In 2015, indysci’s Project Marilyn began scientific research.
In April, indysci began renting chemistry space from QB3, and I obtained a materials transfer agreement allowing work using the 9DS-producing strains to begin. The company Happilabs was contracted to do the initial lab setup, resulting in major savings. To save on rent, I chose to grow the bacteria at a home “biology lab” in the garage instead of at QB3. I began to practice growing sibiromycin, which is less complex and less risky than growing 9DS, in order to develop the technique and iron out any bugs in the process.
By June, I achieved first production of sibiromycin, and over a few months became robust enough to confidently work on 9DS. After a brief snafu where I discovered the wrong compound was being fed to the bacteria, I achieved first production of 9DS in October.
Meanwhile, working in the chemistry lab, I discovered the “old” procedure for isolating and purifying 9DS was inefficient. From September through December, I tested various alternative steps were tested to improve yield. One of the most promising changes is the use of a lyophilizer to freeze-dry the product in the initial stages, resulting in a whopping 5-10x improvement. That means that the process of producing 9DS could be 5-10 times faster. This lyophilizer was purchased second hand at a steep discount and refurbished to perform as well as a new device!
Like the sibiromycin strains, I found it necessary to improve the 9DS production strains, as they were unreliable at first. Each round, I took from the best performing flask, inoculating the next round with this sample. I believe that over time this engages artificial selection to create progressively hardier strains. At the very end of the 2015, I set up an experiment to test the reliability of 9DS production, and at the beginning of 2016 I found that the culture from that batch achieved 4/4 positive production – a milestone meaning that scaling up of production is going to be smoother from here on. At the beginning of Project Marilyn, I predicted that establishing good microbial cultures would be the most risky and difficult part of the process, and so it feels very good to have completed this.
What to expect in 2016:
As production ramps up, I will be purifying more of the compound. Already I’m accumulating milligrams of material, isolated from everything else. Once the procedure for purification becomes very efficient, expect to see characterization of the compound using NMR (“MRI for chemicals”) which will also give an idea of how pure the sample is. Finally, when a sufficient amount of 9DS is produced, the material will be submitted to the big experiment: Will it cure cancer in mice?
As a reminder, the donation of >10 bitcoin unlocked a goal to test 9DS derivatives to see if they are also biologically active. Now that the strains are performing consistently, I will begin work on this goal. If any of these derivatives look promising, the next step will be to bring these compounds through to preliminary petri dish and mouse testing as well.
To better follow the progress of Project Marilyn, you can watch the Facebook page, where updates are more frequent: http://facebook.com/