Small Molecule-DNA Hybrids (SMDHs) for Cancer Therapy Andrew Kim Nguyen Group PI: Prof. SonBinh T. Nguyen Mentor: Dr. Bill Hong Northwestern University, Posner Fellowship

Outline Cancer and Current Treatments Ideal Cancer Therapy Agent Small-Molecule DNA Hybrids (SMDHs) Synthesis of SMDHs Future Work

Cancer and Current Treatments 1.5 million new cases  500 thousand people Major types of treatment include: Surgery Chemotherapy I think you are going to emphasize ‘targeted therapy” in this slide, but you should think of how you well connect this with the next slide “ideal cancer therapy agent”? https://www.cancer.gov/about-cancer/understanding/statistics

Ideal Cancer Therapy Agent What characteristics does the ideal cancer therapy agent have? Targeting Agent – Guides drug to specific cancer cells Multi Drug Molecules – Give synergy effect for cancer treatment I think the left and the right should be switched as considering the small title. You should ask me how to change these things tomorrow. Tracking Agent  Targeting Agent (I think targeting agent is correct, but you should double check) Specific cells  specific cancer cells Drug molecules  Multi drug molecules; Anticancer drug payload  kill cancer cells Visual response molecule  monitor drug response Imaging Agent – Monitors drug response Multifunctionality

One of the Best Cancer Therapy Agents so Far Antibody-Drug Conjugate (ADC) Biodegradable Linking Molecule Targeting Agent (Antibody) Drug Molecule Change the title: this is not ours. e.g.) Emerging targeted therapeutics Tracking agent Targeting agent (https://adcreview.com/adc-university/adcs-101/how-do-antibody-drug-conjugates-work/)

ADC vs. Ideal Cancer Therapy Agent Targeting Agent Targeting Agent Drug Molecule #1 Drug Molecule #1 Drug Molecule #2 It is better to place “SMDH” and “ADC” on each box, respectively. And remove the text below the right box unless it is necessary. Tracking agent Targeting agent Imaging Agent

Potential Solution for Drug Delivery Small Molecule-DNA Hybrids (SMDHs) – DNA is conjugated onto a small molecule The title does not explain the content in this slide. Think of what you are going to tell in this slide and what title should be? Before adding other entities conjugated complementary DNA strands, SMDH has a single strand DNA (ssDNA), not double strand DNA (dsDNA). Find a figure of ssDNA and replace it with dsDNA.

Potential Solution for Drug Delivery Multifunctionality: Targeting agent, Imaging agent, other drugs Drug 2 Drug 1 Same as the previous slide. Change the title. In this slide, dsDNA is correct since complementary DNAs conjugated with other entities hybridize with the ssDNA of the SMDH. But, in your animation, you should show ssDNA first, then dsDNA with other entities. Drug  Drug 1; Drug  Drug 2 Tracking agent Targeting agent Targeting Agent Imaging Agent

Potential Solution for Drug Delivery Multifunctionality: Targeting agent, Imaging agent, other drugs Drug 2 Drug 1 Same as the previous slide. Change the title. In this slide, dsDNA is correct since complementary DNAs conjugated with other entities hybridize with the ssDNA of the SMDH. But, in your animation, you should show ssDNA first, then dsDNA with other entities. Drug  Drug 1; Drug  Drug 2 Tracking agent Targeting agent Targeting Agent Imaging Agent

Research Goal Goal: Modulate the DNA arms on SMDH. SMDH3 SMDH2 SMDH1 I don’t understand this figure. You should explain me tomorrow. SMDH3 SMDH2 SMDH1

Research Goal Goal: Modulate the DNA arms on SMDH. Problem with a previous synthesis method The right illustration and the left HPLC profile should be switched. SMDH4 SMDH2 SMDH1 SMDH3 Limiting factor: no way to control the DNA arms Chem. Sci., 2014, 5, 1091

By adding protecting groups, the DNA arms on SMDHs can be controlled Solution = Protecting Group = DNA strand By adding protecting groups, the DNA arms on SMDHs can be controlled

Outline Cancer and Current Treatments What are Antibody-Drug Conjugates (ADCs)? Why use small-molecule DNA Hybrids (SMDHs)? Synthesis and Protection of SMDHs Future Work

Step 1 – Synthesis of Tetrakis(4-cyanophenyl)methane CuCN DMF, 160⁰C, 4h Tetrakis(4-bromophenyl)methane Tetrakis(4-cyanophenyl)methane Hoskins, B. F, et al. J. Am. Chem. SOC. 1990, 112, 1546-1554

Step 1 – Synthesis of Tetrakis(4-cyanophenyl)methane 2 1 2 Tetrakis(4-cyanophenyl)methane

Step 2 – Synthesis of Tetrakis((4-aminomethyl)phenyl)methane Tetrakis(4-cyanophenyl)methane Tetrakis((4-aminomethyl)phenyl)methane Gandhamsetty, N., et al. J. Org. Chem. 2015, 80, 7281−7287

Protection of Core Fluorenylmethyloxycarbonylchloride (Fmoc) - commonly used to protect amine group + Protected Amine Group

Solid Phase DNA Synthesis DNA synthesizer https://en.wikipedia.org/wiki/Oligonucleotide_synthesis#/media/File:H-Phosphonate_Cycle.tif

Solid Phase SMDH synthesis DNA conjugated onto open branches + Open branches

Future Work Remove protecting group and conjugate different DNA strands onto SMDH Small-Molecule DNA Hybrids have the potential to be developed further towards multifunctional cancer therapeutic agents.

Acknowledgements Mentors: Prof. SonBinh Nguyen Bill Hong, Ph.D. Nguyen Group Northwestern University Posner Fellowship

Potential Solution for Drug Delivery Options to vary functional molecules Drug Drug Same as the previous slide. Change the title. If you want to emphasize in this slide that more drugs can be delivered by a single SMDH, change the small title. Do not just copy and paste the previous one. Targeting Agent Drug