Index Introduction Experiment Results Conclusion

Introduction 2004 Summer San Francisco State University Dr. Clifford Berkman’s lab Research Purpose: To make the medicinal drug for prostate cancer.

Introduction Prostate cancer is one of the most frequently diagnosed cancers among men and the second most common cause of cancer-related deaths in American (second to lung cancer). About 200,000 men are diagnosed each year. Reference: Prostate.com

Introduction: Prostate Cancer Benign or Malignant Malignant cancer Cells Spread from the prostate to nearby lymph nodes, bones or other organs.

Introduction: PSMA When A man has prostate cancer, an enzyme called PSMA is located on the surface of the cancer cell PSMA: Prostate Specific Membrane Antigen Enzyme is a Protein used to catalyze biological reactions The Enzyme PSMA

What ’ s going on at the Surface Domain E is responsible for the catalytic activity of the enzyme. The other Domains has no enzyme activity E: Catalytic activity A model for the organization of the PSMA. Prostate Cancer Cell

Introduction: Inhibitors The Medicinal Drugs we were trying to make this summer are called inhibitors Inhibitors are compounds that stop the growth of cancer cells Preliminary Studies done by Dr. Berkman suggest that compounds that look like the one in figure 1, can decrease cancer growth Figure 1

Introduction X = 4-Cl, 4-OMe, 4-Me, 3,4-Cl2, 4-OPh 3-Cl, 4-tBu, 4-CF3, 4-Br, 3-CF3-4-Cl, 3-Me, 4-N(Me)2, 3-N(Me)2, 3-CF3, 2,4-Cl2, 4-NO2, 3-CF3-4-NO2, 4-NH2, 4-OH, 3-Ome, 4-OiPr, 2-Cl, 2-Me, 2-OMe, 4-F, 3,5-Cl2, 3-NO2, 4-OCH2Ph, Figure 1 X could be those following molecules. The following molecule looks like the drug we are trying to design for prostate cancer.

Introduction In order to make the compound in Figure 1, the first step is to make the compound in Figure 2. Figure 2 Figure 1

Wanted and Unwanted Compound Wanted Compound Unwanted Compound During the course of the summer, my responsibility was to make the desired molecule above. But we had a lot of problems with the chemistry. Our Desired compound kept isomerizing into the unwanted compound above.

The Experimental The next couple of slides details how the Wanted and Unwanted compounds were made

Experimental: Experiment 1 Reaction Scheme

Experiment Step 1 Add Dibenzylphosphite to a round bottom flask with a magnetic stir bar. Set this flask on an Ice Bath Add Potassium hexamethyl disilazane ( KHMDS) drop wise and stir for 1hour Step 2 Remove the Ice Bath Add allyl bromide drop wise and stir overnight At this point the heat conditions will be employed Experiment 1

Experiment Experiment 2 …Resaction Scheme Dibenzylphosphite Side Product Allylbromide Phosphonate Salt Potassium tert-Butoxide

Experiment Step 1 Add Dibenzylphosphite to a round bottom flask with a magnetic stir bar. Set this flask at room temperature Add Potassium tert-Butoxide (powder) and stir for 1 hour Step 2 Add Allyl Bromide drop wise and stir overnight At this point the heat conditions will be employed Experiment 2

Experiment Using a Separatory Funnel, the reaction mixture inside Add Ethyl Acetate (EtOAc) Add water There were two layers present. organic was at the top aqueous was at the bottom Shake the separotory funnel and release aqueous layer Wash with 10% HCl and shake and release aqueous layer Wash with NaHCO 3 and shake and release aqueous layer Wash with Sat. NaCl and shake and release aqueous layer Dry the reaction with Na 2 SO 4 Filter Na 2 SO 4 and evaporate the solvent Work up Procedure (For Both Experiments)

Experiment Once the solvent is evaporated, Run a TLC using 10% Isopropanol in Hexane 3% Methanol in Dicholoromethane Run a Crude NMR Purification with 3% Methanol in Dichloromethane Run a Pure NMR Get the graphs of NMR tests. Test Procedure

Results Experiment 1 gave both wanted and unwanted product. Experiment 2 gave only the desired product. Comparison The Wanted Compound Allylphosphonate The Unwanted Vinylphosphonate

Results Experiment 1% Yields Temperature o C Major CompoundStarting Materials Wanted Compound Unwanted Compound 0Unwanted Product0%8%92% 25Unwanted Product0%14%86% 40Unwanted Product0%10%90% The results of the experiment 1 gave the unwanted compound and a small amount of the wanted compound. Experiment 1

Results Experiment 2 The results of the experiment 2 gave the majority of the wanted compound and a small amount of the starting material. No unwanted product was present. Experiment 2% Yields TemperatureResulting Compound Starting Materials Wanted Compound Unwanted Compound 25Wanted Product 25%75%0%

Results The reason why I tried experiment 1 under three different conditions is at each temperature condition the major product was the unwanted compound. By changing the condition we thought that we could produce the wanted compound. We tried experiment 2 only one time and received the wanted compound with the first attempt.

NMR Results The next couple of slides Characterize the Compound found in Experiment 2

Ethyl Acetate HbHb Ha This a crude NMR The Crude NMR is shown to show that the reaction did not produce any of the unwanted compound There is some allylbromide present. And some EtOAc present but we know that the wanted compound is present because of the Ha and Hb Peaks. Benzene Ring Region Proton NMR test result of an experiment after experiment 2

Results Phosphorus NMR test result of an experiment after experiment 2 This test looks like we get the wanted compound ppm indicates our wanted compound.

Conclusion The wanted compound may not be stable when using the conditions from Experiment 1, because we saw it isomerizing to the unwanted compound. The wanted compound was formed when we tried Experiment 2. This experiment will be done to conform the results. Other chemical methods will be used for making the allylphosphonate.

Special Thanks Dr. Berkman Dr. Berkman Thanks for helping and teaching Mentor: Nichole Coleman Mentor: Nichole Coleman Thanks for helping and teaching me patiently. Ms. Elaine Yamaguchi & Mr. Glann Fuller & Wally Yokoyama Ms. Elaine Yamaguchi & Mr. Glann Fuller & Wally Yokoyama American Chemical Society American Chemical Society Mr. Matsumoto Mr. Matsumoto Project Seed Mentors Project Seed Mentors Thanks for helping me. Project Seed Students Project Seed Students Thanks for having lunch with me.^-^

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