Nanostructured Materials for Long Acting Biologic Delivery

Tejal A. Desai, PhDErnest L Prien Professor and ChairDirector, UCSF Engineering and Applied Sciences InitiativeDept. of Bioengineering and Therapeutic Sciences

7 February 2020

Conflicts

Zordera, Inc. VasaRx, Inc.

Therapeutic biomaterials for mitigating disease

• Drug Delivery and Targeting

• Cell Therapy and Immune Modulation

• Implants and Drug/Device combinations

We design and fabricate micro and nanomaterial solutions for:

The Desai Laboratory for Therapeutic Microtechnology and Nanotechnology

TopographyDevices

Thin FilmsNanoparticles

The Desai Laboratory for Therapeutic Microtechnology and Nanotechnology

Can we use material “structure” to achieve long

term biologic delivery?

Age-related Macular Degeneration

Major cause of vision impairment in elderly Significant visual impact to fine detail 2.07 million individuals affected in US

Normal Vision Degenerated Macula

National Eye Institute, Age-Related Macular Degeneration (AMD) Data and Statistics; www.nei.nih.gov

Wet-AMD Therapeutics – Disrupting VEGF

Lucentis (ranibizumab)~48kDa antibody fragment

0.5 mg injection monthly

First intraocular injection for wAMD

Binds and inhibits VEGF-A

Eylea (aflibercept)~115kDa fusion protein

After loading dose, 2 mg injection bimonthly

VEGF-R1 & VEGF-R2 extracellular domains + IgG1 Fc

Avastin (bevacizumab)~150kDa antibody

1.25 mg injection monthly

used off label

cost-effective alternative to Lucentis/Eylea

Clinical Need for Better Delivery Systems for wet-AMD

Current therapies all use intravitrealinjection- On average patients dosed 7.7 times per year- Peaks and troughs lead to poorer outcomes- Repeated injection results in risk of infection,

retinal detachment, and cataracts- Maximum dose is limited by inflammation

and elimination from eye

Requirements?- extended delivery duration (at least 4-6 mos)- A large drug depot- Syringe deployable- Device degrades at the end of use (no explant)

Concentration Dependent Delivery

“Single File”Constrained Delivery

Pore Size > Molecular Size Pore Size ~ Molecular Size

Achieving Constant Rate Delivery: Tuning the pore size

Modeling Device Pharmacokinetics

Continuous delivery can sustain therapeutic concentrations with equivalent payloads

Polycaprolactone Thin Films

Three core film types- Non-porous- Microporous (PEG porogen)- Nanoporous (ZnO Template)

27 January 2020

Device Fabrication

Can be flat, packet/sachet, cylindrical Load liquid, powder, compressed drug pellet

27 January 2020

Nanostructured films enable linear release kinetics for range of proteins

Bernards DA, 2012, Lance K et al, 2015

k0 ≈ 1.6 ug/day

Thin Film Devices for Eyelea (anti-VEGF)

Schlesinger et al., DDTR 2019

Ocular tolerability

• Post-implantation inflammation

- resolved 2-3 weeks

• No glaucoma or cataract formation seen

• Retina: no biomicroscopic or OCT abnormalities

Overall, intravitrealdevices well-tolerated over study period

McDonald-Shattuck score

IOP

mm

Hg

Flar

e ph

otom

etry

day

0da

y 28

day

56da

y 70

da

y 84

African green monkeys6 test eyes (drug device)8 placebo eyes (sucrose/PEG device)

Challenges in Glaucoma Drug Delivery

Studies show 50% of patients are not adherent over 75% of the time

Risk population (high age) have difficulty following dosing regimen and properly using eye droppers

1) Patient Compliance 2) Physiological Barriers

Robin et al (2011) & Zhang et al (2012)Layers of the cornea

(CRFA & Robertson et al, 2005)

Tight junctional complexes retard paracellular drug permeation

Kim K et al., Investigative ophthalmology & visual science 57 (10), 4341-4346; Kim J et al. Evaluation of polycaprolactone implant for long-term treatment of glaucoma: pilot study, JCR

In vitro

Glaucoma : intracameral device (DE-117)In vivo

Jean Kim 1

Cornea112244814262.283.96688.576.242.3Iris-ciliary body112244813634.915919228430086.4Vitreous humor11224481.554.260.691999999999999950.677000000000000051.421.39Retina-choroid11224487.81999999999999762.823.214.410.66.968.81

Time [weeks]

DE-117 [ng/g or ng/mL]

Intracameral Device for Glaucoma:Combination therapy: timolol and brimonodine

Timolol BrimonidineRelease rate (µg/day) 2.09 ± 0.04 0.60 ± 0.02Target rate (µg/day) 3.00 0.90

Scaling Down

What else can we do with thin film delivery?

Chiang N, Serhan CN. Mol. Asp. Med. 2015

Specialized Lipid Mediators (SPM) Drive Resolution

Vascular Effects of SPM

Reduced chemotaxis/migration of VSMC Reduce cytokine-stimulated leukocyte adhesion to EC, VSMC Reduce cytokine stimulated NF-kB activation and downstream

inflammatory gene expression Reduce cytokine stimulated ROS generation Modest anti-proliferative effects in VSMC

Ho KJ et al; Am J Pathology 2010 Akagi D et al. FASEB J 2015

Miyahara T et al ; FASEB J 2013 Wu B et al. J Vasc Surg 2016

Chatterjee A et al; PLoS One 2014 Lance K et al. J Biomed Mater Res A 2016

SPM-Eluting Biodegradable Film

Locally deliver SPM to sites of surgical/vascular injury to enhance repair

Developed a thin (e.g.

Sustained and directional elution of RvD1 from a biodegradable wrap

Directional ElutionSustained Elution

Wu B, et al J Vasc Surg 2016

Angio veh-gel

Perivascular delivery of SPM in an acute arterial injury model (rat)

Bypass Only Veh Gel RvD1 Gel Veh Wrap RvD1 Wrap

Wu B, et al. J Vasc Surg 2018; 68 (6S):188S-200S

Local RvD1 Treatment Reduces Vein Graft Hyperplasia -Rabbit Carotid Interposition Graft at 28 days

Can we design materials to modulate the local immune

environment long term?

Features vs. Challenges

Small signaling protein

Received interest in for

cancer & autoimmune

disease

Short half-life

Not bioavailable

Pleiotropic>30% require hospitalization

IFNγTNFαIL-2

Local Immune Activation via Cytokine Therapy

Endogenous cytokine complexes with

antibody

Localized, cell specific

activationCellular influx

Loose networkformation

Subcutaneous injection

T= 0 T= minutes

T= hours T= days

- Receptor A

- Receptor B

a b c d

Nanostructures as an injectable cytokine trap

Sprent et al., 2012

Can we use this strategy to activate T cells specifically and locally?

Conjugated Nanowire Fabrication

Zamecnik et al., ACS Nano 2017.

Scale Bar 20μm

45 kDa PCL is blended with short chain PCL containing a maleimide end groups

a

Lymph Node

Na

no

wi r

es

Ab

Na

no

wi r

es

Ab

on

l y

0

1 0

2 0

3 0

4 0

%K

i6

7+

N a t u r a l K i l l e r

* * * p < 0 . 0 0 0 1

Na

no

wi r

es

Ab

Na

no

wi r

es

Ab

on

l y

0

2 0

4 0

6 0

8 0

%C

D1

22

+

C D 8

* * p < 0 . 0 0 5 * p < 0 . 0 5

Na

no

wi r

es

Ab

Na

no

wi r

es

Ab

on

l y

4 0

5 0

6 0

7 0

8 0

%C

D2

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T r e g

Na

no

wi r

es

Ab

Na

no

wi r

es

Ab

on

l y

0

1 0

2 0

3 0

4 0

5 0

%K

i6

7+

N a t u r a l K i l l e r

Na

no

wi r

es

Ab

Na

no

wi r

es

Ab

on

l y

0

2

4

6

8

%C

D1

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+

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* p < 0 . 0 0 3

* p < 0 . 0 5

Na

no

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Ab

Na

no

wi r

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Ab

on

l y

4 0

4 5

5 0

5 5

6 0

6 5

%

CD

25

+

T r e g

SkinIL-2S4B6

Strong signal Weak signal

CD122 + CD25

CD8 Memory T cells, NK

Cells

Regulatory T Cells

CD122

S4B6 antibody-conjugated wires locally activate NK and CD8 Cells in vivo

Zamecnik et al., ACS Nano 2017.

T-reg specific nanowires for autoimmune applications

Zamecnik et al., Biomaterials 2019

Nanorods persist in vivo for >6 weeks

2 week 4 week 6 week

DAPI

F4/80

NWs

Scale bar -100μm

Decreased epithelial hyperplasia and myeloid infiltrate observed in vivo

Zamecnik et al., Biomaterials 2019.

Harnessing micro- and nanoscale cues for therapy

Acknowledgements

• NIH • NSF• JDRF• Thome Foundation• Al Mann Institute• CIRM• Eli Lily• SPARC

The Therapeutic Micro and Nanotechnology Laboratory

Nanostructured Materials for Long Acting Biologic DeliveryConflictsThe Desai Laboratory for Therapeutic Microtechnology and NanotechnologySlide Number 4Can we use material “structure” to achieve long term biologic delivery?Age-related Macular DegenerationWet-AMD Therapeutics – Disrupting VEGFClinical Need for Better Delivery Systems for wet-AMD Achieving Constant Rate Delivery: Tuning the pore sizeModeling Device Pharmacokinetics�Polycaprolactone Thin FilmsDevice FabricationNanostructured films enable linear release kinetics for range of proteinsThin Film Devices for Eyelea (anti-VEGF)�Ocular tolerabilityChallenges in Glaucoma Drug DeliveryGlaucoma : intracameral device (DE-117)Intracameral Device for Glaucoma:�Combination therapy: timolol and brimonodineScaling DownWhat else can we do with thin film delivery?Specialized Lipid Mediators (SPM) Drive ResolutionVascular Effects of SPMSPM-Eluting Biodegradable FilmSustained and directional elution of RvD1 from a biodegradable wrapPerivascular delivery of SPM in an acute arterial injury model (rat)Local RvD1 Treatment Reduces Vein Graft Hyperplasia -Rabbit Carotid Interposition Graft at 28 daysCan we design materials to modulate the local immune environment long term?Local Immune Activation via Cytokine TherapyNanostructures as an injectable cytokine trapCan we use this strategy to activate T cells specifically and locally?�Conjugated Nanowire Fabrication S4B6 antibody-conjugated wires locally activate NK and CD8 Cells in vivoT-reg specific nanowires for autoimmune applicationsNanorods persist in vivo for >6 weeks�Decreased epithelial hyperplasia and myeloid infiltrate observed in vivoHarnessing micro- and nanoscale cues for therapy�Acknowledgements