Rule of Three considered harmful?

I should start this post by saying that I�ve never actually used the Rule of Three for fragment selection. Part of the reason for this is simply a matter of timing since I�d been designing fragment libraries before the Rule of Three came along. However, I believe that there are reasons that you need to take a very close look at the Rule of Three if you�re planning to build a fragment library strategy around it. The rule was introduced in late 2003:

�We carried out an analysis of a diverse set of fragment hits that were identified against a range of targets. The study indicated that such hits seem to obey, on average, a �Rule of Three�, in which molecular weight is < 300, the number of hydrogen bond donors is =3, the number of hydrogen bond acceptors is =3 and ClogP is =3. In addition, the results suggested NROT (=3) and PSA (=60) might also be useful criteria for fragment selection. These data imply that a �Rule of Three� could be useful when constructing fragment libraries for efficient lead discovery.�

My first criticism of the Rule of Three is that the authors do not say how they define hydrogen bond acceptors. I�ll illustrate this point with reference to the phenylhydantoin below which along with the accompanying properties was retrieved from eMolecules. As far as I�m concerned, this compound would have been perfectly acceptable for inclusion in a fragment library before the Rule of Three was published and the publication of the rule would not make change my mind. If, however, you asked me whether the compound complied with the Rule of Three, I�d have to admit that I simply don�t know. The number of hydrogen bond donors is not an issue because there is only one of these in the molecule. The number of acceptors is more problematic. I would only count the oxygen atoms in this molecule as acceptors and, since there are two of these, the molecule would be compliant with the Rule of Three. However the well-known Rule of Five treats all nitrogen and oxygen atoms as acceptors so if you use those criteria you�ll count a total of four acceptors and conclude that the compound is not compliant with the Rule of Three. This is not a problem for me because I don't use the Rule of Three but spare a thought for the person assembling a commercial fragment library.


My second criticism of the Rule of Three concerns how it was actually derived. The authors describe performing �an analysis of a diverse set of fragment hits� without actually saying anything about what this analysis entailed. If they were analysing hits from their own fragment screens then the characteristics of the hits will reflect the criteria by which compounds were selected for fragment screening. If they were sampling from a more extensive database of screening hits, I�d still want to know how the fragment hits were distinguished from the other hits.

My third criticism is as much about how cut offs get used as it is of the Rule of Three. There�s a diagram of a funnel that you often see in virtual screening reviews. We also use funnels (or filters as we prefer to call them) in screening library design and in fact this activity is not a whole lot different from working up a virtual screen. Typically we apply filters and sample (e.g. using molecular diversity criteria) from what makes it through. Note that I say �filters� rather than �a filter�. The Core and Layer (CaL) approach to library design has been described both in this blog and in a journal article. In CaL the filters used prioritise compounds get less restrictive as more compounds are added to the library. The reason for doing this is that it gives better control of chemical space coverage since it forces the selection of the smallest and least complex molecules first. A molecular diversity maximiser such as BigPicker, will tend to pick larger, more complex molecules because these tend to be more dissimilar to each other.

I am also prepared to accept compounds that have measured/calculated logP values in excess of 3 provided that the appropriate precautions (select ionisable compounds and/or use measured solubility values) have been taken to minimise the risk of poor solubility. You don�t want a whole library of compounds with logP values in excess of 4 but having some will increase the range of targets that you can nail. I am more concerned about the distribution of logP and molecular size in a library than I am with their maximum values and believe using multiple cut offs allows better control of these distributions.

You'll find plenty of material on the internet that deals with the Rule of Three although inconsistencies can be observed. It is not clear whether or not the Rule of Three includes the restrictions on NROT and PSA. As I read it in the original article, I don't think it does but I'm not sure and think it could have been made clearer. This webpage (accessed 11-Jan-2011) appears to suggest that Maybridge FBDD team think that the NROT and PSA criteria are included in the Rule of Three. However, another webpage (accessed 11-Jan-2011) seems to suggest that the FBDD team at Chembridge think otherwise. Cambridge Medchem Consulting (accessed 11-Jan-2011; I expect that this page will get updated once the error is discovered) appear to share the Chembridge view that the NROT and PSA criteria are not included in the Rule of Three although they use < instead of = when stating the Rule which makes a big difference when the number in question is 3. Yet another variation on the Rule of Three can be found in the BioScreening.net glossary (accessed 12-Jan-2011) in which the hydrogen bond criteria are stated as "number of H-bond donors and acceptors less than, or equal to 3", which could be taken to imply that the sum of donors and acceptors cannot exceed 3.

I should of course let you know where the title of this post comes from since I borrowed most of it from a computer science paper that is over forty years old. I can�t even claim originality for adapting the title of the earlier paper because my friends at OpenEye have beaten me to that as well. Furthermore my first two criticisms of the Rule of Three have already been made by other bloggers (accessed 11-Jan-2011).

I hope that this post will at least make people ask a few questions when presented with rules like these in the future. I'll also set up a discussion in the LinkedIn Medicinal Chemistry group which will facilitate posting of comments.

Literature cited

Congreve, Carr, Murray & Jhoti, A �Rule of Three� for fragment-based lead discovery? Drug Discov. Today 2003, 8, 876-877 | DOI

Lipinski, Lombardo, Dominy &Feeney, Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings. Adv. Drug Deliv. Rev. 1997, 23, 3-25 | DOI

Blomberg, Cosgrove, Kenny & Kolmodin, Design of compound libraries for fragment screening. JCAMD, 2009, 23, 513-525 | DOI

Dijkstra, go to statement considered harmful. Communications of the ACM, 1968, 11, 147-148 | DOI

BrazMedChem2010

It really was great to get back to Brazil. Early in November, I attended the 2010 Brazilian Symposium on Medicinal Chemistry that in Ouro Preto. The conference commemorated the work of Carlos Chagas who identified and fully characterized the disease that now bears his name. His work has been described in Wikipedia as unique in the history of medicine because he was the only researcher so far to describe completely a new infectious disease: its pathogen, vector (Triatominae), host, clinical manifestations and epidemiology. Chagas worked at and subsequently became the director of the medical research institute founded by Oswaldo Cruz. Although Chagas spent most of his working life in Rio de Janeiro, he was born a Mineiro and only left his home state for his university studies. Ouro Preto is one the must see attractions of Minas Gerais (and Brazil) and here are a couple of pictures to give those who were not there an idea of what they were missing.



Although the work of Carlos Chagas is worthy of celebration the current state of treatment of Chagas Disease is most definitely not. Like the better known African Sleeping Sickness, it is a trypanosomal disease and of minimal interest to Big Pharma. The drugs used for treatment have unpleasant side effects and once the disease enters the chronic phase they become a lot less effective. A number of lectures focus on the nature of the disease and I find these particularly interesting, although occasionally gruesome (I always appreciate these reminders of why I never considered pathology as a profession).

lvaro Romanha delivers an interesting keynote lecture in which he looks back over a long career in parasitology. He describes characterisation of the effects of a number of compounds (including one developed by my former employer) on the Chagas parasite. Lucio Freitas-Junior and Andrei Leit�o both talk about cell-based assays which can be used in target identification as well as lead discovery. I hope the folk doing natural product research are taking notes...

Barry Sharpless delivers an entertaining lecture on Click Chemistry and I also enjoy talks by Mike Gelb and Tom von Geldern since these both have a strong medicinal chemistry focus. The work described by Mike used Tipifarnib (1), a farnesyl transferase inhibitor as a starting point. This compound kills T. cruzi and its good pharmacokinetic properties reflect the fact that it had been in clinical development. It turns out that the compound actually inhibits the trypanosomal lanosterol 14-demethylase and this is the reason that it is able to kill the parasite. The optimized compound (2) shows efficacy against acute Chagas in mice and is sufficiently selective not to inhibit human lanosterol 14-demethylase or farnesyl transferase. The focus of Tom's lecture is African Sleeping Sickness although this is still highly relevant to Chagas Disease. He describes an interesting series of cyclic boronate esters (3), the mode of action of which is still uncertain. If you are going after Sleeping Sickness you are going to have to get your drug through the blood brain barrier. Tom describes some of the approaches that the team adopted to optimize pharmacokinetics and achieving good Central Nervous System (CNS) penetration. In the chronic phase of Chagas Disease the parasites take refuge in the cells of their host so the drug has an additional barrier to cross. I wonder how the presence of an intracellular parasite might stimulate expression of efflux transporters in a host cell? That would indeed be sneaky...


Cristiano Guimar�es presents analysis of relationships between permeability and physicochemical properties such as polarity and molecular size. However, I am more interested in what he has to say about re-scoring of docking poses. In particular, he notes that conformational entropy lost on binding tends to get over-estimated and has published some of this work. Enthalpy is the focus of the lecture by John Ladbury who describes how calorimetry can be used as a tool for understanding biomolecular interactions and an aid to drug design. The idea is that enthalpy changes associated with binding reflect the extent to which polar interactions form between the molecules in the complex. If you can exploit polar interactions to increase affinity then hopefully you�ll end up in a better place because polarity tends to be associated with better aqueous solubility, selectivity and metabolic stability. However, interpretation of the enthalpy and entropy changes associated with binding remains a challenging problem. I'd suggest taking a look at John's recent publication and recent discussion in the LinkedIn Medicinal Chemisry and Drug Discovery Group if you want to find out more.

That just about wraps up the technical part of this post and there is a page for links to talks. However, I did manage to get a few pictures including one of Carlos and another of his boss at the opening reception.



The following photos were taken at afternoon coffee on the last day of the conference. The first of these shows three of my friends from Rio and it was Daniel who did an excellent job introducing and chairing the molecular design session in which I spoke.



These pictures were taken towards the end of the conference. The paparazzo certainly knows how make the ladies from Porto smile and Carlos does look happy to be passing the baton to Vera who will be organising BrazMedChem2012.



Then it was time for dinner. Tom, Roberto and Ivan were staying in the annex and had to be be summoned. Tom looks a bit hungrier than the other two.



These folk are from FIOCRUZ in Belo Horizonte apart from Claudia (Federal University of Ouro Preto) and Malu who can't resist the photo opportunity. Andrei is looking quite intense in the next photo, Patricia less so.



These two photos are a couple of my favorites. I wonder if Barry is suggesting to John that the adiabatic stereoelectrostatic compressibility of the polarisability tensor may well be the the elusive Universal Efficiency Metric that he is searching for. Of course they could just be swapping fishing stories. One of the highlights of the dinner was Mike playing classical guitar and I was pleased that caipirinha consumption did not interfere with ability to operate a rather bulky digital SLR.



Literature cited

Guimar�es & Cardozo, MM-GB/SA Rescoring of Docking Poses in Structure-Based Lead Optimization. J. Chem. Inf. Model. 2008, 48, 958-970 DOI

Ladbury, Klebe & Freire Adding calorimetric data to decision making in lead discovery: a hot tip. Nat. Rev. Drug Discov. 2010, 9, 23-27 DOI

Molecular Interactions

Molecular interactions are an important part of the theoretical framework of modern drug discovery and studying them is a great way to increase your understanding of physicochemical principles of molecular design. One view of molecular design is as a process of tuning the interactions of molecules with the different environments in which they exist. Needless to say, knowledge of molecular interactions is particularly valuable in FBDD. Three Roche scientists have recently published �A Molecular Chemist�s Guide to Molecular Interactions� which should be of interest to anyone working in molecular design and other bloggers ( Derek | Joerg ) have already highlighted the article.

The authors cover plenty of ground and everybody should find their favourite molecular interactions discussed. Given the recent LinkedIn discussion on the value of measuring enthalpy and entropy changes associated with binding, I was pleased to see that the authors noted that interpretation of these quantities is typically difficult. The discussion of cooperativity was useful because we often assume that contributions of interactions to binding are additive.

One of my favorite interactions is halogen bonding and I am pleased to see it discussed in some detail. The halogens all confer a degree of hydrophobicity on a molecule and the heavier halogens (i.e. not fluorine) also exhibit an ability to interact with hydrogen bond acceptors that increases with atomic number. Although this class of interaction is sometimes thought to have been discovered in recent times, it�s actually been around a long time. I can remember learning, as a schoolboy in Port of Spain in the mid-1970s, about why iodine is more soluble in aqueous potassium iodide than in water. I developed this theme a bit more in a light-hearted survey of halogens at EuroCUP in 2008 which starred both Bismarck and a medical writer by the name of Bouchardat who was active in Paris when Pincess Victoria became Queen Victoria. I'm not sure if they ever caught the notorious Parisian dog-poisoner.

Something that I found disappointing was that there was not a lot of information about how much additional affinity you�re likely to get by making the different interactions. This should not be seen as a criticism of the authors who have carried out an impressive trawl of the literature. It�s just disappointing that the information is not available in the current literature base.

I�ve some comments to make on the discussion of hydrogen bonds. There is a widely accepted view that the maximum contribution to affinity that a hydrogen bond between a neutral donor and acceptor can make is just over a log unit and here�s what the Roche authors had to say on the subject:

�Hydrogen bonds always convey specificity to a recognition process but do not always add much binding free energy. Desolvation of the donor and the acceptor must occur for the hydrogen bond to form, such that the effects of hydration and hydrogen bond formation nearly cancel out�

I certainly agree that in some cases the contribution of hydrogen bonds to affinity will be minimal. However, the dataset from which that figure of just over a log unit was derived is actually quite small and not especially diverse in terms of donor-acceptor pairings. I believe that if you can form the hydrogen bond deep in a binding pocket then it can make more than the widely accepted maximum contribution to affinity. We recently published inhibition data which included the example of aza-substitution of a pyridine ring resulting in increases of potency of about two log units against Cathepsins S and L2. Some caution is required in interpreting these results because we didn�t have the relevant crystal structures and the inhibitors are racemic. However, I do believe that these results should make us question the prevailing view of the maximum contribution that a hydrogen bond between a neutral donor and acceptor can make to affinity.

I�ll say some things about the discussion of the hydrogen bonding of sulfonyl groups because it should get you thinking a bit. The authors state that:

�Only 30% of the sulfones and sulfonamides form hydrogen bonds. This raises the question of which type of interaction this functional group prefers.�

I�m not sure that I agree with the second sentence and would be interested to know how many of these sulfones and sulphonamides actually had the opportunity to accept a hydrogen bond. If no hydrogen bond donors are present in a molecule then you can�t really blame the sulfonyl oxygens for making contact with aliphatic carbon in the solid state since that's going to be a better option than getting in the way of the sulfonyl oxygens of a lattice neighbour. Even when a donor is present in the molecule, the favoured interaction may well be with a stronger acceptor than the sulfonyl oxygens.

The authors also took a look at the environments of sulfonyl groups in the PDB and here�s what they had to say.

�Notably, of the sulfonyl groups situated in a hydrophobic environment in the PDB, only 36% are found to interact simultaneously as a hydrogen bond acceptor but 79% of the hydrogen-bonded sulfonyl groups are found to interact simultaneously with a hydrophobic group. These findings clearly indicate a dual character of the weakly polar sulfonyl groups as a hydrogen bond acceptor and as a hydrophobic group.�

I simply don�t buy this idea of sulfonyl oxygens having a dual acceptor-hydrophobic character. Hydrophobicity is a statement of aquous solvation characteristics. It will be easier to place a weak acceptor in a hydrophobic environment than it will to place a strong acceptor there. However, if it�s an acceptor, it�ll still prefer the aqueous environment. Think about the consequences of one of these oxygens accepting a hydrogen bond. When an oxygen atom is hydrogen bonded its ability to accept a second hydrogen bond is likely to be reduced because the donor polarises the acceptor oxygen. Also the second donor will also experience repulsive secondary electrostatic interactions with the existing donor. Provided that the oxygen can still accept a hydrogen bond, it will not be too �distressed� (apologies for anthropomorphising) to be in contact with hydrophobic surface. If you�re interested in this sort of thing then take a look at our article on alkane/water partition coefficients to see how accepting a hydrogen bond (from methanol which we used to model octanol) is likely to affect the ability of carbonyl oxygen to accept a second hydrogen bond.

To be fair, the authors do recognise that accepting a hydrogen bond might affect the probability of a sulfonyl oxygen atom making contact with hydrophobic surface. However, this hydrogen bond donor doesn�t need to come from the protein or a water molecule that the crystallographers can see. How many of the sulfonyl oxygen atoms which lack �visible� hydrogen bonds are sufficiently solvent-exposed to accept hydrogen bonds from �invisible� solvent water molecules? I�ll leave it to you the reader to think about whether analysis of the CSD (as opposed the PDB) has any relevance to hydrophobic interactions.

I�m now going to wrap up with what will be seen by some to be nitpicking although that is not my intention. This is what the authors have to say about QM calculations and hydrogen bonding:

�Where experimental data are not available, acceptor strengths can be obtained from quantum chemical calculations.�

My first criticism of this statement (which is getting very close to nitpicking although as The Blogger I�m allowed to do that) is that quantum chemical calculations can be used to predict donor strengths as well. Like they might say in Buenos Aires, it takes two to tango. My second criticism is that rather than talking about generic �quantum chemical calculations� the authors could also have mentioned that electrostatic potential is a useful predictor of both acceptor and donor strength. I have to declare an interest here as author of reference 98d but I do believe that the effectiveness of electrostatic potential as a predictor of donor and acceptor strength is more important than whether it was calculated quantum mechanically or classically. It tells us something about the nature of the hydrogen bond.

That brings us to the end of my review. The article is definitely a good read and a valuable contribution to the field. To put it bluntly, you need to know this stuff if you want to succeed in FBDD. I�ve flagged up the issue of sulfonyl oxygen hydrogen bonding to hopefully make you think a bit and maybe even generate some discussion. Feel free to make comments of your own.

Literature cited

Bissantz, Kuhn & Stahl, A Medicinal Chemist�s Guide to Molecular Interactions. J. Med. Chem. 2010, 53, 5061-5084 DOI

Davis & Teague, Hydrogen Bonding, Hydrophobic Interactions, and Failure of the Rigid Receptor Hypothesis Angew. Chem. 1999, 38 736-749 DOI

Bethel et al, Design of selective Cathepsin inhibitors. Bioorg. Med. Chem. Lett. 2009, 19, 4622-4625 DOI

Toulmin, Wood & Kenny, Toward Prediction of Alkane/Water Partition Coefficients. J. Med. Chem. 2008 51, 3720-3730 DOI

Kenny, Hydrogen bonding, electrostatic potential and molecular design. J. Chem. Inf. Model. 2009, 49, 1234-1244 DOI

A short update

I�m now back in the UK and have been catching up with a few folk. It certainly was a great trip and I�ve set up a travel blog called The Great Escape (better late than never) to share some photos. So far the first month, which included a couple of weeks in Paraguay, has been written up.

You may remember the post on SPR from a while back. It got turned into a letter to the Journal of Molecular Recognition which (to my great amusement) is treated as a publication by Google Scholar. It turned out the review that inspired the post had a few folk spitting feathers and I got the chance to join the fun. I�ve never figured out why people try to eat feathers.

FBDD in Academia 2

Previously I noted that FBDD provides a means for academic groups (and start ups) to negate the advantage that Big Pharma�s massive screening collections give them. Fragment screening and structural characterisation of fragment binding broadens the scope of a structural biology group�s activities. I do believe that a package of fragment binding modes and affinities is something in which a pharmaceutical company would be interested. But what if an academic group wants to move the fragment hits (which I refuse to call �frits� because I don�t work there any more) further along the optimisation trajectory? I�ll outline some of the issues that need to be addressed and this post is intended to stimulate discussion rather than being a last word on academic FBDD. Please feel free to comment if you�d like to challenge anything or flag up anything that�s been overlooked or oversimplified.

The post-screening phase of lead generation will generally require chemical synthesis. Academic synthetic chemists typically focus on synthesising complex natural products or developing new synthetic methodology so it can be difficult to interest them in the more mundane business of lead generation. To be fair the synthetic chemistry required for lead generation is unlikely to be of sufficient novelty or complexity to earn a graduate student the PhD in synthesis which will be his or her primary objective. There are medicinal chemists in academia but in some cases these happen to be synthetic chemists who think that medicinal chemistry is simply a branch of synthetic chemistry. Also synthetic chemists in academia tend not to be interested in molecular design. The net result is that it can be difficult for a protein structure and fragment screening group to find academic collaborators to take the project into the post-screening phase even when there may be synthetic chemists in the same institution.

As you move from screening to post-screening phases of lead generation the work becomes more multi-disciplinary. Unfortunately �multi-disciplinary� isn�t something that usually gets done well in academic institutions although the problems are often less to do with skills than with organisation (and occasionally egos). In addition to the molecular design and synthesis, it will become necessary to run assays to demonstrate that affinity translates into inhibition of the target enzyme (it�s likely to be an enzyme if you�re doing FBDD). For some targets (e.g. antibacterial or kinases) it�ll be necessary to demonstrate some cellular activity. As you approach micromolar potency you may want to check that compounds in your lead series have sufficient aqueous solubility and don�t have particular affinity for anti-targets such as hERG and CYPs. This is much less of an issue if the main objective is publication but is something to be considered if you�re hoping to flog the results of the work to a pharmaceutical company.

That last comment gets me onto a tough issue for an academic lead generation group. How might you persuade a pharmaceutical company to buy your lead series? The main problem is what you�re trying to sell is information and you�ll need to show that you�ve got something good without giving it away. Life is easier if you own the relevant intellectual property but the synthetic chemistry that needs to get done to secure patent cover is not always going to get people PhDs in synthesis. The other point to remember is that Pharma people may not be willing to look at what you�ve got under a confidentiality agreement because of potential for compromising their own IP position. This can become a serious issue if you�re trying to stake a claim for future activity against related targets (e.g. all tyrosine kinases) or to negotiate exclusivity by preventing a company from using leads from competing programs.

So far this post has focussed on the difficulties (which in Pharma-speak would re-branded as personal development opportunities by the happy-smiley folk who inhabit the HR ether) of doing post-screening fragment-based work in an academic environment. If the primary objective is publication then you can write up at any point that is convenient which means that even a small amount of synthesis can have impact. In contrast, commercial lead discovery organisations need to create a secure IP position before they can publish. When publishing affinities and structures of protein-ligand complexes it�s always worth looking out for results that have relevance that goes beyond the specific project. Examples of synthetic elaboration of a fragment leading to a change in its binding mode are particular relevant and the prototypical (low molecular complexity) nature of fragments means that differences in affinity are more easily interpreted.

Getting pharmaceutical companies interested in the output of an academic fragment project is not trivial. A lot depends on the value of the target and the quality of the leads that have been generated. However, getting to leads requires organisation and realising the value of them requires commercial awareness. Organisation is about persuading people that they�re better off working together and can take time to put in place. Commercial awareness is more difficult to acquire and it�s probably best to try to keep things as simple as possible when starting out. Although this may all seem a bit daunting it�s worth remembering that synthesised compounds will typically be novel and that synthesis can be directed away from known ligands. Also one should not forget the �supporting data� of crystal structures and measured affinities.

This is a good point to wrap up. I believe that FBDD provides an excellent framework in which to both train researchers and do high quality science. FBDD also extends the range of options available to academic researchers for collaborating with industrial partners. That's where I'm going to leave it so feel free to comment if anything that I've said (or not said) has annoyed you.

FBDD in Academia 1

I�ve now gone back to being a tourist and will be in Australia until the end of the month before heading north to Singapore and Malaysia for most of June. Feel free to get in touch if you�re based in either of those countries and would like to discuss fragment stuff or Drug Discovery in general.

Some time ago, I promised to post on FBDD in academia and really can�t keep putting this off. I�ve realised that it�s not going to be possible to squeeze everything into a single post so there should be at least one more post after this one. You should be warned that my academic career ended some years before people started to talk about FBDD so if I appear to be out of touch, it may well be because I am out of touch. Hopefully some of what I�m going to say may be of interest to some of you and please remember that this blog does allow its readers to comment.

I�ll start by making two points, both of which will be obvious to many of you. First fragment based approaches provide a means for drug discovery researchers (both in academia and start ups) to counter the advantages that Big Pharma derives from having massive screening libraries and automated compound handling. Secondly measurement of weak binding and determination of binding mode of weakly bound complexes remains a frontier area in physical biochemistry and biophysics. Remember that the power of a binding assay is defined by the weakness of the binding that can be measured reliably.

An academic group with strengths in protein structure determination and biophysical measurement of binding is well placed to contribute. I see the output of protein structural studies moving away from only determining the structure of a protein to providing a more integrated view of the protein�s �interaction potential�. One point worth making in this context is that measured thermodynamic parameters for fragment binding are particularly useful for developing and validating theoretical models because there are fewer protein-ligand contacts and it is easier to quantify conformational strain. Fragment based approaches also provide a means to validate and explore bioisosteric relationships without the need for a lot of synthesis and I�ve created a graphic showing how this might work.



Assembling and maintaining a usable screening library is likely to be a challenge or at very least an issue for most academic groups. However, a group that has established expertise in fragment screening does have some advantages in negotiating with suppliers of compounds who may value experimental characterisation of how well their compounds have behave under assay conditions. Vendors of specialist fragment libraries really should value this type of feedback and if they don�t they shouldn�t be in the business of marketing fragment libraries. I sometimes wonder if synthesis of fragments might form the basis for final year undergraduate synthesis projects which could be quite self-contained and include a molecular design component. In passing I�ll pose the question to readers from academia as to whether they think they�ve got molecular design adequately covered in courses at their universities although I�ll have to leave this topic for another post.

As we all know there is more to FBDD than fragment screening. Once you�ve found fragments that bind, tested analogues of these and determined crystal structures, you�ll need to do some synthesis. For a group whose main expertise is characterising binding and protein structure determination this may a good point to bail out and prepare the results for publication. A group with some access to synthesis may wish to take the project a bit further and publish once they�ve observed some SAR. One of the attractions of FBDD for academic researchers is that there are a number of points at which they can choose to write up the project for publication. It is also worth pointing out that FBDD provides an excellent framework to gain understanding of molecular properties and interactions between molecules. This understanding is essential if you�re planning to do molecular design the basis of which is manipulation of these properties with predictable results.

What if academic researchers want to take things further and generate lead series that will be of interest to Pharma? Synthesis will be necessary and life will get more complicated. I�ll pick this up in the next post (Kakadu salties permitting) since there�s quite a bit to say and I�m actually still thinking about this.

OSDD CREATES HISTORY


The first ever open source conference in which people from around india participated and were given laptops

Melbourne, FBDD & Facebook

I now have less than a month left in Melbourne and since we�ve just switched over to winter time perhaps the hint should be taken. It has certainly been fun and the project is nicely under control although past experience suggests that it�s usually not a good idea to say that. I�ve not lived in a city since the mid-80s when I was a post-doc in Minneapolis and have really enjoyed the ease of getting around and ready access some of the wide range of music that Melbourne offers. I enjoyed an excellent performance by the ACO Soloists at Hamer Hall and am hoping to return there for a strong dose of Bach in a week�s time. University College, where I�m currently staying, is running a concert series and the second of these promises to be as enjoyable as the first. One of the music tutors at UC plays flute in the VYSO and I got to see them in action last weekend with their truly awesome guest soloist Kana Ohashi. This was also an excellent opportunity to watch the violinists since I was in the third row. The soloist was truly kinetic (difficult to be otherwise with the Tchaikovsky) and the first violin nearest to me appeared to have been given a special 'first violin bob' by her hairdresser. Maybe they will patent it.

On my first trip to the Paris Cat Jazz Club, I found it closed due to flooding (it was the day of The Hailstorm but at least there were back-to-back episodes of Hogan�s Heroes on TV). On returning the following week I was lucky enough to catch Monique diMattina who is an extremely warm, engaging and talented performer. By the way she also writes and composes and, as luck would have it, will be returning there the week AFTER I leave Melbourne. While wandering round town one Saturday afternoon, I caught The Wishing Well on Bourke Street and their next local gig is also the week after I leave. Good reasons to come back, I guess.

Previously, I pointed you towards some LinkedIn groups that are particularly relevant to FBDD. There are also groups on facebook that you might want to take a look at. It�s a bit more difficult to keep discussions going using the facebook groups because you don�t get alerted by email in the same way that you do with LinkedIn. However there are a lot of folk on facebook (especially in universities) and I believe it can play a useful part in extending the FBDD web. Here is a selection of facebook groups that you may find useful:

Fragment Based Drug Discovery (This is the group that is linked to this blog. I do check it frequently and usually respond to queries.)

Crystallography Rocks (Once you�ve got fragments to bind, you�ll want to see how they bind.)

NMR (There are a number of elegant NMR techniques for detection of ligand binding and you�ll find plenty of expertise in this group.)

Chemoinformatics (Particularly relevant to screening library design)

Dan gave my round the world trip a very flattering mention at Practical Fragments which did remind me that I really need to do a post on FBDD in academia since Teddy (who used facebook to tell me where Rapamycin comes from) has also discussed this. As I�m a real sucker for peer pressure, I do promise to make sure that my next blog post focuses on this topic. The FBDD facebook group led to me giving a lecture (I normally call these harangues) in Santiago and through it I�ve also made a couple of contacts in Singapore where I�ll probably do a couple of talks. Being in a facebook group also got me a chance to look round the Australian Synchrotron during maintenance week, when you can get a better look at all the cool stuff. I�ll finish with some pics from that visit.

FBDD and Networking

Reading an account of the session at the ACS on application of computational methods to FBDD, reminded me that it would be a good time to raise awareness of networking groups in this area. Both this blog and Practical Fragments allow readers to comment on posts although this tends not to happen with the frequency that it does at In the Pipeline, probably reflecting the huge readership, frequent updating and diverse content of what I consider to be the best drug discovery blog by a long way.

People interested in FBDD may already belong to a number of relevant LinkedIn groups. The groups offer some advantages over blogs for getting discussions going in that anyone can start a discussion and group members get alerted by email whenever somebody makes a new comment. I�ll list some of these below in case there are some that you�ve not yet heard about.

Fragment Based Drug Discovery (This group is linked by both FBDD blogs)

Label Free Assay Technology Group (It is the assay that makes FBDD possible. The weaker the binding that you can measure reliably, the more powerful your assay)

Structural Biology (X-ray Crystallography, NMR Spectroscopy, Electron Microscopy) (Generally you�re going to need crystal structures to take fragment hits forward)

Job opportunities in Computational Chemistry and Biology, Xray Crystallography, Fragment Based DD

Recently, I submitted the same item for discussion at a number of LinkedIn groups. I invited group members to share their views on the most appropriate technologies for detecting fragment binding. I learned about some new ways to configure SPR experiments and the use of Tm-shift assays. Most of the discussion was in the Structural Biology group (see discussion) although there was helpful input from the relatively new Label Free Assay Technology Group (see discussion) so thank you to all the participants. It was also great to see a couple of familiar faces from my days in Big Pharma, including a co-author from an article that a number of us wrote back in 2007

Surface Plasmon Resonance

General Reviews

Rich & Myszka, Grading the commercial optical biosensor literature � Class of 2008: �The Mighty Binders� J. Mol. Recognit. 2010, 23, 1-64 Link | Review

Application to Fragment Screening

Perspicace et al, Fragment-Based Screening Using Surface Plasmon Resonance Technology, J. Biomol. Screen. 2009, 14, 337-349 DOI | Review

Binding Pathologies

Giannetti et al, Surface Plasmon Resonance Based Assay for the Detection and Characterization of Promiscuous Inhibitors, J. Med. Chem. 2008, 51, 574-580 DOI | Review