Entrepreneurship Summary

Over the course of the past year, we have initiated the production of human protein S by recombinant DNA technology in the PROS project. PROS is a new treatment for protein S deficiencies that allows for the administration of human protein S and will help it become available in large amounts by recombinant DNA technology using fermentation in organisms (bacteria and insect cells). Compared with current treatments on the market, the applications of recombinant protein S have many distinct advantages.

We identified our key customer base and their needs and we deeply investigated the obstacles encountered by doctors and patients in existing treatment methods, so that they can benefit as much as possible from our therapy, PROS. We included government medical sponsor institutions, clinicians, and ordinary consumers in our analysis. We have also received feedback from medical professionals in well-known hospitals and healthcare facilities, as well as from ordinary consumer patients, and various different corporations, all of whom put forward high requirements for the strength, novelty, and price of our product.

For long-term sustainability, our treatment scenario will focus on protein S deficiencies, both inherited and acquired. We took an approach that is focused on treating the root causes of these disorders: a lack of protein S, as opposed to simply treating the symptoms. We conducted a competitor analysis and learned that, at present, most treatments for protein S deficiencies are limited by their cost, side-effects, and an individual’s ability to tolerate certain drugs and treatments due to other existing health conditions. Therefore, we chose to focus on developing a therapeutic that can be produced on a larger scale, has no side-effects, and can be used by any individual. Therefore, our product, PROS, will have a wider range of applications.

We analyzed our stakeholder behavior. In order to get more support and feedback from them, we wrote different versions of profiles, business plans and other content. We aimed to build stronger partnerships with our stakeholders, especially our suppliers and distributors, whose support will greatly optimize our production costs.

We developed a marketing Gantt chart for PROS technology transformation and business practices. This planning diagram includes research and development planning, patent application planning, clinical registration planning, medical device planning, enterprise cooperation and data platform planning and upscaling. It will take about four years to develop, promote and enter the market.

Our long-term impact is to bring new solutions to treat protein S deficiencies in individuals. We will influence the application and development of such a therapeutic. Our solution will become a more effective, safer, and convenient treatment option for disorders that include COVID-19, protein S deficiency, traumatic brain injury, and many others. This would not only upend existing treatments but also bring benefits to under-resourced areas. The benefits of our project, including its low-cost and safety, would allow more patients to afford it and would give local hospitals more opportunities to treat patients.

Please refer to the specifics about our product and proposed implementation at the link below.

For our project we also conducted a thorough SWOT analysis, which can be seen below, to assess its strengths and weaknesses.

1. Potential Customers and Unmet Needs

Doctors, patients and healthcare organizations can benefit from PROS in different ways. Not only does PROS optimize treatments, but it also reduces the risk of negative side effects in patients. PROS is a revolutionary treatment solution because it has the potential to treat a variety of serious disorders, is easily administered, and does not pose any significant health or environmental risks. We will analyze it from the perspectives of core competitiveness, user analysis, and demand satisfaction.

1.1 Inherited and Acquired Protein S Deficiency

Protein S deficiency is a genetic disorder that increases the risk of blood coagulation, and the formation of serious and fatal blood clots. It is caused by various different mutations in the PROS1 gene. These variations are inherited in an autosomal dominant manner. protein S deficiency can also be acquired; this means that an affected individual does not need to have a variation of the PROS1 gene. Acquired protein S deficiency can occur as a result of other underlying conditions such as liver disease, traumatic brain injury, nephrotic syndrome, certain infections, the use of oral contraceptives, vitamin K deficiency, surgery, chemotherapy treatment, and even COVID-19. These varied conditions can alter the levels of protein S in the body. These conditions are associated with significant morbidity and mortality, and have limited treatment options. They also require a large amount of medical costs and resources. Administration of protein S to treat these disorders can provide an effective therapeutic which lessens the severity of these disorders, and is significantly more accessible.

1.2 Related Disorders

COVID-19

Coronavirus disease (COVID-19) is an infectious disease caused by the SARS-CoV-2 virus. This disorder is widely characterized as a respiratory disease, however this is not necessarily accurate. COVID-19 patients experience a variety of symptoms including pneumonia, inflammation, micro-vasculature dysfunction, hyper-coagulation, nervous system damage and multi-organ failure. Moreover, there are multiple long-term complications, which are still not well-characterized.

Any of these symptoms can be responsible for COVID-19 related mortality. However, thrombosis (abnormal blood-clotting) is the leading cause of death in severe COVID-19 infections. Researchers have linked these abnormal blood-clotting events in COVID-19 with a decline in protein S levels (Faiez 2020).

There are multiple causes for the decline in protein S levels following a COVID-19 infection. In severe COVID-19 infections, there is an extreme immune response, known as thrombo-inflammation, which occurs by overproduction of cytokines, small molecules which regulate the activity of cells. COVID-19 can cause a “cytokine storm,” which damages cells, and leads to organ failure. It also causes a decline in protein S levels, causing many mini blood clots (microthrombi) throughout the body. Autopsies of patients with severe COVID-19 infections have revealed the presence of these clots in the lungs, causing extensive lung damage, which is a major cause of COVID-19 related deaths (Chatterjee et al. 2020).

A decline in protein S levels during COVID-19 can also be explained by the structural similarity between protein S and the spike protein found on the surface of the virus. This spike protein is recognized by the immune system. Following an infection, the immune system produces antibodies against the spike protein to neutralize the virus, but these antibodies can attack the structurally similar parts of protein S, leading to a decline in its levels (Pilli et al. 2016).

Overall, severe COVID-19 infections have been shown to cause a decline in protein S levels. However, research has also shown that in patients who already have genetic protein S deficiency prior to any infection, the effects are much worse. In a COVID-19 case report, preliminary protein S deficiency has been shown to cause ischemic stroke (Ali et al. 2021).

Essentially, it can be concluded that severe COVID-19 infections cause a significant decrease in protein S levels, and increase the incidence of abnormal or fatal blood clotting and inflammation. Researchers studying the direct correlation between protein S deficiency and COVID-19 suggest that administration of protein S in severe COVID-19 patients can be an effective therapy, and can be used as an alternative to traditional treatment. Administering protein S can also be used as a preventative therapeutic for patients with protein S deficiency in order to prevent the occurrence of stroke.

Pregnancy

Protein S levels also decline significantly during pregnancy, and protein S deficiency is associated with frequent miscarriages. According to the CDC, pregnant individuals are five times as likely as non-pregnant people to experience blood clots (CDC 2022). The major problem with blood clots is that they can travel to the lungs, causing a pulmonary embolism (PE), which can be fatal. Pulmonary embolisms are one of the major causes of pregnancy-related deaths in the U.S (CDC 2022).

We consulted Dr. Kimberly Herrera, a maternal-fetal medicine specialist at Stony Brook University Hospital. Dr. Herrera informed us that, because there is no available drug that replenishes protein S levels, mothers are often prescribed drugs such as Lovenox and Heparin in order to treat the symptoms of protein S deficiency.

Traumatic Brain Injury

Traumatic brain injury (TBI) is a neurodegenerative disease that affects 1.5 million people in the United States every year. Costs associated with TBI are estimated to be around $48.3 billion annually. Rehabilitation associated with TBI cost an average of $196,460 in the first year following injury. In patients that do not require rehab following a TBI, an average cost of $15,000 is spent in hospital and doctor bills along with clinical testing and medications. In those who experience severe TBIs, the lifetime costs can reach a staggering $4,000,000 (Swope, Rodante P.A.).

Following the initial tissue damage, secondary injuries that occur following a TBI include inflammatory responses, oxidative stress, interruption of blood flow to tissues, cell death, excitotoxicity. Cerebral edema leads to elevated intracranial pressure and decreased perfusion and limits oxygen supply to the brain which all contributes to cell death. In order to maintain fluid homeostasis, the brain will upregulate aquaporin-4 (AQP4), which is a water channel in the end processes of astrocytes that work to reabsorb the adema. The inflammatory response followed by TBI triggers the release of neutrophils, macrophages, leukocytes, cytokines, and collectively, these specialized cells increase oxidative stress. Increased oxidative stress activates microglia at the site of injury to repair the damage. The initial tissue damage that occurs is permanent, but secondary damage occurs at a slower rate, and, being given an appropriate treatment immediately following injury could improve prognosis of this neurodegenerative disease (Wang et al. 2020 ).

Currently, no effective treatment has been discovered for traumatic brain injury. One trial found that the administration of human plasma-derived protein S in mice 10-15 minutes after the onset of injury is able to reduce the extent of multiple secondary injuries in traumatic brain injury, and therefore improve prognosis. Motor function was measured in the protein S treated group and showed a 56% increase in the performance of the left front limb when compared to the control group which was treated with phosphate buffered saline (PBS). Differences between the control groups were greatest when protein S was administered at 24 hours following injury (Wang et al. 2020). Immunohistochemistry also revealed that the infiltration of CD11b+ leukocytes was significantly reduced around the site of injury in the group that was treated with protein S. Not only can treatment with protein S lead to substantial improvements of edema and fine motor coordination, but it also contributes to the mitigation of progressive tissue loss and suppresses apoptosis. Therefore, administration of human protein S can significantly improve the prognosis and life–expectancy of patients with TBI (Wang et al. 2020).

1.3 Current Protein S Deficiency Treatments

There is not a specific therapy for patients with protein S deficiency. The use of anticoagulant therapy, however, can be effective in the treatment and prevention of blood clots in patients with inherited protein S deficiency.

Anticoagulant therapy includes the use of drugs like heparin and warfarin that thin the blood and make it harder for the blood to clot. The choice of drug, specific dosage, and duration of anticoagulant therapy will vary among affected individuals. Factors influencing treatment decisions include the severity and frequency of blood clots, potential drug and dietary interactions, an individual’s personal preference, and age or overall health. Some individuals with a severe form of protein S deficiency may remain on this therapy for life. Special care must be taken if warfarin is used because of the risk of warfarin-induced skin necrosis.

Some individuals with protein S deficiency who have never had a blood clot will not need any treatment, except at times where there is an increased risk of blood clot formation such as during surgery, pregnancy, immobilization or trauma. Some individuals with a strong family history of developing blood clots may receive preventative therapy.

Genetic counseling may also be of benefit for affected individuals and their families.

Anticoagulants

Patients with protein S deficiency are commonly prescribed anticoagulants. Some likely prescriptions are heparin, warfarin, rivaroxaban, apixaban and dabigatran (Cleveland Clinic). Various pharmaceutical companies create and manufacture these drugs.

Heparin and Warfarin

Both drugs are commonly prescribed together. Prescribing Heparin prior to Warfarin will prevent widespread clotting (Cleveland Clinic). Because generic and brand-name Warfarin produce similar results, patients can opt to use generic Warfarin to save money (Dentali, et al., 2011). Despite its accessibility, Warfarin is associated with many adverse side effects such as, increasing bleeding and thrombotic events (Pirmohamed, 2006).

Rivaroxaban

Rivaroxaban (Xarelto) is an oral anticoagulant that is used for the treatment of deep vein thrombosis (DVT). Rivaroxaban can also be used for the prevention and blood clots in patients with atrial fibrillation. This anticoagulant can also be taken in conjunction with aspirin to lower the risk of heart attack and stroke in patients with a history of coronary artery disease. Rivaroxaban is available in a tablet and a powder for suspension. Unlike Warfarin, patients taking Rivaroxaban are not required to get a blood test to monitor prothrombin time. For DVT prevention following surgery, adult patients should take 10mg 1x/day for 12 days. For the prevention of DVT and PE, adult patients should take 10 mg 1x/day for at least 6 months in conjunction with a blood thinner. For treatment of DVT and PE, adult patients should take 15mg 2x/day for the first 3 weeks, followed by 20mg 1x/day as prescribed by the doctor (Rivaroxaban (oral route), 2022).

Lovenox

Lovenox is an anticoagulant that is used for prophylaxis of DVT and PE. Lovenox is used following certain surgeries as well as for patients who are bed ridden and at an increased risk of blood clots due to being immobilized. Chest pain or a heart attack may also be treated with Lovenox. This anticoagulant is injected subcutaneously or as an IV infusion. Lovenox is a single use, prefilled syringe and may be self administered by the patient. Medical tests are required while taking Lovenox to determine how long treatment is required (Sinha, 2022).

Eliquis (apixaban)

Eliquis is an Xa inhibitor anticoagulant that is used for the prevention of a stroke or VTE that is due to an irregular heartbeat not caused by heart valves. It is also used for the prevention of VTE in patients who have a history of blood clots, PE, and to prevent clot formation after a knee or a hip replacement. Eliquis is a tablet taken orally, 2x/day. Eliquis is very expensive as there is no generic available on the market yet. Another con of this medication is that in an emergency situation, it is more difficult to reverse the effects of Eliquis than it is to reverse the effects of Warfarin (Anderson, 2015).

1.4 Negative Side Effects of Current Treatments

As stated before, protein S deficiency is a genetic disorder that increases the risk of blood coagulation. There is no specific treatment for those with Protein S deficiency; treatments usually target symptoms associated with the disorder and severe untreated outcomes, like deep vein thrombosis (DVT) and pulmonary embolism (PE). DVT occurs when blood clots in veins far from the heart, like in the lower legs or arms. Blood clots initially formed by DVT can dislodge and cause PE, which is when a blood clot gets stuck in an artery of the lungs, blocking blood flow to part of the lung.

Patients with protein S deficiency are usually treated with anticoagulants in order to lower any present blood coagulation.

Warfarin

Warfarin is considered an essential drug by the World Health Organization, was the most popular anticoagulant in use before 2010 (Amareni) due to its ability to prevent and treat strokes, heart attacks, and DVT. The blood clotting process is complex and involves over 14 types of clotting factors. Warfarin is taken orally, and it denies access to Vitamin K, an essential vitamin, to multiple clotting factors near the entrance of the blood clotting pathway. This prevents those clotting factors from becoming biologically active, but it also gives Warfarin a slower onset time due to the time it takes to deny access to Vitamin K (Nature Video, 2018).

Warfarin interacts with many common herbs, over-the-counter medications, antibiotics, and vitamins. Some include garlic, St. John’s wort, Coenzyme Q10, aspirin, ointments, and skin creams that include aspirin, Pepto-Bismol®, Alka-Seltzer®, ibuprofen, and supplements with vitamin K. Warfarin can also interfere with antibiotics like metronidazole, nafcillin, and erythromycin; thyroid medicines like levothyroxine (Synthroid®), heart medication like Amiodarone, and seizure medication like Carbamazepine (University of Iowa Hospitals & Clinics, 2017).

Most side effects of Warfarin center around its ability to make a patient bleed more than normal. Mild side effects include heavy periods, nosebleeds, and gum bleeding, but more rare and serious side effects include blood in stool (poop) or urine, coughing up blood, bleeding from a cut or injury that won't stop, or strokes and brain bleeding (UK National Health Service, 2022).

Patients on warfarin have rates of minor bleeding at 15% and major bleeding at 0.4% to 7.2%. Major bleeding is categorized as such when the patient requires admission to the hospital, a fatal hemorrhage, bleeding at a critical site like the brain or retroperitoneum, or bleeding that requires transfusion of at least 2 units of packed red blood cells. Patients with major bleeding are several times more likely to die within a year of the occurrence. DVT patients on warfarin in particular are at a higher risk of bleeding when compared to patients on warfarin for atrial fibrillation (Amareni).

In the years since Warfarin's creation, research and development has included newer anticoagulants that are considered safer, such as Heparin, and a group of drugs called direct oral anticoagulants (DOAs), and Lovenox.

Heparin

Heparin is injected intravenously, and blocks the third clotting factor in the blood clotting pathway, in the "common clotting pathway" after both entrances. This prevents it from interfering with Vitamin K, and it interferes with less external chemicals than Warfarin because it interacts nearer to the end of the clotting pathway than the start (Nature Video, 2018). Heparin works more quickly than Warfarin because of its intravenous dose and direct interference with the clotting pathway instead of involving Vitamin K. It is also a larger molecule than Warfarin, which makes it unable to cross the placental barrier; therefore, it is safer to use during pregnancy than Warfarin.

Heparin's side effects are similar to Warfarin, including unusual bruising or bleeding, bloody vomit or vomit that includes clotted blood, chest pain, pressure, or squeezing discomfort, lightheadedness or fainting, sudden loss of balance or coordination, sudden trouble walking, sudden numbness or weakness of the face, arm or leg, especially on one side of the body, or sudden difficulty speaking or understanding speech in more serious cases (National Institutes of Health, 2017).

Lovenox

Taking Lovenox to treat protein S deficiency during pregnancy also has some risks. These risks include “excessive bleeding or bruising, vaginal bleeding, placental abruption—where the placenta pulls away from the inner wall of the uterus due to blood collection and or trauma” (Wisner, 2021). In addition, the company that manufactures Lovenox warns that pregnant people who have prosthetic heart valves should be cautious when using Lovenox, as it may result in valve thrombosis (Lovenox).

Rivaroxaban

Side effects of Rivaroxaban include, but are not limited to, back pain, bleeding gums, bowel or bladder dysfunction, coughing up blood, difficulty breathing or swallowing, dizziness, headaches, paralysis, prolonged bleeding from cuts, and vomiting of blood or material that resembles coffee grounds (Rivaroxaban (oral route), 2022).

1.5 Immense Medical Expenses

According to GoodRX, an online service that averages the uninsured price of medications, Warfarin generally costs $16 for 90 5mg tablets, Sodium Heparin generally costs $47 for 1ml of 5,000 units/ml, and Lovenox generally costs $95 for 12 40 mL syringes (GoodRX, 2022). For a genetic disease like Protein S deficiency to be treated effectively, anticoagulant medications would have to be taken for life.

Warfarin, taken orally and sold as a tablet, should be taken daily. Assuming a dose of 5mg per day at the price above, the average cost of Warfarin per year would be about $65 per year for an uninsured patient.

Heparin, taken intravenously, is usually used for surgical procedures or medical procedures of similar serious nature. According to Pfizer, the most widely used dosage is 5,000 units 2 hours before surgery and 5,000 units every 8 to 12 hours thereafter for 7 days or until the patient is stabilized (Pfizer, 2022). Averaging this to one dose before surgery and one dose every ten hours afterward for seven days, a sodium-Heparin solution would cost approximately $836.6 to administer to an uninsured patient during a serious medical procedure at a cost of $47 for 1ml of 5,000 units/ml.

Lovenox, administered as a subcutaneous injection either by a medical professional, or in dire situations, by the patient themselves, similar to an EpiPen's intramuscular injection. Lovenox can be used to prevent DVT in bedridden, seriously ill or pregnant patients after surgery, or can be administered during hemodialysis and heart attacks. It is also used to treat present DVT or DVT that advances into PE. In general, Lovenox is administered once every 8-12 hours for 8-10 days following major surgeries or other medical conditions (MedBroadcast, 2022). Assuming an uninsured patient is administered Lovenox once every 10 hours for 9 days at a price of $95 for 12 40 ml syringes based on the data above, they would have to pay $171 simply for the administration of blood thinners on top of their surgical costs.

Genetic counseling and testing is also highly recommended for patients with protein S deficiency, or a history of DVT and PE in their families. However, this presents a significant cost barrier. The cost of genetic testing can range anywhere from under $100 to more than $2,000 (MedlinePlus). For protein S deficiency, usually more than one test is necessary since there are a variety of mutations associated with the disorder, and sometimes, multiple family members must be tested to obtain a meaningful result, further increasing the cost. Companies such as PGxome typically charge up to $3000 specifically for genetic testing of the PROS1 gene for protein S deficiency (PGXome).

For people with protein S deficiency, or family history of related disorders, genetic counseling is also often recommended. For uninsured individuals, this can cost over $150 per hour per consultation (CostHelper).

All of these factors create significant cost barriers for the treatment of protein S deficiency.

In terms of disorders such as TBI, for which protein S can possibly serve as a treatment, there are also significant cost barriers, as mentioned before. Costs associated with TBI are estimated to be around $48.3 billion annually. Rehabilitation associated with TBI cost an average of $196,460 in the first year following injury. In patients that do not require rehab following a TBI, an average cost of $15,000 is spent in hospital and doctor bills along with clinical testing and medications. In those who experience severe TBI’s, the lifetime costs can reach a staggering $4,000,000 (Swope, Rodante P.A.).

1.6 Occupation of Medical Resources

Using Stony Brook University Hospital as an example, the medical/surgery department had 373 beds, and 24,880 surgeries per year in 2021, or an average of 66 surgeries per day–18.3% of the possible surgery beds were occupied on any given day that year. Protein S deficiency by itself does not cause bodily trauma, but it increases the risk of deep vein thrombosis, pulmonary embolism, and complications during pregnancy, surgery, and severe illnesses like COVID-19. It is difficult to estimate the number of patients in beds at any given time who have illnesses directly deriving from protein S deficiency; or the amount of space and time they take in hospital beds since the reasons for their stays will generally be different. It is safe to say, though, that the caliber of medical complication for protein S-deficient patients is significantly higher than that of patients without protein S deficiency. After major illnesses and surgeries, they will need more trained medical professionals to assist them, a steady supply of blood thinners being administered every few hours to every few days depending on the complications of their procedure, and general staff and nurses checking their condition frequently. It is also safe to state that protein S-deficient patients will take up hospital resources for longer amounts of time as their bodies are thought to recover more slowly after severe trauma (Wang et al. 2020).

1.7 Potential Customers

Customer Group	Examples	Value Proposition
Purchaser	Medicare and Medicaid (Public National Health Insurance)	Medical insurance companies take into account the affordability of products, their relevance to the population, the comprehensive interests of enterprises and the safety of the product. The cost-performance ratio between curative effect and product pricing determines whether insurance will cover the expenses of medical devices. Good pricing, safety, and applicability makes them more likely to cooperate and pay.
Purchaser	United Health, Kaiser Foundation, Anthem Inc, Centene Corporation, Humana, CVS, Health Care Service Corporation (HCSC), CIGNA, Molina Healthcare, Independence Health Group etc. (Private Health Insurance)	Medical insurance companies take into account the affordability of products, their relevance to the population, the comprehensive interests of enterprises and the safety of the product. The cost-performance ratio between curative effect and product pricing determines whether insurance will cover the expenses of medical devices. Good pricing, safety, and applicability makes them more likely to cooperate and pay.
Purchaser	Patients	Pricing is one of the most important considerations for patients, especially if they are uninsured. Ease of use, and accessibility are also important considerations, as well as therapy effectiveness.
Decision Maker	Doctor and Health Professionals	PROS allows for a wide range of uses, in patients with a variety of disorders that are associated with protein S deficiency. The novelty of this treatment as well as its far-reaching implications make it a good option for medical practitioners. For doctors, reccomening PROS as therapy is beneficial for the treatment of patients with various different disorders.
Users	Doctors	For doctors, PROS is attractive because of its effectiveness and therapeutic feasibility. Its biological safety is guaranteed by the safety of the materials used to produce it, and by future safety tests. It offers a better alternative to currently available treatments on the market.
Users	Patients	Price and ease of use is typically the focus for patients. PROS might not be cheaper than Heparin or Warfarin initially, but it is definitely a safer alternative that does present immense cost. Further industrialization and cost-analysis of the product will aid in fine-tuning it to make it more accessible and attractive. It is easy to use, and we have drafted a comprehensive user manual for doctors AND patients to refer to with product information.

First Potential Customers: Hospitals

In the future, PROS will be required to undergo clinical trials in order to determine safety and efficacy. Partnerships for this, as well as implementation would likely be in a hospital or healthcare facility. We have consulted with doctors at the Stony Brook University Hospital about the feasibility of our product. These interactions are described above, and in our Integrated Human Practices section. After in vivo testing and a variety of other steps such as the refinement and complete development of our product, clinical applications can commence. The use of such an injectable therapy such as PROS is not novel to doctors, so the implementation of the product in clinical trials and the healthcare setting is not particularly burdensome.

Second Potential Customers: Pharmaceutical Companies

Wayne Pharmaceuticals
We reached out to Wayne Pharmaceuticals, based out of Cordele, Georgia. Wayne Pharmaceuticals is one of the only Black-owned pharmaceutical companies in the United States operated by Dr. Wayne Whitest, M.D. Wayne Pharmaceuticals resides on a campus with the MD Whitest Medical Institute, which is located in a socio-economically disadvantaged neighborhood in Georgia. Wayne Pharmaceuticals and the MD Whitest Medical Institute focus on offering programs, products, and services to aid the surrounding community that experiences high morbidity and mortality rates. Dr. Whitest has developed his own line of healthcare products to help lessen the effects of heart disease and stroke which disproportionately affects African Americans in the United States. With African Americans spending billions of dollars each year on healthcare products including prescriptions and OTC medications, Wayne Pharmaceuticals seeks to provide high quality products that rival other pharmaceutical companies and divert funds back into Black communities to create jobs that will ultimately lead to economic growth. With Wayne Pharmaceuticals and the MD Whitest Medical Institute working to close the disparity gap seen in healthcare by the African-American community, this company could potentially help PROS Injectable Therapy reach a broader range of patients that typically see poor access to healthcare. The goal of our team is to work with companies that seek to raise public awareness of racial disparities seen in healthcare as well as improve the quality of care in underrepresented communities.

LucasPye Bio
LucasPye Bio is a large-scale biologics Contract Development Manufacturing Organization (CDMO), based out of Philadelphia, Pennsylvania that was founded by Lyles-Williams in 2018. This company offers various services including cell line development, bioprocess development, consulting and small scale bio processing. Lyles-Williams is an industry thought leader and she is also the first Queer, African-American woman to own and lead a biotech large-scale manufacturing company. 50% of their executive team are women and 85% are people of color. LucasPye Bio recently announced that they established their first contract manufacturing partnership with IndyGeneUS AI, which is an African-based genomics company. This partnership will work to develop targeted therapeutics that aimed to address health disparities.

1.8 Application Scenarios

When a patient undergoes testing and is found to have a protein S deficiency, the doctor can prescribe PROS to replenish the missing OR inactive protein S in the patient’s blood. Depending on the cause of the protein S deficiency, the disorder can affect a patient for either short or long term. There are different doses and maintenance doses depending on if a patient has a short or long term protein S deficiency. Patients can stop being prescribed the protein S injectable if their condition is one where there is short-term protein S deficiency, as soon as protein levels return to normal.

Note: a lot of the below information is restated from above, highlighting how PROS can be applied to a variety of disorders beyond simple protein S deficiency, and can improve the prognosis for multiple different conditions.

Inherited Protein S Deficiency

Protein S deficiency is a genetic disorder that increases the risk of blood coagulation, and the formation of serious and fatal blood clots. It is caused by various different mutations in the PROS1 gene. These variations are inherited in an autosomal dominant manner. Administration of protein S to treat this condition can provide an effective therapeutic which lessens the severity of the disease, and is significantly more accessible.

Pregnancy

Protein S levels also decline significantly during pregnancy, and protein S deficiency is associated with frequent miscarriages. According to the CDC, pregnant individuals are five times as likely as non-pregnant people to experience blood clots (CDC 2022). The major problem with blood clots is that they can travel to the lungs, causing a pulmonary embolism (PE), which can be fatal. Pulmonary embolisms are one of the major causes of pregnancy-related deaths in the U.S (CDC 2022).

Traumatic Brain Injury

Currently, no effective treatment has been discovered for traumatic brain injury (TBI). One trial found that the administration of human plasma-derived protein S in mice 10-15 minutes after the onset of injury is able to reduce the extent of multiple secondary injuries in traumatic brain injury, and therefore improve prognosis. Motor function was measured in the protein S treated group and showed a 56% increase in the performance of the left front limb when compared to the control group which was treated with phosphate buffered saline (PBS). Differences between the control groups were greatest when protein S was administered at 24 hours following injury (Wang et al. 2020). Immunohistochemistry also revealed that the infiltration of CD11b+ leukocytes was significantly reduced around the site of injury in the group that was treated with protein S. Not only can treatment with protein S lead to substantial improvements of edema and fine motor coordination, but it also contributes to the mitigation of progressive tissue loss and suppresses apoptosis. Therefore, administration of human protein S can significantly improve the prognosis and life-expectancy of patients with TBI (Wang et al. 2020).

COVID-19

Severe COVID-19 infections have been shown to cause a decline in protein S levels. However, research has also shown that in patients who already have genetic protein S deficiency prior to any infection, the effects are much worse. In a COVID-19 case report, preliminary protein S deficiency has been shown to cause ischemic stroke (Ali et al. 2021). Essentially, it can be concluded that severe COVID-19 infections cause a significant decrease in protein S levels, and increase the incidence of abnormal or fatal blood clotting and inflammation. Researchers studying the direct correlation between protein S deficiency and COVID-19 suggest that administration of protein S in severe COVID-19 patients can be an effective therapy, and can be used as an alternative to traditional treatment such as Paxlovid. Administering protein S can also be used as a preventative therapeutic for patients with protein S deficiency in order to prevent the occurrence of stroke.

1.9 Competitive Analysis

Core Competitiveness and Inventivity

There is no current direct solution to protein S deficiency on the market. Because protein S deficiency causes blood clots, current patients are prescribed blood thinners, such as Warfarin, to combat the symptoms of protein S deficiency. Such patients must live with the side effects of drug thinners for their entire lives. PROS, a protein S injectable, will offer a direct solution to this problem.

Competitor Profile

Competitive Frame

Unique Solution

PROS offers a unique, direct solution to a widespread deficiency. Similar to the prescription of recombinant human insulin developed in the 20th century, it achieves a safe, efficient, and cost-effective method of delivering protein S that is identical to natural human protein S, directly into the bloodstream. The dosage can be directly altered to match a patients need and pharmacokinetic profile, and does not present any adverse health effects. Neither does the development process include any potentially toxic materials or products, guaranteeing product safety. Future tests and clinical trials will be used to demonstrate this. Furthermore, it offers a novel therapeutic for disorders including severe COVID-19 and TBI. For COVID-19, there is no available therapy to reduce the risk of blood clotting associated with disease morbidity and mortality. For TBI, there is currently no direct treatment proven to improve prognosis. Our product proposes a novel therapy for both of these conditions that can significantly improve the prognosis of patients and reduce loss of life and function.

Efficiency

By injecting protein S directly into the bloodstream, our injectable therapy reduces the risk of other confounding factors, as well as the risk of side-effects that may arise. It also targets the root cause of related disorders such as PE and DVT by directly administering protein S. Because the protein is also injected intravenously, the product is fast and efficient. Dosages can also be altered, and protein S levels can be monitored in order to optimize the dose, as well as determine when a patient can stop using the product; for some patients, there will be long-term use, and for others, the product is only for the short-term.

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2. Design, Flexible Research, and Development

2.1 Safety of Materials

The long-term goal of our project is to produce a human recombinant protein S as an injectable medication through synthesizing it biologically in microorganisms–particularly E.coli. Our therapy proposes few risks to developers, patients, and health care workers, since we thoroughly analyzed risks behind manufacturing Protein S this way. Some microorganisms are pathogenic or harmful to the human body or surrounding environment, hence the necessary containment level and safety needs to be determined (NIH 2019).

E.Coli has a biocontainment level of BSL2. E.Coli plasmids aren’t capable of replicating in vertebrate cells, since they can only transfer the genetic material to each cell once. Even if laboratory workers are exposed to E.Coli plasmids directly, it should pose no risk, but as a safety measure, effective disinfectants should be left to set for 20 minutes, proper personal protective equipment and pipetting procedure should be used. E.Coli itself is a safe organism for laboratory use since it is unlikely to cause illness in humans, animals, or plants; therefore, it is safe for the environment even in the event of an accidental release. The genetic modifications in the PROS experiment are unlikely to mutate or cause further pathogenicity of the bacteria. Eukaryotic vector systems have been used extensively since 1983 for recombinant protein expression (iGEM Tec-Monterrey 2015), and they have been extensively tested for safety, confirming their neutral effect on human health (McWilliam 2006).

2.2 No Complex Technology and Fast Product Development/Iteration

We initially developed our product with the marketplace in mind, and with the goal of making our product as simple to develop and manufacture as possible. In this way, we are working on scaling up the production of our product to an industrial scale, and in our initial development, we used organisms and technology that are conducive for this. Additionally, we used mathematical modeling to further optimize the scale-up of our product. As a new product and part of a start-up company, PROS is also highly sensitive to the market. This makes it so that we can efficiently and effectively respond to market feedback and needs, and adequately meet demand for the improvement of our product. We can achieve this through fast and efficient research, and simple and not time-consuming or costly technological transformation and implementation. All of this will allow us to develop better products in short periods of time, in a way that matches market dynamics and fluctuations.

2.3 Diversified Team

We have assembled a multidisciplinary team of specialists who strive to produce a product that seeks to not only treat protein S deficiency, lessen COVID-19 symptoms, and reduce TBI injuries, but to also offer support to all patients seeking treatment. An important aspect of building and maintaining a diverse team is to recognize and celebrate cultural differences which will redefine a company's recruiting process, as we develop and expand our product. Diversity is critical for creativity, innovation, and professional growth, which will ultimately increase financial gains.

Having a diverse team will give PROS an advantage when we look to expand our product into new markets. Community connections, cultural responsiveness, and regional market knowledge will give our product a competitive edge, with a goal of reaching a more broad customer base.

2.4 Rich Academic Resources and Support

The academic support that we have received from Stony Brook University and a variety of individuals is one of our project’s greatest strengths, and has allowed our product to reach this stage of its development.

Institutional Direct Financial Support (totaling $41,200). Research Support Request (RSR), from the Stony Brook University Office of the Vice-President for Research, totaling $6,000, which was used to cover the cost of lab materials and reagents. There were also stipends that different iGEM team members received from different sources to support their research in the summer of 2022 came to a total of $41,200. The breakdown of support from different sources is as follows:

External Sponsor Commitments and Donations (totaling $25,755)

Target Customers

Target customers for the PROS medication include patients directly in need of the medication: those with protein S genetic deficiency, acquired protein S deficiency, those concerned about developing the acquired deficiency without preventative measures, and patients that have gone through severe bodily strain, such as a recent surgical procedure, recent bout of severe illness such as COVID-19, or recent severe injury, such as a traumatic brain injury. These should be considered short-term customers, as they will either be taking the PROS medication for a short time; only a few months to a year.

Long-term customers could include hospitals, pharmacies, and medical providers that purchase PROS therapy to keep in stock for their customers. The relative stagnancy and safety of their business models and their necessity to use PROS therapy, because it is more specific than any other medication on the market for protein S deficiency, renders them long-term customers. By a similar merit, a family that has a long history of genetic protein S deficiency could be considered a long-term patient since they would have to take the medication throughout their lives. Doctors supporting and buying the medication could also encourage more protein S research, development, and activity analysis.

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3. PROS Feasibility and Inventivity

3.1 Development Potential of PROS Treatment

The development potential of PROS treatment is mostly based on its continual presence as a genetic disease and its increased prevalence as an acquired deficiency as people age.

Protein S deficiency is caused by autosomal dominant mutations in the PROS1 gene. As mentioned above, acquired protein S deficiency can also occur as a result of liver disease, traumatic brain injury, nephrotic syndrome, certain infections, the use of oral contraceptives, vitamin K deficiency, surgery, chemotherapy treatment, and COVID-19 infection.

There will always be a certain number of people that require protein S deficiency treatment due to the constant nature of its heritability and of gene mutation. In addition, the conditions that cause acquired protein S deficiency, like chemotherapy treatment, exposure to COVID-19, and undergoing surgery generally increase in probability as someone ages. About surgery: according to a 2019 study, the average age of the English population in 1999 was 38.3 years and the average age in 2015 was 39.7 years, but over the same time period, the average age of people undergoing surgery increased from 47.5 years to 54.2 years old (Wiley 2019). According to the American Cancer Society, nearly two-thirds (64%) of survivors are 65 years of age or older, while only 1 in 10 are younger than 50 years of age (American Cancer Society). According to Science, the 2020 resurgence of COVID-19 in America was largely spread by those between the ages of 20-49.

According to The Administration for Community Living, a division of the U.S. Department of Health and Human Services, people aged 65+ represented 16% of the population in 2019. That is expected to be 21.6% by 2040. Generally, as people age, the need for protein S deficiency treatment will increase over time.

3.2 Proposed Implementation

There are several reasons for a patient to have protein S deficiency. A normal amount of protein S is categorized 60% to 150% inhibition (Mount Sinai, 2021). If a patient has a low amount of protein S, doctors should attempt to find the reason for the low protein S levels in the blood. There are several causes of protein S deficiency, some of which lead to a short term decrease of the protein, while others can be long term. For example, cases of severe COVID-19 can lead to a sudden decrease of protein S in the bloodstream for a short term. However, other causes of protein S deficiency, such as inheriting a mutation for protein S, may require lifelong treatment.

The normal ranges of protein S and protein C to be found in the blood were very similar, at 0.60-1.60 U/ml for protein S and 0.72-1.23 U/ml for protein C (Minuk, et al. 2010). Protein C and protein S also have similar functions, in which the two proteins bind together to break up blood clots. Because of their similar natures and amounts of both proteins that exist in the bloodstream, the proposed protein S injectable will be based off of the current protein C injectable, CEPROTIN, that is already FDA approved and used in hospitals.

Prescription Recommendations for PROS Injectable

	Initial Dose	Subsequent 3 doses	Maintenance dose
Acute Episodes, Short-term Prophylaxis	100-120 IU/kg	60-80 IU/kg Every 6 hours	45-60 IU/kg every 6 hours or 12 hours
Long-term Prophylaxis	N/A	N/A	N/A

These doses are the same as the dose recommended for use by CEPROTIN. Trials for the PROS injectable have not started, so it is unsure if these same dosages will be adequate for the PROS injectable. However, because protein S is very similar in terms of activity and quantity in the blood, if drug trials were to commence, initial trials may want to attempt dosage levels similar to the protein C injectable, CEPROTIN.

Regular monitoring of protein S activity level is encouraged to understand the impact of the injection, especially during the initial period in which PROS is administered. For those with long term protein S deficiency, continuous monitoring can be helpful to understand if the dosage of PROS is adequate to bring protein S activity levels back into a normal range. However, for those with short term protein S deficiency, monitoring of protein S activity levels will help physicians understand and adjust PROS doses. For example, if the patient is slowly starting to create protein S again, the patient can be slowly given diminishing doses of PROS or can stop receiving PROS treatment altogether.

3.3 Market Size

According to Fortune Business Insights, the global anticoagulant market size, in 2018, was an estimated USD 21.45 billion. It is projected to reach USD 45.50 billion by 2026. This insight is a reported compound annual growth rate of 9.9%.

According to a 2018 market analysis conducted by the the Journal of Thoracic Disease, the global market sales of anti-thrombotic (generally anticoagulant drugs) reached about 23.5 billion USD in 2013; about 2.7% of the global drug market. It has grown according to the expected expansion rate of reaching 25.9 billion USD in 2018 (Chaudhari, Hamad, & Syed., 2014). These trends have been predicted for a long stretch of time: a 2008 Nature article predicted the anticoagulant market reaching 6 billion USD in 2008 and 9 billion USD in 2014 (Melnikova, 2009). Although the market, generally dominated by warfarin sales, erodes over time as patents expire, Nature states that successful uptake of newer drugs like oral anticoagulants will drive future growth, to the point where the use of warfarin is “expected to become obsolete” (Chaudhari, Hamad, & Syed., 2014).

It can be concluded that the market for medication for protein S deficiency has grown significantly in the last two decades, leaving room for introduction and marketed development of new products like PROS treatment if it follows the trends for the market for general anticoagulant drugs–these drugs are currently used to treat Protein S deficiency in lieu of a more targeted solution.

3.4 Main Market Drivers

In today's society, drugs like Warfarin and Heparin are moderately effective treatments for blood coagulation, and, therefore, treatments for multiple cardiovascular diseases linked to blood clots. Without the presence of anticoagulants, there is an increased chance of blood clots to form, leading to multiple deadly diseases and heart attacks. However, the need for substantially more advanced and effective anticoagulant medications are the key drivers for the anticoagulation drug market. Displaying concern for the need to increase awareness of peoples’ cardiovascular health is another crucial driver for the anticoagulation drug market. For example, the introduction of the drug Eliquis by Pfizer and Bristol-Myers Squibb Company was wildly successful because there was concern over a lack of effective treatment. With the introduction of a substantially more effective treatment, the market will react positively in our favor. The unmet treatment needs is one of the major driving factors in this market.

3.5 Market Capacity

According to The Administration for Community Living, a division of the U.S. Department of Health and Human Services, people aged 65+ represented 16% of the population in 2019. That is expected to be 21.6% by 2040. Generally, the life expectancy for the U.S. population is increasing, although the COVID-19 pandemic has decreased the life expectancy in a manner opposite to predicted trends. A 2021 study found that life expectancy in the United States decreased from 78.9 years in 2019 to 76.6 years in 2021, but barring the effects of the global pandemic, life expectancy is generally rising.

With a higher life expectancy comes a higher aging population. As mentioned in the previous section on Market Size, the conditions that cause acquired protein S deficiency, like chemotherapy treatment, exposure to COVID-19, and undergoing surgery, generally increase in probability as someone ages. It can generally be assumed that with the increase of average life expectancy, the incidence of acquired protein S deficiency will grow over time. With this, the anticoagulant medication industry will be further developed by its increasing market demand.

3.6 Feasibility Analysis

Technical Feasibility

To determine how feasible it would be to produce PROS as a clinically approved medication on a large scale, the protocol, equipment and time-scale of the current laboratory experiment was taken into consideration. All portions of the experiment not relevant to the resulting protein being produced were not taken into account in order to streamline the process for efficient scheduling and development on a larger scale.

The experiment can be split into 7 important parts:

Our team was able to reach Step 4 in our 10-week experimental period. Preliminary steps necessary to complete the initial laboratory experiment and design its protocol took up a significant amount of time during the iGEM project period. The initial experiment brainstorming, proof-of-concept, collaboration with university faculty, and other iGEM teams worldwide is not factored into the schedule, but all were necessary to complete the wet laboratory portion of the experiment to some degree of success and create a viable protein with some degree of reliability. The Experiment and Results sections of this website provide more detail.

If we were to repeat this experiment with our current protocol and the knowledge we have after 10 weeks of testing and editing the protocol, we could theoretically reach Step 4 again within a month, assuming the Western Blot produced significant and testable results, then take one or two weeks to complete the purification of the product. From there, binding assays would be required to bind the protein and test its interaction with other molecules. We would hope for a similar result to that of protein C, which has already been studied, modeled, and characterized enough to use as a reference. After the binding assays, we would have to test the protein for its post-translational modifications, glycosylation rates, types of folding, and other factors.

To theoretically make a working drug on an industrial scale, we would need to continue perfecting expression of the protein so that it has all of its glycosylations and other post translational modifications as well as proper folding and proper removal of introns. The protein we have currently created has some significant differences to the human version. Our next step after the protein has been physically created and translational modifications analyzed, is to perform binding assays to protein C to see if the protein binds with it. Its place in the blood coagulation pathway in humans, as well as its general structure and functional similarity to protein C, means that it should bind. Physically making this happen will be difficult, but possible. The product can then be tested in mammals to consider short and long-term effects of using the protein product. Lastly, clinical trials and human studies would have to be conducted.

Then, the process to apply, explain the product and results of previous mammalian and human studies, and wait for FDA approval of the protein product, must be completed.

Safety

The main goal of the in-vitro experimental production of protein S was E. coli (prokaryotic) expression of the gene, which would be a similar goal when producing the drug on an industrial level. Due to the use of E.Coli for gene expression, production would incur some risk, although proper laboratory techniques and safety protocol, like wearing personal protective equipment, disposing of used materials into biohazard containers, sanitizing surfaces, and being conscious of one’s actions while working with the bacteria, could prevent serious risk. E.Coli infections are common, but most E.Coli bacterial strains are innocuous, and those that do cause infections are usually mild and easily treated. (Cleveland Clinic, 2019).

Economic Viability

Economic viability is measured by whether the benefits of a project outweigh its costs. Despite the high monetary and time cost of creating a PROS medication, the benefits to society at large would most likely outweigh the cost because PROS would be the first medication created specifically for protein S deficient patients that would be tested and refined for years in order to ensure its safety when taken lifelong.

The medical nature of this project brings about a certain necessity to complete it in order to provide assistance to patients that need it. This benefit of ensuring patient quality of life outweighs the cost of creating the medication, despite the thousands of dollars and years of deliberate, team-oriented effort that would go into its creation. In terms of monetary profit, the PROS medication has been shown to be profitable once it becomes more well-known, since it would be priced at a medium range compared to current alternative blood thinners like Warfarin, Heparin and Lovenox. This increase in profit over time renders PROS economically viable, although it would take years to become profitable due to the years of research and development required.

Expectation of Market Share

Our product’s primary focus is to act as a medical treatment for protein S deficiency. It could not be used as a general anticoagulant medication, since the administration of protein S to an individual that can already produce functional protein S would not make much of a difference. Our product development plan then, is entirely based on the possible medication, which should have a considerable demand because there is no specific alternative for patients with protein S deficiency besides taking more general anticoagulant medication.

The product should be implemented directly into the pharmaceutical and drug development industry, and does not have much implementation outside it.

Cost Input

The process of producing protein S as done in the initial iGEM project, and then theoretically scaled up to produce the drug on an industrial level would be limited by time, cost, and other factors in research and development. The largest limiting factor is time; cost is generally minute by comparison, but useful to analyze:

The cost of these important raw materials, including laboratory supplies and consumables, reagents for tissue culture, genomics, sequencing and other molecular services, would total to about $6,000.

Effective production facilities would need to include particular scientific equipment including consumables like pipette tips and non-consumable materials like a centrifuge; we estimate the cost of this to be $1,000.

Product Pricing

Since the materials for the product generally don’t cost much (the main cost of industrializing the medication would be the facilities and machinery), we could price our product reasonably compared to other intravenous alternatives used in surgeries. It is also important to note that protein S deficiency currently has no other specified treatment, which factors into the appropriate way to price it. Common anticoagulants like the pill form of Warfarin may be cheaper, at least initially,but this is for a generic, over-the-counter solution and does not target the same mechanism that the PROS medication would target.

As mentioned above, according to GoodRX, an online service that averages the uninsured price of medications, Warfarin generally costs $16 for 90 5mg tablets (GoodRX, 2022). PROS injectable medication would be priced at about $30 per dose. PROS’ high possibility of market growth makes it worthy of investment. Its relatively high price compared to Warfarin would be due to its “new” nature as a medication; costs would need to go into further research and development on its safety, and to the injectable mechanism by which to administer it, which Warfarin does not require by nature of being administered through a pill.

Operation Capacity

Our product’s primary focus is to act as a medical treatment for protein S deficiency. Therefore, our product development plan is entirely based on the possible medication and cannot generally expand into other industries. However, PROS treatment can carve out its own niche as a treatment specifically for protein S deficiency, which did not have more specific treatment alternatives beforehand.

Over the past two decades, the market for general anticoagulant medications has grown dramatically, with predictions to profit and grow further from the creation of new medications similar to oral anticoagulants, and theoretically similar to the PROS medication we intend to create. If all goes as planned, the product should be released after substantial quality control measures to ensure a high quality product at an affordable price point. Then, the product should be able to find its own market niche with no competitors in the industry at initial promotional stages. Theoretically, after a while of development and initial release, profits should be more substantial.

3.7 Scalability

Once refined and created on an industrial scale as a patented drug, the PROS treatment can be used to assist patients with genetic protein S Deficiency, but also acquired Protein S Deficiency. It can also be scaled to treat other health conditions, like concussions: It has been shown in mice with concussive traumatic brain injuries that injection of human Protein S can assist and speed up the recovery process (Wang, X. et. al, 2020).

3.8 Sustainability

Practicing sustainability is limited due to the nature of our project and the necessity of using sanitized, laboratory-safe materials with every step of lab procedures, but by being mindful of protocol steps, solutions and protein materials in use, we can practice laboratory sustainability by testing with small amounts of protein, fewer pipette tips and gloves, and less energy than necessary when going through protocol.

When the product is created on an industrial scale, 3D-modeling of the packaging and thoughtful consideration of materials to use can optimize sustainability while also creating, storing, and distributing the product as necessary for long periods of time.

3.9 Health Support for Underrepresented Groups

We have conducted extensive research on the lack of characterization of protein S in underrepresented groups. We focused on learning how protein S deficiency affects minority communities and learned of the need for more inclusive clinical trials to increase health support and resources for those from underrepresented communities.

Through meeting with stakeholders, interacting with African American and Indigenous communities, and based on the results of the evidence we collected, we learned many things that we aimed to integrate into the execution and implementation of our project.

In light of the evidence collected, we recognized:

In addressing the lack of proper diagnosis and treatment in African American and Indigenous communities, as well as the lack of knowledge of the effects of this deficiency on these groups we sought out a solution to combat both issues. Our research pointed us toward the problem of the race exclusivity of clinical trials as a whole in the United States. We created a guide that would help to further the development of more inclusive clinical trials based upon six principles:

In re-evaluating the way that clinical trials are designed in the United States, we will see the increased characterization of disease states in all racial and ethnic groups, which will hopefully aid in improving the care that individuals from underrepresented communities receive.

Addressing our third finding is not something that is easily fixed, and as a team we knew that we wanted to make a meaningful impact on our immediate community that would give students from underrepresented backgrounds opportunities not often afforded to them. We have been working with our P.I., Dr. Gergen on creating a research program that will be associated with our iGEM team that would allow students from a local community college, which is representative of minority and non-traditional students, to join a research team at Stony Brook University. Implementing a program like this would mean that students from underrepresented backgrounds are able to gain valuable research experience which will aim to increase minority representation in STEM careers with particular focus on medicine and research.

3.10 Patenting

After achieving successful expression of human protein S in E.coli, as PROS expands and commercializes, we plan to apply for a patent. We will begin with a provisional application which will detail our invention as well as its limits. While this patent will not include any specific claims, it will establish a “priority date”, which marks the first date that the patent was filed. Priority dates are critical in the case of another similar invention filing a patent in which the inventors associated with the earlier priority date will be upheld. After 12 months, we will file a “non-provisional” patent application which will incorporate our entire set of claims. Once the application is filed, details will be reviewed and an International Searching Authority (ISA) will research relevant documents that exist prior to our findings and record them into an International Search Report (ISR). Any objections made will be included in the ISR and we can adjust the patent as needed. We also plan to submit the Patent Cooperation Treaty (PCT) application, which grants us the option to pursue patents in 152 countries. If the ISA is unable to find any relevant documentation, then we could petition for an International Preliminary Examination, which evaluates the odds of being granted a patent. This could be a cost-effective solution for us because it would determine our chances before continuing to accrue the costs associated with obtaining a patent. After the examination process has concluded, the patent office will determine whether we are approved or denied. The entire procedure can take anywhere from a few months to years. We also plan to extend patent exclusivity which will motivate other companies to research and develop new therapies for protein S deficiency (Walker, 2022).

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4. Development Plans, Strategies, Funds, and Stakeholders

4.1 Stakeholders

Industrial Stakeholders

Alexander Knight, SVP Business Development for Cooley LLP. Cooley is a global law firm that was ranked a top law firm for US venture capital financings. Alexander Knight serves as senior vice president, leading Cooley’s global DEI-driven business development efforts. Passionately committed to seeing a more diverse entrepreneur and venture capital ecosystem, Alex plays a vital role in elevating the Cooley platform to serve the unique challenges and opportunities of underrepresented minority founders, investors and executives. He is a “go-to” adviser and connector to Black, Latinx and other underrepresented founders of high-growth companies – assisting them with targeted introductions to venture capital, executive talent and other high-impact resources that accelerate company growth. About half of their clients are in the life science sector, including healthcare tech companies, and because of this, we felt that having Alexander’s input would be beneficial as we navigate the target market for our injectable therapy, PROS.

After achieving successful results by expressing protein S in E.coli, we felt as a team that we needed to develop a business model that would not only deliver this therapy directly to the patients who are in need, but also consider the affordability and accessibility of PROS injectable therapy. Alexander, who has a background in working to increase the opportunities of BIPOC company founders and startups was an ideal expert in the field to communicate with. His first piece of advice was to classify the members of our team as specialists. For early stage founders it is critical to have a diverse team that specializes in different areas. He emphasized the importance of our story, and how that story will resonate with stakeholders. A large component of our project is highlighting where clinical research falls short in terms of the characterization of disease states in underrepresented communities.

On behalf of our team, Alexander reached out to the following companies:

Britt Gerald, Sr. Manager, Clinical Trial Diversity & Inclusion for Moderna. Moderna Inc. is an American pharmaceutical and biotech company based out of Cambridge, Massachusetts. Moderna develops mRNA therapeutics for use in vaccines and they conduct clinical stage trials using mRNA science to produce therapies for infectious, immuno-oncology, rare and autoimmune diseases. Currently, Moderna is using mRNA science to develop prophylactic vaccines for the flu, SARS-CoV-2, Respiratory Syncytial Virus RSV, human metapneumovirus and parainfluenza type 3 (hMPV/PIV3), cytomegalovirus CMV, Epstein Barr virus (EBV), HIV gp160, Nipah, germline HIV, and Zika VLP. Recently Moderna has reported positive data from clinical trials of their RSV vaccine and from their CMV vaccine. Moderna is also further increasing resources for their COVID-19 vaccine to create a wider pipeline.

Steve Worthy, Vice President, Business Development for Aetion. Aetion, based out of New York City, is a healthcare analytics company that presents data to manufacturers, pharmaceutical companies, purchachers, and medical treatment and technology regulatory companies. They utilized data science driven technology to help healthcare companies make strategic decisions that ultimately impact patients. Aetion developed the Aetion Evidence Platform (AEP), which is a software that quickly analyzes data to produce real-world evidence scaled to meet market needs.

Ingrid Green Jones, Esq. Sr. Corporate Counsel, Diversity Clinical Development for Pfizer. Pfizer is one of the world’s leading biopharmaceutical companies. They discover, develop and manufacture healthcare products, including vaccines and medicines. In 2021, their revenue was around $81.3 billion, with products sold in over 125 countries, 79,000 employees worldwide and 104 projects in their pipeline as of July, 2022, with 34 in phase 1, 34 in phase 2, 28 in phase 3 clinical trials, and 8 projects in registration. Pfizer has established The Pfizer Multicultural Health Equity Collective (MHEC), which works to better health equity of racial groups and underrepresented communities through advocacy and access, social determinants of health, diversity in clinical trials, and disease awareness.

Midstream Stakeholders

Midstream stakeholders include pharmaceutical distributors, medical and health institutions, and social welfare institutions. As we move toward clinical testing of PROS, we will focus on the Clinical Research Center (CRC) at the Stony Brook School of Medicine, where we will receive training to obtain a certification to conduct research on a human subject. Through Stony Brook University Hospital we will obtain an IRB approval, as well as file the Application for Approval to Conduct Research Activities at Stony Brook University Hospital. Research studies cannot begin until we receive both approvals (Guidelines & forms). Following a clinical trial, we will submit our results to the National Institutes of Health (NIH), as another midstream stakeholder to which we will register our product to ClinicalTrials.gov Protocol Registration and Results System (PRS).

Downstream Stakeholders

Downstream stakeholders include patients that could benefit from the administration of Protein S, like those with genetic Protein S deficiency, acquired Protein S deficiency, or those who have suffered from the effects of severe COVID-19 symptoms or traumatic brain injury. The development of drugs with fewer side effects has the potential to make a fundamental difference in the lives of patients. At the same time, from a public health perspective, the availability of alternative treatment options allows patients to choose more affordable drugs, thus allowing more patients to receive appropriate treatment. Used in combination with current available treatments, PROS will be a viable treatment option available to patients who are seeking to help manage the symptoms related to protein S deficiency. Along with the customer, downstream stakeholders also include individuals or organizations that provide support to a product. We will implement a team of nurses, pharmacists, and clinicians that will be available by phone to answer any questions that a patient may have in regards to the administration of PROS injectable therapy.

4.2 Risk Assessment and Response

PROS will adopt the standard operating procedure for quality risk management (QRM) in pharmaceuticals laid out by Pharmaguideline. QRM is used to assess, regulate, communicate with the team, and examine the risks that are associated with a pharmaceutical product. There are two fundamental principles of QRM:

Identifying hazards, examining, and evaluating the risks that are associated with exposure to those dangers can be managed by classifying the risks both qualitatively and quantitatively. We will qualitatively address a risk by categorizing it as ‘High’, ‘Medium’, and ‘Low’. Quantitatively measuring a risk will require using a number value of ‘1’, ‘2’, ‘3’, ‘4’, and ‘5’. The following table to determine the severity of risk management is adapted from Ankur Choudhary at Pharmaguideline.

After identifying the potential risk, plans for mitigation will be discussed by the team to determine the suitable actions that are required to minimize the outcome. As production of our product expands, we will establish a multidisciplinary team where we will designate experts in the field of production, engineering, marketing, and consumer relations to oversee QRM for PROS Injectable Therapy (Choudhary, A.).

4.3 Development Plans and Financial Projections

Development plans have been outlined throughout this document. We have organized our team of specialists, outlined our products and services, conducted a market analysis, identified potential competitors, and developed a strategy and implementation plan for PROS injectable therapy. A very important component of the development of our business is our financial plan and projections for our product. It is critical that the following statements are incorporated into our financial projections:

Because PROS is a new therapy, we can analyze the projected growth for the biologics market as a whole. According to the Biologics Market Report 2022-2032, the market is forecasted to create a profit margin of around 40-45%. There are some questions to consider as we enter the development stage of our product, including identifying if there is a need for product commercialization in order to upscale the biologics market; how is the biologics market evolving in the next decade and what would be the best investment option for the PROS product (Biologics market report 2022-2032, 2022). We foresee the direction of PROS moving to offer a more cost effective product that can be afforded by all patients. The biologics sector of the pharmaceutical industry has the potential to address a multitude of conditions and investing in research and development that cuts costs without sacrificing quality to ultimately offer the patient an affordable medication is how we hope to see the market evolve.

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5. Long Term Impacts and Beneficial Influence of PROS

5.1 Operational Planning

Current Business State

Currently, PROS is focusing on the research and development phase of the protein. At this point, the protein has been synthesized; however, purification would be necessary to further characterize and analyze the activity of Protein S. This is due to the fact that the protein was synthesized in E. coli, which may mean that the protein which was synthesized may have different functionality. We would like to conduct assays to see how it binds to Activated Protein C (APC), which is the mechanism that prevents clotting by degrading clotting factors. In addition to these binding essays, the protein would need to be analyzed for post-translational modifications and other factors that would influence improper folding, which would take years of laboratory testing. After testing functionality, the next step is to construct a model for injectable use. This model could be based on the model for Ceprotin, which is a protein C intravenous injection. Once the injection is modeled and the final product is made, the team will work with the FDA to get it approved. Then, we will work with a manufacturer to mass produce PROS. Additionally, we will conduct marketing to release the product to the public as an alternative to blood thinners. This product would be attractive as other current therapies do not provide a direct protein S administration. Instead, they work to curb the effects of the deficiency by treating blood clots. PROS would supplement the body’s protein S and can help target certain types of the deficiency, such as Type I, Type II, and Type III.

Growth Strategy

The research and development that was used to create the PROS medication, made public to an extent through iGEM, can help other iGEM teams conduct similar experiments with Proteins S and C. Literature searches, general research our iGEM team conducted on Protein S, and the journal we published including information on Protein S deficiency can allow PROS to broaden its customer base by spreading awareness of Protein S deficiency, which is generally not well-known. PROS medication can be used to treat those with genetic and acquired Protein S deficiency, and can theoretically be expanded to treatment of comatose and severely injured patients.

Ultimate Goal

PROS hopes to be a fully developed injectable drug that can be used by anyone who has been diagnosed with a protein S deficiency, with various dosages depending on patient condition. In the future, the injectable will be manufactured by a major company after having gone through multiple rounds of testing. Then, this therapy can be administered to patients by a physician. Even farther in the future, we aim to propose an even better injectable design that can be implemented for safe use at homes, to help patients with long term protein S deficiency. PROS will have a similar trajectory for at-home use when compared to insulin. The initial dose of PROS may be higher than maintenance doses. For this reason, and so that the doctor can manage the reaction and wellbeing of the patient when the drug is initially administered, the initial dose may be administered in an intravenous way in medical locations. This is what the current PROS treatment plan entails. However, once a patient has been monitored for the initial rounds of PROS treatment and has no adverse side effects, we hope to design a therapy so that patients may be free to administer PROS safely at home. This is the ultimate goal of our project.

5.2 Beneficial Influences of PROS

PROS treatment is primarily intended for those with genetic or acquired protein S deficiency. The functioning protein S within the PROS treatment would replace the non-functioning protein S in their bodies. This would allow their blood to clot at a normal rate and put them at a lower risk of deep vein thrombosis, pulmonary embolism, and serious medical complications after surgery or prolonged illness like COVID-19. The protein S in the injectable PROS medication also has beneficial influences not directly related to protein S deficiency: for example, the injection of protein S into mice suffering concussive traumatic brain injuries was shown to help these mice recover from injury quickly and more effectively (Wang, X. et. al, 2020).

5.3 Possible Adverse Effects of PROS

Needle injection within the bloodstream may give rise to adverse effects like inflammation of the subcutaneous layer of the skin and constrict the blood vessels (Usach et al., 2019). This prevents mobilization of Protein S within the bloodstream, thus reducing chances of it reaching the unwanted blood clot. In addition to the inflammation, many of these injectables aren’t administered by health professionals, but by patients themselves (NORD, 2019). Due to misadministration of injectables, it may increase patient susceptibility to external or opportunistic infection. Within the bloodstream, the foreign Protein S may be attacked by the patient’s natural immune system labeling it as a foreign substance. This will reduce the amount of the desired protein within the body that could bond and dissolve unwanted blood clots. Protein S administration has very low amounts of literature prior to our research, so it is difficult to know what allergic reactions could arise in patients through an injectable therapy (NORD, 2019). Allergic reactions may consist of swelling, dizziness, fever, hives, or rashes (American Academy of Allergy, Asthma and Immunology, 2020). Looking at the manufacture and accessibility of this drug on the market, there are a few caveats that could hinder the sustainability of this admisinstation. Nonrenewable injectables would increase plastic consumption, and increase the chances of it being misplaced or misused after injection.

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