Do you want to take a chance?

Checking potential senolytics on yourself

Fight Aging!, Planning a Single Person Trial of Senolytic Drug Candidates Translation: Pattern, SENS Volunteers group, Geektimes

For links, see the original translation.

The mice in the photo are the same age, but 50% of the senescent cells were removed from the right one

This post should be considered as part of an ongoing and not yet completed process of reasoning about experiments on oneself in order to test candidates for senolitics. We are talking about substances that, as animal studies have shown, selectively destroy senescent cells to one degree or another. For some of them, a positive effect was demonstrated in various animal experiments years before the current wave of interest in senolytics, and some of the noted effects can be attributed with a certain probability to the removal of senescent cells. Some of them have been tested as potential cancer remedies, or painkillers, or for other purposes. Some have serious side effects, as do most potential chemotherapy treatments. They are designed to destroy cells, and their selectivity is far from desirable. Nevertheless, all these potential senolytics are on sale, and therefore available for conducting experiments on yourself.

Experiments have a long and vivid history in the scientific community. Many well-known researchers have obtained the first results of human trials by experimenting on themselves, and it seems to me the most ethical approach: the researcher takes risks. Distancing ourselves from the risks for a while, the main conclusion from such stories is that they are meaningless without taking measurements and publishing the experiment process and the results obtained. Attempts to guess the results or use potential drugs simply in the hope that their effects in humans will be similar to those in mice are meaningless. The same can be said about the choice of indicators based on the principle of ease of measurement. The objective goal of the experiment is to learn something new and report it, which is achievable even in the conditions of one person without a control group, provided that the desired effects manifest themselves steadily and clearly, and that the experiment was conducted correctly. In such a context, experiments on oneself can contribute to conducting methodologically more rigorous studies by groups with resources in order to more accurately determine the effects obtained, the optimal dosage, etc.

You will need the help of a doctor

First of all, you need to find a doctor who understands what you are going to do and for what purpose, and is ready to order the appropriate tests. You will need a guide to the world of the local medical community, especially for more expensive types of tests and tests. Usually, it is not difficult to get access to them, since you will order them on a paid basis.

 Find an intermediary laboratory

You will need an organization that would act as an intermediary when interacting with suppliers, since many of them do not accept orders for senolitics from individuals. In our time of restrictions on the turnover of medicines, the supply of medicines from abroad is simplified if it is carried out between laboratories. You will need to cooperate with a company that has a laboratory or that can play the role of an intermediary when contacting the laboratory to obtain the services you need. It is best if you establish cooperation with someone from the medical community through personal connections, because otherwise it will not be easy to find an organization that will agree to work with you.

Determine the measured parameters of the state of health

Ideally, the data set obtained from the results of the senolytic tests includes information about (a) to what extent senescent cells were removed (b) to what extent the markers of aging were reversed. In reality, taking into account the current level of development of medical research, we have to compromise on both issues. After reading the literature and thinking about which indicators are likely to be affected by cell aging, based on current knowledge in this area, I chose the following analyses. For analysis, it is important that normal indicators vary greatly between 30 and 60 years, unless you are very old. This condition is not met for more available analyses than it might seem.

First of all, there are standard blood and urine tests. Comparing their indicators before and after taking the drug will most likely not show anything interesting, especially in people who are not yet 60, but they are inexpensive and can demonstrate that nothing terrible has happened. In addition, some blood test indicators are needed for other parts of the study. In particular, they can be used to detect signs of tumor lysis syndrome as a result of the destruction of senescent cells. The detection of a characteristic change in this indicator immediately after taking the senolytic indicates that something is happening, and this is useful information.

When considering the functioning of the liver, none of the indicators of a standard blood test does not seem particularly useful. The norm indicators vary little with age, but they strongly depend on the conditions and lifestyle. However, the results of cholescintigraphy vary significantly with age. This is an example of nuclear medicine using a radioactive isotope as a marker, and this procedure is worth it accordingly.

For kidney function, the desired indicator is the Rehberg–Tareev test. The result can be obtained in various ways. There is a direct and expensive method using radioactive isotopes, but there are also approximate methods based on data from a conventional blood test. The difference in the two approaches is described in a large amount of literature, for example, in a PDF from the National Kidney Association. Unfortunately, the evaluation method is not completely accurate in relation to the indicators of a healthy person. The MDRD formula should not be used, but you can try using the CKD-EPI formula.

Taking into account the available evidence in favor of the link between cell aging and vascular calcification, scanning and evaluation of calcium levels, at first glance, look interesting. This is so because the calcification level indicator is very difficult to reduce; this indicator increases gradually over the years and largely depends on lifestyle. Scanning for the level of calcification is simply a computed tomography, after which a semi-automatic analysis is performed, the result of which is an indicator on the Agatson scale or a specific calcification index (lesion-specific calcium score). Unfortunately, even in adulthood, a large proportion of people receive a zero grade, for example, more than half of 40- and 50-year-olds. If you are interested in these indicators, there is an online calculator created by one of the research groups in this field. All this makes the assessment of the level of calcification much less useful, taking into account also the cost of computed tomography. Perhaps it makes sense to try this technique only for people at least 70 and those who already know their indicator, and it is not zero.

Analyses of lung tissues suggest that removal of senescent cells can reverse the loss of tissue elasticity to some extent. It is worth looking at ways to measure the elasticity of the skin. The indicator can be obtained using cutometers or ballistometers sold on the market, while there are comments in scientific papers regarding the reliability of the results obtained. You may have to go not to a regular dermatological clinic, but to a plastic surgeon or to one of the dubious rejuvenation clinics. Perhaps more useful is an indirect method of measuring vascular elasticity based on the pulse wave velocity, which is easy to measure and the result of which changes significantly in adulthood. The question here, as in the case of other cardiovascular indicators, is to what extent normal indicators change due to primary factors of aging (including the accumulation of senescent cells) or secondary (weight gain, sedentary lifestyle). The use of this metric also checks the testing methodology itself, along with the testing of senolytics. For this reason, we cannot simply limit ourselves to choosing one type of analysis.

Another indicator of the cardiovascular system with suitable characteristics of age-related changes is heart rate variability. It is easy to measure. Note that the founders of the Palo Alto Prize chose heart rate variability as an indicator of biological age for interventions developed by competing teams.

Biomarkers of aging based on DNA methylation are now becoming an available means of research, although there is still no consensus on the methodology of their use. However, Osiris Green offers an estimate of biological age based on DNA methylation at reasonable prices. The price gives reason to consider such a method, although it requires no less checks than senolytics.

If it is possible to use personalized laboratory tests, you should pay attention to existing cellular aging tests or skin sample analysis, and both types of tests require a biopsy. In the first case, there are special kits for analysis and tests are accepted by the scientific community, but with uncertainty about the effect of biopsy on the aging of wound cells, which can make the results unreliable. In the latter case, the scientific article fully describes the methodology for its reproduction, but it remains to be guessed how useful it will be in practice. This is another case when the calibration of the research methodology is no less necessary than the analysis of the effectiveness of the senolytics themselves.

Selection of candidate substances for senolytics

First of all, let's exclude Dasanitib, Navitoclax and similar substances targeted at the Bcl-2 protein family from the list of candidates. These are relatively indiscriminate chemotherapy agents, and almost all other substances identified by the scientific community as potential senolytics are better, judging by the results of animal experiments – either have fewer side effects, or are more effective, or both. Of the remaining substances, it makes sense to try a combination, since some studies suggest that there is some synergistic effect between the candidates and the effectiveness of different senolytics against different types of senescent cells. In addition, scientific and corporate research at the initial stages rarely checks combinations of substances. Therefore, when using them, the chances of new interesting results increase.

Substances worthy of closer attention belong to two categories. The first is easy to obtain, similar to dietary supplements and relatively cheap drugs for oral administration, which are considered safe. Fisetin and Quercetin belong to this category, although there are doubts that the latter has any senolytic properties. The second is senolytics, discovered later, they are more difficult to obtain and use, and there is almost no data on their use in humans and safety, but they seem promising according to the results of recent studies. Piperlongumine and FOXO4-DRI belong to this category. In any case, you will need to familiarize yourself with the pharmacological data, with the studies where the drug was used, current ideas about its work, and decide whether you are ready to take the risk to take it. You will have to read scientific articles and assess your willingness to accept the associated risk. You should not blindly follow someone's advice, decide for yourself.

Determination of dose and regimen

Determining the dose for human trials requires reading a large amount of literature on animal studies to determine the most appropriate dose used in each case, which is often expressed in mg/kg, and multiplying it by weight. You will soon find that it is not easy to determine the dose for most senolytics, and you will have to make such decisions yourself. For example, Piperlongumine has so far been tested only on cell cultures regarding its effect on senescent cells. The literature describes its use as an analgesic with a dosage of 1-250 mg / kg, and for cancer suppression – 2.5-5 mg / kg, to increase the sensitivity of cancer cells to other drugs – 1 mg / kg and direct removal of cancer cells – 2.5 mg/ kg. In some cases, the assessment was carried out after a single application, in others, the application lasted several weeks.

Similarly, for Fisetin, there are no published animal studies regarding the effects on senescent cells. For other purposes, in recent years there has been information on pharmacokinetics for dosages of 10-250 mg/kg, in one study – 10-45 mg/kg twice a day and another for cancer suppression – 5 mg/kg twice for 2 weeks.

For Quercetin, you can see the original study of it as a senolytic, where researchers used a single dose of 50 mg / kg. There is very little data for FOXO4-DRI, with the exception of one recent article on its effects and another equally recent one on cancer. Access to both articles is paid and, unfortunately, the dosage data is not included in the main text of the original article, but is indicated in additional materials that I have not yet received. But I'll get them in time.

You need to understand that at the beginning you should try a very small dosage and gradually increase it up to the target. This is a reasonable precaution for many reasons. In some cases, senolytics are insufficiently tested on humans. Secondly, how can you be sure that the suppliers did everything correctly and that the substances were checked correctly? Moreover, when testing combinations that are not described anywhere, you may encounter unforeseen effects of mutual influence. Finally, if the substance works, and you have a lot of senescent cells that will collapse at the same time, there is a danger of tumor lysis syndrome. All these are very good reasons to increase the dosage gradually.

Of course, the mentioned drugs have different dangers, from Quercetin (sold in stores, and produced by many companies for years) to FOXO4-DRI (relatively new, little produced, delivered only by pre-order, without data on the effects on humans and only with a couple of articles on the effects on animals). When choosing a poison for yourself, be aware of the full level of risk to which you are exposing yourself.

Consider the method of use

Quercetin and Fisetin are drugs that can be placed in a jar, put on a shelf and stored for months. These are tablets intended for oral administration, which makes their use quite simple. Piperlongumine requires low storage temperatures and possibly powdered or incorporated into a mixture for ingestion. FOXO4-DRI is a short–lived protein that must be stored at low temperatures, then restored and injected - abdominal for mice, but probably intravenous is most suitable for humans. If you are familiar with how diabetics organize taking medications, the situation is very similar.

Injections are something in which it is desirable to enlist the help of a laboratory and a doctor, and not something that you need to do yourself. Note that given the current ridiculous war on drugs, which has been going on for more than three decades, caution should be exercised when receiving syringes for injection. This is another reason to organize everything in cooperation with a friendly laboratory and a doctor.

Identification of suppliers and products

The search for suppliers has different complexity for different senolitics. For Quercetin, you can cross the street, take a couple of packages from the nearest store and, subject to the reliability of the brand, safely skip the stage of self-checking the substance. You can also find an overview of the supplier's products online. Fisetin can also be ordered in the form of packages, but here the quantity and quality of suppliers is less known, so testing of the resulting substance should be considered.

For Piperlongumine, you will have to order the drug from a supplier and pay a significant amount – hundreds of dollars per dose, taking into account dosages from animal studies. For FOXO4-DRI, perhaps the best solution, given the small number of suppliers, is to order the synthesis of the drug from a company specializing in protein synthesis. This is expensive, and the assistance of the laboratory will be needed here. In both cases, suppliers are reluctant to supply the drug to someone who, in their opinion, will use it for human trials outside the framework of official research organizations.

Checking the substance

You will also need the help of a friendly laboratory to check the quality of drugs. Such quality control is not so simple and may require the use of special services, which are better obtained through a company working in this field than trying to order them yourself. It is important to make sure that you get what you pay for, both in order to avoid wasting money and time on independent experiments, and from the point of view of security. Even under optimal conditions, individual batches of substances that are not mass-produced may have poor quality.

Conducting an experiment

The first stage is to conduct all the analyses to determine the initial indicators. For many of them, such as a standard blood test, it makes sense to perform tests twice, possibly with an interval of several weeks, since the results may vary depending on external conditions. This is followed by taking the drug according to the scheme. Then there are two more stages of tests, one a few days after the end of the reception and one after a few months, and precisely because many indicators depend on lifestyle, it is important not to change the amount of exercise, diet and other conditions during this period.

After the job is done, all that remains is to present the entire study and publish it on the Internet for public access.

Choosing simple and complex paths

From this text it becomes clear that the complexity of conducting a successful experiment on oneself depends on the choice of a candidate for senolitics. The level of risk assumed is also of great importance. I chose senolytics partly to demonstrate this circumstance. In the simplest and safest experiment, you can use Fisetin and Quercetin and for the most part do without the services of specialized laboratories, relying only on the help of a doctor for blood tests, cardiovascular system and other medical tests. In this case, you can be fairly sure that the risk of negative developments is lower than for other drugs. Alas, these drugs are less likely to show good results compared to other candidates; it seems that there is no obvious optimal option in this matter.

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