`
`MAY 1966
`vol. 50; 110- 4
`
`U_S_ DEPARTMFNT OF HEALTH EDUCATION, AND WELFARE
`
`,
`
`'
`
`Public Healtl
`
`1 S
`
`°
`
`ervlce
`
`'
`
`|nnoPharma Exhibit 10620001
`
`

`

`CANCER CHEMOTHERAPY Reports
`NATIONAL CANCER INSTITUTE
`
`Kenneth M. Endicott, Director
`
`SCIENTIFIC EDITORS
`
`Jerome B. Block, John P. Glynn, and J. A. R. Mead
`
`EDITORIAL STAFF
`
`Evelyn E. Parker, Managing Editor
`Patricia Morrison, Assistant Managing Editor
`Bethany Viera, Assistant Editor
`
`
`EDITORIAL POLICY
`
`Original contributions are welcome on all aspects of the treatment of cancer. Pre-
`liminary reports of work in progress are acceptable to facilitate the prompt exchange
`of
`information among investigators in this field. Appropriate commentaries and
`reviews are invited. Letters pertinent
`to published articles may also be submitted.
`Requests for the publication of symposia will be considered provided a complete edited
`transcript of the proceedings is submitted.
`
`Manuscripts should be addressed to:
`
`Managing Editor
`Cancer Chemotherapy Reports
`National Cancer Institute
`Bethesda, Maryland 20014
`
`INSTRUCTIONS TO AUTHORS
`
`Summary
`Begin the article with a short summary.
`Text
`
`Submit an original and 2 carbon copies of the manuscript, typed double spaced. Each page Sh0111d begin With
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`Tables
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`References
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`In book references, give author,
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`in text should begin with number 1 and continue consecutively.
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`Illustrations
`Photographs and graphs preferably should be clear, glossy prints. Original art work should be done in black ink
`for good reproduction.
`Nomenclature
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`Names of compounds should conform to American usage. See Chemical Abstracts for proper nomenclature.
`Abbreviations
`Please note that periods have been eliminated from all abbreviations of units of measurement and from abbre-
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`Th is material was empied
`at the NLM and may Ina
`Su bjeezt Ufitlopsg-‘right: Laws
`
`|nnoPharma Exhibit 1062.0002
`
`

`

`FEQUANTITATIVE COMPARISON or TOXICITY or ANTICANCER AGENTS
`IN MOUSE, RAT, HAMSTER, DOG, MONKEY, AND MAN 3-;
`Emil J:.Freireich,3 Edmund A. lGehan,‘ David P.1Rc1l|,5 Leon H.'Schmidf," and Howard E.'Skippei-7
`
`SUMMARY
`
`(1)
`
`large
`Toxicity data from small animals (mouse, rat, and hamster),
`animals (dog and monkey), and humans were gathered, placed on a rea-
`sonably similar basis, and compared quantitatively. Each animal species
`and all species combined were used to predict the toxic doses in man (based
`on mg/In“ of surface area). Two models were assumed for the relationship
`between the maximum tolerated dose (MTD) in man and the approximate I
`LD10 in each animal system:
`(dose in man) = (dose in animal system 1')
`
`and
`(2)
`, 6)
`.
`.
`.
`(dose in man) = A; X (dose in animal system i), (i = 1 ,
`where A. is the fraction of the dose in animals used to predict the dose in
`humans (assumed different for each animal system, ie, 73 = 1 ,
`.
`.
`.
`, 6). It
`was found that when animal systems other than the rat were used the very
`simple model (1) was remarkably good for predicting the MTD in humans,
`though model (2) leads to slightly better predictions. Based on model (2),
`the animal systems are ranked in order of predictive ability: rhesus mon-
`key, Swiss mouse, rat, BDF, mouse, dog, and hamster. The best estimate
`of the MTD in man is made by weighting the estimates from the various
`animal species. Dose on an mg/m‘ basis is approximately related to dose
`on an mg/kg basis by the formula
`. ,7)
`.
`.
`(«i = 1 ,
`(dose in mg/m’) = (km). >< (dose in mg/kg),
`where (lam). is the appropriate factor for converting doses from mg/kg to
`mg/m” surface area for each species. When the (lc'm)i factors are known,
`equally good predictions of MTD in man can be made by either dose unit.
`on an mg/m” basis, the MTD in man is about the same as that in each
`animal species. On an mg/kg basis, the MTD in man is about ‘/12 the LD10
`in mice, 1/J the LD10 in hamsters, 1/, the LD10 in rats, M, the MTD in
`rhesus monkeys, and 1/2 the MTD in dogs. In each case the ratio is the
`(km) factor in the animal system to that in man. Hence relationships
`among the Various animal species and man are somewhat simpler and
`more direct on an mg/m” basis. These results support the conclusion that
`the experimental test systems used to evaluate the toxicities of potential
`anticancer drugs correlate remarkably closely with the results in man.
`
`,
`
`(
`f
`I
`Q
`
`L
`
`I I
`
`'
`‘
`
`I
`
`
`
`.—~—..———-v——~.——
`
`1""‘,“~
`iwved Dec 29, was revised Jan
`c;..‘;2:’°i:;:°:;yp:€.uE“a;al
`"' St d
`d
`under the auspices of the Acute L
`,
`_
`y
`7
`kemiauT§2,isl<0rI}‘(:)i'ce of the National Cancer Institute by
`J
`I “£36 _addreSs requests £01‘ feprlnts to Dr. Rall.
`the Subhuman Subcommittee.
`,
`Califggfigaait
`f°1‘ Prlmate B1010gY. UHW Of
`3 M. D, A i
`n Hospital, Houston, Tex.
`_
`_
`--
`4BiOmet1_;(e};,:nch’ Nationai Cancer
`I
`Inst, Public
`’Ket'teri_ng-Meyer Laboratory of Southern Research
`l Health Service, Bethesda, Md.
`Inst, Birmingham, Ala.
`I
`l
`CANCER CHEMOTHERAPY REPORTS VOL. 50, N0. 4, MAY 1966
`
`219
`
`l
`
`5 This material may be protected by Copyright law (Title 17 U.S. Code)
`. ,
`l
`
`I
`
`|nnoPharma Exhibit 10620003‘
`
`

`

`The biologic aspect of a
`drug development
`program to
`discover
`compounds
`effective ,-
`against any clinical disease is generally an ex-
`ercise in comparative pharamacology. In the
`typical program, compounds are screened in
`small animals against some easily produced an(l
`reproduced pathologic condition. A close rela-
`tionship must exist between the screening sys-
`tem and the ultimate clinical condition for the
`program to have the potential for success. Thus
`examination of this relationship is highly im-
`portant. In cancer chemotherapy the similari-
`ties and differences have often been considered
`among transplantable tumors, virus-induced
`tumors, carcinogen-induced tumors, and spon-
`taneous tumors iii animals, and between animal
`tumors and the various cancers and leukemias
`in man. However the similarities and differ-
`ences between mice, rats, hamsters, dogs, mon-
`keys, and man have been considered less often
`in terms of quantitative and qualitative aspects
`of the toxic effects of drugs. The consistency
`of the action of therapeutic agents among vari-
`ous mammalian species is a keystone of most
`drug development programs, yet only rarely has
`this been studied in a quantitative manner.
`Classically comparative pharmacology and
`physiology have been concerned with differences
`which permit analytic studies of specific bio-
`logic systems, and these studies have yielded
`valuable information. But it is equally impor-
`tant to consider the much more frequent simi-
`larities; we have tried to do this in the present
`analysis.
`
`Of all the toxicologic end points, lethal toxic-
`ity is the easiest to measure with reasonable
`precision. Therefore we considered the lethal
`dose of certain cancer chemotherapeutic agents
`in various laboratory animals. For man the
`end point was the maximum tolerated dose
`(MTD).
`llopefully two benefits might-accrue
`from this evaluation: (1) If there is reasonable
`consistency in the reactions of various mam-
`malian species,
`the toxicologic component Of
`cancer chemotherapy screening will be shown
`to have a rational basis. (2) If such consistency
`is found,
`the problems of introducing highly
`toxic therapeutic agents into man might be
`2_ppr()aChCd more confidently. If major incon-
`sistencies are discovered frequently, this would
`highlight the deficiencies in present screening
`systems and raise serious questions about the
`utility of these schemes for safe introduction of
`new drugs into man.
`220
`
`eutlc
`No attempt was made to relate tl1_1::‘3¥n
`doses in the various mammalian Sllecatgemp
`‘future this correlation should be hmt is no
`since the therapeutic target 1“ the
`(ml. if an
`the same as the toxicity targelfi H.°Wen experi-
`agent has therapeutic 1)1‘<>I>°rnCS In adose leV9
`mental system, it is well to know the stilication
`for Datients. Since there is some Juthesc dose
`for using‘ MTD’s in cancer tlierailyx
`levels were studied.
`s to
`The plan of this i~etrosp9°ljYe Stildigblzginetl
`examine considerable toxicol0£¥1c‘.da;:.y screen-
`in (a) small animals, used in .111 13:“g evalua-
`ing and quantitative sccondalyd monkeys, f01'
`tion; (I2) larger animals, 40955. an
`eggs of tox-
`the quantitative and qualitative asfnd (0) man,
`icity at sublethal and lethal levels. 3:0 determine
`the target species. The $0.31 was between cer-
`what relationship exiS’CS:_ If ally’
`d points in
`tain commonly used toxicologlc
`for it num-
`the various animal species and ind
`her of anticancer agents.
`t
`suagest
`Nothing‘ in this reP"”t is intel]1de(zlbl(t)3 immine-
`or imply that short cuts are .91 cl: dies. Dose-
`clinical or clinical
`toxicologlc f5‘1man arenot
`limiting and serious toxic el“fec’55 ln st carefully
`always apparent from even thc.IT1f3m_ma1S (1),
`done toxicologic investigations lllildarly wider,
`It is emphasized and should ()0 iflmpt to M.
`stood that it is dam/c*r'01.LS_t" (“,0 {city date
`tmpolatc dirvctl-y f7"’"‘ animal
`(lxNew drugs
`to maximum tolerated doses m ‘$11.7’-» 1 trial only
`can be introduced safely into cllflllfigrmacologic
`through careful toxicologlc and 1:1 tious study
`study in animals and then V91"yl°‘1ue1_ dosages
`in man, starting with much
`‘fw ted by the
`than those which appear to be '50 9”‘
`animals.
`5 STUDY
`APPROACHES AND ASSUMPTIONS IN THI
`hich
`.
`The published ami_unpubl1%h9d §g*§,,;;’,,,,e,,
`form the basis for this analy-$15 Ԥ.eIfe1,ent pm.
`by numerous investigators using hsistent and
`tocols and end points. We used Cothat the data
`.
`7
`were comparable. The biolosllc 9“
`l’
`reasonable general assumptions Sod Ointsy pI_0_
`-
`tions necesssafk
`tocols, assumptions, and correc
`,
`able are dc‘
`to make the results more compar
`Toxicolagic End Points (See Apflemllx
`scribed brieflY-
`_
`I)
`which
`d
`. _.
`Mouse, rat, or hamster: gtljnxill1?fg’u“:h:nd°S;:hedu1c
`when administered by
`C‘—/118,,’
`ie the LD10) dur-
`100 animals were used in il typical
`0
`km--* 3 S-1-e::‘:“.1:.:‘;:.e.':.‘;:r.:'.°.
`;,....;i.
`is so mg? the
`ing 3 §P0C1‘ ‘
`’
`,
`-
`‘
`d termina I011.’
`CANCER CHEMOTHERAPY REPORTS
`This material in
`at the N-LM and
`
`l
`
`-‘.~’.u“l:i~jae~:t Ufi.
`|nnoPharma Exhibit 1o62)ooo4
`
`

`

`1
`
`,
`
`,
`
`‘
`
`>
`
`typically 2-4 animals
`(IL) MTD;
`Dog or monkey:
`,
`i were used at each dose level, spaced by 2-fold incre-
`t merits.
`In all
`instances individual doses which killed
`0 and 100%; were used. The highest dose killing 0%
`‘, was considered the l\«ITD.
`(Ir) Dose-related, hema-
`tupoictic elfccts;
`localized hemorrhages of the gastro-
`intestinal tract; geiieralizerl hemorrhagic lesions (ab-
`doniinzil and thoracic viscera); stimulation of the cen-
`tral nervous system (CNS) ; others.
`Man:
`(ft) MTD for a fixed schedule (dose causing
`mild to moderate sublctlial toxic effects in a significant
`percent of patients);
`(b) MTD for a variable sched-
`ule, calculated from the daily dose and median period
`to toxic eilects i'e<1uii‘iiig cessation of drug; the judg-
`ment of many clinical
`investigators was necessarily
`accepted in making this estimation.
`Because of the nature of the available data,
`the toxicologic end points in the various ani-
`mal species were related to the MTD in man.
`‘ Although it was necessary to assume that the
`dosages resulted in the same percentage of tox-
`I
`icity in each species. the results do not depend,
`‘
`in a major way, on this assumption. For the
`drugs in this study,
`the dose-toxicity curves
`were relatively steep so that if the true per-
`centage of
`toxicity for a given ‘dosage was,
`say, between 5% and 15%,
`the actual dosage
`used would not differ veiy much from the dos-
`age that should have been used.
`It was necessary to use toxicologic data ob-
`tained by various routes of drug administra-
`tion, ie, intraperitoneal (ip) for small animals,
`oral for small animals and man, and intra-
`venous (iv) for large animals and man. In mice
`and rats the LD10’s obtained by the ip and 1V
`routes are usually comparable.
`Another variable for which some reasonable
`correction must be made is the dosage schedule
`including the total dose. We assumed that the
`toxicity of anticancer agents is cumulative.
`, G1-iswom at 31,
`(3)
`reported that when the
`LD10’s in BDF. mice of 70 agents, including
`the major classes of anticancer agents, were
`compared for two schedules, ‘Id 1"7 days and
`qd 1—11 days,“ the mean rati0_(<ld 1-7 d<1yS/
`qd 1—11 days) was 1.56. This is very close to
`that which might be expected from direct cumu-
`lative drug toxicity (11 days/7 days = 1-57)-
`Pinkel
`(2) and other investigators pointed
`out that the usual closes of certain drugs in
`various animal species and man were compara-
`ble when the dose was measured on the basis
`of mg/m“ of surface area. Consequently most
`of the results are presented in mg/In’. However
`since mg/kg is a commonly used unit of drug
`dosage, some results are also presented in this
`flqd = drug given once daily for as many days as
`indicated.
`
`4,
`,
`
`l
`i
`
`‘ VOL. 50, No. 4, MAY 1966
`was sea:-pi 5 cl
`‘ad may be
`’vp§n'ig‘h—t Laws
`
`.~
`,
`-
`‘t-
`.
`unit. Only a simple trangf
`to change me’/kg to mg/m??’€l.l§§?oi3§ iE2‘L‘éi‘Z‘i
`tionships developed are equivalent whichever
`unit
`is used. The quantitative relationships
`were simpler when expressed in mg/m,
`.
`A conversion factor (km) was used to trans.
`form mg/kg to mg/m’ by the equation mg/
`kg X (km) : mg/mi: (km) factors for ani-
`mals, given their weight, are presented in table
`1
`(Appendix II), and table 2 (Appendix II)
`Dresents a way of transforming doses in mg/
`>13: to mg/In’ for man, given height and body
`“’€i.‘lh‘E- Chsljt 1
`(Appendix II) is a diagram
`for detei*minrng surface area in man, given
`height and weight.
`Calculations based on units of body surface
`area have no intrinsic merit per se. Very likely
`some other basis such as surface area of the
`site of action of the drug, lean body mass, or
`some fractional power of body weight, D0531.
`bly related to length or some org'an-inembranc
`surface area, would be as appropriate or more
`appropriate. However the body surface area has
`been used to relate many physiologic pamm.
`eters among species and means of transform-
`ing the data are readily available. Further, in
`our clinical studies we routinely use body guy.
`ffilce a1"@afi0 adjust drug: dose for patients of
`different size and weight.
`RESULTS
`The first step in aiialyziiig the data was to
`correct the daily dosage schedules for man and
`for animals, when necessary,
`to a uniform
`schedule of qd 1-5 days. Thus if an LD10 for
`mice, or MTD for man, was obtained by a
`schedule of qd 1-10 days, we calculated that
`the LD10 (or MTD) for a schedule of (pi 1-5
`days was twice that value. The next step was to
`convert doses (LD10‘s or MTD’s) from mg/kg;
`to mg/ni’ This was accomplished by the ap-
`proximate formula
`, 7)
`.
`.
`(mg/kg), (i:'i, .
`><
`(mg/m‘) = (km).
`where the (lam). factor differs according to the
`species and also according to body Weight with.
`in each species.
`In the analysis an average
`(/WI): factor was used, assuming that individ-
`uals in each species were of average ]1eig]1L.t0_
`body-wei,crlit
`ratios, The (It-m)‘
`factors were
`derived from standard relationships between
`weight and surface area as given in Specter
`(40) and Seiidroy and Cecchini
`(39). Details
`and other inibrmation on relating drug doses
`in U12‘/kg to doses
`in mg/in’ are given in
`Appendix II.
`
`221
`
`|nnoPharma Exhibit 1062.0005
`
`

`

` EO
`0
`0
`CHART 2
`Comparison ul ioxicily cum on oniicaricfii
`
`ugenis ici inc mouse and nionloii a
`MG/M2 imsisi
`O Aiitiinaiaboiiies
`A Aikyiciimg agents
`0 Others
`
`
`
`
`
`
`5o
`
`.0
`
`o I
`
`_#J__h,4_i
`J
`I0
`‘O0
`'°°°
`i.o
`l
`aor Mousmowimc/Mzzooi-sooyschcdulel
`
`i000 i‘
`
`CHART 3
`
`Comparison at iaxicliy dam on uniicuncer
`uqeriis is! we iiuiiisiei cmi iiiuiiion a
`MG/M2 imsisi
`0 Aniimelahaiiies
`A Alkylalinq oqerils
`0 Oiheis
`
`
`O 0
`
`O
`
`.
`
`
`
`The basic data used in this study are given
`in table 1. Doses of 18 drugs" are presented
`in mg/kg and mg/m’ for the 6 species, along
`with source information and other pertinent
`data. An average dose (LD10 or MTD) of each
`drug was calculated from the multiple studies,
`if done, on each species. The average doses for
`the 6 animal systems and man are given in
`mg/kg in table 2, and in mg/m’ in table 3.
`Charts 1-6 indicate the closeness of the rela-
`tionship between the logarithm of the LD10, or
`MTD, in the various animal systems and in man
`when the dose is measured in mg/m’. Chart 7
`indicates the close relationship between 12
`times the LD10 in the BDF, mouse and the
`MTD in man when the dose is measured in
`mg/kg. The ratio of the (km) factors for an
`average man and a mouse is 37/3 = 12,3, It
`will be shown later that relationships between
`systems on an mg/kg basis are the same as
`those on an mg/m‘ basis if the ratio of (km)
`factors is considered.
`To examine further the relationship of dos-
`age, in mg/m’, between the animal systems and
`man, consider the following: For each animal
`system and man, there is a dose-toxicity curve.
`The basic data for each drug consist of esti-
`mates of a single point, the approximate LD10,
`on the dose-toxicity curves for man and the 6
`
`5cwo
`
`
`
`MAN=MAXlMUMTOLERATEDDCSE[MG/M2‘00I-5DeySchedule) 5
`
`0..
`
`I00
`I0
`ID
`SWISS M0USE'LD|0 (MG/M2‘ OD I-5 Day schedule]
`
`I000
`
`“Chemical Abstracts’ nomenclature‘ and NSC num-
`hers for the agents are given on page 243.
`222
`
`Th material we
`at. the i‘ii:i..‘i‘lifi‘ and
`
`:':»i.i5;i:ij«E=i:.“t, US Ci:i~p°g
`
`|nnoPharina Exhibit 10620006
`
` MANMAXIMUMTOLEHATEDDOSEIMG/M21ODI-5Dnyschedule) 55
`
`
`
`
`
`
`
`
`MAN.MAXiMUMTOLERATEDDOSEWG/MZIOUI-5Dayschedule)
`
` I 0.0
`
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`0.
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`I000
`
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`
`I00
`.09
`,0
`iiiiiiisiairmmiiiic/ii2=ooi-soayschedule}
`anima systems. We wish to describe the rela-
`tionship between the dose-toxicity curve for
`man and that for each of the anllnal S.VStemS~
`Two models are considered:
`(dose in man) : (dose in animal system i)
`(i=1,...,6)
`(1)
`and
`(dose in man) : A1 X (dose in animal syS<
`temi),
`(i=1,...,6).
`(2)
`Model
`(1)
`is a special case of mfldel
`(2)
`since they are the same when A. : 1. Model
`CANCER CHEMOTHERAPY REPORTS
`
`CHART i
`Cimipuiisan oi loiiiciiy data on uiiiicniicer
`ngeriis for the Swiss niousennd man
`[on u MG/M2 basis]
`
`
`0 Ilniimeiahcliles
`A Aikyiuiingogeiiis
`
`I Others
`
`
`O
`
`
`
`i0O
`
`5
`
`
`
`
`
`o
`
`

`

`cmwv 4 O
`
`
`Companson oihmcily dam on UYINCCHCEV
`0
`agents for the In! and munlnn cl MG/ A
`
`Nebusnsl
`0 Annmevubamss
`
`/\ Alkylcmig ugems
`%
`I Others
`
`
`
`
`5,Lu
`oc
`8i.4itLu_J
`
`5oo
`
`
`
`I00
`
`
`
`o_.
`
`
`
`(1(‘SGuyscnedulel
`
`F3
`
`
`
`IOUO
`
` ‘-00{-5Boysclieduiel 5o
`
`12Lu(/1oo
`
`
`oin
`(E_,
`o.-
`
`
`
`:0
`
`l
`
`O
`
`
`'0
`
` ’“Jim.....J
`.0
`woo
`V000
`RHESUS MONKEY:
`MAWUM TOLERIXTED DOSE(MG/Mz.QD |—5Dnyschedule)
`
`
`
`i______1,,jl
`I0
`:00
`i000
`1 0
`RAT: Lilm (M6/M100":-5 Day schedule]
`CHART 5
`Campcnsan of toxicity dam on unhcanccr
`
`ngenls tor ihe mesus mnnkey and man
`0
`
`(on u MG/M2 basis)
`0 Avmmevubclives
`A Alkylullng ugenls
`
`9 Ovhers
`
`5oo
`
`5o
`
`CHART G
`Cmnounson as Ioxxcwy dula um anticancer uqems my
`me dcqcnd monlon 0 MG/M2 basis]
`0 Annmevabomes
`A Alkylnlmq aqenls
`o cams
`
`0
`
`A
`
`
`
`
`MANIMAXIMUMTGLERATEDDOSEIMG/M2-QDIA5Dayschedule] 56 o
`
`
`
`MANMAXH/[UMIULERATEDDOSE[MG/KG¢0U|-DDaySchedule) 5 l
` 0Ol ————J
`
`loo —
`
`
`__l
`moo
`10
`0-I
`I0
`100
`DOGIMAXIMUM TOLERATED DOSE (MG/M2:0Dl—5 Day schedule)
`CHART 7
`Campanscn av Ioxncnydam on unlicuncev agents tor
`(he mouse and man (nnn MG/KG basis)
`The 12:1 velnhunsmp shown on a MG/KG bflsli KS eqmvuienl
`lo we H mammsmp shown onu MG/M2 basis (than 2)
`The nvproximule I21? Velfllldflihlp(lVlGU9€=|1lflll)lS In
`agreement Will!
`the who cl lhn KM factors used
`,0 _ for Mesa spe1:ies.ic,37=3[mun.muuse]=cuI2
`O Ammmubolnes
`A Alkylahnq uqems
`U Olhevs
`
`Q
`
`0
`
`O,| -
`
`am
`
`V0
`l0
`0.!
`am, MOUSE, Lo“, (Ma/moo l-5Day schedule]
`
`100
`
`(1) assumes that the dose in each animal sys-
`15 m gives a direct prediction of the dose in
`r:-an Model (2) assumes that the dose in man
`is a fraction (A.)
`01: the dos? "1 the ‘muilal
`gystem and the fraction remains constant oi
`the sample of dru2S>
`A third model was considered:
`'
`i
`= A X (dose in animal sys-
`(dmmmm)
`‘tam z)
`(i = 1,. .. ,6)
`Where B.
`is the power to which the C1080 is
`
`VOL, 50, NO. 4, MAY 1966
`I was amzeied
`rid m.a'e_,i baa
`}pwig:h-t La W5
`
`raised, assumed to be 1 in models (1) and (2).
`This model is a natural generalization of (2).
`However, since the estimates of Ba were near
`1
`for all animal systems,
`in fact within 1
`standard error (SE) limit, there is no advantage
`1.0 l1SiIl.‘I a more general model than (2).
`By these models, we wish to predict the dose
`in man from the dose in each animal system
`when both determinations are subject to samp-
`ling variation (‘and other assumptions as men-
`223
`
`|nnoPharma Exhibit 1062.000? ‘
`
`

`

`-
`Z
`0DSElVEElCl1dDYEdlClEll dosugeswo/M lmman
`using all unimalsyslamslweiqhled BSWll0iE5l
`o Model
`1
`0 Model 2
`°
`
`CHART 0
`
`U
`o
`
`C
`/9
`.
`0
`oo-
`O
`
`C
`K o
`
`moo
`AT I00
`2
`‘
`.a.7:1mo.=.
`E
`.2.
`'»‘—"Q.3u.II:n.
`
`242
`
`I300
`
`,
`
`0.:
`
`(:0
`I0
`I0
`'2
`MAMOBSEHVED DOSAGE [MG/M )
`on
`_
`.
`in table 5 and the weighted estimates bitidl in
`all animal systems combined are I310 an as
`chart 8. The best estimates of d,°5e 1-11 ‘$3171; 4,
`indicated by the standard dcv1é_1t}0“§ 1”‘ {mates
`are given by weighting the iI1d1V1d““ es ‘
`from each animal system.
`_
`_
`,
`Another model was considcfcd 1“ wlllcslesfiilrei
`dose in man (mg/m’) was related :90 , 0
`the animal species in a single equatlon‘
`
`tioned) in the sample of drugs. The statistical
`considerations in fitting these models are given
`in Appendix III.
`Model (1) is the simplest possible model; no
`parameters need to be estimated, Thus the
`doses in table 3 for each animal system are the
`predicted values of the dose in man and charts
`1-6 indicate that these predictions are reason-
`ably good. The standard deviations, on a log
`scale, of a predicted value of log (dose in man)
`were calculated for each animal system. The
`systems are ranked in order of predictive
`ability in the top half of table 4: monkey, Swiss
`mice, BDF. mouse, dog, rat, and hamster. A
`predicted value of the dose in man has been
`calculated by weighting the estimates from
`each animal system (see Appendix III) and
`the results are given in the last column of
`table 3. The standard deviation of a predicted
`value of log (dose in man) is 0.299, with multi-
`pliers of 0.50 and 2.0 for lower and upper
`standard deviation limits respectively. Thus the
`weighted estimate based on all systems is bet-
`ter than the estimate from any single system.
`Assuming model (2), the estimates of A. and
`Ar : 2 SE are given in the bottom half of table
`4. Note that the approximate 95% confidence
`limits for the multiplying factor, /1., include 1
`for all animals systems except the rat. Thus for
`the other animal systems it is reasonable to
`accept the very simple model (1) as providing
`an adequate prediction of the dose in man.
`However when all systems are combined to ob-
`tain an overall estimate of A; (see Appendix
`III), the approximate 95% confidence limits do
`not include 1. Also, note from the bottom half
`of table 4 that the standard deviation of a pre-
`dicted value of log (dose in man) is 0.275, al-
`most a 10% reduction from that of model (1).
`Therefore model
`(2)
`is preferred for fitting
`these data; however for future studies in which
`more precise estimates of LD10 are available,
`it may be that model (1) will be adequate.
`Using model (2), we can rank the animal
`systems in order of their predictive ability by
`considering the deviations of observed from
`predicted values of dose in man. These standard
`deviations are given in table 4. Thus the order
`is monkey, Swiss mouse, rat, BDF. mouse, dog,
`and hamster. The best predictions with model
`(2) are obtained by weighting the estimates of
`the dose in man from all 6 animal systems (the
`method is explained in Appendix III). The pre-
`dictions for the drugs in this study are given
`224
`
`log (dose in man) : 0.284 + 0347 10g (dose In
`Swiss IIIOUSC)
`V
`F
`A 1.061410% (dose In B17
`'
`mouse)
`,
`+ 0539 log (dose in rat)
`+ 0.801 log (dose 1“ m°“'
`l
`_
`10g (dose in dog‘)-
`
`lfn-.
`This predicting equation leads to a slight
`Drovement in the prediction of the (ins? Endnfim ’
`the deviations of observed f1‘9m_P1'ed1° 9 49 OS”
`ages were less (standard deviation Of 0-? fit 0&1
`log scale compared to 0.275 by “SW95 ‘I91? e '
`combined estimates). However £1
`l>Y‘€d1Ct_1°11 Of
`dosage in man cannot be made unless estimates
`of LD10 are available from all the animal sys-
`tems mentioned; also the model does not provide
`any real insight into the relationship between
`the dose-toxicity curve in each animal system
`and that in man.
`_
`_
`From considering charts 143, W15 q“‘3St101‘
`arose: Do the differences between the dose.
`
`CANCER CHEMOTHERAPY REPORTs
`ms. rrizatvzsrrial ii
`at it: E Fslihhvi am
`
`VE—lJ,'tl“_i-E‘;;t U53 {lite}:
`|nnoPharma Exhibit 1062.0008
`
`

`

`toxicity curves for man and for each animal
`system differ depending on whether an antime-
`tabolite or an alkylating agent was given‘! Us-
`ually the animal species, except the rat and
`monkey, underpredict the doses of antimetabo-
`lites and overpi-edict
`the doses of alkylating
`agents for man. By a statistical test (t test),
`there was some suggestion (P<0.10) that in
`Swiss mice and BDF, mice the predictions of
`dosage in man were lower for antimctabolites
`than for alkylating agents. There was no evi-
`dence of a difference in the other species. Only
`4 aiitimetabolites and 8 alkylating agents were
`tested in all animal species. Consequently fur-
`ther study is needed to determine whether the
`difference between dose—toxicity curves really
`depends on the type of agent.
`There is some value in comparing the rela-
`tionships found on an mg/m‘ basis with what
`would have been found on an mg/kg‘ basis.
`Some indication of this has already been given
`in chart '7 which shows that there is a close
`relationship between 12 times the LD10 in the
`BDF. mouse and the MTD in man. Since the
`relationship between mg/kg and mg/m“ used is
`(Inc/m‘) = (km)«>< (ms:/kg).
`(i = 1,---.7),
`models (1) and (2) become, in terms of mg/kg,
`_
`(/cm)”
`(dose inman) = mm)“
`X (dosein animal system)
`and
`
`(1)
`
`(lcm)i.
`_
`a
`(dose in man) = mm)“
`(2)
`x (dose in animal system)
`where (7cm)u and (lamp. refer to the (ma) fac-
`tor in the particular animal system and man
`respectively, and A1 is exactly the same as
`stated before. Hence it should be clear that dose
`in mar. can be predicted equally well either on
`'1 mg‘/kg basis or on an mg/m“ basis. Thus by
`ing the ma factors and model (1), the dose
`man (mg/kg) is approximately 1/,2 the dose
`i mice,
`1A, the dose in hamsters, 1/7 the dose in
`rats, 1/3 the dose in rhesus monkeys, and 1/2 the
`dose in dogs.
`
`DISCUSSION
`
`implied for
`Originality is not claimed or
`this analysis. We have confirmed and extended
`the general observations and conclusions of
`
`VOL. 50, N0. 4, MAY 1966
`was cepieei,
`cl :m..a§r he
`miarigglit La ws
`
`Pinkel (2) Who confirmed and extended specific
`8SI>ects of the basic observation of Rubner
`(36), made 80 years ago, and many other inVeS_
`tigators later.
`
`The availability of much more extensive
`toxicity data from the Cancer Chemotherapy
`National Service Center program, from certain
`other published sources, and from our own labo-
`ratories seemed to make this present analysis
`timely. Also we believe it is important to use
`more definitive biologic end points of toxicity_
`Th_15 ilfléllysis and study of data on toxicity to
`animals and humans of several types of anti-
`cancer agents (tables 1, 3, and 5)
`lead us to
`conclude that the toxic dose of an agent
`is
`similar among species when the dose is meas-
`ured on the basis of surface area. The skin sur-
`face area was used here though it is unlikely
`that the skin is he target area of action of any
`particular drug. More likely the skin surface is
`more or less proportional
`to the true target
`surface.
`
`that mammalian species are
`To the exteii
`broadly similar and have corresponding organs
`and tissues, it is true that any surface area will
`increase approximately with the tWo.thi1‘dg
`power of weigit
`(38). Thus the two—thii'ds
`power of body weight would have been a con-
`venient unit of surface area to use and the re-
`sults of the analysis would have been almost
`the same (see A pendix II).
`Pinkel
`(2) suggested that “cancer chemo-
`tlierapists consider the applicability of body
`surface area as a criterion of drug dosages in
`their laboratory and clinical studies.” We sug-
`gest that a uni proportional to body surface
`area is sufficient and an appropriate unit is
`(weight) “".
`We have been conccriied only with compari-
`sons among species, not within species, and
`with adult animals, not
`immature and adult
`animals. Also we have been concerned solely
`with anticancer drugs.
`Some of the toxicologic data tabulated may
`disagree with unpublished and published obser-
`Viltlofls Of Some Oxllerimeiitalists and clinicians.
`The Acute Leukemia Task Force of the Na-
`tional Cancer Institute wishes to correct, up.
`date, and extend this analysis at some future
`time. Those interested in seeing such correla-
`tion eiforts extended can help by providing ad-
`225
`
`|nnoPharma Exhibit 1062.0009
`
`

`

`ditional data, both clinical and experimental, in
`a form similar to that in table 1.
`The present study has emphasized the quan-
`titative aspects of toxicity of anticancer drugs
`to animals and man. Regarding the prediction
`of the qualitative eifects of anticancer drugs
`in man from laboratory animal studies, Owens
`(1) suggested:
`Predictive value
`Good
`Questionable
`None
`
`P-rccliaical toxicity .s‘f'l(.ll’l:ZS
`Bone marrow, gastrointestinal tract,
`liver, kidney
`Nervous system,
`including periph-
`eral neuropathy, extraoeular pal-
`sies, and CNS toxicity
`Skin and appendages, including skin
`rashes, dermatitis, and alopecia
`
`Of the 18 agents in this study, 17 produced
`limiting toxicity to the bone marrow (marrow
`depression: MD) and to the gastrointestinal
`(GI) tract. If the mg/ni’ doses in man that are
`predicted by using the weighted combined esti-
`mate are compared to the observed doses, then
`the largest ratio of predicted dose/observed
`dose is 3, for thioTEPA. Consequently it would
`be reasonable to study preclinical toxic effects
`in the mouse, rat, dog, monkey, and hamster,
`to estimate the MTD (mg/in‘) in man, and to
`start clinical cancer chemotherapy trials at
`about one-third
`the
`predicted
`dose. This
`would have been a safe procedure for all 18
`drugs mentioned. Owens (1) suggested that it
`might be reasonable “to begin a human trial at
`one-tenth of the maximum tolerated dose in the
`most susceptible animal" (on an mg/kg basis).
`Since the most susceptible animal will ordi-
`narily be the dog or rhesus monkey, Owens’
`rule of thumb on an mg/m“ basis becomes:
`begin trial in man at about one-third the close
`for monkeys or one-fifth the dose for dogs.
`Thus there is reasonable agreement between
`the two recommendations. However if the ani-
`
`inal data are not placed on the mg/mi
`before using Owens‘ rule of thumb, any ."m_1IS
`tional knowledge which the small
`ztlilovérl
`(mouse and rat) might contribute will be [dues
`looked. Remember also that the toxicity V;n0re
`reliable statistically because more anima -
`<
`(LD10’s) for such small animals are Often“ we
`
`_/m,
`_
`_
`generally used.
`The ratios of animal/human toxicity (mfxmm
`basis) for the mouse, hamster, flog, and each
`key are remarkably close to unity.’ Thllbhig 1,‘:
`species generally predicts for man. Phat tnfi ta
`true for the mouse is D€L1‘tiCUlé11‘1y Pe1'm“’Hl
`_
`cancer chemotherapy. Extensive drug‘ dfi‘ feolfin
`merit programs ‘which use mouse tumois hue] ,
`to be on firmer ground than we had p1‘eV10/msl-:3
`thought. In general the rat is more suscefl Ti e
`to these agents than the other 51199105-t
`_,1
`hamster is unusually resistant to amet_h°_1(’l.eU?1
`and sensitive to the fluorinated DY1'1m1I“V1eb'
`The dog and monkey, long kn(.>vi_/Y1 t0 be 1-"5"‘%°""
`ably good predictors of
`toxicity '99 humam’
`have shown up well
`in this anfllY51_S-
`t I
`We are not suggestiiig that it is wise to age
`mouse or rat LD10’s, convert
`the doses
`0
`mg/m’, and then start clinical trials zitrgélef
`third this level (in mg/m‘ for i1‘1f1T1)- The
`1'
`tional safety provided