CLICK PEPTIDIC TO COMPOUND EDIT MASTER FOR IMPROVING TITLE STYLE EFFICIENCY OF TRANSFECTION 22. May 2017 Bayerische Patentallianz LMU Joseph Rosenecker Dr. von Haunersches Kinderspital Ludwig-Maximilians-Universtität München Forschungszentrum Kubus
Field of interest Gene Therapy Advantage: promising therapeutic strategy for many lung diseases intractable to traditional treatments, e.g. Cystic fibrosis (CF) Problem: delivery issuses caused by extra- and intracellular barriers Poloxamine 704 (T704) T704 based nonviral carriers Advantages: More efficient than cationic lipids or polymer in vivo Low inflammatory-causing properties Problems: Invalid for in vitro transfection in vivo transfection efficiency still low compared with viral vectors GL6 7A x=14, y=13 T70 4 Richard-Fiardo et al., 2015, Biomaterials (45), 10-17 22. May 2017 Bayerische Patentallianz LMU 2
Peptidic compound Anchor moiety Cationic moiety Targeting moiety Candidate module -Lipoic acid - Hydrophobic amino acids -Fatty acid CLICK TO EDIT MASTER -LHHHKKLLHHKKL TITLE STYLE -Cholesteryl Candidate module -KKRRRRKK -KKKKRRRRRKKKK -KKRRRRRRRRRKK -KKRRRRRRRRKK -KRRRRK Candidate module -PKKKRKV -GACSERSMNFCG -GACYGLPHKFCG - Interact with T704 - Protect nucleic acids - Endosome escape - Increase specific cellular uptake - Organells targeting (e.g. nucleus) Other candidate modules can be tailor-designed according to target tissue or specific requirement 22. May 2017 Bayerische Patentallianz LMU 3
Peptidic compound helps to overcome intracellular barriers pdna Binary complex: + pdna pdna Ternary complex: + + pdna mrna Ternary complex: Self-assemble + + IVT-mRNA Self-assemble 22. May 2017 Bayerische Patentallianz LMU 4
Peptidic compound enables efficient in vitro transfections of T704 1) pdna (luciferase) transfection: 2) Peptidic-T704 nanocomplex also mediate efficient IVT-mRNA (EGFP) transfection: pdna Binary complex IVT-mRNA pdna Ternary complex: 22. May 2017 Bayerische Patentallianz LMU 5
Peptidic compound enables more potent pulmonary transfection of T704 NF-water DOTAP lipoplex Binary complex Ternary complex 22. May 2017 Bayerische Patentallianz LMU 6
Peptidic compound also works fine with other poloxamines and poloxamers Poloxamine 304 Poloxamine 904 Poloxamine 90R4 Poloxamer 184 Without peptidc compound, all these amiphiphilic block copolymers just mediate backgroud levels of in vitro transgene expression 22. May 2017 Bayerische Patentallianz LMU 7
Future Development For T704-Peptidic ternary complex Further optimization of peptidic compound structure with other functional candidate modules Test T704-Peptide ternary comlex in transfecting other tissues (e.g. muscle and myocardial) Test T704-Peptide ternary comlex in large animal models (e.g. Porcine and primate) Investigate T704-Peptide ternary comlex in mediating genome editing for long-term expression For other Poloxamines and Poloxamers Test and optimization of peptidic compound with other Poloxamines and Poloxamer Evaluate the in vivo transfection efficiency of promising candidates Test with other potential amphiphilic block copolymers For Vaccine applications 22. May 2017 Bayerische Patentallianz LMU 8
Innovation Huge unexplored potentials Unlike the cationic lipids or polymers which have been invstigated for decades, Poloxamines and Poloxmers are emerging materials for gene transfer applications Easy to prepare and low-cost All necessary materials in current patent have been produced in GMP grade with low price Versatile The structure of peptidic compound can be tailor-designed for different purposes (e.g. targeting to different tissues or organells) Extensive applications peptidic compound strategy may be generalizable to all Poloxamines and Poloxmers which has been evaluated in many in vivo studies but were invalid for in vitro application 22. May 2017 Bayerische Patentallianz LMU 9
IP status & commercial potential IP status PCT application 03/2017 Commercial potential In-vitro transfection reagent Clinical application Gene Therapy Vaccination 22. May 2017 Bayerische Patentallianz LMU 10
Thank you for your attention Meet us at Poster B1 Scientific expertise Commercialization Prof. Dr. Joseph Rosenecker Dr. Jens Nagel Shan Guan +49 89 5480177-16 jnagel@baypat.de 22. May 2017 Bayerische Patentallianz LMU 11