A Review on Night Enhancement Eyedrops Using Chlorin e6

A Review on Night Enhancement Eyedrops Using Chlorin e6
Licina, G; Tibbetts, J

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Disclaimer
The authors of this paper are writing this review for research and informative purposes only. Increased light amplification may have adverse effects on the cellular structure of the eye if improperly used and the some of the materials used in this mixture should not be used on humans or animals.

Introduction
Chlorin e6 (Ce6) has been used for many years as a therapy agent in cancer treatment1,2.
However, in recent years other uses for ce6 have been found, the most notable in this case being its application into the conjuctival sac of the eye as a means of treating night blindness and improving the dim light vision of those with visual disturbances3. This preliminary study attempts to test the ability of a mixture containing Ce6 to improve the dim light vision of healthy adults.
In 2012 a patent was filed based in some part on the work of Washington et al4. The patent claims that a mixture can be made which, when applied to the eye, will absorb to the retina and act to increase vision in low light. The mixture put forth in the patent is a simple combination of Ce6 and insulin in saline. It is mentioned in the same, that dimethlysulfoxide (DMSO) can be used in place of the insulin. We propose a combination of the two could lead to the most noted effects. For testing purposes, the mixture from the patent (Ce6, Saline, Insulin) was used with the addition of DMSO for increased permeability.

Material Background
Ce6 is a tetrapyrolle and a chlorophyll analog. As mentioned, it has historically been used as a photosensitizer in laser assisted cancer remediation. The light amplification properties of the Ce6 are used to use the energy from a low power light source to destroy cancerous cells with literal laser precision. The reaction creates oxygen species which induce apoptosis in tumor cells. This lead to the concerns about the mixture, as it would be possible that bright or even ambient daylight’s amplified effect in the eye may harm the cells, potentially causing permanent damage.
The function of the insulin is not expressly mentioned in the patent or the journals papers. It has been shown that insulin downregulates the ABCG2 mediated transport pathway5. With ABCG2 downregulated, greater absorption is shown for photosensitizers like Ce66. In the case of this solution, the insulin is used to allow absorption of the Ce6 into the chamber of the eye.
DMSO is used in cell preservation and in medication application. It’s primary ability, in this scope, is to cause increased permeability of the cellular membrane, allowing for free passage for any chemicals that come into contact with the dosed area. While anecdotal reports of healing abilities or “tasting” lemon juice through ones skin abound on the internet, the high risk of cellular toxicity from outside contaminants being absorbed through the skin make this chemical something that should only be handled with caution.

Procedure
The Ce6 (Frontier Scientific, CAS: 19660-77-6 ), was found to be a fine black powder which clung to all surfaces. To make manipulating the chemical easier, a large batch of the total solution was made and then aliqouted into separate containers for storage.
200mg of Ce6 was mixed with 400 units (4ml) of insulin. To this was added 5.38ml of sterile saline solution (0.9% sodium chloride). The mixture was sonicated briefly (30 seconds) to allow for proper dispersal of the powder into saturated solution and then 625μl of DMSO (Amresco) was added. The solution was sealed with parafilm and sonicated for 150 seconds. The resulting liquid was thin and black in color. Solution was kept in glass aliqouts wrapped in foil at 20°C.

For the application, the subject rested supine and his eyes were flushed with saline to remove any micro-debris or contaminants that might be present. Eyes were pinned open with a small speculum to remove the potential for blinking, which may force excess liquid out before it had a chance to absorb. Ce6 solution was added to the conjunctival sac via micropippette at 3 doses of 50μl into each eye. After each application, pressure was applied to the canthus to stop liquid from moving from the eye to the nasal region. Each dose was allowed to absorb between reloading the pippette, with the black color disappearing after only a few seconds.
After application was complete, the speculum was removed and black sclera lenses were placed into each eye to reduce the potential light entering the eye. Black sunglasses were then worn during all but testing, to ensure increased low light conditions and reduce the potential for bright light exposure.

Testing
The Ce6 solution has been shown to work in as little as one hour, with the effects lasting for “many hours” afterwards3. After 2 hours of adjustment, the subject and 4 controls were taken to a darkened area and subjected to testing. Three forms of subjective testing were performed. These consisted of symbol recognition by distance, symbol recognition on varying background colors at a static distance, and the ability to identify moving subjects in a varied background at varied distances. Symbol recognition consisted of placing a collection of objects with markings on them (numbers, letters, shapes). Subjects were then asked to identify the markings, each viewing the objects from the same location at a distance of 10 meters. The markings were not made prior to the moment of testing.
For subject recognition, individuals went moved in a small grove of trees. They were allowed to chose their own location independently. Distances ranged from 25 to 50 meters from observation point and trees and brush were used for “blending”. Locations were chosen without being observed by the test subjects. The Ce6 subject and controls were handed a laser pointer and asked to identify the location of the people in the grove. After testing the Ce6 subject replaced the sunglasses which were not removed until sleep. Eyesight in the morning seemed to have returned to normal and as of 20 days, there have been no noticeable effects.

The Ce6 subject consistently recognized symbols that did not seem to be visible to the controls. The Ce6 subject identified the distant figures 100% of the time, with the controls showing a 33% identification rate.

Discussion
It is noted that more testing will need to be done on this particular project. Current testing done was subjective in nature. A Ganzfeld stimulator and electroretinigraph will be used to measure the actual amount of electrical stimulation increase from the eye, giving a hard quantifiable number to the degree of amplification. It is also possible to test which ranges of vision are being amplified as well. However, given the current results and the previous body of work on the technique, it seems fair to say that this technique is successful in it claims for low light amplification in the human eye. These findings are subjective experiences. Subject experienced no adverse effects following administration. Preliminary testing seems to indicate this increase in dim light vision to be occurring. Further testing is need to confirm and measure the degree of improvement in health subjects.

Being able to access the information put forth in journals and patents is extremely important for future scientists to be able to work with and build from the knowledge that we have currently. Moreover, it is extremely important for clear methods to be available for any researcher who desires to review a scientific paper. The last year (2014) has shown more scientific journal articles rescinded than any year previously. Citizen scientists and “DIY biologists” are under no pressure to reach or hold a position of tenure and often do not have the need to produce for monetary reasons. It is possible that this will allow for less bias in publishing and a more open release of work due to the lack of external motivators. By making information accessible, one can pre-empt “scooping” and instead focus on collaboration. During this research, we feel we were fortunate to be operating from just such a position. The disadvantage however was a decreased availability of access to many of the tools that would allow us to verify our findings quickly and easily. Ce6 administered as described in this paper in the dosages described have so far been without any adverse effects and show great potential to enhance the vision of healthy adults in dim light situations. Further studies should be performed in order to measure the effects of this ce6 solution objectively.

References

1 – Bachor R, Scholz M, Shea C, Hasan T. Mechanism of photosensitization by microsphere-bound chlorin e6 in human bladder carcinoma cells. Cancer Research, 1991, 51:4410-4414.

2 – Akhlynina T, Rosenkranz A, Jans D, Sobolev A. Insulin-mediated Intracellular Targeting Enhances the Photodynamic Activity of Chlorin e6. Cancer Research, 1995, 55:1014-1019
3 – Shantha, T, inventor, assignee. Methods to Enhance Night Vision and Treatment of Night Blindness. 21 June 2012. Patent 20120157377.

4 – Washington I, Zhou J, Jockush S, Turro N, Nakanishi K, Sparrow J. Chlorophyll Derivatives as a Visual Pigment for Super Vision in the Red. Photochem. Photobiol., 2007, 6:775-779, DOI: 10.1039/b618104j

5 – Liu X, Jing X, Jin S, Li Y, Liu L, Yu Y, Liu X, Xie L. Insulin suppresses the expression and function of breast cancer resistance protein in primary cultures of rat brain microvessel endothelial cells. Pharmacological Reports, 2011, 63:487-493

6 – Robey R, Steadman K, Polgar O, Bates S. ABCG2-Mediated Transport of Photosensitizers: Potential Impact on Photodynamic Therapy, Cancer Biology & Therapy, 2005, 4:2, 195-202, DOI: 10.4161/cbt.4.2.1440

82 thoughts on “A Review on Night Enhancement Eyedrops Using Chlorin e6”

  1. I would would be weary of using Chlorin E6 in your eyes. The role of Ce6 in chemotherapy is as a photosensitizer so you’d expect that it would produce reactive oxygen species when exposed to light. I don’t know that generating ROS in your retina’s is such a great idea. Also, there is no explanation as to WHY one would include insulin in the mix. What does insulin do? Why is it necessary? DMSO alone should do the trick for delivering Ce6 on its own.

      1. Ah, OK. Missed that part. What is the mechanism of Ce6 light sensing enhancement? If the molecular mechanism was available, then one might find compounds that would provide the same effect without the potential danger of ROS. Has any one tried heme derivatives such as bilin or even the corrin ring system from cobalamins? They are structurally similar and, to my knowledge, do not function as photosensitizers and have great quantum efficiencies. I am a PhD candidate in a lab that does long wavelength photochemistry and develops light-controlled tools for biological processes. If you’d ever like to chat more about this stuff let me know!

        1. Wow! yeah, nobody has tried any of this stuff. It’s not really monetizable and therefore no lab really cares, especially to try it on people. We just found it and decided to run with it. We have the background and a allergy to doing anything that people actually pay us for, evidently.

          Please email us at projects@scienceforthemasses.org. We would love to talk more about tweaks to this system and more 😀

          1. By not really monetizable, you mean that 1. the military doesn’t want it (they do, i assure you), 2. The general population doesn’t want it (they do, I assure you), and 3. Everyone else doesn’t want it (they do, I assure you)?

  2. I would also like to volunteer as a test subject in any future studies. You guys are doing amazing work, and I would love the opportunity to help you make history!

    1. are you reallly certain you would like to be a human guinea pig?
      how much money would they pay you, anyway, for undergoings their tests?
      no amount of money would ever restore your vision if, heaven forbid, anything awful were to happen.

  3. Including antioxidants such as trolox (vitamin E derivative) and pyranose oxidase/catalase (oxygen scavengers) may enhance the chromophores lifetime while also reducing damage from ROS buildup. This is what we do for single molecule fluorescence microscopy and has been shown to work with Ce6 utilizing cancer therapies. You’d have to play around with concentrations though as I’ve never seen these compounds delivered to the eye.

    http://www.ncbi.nlm.nih.gov/pubmed/17822943

    1. I think you make a good point here regarding antioxidants, but I would be surprised if the enzymatic scavengers would actually help. The likelihood that they would be absorbed into the aqueous humor and actually make it to the site of action is very low as there would be multiple membranes to cross and the ROS produced generally act very close to where they are produced. Therefore, if the Ce6 was acting inside of cells, then the enzyme’s would do very little to help since they would essentially be floating on top of the cornea. I think that antioxidants such as trolox and potentially various carotenoids would work best. You could probably add retinoic acid itself and do a good job of helping quench ROS and maybe even help boost their vision a little bit.

    1. Dino – Very sorry to hear about about your lasik experience. I’ve worn thick glasses virtually all my life, but have specifically chosen not to go with the lasik procedure. The track record for various versions of the lasik procedure is somewhat spotty at best. I worked with one of the world’s first attempts at commercially developing this (Phoenix Laser Systems) in the mid 1980s. To this very day I’m still very cautious in suggesting this procedure to anyone.

  4. The results you obtained would be much more impressive if you would have taken the time to make this a double-blind study, considering you are using subjective results. I understand that people may not have been lining up to have this concoction dropped onto their eyes.

    From an Ophthalmology standpoint, Verteporfin (Visudyne) has been used intravenously to photo-sensitize the retina to perform Photodynamic Therapy to treat wet macular degeneration, Diabetic macular edema, and choroidal tumors for quite some time. This drug has a very similar wavelength absorption spectrum as the Chlorin e6 you are using. This absorption takes place in the upper 600nm wavelength, the limits of the visible spectrum (red).

    This drug flows through the choroidal vasculature and when stimulated with the laser releases ROS that causes fibrosis of leaking vessels. For the next 3 days the patient is asked to avoid direct sunlight since some of the drug eventually gets into the actual retina which is not where you want ROS being set off (as this is a suspected cause of vision loss, macular degeneration, and choroidal neovascularization).

    Now your taking a “drug” with similar wavelength absorption properties and purposely trying to get it directly into the retina. Imagine millions of tiny oxygen bombs going off in your rods, cones, and retinal pigment epithelium. There is a reason the prior studies you have referenced involve mice models and not human subjects. You comment that “so far” there has been no adverse effects, but that is on a gross subjective scale. Your not using OCT’s or fluorescein angiography to monitor for potential retinal changes on a cellular level.

    I know its not your fault that this has popped up all over the internet as “injecting night vision into your eyes” but it is severely misleading and I just pray people are not running around trying to obtain some Chlorin e6 and Lantus to go mix up and squirt in their eyes to follow in your foot steps of potential irreversible retinal damage.

    1. I am totally disgusted by this conduct. Research experiments in animals require protocols approved by ethic committees. Research on human subjects also required FDA (in the US) and Ethic Committees review. I am not able to find Tibbetts and Licina’s credentials, particularly in post-doc advance degrees. Furthermore, the paper was just a write up and it has not been published in a reputable journal. I’ll gather all information and submit to FDA for false advertisement, fault expectations, and putting subjects at risk without any scientific, medical nor ethical review.

      1. It’s a blog post. That’s it. We have no control over sensational news stories.
        Furthermore, there are no restrictions on if I want to put something in my eye. I’m not making you do it. Research on human test subjects requires all sorts of paperwork, research on myself requires none. We’re not selling anything so it’s not false advertisement, we are using reputable sources so the expectations that we put forth are in line with the research that has already been published, and the subject I put at risk was myself. You aren’t going to get me to sue myself. Who is the harmed party?
        Relax.
        We’re terribly sorry that you didn’t find a post doc degree attached to our names which is, I am assuming, the magic pass that allows people to play around in a lab. However, you can google me in scholar and find a few papers with my name on them.

        Did you even read the paper, follow the references, do any research of your own?

        1. Hello, since I would like to avoid any “sensational news story”, I would love to be able to ask you a few questions. Fact checking, you know. Maybe by email since you live on the West Coast, that’s way behind France hourwise. I am a French journalist. Thanks.

      2. Oh, for shits sake, cut the elitist crap.

        Galileo stole corpses 500 years ago because he lacked “credentials”.

        And today? “Ethicists” are something only large organizations can afford. They are inexorably biased in favor of their employers (at least minimally), and the big organization, at the CEO/CTO/CFO level could really give a shit about the ethics if the money is big enough.

        The only thing these guys would gain by going through an “ethics review” beyond what they already did themselves, is the chance someone else would steal their work and use their greater access to capital to shove these guys out.

        As to the real risk…the very same liberals objecting to the “Informed Consent” of the test subject on “ethical grounds” have no problem with people using mind and DNA-altering drugs (marajuana, ecstacy, ghb, vapor inhalants, cocaine, heroin, LSD etc.) for personal enjoyment, even though the long term affects of such use are generally negative, including defects in offspring.

        Night vision eye drops? BFD. You’re just pissed you didn’t think of it first and want to throw logs across the road they are following to slow their advance.

        There will be plenty of testing and oversight before a commercial product is available.

        How about applauding these guys for their initative, ambition, and courage instead….takes balls to let someone drip such a concoction in your eyes! Bravo to the subject!

        1. Takes balls indeed. Eyeballs.. I applaud these guys for taking the first steps down this road in science. If it wasn’t for people like these guys/pioneers, science would have been stifled long ago by the bureaucrats.

  5. No monetization? Are you kidding?:) How about militarization? That’s really useful, if we would know, how to desaktivate this stuff fast. To not be blinded. Can somebody try it out of laboratory? At home? Or it’s dangerous?

      1. Did the “controls” also wear sunglasses inside? How long were they in darkness before testing began? The treated subject would have already adjusted to darkness because of sunglasses.

    1. Did you follow up on the resources we mentioned in the write up? I mean, there is a patent for this very thing that has existed for years as well a few papers describing the mechanism in murine models…

        1. It was subtle. Things that were dark became dim, things that you needed to squint to see became something you could just look at. We waited 2 hrs before we checked if it worked or not. The figures were made with black marker on balloons (it’s what we had) and there were (sadly) no special colours.

  6. I would like to try bing a subject. My job revolves around alot of night vision and field equipment using ir capability might also be interesting to use with the night vision and can give some feedback to it.

  7. I have retina pigmentosa with related poor night vision and progressively worse fields of vision as light levels drop. I’d be very interested in participating in any trials in the UK.

    1. They totally are.
      Sorry everyone, but despite how thrilled people are about this, the effects are way more subtle than the media would leave you to believe.
      This is just enhanced night vision. Dark becomes dim, it’s not all Riddick up in here.

  8. Interesting experiment. I suggest getting some amateur or pro astronomers involved for the next round of experiments. We know the brightness of stars to high accuracy, so the ability to see fainer stars would accurately show the amplification available. Objective as all hell.

    1. I was asking myself the same. If you have any information of this studies helping retinosis picmentaraia please let me know.

  9. Just heard in on TV and read it in the news. Great idea and work. Keep it up, I hope you guys can develop and improve it.

  10. I have experienced similar results using Escitalopram. I experience intense night vision, when I first started taking in the dark(near darkness) would be green but visible. After time, night vision would be superior, but more of a white(ish). I know that escitalopram interacts with the Gaba receptors of the retina through online research.

  11. How have subjects with retinal abnormalities reacted when exposed to Ce6? Has Ce6 been effective in improving night vision in subjects with rentinitis pigmentosa?

    1. Unfortunately, it’s hard to say. There has been no testing on this matter in humans.
      While we haven’t done any work on this ourselves, there are some sources that point to this.

    1. Night vision in an individual can be enhanced through practice, just by going out at night into unlit areas like jungle or forest, where the ambient light is very minimal to none. Studies have shown that back in the day when soldiers, especially infantry, used to constantly train at night, in which not everybody had night vision optics for every soldier, their night vision actually improved, as a result of the human body’s natural ability to adapt to it’s environment. However not everybody is the same so some people cannot adapt, most likely due to genetics. Again. however most soldiers that did spend huge amounts of time training at night had experienced night vision adaptation, basically having good natural night vision capability. I cannot say that I was born with good night vision capabilities but I can tell you that as a former soldier I have spent more time training at night than at any other time of day and that I have excellent natural night vision capabilities. I am older now and I still have good night vision but it does take longer for my eyes to adjust, probably due to age. Science is awesome and fully support what these guys are doing, but do not underestimate the human body’s ability to adapt. It’s exactly how we made it this far without becoming extinct as a species. Nature can force us to adapt as it always has, however we are the only species that can create our own conditions to force our own adaptation to our environment. Pretty amazing, right?

    1. I do nondestructive inspection (always in the dark) I think this would have an added attraction to that. I will email you. Thanks great job, remember many naysayers got to the top because they showed the company why it wouldn’t work. Thus the company didn’t fund it, and this it was “proved” it couldn’t. ….

  12. DEAR ANYONE WHO WILL LISTEN: instead of experimenting on rats, or any other living creature, could they not experiment on vegetable tissue … and come up with the same results. repeat; same results!!

    1. The solution works by being absorbed into the eye. Applying it to a contact lens would not only hinder that process, but also create a variable in how much solution actually reaches the eye, making the results more unpredictable.

  13. Hello, I know its kinda late to ask this, but I still don’t understand how this eyedrops works? Is it make a membrane in our retina or is it solved in the retina and increase the photosensitivity of the rods? I’m sorry i’m a senior high school student so I can’t understand this clearly. And fyi your experiment was in my biology test!

    1. Ahahaha! That’s awesome!
      The Ce6 is absorbed into the eye and then you can kind of just imagine it as a free floating molecule. Of course, we don’t know if it adhered or not, it just makes a bit more sense that it’s in solution (the solution being your eye jelly).

      The prevailing theory is that the Ce6 absorbs the light coming in and kicks it out at an altered level. So your eye doesn’t change, more like the light coming into the eye is changed.

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