Evaluation of Squalestatin 1 as an Enzyme Inhibitor for Lowering Cholesterol Presented by Sam Noor-Mohammadi Daniel Feze April 28, 2008 Advanced Process Design

Reversal of Medication Labels

Agenda Problem Statement Background Drug Model Production Economics FDA Manufacturing Economics Demand Model-Price Model Conclusions

Problem Statement This work evaluates the economic potential of a new enzyme inhibitor for lowering cholesterol levels in human plasma. We looked into FDA Process Manufacturing and Production Economics and Pricing of the Drug

Cholesterol Produced by liver cells and acquired through diet and natural production Three major types: High density lipoprotein (HDL) “good cholesterol” Low density lipoprotein (LDL) “bad cholesterol” Triglycerides Important in transfer of blood constituents (ex. lipids) Important in cell survival

Formation of Plaques Formation of Plaques in the Arteries LDL are retained by artery walls while moving the cholesterol from liver to peripheral tissue. This accumulation is responsible for the hardening of the arteries

Health Facts High Cholesterol Leads to Heart Disease 29% of Heart Disease Related Deaths in the U.S Each Year 17.5% of People Over 20 Years of Age are at Risk for Heart Disease and Stroke

Health Facts Good Serum Cholesterol Level is 199 mg/mL In 2005-2006, 16% of American Adults had Levels Over 240 mg/mL For Women Over 60 Years this Level was Higher than Men For Women and Men 20-59 Years the Same Levels Were Observed

Solutions to Lowering Cholesterol Low Fat Food, Exercise Herbal and Nutritional Solutions Statin Drugs Vitoryn Zocor Zetia Lipitor Pravachol

Statin Drugs Ranges From 10-80 mg/day High Dosage are Given to Patients with High LDL Levels and Serum Cholesterol Levels Drug Dosage (mg/day) Percent Reduced Lipitor 10 to 20 30%-45% Mevacor 40 Pravachol Crestor 10 Zocor 20 to 40

How Statins Work How it Works: By inhibiting HMG-CoA in the Biosynthesis of Cholesterol Production Main Enzyme in Producing Cholesterol

Side Effects of Statins Common Side effects in most of the cholesterol lowering drugs Head pain Dizziness Rash Throwing up Some dissolve in fat and reach barriers of brain cell membrane Insomnia

Other Problems with Statins Interfere with the Biosynthesis of Coenzyme Q10 CoQ10 is Vitamin like, Fat-Soluble Antioxidant High Concentrations in Vital Organs like Heart Important in Energy Production Statins lower cholesterol in plasma and also CoAQ10 (Passi, 2003)

New Drug Squalestatin 1 Selective Inhibitor of Squalene Synthase Key Enzyme in Cholesterol Biosynthesis Because statins have high side effects, and lower efficacy when smaller doses are taken, a new solution that would lower the level but by using lower dosage and has lower side effects is very desirable.

New Drug SQ1 will be used to lower serum cholesterol level By Controlling the Flux of Cholesterol Biosynthesis Squalestatin 1 Farnesyl Pyrophosphatase

New Drug SQ1 Inhibits Squalene Synthase Lowers the Production of Cholesterol

How Dosage is Determined Inhibition dependant of daily production of squalene synthase (13.4 nmoles per day) squalestatin binds to squalene synthase to form complex Number Molecules Squalestatin = Number Molecules of Squalene Synthase Feze

Dosage 10 to 20 mg daily doses for 40 to 60% inhibition Feze

Dosage and Percent Reduction Drug Dosage(mg/day) % Reduced SQ1 20 60% Statins 40

Squalestatin Pathway Hepatic and Renal Elimination This is statins? Not squalestatins Feze

Renal Clearance Passive diffusion across glomerulus Fick’s law application P= permeability size dependant A= kidney filtration area (516 cm2) C= squalestatin concentration in the plasma and kidney Feze

Squalestatin 1 Clearance

Side Effects Side Effects: Have not been tested but it can be assumed lower side effects than other cholesterol lowering drugs because of its inhibition of a different enzyme.

Toxicology Evaluation Higher Clearance than Atorvastatins Lower doses 10 to 20 mg Vs 20 to 40mg Higher diffusions coefficient in the kidneys (shorter half life ) 8 hours Vs 14 hours

Toxicology Evaluation No Ubiquinone metabolism related side effects (43 % of severe side effect) Myalgia : muscle ache and weakness = 24% Arthralgia joint pain =19% Asthenia overall body weakness

Production Plan FDA process Manufacturing

FDA Approval Process Pre –FDA: Test tubes and mammalians testing Phase I: Safety test on small number of patients Phase II: Efficacy Test on Familial Hypercholesterolemia patients Phase III: Safety and efficacy tests on subjects with special conditions Hundreds to thousands of patients are volunteered for this phase. Last and most determining phase

Pre-FDA More tests = better rate of success through FDA Too many tests= waste of time and money

Pre-FDA Decision Tree 1 PhD, 8 labs Technicians PRE-FDA Option A, 28 Weeks $162,000 2 PhD, 12 labs Technicians 1 PhD, 12 labs Technicians Option B, 31 Weeks $179,000 Option A, 21 Weeks $162,000 Option B, 21 Weeks $162,000 Option A, 24 Weeks $185,000 Option B, 28 Weeks $215,000 Fix this one, try out everything in the middle

FDA Flow Chart (Phase I) 1year, 1.22millions, 70 subjects .1% 12.12% 87.78% No Antibody Stimulation Phase I Drug is Safe Adverse Side Effects Clinical Hold Up Back to R&D 30

Clinical Hold Up back to R&D Flow Chart (Phase II) 2 years, $5.2 millions ,200 subjects 1.1% 10.5% 10% 79.4% Phase II Severe Side Effects Minor Side Effects Drug is Ineffective Drug is Safe and effective Clinical Hold Up back to R&D

FDA Flow Chart (Phase III) 900 patients, $61.8 millions,6 years 0.50% 10% 89.5% Phase III Unexpected Side Effect Drug is Ineffective Booster to increase Titer Drug is safe and effective Biologics License Application Pre-approval Facility inspection Advisory Committee Clinical Hold Up Drug is not Safe Drug is Safe

FDA Summary Cost ($) In Millions Length Participants Phase I 1.22   Cost ($) In Millions Length Participants Phase I 1.22 1 year 70 Phase II 5.2 2 years 200 Phase III 61.8 6 years 900

FDA Summary The chances of passing are estimated at 69% The total cost is $69.3 millions for 10 years 31% Risk of failing FDA approval for a total loss of $138 millions

Manufacturing Process Calcium Salt Extraction

Squalestatin Extraction

Fermenter Scale Up H/D ratio Volumetric oxygen transfer coefficient (Kla) Power consumption volume Impeller tip velocity

Fermenter Scale-Up H/D ratio The most important factor in fermenter geometric design Advantages: bubble residence time increase top sparger air pressure is higher (higher O2 dissolution) Volume of air required decrease with constant SLV 38

Fermentor Scale-Up Air impelled fermenter Considerable mixing energy savings. Greater Kla value Greater sparger pressure

Column Chromatography scale-up Pressure drop across each column (Darcy equation) L= column height , ∆P= pressure drop across the column , µ = viscosity of the mobile phase, ν= superficial velocity K= constant, p= resin particle diameter, ε = void fraction, Column size V1,Q1 = pilot plant column volume and flow rate V2,Q2 = scaled up column volume and flow rate

Economics Price of the Drug Demand Model Production Manufacturing

Demand Model Assumptions Number of People Who Need a Type of Cholesterol Lowering Drug 8,000,000 800,000 Patients will Switch from the Highest Selling and the lowest Priced Statin Drug to this New Medication

Demand Model Assumptions Cond. Assume Percent Prescribed 10mg is prescribed 50% 20 mg is prescribed 25% 40 mg is prescribed 15% 80 mg is prescribed 10% Prescriptions are Renewed Every Year

Demand Model Equation  Awareness Function Function of time, advertising and professional education =0, No knowledge =1, Perfect Knowledge d1; Demand for the New Product ; Market Demand Assumed 0.78 Y; Consumer Budget P1 and P2 ; New and Old Product Price Alpha was varied when only the patients were taken into account as the customers and alpha was constant for only the doctors and health care professionals

Demand Model ; Consumer Satisfaction, Ratio of Old to New =H2/H1 H; Consumer Satisfaction Function  Wi yi y; Consumer Related Property Effectiveness; % serum cholesterol reduction Side effects; Related to Plasma Concentration of Drug W; Product Weight the consumer maximizes satisfaction by having constraints on the amount spent on the drug. P1d1+p2d2<=Y

Effectiveness Efficacy is the measure of the effectiveness of the drug in reducing the life threatening condition a patient is suffering from.

Effectiveness

Effectiveness Reduction in % Serum Cholesterol Level Lower=Higher Satisfaction By Measuring the Level in Blood

Side Effects Assumed that 50% of Prescriptions are for 10 mg/day No or Very Low Side Effects Less Important than the Efficacy of the Drug Based on a Doctor's Recommendation and Opinion

Beta Factor Beta Weight Y1 Y2 Efficacy 0.95 1.26 0.74 Side effects 0.05 0.5 H= (weight*y) 1.22 0.73  0.6

Alpha Factor Data taken from a previous study

Demand Model Different Scenarios, compared to other drugs,

Demand Model Price of the drug goes up, demand goes up

Price of the Drug Price of the New Drug $1.33 or $1.70 for 10 mg/day (one tablet) Demand model was solved for this range of values Demand below $1.33 did not make sense

Business Economics Equipment Prices

Business Economics Raw Material Suggested Factor Quantity Per year Cost Per Unit Value ($/year) Water (Process) $/Kg 70,339,560 0.53/1000 $37,280 Glycerol Kg 28,136 0.6 $16,881 Cotton Seed Flour 2,432 1 $2,432 Soybean Oil 84,407 2.5 $211,019 Actetonitrile 17,993,088 1.39 $25,010,392 Sulfuric acid 71,962 0.07 $5,037 Ammonium sulfate 450,944 0.08 $36,037 Resin liters 236 200 $47,124 Calcium acetate 172,486 179 $30,875,010 Phoma sp. 270,037 219 $59,138,059 Total $115,379,310 56

$271,041,780 Production Cost Suggested Factor Amount Per Year Price Per year Operating Labor 25.58$/h   $181,618 Operating Supervision 0.15 $27,243 Electricity 0.045$/kWh $18,696 Fuel 1.26$/GJ Steam 4.2$/1000kg 424 Kg $2 Maintenance 0.07 $4,691,327 Operating Supplies $703,699 Laboratory Charges $105,055 Utilities $5,594,778 Manufacturing Costs $121,107,449 Taxes 0.02 $1,340,379 Financing Insurance 0.01 $670,190 Rent Depreciation $3,015,853 Fixed Charges w/o Depreciation $5,026,422 Plant Overhead Costs 0.6 $144,907,909 Administrative Costs Included Above Distribution and Marketing Research and Development General Expenses Total Product Cost w/o depreciation $271,041,780

FCI/TCI FDA-Fixed One Time Cost $69,000,000 Fixed Capital Investment $76,018,956.73 Working Capital $1,239,458.63 Total Capital Investment $77,258,415.36 58

NPV NPV ($1.33) = $13,912,008,251 Based on NPV value and Demand Analysis, $1.33 and $1.70 are Best prices for SQ1 Projected is accepted because it is positive; 1 is the $1.33 and 2 is the $1.7

Risk Analysis The Risk of the Project Determined from NPV and Demand Graphs When the price is kept at 1.33 and the annual product cost changes over 20 years Assumed probabilities, based on the demand graph and the NPV versus time graph. Highest demand will in the years 4-14th of the project.

Conclusion High Cholesterol is a Growing Epidemic in the U.S. 20 mg/day of SQ1 will Reduce the Serum Cholesterol Level by 60% Higher Efficacy Lower Side Effects Demand for this Price of Drug is Close to 800,000 Patients Best Price to a Corresponding High Demand and NPV of the Project is $1.33 for 10 mg/day

References Baxter A., Fitzgerald B.J., Huston J.L., McCarthy A.D, Motteram J. M., Ross B. C., Sapra M., Snowden M.A., Watson N.S., Williams R.J., and Wright C., Squalestatin 1, a Potent Inhibitor of Squalene Sunthase, Which Lowers Serum Cholesterol in vivo, The journal of biological chemistry, Vol. 267, No. 17, Issue of June 15, pp. 11705-11708, 1992 Harl C. Erica, Martin Melissa, Financial and Technological Risk Analysis for the Development of New Drug, May 5, 2006 http://www1.istockphoto.com/file_thumbview_approve/2234414/2/istockphoto_2234414_sick.jpg http://apudgeisasandwich.files.wordpress.com/2007/08/money.jpg http://www.inkcinct.com.au/Web/CARTOONS/2006/2006-484-world-oil-demand.jpg http://www.naturalnews.com/cartoons/main_effects_600.jpg http://www.acc-tv.com/images/globalnews/hmed_pills_1106.jpg

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