CALAA-01 Results Indicate RNAi Therapeutics to Revolutionize Solid Cancer Treatment

In yet another important proof point that RNAi Therapeutics is slowly but surely morphing into a therapeutic reality, Calando just reported in the highly prestigious journal Nature an interim analysis of their phase I clinical study of CALAA-01 demonstrating an RNAi mechanism of action in solid cancer patients. The results are the most direct demonstration of RNAi in Man thus far, and, at least equally significant, show that the Enhanced Permeability and Retention (EPR) effect is indeed about to turn into the Achilles' Heel of solid cancers in the era of nanoparticle siRNA delivery.

Solid cancers are considered by many the most important near-term value driver for RNAi Therapeutics. The fact that Calando’s RONDEL nanoparticle delivery technology has now been shown to accumulate in solid cancer tissues is not only an important de-risking event for CALAA-01 itself, but it immediately suggest that RONDEL, and also quite likely other nanoparticle-siRNA delivery technologies, can facilitate the targeting of essentially any gene in solid cancers. It is this flexibility of the platform that makes RNAi Therapeutics ideally suited to satisfy the demands of a genetically such heterogeneous disease group as cancer.

Nanoparticle-siRNA delivery such as RONDEL is currently the most promising approach for treating solid cancers with RNAi Therapeutics. All of these rely on the particles being able to circulate sufficiently long in the blood stream and being small enough so that they can passively accumulate through EPR in the solid tumors as these lack an effective way of draining them through lymphatics. While the absolute amount of RONDEL particles that accumulated in the tumors remains to be determined, the Nature study clearly showed that a decent amount of them does accumulate there at ~0.6mg/kg.

While particle stability and ability to accumulate through EPR is a critical first step where we should also expect to see important differentiation between the technologies, once there, the nanoparticles then have to be taken up by the cancer cells and moreover mediate the cytoplasmic release of the RNAi trigger into the cytoplasm. Once again, RONDEL meets this challenge as it was possible to detect cleaved target mRNA characteristic of an RNAi mechanism of action. Although the so called RACE assay used for this analysis is not quantitative, in my experience, you have to get sufficiently robust RNAi to cleanly detect such products. Consistent with this notion, well over 50% target knockdown was detected for CALAA-01, although the limited number of samples (3) and the reliance on often only historical negative controls leaves open the question of the exact knockdown potency of CALAA-01.

It is in fact the pioneering work with RONDEL-siRNA delivery from which this pharmacokinetic model has been derived and that is now informing quite a bit our thinking of how to design siRNA-nanoparticles for solid cancer applications. For example, it was work on CALAA-01 that showed that the utility of the (transferrin) targeting ligand is in enhancing the cellular uptake of the nanoparticles, not so much in concentrating them in the tumors in the first place, and should be influential in the selection of targeting ligands being contemplated now for all types of delivery technologies, including SNALPs.


Summary of CALAA-01 results


CALAA-01 was also the first serious RNAi Therapeutics candidate for solid cancers that had entered the clinic in 2008. For a more thorough molecular background on this program, please refer to a previous entry (here) that was part of a 3-part collaborative series with Tobias Wolfram on the first RNAi Therapeutics solid cancer candidates. Briefly, CALAA-01 is a self-assembled nanoparticle system based on a cationic co-polymer containing cyclodextrin complexed with siRNA targeting RRM2, a gene involved in DNA replication, and that like most of the advanced nanoparticle-siRNA systems is stabilized through pegylation (some of these carry the transferrin targeting ligand). While Tobias and I had cautioned about a relative lack of preclinical in vivo characterization of CALAA-01, particularly not having directly demonstrated that CALAA-01 can knock down genes via RNAi in solid tumors following intravenous administration, it seems that the gamble of skipping some of the pre-clinical proof-of-concept has paid off for the investigators and Arrowhead Research (of which Calando is a subsidiary).

It was also a clever move that, amongst other tumor types, a number of melanoma patients were enrolled as such tumors can be easily accessed through biopsy and thus represent good material for molecular analysis of a clinical candidate. 3 of these volunteered to be biopsied. Despite the limited sample size, a number of conclusions can be drawn from the data: 1) RONDEL-siRNAs accumulated in the tumor tissues in a dose-dependent manner; 2) consistent with this, in the patient that received the highest dose, an RNAi mechanism of action was unambiguously confirmed by 5’ RACE; 3) this effect was long-lasting (>4 weeks) and accompanied by mRNA knockdown and disease stabilization; 4) RONDEL was safe and well tolerated in a study that involved repeat administrations and dose-limited toxicity has yet to be reached (right now at ~0.6mg/kg with dose escalation ongoing).

Clearly, these are highly encouraging results and call for the continuation of the phase I studies and further investigation of the modular RONDEL delivery platform. Importantly for shareholders, it should put Arrowhead in a better position to fulfill their stated goal of monetizing RONDEL either through licensing or outright sale. Part of the value would derive from the pioneering position of the RONDEL technology and from representing a complementary approach to lipid-based delivery which is currently dominating RNAi Therapeutics development programs.

Implications beyond CALAA-01 and RONDEL

Taking a step back, the CALAA-01 results add to the increasing amount of clinical data that first of all lay to rest concerns that there is a prohibitive fundamental (acute) toxicity issue for RNAi Therapeutics per se, and secondly provide evidence that therapeutic reality is within the reach of current systemic RNAi delivery technologies. They come on the heels of SNALP-ApoB results that showed preliminary signs of RNAi efficacy at a similar dose level while ongoing improvements in SNALP efficacy continue to widen its therapeutic index, and continuing dose escalations for both Alnylam’s VSP-02 and Silence Therapeutics’ Atu-027 solid cancer programs which can be interpreted as confirming the tolerability of some of the important delivery technologies in the space (SNALP and Atuplex).

With such progress, urgency to invest in RNAi Therapeutics, particularly in enabling delivery technologies should return. As the transition into the clinic is fraught with uncertainties for new chemistries (safety, efficacy, and manufacturing scale-up), it is particularly the less than a handful of delivery technologies that have made it there for which a land-grab might erupt as is also indicated by the fact that Tekmira now counts 8 RNAi-related pharmaceutical collaborators (both early and advanced stage), mdRNA at least 3 such early-stage collaborators, and a rumor-driven rise in the shares of RXi Pharmaceuticals possibly due to their mysterious claims on ‘self-delivering’ siRNAs. With the publication of the results that are about to garner widespread attention, Arrowhead Research/Calando should, of course, also get a few enquiries for RONDEL as it resumes its search for partners. Considering the generally weak share prices of RNAi Therapeutics companies, all the ingredients seem to be in place to see related deal activities, with a significant portion of the value being driven by the solid cancer opportunity for RNAi Therapeutics.

Update May 29, 2010: More details about the study, including the safety profile was published recently in an abstract for the upcoming 2010 ASCO cancer conference:

Systemic delivery of siRNA via targeted nanoparticles in patients with cancer: Results from a first-in-class phase I clinical trial

Author(s): A. Ribas and colleagues

Background: Systemically delivered small interfering RNA (siRNA) would allow targeting oncogenic molecules beyond current approaches. We report on the first siRNA trial with a targeted nanoparticle delivery system. Methods: Open-label, dose- escalation trial in pts with solid refractory cancers receiving 4 i.v. infusions (30 min) on d 1, 3, 8, and 10 of 21-d cycles. CALAA-01 nanoparticles consisted of a cyclodextrin-based polymer, transferrin protein (hTf) targeting ligand, polyethylene glycol (PEG) for stability, and siRNA against ribonucleotide reductase M2 (RRM2). The 70 nm particles were designed to minimize renal clearance and allow tumor vasculature permeation with binding to tumor hTf receptors (TfR). Primary endpoints (safety, MTD determination) were based on the first cycle. Results: 15 pts accrued to 5 dose levels (3, 9, 18, 24, 30 mg/m²), median age 62 (range 53-85). Most common histologies: GI (4), melanoma (3). Dose escalation progressed with no DLTs. Most common treatment-related AEs: fatigue (47%), fever/chills (33%), allergic (33%), constipation (33%), nausea/vomiting (20%), all g1-2. 1 pt had g3 anemia and 2 pts had g2 thrombocytopenia, all shortly after CALAA-01 infusions with rapid recovery. 1 pt had possibly-related sinus bradycardia (g2). No objective tumor responses were seen; 1 pt at highest dose had stable metastatic melanoma for 4 mo, a change from prior course. Biopsies in 3 pts (melanoma) showed particles in tumors. At 30 mg/m² RRM2 knockdown (mRNA and protein) was seen with confirmation of mechanism by specific cleavage sequence (RACE-PCR). TfR was not downregulated in cancer cells.Conclusions: Systemic delivery of siRNA via targeted nanoparticles is safe and can induce specific, siRNA-mediated gene silencing. This approach could be expanded to any currently undruggable cancer therapy target.

For Silence Therapeutics’ Atu-027 Cancer RNAi Therapeutic, the Proof is in the Pudding, not the Theory

This is the second part of the three-part series in which Tobias Wolfram and I are looking at the scientific merits of the first three notable RNAi Therapeutics candidates that have entered the clinic for solid cancer indications. Solid cancer is arguably the one therapeutic area where RNAi Therapeutics currently have highest value because of a) the ability to target many of the important, but otherwise undruggable cancer-related genes; b) systemic delivery technologies that ought to be able to reach solid cancer tissues; and c) a patient population that is essentially indifferent to the exact mode of administration (e.g. oral vs intravenous infusion).

In Part I of our series we looked at Calando’s CALAA-01. We came out slightly disappointed with that program because it seemed stuck in the quite early, non-biological/physico-chemical characterization stages and did not provide too much bona fide evidence of anti-cancer activity in pre-clinical animal models. Neither was Calando’s RONDEL delivery technology convincingly shown to facilitate in vivo gene silencing in the first place. We are therefore relieved, and I personally also somewhat pleasantly surprised at the same time, to be able to report a considerably better validated cancer RNAi Therapeutics candidate in Silence Therapeutics’ Atu-027. While the precise theory behind Atu-027 remains somewhat uncertain, we are encouraged by the scientific rigor with which the target had been validated as a component of the well known PI3-kinase pathway and the careful demonstration that lipoplex-siRNA delivery can be used for knocking down genes in the vascular endothelia from mice to monkeys.

The Target: PKN3, a Downstream Component of the Cancer-related PI3K pathway

The precursor of Silence Therapeutics, Atugen AG, was a child of the millennium genomics era involved in gene discovery and target validation. The technological workhorses underlying this work were a combination of gene knockdown approaches such as ribozymes, antisense (‘GeneBlocs’), expressed RNAi that were combined with RNA expression analysis and proteomics.

For some reason that may be related to the prior training of Atugen’s scientific staff, Atugen specialized in studying the genetic components of the PI3-kinase pathway. This pathway has attracted wide attention because it plays important roles in various biological processes and has consequently been implicated in numerous diseases, particularly in cancer where a number of genetic abnormalities that affect a range of phenotypes ranging from cell growth to metastasis have been linked to it. While clearly an attractive target for cancer therapy, and there has been some success in drug development in that regard, its many roles in molecular biology means that targeting the pathway upstream, for example at the PI3 kinase level, is likely to be associated with a poor therapeutic index. Atugen therefore reasoned that discovering targets downstream of PI3K could result in equally potent, but also more specific therapeutics for cancer.

Applying their genomic tool kit that placed a high priority on specificity controls, they came up with a handful of candidate target genes that they then nicely validated in subsequent in vitro and in vivo tumor-related assays. From this, PKN3 eventually emerged as a gene whose expression and phosphorylation was not only PI3 kinase-dependent, but when knocked down also abolished the ability of PI3 kinase-activated cancer cells to grow under culture conditions thought to reflect the natural tumor environment. When such cancer cells were modified with a DNA-directed RNAi construct against PKN3 and then introduced into mice in an orthotopic prostate tumor model, the primary tumor formation was, somewhat unexpectedly, not much affected whereas metastasis formation was significantly inhibited. Thus, the authors speculated that it is PI3 kinase-dependent functions like cell motility and/or cell-cell/cell-matrix contacts that are disturbed following PKN3 knockdown, and that the substrate-dependent growth defect observed in vitro may reflect deficiencies in tissue invasion that are shared with the facilitate metastasis in vivo. Consistent with a role for PKN3 in real-life cancers, tissue immunohistochemistry from prostate cancer patients revealed PKN3 to be upregulated in the cancer, but not surrounding normal tissue.

The theory of why PKN3 RNAi might have utility as a cancer treatment had to be modified when it was found that the delivery system did not target the cancer cells in which the initial target validation experiments had been carried out, but instead endothelial cells of the blood vasculature. The late change in the theoretical framework for Atu-027 that occurred in the otherwise very impressive 2009 landmark Atu-027 mouse-to-monkey validation study (Aleku et al., 2009), is probably the major weakness in Atu-027’s rationale. The question arises why take such a target risk at all when there are many other well-validated, but undruggable cancer targets already out there to choose from? Is PKN3 a suitable cancer target after all, and, even if it were, are they knocking it down in the right cell types? It is here that the relative lack of knowledge on the biology of PKN3 (all the functional studies on this gene were conducted by Silence Therapeutic) turns out to be a disadvantage and may complicate things like the development of biomarker tests to monitor early clinical efficacy.

On the other hand, the fact that PKN3 emerged after having considered a range of targets, suggests that based on practical experience Silence had highest confidence in the effectiveness of PKN3 as a cancer target. This argument is supported by what Tobias and I felt to be very well controlled target validation experiments, including the use of multiple positive and negative control oligonucleotides to confirm that the observed phenotypes were on-target. In that regard, I would score the relevant dataset generated by Silence in the top 5% of the RNAi literature. Maybe not surprising because of Silence's roots in gene discovery.

Being the only one working on a gene, however, has a few benefits, too. As such, Silence has made inroads in obtaining broad patent protection for the use of PKN3 as a target in PI3 kinase-related diseases. Given the large interest in the PI3 kinase pathway, this could be a valuable asset to the company and somewhat further justifies what otherwise would look like a disproportionate investment for a platform company into a single pathway!

The RNAi Trigger: A 23 base-pair Atu-siRNA

Although Atugen’s background was more in ribozymes and antisense, it decided to make drugs based on synthetic siRNAs after its transformation into a drug development company. This was around the time that RNAi had just been discovered in humans by Tuschl. In the case of Atu-027 this meant that while antisense, expressed RNAi, and small molecules acting on the PI3 kinase pathway were used for target validation, the active ingredient would be a 23 base-pair blunt-end siRNA in which alternating 2’-O-methyl nucleotides would face unmodified residues, a pattern that also coincides with the eventual scope of the issued Atu-siRNA patents in the US and Europe.

Since siRNAs may have anti-tumor activity independent of RNAi, the result of non-specific innate immunostimulation by some siRNAs, it is notable that such extensive 2’-O-methylation was shown only later in the field to result in siRNAs with no or very little innate immunostimulatory potential. This not only lends credibility to their pre-clinical efficacy data, the choice of 2’-O-methylation at such an early stage can therefore be more generally regarded as a very lucky one considering that the scope of the Atu-siRNA patents would eventually only encompass 2’-O-methylations. Speaking of RNAi trigger IP, a strong Tuschl II would actually benefit the blunt-end Atu-027, while a strong 21-23 base-pair/nucleotide covering Tuschl I could cause problems.

Lipoplex-siRNA Delivery to Endothelial Cells of Blood Vasculature

The delivery system for Atu-027 was developed in parallel to identifying the target siRNA. It is a pegylated lipoplex-siRNA system where the siRNA sits on the outside of positively charged, peylated liposomes ('Atuplex'), instead of the more commonly used SNALP-like liposomes where the siRNA is encapsulated inside. Because of this fundamental difference, the two systems vary considerably in their biological properties. Whereas SNALPs deliver their cargo mainly to the liver, but can be further stabilized to reach tissues beyond that, lipoplex-siRNAs have a preference for the endothelia lining the inside of blood vessels pretty much throughout the body.

In careful labeling studies, Silence Therapeutics scientists were able to show convincingly that they can achieve bona fide RNAi knockdown in the 50-70% range when administering around 1mg/kg- from mice to monkeys. Moreover, the invariable pharmacokinetics following repeat administration support that there is little or no cytokine activation. It is curious that most of their in vivo studies involve repeat administration. This is in contrast to what for example Tekmira and Alnylam are practicing which carefully study both single and repeat administration both of which together should inform much better on eventual dosing schedules, causes for toxicities etc. Similarly, only one cationic lipid, AtuFECT01, is used throughout the years, and a relatively simple opportunity to improve the therapeutic index has probably been missed. There is also a lack of a detailed description of the precise formulation process and early-stage physico-chemical characterization, although part of this may be for competitive reasons.

Apparently limiting themselves to delivery systems developed in-house, the performance of the lipoplex-siRNA technology means that in order for Silence to pursue their long-standing interest in cancer, they have to address the disease where it interfaces with blood vessel endothelia. Importantly, the concept of starving cancer cells by inhibiting neoangiogenesis is well accepted and one only has to look at the commercial success of Roche/Genentech’s anti-VEGF monoclonal antibody Avastin. The more pressing task therefore is to find the right target genes.

Pre-clinical safety and efficacy

The lipoplex-siRNA formulations were generally well tolerated at dosages predicted to be therapeutic. No significant differences in body weights were observed with various siRNAs. Immune responses were also not detected. This is particularly important in studies like these as it further suggests that the observed anticancer activity was not due to an off-target immune-related mechanism. Further supporting such a lack of immune responses was, as noted above, the fact that the pharmacokinetics did not change with repeat administration. Immune reactions would have likely facilitated antibody formation against the PEG component and caused rapid clearance of the lipoplex-siRNA upon repeat administration. I originally had been skeptical of Silence’s claims that there were no immune issues since they never seemed to have paid much attention to this issue, but they may have just gotten lucky in their early choice of extensive 2’-O-methylation as part of the Atu-siRNA architecture.

There were, however, some toxicities noted that mostly involved the liver and included 2-3 fold minor elevations in liver enzymes with an PTEN siRNA (this could be an siRNA-specific response different from Atu-027) and an apparent increase in DNA replication in livers of mice treated with control lipids alone which may be indicative of liver cell damage (Santel et al.,2006). Liver safety and immune reactions may therefore be the two expected dose-limiting adverse events for Atu-027.

In terms of efficacy, Atu-027 had ED50s in mice and monkeys of 1 and 0.3mg/kg using liver and lung endothelia as surrogate tissues. Since the samples include non-endothelial contaminants that could express some PKN3, this may actually somewhat underestimate the true extent of the knockdown in the target endothelial cells. This knockdown efficacy translated into a 60% reduction of the primary tumor in an orthotopic mouse prostate cancer model, and an even greater 80% reduction in the lymph node metastases. It should be noted that while the effect on the primary tumor is to be welcome, the results somewhat differ from those in the initial characterization of PKN3 as a target 5 years earlier where an effect was only observed for the metastases (Leenders et al., 2004). The anti-tumor response was confirmed in a range of other tumor models. Taken together, the safety and efficacy profile of Atu-027 are in support of initiating phase I studies.

Target and Delivery: A match made in heaven, or marriage of necessity?

As alluded to in the target validation segment, we are somewhat concerned about target risk since, as a result of the emerging properties of the lipoplex-siRNA system and in vivo PKN3 knockdown phenotypes, the rationale for PKN3 as an anticancer target has undergone multiple transformations, first validated as playing an important role in the cancer cells itself, but then thought to act directly on blood vessel and eventually lymph vessel formation.

Cynics could say that once they got stuck with the lipoplex-siRNA system, they would have done anything to force the model to fit their data. Realizing this dilemma, they did, however, undertake bridging studies and were for example able to show that PKN3 knockdown in isolated (non-cancer) vascular and lymphatic endothelial cells disturb well known endothelial phenotypes such as ability to form tubes in tissue culture.

Despite these twists and turns, from a 30,000 foot level, PKN3, due to its well-validated role in the PI3 kinase pathway, does look like a credible anticancer target in endothelial cells. Are there potentially targets that are better suited for the lipoplex siRNA system? Most likely yes, as it ought to be a general rule in RNAi Therapeutics that ‘target follows delivery’. Thus armed with knowledge about the ‘Atuplex’ delivery system, the next target-delivery combination by Silence has a better chance of looking like a match made in heaven.

Conclusion

In the end, we came to the conclusion that, theoretical concerns notwithstanding, by targeting a skillfully validated PI3K pathway gene with a delivery system that facilitated bona fide gene knockdown from mice to monkeys and the clear anticancer phenotypes observed in a number of rodent cancer models, Atu-027 is a promising clinical candidate for which the pre-clinical proof was simply in the pudding. Given the pioneering and innovative nature of this particular approach, it should also be able to draw the attention of the likes of Novartis and Pfizer that should be able and willing to take such risks as they move into RNAi Therapeutics.

Atu-027 may also be viewed as the start of a franchise that focuses on attacking cancer from its endothelial side with all the unique advantages of RNAi, for example the almost unlimited choice of targets. The merger with Intradigm, which also has developed expertise in neoangiogenesis and cancer, should add depth to this by, most importantly, expanding the pool of potential targets and pathways so that potentially less risky target-delivery matches can be found. There should also be ample scope for improving the Atuplex delivery system as it is questionable whether the first cationic lipid that they published and held onto, Atu-FECT01, is really the non plus ultra of lipid chemistry.

Meanwhile, Atu-027 has started clinical development half a year ago with a multi-dose phase I study in patients with advanced solid tumors. First results are eagerly awaited.

Calando’s RONDEL RNAi Therapeutics Delivery Promising, but CALAA-01 Prematurely Entered into Clinic

[Part 1 of a 3-part collaborative series with Tobias Wolfram on the first notable attempts at RNAi Therapeutics for solid cancers that have entered the clinic]

When during the company's R&D day the CEO of Alnylam, John Maraganore, highlighted Calando’s cyclodextrin-based siRNA delivery technology (RONDEL) as one of the noteworthy non-SNALP systemic siRNA delivery technologies out there, it certainly piqued my interest. This is not least because any cash infusion and longer-term commitment by a partner like Alnylam would do wonders for the parent company of Calando, Arrowhead Research which is a conglomerate of early-stage, IP-focused business units and has just barely scraped by bankruptcy through a diet of cutbacks. While I always remembered the maturity of RONDEL, developed in the Mark Davis lab at Caltech, to be years behind SNALP, Tobias and I decided to to take a closer look at the development path of CALAA-01, the first clinical RONDEL delivery candidate and also investment focus of reorganized Arrowhead Research.

When Tobias first heard of the technology, it struck him as a very elegant, because simple, and modular method to formulate targeted nanoparticles. In fact, there are not many targeted nanoparticle siRNA delivery approaches where the components supposedly can be assembled by the pharmacist just before patient administration. RONDEL siRNA delivery consists of mixing together siRNA, a short cyclodextrin-containing polycation, and adamantane-coupled PEG stabilizers some of which carry a transferrin ligand, so as to create 60-80nm particles. These particles were rationally conceived to satisfy a range of pharmacologic and formulation considerations. Their suitability for solid cancer relies on the enhanced permeability and retention (EPR) effect of nanoparticles with reasonably long circulation times (here supposedly achieved by PEG stabilization), the ability of the particles to be taken up into cancer cells by transferrin receptor-mediated endocytosis and their subsequent release into the cytoplasm in a pH-dependent manner.

Unfortunately, what we soon came to realize was that while the concept is a very nice one indeed, the particles, particularly CALAA-01, remain to be better characterized both physically (shape, uniformity, storage and biological stability etc) and for their RNAi knockdown ability in vivo. For example, knockdown of RRM2, the target of CALAA-01, and subsequent tumor inhibition have not been demonstrated in a convincing in vivo system. Instead, knockdown efficiencies have largely been limited to in vitro studies. Moreover, these involved siRNAs that were selected with what today would be considered outdated methods and probably as a result were not very potent. The in vivo studies were essentially limited to pharmacological investigations, with the combination of in vitro efficacy and in vivo pharmacology forming the rationale for moving CALAA-01 into the clinic. Moreover, even when considering only the pharmacology, measures such as biodistributions and circulation times did not fit the model which may be explained by nanoparticle instability in vivo, something that really needs to be investigated further. Also, since the siRNAs were unmodified it strikes me as rather strange that no innate immune induction and only moderate adaptive immunity were reported.

What I found to be a valuable take-home message from those studies, although according to Tobias’ liking resting too much on indirect evidence, but not necessarily data obtained with the CALAA-01 clinical candidate, was that the utility of the targeting ligand appeared to be in increasing the cellular uptake of particles with little positive surface charge, less so in skewing the biodistribution towards the solid cancers per se. This could also be because the particles were cleared relatively rapidly from circulation, mostly into the kidney and bladder, which raises further questions about the purity and integrity of the particles. In this light, the mention of nanoparticle assembly by a pharmacist may also be interpreted a necessity due to storage problems of fully formulated particles. Nanoparticle assembly is notoriously sensitive to even slight changes in parameters such as temperature, speed of mixing etc so that it would be preferable for the physician to just administer the drug without the need for prior handling. It is therefore unfortunate that results from long-term storage and robustness of the formulation method were not presented. Nevertheless, an enhanced cellular uptake through the addition of a ligand could critically increase the therapeutic index of a cancer RNAi Therapeutic, and the modular nature of the RONDEL system should easily facilitate such additions.

Our assessment that CALAA-01 was probably entered into the clinic too early naturally rests on the publicly available data only. The publication dates of the relevant data, however, strongly suggest that they indeed represent the relevant data points and considering the financial situation of Calando, it would not appear that much were to be gained by holding back on positive data. This is by no means to belittle what otherwise is very rich science. Unfortunately, it is here that the tension between corporate demands for advancing a pipeline and the need to sufficiently advance the science is most evident and in the end risks harming both objectives. Optimistically, completion and evaluation of the CALAA-01 phase I trial will allow for valuable insights into the performance of RONDEL delivery in man for the benefit of any follow-up programs. Without any such strong data, it is questionable however whether the promise of RONDEL as a differentiated and flexible platform for RNAi Therapeutics delivery alone will be enough to make Arrowhead Research a good RNAi Therapeutics investment.

PS: To expand on the latter point, Tobias and I also discussed that, in general, it is easy to caution against entering RNAi Therapeutics candidates into the clinic early and dismiss such as a move of desperation. On the other hand, the case can be made that, when it comes to RNAi Therapeutics as a broadly applicable platform, clinically evaluating candidates which for example would not be expected to effect large knockdowns can be justified in that the data coming out of these studies may provide timely data invaluable for follow-up programs using very similar delivery approaches. Although investors will rightly fear the costs of a failed trial particularly for small biotech companies, such data may be valued more highly by a potential Big Pharma partner. This argument receives added weight in an environment like now where it is very difficult to raise capital from the public markets, and partnering is the primary means for small biotech to obtain capital at acceptable terms.

Combining Forces to Understand RNAi Therapeutics Inside Out

This entry is first a PR on a business development of the blog itself, but also reflects what should be the next major value-driver in RNAi Therapeutics after the liver: solid cancers.

Solid cancers are a very attractive target for RNAi Therapeutics because of its unmet needs and because of the so called Enhanced Permeability and Retention (EPR) effect of solid tumors which means that it should be possible to target them with siRNA-containing nanoparticles. Getting there, of course, is only half the story. The particles need to navigate their way through the extracellular matrix of tumors, latch onto the cancer cells, be taken up, and then finally be released into the cytoplasm. This requires a good understanding of nanoparticle-related chemistry and physiology, the biology of the extracellular matrix of cancer tissues, and cancer cell membrane biology.

Enter Tobias Wolfram. I have known Tobias since my days studying biology in Heidelberg (10 years ago now!) and have since been impressed by his enzyclopaedic knowledge of not only biology, but also history, psychology, economics and what not. He spent much of his time as a teenager with science projects and in molecular biology labs and won national prizes, at a time I did not even know that PCR existed. Since then he has become a truly multi-discliplinary scientist spanning the subjects of biology, chemistry, and material physics. Right now, he is at the Max-Planck-Institute for Metals Research in Stuttgart, the home of German engineering, where Tobias employs precisely engineered nanometer-patterned substrates for studying the interaction of cells with the extracellular matrix and their use for cell-based diagnostics. I visited him there two weeks ago to talk about working more closely together on the topic of RNAi trigger delivery.

In an experiment, we have decided to combine his expertise in getting molecules to cells with my understanding of the molecular biology of RNAi inside cells, to hopefully provide more insightful blog entries on the topic of RNAi Therapeutics delivery, with an initial emphasis on solid cancers. With Calando, Alnylam, and Silence Therapeutics having active programs in solid cancers, we will, over the next couple of weeks, start by taking a look at each of the applied technologies.