F. Diazirinikovic, H. Doof and C. Baschovcinovic
Abstract: We enrich the world with yet another method to kill cancer cells.
Specific: We report the designed, synthesis and activity of a novel photoactivatable glucose derivative: a diazirine- conjugated benzoylated sugar (DiaBeTeS). We show that when cancer cells are treated with diabetes and subsequently irradiated with ultra-high doses of UV, they die. This toxicity was highly significant (p = 0.043) and was absent when cells were treated with either UV-light or DiaBeTes alone, as shown by p-values that were just above 0.05. The potency of our compound was comparable to conventional cell-killing agents, such as ethanol, chlorine and the music of Cardi B.
Introduction
If you gave 100 people the choice between sugar and cancer, 99 people would pick sugar, and one person would say “por qué no los dos” out of habit. What fewer people realise is that if cancer could talk, it would pick sugar too.1 Cancer cells, like any others, require chemical energy in the form of glucose, and this makes them vulnerable. We set out to exploit this weakness by developing a cytotoxic analogue of glucose. The only flaw in this plan is that healthy cells also need glucose, and as such without a way to aim our molecule at a tumour, we might as well be treating cancer with cyanide.
To balance the desired properties of toxicity and specificity, we selected the diazirine functional group as the reactive handle for our glucose analogue. This energetic heterocycle expels N2 on exposure to UV-light, generating a highly energetic carbene which rapidly reacts with whatever it can get its HOMOs on. In short, we predicted that glucose functionalised with a diaziridine could serve as a molecular claymore by blowing the face off a cancer cell when triggered by UV.
Results and Discussion
The detailed syntheses of our molecules are described in the supporting information of a follow-up paper that we haven’t written yet. The most important points are that we had to protect all the hydroxy groups of glucose (1) with benzyl groups before we installed the diaziridine, and that once it was installed, we couldn’t get the benzyl groups off (2). This actually transpired to be beneficial, as the benzyl groups vastly increased the metabolic stability of our compound, at some small cost to its solubility (data to be disclosed in the second follow-up paper). Finally, we synthesised an alkynylated analogue of DiaBeTeS (3), because that’s what everybody else does.
We then tested the ability of DiaBeTeS to kill cells. We incubated a cancer cell line with 100 mM DiaBeTeS for 24 hours, and subsequently irradiated the cells with UV light of 180 nm for another 24 hours, with a total dose of approximately 850 mSv. We then measured the viability of the cells with an MTT assay (Figure 2A). Treatment with DiaBeTeS or UV alone clearly did not kill the cells, as indicated by non-significant p-values of 0.063 and 0.081, respectively. However, a synergistic effect was observed when DiaBeTeS -treated cells were irradiated, killing the cells instantly (p-value 0.043). Clearly these results show that DiaBeTeS and UV are only toxic when they are combined, making this is very selective and safe method to kill cells.
We then compared our method to other commonly used methods of killing cells.2 Treatment with DiaBeTeS & UV was almost as effective as treatment with 70% ethanol or chlorine (Figure 2B). This result demonstrates the potency of our novel method to kill cells with great selectivity and makes it attractive for further development. Naturally, we filed a patent protecting this technique to prevent any further development.
Given that we’d already prepared an analogue of our drug with an alkyne attached, it seemed logical to determine the target proteins of DiaBeTeS by performing a pull-down assay with mass-spectrometric analysis.2 For the data thus acquired (you guessed it, third follow-up paper), we identified a number of candidate proteins (Table 1).
First of all, DiaBeTes seems to act partially through the inhibition of trypsin by covalently modifying this enzyme. Secondly, DiaBeTes effectively targeted keratin, making it not
only an effective anti-cancer drug, but also a potential cure for, or instigator of, baldness. Thirdly, by being able to target chicken egg avidin, this molecule could serve as a molecular tool to separate egg white from the yolk. Lastly, DiaBeTeS targets several structural proteins like actin and tubulin. The slightly worrying implication of this finding is that DiaBeTeS may have side-effects, such as generating funny-looking cells, and maybe even funny-looking patients.
Taken together, these results indicate that DiaBeTeS does not have a singular molecular target, but exhibits broad-spectrum cytotoxicity through multiple pathways. In a set of preliminary experiments, we also found that our compound renders cells unable to excrete pro-inflammatory cytokines. We therefore envision that it might serve as a potent anti-inflammatory therapy for the treatment of neurodegenerative disease, auto- immunity, covid-19 and ageing.
Conclusion
Given the astounding results disclosed above, the logical next step is to proceed directly to clinical trials. This will begin as soon as we source enough willing volunteers, and when the FDA stops pestering us about off-target activity. There is a slight chance that if a person taking DiaBeTeS gets into a sunbed, their entire epidermis will explode, but we’ll cross that bridge when we come to it.
Author Contributions
F.D. performed all the work. H.D. is just on this paper for political reasons. C.B. funded the work.
Author Contributions
H.D. is the owner, stock holder, employee and only board member of Doof Laboratories Incorporations. H.D. has filed patents to commercialize the methodology described herein for curing cancer, industrial scale egg separation and world domination. C.B. is the owner of a company that produces cookies incorporating DiaBeTeS.
Acknowledgements
We would like to acknowledge Perry the platypus for continuous support and feedback on this manuscript.
Notes and References
“Jump up jump up and pull down: an MS-FUNKY assay for protein conjugation” T. Boy, C. Hill, 1992, J. Mus. Chem., 6(8), 1:30.
Note: if cancer could talk, it would almost certainly be on twitter.