If you are claiming about a "novel" anticancer compounds, you should first go in a simple in vivo model and see whether your compound keep any "significant" activity.

You have to keep in mind that a cancer is much more than its sole cancerous components. Today, "novel" anticancer compounds that "target" "multitarget components within the tumor microenvironment" have great chances to reach actual clinical development (see the attached article). This is by far another story than killing some cancer cells in a flask.

Once you have a proof of in vivo activity, in a simple in vivo model, but representative of the pathway you aim to study, then you will have to perform various in vitro experiments to rule out "off-targets".

Thus, there is "none best way" to validate the mode of action of a "novel" anticancer compounds, at least in vitro ... It is also highly depending about the journal you hav in mind to publish your data ...

Best regards

Robert

This is possibly true, as some pharmacological agents can have biphasic activity. The biphasic activity is usually/often associated with concentration dependency and contextual application. Resveratrol is a good classical example.

Firstly are you concerned about the mode of action or the mechanism of action.

Mode of action is only the effect seen physiologically/ anatomically as a result of the mechanism through which your compound acted. From your example of article i assume you are talking about the mechanism of action which is the molecular level analysis of how the compound is doing what it is doing.

As for the mechanism of action you could do a simple analysis of the pivotal proteins like NFkB etc in the presence or absence of your compound and discover the mechanism through which the action occurs (via simple western blots for example). This would help you understand the molecules involved which in turn can be extrapolated to physiological/anatomical effects, leading us to the mode of action. 

Once you figure out a gene that is actually getting affected by the compound you could focus on doing an RT PCR with and without treatment to check whether the mechanism is transcriptional or does the change occur only in the protein levels even though there is no change in the RNA expression profile (in case he compound in some way harms the stability of the produced protein). There are many other experiments that can help you figure out what the mechanism of action of the compound is which will then lead you onto either in vitro(in cells) or in vivo (animal models) experiments to check what physiological end points are seen.

Hopefully i understood your question right and hopefully this helps

I agree with Robert Kiss. A dogma set only on mutations led us out of the way for long time. Cancer is a complex disease, where the same type of cancer has phenotypic characteristics very different even in different persons. It can rightly be regarded as a new tissue produced by an environment characterized by a lasting chronic inflammation. What usually is omitted in the scientific considerations on cancer, is a matter related to the human metabolism that is not adequately considered, either for poor familiarity of many, either because of the complexity it introduces. It is now well known that at least 40% of the proteins present in eukaryotic cells are intrinsically disordered (IDPs). The most important property of these proteins is of not possessing an organized structure over time under physiological conditions. But the most important consequence is their enormous capacity to interact with many and different molecular partners paving the way to different functional activities. This ability is even more amazing when you consider that interactomic studies have clearly shown that they have a key role in the metabolism where they occupy key points (Hub proteins) as connectors to other metabolic pathways. They are present in all cellular compartments, including nucleus.We still do not know exactly when and how a biological function is changed through a change of IDP partners, but this makes the metabolic environment indeterminate. In fact, on the large-scale, a metabolic perturbation cannot be adequately understood by means of the usual deterministic logic that is based on cause and effect. When a cell is metabolically stressed by internal or external perturbations, the metabolic response is predictable only on small scale (at enzyme level, or single signaling pathway) whereas the general metabolic vision is blurred  just because it lacks the knowledge of the new functional behavior of the IDP.  Most likely in cancer we are dealing with impairment of the IDPs, which, therefore, are the targets of choice for a drug that can effectively combat and treat cancer. Until now, the facts show that many drugs are ineffective and at best they allow some survival situation with strong side effects and a big useless waste of money.

Also I add that the only method of testing for the new compounds in cancer therapy is not.

As mentioned in the previous topic, any mutation makes the drug ineffective or specific effective (if a new drug acts on the mutant component of tumor growth).

But in this case, any mutation causes atypical cell signaling on the down

It seems to me the best way to check is the algorithm:

- In vitro

- In vivo

- Pre-clinical studies

- Phase 0

the rest is on the level of speculation (the drug can act on a cell line and not to effect the other