Team:Aalto-Helsinki/Entrepreneurship

Aalto-Helsinki

JUDGING

Entrepreneurship

Stage 1

Eureka Moment

Finland is known as the sauna nation and we Finns are very proud of our sauna culture. In fact, if you visit Finland, a must do is to take off all your clothes and go to a little room heated to almost 100 ℃, where you sit naked with others and sweat. Since sweating in sauna is a huge deal, from the beginning of our iGEM project, we had some idea for sweat based analysis. During our literature search to understand different substances present in sweat, we found that many proteins are also secreted in sweat as part of our innate immune system. That is when we serendipitously came across an interesting peptide called dermcidin [1] . Dermcidin is known to have antimicrobial activity against a range of different microbes. Though initially we had many possible application ideas and different problems to solve, we realized that the peptide in our hands may be best-suited for solving the problem of acne. We realized that this was an every household problem, affecting millions in all age groups across different countries. To develop our product for acne, we first had to validate the market demand and product potential.

Focus Group Survey

Before developing a product, it was important for us to understand the consumers and their needs. So we started off with a consumer survey involving 194 people from different age groups between 16-60-year-olds. Our primary questions were directed towards understanding the depth of acne as a problem in consumers' day to day lives. Below is the list of questions and data we received.

About acne presence and severity among people surveyed:

Correlation between acne severity and mental wellbeing:

Product Potential and Customer Group Identification

After the extensive focus group survey to identify the depth of acne problem, it was quite evident that acne was indeed a serious problem among a range of different age groups. At least 60% of our responders have been diagnosed with acne at some point of their lives and at least 45% of our responders have suffered from acne for more than 1 year. Hence, we realized a need for a “continuous” treatment for acne. Further, to understand the relationship between acne severity and mental wellbeing, we carried out a correlation analysis. There was a clear effect on mental state among the people who have been diagnosed with acne, regardless of its severity. We did not, however, ask what was bothering them and to what extent acne affected their well-being. Similarly, acne had negatively affected the social interaction of our respondents. We realized that Acne vulgaris affects about 85% of young adults aged 12–25 years and hence forms our primary customer group.

The most commonly used products for acne among our focus groups were oral antibiotics, birth control pills, retinoids (Roaccutan), Ayurvedic medicine, finacea 15% gel (azelaic acid), over-the-counter topical medicines (e.g., benzyl peroxide, salicylic acid), isotretinoin Actavis, prescription topical medicines (antibiotic), laser or light treatment and natural treatments.

Of the 194 participants, 95 of them didn't report any side effects from the acne treatments. However, remaining 99 reported some kind of side effects while using the acne product or undergoing treatments.

Market Research

After identification of focus groups and realising product potential, we went for a careful market research to identify how much current acne treatments cost and what are the current acne products in the market.

Current acne treatment costs

The following cost ranges are based on approximated acne treatment expenses in the United States.

For over-the-counter daily regimens which includes a cleanser, a toner and a non-prescription medicated lotion, acne treatment costs approximately $30-60 (about 25-50€) or more per month. An acne dermatologist visit usually costs$100-200 (85-170€). For prescription antibiotics, topical creams and ointments - or for hormone therapy, from birth control pills to a more aggressive regimen - acne treatment typically costs $45-200 (40-170€) per month. In some contries, acne treatments, such as prescription drugs and topical creams, can be covered by health insurance plans. The treatment often requires 2-3 months to see improvement. Once acne clears, it must be continued to treat the skin to prevent breakouts. Additionally, for physical (cosmetic) treatments such as aestheticians facials or chemical peels, acne treatment typically costs$75-200 (65-170€) per session, with one or more sessions required per month. Once acne is cleared up, some patients choose to undergo additional treatments for scarring. These can include microdermabrasion and dermabrasion, which can cost up to $2,000 (1700€) or more, depending on the number of sessions needed. Light and laser therapies can be used for the treatment of acne. There is, however, insufficient evidence to recommend the routine use of these therapies. The treatment costs can range from$100-400 (85-340€) per treatment, and \$600-1500 (500-1300€) in total. Most patients will be with somewhere between 4-8 treatments (3 weeks to 2 months).

Current acne products

TreatmentAgent / DrugDisease severity- treatment availability/recommendation
MildModerateSevere
Tazarotene (Tazorac)
Tretinoin (Retin-A)
Topical antibioticsClindamycin (Cleocin T)
Erythromycin (Benzamycin)
Oral antibioticsDoxycycline
Erythromycin
Minocycline (Minocin)
Tetracycline
Trimethoprim / sulfamethoxazole
(Bactrim, Septra)
Vibramycin
(Oral) isotretinoinAccutane
Oral antiandrogenBirth Control Pills (e.g., Ortho Tri-Cyclen)
Spironolactone
OTC treatmentBenzoyl peroxide (topical)Benzac AC
Salicylic acid (topical)Salicylic acid
Azelaic acid (topical)Azelex
Topical DapsoneAczone

These medications have different mechanisms of action and properties: antibacterial, sebostatic (reducing sebum production), comedolytic (reducing comedone formation) and anti-inflammatory.

TreatmentAgent / DrugProperties
AntibacterialSebostaticComedolyticAnti-inflammatory
Tazarotene (Tazorac)
Tretinoin (Retin-A)
Topical antibioticsClindamycin (Cleocin T)?
Erythromycin (Benzamycin)
Oral antibioticsDoxycycline?
Erythromycin
Minocycline (Minocin)
Tetracycline
Trimethoprim / sulfamethoxazole (Bactrim, Septra)
Vibramycin
(Oral) isotretinoinAccutane
Oral antiandrogenBirth Control Pills (e.g., Ortho Tri-Cyclen)
Spironolactone
OTC treatmentBenzoyl peroxide (topical)Benzac AC?
Salicylic acid (topical)Salicylic acid
Azelaic acid (topical)Azelex
Topical DapsoneAczone

Acne medications are rarely reimbursed in general. Their availability as well as prices vary across different countries.

We had successfully assessed the market and the consumer problem. Now it was time for the product development.

Stage 2

Product Development Phase

Consumer preference and needs

We asked the 188 consumers about the product preference they had in terms of how they wanted to use the product. This was crucial to take into account our product design. 42% of them preferred it in the form of a cream, while 27% of them preferred it as a face mask.

Product Development Strategy

Our product PORIFI would be the first product in the market to use an antimicrobial peptide for acne treatment. We decided to incorporate the peptide in a hydrogel, which could provide a good environment for it.

Process Development Strategy

Producing an antimicrobial peptide using chemical synthesis on a mass scale is really expensive. A few grams of the chemically synthesized antimicrobial peptide costs around 1000 €.

For comparison, check custom peptide prices from Abm.
4 mg of Dermcidin ~1400 dollars (purified)
1-4 mg ~420 dollars (10 dollars crude)
0,1 mg we bought for 300 e

It is tough to beat the current products in the market this way. Hence, we decided to go for production of our peptide DCD-1L in E. coli as an economical alternative.

Another challenge was to produce an antimicrobial peptide in E. coli without killing the bacteria itself. So we designed a system of production with Smt3 fusion tags for the production.

Consumer Acceptability

Having designed the product and the process, it was important for us to see if the product was accepted by the consumers. The results of product acceptability can be seen below.

It was interesting for us to see that around 78 % people were ready to use a product from genetically modified bacteria. Further, if the product was identical to something naturally found in the body, 82 % people were ready to use it. This was a positive sign for us to proceed ahead in the development of product in terms of formulation and prototype development, packaging and quality testing.

Patent Screening

During the process of defining our project, we had briefly checked the patent to ensure that the idea we would work on has also legal foundation. However, we had been also told not to be discouraged if we find the patents of similar content and to carry on our scientific research if we feel that a problem is not explored enough.

After the meeting with the doctors from the "Biodesign Finland" entrepreneurial program and their advice, we decided to screen the patent applications and granted patents again. We used the online patent search tools, such as The Lens, Google Patents and European Patent Register, to find relevant patent information. We searched through abstracts and claims for the following keyphrases: dermcidin, dermicidin, DCD in combination with skin and acne terms. We also got support from the law students of Tec-Chihuahua iGEM team, as part of our collaboration.

As our project is related to biotechnology, we were interested mainly in the legal protection of biotechnological inventions. First, we familiarized ourselves with the basics of governing laws and procedures related to patent applications for different regions. However, we all study in Finland for longer term, so we looked closer into the procedures in European Union, where the rules on the scope and limitations of patent protection for such inventions are governed by Directive 98/44/EC. This regulation defines biotechnological inventions as products consisting of, or containing, biological material, or processes by means of which biological material is produced, processed or used. To patent such inventions, the general requirements for patentability must be fulfilled, i.e. novelty, inventiveness and industrial applicability.

In general, an European patent shall be valid for 20 years from the date of filing the application. However, renewal fees must be paid to the European Offices in respect of pending applications and later to the national offices in every contracting state in which a granted patent is brought into effect. More information can be found from the European Patent Convention.

Although first patents related to dermcidin were filed not earlier than about 15 years ago, some of the granted patents expired due to failure to pay maintenance fee. A few of granted patents are still in force in individual countries and several new applications are still pending. We listed the patents relevant to our project and described their legal status and most important claims in the separate document.

The document with the current patent situation can be found here.

Stage 3

Creation of a Product Using Synthetic Biology

Trust over Technology

Using our production strategy, we were able to scale up from a 5 ml batch to a 500 ml batch. For details on the design process for production and success with the small scale and large-scale production, please check out our Laboratory pages.

Prototype Development and Formulation

Prototyping comes into play when the design of the product needs to be verified during product development, because it demonstrates or proves the design. Our main motives for a prototype design were to:

a. Show the new product to investors.
b. Test an idea to see if it really works.
c. Test the design to see if it passes certain requirements.
d. Evaluate necessary improvements.
e. Get customer feedback.

There are 2 categories of prototypes:
1. "Looks Like" - Prototypes that look and feel complete but do not essentially function.
2. "Works Like" - Prototypes that function properly but do not look anything similar to the final product.

Our prototype falls under the first category. Contact us to check out our product prototype at the Jamboree.

Development of Packaging

For an interesting in-depth step by step design process of our product prototype, development of packaging and formulation please check the Applied Design page.

It is important to have a unique and an attractive package for a cosmetic product to be considered by the consumers. The packaging creates the first impression. Here is our design of the eco-friendly wooden box for product packaging. For mass production, a wooden package is most probably too expensive, but for the project we wanted to focus on a beautifully designed piece as well as a concept for a package that can be used after the product has ran out. In the future the package could be a polylactide (PLA) tube or a coated paper tube which would be easy to recycle or burn.

Quality Testing

We carried out quality testing experiments including a shelf-life study and the stability of our product. The Urlinski company, which designs and manufactures the equipment for production of cosmetics and medicines, pointed out that the biggest problem with preservative-free biomaterial based products is contamination. In our experiments we noticed, that some of our samples got contaminated in room temperature. We concluded that biobased products get too readily contaminated, especially if the product is kept in room temperature and handled with bare hands. Thus we tried out different formulations for our product with an approved cosmetic preservative: benzyl alcohol (3%) and carried out a shelf-life study. Another advantage of including benzyl alcohol into formulation was the fragrance it added to the product.

Stage 4

Production Scale-up Estimation

From the 500 ml batch, we were able to produce around 30 mg of the DCD-1L peptide. Our next step was to estimate the production cost for a semi-medium scale up process.

 *Semi- Medium Scale Up Process Production Cost Sheet Euros Stage 1 Raw material Organism cost 5 Culture medium 180 Inoculant cultivation (in terms of time) 60.3 Skilled Labor cost involved 160 Stage 2 0 Fermentor usage - SCIGENICS BIOFERM - IM 42 Energy involved for the production process 60 Skilled Labor cost involved 100 Miscellaneous (time + other Raw material used) 210.6 Stage 3 0 Purification 0 Working Cost (Raw materials and Facility utilisation) 250 Process 800 Energy utilised 80 Skilled Labor cost involved 320 Miscellaneous involved during conversion process; 140.4 Additional Energy cost 100 Stage 4 0 Digestion and final purification 2010 Total 4518.3

*Costs Inclusive of Time, space, Labor skill expertise, energy and instrumentation

# The above mentioned prices are calculated values and quoted to calculate approximate production cost. Subject to vary during real time conversion / Scaleup process.

100 mg of DCD-1L is recovered from per kg of input, resulting in 4518 euros. The approximate range, however, will be 3500-4500 euros for 100 mg of DCD-1L. Hence we have brought down the cost of DCD-1L production 10 times lower using synthetic biology technique as compared to chemical synthesis. Costs will go down with larger batch sizes and a continuous fermentation process.

Our cost of the product needs to be estimated yet based on final packaging and formulations used. Below, there is a rough estimation of the monthly product costs for the customer. Note! The estimation below does not take into account possible preservatives and exfoliating agents, which will also be added in the final product.

Price calculation for PORIFI
Ingredient€/package% of total priceIngredient cost
dH2O price0.130.10 %1.1€/L
CMC price0.020.01 %5€/kg
DCD price133.299.89 %37€/mg
Peach seeds, Benzoyl alcoholTBDTBDTBD
TOTAL PRICE133.35
Product Usage
Package volume120ml
DCD weight0.03mg/1ml
DCD weight4mg/package
Product per use3ml per application
no. of applications40
Application frequency2times per week
A package lasts for5months
Porifi treatment cost29.34€/month

Funding

The most crucial part of our product development and entrepreneurship journey is funding. We have so far collected around 80,000 euros for the initial RnD and product design. But we would be applying for an initial seed funding round to carry out further activities in research and development and product improvement. We continuously seek to improve the product based on interviews and consultation with people from the industry. We have currently pitched our product idea in front of experts from the Pharma and Bio Industries. We are yet to have investors pitch round which would be done only after the seed funding rounds are over.

Regulatory Approval and Safety Testing

A big part of working with genetically modified organisms is safety and this is why there are several laws and regulations concerning them. If our product is considered to be a GMO, which without the cellulose binding module is not exactly certain since despite the production method, the end product is basically identical to the natural dermcidin, we also have to consider these as part of the launching plan. It is to be noted that since our product would most likely fall under cosmetics, where the regulations are less strict than with the medical or agricultural fields. The time to get the actual product to the market should also be significantly shorter. To confirm these assumptions, we would need significantly more legal expertise and would therefore consult a very specialized lawyer before applying for permission to use our dermcidin-hydrogel as a cosmetic product.

In general, there are international treaties on safety in biotechnology like NIH Guidelines, The Codex Alimentarius, OECD's Recombinant DNA Safety Considerations, also known as the Blue Book, The Cartagena Protocol of Biosafety and WHO's Biosafety Programme. Based on these, there are several EU directives and regulations, which have also been implemented into Finnish national legislation in the form of e.g. The Gene Technology Act (Geenitekniikkalaki 377/1995) and lower level national regulations [2].

If we have understood correctly and there hasn't been any recent changes, placing on the market of non-food and nonfeed GMO-products is done according to DIRECTIVE 2001/18/EC in Finland. First, the notifier contacts the competent authority in Finland who contacts the European Commission. Then, if the other member states won't object, the application proceeds to the European Committee and can be approved with a qualified majority or move forward to the European Council and Parliament, but the execution decision will be made by the Committee. GTLK (Geenitekniikan lautakunta) is the Finnish national competent authority in issues related to the Act of Gene Technology and it coordinates the functions of the supervisory authorities.

Detailed Plan for Product Development, Company Establishment and Launch of Product

Here is our detailed timeline in weeks and strategy for product development and company establishment. It can be also accessed as an Excel file.

References

[1] Schittek, B., Hipfel, R., Sauer, B., Bauer, J., Kalbacher, H., Stevanovic, S., ... & Rassner, G. (2001). Dermcidin: a novel human antibiotic peptide secreted by sweat glands. Nature immunology, 2(12), 1133-1137.