What is the expected result of the treatment?

The expected result is destruction of all cancer cells and the patient being cured.

How soon will the treatment take effect?

The Ambrego treatment is designed to maximize the likelihood that the drugs will be effective from the start.

A marker version of the drug makes cancer cells visible on MRI, showing whether the correct cells are being targeted. The therapeutic versions detect cells in the same way. By this stage, Ambrego Labs has already tested the active ingredient in vitro using the patient's cancer-cell sample.

After administration, the drug's effect in destroying cancer cells is detectable in blood tests within days. The effect is clearly visible in MRI within the first few weeks.

Is the treatment given in parallel with conventional chemotherapy or as monotherapy?

Ambrego treatment is intended as a complete replacement for conventional chemotherapy across most cancer types and stages. Data from tests and treatments performed to date indicate that it may be safer and more effective than conventional chemotherapy.

Even though this is the case, we recognize that conventional chemotherapy is established and proven, while our treatment is not. The safety profile of conventional chemotherapy is well known, and although it has many side effects, it will take a very long time to fully replace conventional chemotherapy.

Even with conventional chemotherapy, a cure is often uncertain or unlikely, and treatment may be intended only to prolong life. These are precisely the cases in which adding the more targeted Ambrego treatment may help turn palliative chemotherapy into an effective curative treatment.

Is there a risk of cancer recurrence? How can you be sure that you will be targeting and treating all cancerous cells?

The risk of cancer recurrence remains if the DNA mutations targeted are DNA mutations that occurred in a later generation of the cancer cells.

To reduce this risk, tissue samples must be taken from the correct locations. For example, if cancer-cell samples are taken from the original tumor, its DNA mutations will also be present in all tumors derived from it. The metastatic cancer can therefore be targeted throughout the body.

The same safeguards used to verify the treatment's effect—including assessment of targeting in vitro and in vivo—also reduce the risk of cancer recurrence.

How expensive is the treatment?

An experimental animal treatment costs between USD 30,000 and USD 100,000, depending on the work and precautions required. A human n-of-1 study, including the necessary safety measures and third-party costs, starts at several hundred thousand US dollars.

However, we take the patient's financial situation into account in our offers wherever possible. We also cover part of the costs as long as the treatment is experimental.

Why is the treatment designed to work across cancer types and in both humans and animals?

Conventional cancer drugs do not act on the actual cause of cancer, which in the vast majority of cases is DNA mutations, but on the changes in the cell caused by these mutations. These are often cancer type specific and often species specific.

The Ambrego drugs act directly on the DNA mutations with their own proteins (the tools of the cells), which are delivered to the cell as part of the drug. All animals and humans share the concept of DNA as the building instructions for the cell.

Only the proteins expressed to stop cell growth or destroy cells differ between species. Because Ambrego's technology is modular, the drug can be adapted to a different species within a day.

Why is it important to have separate drugs to stop cell growth and destroy cells?

Interrupting cell growth is the key to cancer treatment that does not stress the patient's body. If this does not happen, cancer drugs must be very aggressive and destroy more cells than have grown through the tumors in the body just to reverse the cancer's effect.

Actively destroying cells is the key to curing a patient in a controlled manner. This is particularly important for cancers in which simply interrupting cell growth has proved insufficient: in these cases, conventional chemotherapy may prolong life without curing the patient.

Why is targeting one or a few DNA mutations enough to treat all cancer cells, even if they are spread throughout the body?

Tumors arise from a single mutated cell, accumulating additional mutations as they progress. Even when tumors become metastatic, they keep the mutations from the original mutated cell.

The Ambrego treatment targets mutations that are common to all cancer cells, i.e., that have arisen from the single mutated cell. Each of these mutations separates healthy cells from cancerous cells and can be used to target the cancerous cells independently of the other mutations.

In the rare case of multiple independent cancers in the same patient, multiple DNA mutations must be targeted. Ambrego technology can do this by adding multiple guide RNAs.

Individualized cancer vaccines often target 20 different mutations (for example, BioNTech's). Why can targeting one mutation with Ambrego's treatment be more effective than targeting 20 with an individualized cancer vaccine?

Individualized cancer vaccines target multiple mutations because there is no guarantee that they will be effective in targeting a specific mutation.

Cancer vaccines do not act on the DNA mutation itself, but on the so-called "neoantigens", the proteins that are formed when the gene with the DNA mutation is expressed. However, there is no guarantee that the gene will be expressed and thus that the neoantigens will be formed, nor is there any guarantee that the neoantigens will have a surface sufficiently different from the original protein for the vaccine antibodies to distinguish the two.

Ambrego drugs, by contrast, detect DNA mutations by binding to them directly. This binding causes the active ingredients to be expressed in the cancer cells. Multiple mutations can be targeted using multiple guide RNAs, but this is not required for efficacy.

Does the Ambrego technology modify the patient’s genes in any way?

No. None of the components of the Ambrego drugs is able to modify DNA. Even the Cas9 proteins are mutated deactivated Cas9 (dCas9) proteins that are no longer able to cleave DNA.

CRISPR-Cas9 proteins are known to have off-target effects. How could these affect treatment?

The Ambrego technology incorporates the knowledge gained from years of research on these technologies at institutes around the world.

The drugs incorporate safeguards intended to limit their activity to cancer cells. These include highly specific dCas9 variants designed to bind only to DNA containing the cancer-associated point mutations. Two dCas9 proteins are also used to reduce the possibility of off-target effects further.

Together, these safeguards are intended to give the drugs a very low probability of attacking healthy cells—far lower than with chemotherapy.

What about adverse immune reactions to the drug?

The risk of adverse immune reactions depends on the drug-delivery method. With modern delivery methods, such reactions are not expected.

How does the expected efficacy of Ambrego treatment compare with that of other cancer drugs?

Because Ambrego drugs work with an active in-cell system that is delivered into the cell as part of the drug, we can extensively optimize this system to achieve exceptionally high efficacy. We will continue to do this during the experimental animal treatments and human n-of-1 studies.

Does the efficacy and safety profile of Ambrego treatment differ from patient to patient?

Because the drug is modular, we do not expect the efficacy and safety profile to vary substantially between patients. This must still be demonstrated in the first animal treatments and human n-of-1 trials.

Are there any side effects? If so, what are they like?

The Ambrego drugs are extremely targeted, and the correctness of this targeting is extensively tested for each patient with different versions of the drug before starting the actual treatment. One drug marks cancer cells with eGFP, which is visible under a microscope. This allows Ambrego Labs to test that exactly all of the cells in the patient's cancerous cells sample are targeted and none of the healthy cells are, as a quality control before sending the other drugs to the oncologist. The oncologist will then use a different version of the drug that makes cancer cells visible on MRI to check that the correct cells are being targeted via 3D MRI.

The treatment design also reduces the stress on the patient. By stopping the cell cycle of the cancer cells, cell growth is stopped and the cancer cells can be destroyed in a gentle way.

For these two reasons, we expect a substantially better side-effect profile than with chemotherapy.

Why are you pursuing n-of-1 trials instead of traditional trials that could lead to approval for general use?

The drugs are calculated and produced on a per patient basis. Still, because of the toolbox-like nature of the Ambrego drugs, we believe that they have a consistent efficacy and safety profile in patients, but we have yet to prove this.

This will be evaluated in the first human n-of-1 trials. We can then use the results to discuss the best path to standardization with the relevant authorities.

Why can't you treat cats in the EU? My cat is sick, and I need help urgently.

The EU Directive 2010/63/EU of September 22, 2010 on the protection of animals used for scientific purposes requires mice, rats, guinea pigs, golden hamsters, Chinese hamsters, Mongolian gerbils, rabbits, dogs, cats, non-human primates, frogs and zebra fish to be purpose-bred if they shall be used for experimental animal treatments.

Thus, the Ambrego treatment must first be approved for use in animals in the EU, which, while we are working towards this, does take a long time due to the individualized nature of the Ambrego drugs.