Nanocarriers from GRAS Zein Proteins to Encapsulate Hydrophobic

Oct 16, 2016 - Badal Kumar BiswalMahmoud El SadanyDivya KumariPoonam SagarNitin Kumar SinghalSandeep SharmaTsering StobdanVijayakumar ...
0 downloads 0 Views 2MB Size
Subscriber access provided by UNIVERSITY OF LEEDS

Article

Nanocarriers from GRAS Zein Proteins to Encapsulate Hydrophobic Actives Nikolas T. Weissmueller, Hoang Dung Lu, Amanda Hurley, and Robert K Prud'homme Biomacromolecules, Just Accepted Manuscript • DOI: 10.1021/acs.biomac.6b01440 • Publication Date (Web): 16 Oct 2016 Downloaded from http://pubs.acs.org on October 18, 2016

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Biomacromolecules is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Biomacromolecules

1

Nanocarriers from GRAS Zein Proteins to Encapsulate

2

Hydrophobic Actives

3 4

Nikolas T. Weissmueller†1, Hoang D. Lu†1, Amanda Hurley2, Robert K. Prud’homme*1

5

1. Department of Chemical and Biological Engineering, Princeton University, Princeton, New

6

Jersey 08544, United States.

7

2. Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, United

8

States.

9 10

Keywords:

11

Nanoparticle, zein, casein, autoinducer, drug-delivery, GRAS, cholera

12 13

Abstract

14

One factor limiting the expansion of nanomedicines has been the high cost of the materials and

15

processes required for their production. We present a continuous, scalable, low cost nano-

16

encapsulation process, Flash Nanoprecipitation (FNP) that enables the production of nanocarriers

17

(NCs) with a narrow size distribution using zein corn proteins. Zein is a low cost, GRAS protein

18

(having the FDA status of “Generally Regarded as Safe”) currently used in food applications,

19

which acts as an effective encapsulant for hydrophobic compounds using FNP. The four-stream

20

FNP configuration allows the encapsulation of very hydrophobic compounds in a way that is not

21

possible with previous precipitation processes. We present the encapsulation of several model

22

active compounds with as high as 45 wt% drug loading with respect to zein concentration into

23

~100 nm nanocarriers. Three examples are presented: (1) the pro-drug antioxidant, vitamin E-

ACS Paragon Plus Environment

1

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 2 of 29

24

acetate, (2) an anti-cholera quorum-sensing modulator CAI-1 ((S)-3-hydroxytridecan-4-one).

25

CAI-1 that reduces Vibrio cholerae virulence by modulating cellular communication, and (3)

26

hydrophobic fluorescent dyes with a range of hydrophobicities. The specific interaction between

27

zein and the milk protein, sodium caseinate, provides stabilization of the NCs in PBS, LB

28

medium, and in pH 2 solutions. The stability and size changes in the three media provide

29

information on the mechanism of assembly of the zein:active:casein NC.

30

Background

31

Nanomedicines: Nanotechnology has been an area of intense research commitment over

32

that last two decades, and nanocarriers (NCs) for the delivery of therapeutics has been one of the

33

successful areas in biomedical nanotechnology. Most successes in NC delivery have been in

34

oncology, where the high value of treatment has allowed the application of relatively expensive

35

formulations. Formulations most often employ block copolymers with polyethylene glycol

36

(PEG) blocks to provide biocompatible surface properties.1-2 NCs for oral administration, most

37

often, have as their aim increased bioavailability of very hydrophobic drug compounds.

38

formulations must balance the advantage of increased bioavailabilty against the increased costs

39

of NC processing and of the excipients required for NC formation. Using materials that are

40

accepted by regulatory agencies as GRAS (Generally Regarded as Safe) are preferred in oral

41

delivery applications, as they reduce the complexity of regulatory approval.

3

Oral

42

Our interest is in the encapsulation of hydrophobic compounds for oral delivery using

43

low-cost, GRAS excipients. Biodegradable protein polymers, such as albumin, casein, gelatin,

44

and chitosan, have been investigated as all-natural low-cost encapsulants.

45

soluble proteins generally provide poor coupling with hydrophobic compounds and, therefore,

46

result in low loading and low encapsulation efficiency. Excellent work by the Johnston and

ACS Paragon Plus Environment

4

However, these

2

Page 3 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Biomacromolecules

47

Elder’s group on direct precipitation of hydrophobic drugs with hydroxypropyl methylcellulose

48

(HPMC) has demonstrated significant increases in drug supersaturation upon dissolution of the

49

resulting powders.5-8 Direct precipitations rely upon highly hydrophobic compounds to achieve

50

high nucleation rates, and encapsulation by HPMC relies solely on hydrophobic interactions

51

between the polymer and compounds. Both hydrophobic and electrostatic interactions between

52

zein and the compounds being encapsulated provides increased flexibility in compounds that can

53

be processed into NC form.

54

In this paper we present the new process for the encapsulation of hydrophobic actives

55

with zein proteins using the kinetically controlled, rapid precipitation process Flash

56

NanoPrecipitation (FNP) using a Multi-inlet Vortex Mixer (MIVM). The MIVM, with multiple

57

inlet streams enables the encapsulation of actives at higher loading, control of size, and narrow

58

size distributions than have been reported for alternate NC formation processes. The specific

59

interaction between zein and the milk protein casein provides effective stabilization of the NCs.

60

We provide three examples to show the generality of the process: (1) the pro-drug antioxidant,

61

vitamin E-acetate, (2) an anti-cholera quorum-sensing modulator CAI-1 ((S)-3-hydroxytridecan-

62

4-one) and (3) hydrophobic fluorescent dyes with a range of hydrophobicities.

63 64

Zein prolamin proteins: In this study we present NC formation based on rapid

65

precipitation with zein as the encapsulating agent. Zein is a low cost, GRAS prolamin protein

66

found in the endoplasmic reticulum-derived protein vesicles in maize seeds.9 It finds application

67

as a film coating excipient of pharmaceuticals. Zein is water-insoluble owing to its high content

68

(>50%) of non-polar amino acids such as leucine, proline, alanine, and phenylalanine.10 It is also

69

insoluble in pure alcohol or most organics. It is soluble in water:alcohol mixed solvents, which

ACS Paragon Plus Environment

3

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 4 of 29

70

provides a route for processing zein into water insoluble particles by mixing with excess water.

71

However, this presents a barrier for incorporating highly hydrophobic actives and zein in simple

72

mixing/phase separation processes because the highly hydrophobic active will not be soluble in

73

high enough concentrations in an alcohol:water mixture to ensure adequate active loading.

74

Globular zein consists of four fractions that vary in molecular weight, composition,

75

structure and solubility.9, 11 These include α-zein (MW, 19–24 kDa; 75–80% of total protein) 12,

76

β-zein (17–18 kDa, 10– 15%), γ-zein (27 kDa, 5–10%), and δ-zein (10kDa)

77

subcomponents are arranged into a tertiary structure that comprises nine homologous repeating

78

units oriented in an anti-parallel sense, and stabilized by hydrogen bonds.11 The majority of the

79

molecular surface area comprises the hydrophobic α-helixes in anti-parallel orientation 16, while

80

the glutamine rich turns create a hydrophilic surface at their top and bottom.

81

assembly bestows zein with amphiphilic characteristics. These properties are reported to drive

82

self-assembly into a variety of mesostructures, 17 including ribbons, sheets, tori, pores, and micro

83

and nanospheres15. The amphilicity of zein allows it to encapsulate a variety of biological

84

compounds: heparin

85

D3

86

thymol

87

unique, and points to the encapsulation being driven both by hydrophobic and electrostatic

88

interactions. A limitation has been that reported loading capacities are generally 98% for all stable formulations as determined by absorbance measurement of the flow through

394

after centrifugal filtration (SI Figure 1). With increasing Nile Red concentration in the core, the

395

mean NC diameter increases (Fig. 7b). The florescence intensity increased with higher dye

396

loading per particle. This can be seen in Fig. 7c and 7d for two different wavelengths (500nm

397

excitation, 690nm emission) and core loadings of 0.2-1% wt. However, at dye content higher

398

than 1%wt. the fluorescence intensity decreases, as intermolecular interactions lead to

399

quenching.61

ACS Paragon Plus Environment

20

Page 21 of 29

A

B

Nile Red Pyrene Methyl Red

16 14

NP Mean Diameter

106

Size (d.nm)

Intensity (%)

108 107

12 10 8 6 4

105 104 103 102 101

2 100 0 100

1000

10000

0

1

2

Size (d.nm)

400

C

3

4

5

Nile Red % wt.

D 1400

3500

Excitation Maximum

Emission Maximum

3000 1200

Fluorescence

2500

Fluorescence

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Biomacromolecules

2000 1500 1000 500

1000 800 600 400 200

0

0 0

1

2

3

4

5

6

0

1

Nile Red % wt.

401

2

3

4

5

Nile Red % wt.

402

Figure 7: Characterization of NC loading. (A) Sizes of dye-containing Zein:CAS:VitE-Ac NCs by dynamic light

403

scattering. Formulations contained 2 mg/mL zein in 60% EtOH at 12mL/min, flashed with 1mg/mL casein in

404

sodium citrate buffer (36mL/min), flashed with 0.33 mg/mL VitE-Ac and 0.01wt % of dye in 100% EtOH at

405

12mL/min. (B) Mean diameter size of NC formulations with Nile Red loading from 0.25-5% wt. Fluorescence

406

maxima for either (C) 500nm excitation, or (D) 690nm emission, at 0.25-5% wt. Nile Red loading for

407

Zein:CAS:VitE-Ac NCs.

408 409 410

Conclusion

ACS Paragon Plus Environment

21

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 22 of 29

411

Zein protein-based NC formulations were prepared using sodium caseinate (CAS) as a

412

stabilizer. Particles at sub-100 nm sizes, with high loading of highly hydrophobic components

413

such as VitE-Ac, CAI-1, and hydrophobic dyes is enabled by the ability to form NCs with a

414

multi-inlet vortex mixing geometry (MIVM) that enables independent control over multiple inlet

415

streams. The amphiphilic molecular structure of zein enables both assembly of the NCs, and also

416

synergistic interactions with casein proteins to produce stable NCs. The components in the

417

formulations are all GRAS, which means that translation to food and oral drug delivery

418

therapeutics is facilitated. The dramatically lower cost of zein and casein, relative to the

419

amphiphilic block copolymers previously used in block copolymer stabilized NCs, expands the

420

range of applications that will be of interest. The precise NC size control, and the scalability of

421

the FNP process suggests applications in food (e.g. nutraceutical), pharmaceutical and

422

agricultural formulations.

423

Encapsulation of the quorum sensing therapeutic, CAI-1 demonstrated that the NCs

424

remained colloidally stable in ionic buffers and simulated intestinal fluid for 24 hrs. Efficacy of

425

CAI-1 in NCs was similar to CAI-1 in DMSO, while enabling CAI-1 delivery as an aqueous

426

dispersion. Within the context of cholera, the low-cost GRAS components and the scalability of

427

the FNP process may enable an auxiliary prophylactic treatment that could limit the need for

428

antibiotics.

429

The encapsulation of both purely hydrophobic compounds (VitE-Ac, CAI-1) has been

430

demonstrated by FNP previously. The encapsulation of the ionizable methyl red and Nile Red

431

point out the interesting ability of zein to interact both hydrophobically and ionically with

432

compounds during NC assembly.

433

ACS Paragon Plus Environment

22

Page 23 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Biomacromolecules

434

ASSOCIATED CONTENT

435

Supporting Information. Additional information on particle characterization is available free of

436

charge via the Internet at http://pubs.acs.org.

437

AUTHOR INFORMATION

438 439 440 441 442 443 444

Nikolas t. Weissmueller† Hoang D. Lu† Amanda Hurley Robert K. Prud’homme

445

Corresponding Author

446

*E-mail: [email protected]

447

Acknowledgements

448

The authors would like to thank Dr. Christina Tang for assistance with TEM imaging. We would

449

like to thank Professor Martin Semmelhack for CAI-1 material, and Professor Bonnie Bassler for

450

materials used in cell work.



Co-first authors

451 452

Funding

453

Funds were provided by Princeton University’s internal SEAS Old Guard grant, Princeton

454

University Center for Health and Wellbeing, and the Woodrow Wilson School of Public and

455

International Affairs Program in Science, Technology, and Environmental Policy.

456 457 458

ACS Paragon Plus Environment

23

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 24 of 29

459 460

References:

461 462 463 464 465 466 467 468 469 470 471 472 473 474 475 476 477 478 479 480 481 482 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501

1. Prabhu, R. H.; Patravale, V. B.; Joshi, M. D., Polymeric nanoparticles for targeted treatment in oncology: current insights. International journal of nanomedicine 2015, 10, 100118. 2. Wang, A. Z.; Langer, R.; Farokhzad, O. C., Nanoparticle delivery of cancer drugs. Annual review of medicine 2012, 63, 185-98. 3. Kumari, A.; Yadav, S. K.; Yadav, S. C., Biodegradable polymeric nanoparticles based drug delivery systems. Colloids and surfaces. B, Biointerfaces 2010, 75 (1), 1-18. 4. Hudson, D.; Margaritis, A., Biopolymer nanoparticle production for controlled release of biopharmaceuticals. Critical reviews in biotechnology 2014, 34 (2), 161-79. 5. Matteucci, M. E.; Brettmann, B. K.; Rogers, T. L.; Elder, E. J.; Williams, R. O.; Johnston, K. P., Design of potent amorphous drug nanoparticles for rapid generation of highly supersaturated media. Mol Pharmaceut 2007, 4 (5), 782-793. 6. Matteucci, M. E.; Miller, M. A.; Williams, R. O.; Johnston, K. P., Highly Supersaturated Solutions of Amorphous Drugs Approaching Predictions from Configurational Thermodynamic Properties. Journal of Physical Chemistry B 2008, 112 (51), 16675-16681. 7. Matteucci, M. E.; Paguio, J. C.; Miller, M. A.; Williams, R. O.; Johnston, K. P., Flocculated amorphous nanoparticles for highly supersaturated solutions. Pharmaceutical Research 2008, 25 (11), 2477-2487. 8. Matteucci, M. E.; Paguio, J. C.; Miller, M. A.; Williams, R. O.; Johnston, K. P., Highly Supersaturated Solutions from Dissolution of Amorphous Itraconazole Microparticles at pH 6.8. Mol Pharmaceut 2009, 6 (2), 375-385. 9. Bagga, S.; Adams, H. P.; Rodriguez, F. D.; Kemp, J. D.; Sengupta-Gopalan, C., Coexpression of the maize delta-zein and beta-zein genes results in stable accumulation of deltazein in endoplasmic reticulum-derived protein bodies formed by beta-zein. The Plant cell 1997, 9 (9), 1683-96. 10. Wang, S. Z.; Esen, A., Primary structure of a proline-rich zein and its cDNA. Plant physiology 1986, 81 (1), 70-4. 11. Argos, P.; Pedersen, K.; Marks, M. D.; Larkins, B. A., A structural model for maize zein proteins. The Journal of biological chemistry 1982, 257 (17), 9984-90. 12. Momany, F. A.; Sessa, D. J.; Lawton, J. W.; Selling, G. W.; Hamaker, S. A. H.; Willett, J. L., Structural characterization of alpha-zein. J Agr Food Chem 2006, 54 (2), 543-547. 13. Matsushima, N.; Danno, G.; Takezawa, H.; Izumi, Y., Three-dimensional structure of maize alpha-zein proteins studied by small-angle X-ray scattering. Bba-Protein Struct M 1997, 1339 (1), 14-22. 14. Wu, Y.; Holding, D. R.; Messing, J., Gamma-zeins are essential for endosperm modification in quality protein maize. Proceedings of the National Academy of Sciences of the United States of America 2010, 107 (29), 12810-5. 15. Wang, Y.; Padua, G. W., Nanoscale Characterization of Zein Self-Assembly. Langmuir 2012, 28 (5), 2429-2435. 16. Wang, Q.; Xian, W.; Li, S.; Liu, C.; Padua, G. W., Topography and biocompatibility of patterned hydrophobic/hydrophilic zein layers. Acta biomaterialia 2008, 4 (4), 844-51.

ACS Paragon Plus Environment

24

Page 25 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

502 503 504 505 506 507 508 509 510 511 512 513 514 515 516 517 518 519 520 521 522 523 524 525 526 527 528 529 530 531 532 533 534 535 536 537 538 539 540 541 542 543 544 545 546 547

Biomacromolecules

17. Podaralla, S.; Perumal, O., Influence of formulation factors on the preparation of zein nanoparticles. AAPS PharmSciTech 2012, 13 (3), 919-27. 18. Wang, H. J.; Lin, Z. X.; Liu, X. M.; Sheng, S. Y.; Wang, J. Y., Heparin-loaded zein microsphere film and hemocompatibility. Journal of controlled release : official journal of the Controlled Release Society 2005, 105 (1-2), 120-31. 19. Hurtado-Lopez, P.; Murdan, S., Formulation and characterisation of zein microspheres as delivery vehicles. J Drug Deliv Sci Tec 2005, 15 (4), 267-272. 20. Zou, T.; Gu, L. W., TPGS Emulsified Zein Nanoparticles Enhanced Oral Bioavailability of Daidzin: In Vitro Characteristics and In Vivo Performance. Mol Pharmaceut 2013, 10 (5), 2062-2070. 21. Lee, S.; Alwahab, N. S.; Moazzam, Z. M., Zein-based oral drug delivery system targeting activated macrophages. International journal of pharmaceutics 2013, 454 (1), 388-93. 22. Luo, Y.; Teng, Z.; Wang, Q., Development of zein nanoparticles coated with carboxymethyl chitosan for encapsulation and controlled release of vitamin D3. J Agric Food Chem 2012, 60 (3), 836-43. 23. Regier, M. C.; Taylor, J. D.; Borcyk, T.; Yang, Y. Q.; Pannier, A. K., Fabrication and characterization of DNA-loaded zein nanospheres. J Nanobiotechnol 2012, 10. 24. Lai, L. F.; Guo, H. X., Preparation of new 5-fluorouracil-loaded zein nanoparticles for liver targeting. International journal of pharmaceutics 2011, 404 (1-2), 317-23. 25. Muthuselvi, L.; Dhathathreyan, A., Simple coacervates of zein to encapsulate Gitoxin. Colloid Surface B 2006, 51 (1), 39-43. 26. Patel, A.; Hu, Y. C.; Tiwari, J. K.; Velikov, K. P., Synthesis and characterisation of zeincurcumin colloidal particles. Soft Matter 2010, 6 (24), 6192-6199. 27. Luo, Y. C.; Zhang, B. C.; Whent, M.; Yu, L. L.; Wang, Q., Preparation and characterization of zein/chitosan complex for encapsulation of alpha-tocopherol, and its in vitro controlled release study. Colloid Surface B 2011, 85 (2), 145-152. 28. Wu, Y. P.; Luo, Y. G.; Wang, Q., Antioxidant and antimicrobial properties of essential oils encapsulated in zein nanoparticles prepared by liquid-liquid dispersion method. Lwt-Food Sci Technol 2012, 48 (2), 283-290. 29. Parris, N.; Cooke, P. H.; Hicks, K. B., Encapsulation of essential oils in zein nanospherical particles. J Agr Food Chem 2005, 53 (12), 4788-4792. 30. Xiao, D.; Davidson, P. M.; Zhong, Q. X., Spray-Dried Zein Capsules with Coencapsulated Nisin and Thymol as Antimicrobial Delivery System for Enhanced Antilisterial Properties. J Agr Food Chem 2011, 59 (13), 7393-7404. 31. Hu, K.; McClements, D. J., Fabrication of surfactant-stabilized zein nanoparticles: A pH modulated antisolvent precipitation method. Food Res Int 2014, 64, 329-335. 32. Zhang, Y.; Niu, Y.; Luo, Y.; Ge, M.; Yang, T.; Yu, L. L.; Wang, Q., Fabrication, characterization and antimicrobial activities of thymol-loaded zein nanoparticles stabilized by sodium caseinate-chitosan hydrochloride double layers. Food chemistry 2014, 142, 269-75. 33. Chen, H. Q.; Zhong, Q. X., A novel method of preparing stable zein nanoparticle dispersions for encapsulation of peppermint oil. Food Hydrocolloid 2015, 43, 593-602. 34. Li, K. K.; Zhang, X.; Huang, Q.; Yin, S. W.; Yang, X. Q.; Wen, Q. B.; Tang, C. H.; Lai, F. R., Continuous preparation of zein colloidal particles by Flash NanoPrecipitation (FNP). J Food Eng 2014, 127, 103-110. 35. Patel, A. R.; Bouwens, E. C. M.; Velikov, K. P., Sodium Caseinate Stabilized Zein Colloidal Particles. J Agr Food Chem 2010, 58 (23), 12497-12503.

ACS Paragon Plus Environment

25

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

548 549 550 551 552 553 554 555 556 557 558 559 560 561 562 563 564 565 566 567 568 569 570 571 572 573 574 575 576 577 578 579 580 581 582 583 584 585 586 587 588 589 590 591 592

Page 26 of 29

36. Wang, L. J.; Yin, Y. C.; Yin, S. W.; Yang, X. Q.; Shi, W. J.; Tang, C. H.; Wang, J. M., Development of Novel Zein-Sodium Caseinate Nanoparticle (ZP)-Stabilized Emulsion Films for Improved Water Barrier Properties via Emulsion/Solvent Evaporation. J Agr Food Chem 2013, 61 (46), 11089-11097. 37. Podaralla, S.; Perumal, O., Preparation of zein nanoparticles by pH controlled nanoprecipitation. Journal of biomedical nanotechnology 2010, 6 (4), 312-7. 38. Dickinson, E., Properties of emulsions stabilized with milk proteins: Overview of some recent developments. J Dairy Sci 1997, 80 (10), 2607-2619. 39. Johnson, B. K.; Prud'homme, R. K., Flash NanoPrecipitation of organic actives and block copolymers using a confined impinging jets mixer. Australian Journal of Chemistry 2003, 56 (10), 1021-1024. 40. Johnson, B. K.; Prud'homme, R. K., Chemical processing and micromixing in confined impinging jets. AIChE Journal 2003, 49 (9), 2264-2282. 41. Liu, Y.; Cheng, C. Y.; Liu, Y.; Prud'homme, R. K.; Fox, R. O., Mixing in a multi-inlet vortex mixer (MIVM) for flash nano-precipitation. Chem Eng Sci 2008, 63 (11), 2829-2842. 42. D'Addio, S. M.; Baldassano, S.; Shi, L.; Cheung, L. L.; Adamson, D. H.; Bruzek, M.; Anthony, J. E.; Laskin, D. L.; Sinko, P. J.; Prud'homme, R. K., Optimization of cell receptorspecific targeting through multivalent surface decoration of polymeric nanocarriers. Journal of Controlled Release 2013, 168 (1), 41-49. 43. Gindy, M. E.; DiFelice, K.; Kumar, V.; Prud'homme, R. K.; Celano, R.; Haas, R. M.; Smith, J. S.; Boardman, D., Mechanism of macromolecular structure evolution in self-assembled lipid nanoparticles for siRNA delivery. Langmuir 2014, 30 (16), 4613-22. 44. Pinkerton, N. M.; Gindy, M. E.; Calero-Ddel, C. V.; Wolfson, T.; Pagels, R. F.; Adler, D.; Gao, D.; Li, S.; Wang, R.; Zevon, M.; Yao, N.; Pacheco, C.; Therien, M. J.; Rinaldi, C.; Sinko, P. J.; Prud'homme, R. K., Single-Step Assembly of Multimodal Imaging Nanocarriers: MRI and Long-Wavelength Fluorescence Imaging. Advanced healthcare materials 2015, 24;4(9):1376-85. 45. Pinkerton, N. M.; Grandeury, A.; Fisch, A.; Brozio, J.; Riebesehl, B. U.; Prud'homme, R. K., Formation of stable nanocarriers by in situ ion pairing during block-copolymer-directed rapid precipitation. Mol Pharm 2013, 10 (1), 319-28. 46. Gindy, M. E.; Prud'homme, R. K., Multifunctional nanoparticles for imaging, delivery and targeting in cancer therapy. Expert opinion on drug delivery 2009, 6 (8), 865-78. 47. D'Addio, S. M.; Baldassano, S.; Shi, L.; Cheung, L.; Adamson, D. H.; Bruzek, M.; Anthony, J. E.; Laskin, D. L.; Sinko, P. J.; Prud'homme, R. K., Optimization of cell receptorspecific targeting through multivalent surface decoration of polymeric nanocarriers. Journal of controlled release : official journal of the Controlled Release Society 2013, 168 (1), 41-9. 48. Tang, C.; Amin, D.; Messersmith, P. B.; Anthony, J. E.; Prud'homme, R. K., Polymer directed self-assembly of pH-responsive antioxidant nanoparticles. Langmuir 2015, 31 (12), 3612-20. 49. Lu, H. D.; Spiegel, A. C.; Hurley, A.; Perez, L. J.; Maisel, K.; Ensign, L. M.; Hanes, J.; Bassler, B. L.; Semmelhack, M. F.; Prud'homme, R. K., Modulating Vibrio cholerae quorumsensing-controlled communication using autoinducer-loaded nanoparticles. Nano Lett 2015, 15 (4), 2235-41. 50. Dickey, L. C.; McAloon, A.; Craig, J. C.; Parris, N., Estimating the cost of extracting cereal protein with ethanol. Industrial Crops and Products 1999, 10 (2), 137-143.

ACS Paragon Plus Environment

26

Page 27 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

593 594 595 596 597 598 599 600 601 602 603 604 605 606 607 608 609 610 611 612 613 614 615 616 617 618 619 620 621 622 623 624

Biomacromolecules

51. Higgins, D. A.; Pomianek, M. E.; Kraml, C. M.; Taylor, R. K.; Semmelhack, M. F.; Bassler, B. L., The major Vibrio cholerae autoinducer and its role in virulence factor production. Nature 2007, 450 (7171), 883-6. 52. van Henegouwen, G. M. B.; Junginger, H. E.; de Vries, H., Hydrolysis of RRR-αtocopheryl acetate (vitamin E acetate) in the skin and its UV protecting activity (an in vivo study with the rat). Journal of Photochemistry and Photobiology B: Biology 1995, 29 (1), 45-51. 53. Khdour, O. M.; Lu, J.; Hecht, S. M., An acetate prodrug of a pyridinol-based vitamin E analogue. Pharmaceutical research 2011, 28 (11), 2896-2909. 54. Figueroa, C. E.; Reider, P.; Burckel, P.; Pinkerton, A. A.; Prud'homme, R. K., Highly loaded nanoparticulate formulation of progesterone for emergency traumatic brain injury treatment. Therapeutic Delivery 2012, 3 (11), 1269-1279. 55. Lu, H. D.; Spiegel, A. C.; Hurley, A.; Perez, L. J.; Maisel, K.; Ensign, L. M.; Hanes, J.; Bassler, B. L.; Semmelhack, M. F.; Prud’homme, R. K., Modulating Vibrio cholerae QuorumSensing-Controlled Communication Using Autoinducer-Loaded Nanoparticles. Nano letters 2015, 15 (4), 2235-2241. 56. Luo, Y.; Teng, Z.; Wang, T. T.; Wang, Q., Cellular uptake and transport of zein nanoparticles: effects of sodium caseinate. J Agric Food Chem 2013, 61 (31), 7621-9. 57. Kumar, V.; Wang, L.; Riebe, M.; Tung, H. H.; Prud'homme, R. K., Formulation and Stability of ltraconazole and Odanacatib Nanoparticles: Governing Physical Parameters. Mol Pharmaceut 2009, 6 (4), 1118-1124. 58. Liu, Y.; Kathan, K.; Saad, W.; Prud'homme, R. K., Ostwald ripening of beta-carotene nanoparticles. Physical Review Letters 2007, 98 (3), 036102. 59. Ogawa, K., Effects of salt on intermolecular polyelectrolyte complexes formation between cationic microgel and polyanion. Advances in colloid and interface science 2015, 226, 115-121. 60. Solomatin, S. V.; Bronich, T. K.; Eisenberg, A.; Kabanov, V. A.; Kabanov, A. V., Colloidal stability of aqueous dispersions of block ionomer complexes: effects of temperature and salt. Langmuir 2004, 20 (6), 2066-2068. 61. Pansare, V. J.; Bruzek, M. J.; Adamson, D. H.; Anthony, J.; Prud'homme, R. K., Composite Fluorescent Nanoparticles for Biomedical Imaging. Molecular Imaging and Biology 2014, 16 (2), 180-188.

625 626 627 628 629 630 631

TOC graphic summary:

ACS Paragon Plus Environment

27

Biomacromolecules

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 28 of 29

632 Mixer Geometry

V. cholerae (inactive)

Organic 100% EtOH

Core (VitE-Ac) Drug (Dye, CAI-1) Hydroalcoholic 60% EtOH

Zein

CAI-1 in NP (Aqueous)

Aqueous

Free CAI-1 (DMSO)

Buffer: 10mM Sodium citrate, 150µM citric acid, pH 7.5 Aqueous

MIVM FNP

V. cholerae (active)

Casein in buffer

633

ACS Paragon Plus Environment

28

Page 29 of 29

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Biomacromolecules

Mixer Geometry

Organic 100% EtOH

Core (VitE-Ac) Drug (Dye, CAI-1) Hydroalcoholic 60% EtOH

Zein Aqueous

Buffer: 10mM Sodium citrate, 150μM citric acid, pH 7.5 Aqueous

MIVM FNP

Casein in buffer

ACS Paragon Plus Environment