CFAD V. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 1 of 16
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 1 of 16
`
`

`
`BIOLOGICAL TRACE ELEMENT RESEARCH:
`
`Published under the auspices ofthe International Association ofBioinorganic Scientists
`
`Oflicial Journal ofthe World Biomedical Selenium Society
`Gerhard N. Schrauzer - Editor-in-Chief
`Biological Trace Element Research Institute - San Diego, CA
`Manuel F. Flores-Arce - Managing Editor
`Laboratory of Trace Elements and Environmental Toxicology, Instituto Tecnologico de Tijuana, Mexico
`Charles H. Hill - Editor - College of Agriculture and Life Sciences - North Carolina State University -
`Raleigh - NC 27695
`Peter Schramel - Editor, Trace Element Analysis - Institut fur Okologische Chemie - GtS, [ngolstadter
`Lanstrasse #1 - D-8402 Neuherberg, Germany
`Yasushi Kodama - Editor - School of Medicine - University of Environmental and Occupational Health -
`Iseigaoka Yahata Nishi Ku - Kitakyushu, Japan 807
`Alain Favier - Editor - Laboratoire de Biochemie C, CHU de Grenoble, Hopital A. Michallon, BP 217,
`38043 Grenobole, France
`
`T. Akiyama, University of Occupational
`and Environmental Health, Kitakyushu, Japan
`J. A. Centeno, AFIP, Washington, DC
`G. F. Combs, Jr., Comell University, Ithaca, NY
`M. Costa, New York University Medical Center, NY, NY
`R. J. Cousins, University of Florida, Gainesville, FL
`U. M. Cowgill, Carbondale, CO
`A. T. Diplock, Guy's Hospital Medical School, London, UK
`José L. Domingo, Universitat Rovirai Virgili, Reus, Spain
`' I. E. Dreosti, CSIRO, Adelaide, Australia
`' L. Flohé, GBF, Braunschweig, Germany
`A. Furst, University of San Francisco, San Francisco, CA
`E. Harper, University of California, San Diego, La Jolla, CA
`. R. I. Henkin, Georgetown University Hospital,
`Washington, DC
`V. Iyengar, NIST, Gaithersburg, MD
`T. Jukes, University of California, Berkeley, CA
`M. Krachler, Research Center of Jiilich, Jiilich, Gennany
`P. Laszlo, University de Liege, Liege, Belgium
`B. Ldnnerdal, University of California, Davis, CA
`C. J. Lovatt, University of California. Riverside, CA
`
`Editorial Board
`E. J. Massaro, Duke University, Durham, NC
`D. M. Medeiros, Ohio State University, Columbus, OH
`F. H. Nielsen, USDA ARS, Grand Forks, ND
`R. Osterberg, Swedish University of Agricultural
`Sciences, Uppsala, Sweden
`W. Pories, East Carolina School of Medicine,
`Greenville, NC
`P. D. Saltman, University of California, La lolla, CA
`H. H. Sandstead, University of Texas, Galveston, TX
`B. Sarkar, Hospital for Sick Children. Toronto, Canada
`J. E. Spallholz, Texas Tech University, Lubbock, TX
`F. W. Sunderman, Jr., University of Connecticut
`Medical School, Fannington, CT
`G-q. Wang, Xi'an Medical University,
`Xi'an Shaanxi, PROC
`P. D. Whanger, Oregon State Univ., Corvallis, OR
`D. R. Williams, University of Wales, Cardiff, Wales, UK
`G. Yamada, Kumamoto University, Kumamoto, Japan
`P. F. Zatta, CNR Center, University of Padova, Italy
`A. Zgirsky, University of Lodz. Poland
`
`Volume 66, Numbers 1-3, Winter 1998
`
`Copyright © l998 Humana Press Inc. All Rights Reserved.
`
`This publication is printed on acid-free paper.
`ANSI Z39.48-1984 (American National Standards Institute) Permanence of Paper for Printed Library Materials,
`
`No part of this publication may be reproduced or transmitted in any form or by any means, electronic or
`mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in
`writing from the copyright owner. All authored papers, editorials, news, comments, opinions, conclusions, or
`recommendations are those of the author(s), and do not necessarily reflect the views of the publisher.
`
`Biological Trace Element Research is made available for abstracting or indexing in Chemical Abstracts,
`Current Contents, Index Medicus. Excerpta Medica, Biological Abstracts, Science Citation Index, and
`appropriate related compendia.
`
`Subscription Information: Biological Trace Element Research (ISSN 0163-4984) is published l8 times per year
`(monthly + two extra volumes) in six volumes. Subscription price for Vols. 6 l—66 is $775 (US) and $825 (foreign); please add
`$40 (US) or $60 (foreign) for postage.
`POSTMASTER: Send address changes to Biological Trace Element Research, Humana Press, 999 Riverview Dr., Suite
`208, Totowa, NJ 075 12. All correspondence, subscription orders, and notices ofchange ofaddress (both old and new addresses
`must be provided) should be sent at least 8 weeks in advance ofthe date ofeffect to Humana Press. Periodicals postage is paid
`at Passaic, NJ, and at additional mailing offices, by the Humana Press.
`Photocopy Authorization Policy: Authorization to photocopy items for internal or personal use, or the internal or personal
`use of specific clients, is granted by Humana Press, provided that the base fee ofUS $8.00 per copy, plus US $00.25 per page,
`- is paid directly to the Copyright Clearance Center Inc, 222 Rosewood Drive, Danvers, MA 01923. For those organizations that
`ave been granted a photocopy license from the CCC, a separate system ofpayment has been arranged and is acceptable to
`ana Press. The fee code forpusers of the Transactional Reporting Service is: 0163-4984/98 $8.00 + $00.25.
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 2 of 16
`
`

`
`Biological Trace Element Research
`
`Vol. 66: Nos. 1-3: Winter 1998
`
`Proceedings of the Second International Symposium
`
`I
`
`on the Health Efiects of
`
`Boron
`
`and Its Compounds
`
`October 22-24, 1997
`
`GUEST EDITORS
`
`B. DWIGHT CULVER
`
`FRANK M. SULLIVAN
`
`F. JAY MURRAY
`
`JAMES R. COUGHLIN
`
`PHILIP L. STRONG
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 3 of 16
`
`

`
`Biological Trace Element Research
`
`Vol. 66, Nos. 1-3, Winter 1998
`Cowmvrs
`
`Second International Symposium on the Health Effects of Boron and Its Compounds:
`Introduction
`
`B. Dwight Culver ................................................................................................................ 1
`Considerations in the Determination of Boron at Low Concentrations
`R. ‘Gregory Downing,* Philip L. Strong, B. Michael Hovanec, and
`lack Northington ................................................................................................................ 3
`
`A Round—Robin Determination of Boron in Botanical and Biological Samples
`R. Gregory Downing* and Philip L. Strong ...................................................................... 23
`
`Isotope Ratio Determination in Boron Analysis
`Ram N. Sah* and Patrick H. Brown ...................................................................... .. 39
`
`Measurement of Borate in Occupational Environments
`Robert A. Smith* and Frederick M. Ascherl ...................................................................... 55
`
`A Comparison of Worker Exposure to Inhalable and Total Dust, Inorganic Arsenic,
`and Borates Using Two Types of Particulate Sampling Assemblies in a Borate
`Mining and Processing Facility
`M. A. Katchen,* V. A. Puhalovich, R. Swaroop, and B. D. Culver .............................. 59
`
`Dietary Boron Intakes of Selected Populations in the United States
`Susan L. Meacham* and Curtiss D. Hunt .......................................................................... 65
`
`Multicountry Estimation of Dietary Boron Intake
`Charlene Rainey* and Leslie Nyquist .............................................................................. .. 79
`
`-Sources of Human Exposure: Overview of Water Supplies as Sources of Boron
`[ames R. Coughlin ................................................................................................................. .. 87
`
`In Vivo Percutaneous Absorbtion of Boron as Boric Acid, Borax, and Disodium
`Octaborate Tetrahydrate in Humans: A Summary
`Ronald C. Wester,* Xiaoying Hui, Howard 1. Maibach, Kathleen Bell,
`Michael]. Schell, D. [ack Northington, Philip L. Strong,
`and B. Dwight Culver ................................................................................................... .. 101
`
`In Vitro Percutaneous Absorbtion of Boron as Boric Acid, Borax, and Disodium
`Octaborate Tetrahydrate in Human Skin: A Summary
`Ronald C. Wester,* Tracy Hartway, Howard I. Maibach, Michael I. Schell,
`D. Iack Northington, B. Dwight Culver, and Philip L. Strong ........................... .. 111
`
`Boron Exposure from Consumer Products
`Margaret Richold ......................
`................................................................... .. 121
`Distribution of Boron in the Environment
`Peter Argust ........................................................................................................................... 131
`
`The Isotopic Composition of Anthropogenic Boron and Its Potential Impact
`on the Environment
`
`Avner Vengosh ........................................................................
`
`............................................ 145
`
`A Review of Boron Effects in the Environment
`............. 153
`Paul D. Howe ..
`.........................................................................
`_j__
`
`‘For papers with multiple authorship,the asterisk identifies the author to whom correspondence
`and reprint requests should be addressed.
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 4 of 16
`
`

`
`Biological Trace Element Research
`
`Vol. 66, Nos. 1-3, Winter 1998
`
`CONTENTS
`
`Relationships Between Boron Concentrations and Trout in the Firehole River,
`Wyoming: Historical Information and Preliminary Results ofa Field Study
`Ioseph S. Meyer/* Ann M. Boelter, Daniel F. Woodward, Iack N. Goldstein,
`Aida M. Farag, and Wayne A. Hubert ..........................................................._._............. 167
`
`Review of the Scientific Basis for Establishing the Essentiality of Trace Elements
`Walter Mertz .......................................................................................................................... 185
`
`Determining Human Dietary Requirements for Boron
`Barbara Sutherland/* Phil Strong, and Ianet C. King ..................................................193
`
`Regulation of Enzymatic Activity: One Possible Role of Dietary Boron in Higher
`Animals and Humans
`
`Curtiss D. Hunt ................................................................................................................... ..205
`
`The Nutritional and Metabolic Effects of Boron in Humans and Animals
`8. Samrnan,* M. R. Naghii, P. M. Lyons Wall, and A. P. Verus ............................... .. 227
`
`Adverse Reproductive and Developmental Effects in Xenopus from
`Insufficient Boron
`
`Douglas I. I-‘ort,* Timothy L. Propst, Enos L. Stover, Philip L. Strong,
`and P. Iay Murray .......................................................................................................... .. 237
`
`The Response of Trout and Zebrafish Embryos to Low and High Boron
`Concentrations Is U—Shaped
`Ruby 1. Rowe, Collen Bouzan, Sam Nahili, and Curtis D. Eckhert* ......................... 261
`
`Assessing the Effects of Low Boron Diets on Embryonic and Fetal Development
`in Rodents Using In Vitro_ancl In Vivo Model Systems
`Louise Lanoue, Marie W. Taubeneck, Iesus Muniz, Lynn A. Hanna,
`Philip L. Strong, F. Iay Murray, Forrest H. Nielsen, Curtiss D. Hunt,
`and Carl L. Keen* .............................................................................................................271
`
`The Importance of Boron Nutrition for Brain and Psychological Function
`Iames G. Penland ................................................................................................................ .. 299
`
`The Justification for Providing Dietary Guidance for the Nutritional Intake of Boron
`Forrest H. Nielsen ............................................................................................................... .. 319
`
`A Comparative Review of the Pharmacokinetics of Boric Acid in Rodents
`and Humans
`
`F. Iay Murray ....................................................................................................................... .. 331
`
`Comparative Toxicology of Borates
`Susan A. Hubbard ................................................................................................................. 343
`
`Developmental Effects of Boric Acid in Rats Related to Maternal Blood Boron
`Concentrations
`
`Catherine I. Price,‘ Philip L. Strong,
`F. Iay Murray, and Margaret M. Goldberg .............................................................. .. 359
`
`
`*Forpapers with multiple authorship,the asterisk identifies the author to whom correspondence
`and reprint requests should be addressed.
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 5 of 16
`
`

`
`Biological Trace Element Research
`
`Vol. 66: Nos. 1-3, Winter 1998
`Cozvrenrs
`
`Effects of Boric Acid on Axial Skeletal Development in Rats
`Michael G. Narotsky,* Iudith E. Schmid, Iames E. Andrews,
`and Robert I. Kavlock ..
`..................................................... 373
`
`The Effects of Dietary Boric Acid on Bone Strength in Rats
`Robert E. Chapin,* Warren W. Ku, Mary Alice Kenney, and Harriet McCoy ....... .. 395
`
`Comparison of Infertility Rates in Communities from Boron-Rich and
`Boron-Poor Territories
`E. Tficcar, A. H. Elhan,* Y. Yavuz, and B. S. Sayli ..................................................... .. 401
`
`An Assessment of Fertility in Boron-Exposed Turkish Subpopulations:
`2. Evidence That Boron Has No Effect on Human Reproduction
`Bekir Sitki Sayli ................................................................................ ..
`
`The Effect of Essentiality on Risk Assessment
`George C. Becking .................................................................................... ..
`
`International Programme on Chemical Safety (IPCS) Environmental Health Criteria
`on Boron Human Health Risk Assessment
`
`George C. Becking* and Bing-Heng Chen ...................................................................... .. 439
`
`Boron Tolerable Intake: Re—evaluation of Toxicokineticsfor Data—DeriUed
`Uncertainty Factors
`Michael Dourson,* Andrew Maier, Bette Meek, Andrew Renwick,
`Edward Ohanian, and Kenneth Poirier ......................................................................453
`
`_
`Summary of Research Needs
`Frank M. Sullivan and B. Dwight Culver*....~........... .;................................................... .. 465
`
`-
`
`Author Index..................................................................................................................... ..469
`
`Subject Index..................................................................................................................... ..471
`
`
`
`*For papers with multiple authorship,the asterisk identifies the author to whom correspondence
`and reprint requests should be addressed.
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 6 of 16
`
`

`
`©Copyright 1998 by Humana Press Inc.
`All rights of any nature, whatsoever, reserved.
`0l63—4984/98/G601-3-0001 $08.25
`
`Second International Symposium
`on the Health Effects of Boron
`
`and Its Compounds
`
`Introduction
`
`The Second International Symposium on the Health Effects of Boron
`and Its Compounds, October 22—24, 1997, brought together investigators
`interested in the effects of boron in biological and environmental systems
`at toxicological, pharmacological, and nutritional levels. A majority of
`the principal investigators working in these areas reported the results
`of the studies they have conducted over the past five years. Important
`contributions were made also by participants interested in the research
`methods required to measure these effects, especially at low—exposure
`levels where the complex chemistry of boron must be taken into account.
`Studies pointing to the nutritional importance of boron have been
`conducted" over the past 20 years, and at the 1992 "symposium, a number
`of studies investigating the nutritional role of boron were presented.
`However, it was not until this present symposium that the essentiality of
`boron for some vertebrate species was demonstrated.
`As was the case with the First International Symposium held in
`1992, it was hoped that this symposium would, by presenting as com-
`pletely as possible the current state of knowledge about boron and its
`compounds, stimulate research into issues identified by the participants.
`The concluding article in this volume is an effort to underline future
`research needs.
`
`Biological Trace Element Research
`
`Vol. 66, I998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 7 of 16
`
`

`
`Coughlin
`
`Guidelines for Cam-
`ml Health Canada,
`
`\ ublished data cited
`Document: Boron,
`
`and R. G. Downing,
`~environmenta1 and
`
`©Copyright 1998 by Humana Press Inc.
`All rights of any nature, whatsoever, reserved.
`0163-4984/98/6601-3-0101 $10.25
`
`In Vivo Percutaneous Absorption
`of Boron as Boric Acid, Borax,
`and Disodium Octaborate
`
`Tetrahydrate in Humans
`
`~d
`‘ ation of
`elféartloln in the
`
`A Sunlmary
`
`RONALD C. WESTER,*" XIAOYING HuI,1
`HOWARD I. MAIBACH,‘ KATHLEEN BELL,2
`MICHAEL J. ScHELI_,3 D. JACK NoRTHII~IoToI~I,“
`PHILIP STROl‘lG,5 AND B. DWIGHT CLILVER6
`
`’Department of Dermatology, University of California, San
`Francisco, CA; 2Department of Nutrition, University of California,
`San Francisco, CA; ‘Department of Biostatistics, University
`of North Carolina, Charlotte, NC; ‘West Coast Analytical Service,
`Santa Fe Springs, CA; 5U.S. Borax Inc., Valencia, CA;
`and 6Department of Medicine, University of California, Irvine, CA
`
`ABSTRACT
`
`Literature from the first half of this century reports concern for
`toxicity from topical use of boric acid, but assessment of percutaneous
`absorption has been impaired by lack of analytical sensitivity. Analyti-
`cal methods in this study included inductively coupled plasma-mass
`spectrometry, which now allows quantitation of percutaneous absorp-
`tion of 10B in 1°B~enriched boric acid, borax, and disodium octaborate
`tetrahydrate (DOT) in biological matrices. This made it possible, in the
`presence of comparatively large natural dietary boron intakes for the
`in vivo segment of this study, to quantify the boron passing through
`skin. Human volunteers were closed with 10B-enriched boric acid,
`5.0%, borax, 5.0%, or disodium octaborate tetrahydrate, 10% in aque-
`ous solutions. Urinalysis, for boron and changes in boron isotope
`ratios, was used to measure absorption.
`Boric acid in Vivo percutaneous absorption was 0.226 (SD = 0.125)
`mean percent dose, with flux and permeability constant (KP) calculated
`
`*Author to whom all correspondence and reprint requests should be addressed.
`
`Biological Trace Element Research
`
`101
`
`Vol 66, 1998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 8 of 16
`
`

`
`Wester et al.
`
`at 0.009 ug/crn2/h and 1.9 X 10"7 cm/h, respectively. Borax absorption
`was 0.210 (SD = 0.194) mean percent dose, with flux and KP calculated
`at 0.009 ug/cm2/h and 1.8 X 10-7 cm/h, respectively. DOT absorption
`was 0.122 (SD = 0.108) mean percent, with flux and KP calculated at
`0.01 ug/cm?/h and 1.0 X 10'7 cm/h, respectively. Pretreatment with
`the potential skin irritant 2% sodium lauryl sulfate had no" effect on
`boron skin absorption.
`These in vivo results show that percutaneous absorption of
`boron, as boric acid, borax, and disodium octaborate tetrahydrate,
`through intact human skin is low and is significantly less than the
`average daily dietary intake. This very low boron skin absorption
`makes it apparent that, for the borates tested, the use of gloves to pre-
`vent systemic uptake is unnecessary. These findings do not apply to
`abraded or otherwise damaged skin.
`
`Index Entries: Boron analysis; isotope ratio; excretion; dietary
`boron.
`
`INTRODUCTION
`
`Boron (B) is an ubiquitous element in rocks, soil, and water. Most of
`the earth's soils have <10 ppm B, with high concentrations found in parts
`of the western US and in other sites stretching from the Mediterranean
`to Kazakhstan. The average soil B concentration is 10-20 ppm, and sea-
`water contains an average of 4.6 ppm B. Fresh waters normally range
`from <0.01 to‘1.5 ppm, with higher concentrations in regions of high B
`soil levels. Being present in every part of the environment, it is likely that
`life evolved in the presence of the element. B is essential to plant life.
`Borates are used widely in industrial and domestic applications. The
`manufacture of glass and related uses consume about half of the borates
`as boric anhydride (B203) in the US. Fiberglass accounts for the largest
`share, comprising about 77% of the total (54% for insulation fiberglass,
`23% for textile grade). Heat-resistant pyrex and other low—thermal-
`expansion glasses consume about 18%, whereas enamels, frits, and
`ceramic glazes consume 5%. Small amounts are used in sealing and opti-
`cal glasses, Vycor, and vitrifying nuclear waste (1).
`Thus, it is obvious that human skin comes into Contact with B min-
`erals and chemicals from mining to manufacturing to consumer use.
`However, this ubiquitous element is also present in all plant life and,
`thus, a part of everyone’s diet. This presented a classical situation of dis-
`tinguishing between a potentially small amount of B chemicals absorb-
`ing through skin against a much larger background of B in the daily diet.
`Classically, the way to distinguish the two has been to use a radiotracer
`for the skin-absorbing chemical. For B this is not chemically possible to
`do. Fortunately, new analytical technology "is now available, and this
`made it possible to distinguish between low levels of skin~absorbing B
`chemicals against a larger dietary background.
`
`Biological Trace Element Research
`
`Vol. 66, 1998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 9 of 16
`
`

`
`. Wester et al.
`
`Percutaneous Absorption of Boron
`
`MATERIALS AND METHODS
`
`Test articles, 10B-enriched boric acid, 1°B—enriched borax, and di-
`sodium octaborate tetrahydrate (DOT) were provided by U.S. Borax Inc.
`(Valencia, CA). Sodium lauryl sulfate (SLS) was obtained from Sigma
`Chemical Co. The dosing formulations, 5% boric acid solution and 5%
`borax solution, were prepared by adding the appropriate volume of
`water to the appropriate weighed amount of test compound. Samples
`were sonicated to ensure homogeneity; 10% disodium octaborate tetrahy-
`drate solution was formulated by U.S. Borax Inc. Stability of the formu-
`lations was shown for the course of the study.
`The in vivo dose was 1.8—mL solutions over 900-cm? skin area, or
`2 ul/cmz, the maximum dose that would not run off the skin. Human
`volunteers were recruited from the University of California, San Fran-
`cisco (UCSF) and the surrounding San Francisco Bay Area community.
`Twenty—four normal, healthy, males or females, aged 22-50 yr, were
`selected for this study. Before the study, all volunteers were instructed on
`how to keep a daily food diary and on what food items high in B to
`avoid during the course of the study
`The studies were divided into three groups and each group contained
`eight volunteers. Background urine was collected for the first 4 (1. Groups
`I, II, and III received two separate topical applications of 10B-enriched 5%
`boric acid, 5% borax, or 10% DOT solutions on their back skin, respec-
`tively. One dose was _applied_on day 5 under normal skin conditions, and -
`the other on day 12 under potentially irritated skin conditions created by
`applying 2% SLS solution. Twenty—four hours after each topical dose,
`residual chemical on the closed skin site was removed by skin wash.
`Twenty—four-hour urine samples were collected daily for 17 d. Urine
`samples from day 1 to day 4 were used to establish base B levels and iso-
`tope ratios in the urine. The samples from day 5 to days 11 and 12 to the
`end were used to compare absorbed level under normal skin and irri-
`tated skin conditions. To evaluate the dosing site skin condition, the
`transepidermal water loss TEWL measurement and skin visual scoring
`were taken each time before dosing (including SLS treatment) and wash-
`ing. Five sites of the marked area, the upper-left corner, the upper—right
`corner, the center, the lower-left corner, and the lower-right corner, were
`used for the TEWL measurement.
`
`On day 5, the physical condition of the back skin of each volunteer
`was examined, and then a 900-cmz area, 30 X 30 cm, was marked on the
`back. The marked area received a single topical application of 5% boric
`acid (group I), 5% borax (group H), or 10% DOT (group III). Each dose for-
`mulation was delivered with a 3.0—mL Eppendorf polypropylene syringe
`(Brinkmann Instru_ments, Ind., Westbury, NY). The delivered dose was
`quantitated by weighing the micro syringe before and after dosing.
`After topical application, the dosed area was allowed to air-dry, and
`then the volunteer dressed in a commercial white T-shirt for 24 h. The vol-
`
`Biological Trace Element Research
`
`Vol. 66, 1998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 10 of 16
`
`

`
`Wester et al.
`
`unteers were requested not to touch or wash the dosed area for 24 h.
`Thus, the dosing site
`tected for the 24-h dosing period. Twenty-
`four hours after dosing
`'
`y removed, and the
`dosed site was washed using gauze p
`’
`'
`MO), IvoryTM liquid soap (Proctor 8: G
`The washing procedure was as follows:
`1. 50% Ivory soap solution (V/V).
`2. Distilled deionized water.
`3. 50% Ivory soap solution (v/v).
`IL-10. Distilled deionized water.
`
`time of day as the first
`SLS solution on the
`
`On day 11, the vol
`time of dosing and was
`same 900-cm? area of the back. On day 1
`gle topical application of 5% boric acid (group I), 5% borax (group II), or
`DOT (group III) was applied as on day 5. After topical application, the
`dosed area was allowed to air-dry, and then the volunteer dressed in a
`commercial white T-shirt for 24 h. Again volunteers were requested not
`to touch or wash the dosed area for 24 h.
`Twenty-four hours after the second closing (day 13), the T-shirt was
`carefully removed, and the closed site was washed using gauze pads
`1), Ivory liquid soap (Proctor 8: Gamble), and water.
`_
`_
`sured by the
`Courage 8: Khazaka Electronic
`GmbH (Cologne, Germany). Five sites on the treated skin were used for
`each measurement. Those were the upper-left corner, the upper—right cor-
`ner, the center, the lower-left corner, and lower-right corner. Each mea-
`surement was taken for at least 3 min. The average value of the last 20 s
`and the standard deviations were recorded when the measurement value
`was stable. Prior to the measurement, the subject rested in the test room
`at least 30 min; room temperature and humidity were recorded. During
`the study (on days 5, 6, 11, 12, an
`the volunteers’ skin was checked
`by a trained dermatologist for erythema and edema (redness and inflam-
`mation). Skin condition was scored on a scale of 0-5.
`In order to avoid excess dietary B intake, the volunteers were asked
`to follow some dietary restrictions during the 17-d study. Foods and bev-
`erages to avoid were dried fruits (primes, raisins, dates, and so forth),
`includin foods containin these items; cherries, avocados, nuts
`eanuts,
`S
`8
`P
`pecans, almonds, and so on), peanut butter, honey, prune juice/grape
`juice, wine, and beer. In addition, the volunteers were advised to check
`labels and avoid products containing boric acid or sodium borate, such
`as health food supplements, pharmaceuticals, skin ointments, cosmetics,
`and mouthwash or dental solutions. The volunteers were also asked to
`and beverages they con- :
`keep a detailed food diary of all of the foods
`In each daily record, the ,
`sinned for each of the 17 d during the study.
`
`;'
`.
`
`Biological Trace Element Research
`
`Vol. 66, 1998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 11 of 16
`
`

`
`1, St. Louis,
`I and Water.
`
`Percutaneous Absorption of Boron
`
`105
`
`Volunteer showed the time food/beverage was consumed, the location
`where it was consumed,
`type of food or beverages, method of food
`preparation, all
`ingredients, estimated amount eaten, and standard
`household measurements or the weight in ounces or grams.
`Samples were analyzed for 10B and “B content by inductively coupled
`plasma mass spectrometry (ICPIVIS) using a VG PlasmaQuad II (VG Ele-
`mental, Danvers, MA). Samples were delivered by peristaltic pump (Gilson
`Minipuls 3, Middleton, WI) at approx 1.2 mL/min. SRM 951, boric acid
`from the National Institute of Standards and Technology (NTST, Gaithers—
`burg, MD), which is certified for both purity and isotopic content, was used
`to prepare B standards in the range of 10—1000 pg/L. The acid concentra-
`tion in the standards was matched to the sample for accurate quantitation.
`
`RESULTS
`
`Figures 1, 2, and 3 show the in Vivo daily plot of excess 10B as indi-
`vidual and mean values for boric acid, borax, and DOT, respectively. The
`figures illustrate the study design for the 17—d sample collection period,
`with 10B-enriched dosing occurring on days 5 (nontreated skin) and 12
`(pretreatment with 2°/o SLS).
`Table 1 shows that 0.226 (SD = 0.125) mean percent dose of 5% boric
`acid was absorbed through the skin in human volunteers over a 6-d
`period after a 24-h dosing period. Pretreatment with 2% SLS for 24 h gave
`nochange in‘ mean percent dose absorbed (mean = 0.239, SD = 0.174).
`With 5% borax, the mean percent dose absorbed was 0.210 (SD : 0.194)
`with no treatment, and no change was seen with 2% SLS pretreatment
`(mean = 0.184, SD = 0.219). With 10% DOT,
`the mean percent dose
`absorbed was 0.122 (SD = 0.108) with no treatment, and no change was
`noted with 2% SLS pretreatment (mean = 0.107, SD = 0.133). No statisti-
`cally significant difference was found in the percent B absorption of the
`three compounds, either without pretreatment (p = 0.33) or after 2% SLS
`pretreatment (p = 0.35), even though the mean percent B absorption of
`DOT was about half of that obtained for both boric acid and borax. No
`skin irritation was noted for any of the above treatments.
`Table 2 gives the recovery and accountability of administered dose
`in 24-h urine accumulation (percent dose absorbed), the T-shirt that was
`worn over the dosed skin for 24 h, and the washed skin after the 24-h
`dosing period. The T-shirts were able to contain dose within contact of
`the skin for the 24~h period. Also, skin wash at 24 h removed chemical
`from the skin. Thus, it can be assumed that administered dose was in
`contact with the skin of the Volunteers for the 244h closing period. Also,
`skin wash number 10 (last wash) contains a <1% dose; thus, the skin
`washing procedure seems complete. It is assumed that the remainder of
`the dose was lost to outside clothing (over the T-shirt) and bedding
`(sheets, blankets) during the 24-h dosing period.
`
`Biological Trace Element Research
`
`VOL 65. 1993
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 12 of 16
`
`

`
`Wester et al.
`
`0 lndividualvalue
`—O~ Mean value
`
`
`
`ExcessBoron10We)
`
`10
`
`12
`
`Day
`
`Fig. 1. Daily plot of excess B 10 for in Vivo group topically dosed with
`[NB]-boric acid on days 5 and 12. Individual and mean values are shown.
`
`0.30
`
`'
`
`0 Individual value
`~O— Mean value
`
`|..°0UI
`
`A $VOY
`
`‘coI
`
`-0
`
`III
`til
`cn
`oo
`]
`
`xH
`
`12
`
`14
`
`16
`
`18
`
`Fig. 2. Daily plot of excess B 10 for" in vivo group topically dosed with
`[NB]-borax on days 5 and 12. Individual and mean values are shown.
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 13 of 16
`
`

`
`Waste,» et a1_
`
`Percutaneous Absorption of Boron
`
`O lndividualvalue
`-0- Mean value
`
`0.18
`
`0.16
`
`0.14
`
`0.12
`
`0.10
`
`0.08
`
`0.06
`
`0.04
`
`0.02
`
`0.00
`
`-0.02
`
`-0.04
`
`-0.06
`
`
`
`ExcessBoron10(‘’/o)
`
`Fig. 3. Daily plot of excess B 10 for in Vivo group topically dosed with
`[1°B]-DOT on days 5 and 12. Individual and mean values are shown.
`
`Table 1
`
`In Vivo Percutaneous Absorption of 10B as 5% Boric Acid, 5% Borax, and 10%
`DOT in Normal Human Volunteers
`Mean Percent Dose Absorbed‘
`
`Compound
`
`51 Boric Acid
`51 Borax
`10% DOT
`
`No Pre~Treat:ment
`so
`95% CI”
`.125
`(.121,
`.331)
`.194
`(.048.
`.372)
`.108
`(.032,
`.212)
`
`Mean
`.225
`.210
`.122
`
`'
`
`’
`
`.
`
`.
`
`2% SLS Pre—Trear.menI:
`so
`95% cf’
`.174
`(.094,
`.384)
`.219
`(.001,
`.367)
`.133
`(.004,
`.219)
`
`“fl : 8; 24-h dosing period.
`”95% confidence interval for the mean based on the t distribution.
`CPaired t—test p-value for the difference in mean percent dose absorbed.
`
`Biological Trace Element Research
`
`Vol. 66,
`
`I 998
`
`CFAD v. Anacor, IPR2015-01776, CFAD EXHIBIT 1061 - Page 14 of 16
`
`

`
`108
`
`Wester et al.
`
`Table 2
`Dose Accountability for 10B as 5°/o Boric Acid, 5% Borax, and 10% DOT
`in Normal Human Volunteers
`
`Parameter
`
`Percent Dose Mean (S.D.)
`St Boric Acid
`5! Borax
`
`( n = 8)
`
`Percent Dose Absorbed
`No treatment
`SL5 treatment
`T-shirt
`(24 hours)
`No treatment
`SL5 treatment
`Skin Wash (No.1-9)
`No treatment
`SLS treatment
`Skin Hash (No.
`No treatment
`SL5 treatment
`
`0.226 (0.125)
`0.239 (0.174)
`
`0.210 (0.194)
`0.185 (0.219)
`
`50.9 (16.6)
`53.3 (5-9)
`
`V
`
`6.1 (1.8)
`5.3 (2.3)
`
`0.4 (0.40)
`0.3 (0.13)
`
`47.4 (12.4)
`51-2 (4-0)
`
`10.6 (1.8)
`6.9 (4.1)
`
`0.9 (0.25)
`.=——
`0.7 (0.26)
`
`DISCUSSION
`The undertaking of this study presented a challenge not frequently
`encountered by those who attempt to measure percutaneous absorption
`of chemica