visit the DBM website at www.discoverybiomed.com.
Source: Businesswire.com
Posted under Business and Investment, Clinical Trials, Collaborations, Compound Screening, Press Releases | Comments Off
Sirona Biochem Says SGLT Test Results Confirm Key ‘Breakthrough’
Last Updated on Friday, 9 October 2009 12:02 Written by Editor Friday, 9 October 2009 12:02
Sirona Biochem Corp. (TSX-V: SBM), an emerging biotech company focused on diabetes and obesity, says results of testing its unique SGLT inhibitor molecules demonstrate a key breakthrough milestone for Sirona Biochem.
Sirona Biochem CEO, Dr. Howard Verrico, said, “There are two vital steps in the early stage of drug testing: validation of concept i.e. a molecule is able to hit the desired target and secondly its in vivo effectiveness. This first round of testing has shown a key breakthrough milestone in the process of validating this concept.”
“The test results now mean we can proceed to find out whether the molecules are selective, safe and robust enough to have potential to be effective when they reach the parts of the body where the re-uptake of glucose needs to be limited.”
Dr. Bertrand Plouvier, Chief Scientist, said, “The results from the first round of screening are indeed very encouraging and Sirona Biochem will use the next following months to further study the molecules through specific assays to demonstrate their effectiveness and drug likeness.”
Dr. Verrico said management of sugar metabolism is a primary medical challenge associated with treating diabetes and obesity and that is why SGLT inhibitors show such promise in this regard. “At present SGLT2 inhibitors have demonstrated their ability to limit the re-uptake of glucose back into the blood stream from urine. However, they have been notoriously lacking in ability to resist being rapidly metabolized by the body, thus rendering them largely ineffective.
“What we have now done is show that our molecules, with their unique GlycoMim® technology, can inhibit the glucose transporter SGLT2. The next challenge, and an exciting one, is to show that our molecules are selective, safe and have the potential to have an increased efficacy compared to the current molecules undergoing clinical development.”
Sirona Biochem owns the worldwide product rights to a library of unique sodium glucose transporter (SGLT) inhibitors to treat diabetes and obesity. SGLT inhibitors, as previously stated, block the re-uptake of excess sugars from urine, which can then reduce high blood sugar towards normal levels.
Sirona Biochem has entered into a strategic partnership with TFChem, a drug discovery company based in Rouen, France. TFChem licenses its technology of fluorinated carbohydrate mimics: GlycoMim®, and products in development to biotech companies. This strategic partnership was completed by a detailed research and licence agreement signed on September 29, 2008.
23.6 million people, or 7.8% of the population of the United States, have diabetes. (February 2009 DACG.ORG)
Market Trends
In 2007, the prevention and treatment of diabetes and its complications was estimated to cost US$ 232 billion according to the International Diabetes Federation. By 2025, this is likely to increase to more than US$ 302.5 billion.
The diabetes drug market reached US$18 billion in 2005, and is expected to increase to $21-25 billion in 2011. With many new products yet to realise their full potential and the high incidence of T2DM expected in emerging markets, prospects for the sector look strong. Some of the fastest growing markets for diabetes are in emerging economies. India, China and Indonesia are in the top 5 for disease prevalence. The impact for both branded and generic drugs is considerable.
Furthermore, in recent years, obesity has become a major health problem for many post-industrial societies, so much so that in 2004, the United States Health and Human Services declared obesity to be a disease. The World Health Organization (WHO) projects that globally in 2005, 1.6 billion adults were overweight with at least 400 million adults obese. By 2015, approximately 2.3 billion adults will be overweight and 700 million will be obese. Obesity poses a major health risk because it greatly increases the risk of co-morbidities such as diabetes, cardiovascular diseases, arthritis, and cancer. Recognizing the potential for a new blockbuster market, major pharmaceutical companies have increasingly focused on obesity and its causes and, in the process, seeking to identify many potential targets and pathways that could be exploited to create novel therapies.
Sirona Biochem’s website is at: www.sironabiochem.com where we feature the most recent information about the company and its activities. Alternatively, investors are able to e-mail all questions and correspondence to info@sironabiochem.com where they can also request to be added to the investor e-mail list to receive all future press releases and updates or call John Dougherty, Corporate Development at 604-641-4466.
About Sirona Biochem
Sirona Biochem Corp. (TSX-V: SBM) is an emerging biotech company dedicated to the discovery and development of novel drug compounds. The current focus is on treatments for Type II diabetes and obesity. Sirona has entered into a license agreement with TFChem S.A.R.L., a drug discovery company based in Rouen, France. TFChem licenses its technology of fluorinated carbohydrate mimics: GlycoMim®, and products in development to biotech companies. The license agreement with TFChem provides for research and development of new compounds known as SGLT Inhibitors. SGLT inhibitors are a new and exciting class of compounds that have great promise and potential to treat both diabetes and obesity.
Mark Senner President and Director
Neither the TSX Venture Exchange nor its Regulation Services Provider (as that term is defined in the policies of the TSX Venture Exchange) accepts responsibility for the adequacy or accuracy of this release.
950-789 west pender street
vancouver, b.c., v6c 1h2
Direct: 604-641-4466
Fax: 604-608-5471
info@sironabiochem.com
Source: Marketwire
Posted under Cell Analysis, Cell-based Assays, Compound Libraries, Compound Screening, Discoveries, Innovations and Patents, Diversity Libraries, Drug Development, Drug-Like Compounds, Press Releases | Comments Off