1. Summary

BioRegenerative Sciences was formed in 2009 to translate the discoveries of Dr. Greg Maguire while a professor at the University of California, San Diego School of Medicine. Those early studies at UCSD in the 1990s helped establish that adult stem cells release molecules into damaged tissue to induce tissue repair and regeneration, and that two (2) or more types of stem cells were involved, each type releasing a different pool of molecules to effectuate a synergistic healing response. The combination of molecules from a stem cell is called SRM (stem cell released molecules), and the SRM from two or more stem cell types is called S2RM. Maguire and Friedman developed these ideas into a patent-pending platform technology for the commercialization of a number of therapeutics, including the BRS eye drops, called I-Ease, for Dry Eye, and another eye drop, called Lens-Ease, for Cataract. These products contain the S2RM® molecules, and not any cells or tissues. Our approach has been described as “stem cell therapy without the cells” (Maguire 2013), leading to a type of therapeutic called a “systems therapeutic” (Maguire, 2014). Both products have been shown to be highly safe and efficacious in animal and human testing. Furthermore, while many new drugs/biologics brought to market recently were shown to have positive clinical risk benefit, many of the new drugs currently awaiting pricing from reimbursement authorities have not addressed relative cost benefit [1]. In contradistinction, the efficacy and the cost of the BRS products both display great relative superiority to comparative products on the market.

2. Background [Stem Cells, Stem Cell Products]

Adult stem cells in the human body are present in most tissues, including the eye, serving to regenerate and repair the tissue, quell pain, and modulate the immune response. Approximately eighty percent (80%) or more of the healing capacity of adult stem cells derives from their release of multiple molecules, including growth factors, cytokines, interleukins, antioxidants, exosomes, microRNA, and chaperone proteins, into the damaged tissue resulting in the orchestration of a healing response. The unique nature of the S2RM® technology centers on the identification, selection, culture, and stimulation of the appropriate stem cells, and then the capture of those molecules released from the stem cells. In this manner, proprietary and patent-pending technology is used in the BioRegenerative Science (BRS) laboratory to capture what the stem cells normally release in the body. This is a systems based approach where, instead of a reductionist approach using one or two molecules for treatment, all of the molecules from the stem cells act together as they normally do in the body in order to deliver the emergent properties of the system of all molecules. Further, BRS uses a technology that mimics the manner in which the body heals itself whereby two or more types of stem cells become resident at the site of injury, and the two or more types of stem cells release their pools of molecules into the tissue in concert, producing an emergent and powerful, synergistic healing effect. The overall effects of S2RM technology are twofold: 1. Mimicking the effects of SRM on those cells surrounding the stem cells, and 2. Enhancing the effects of the endogenous stem cells themselves.

The S2RM® technology, exclusively used by BioRegenerative Sciences, has the advantage compared to other stem cell technologies of capturing a full and natural complement, or “system,” of known molecules. This full complement of captured molecules from stem cells are subsequently injected into, or topically applied to, the patient in a defined and precise spatiotemporal dosing schedule to begin the systems-based regenerative process. The known dosing parameters for S2RM are in contradistinction to that for stem cell-based therapies where the cells cannot easily be controlled once they are removed from the laboratory setting and administered to the body.

Examples of topical S2RMTM Technology include eye drops for dry eye therapy, and for cataract.

3. The S2RM® Technology for Dry Eye and Cataract

A. Dry Eye

Tear fluid forms a tear film over the cornea and the conjunctiva of the eye. Dry Eye is associated with a reduction in the quantity and/or quality of the tear film. The tear film contains aqueous solution, a broad spectrum of fatty acids [2], and a multitude of signaling molecules, including SRM [3]. The composition of the tear film is significantly altered in dry eye, including a down regulation of many proteins as analyzed by proteomics of 386 protein types in human dry eye patients [4]. The source of these proteins in the tear film, as well as the source of other signaling molecules, is not well understood but likely from multiple cell types, including stem cells, in the various ocular tissues. The constant bathing of the anterior ocular tissue with SRM is thought to be important for maintaining the cornea and conjunctiva. The BRS approach to restoring the tear film in dry eye is to replace the missing signaling molecules though an application of SRM to the corneal surface. The BRS product is called I-Ease, a sterile ophthalmic solution that is dropped into the eye and supplies S2RM® to the anterior surface of the eye to bathe the cornea and the conjunctiva in all of the molecules needed to maintain and heal tissue.

The mechanism of action of I-Ease is believed to: 1. increase the anti-oxidant capacity of the cornea, 2. recondition the stem cell niche of the limbus and 3.mimic the release of SRM from the resident stem cells in the stem cell niche of the limbus and secreting glands of the eye, including the meibomian gland.

B. Cataract

As we age, the continued incidence of photons on the lens tissue, and the consequent oxidation of the tissue, along with other aging mechanisms, including stem cell dysfunction in the lens capsule and epithelium [5,6], degrades the lens tissue and results in opacification. While little is known about the exact causes of cataract, we know that, for example, the lens capsule is a reservoir for growth factors [5] to maintain the tissue, and that a high antioxidant capacity within the lens is needed to buffer the len’s constant light exposure. The BRS approach to restoring lens function and to preclude or reduce cataract formation is to replace the missing signaling molecules though an application of SRM and antioxidants to the lens. The BRS product is called Lens-Ease, a sterile ophthalmic solution that is dropped into the eye and supplies S2RM® and added antioxidants to the lens, thus bathing the lens in all of the molecules needed to maintain, protect, and heal tissue.

The mechanism of action of Lens-Ease is believed to: 1. increase the anti-oxidant capacity of the lens, 2. recondition the stem cell niche of the lens capsule and 3. mimic the release of SRM from the resident stem cells in the lens capsule stem cell niche.

Figure 1. Schematic of the eye showing the Cornea, and the Lens.

Figure 2. Location of possible stem cell niches partially responsible for production of SRM bathing the cornea.

4. Results for Dry Eye and Cataract

A. Dry Eye I-Ease

Over 100 patients successfully using the product for severe dry eye, post-LASIK irritation, Fuch’s corneal dystrophy, corneal ulcer, and post-transplant irritation. Ophthalmologists in North and South America carry the product in their offices for sale to patients, and the product is being recommended by Dr. Anne Coleman, MD, PhD., faculty at Jules Stein Eye Institute, UCLA School of Medicine, and Dr. Colleen Grace, M.D. at the University of Michigan, Henry Ford Hospital.

Figure 3. Before and after picture of dry eye in canine. I-Ease smoothes the corneal surface and reduces irritation after one week of application.

Testimonial:

Dear BRS,

I began wearing contact lenses when I was a senior in high school. I wore contacts daily for a period of about four years with no problems at all. I regularly replaced my contacts and followed each of my doctor’s instructions. After some time, I began to slip up and occasionally fell asleep with my contacts in. I began to notice a minor burning sensation and irritation when wearing contacts, which progressively worsened until I was unable to wear contacts at all.

I thought that by taking some time off, wearing my glasses and allowing my eyes to heal, that I might again be able t wear contact lenses. Turns out that I was wrong, my eyes did not heal on their own, not even after 4 years of wearing glasses. I tried several treatments as directed by my doctor, but nothing worked. I had all but given up on ever wearing contact lenses again.

Then, I met Peter and Greg from BioRegenerative Sciences, who gave me a free sample of I-Ease. After using I-Ease twice a day for about a week, I finally was able to wear my contact lenses.

I now wear contacts every day, and continue to use I-Ease for maintaining healthy eyes. I would recommend I-Ease to anyone who has trouble wearing contact lenses due to irritability.

Thanks BioRegenerative Sciences!

Erika Csaszi

San Diego, CA

B. L- Ease

Pilot studies suggest L-Ease is powerful in preventing and reducing cataract formation. Thus far 15 people are using the L-Ease with positive results. Several patients are being followed by ophthalmologists or other health care providers. Imaging shows the cataract diminishing in size over a period of several months, and patients report fewer visual disturbances after 1-4 months of usage. Additional animal and human studies are needed.

Testimonial:

Two years ago, at a routine eye exam, I was told that I had small cataracts developing in both eyes, nothing requiring surgical intervention, but to be watched. Given the extensive sun exposure in the past, combined with aging, this was not a huge surprise. The next year I was told the same thing, small, but to be watched. I was noticing one difference in my vision – an increased amount of “starring” around lights at night, which was very annoying, especially as I sometimes drive long distances at night. About 6 months after the last exam I started using the L-Ease, initially one drop a day, sometimes one drop two times a day if I remembered it. The drops have not caused any irritation or difficulty with my vision after using them, no side effects that I have noticed at all and I have been able to continue using my gas permeable contacts as usual. The one difference that I experienced is a decrease in the “starring” after using the drops for about 3 months. This was a notable difference for me and continues to be markedly improved. Last week I had my yearly eye exam. The description of my cataracts when I inquired was “trace”, “minimal”. It was also interesting that I have a slight improvement in my astigmatism in the Right eye. I have had significant astigmatism in both eyes since grade school which has only worsened, never improved at all. I’ve not had a Lasik procedure or used any medications or interventions other than the L-Ease, which would account for the changes that I have chronicled. I will continue to use the L-Ease, as I feel I can only benefit. I’ve had an increase in quality of my night vision and quantitative improvement in my eye health instead of a decrease in eye health which accompanies aging. Great drops!

Rita Friedman, RN, PhD, JD

Medical and Scientific Research Committee

BioRegenerative Sciences

5. Platform Production Technology

The BRS “Intelligent Molecules” Expression System, described in our paper (Maguire et al, 2013), employs a proprietary manufacturing process that allows the production of a wide variety of soluble and properly folded proteins and signaling molecules from multiple stem cell types important to immune modulation, and tissue repair and regeneration. The advantages of the process are in the production of biologically active proteins, exosomes, and signaling molecules, developing a “systems therapeutic” yielding a combination of many molecules, that act at multiple targets, resulting in a synergistic therapeutic with emergent therapeutic value. Furthermore, the production of the SRM (stem cell released molecules) does not require downstream solubilization, refolding, or other processes. Additionally, the process offers reduced purification requirements and lower production costs than other pharmacological and biological processes. The production of a therapeutic with a multitude of molecules represents a multi-targeted, systems biology approach to designing and manufacturing therapeutics. In the BRS “Intelligent Molecules” Expression System, multiple stem cell types are used to express SRM and thus the term “S2RM®” is used to denote that two or more types of stem cells are used to produce a synergistic effect. The combination of stem cells used to develop each therapeutic depend on the condition being treated, and the type of tissue underlying the condition. Included in the S2RM® is the BRS Exosome Delivery System that drives the active molecules through the layers of tissue into the target site, effectuating regeneration and repair in all damaged tissue regardless of the depth or remoteness of the target.

Unlike other so-called stem cell therapeutics, such as induced pluripotent stem cells (iPSCs) and parthenogenetic stem cells, which are artificially induced “stem cells,” and suffer from genetic and epigenetic programming errors, the adult stem cells in S2RM® Technology are genetically and epigenetically normal. Further, the production of some “stem cell” therapeutics does not allow the molecules to be completely processed in the cell and allow for proper post-translational modifications. That is, the S2RM® Technology allows the stem cells to fully process the molecules to the point of exosome production and release, including complete post-translational modification with resultant normal molecular folding characteristics and moiety construction. Thus, unlike iPSCs and parthenogenetic “stem cells” that don’t produce the complete set of the natural molecules of a stem cell and also fail to complete the full assembly of each of the molecules, the true adult stem cells in S2RM® Technology display correct genetic and epigenetic coding, with complete post-translational modifications to produce a natural and full complement of molecules, and with our processing technology the molecules develop complete post-translational modifications and hence full efficacy and a natural safety profile.

6. BRS: The Company and IP

BRS was founded in 2009 by Dr. Greg Maguire and Peter Friedman, and incorporated in the USA in 2010. In 1997 Dr. Maguire, who was professor at the University of California, San Diego, began injecting stem cells into the eye and brain for the regeneration of neural tissue. Those initial studies led to the discovery that stem cells release molecules into the damaged tissue to orchestrate a healing response. Other studies showed that multiple stem cell types, not just one type, home into the damaged tissue. These early studies, along with the work at BRS by Maguire and Friedman, led to the development of BRS’ patents that center on the S2RMTM Technology whereby the molecules, called stem cell released molecules (SRM), from multiple stem cell types, hence the designation S2RM, are used to formulate our therapeutic products.

Our IP strategy includes procurement of domestic and foreign patents in key markets for novel technologies relating to BRS’ core technology as well as such collateral technologies which are intended for product development and commercialization. While we disclose compositions and processes in our patents, not all processes are disclosed and remain as intellectual assets (trade secrets) within only a few individuals in the organization, namely Dr. Greg Maguire and Peter Friedman.

Currently, BRS has filed patent applications in the US relating to our core technology.

6. BRS solutions to the market needs

Dry eye is estimated to be a $1billion/year market in USA. Restasis has been the drug of choice at a cost of about $200/month to the consumer, and Restasis doesn’t work well (10% above placebo, FDA summary basis of approval) and stings the eyes. I-Ease eye drops by BRS is much more effective than Restasis, doesn’t sting, and costs $70/month, or less.

Cataract is debilitating, especially in our aging population. Current treatment is a costly and painful surgery that removes the lens and uses an artifical replacement lens. BRS’ L-Ease is an eye drop that prevents and reduces cataract formation allowing the patient to avoid surgery. We currently sell Lens-Ease for about $100/month. Results are usually seen in 2 to 4 months.

Our drug development programs for: 1. Dry eye, 2. Cataract were chosen to be undertaken first based on the following criteria:

1.  Cost and time to gain approval. Topical products with easy clinical endpoints to measure, and large, easily accessible patient populations to recruit for clinical trials.
2.  Large and growing markets. Each condition represents a large and growing patient population world-wide.
3.  Unserved or underserved market. Each condition lacks effective products for treatment of the particular condition.
4.  Chance for FDA “Fast track” status that will prioritize and hasten the approval time.
5.  Because the products are very efficacious, we expect a relatively low number of patients will need to be recruited and tested because a low variability in the clinical endpoint results. Therefore the trials should be relatively low costs given the fewer patients needed in the studies.
6.  Market [Total market with specific segments on USA, South America, Europe, Middle East, India and South East Asia] -Dry Eye – Estimated market in USA is approximately $1Billion annually. World-wide market is unknown, but underserved and growing.
-Cataract – The only treatment for cataract is currently lens replacement, a costly and invasive procedure that patients would like to avoid if a product were available on the market.
7.  Barriers to Entry [competition]
-Dry Eye – The product Restasis by Allergan is currently the product of choice. Restasis is not very effective: 10% better than placebo according to the FDA’s summary basis of approval. Restasis is also stingy and very expensive. Ophthalmologists tell me they don’t like to prescribe Restasis because of the expense and lack of efficacy, but that they currently have no better choice. With FDA approval/world-wide registration we expect quick acceptance by ophthalmologists and significant script activity.

-Cataract
An aging population means a greater incidence of cataract. No other product is available to prevent or treat cataract. Only cataract extraction through lens replacement is available as a treatment; an expensive and invasive procedure that is not well liked by patients. Our only entry to barrier will be that those ophthalmologists performing lens replacement surgery who will not embrace a product that lowers their surgery rate and therefore lowers their income.

8.  Product manufacturing
Products will need to be manufactured in cGMP facility starting with first-in-man studies. The I-Ease for dry eye, and the L-Ease for cataract will need to be manufactured as sterile eye drops. The active ingredients are manufactured by BRS, Inc. in the USA. All other excipients in the products are standard, easily-sourced, inexpensive ingredients commonly known to pharmaceutical manufacturers.

9.  Sales and Marketing strategy
License each product to large pharma company with world-wide reach and appropriate sales force.

10.  Funding requirements
We estimate a $1.5 million investment to bring each product through phase II. At phase III we would license the technology to a large pharma company for the studies and to bring the products to market.

References

Editorial. Failure To Launch. Nature Biotechnology 31, 1 (2013) doi:10.1038/nbt.2482
Rantamäki AH, Seppänen-Laakso T, Oresic M, Jauhiainen M, Holopainen JM (2011) Human Tear Fluid Lipidome: From Composition to Function. PLoS ONE 6(5): e19553. doi:10.1371/journal.pone.0019553
de Souza GA, Godoy LM, Mann M (2006) Identification of 491 proteins in the tear fluid proteome reveals a large number of proteases and protease inhibitors. Genome Biol 7(8): R72.
Srinivasan S, Thangavelu M, Zhang L, Green KB, Nichols KK (2012) iTRAQ quantitative proteomics in the analysis of tears in dry eye patients. Invest Ophthalmol Vis Sci. 2012 Jul 31;53(8):5052-9. doi: 10.1167/iovs.11-9022
Brian P. Danysh and Melinda K. Duncan (2009) The Lens Capsule. Exp Eye Res. 2009 February; 88(2): 151–164.
Mingyuan Zhou, Joshua Leiberman, Jing Xu, and Robert M. Lavker (2006) A Hierarchy of Proliferative Cells Exists in Mouse Lens Epithelium: Implications on Lens Maintenance. Invest Ophthalmol Vis Sci. 47(7): 2997–3003.
Maguire, G. et al (2013) Stem cell released molecules and exosomes in tissue engineering. Procedia Engineering.
Maguire, G. (2013) Stem cell therapy without the cells. Commun Integr Biol. 2013 Nov 1;6(6):e26631. doi: 10.4161/cib.26631.
Maguire, G. (2014) Systems biology approach to developing “systems therapeutics”. ACS Med Chem Lett. 2014 Mar 6;5(5):453-5. doi: 10.1021/ml500061.