US008277762B2 (12) United States Patent (10) Patent No.: US 8,277,762 B2 Newsam et al. (45) Date of Patent: Oct. 2, 2012 (54) APPARATUS AND METHODS FOR (56) References Cited EVALUATING THE BARRIER PROPERTIES OF A MEMBRANE U.S. PATENT DOCUMENTS (75) Inventors: John M. Newsam, San Diego, CA (US); 4493,815. A * 1/1985 Fernwood et al. ............ 422,101 Ilya Feygin, Mountainside, NJ (US); (Continued) Samir Mitragotri, Goleta, CA (US); Robert Dominic King-Smith, San FOREIGN PATENT DOCUMENTS Diego, CA (US) GB 2246081 A 1, 1992 (73) Assignee: Tioga Research, Inc., San Diego, CA (Continued) (US) OTHER PUBLICATIONS (*) Notice: Subject to any disclaimer, the term of this Supplementary Partial European Search Report dated Aug. 17, 2007 patent is extended or adjusted under 35 issued in respect of corresponding Application No. EP 0477 9728. U.S.C. 154(b) by 1298 days. Supplementary European Search Report dated Jun. 11, 2008 issued in respect of corresponding Application No. EP 0477 9728. (21) Appl. No.: 10/566,648 Examiner's Report dated Sep. 2, 2010 issued in respect of corre sponding Application No. CA 2,534,359. (22) PCT Fled: Jul. 30, 2004 Primary Examiner — Jill Warden (86) PCT NO.: Assistant Examiner — Dwayne K Handy S371 (c)(1), (74) Attorney, Agent, or Firm — Foley & Lardner LLP (2), (4) Date: Aug. 16, 2006 (57) ABSTRACT (87) PCT Pub. No.: WO2OOS/O12549 The present invention teaches apparatus (511) and methods PCT Pub. Date: Feb. 10, 2005 for screening the effect of test formulations on the barrier (65) Prior Publication Data properties of a membrane (212), that are especially beneficial when skin is used as the test membrane (212). The apparatus US 2007/O183936A1 Aug. 9, 2007 (511) and methods enable more efficient measurements of skin permeabilization, of the permeation of molecular or par Related U.S. Application Data ticulate entities through skin, and of the absorption and (60) Provisional application No. 60/491,553, filed on Aug. adsorption by skin of ingredients in fluid formulations, 1, 2003. together with screening of exfoliation of material from the exterior of the stratum corneum. The apparatus (511) provide (51) Int. C. for fluid contact to the skin from both donor and receptor BOIL 3/00 (2006.01) sides, for measurements of skin electrical response in the (52) U.S. C. ........ 422/552; 422/547; 422/548; 422/551; presence of test formulations, of permeation and permeation 422/553; 422/559; 436/178 enhancement, for the depth profiling of test formulation con (58) Field of Classification Search ............... 422/82.01, stituents through the skin, of stratum corneum component 422/83, 102,547,548,551, 552, 553,559, disruption, and of loss of material from the stratum corneum. 422/560, 561; 436/149, 174, 181, 178 See application file for complete search history. 27 Claims, 14 Drawing Sheets )ZZZZZZZiz22.ÉZ 2NNN, N NSNS/ arear 7 raca 7-a-sers ZY US 8,277,762 B2 Page 2 U.S. PATENT DOCUMENTS 7.494,622 B2 * 2/2009 Picollet-Dahan et al. 422/82.01 5,141,719 A 8, 1992 Fernwood et al. 2001/0042710 A1 11/2001 Clarket al. 5,342.581 A 8, 1994 Sanadi 2002/0006643 A1* 1/2002 Kayyem et al. .............. 435,912 5,888,830 A * 3, 1999 Mohan et al. ................. 436,174 2005/02601 01 A1* 11/2005 Nauck et al. .................... 422.99 6,045,755 A * 4, 2000 Leblet al. ....................... 506,33 6,338,802 B1 1, 2002 Bodner et al. FOREIGN PATENT DOCUMENTS 6,455,007 B1 9, 2002 Mansky et al. WO 9839099 A1 9, 1998 6,485,690 B1* 11, 2002 Pfostet al. .................... 422/552 WO OO25922 A2 5, 2000 6,742,659 B2 6, 2004 Clark et al. WO 03008112 A1 1, 2003 6,805,842 B1* 10, 2004 Bodner et al. ................ 422/102 6,817,558 B1 11, 2004 Karlsson et al. * cited by examiner U.S. Patent Oct. 2, 2012 Sheet 1 of 14 US 8,277,762 B2 12 Design Experimentation Campaign Implement Screening protocol Define Performance Provide membrane, test formulations Targets Complete experimentation stages Analyze results 11 Performance Targets Achieved? Proceed to Scale Up, Commercialization 15 FIG. 1 U.S. Patent Oct. 2, 2012 Sheet 2 of 14 US 8,277,762 B2 213 FIG. 2A FIG. 2B U.S. Patent Oct. 2, 2012 Sheet 4 of 14 US 8,277,762 B2 FIG. 4 U.S. Patent Oct. 2, 2012 Sheet 5 of 14 US 8,277,762 B2 Ø Z ---------- %)!!!!!!!!!!24ZijØ2,22,225,225,2||2||2 FIG. 5B F.G. SA U.S. Patent Oct. 2, 2012 Sheet 6 of 14 US 8,277,762 B2 614 615 613 611 612 617 616 621 620 624 618 619 622 623 FIG. 6 U.S. Patent Oct. 2, 2012 Sheet 7 of 14 US 8,277,762 B2 712 711 712 Sy 713 Nayaaaaaaaayaaaaaaaaaaaaaa. 714. FIG. 7A 711 712 715 711 720 N4 NA. NAN 716 717 718 719 FIG. 7B FIG. 7C FIG. 7 U.S. Patent US 8,277,762 B2 814 22222222 815 817 1. FIG. 8B [] 2 2 2 813 812 U.S. Patent Oct. 2, 2012 Sheet 9 of 14 US 8,277,762 B2 FIG. 9A FIG. 9B FIG. 9C FIG. 9D FIG. 9E E - E - E - - - - E - E SEE FIG. 9G FIG. 9H FIG. 9 U.S. Patent Oct. 2, 2012 Sheet 10 of 14 US 8,277,762 B2 1011 1012 1017 FIG. 10A 1018 1019 1013 1014 1016 1015 1011 1012 1017 FIG. 10B 1020 1021 1015 1011 1012 1022 1023 FIG. 10C 1014 1015 FIG. 10 U.S. Patent Oct. 2, 2012 Sheet 11 of 14 US 8,277,762 B2 FIG, 11A 118 FIG. 11 U.S. Patent Oct. 2, 2012 Sheet 12 of 14 US 8,277,762 B2 1211 1211 1212 1218 al 2.a. a. 1213 1217 FIG. 12 U.S. Patent Oct. 2, 2012 Sheet 13 of 14 US 8,277,762 B2 Determine lamina details, Sourcing, treatment protocols 1311 Determine membrane barrier screening system (MBSS) protocol 1313 1312 Determine test formulation SCOpe, Sampling & 1314 sequence (library set") Provide test formulation library 1317 Library Set Apply measurement ASSessment complete? protocol in MBSS Perform Off-line analyses 1316 Analyze and interpret Cumulative data 1318 FIG. 13 U.S. Patent Oct. 2, 2012 Sheet 14 of 14 US 8,277,762 B2 Provide donor array plate Introduce samples into donor array Wells Mix samples in donor Wells Apply lamina to donor plate Apply receptor array Fill receptor array Well(s) SeCure MBSS assembly Contact donor Well samples to lamina Iteratively poll electrode array Sample & analyze receptor wells Disassemble MBSS Analyze final donor & receptor fluids; depth profile lamina FIG, 14 US 8,277,762 B2 1. 2 APPARATUS AND METHODS FOR route of administration as (i) it avoids first-pass liver metabo EVALUATING THE BARRIER PROPERTIES lism of the drug, (ii) circumvents exposure of the drug to the OF A MEMBRANE chemical rigors of the gastrointestinal (“GI) tract, (iii) may permit delivery of drugs with short biological half-lives and/ CROSS-REFERENCE TO RELATED or narrow therapeutic windows, (iv) may reduce adverse APPLICATIONS events in patients such as GI distress, (v) may offer more uniform plasma dosing of the drug, (vi) allows prompt inter This application claims the benefit of Provisional Applica ruption of dosing, and (vi) may increase patient compliance. tion No. 60/491,553, filed on Aug. 1, 2003, which is incor Active transdermal delivery routes, in which an external porated herein by reference. 10 stimulus is applied to drive the drug through the barrier, include iontophoresis, Sonophoresis, electroporation, FIELD OF THE INVENTION microneedles, and application of high Velocity solid particles The present invention relates generally to the development 1 or liquids 2.3 (see 4). These all require the application of methods for modifying the barrier properties of a mem 15 of a physical device, with irritation and compliance often an issue. A passive delivery route, in which a formulation con brane. Such as skin, and to rapid and efficient methods of taining the drug needs simply to be applied to the skin is measuring the effect of chemicals on the barrier properties. Substantially preferred. The prime requirements, in general, BACKGROUND OF THE INVENTION for a passive transdermal route are (I) that Sufficient skin permeation can be achieved, (ii) that skin irritation and skin Membranes, both natural and synthetic, play critical roles sensitization be avoided, and (ii) that reasonable delivery in many fields. The utility of Such membranes depends on a efficiency be accomplished. number of factors, physical, mechanical, chemical and bio Only a small number of drugs have been approved for logical. In many cases, of central issues are the barrier prop application in transdermal patches (including Scopolamine, erties of the membrane, which determine the degree to and 25 nitroglycerin, clonidine, estradiol, nicotine, fentanyl, test rate at which species of various types can pass through the osterone, norelgestromin with ethinyl estradiol). These share membrane. It is often desirable to modify these barrier prop the three characteristics of (I) low molecular mass (<500 Da) erties for the specific use. One of the most evident of natural 5. (ii) high lipophilicity, and (iii) Small required dose (up to membranes is skin. milligrams) 4. Skin as a Barrier 30 Passive approaches for transdermal delivery of drugs based The skin, the largest organ of the human body, has three on vesicles, such as liposomes, have shown some promise for layers the epidermis, dermis and subcutis. The subcutis, the other classes of molecules 6-9. However, these technologies deepest layer, provides thermal insulation and has a shock have yet to appear in an FDA-approved transdermal patch absorbing effect that helps protect the body's organs from product despite more than 20 years of work on vesicle-based injury.