ADC- Creation of Cytotoxic Payload Poster

Objective Fig 1: Standard Antibody Drug Conjugate Fig 7: LC/MS of conjugate, product impure with traces of doxorubicin and linker. M/z ratio of product 1142.6, linker 736.5 and doxorubicin 544.3 Fig 6: Thin Layer Chromatography (TLC) Plate Doxorubic in Fig 2: ADC Mechanism of Action Fig 3: Doxorubicin Topoisomerase II Inhibition Fig 4: Silica Column Linker Attachment Group Val Cit Space r Fig 5: Schematic of drug and linker conjugated product Fig 8: Attachment group conjugated to Trastuzumab Introduction Materials and Methods Results Conclusion Acknowledgement s References Future Direction Rf Values Dox: 0 (Dox) Dox+Rxn Mixture: 0.5 (Conjugate) Rxn Mixture: 0.5 (Linker) Background Linker Dru g Synthesis of a Novel Linker, MC-val-cit-PAB-PNP, for Payload of ADC Zoe Vaughn, Alexandra Van Hall, Dr. Dhavalkumar Shah Department of Pharmaceutical Sciences, University at Buffalo, Buffalo, NY • Measure the effectiveness of the linker, doxorubicin conjugate via conjugation to antibody, Trastuzumab • Create an ADC with linker, doxorubicin and Trastuzumab conjugate • Test effectiveness of ADC via cultured cancer cells • TLC plate indicates conjugate in fraction 6, with Rf value of 0.5 (Fig 6) • LC/MS indicates impure conjugate with m/z value of 1146.2 consistent with [4] (Fig 7) 1. Ducry, Laurent, and Bernhard Stump. "Antibody-Drug Conjugates: Linking Cytotoxic Payloads to Monoclonal Antibodies." Bioconjugate chemistry 21.1 (2010): 5-13. Print. 2. Jain, Nareshkumar, et al. "Current Adc Linker Chemistry." Pharmaceutical research 32.11 (2015): 3526-40. Print 3. Francisco, Joseph A., et al. "Cac10-Vcmmae, an Anti-Cd30– Monomethyl Auristatin E Conjugate with Potent and Selective Antitumor Activity." Blood 102.4 (2003): 1458-65. Print. 4. Dubowchik, Gene M., et al. "Cathepsin B-Labile Dipeptide Linkers for Lysosomal Release of Doxorubicin from Internalizing Immunoconjugates: Model Studies of Enzymatic Drug Release and Antigen-Specific in Vitro Anticancer Activity." Bioconjugate Chemistry 13.4 (2002): 855-69. Print. Antibody drug conjugates (ADC’s) are gaining popularity due to their ability for targeted cancer cell therapy which lessens adverse side effects to normal tissue. A suitable payload and linker antibody combination is very important for successful targeted therapy. [1] • Shah Lab • National Science Foundation LC/MS used to identify the continence of the purified product (Fig 7) TLC plates used to identif y purifie d product . Conjuga te present in fractio n 6 (Fig 6) Silica column created (Fig 4). Diluted product purified through column 7 fractions collected Completed reaction product frozen and centrifuged crashing out solid product Stir reactants in dark for approximatel y 2 days Reactants : MC-Val- Cit-PAB- PNP linker, Doxorubic in, DIEA base, HOBt catalyst To synthesize a novel linker, MC-val-cit-PAB-PNP, for payload of ADC.

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Transcript of ADC- Creation of Cytotoxic Payload Poster

Page 1: ADC- Creation of Cytotoxic Payload Poster

Objective

Fig 1: Standard Antibody Drug Conjugate

Fig 7: LC/MS of conjugate, product impure with traces of doxorubicin and linker. M/z ratio of product 1142.6, linker 736.5 and doxorubicin 544.3 Fig 6: Thin Layer Chromatography (TLC) Plate

Doxorubicin

Fig 2: ADC Mechanism of Action

Fig 3: Doxorubicin Topoisomerase II Inhibition

Fig 4: Silica Column

Linker

Attachment Group Val CitSpacer

Fig 5: Schematic of drug and linker conjugated product

Fig 8: Attachment group conjugated to Trastuzumab

Introduction Materials and Methods

Results

Conclusion

Acknowledgements

References

Future Direction

Rf ValuesDox: 0 (Dox)Dox+Rxn Mixture: 0.5 (Conjugate)Rxn Mixture: 0.5 (Linker)

Background

Linker Drug

Synthesis of a Novel Linker, MC-val-cit-PAB-PNP, for Payload of ADC

Zoe Vaughn, Alexandra Van Hall, Dr. Dhavalkumar ShahDepartment of Pharmaceutical Sciences, University at Buffalo, Buffalo, NY

• Measure the effectiveness of the linker, doxorubicin conjugate via conjugation to antibody, Trastuzumab

• Create an ADC with linker, doxorubicin and Trastuzumab conjugate

• Test effectiveness of ADC via cultured cancer cells

• TLC plate indicates conjugate in fraction 6, with Rf value of 0.5 (Fig 6)

• LC/MS indicates impure conjugate with m/z value of 1146.2 consistent with [4] (Fig 7)

1. Ducry, Laurent, and Bernhard Stump. "Antibody-Drug Conjugates: Linking Cytotoxic Payloads to Monoclonal Antibodies." Bioconjugate chemistry 21.1 (2010): 5-13. Print.

2. Jain, Nareshkumar, et al. "Current Adc Linker Chemistry." Pharmaceutical research 32.11 (2015): 3526-40. Print

3. Francisco, Joseph A., et al. "Cac10-Vcmmae, an Anti-Cd30–Monomethyl Auristatin E Conjugate with Potent and Selective Antitumor Activity." Blood 102.4 (2003): 1458-65. Print.

4. Dubowchik, Gene M., et al. "Cathepsin B-Labile Dipeptide Linkers for Lysosomal Release of Doxorubicin from Internalizing Immunoconjugates: Model Studies of Enzymatic Drug Release and Antigen-Specific in Vitro Anticancer Activity." Bioconjugate Chemistry 13.4 (2002): 855-69. Print.

Antibody drug conjugates (ADC’s) are gaining popularity due to their ability for targeted cancer cell therapy which lessens adverse side effects to normal tissue. A suitable payload and linker antibody combination is very important for successful targeted therapy. [1]

• Shah Lab • National Science Foundation

LC/MS used to identify the continence of the purified product (Fig 7)

TLC plates used to identify purified product. Conjugate presentin fraction 6 (Fig 6)

Silica column created (Fig 4). Diluted product purified through column 7 fractions collected

Completed reaction product frozen and centrifugedcrashing out solid product

Stir reactants in dark for approximately 2 days

Reactants: MC-Val-Cit-PAB-PNP linker, Doxorubicin, DIEA base, HOBt catalyst

To synthesize a novel linker, MC-val-cit-PAB-PNP, for payload of ADC.