`
`MAY 1966
`vol. 50, no. 4
`
`U-S- DEPARTMENT OF HEALTH, EDUCATION, AND WELFARE
`
`o
`
`Pub 10
`
`l' H ltl Service
`
`ea
`
`1
`
`I
`
`InnoPharma Exhibit 1062.0001
`
`

`

`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.
`
`Submit an original and 2 carbon copies of the manuscript, typed double spaced. Each page should begin with
`a new paragraph.
`Use accepted spelling, abbreviation, and hyphenation found in Webster’s New International Dictionary.
`Footnotes should be typed at the bottom of the page on which they appear.
`Tables
`
`Submit each table on a separate page, typed double spaced. Please use standard abbreviations; otherwise spell out.
`References
`Submit references typed double spaced. Follow the standard form of Index Medicus: author, title, name of 50111“
`nal, volume number, page 01‘
`inclusive pages, and year.
`In book references, give author,
`title of book
`edition, if more than one, city, name of publisher, date of book, volume number, and pages cited. References
`in text should begin with number 1 and continue consecutively.
`.
`Illustrations
`Photographs and graphs preferably should be clear, glossy prints. Original art work should be done in black ink
`for good reproduction.
`Nomenclature
`
`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—
`viations of journal titles in references.
`.
`Only standard abbreviations are acceptable. If a new or coined abbreviation is used, it must be defined when first
`mentioned.
`
`.
`
`,
`
`Reprints
`
`Supplied free of charge: 250 reprints will be sent to the senior author about 1 week after publication of the article.
`
`Th is material was partied
`at the NLM and may be
`Su Dian Uzifieeyright Laws
`
`InnoPharma Exhibit 1062.0002
`
`

`

`r“QUANTITATIVE COMPARISON OF TOXICITY OF ANTICANCER AGENTS
`IN MOUSE, RAT, HAMSTER, DOG, MONKEY, AND MAN 1;;
`Emil Jil:reireich,3 Edmund A. iGehan,‘ David P.1RG”,5 Leon H.'Schmidf,6 and Howard E.'Skipper7
`
`SUMMARY
`
`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/m“ of surface area). Two models were assumed for the relationship
`between the maximum tolerated dose (MTD) in man and the approximate I
`LDlO in each animal system:
`(dose in man) = (dose in animal system i)
`
`(1)
`
`and
`(2)
`, 6)
`,
`,
`,
`(dose in man) = A.- X (dose in animal system 2'), (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, BDFl 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/m2 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 (km). is the appropriate factor for converting doses from mg/kg to
`mg/m2 surface area for each species. When the (lamb factors are known,
`equally good predictions of MTD in man can be made by either dose unit.
`On an mg/n'l2 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 1/12 the LD10
`in mice,
`1/9 the LDlO in hamsters,
`1/7 the LDlO in rats,
`1A} 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 tox1cities of potential
`anticancer drugs correlate remarkably closely With the results in man.
`
`,
`
`I
`f
`
`i
`L
`(
`i
`
`I
`'
`
`I
`
`
`
`,-—..——-v——~.r4
`
`-
`
`vised Jan 17, 1966.
`
`5Laboratory of
`
`.Chemical Pharmacolo
`
`, National
`
`1
`
`.
`
`‘
`
`J
`[I
`
`zlstfsgyeélofeecuiggiffé Zispices of the Acute Leu-
`Sigma 'IlrdSt’ Public Health ‘Service, Bgtihesda, Md.
`kemia Task Force of the National Cancer Institute by
`ase f“ rcss iequests for reprints to Dr. Rail.
`the Subhuman Subcommittee.
`0National Center
`for Primate Biology, Univ of
`.
`3 M. D. Anderson Hospital, Houston, Tex.
`California. at Dav1s.
`‘Biometry Branch, National Cancer
`Inst, Public
`Kettering-Meyer Laboratory of Southern Research
`i Health Service, Bethesda, Md.
`Inst, B1rmmgham, Ala.
`I
`CANCER CHEMOTHERAPY REPORTS VOL. 50, N0. 4, MAY 1966
`ll
`
`219
`
`“i This material may be protected by Copyright law (Title 17 US. Code)
`_ A
`l
`
`I
`
`InnOPharma Exhibit 10620003!
`
`

`

`eutic
`No attempt was made to relate theta?“ the
`doses in the various mammalian specletsgemptcd
`future this correlation should be hast is not
`since the therapeutic target in the
`0.01, if an
`the same as the toxicity tarse?- Howe; experi-
`agent has therapeutic DIODQTUCS 1n alose level
`mental system, it is well to know the (tification
`for patients. Since there is some nutshese (1059
`for using MTD’s in cancer therapy,
`levels were studied.
`.
`.
`as to
`The plan of this retrospectl‘f‘3 hfgdfigzained
`examine considerable toxicologic‘fldaiy screen-
`in (a) small animals, used in Dllénflg evalua-
`ing and quantitative secondary
`Ionkcys, for
`tion; (1;) larger animals, dogs, and m as 0f tox-
`the quantitative and qualitative {iszpfd “(0) man,
`icity at sublethal and lethal levels, :6” determine
`the target species. The 22031 was Otween cer—
`What relationship exists: if any, b:
`oints in
`tain commonly used toxicologic en f I; a num-
`the Various animal species and man 0
`her of anticancer agents.
`.
`(r est
`Nothing in this report is intendedlh: 11:539.
`or imply that short cuts are allow/EH 3 Dose-
`clinical or clinical
`toxicologlc {Stu El‘m,enot
`limiting and serious toxic effects in ”ft carefully
`always apparent from even thc.mf)5'mals (1),
`done toxicologic investigations in 11111.17 under—
`It is emphasized and should be clam Jt
`to em-
`stood that it is dangerous to attemipity dam
`trapolate directly from animal
`’orxNCew drugs
`to maximum tolerated doses m mild t ial only
`can be introduced safely into cllmcffl
`racologic
`through careful toxicologic and phatrmuq study
`study in animals and then very can 1? éosages
`in man, starting with much 10W‘31 (1 b y the
`than those which appear to be tolerate
`3
`animals.
`DY
`APPROACHES AND ASSUMPTIONS IN THIS STU
`‘ h
`-
`The published and unpublished datgbggfigd
`form the basis for this analysis W‘mierent mm
`bv numerous investigators 115mg dii gtent and
`tocols and end points. We used (101151: the data
`reasonable general assumptions so tha.
`ts pro—
`were comparable. The biologw end iponées’ssary»
`to make the results more compam e
`tocols, assumptions, and correctionsbllle are do:
`scribed briefly.
`.
`Toxicologic End Points (See Appendix I)
`- h
`,
`.
`.
`Mouse
`rat, or hamster: Lethality—lhzng03;h‘;illfic
`when administered by a certain rout'eh LD10) dur-
`killed a selecteid Ii’encentéige gist-misc :0 more than
`'
`\
`ifier
`o servn ion
`“ ,
`.
`_ n“
`‘1th 3111513155 were used in a typical dctermmaho
`CANCER CHEMOTHERAPY REPORTS
`This material in
`at the NLM and
`Euhjefit US; 43:}:
`
` l
`
`The biologic aspect of a drug development
`program to
`discover
`compounds
`effective ,-
`against any clinical disease is
`generally an ex-
`macology. In the
`ercise in comparative phara
`typical program, compounds are screened in
`small animals against some easily produced and
`reproduced pathologic condition A close rela-
`tionship must exist between t
`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 in 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). Hopefully two benefits might‘accrue
`from this evaluation: (1) If there is reasonable
`consistency in the reactions of various mam-
`malian species,
`the toxicologie 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
`approached 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
`
`InnoPharma Exhibit 10620004
`
`

`

`typically 274 animals
`(c) MTD;
`Dog or monkey:
`were used at each dose level, spaced by 2—fold incre—
`ments.
`In all
`instances individual doses which killed
`0 and 100%; were used. The highest dose killing 0%
`was considered the MTD.
`(b) Dose-related, hema-
`topoietie ell‘ccts;
`localized hemorrhages of the gastro-
`intestinal tract; generalized hemorrhagic lesions (ab-
`dominal and thoracic viscera); stimulation of the cen-
`tral nervous system (CNS) ; others.
`Man:
`(a) M’I‘D for a fixed schedule (dose causing
`mild to moderate sublcthal toxic effects in a significant
`percent of patients);
`(5) MTD for a variable sched<
`ulc, calculated from the daily dose and median period
`to toxic eilects requiring 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-
`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 very much from the dosl
`age that should have been used.
`It Was necessary to use toxicologic data ob-
`tained by various routes of drug administra-
`tion, ic, intraperitoneal (in) 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 iv
`routes are usually compa ‘able.
`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.
`Griswold et a1.
`(3)
`reported that when the
`LDIO’s in BDF. mice of 70 agents, including
`the major classes of anticancer agents, were
`compared for two schedules, qd 1—7 days and
`qd 1711 days,” the mean ratio'(qd 1—7 days/
`qd 1—11 days) was 1.56. This is very close to
`that which might be expected from direct cumu—
`lative drug toxicity (ll days/7 days = 1.57).
`Pinkel
`(2) and other investigators pointed
`out that the usual doses of certain drugs in
`various animal species and lnan were compara-
`ble when the dose was measured on the basis
`of mgr/mz of surface area. Consequently most
`of the results are presented in mg/m’. However
`since rug/kg is a commonly used unit of drug
`
`dosage, some results are also presented in this
`“qd = drug given once daily for as many days as
`indicated.
`
`
`
`l
`
`VOL. 50, No. 4, MAY 1966
`was cool ed
`1d may he
`myrigIh-t Laws
`
`unit. Only a simple transform
`ation is required
`herefore the rela-
`to change rug/kg to mg/m‘; t
`tionships developed are equivalent whichever
`unit
`is used. The quantitative relationships
`were Simpler when expressed in mgr/m2
`A conversion factor (km) was used to trans.
`form mg/kg to mgr/m2 by the equation mg/
`kg ><
`(km) : mg/m’; (km) factors for ani-
`mals, given their weight, are presented in table
`1
`(Appendix II), and table 2 (Appendix II)
`presents a way of transforming doses in mg/
`ikg to mg/m’ for man, given height and body
`weight. Chart 1
`(Appendix II) is a diagram
`for determining 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, possi»
`bly related to length or some organ-membrane
`surface area, would be as appropriate or more
`appropriate. However the body surface area has
`been used to relate many physiologic param-
`eters among species and means of transform-
`ing the data are readily available. Further, in
`our clinical studies we routinely use body sur-
`face area to adjust drug: dose for patients of
`different size and weight.
`RESULTS
`The first step in analyzing 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 1710 days, we calculated that
`the LD10 (or MTD) for a schedule of (1d 1—5
`days was twice that value. The next step was to
`
`convert doses (LDIO‘s or M'I‘D’s) from trig/kg
`to mgr/n1" This was accomplished by the ap«
`proximate formula
`, 7)
`.
`.
`(mg/kg), (5:1, .
`><
`(mg/m“) = (Ion).
`where the (km). factor difl'ers according to the
`species and also according to body weight with—
`in each species.
`In the analysis an average
`(km)i factor was used, assuming that individ<
`uals in each species were of average height—to—
`body-weigrht
`ratios, The (km).
`factors were
`derived from standard relationships between
`weight and surface area as given in Specter
`(40) and Scndroy and Cecchini
`(39). Details
`and other information on relating drug doses
`in mgr/kg to doses
`in mg/in2 are given in
`Appendix II.
`
`221
`
`InnoPharma Exhibit 1062.0005
`
`

`

`o
`
`CHART a
`Comparison al ioxmly dam on onncancw
`agents in the mouse and manlon a
`MG/M2 bums}
`
`O Annmambomes
`A Alkylmmg agents
`
`I Olners
`
`O
`
`
`
`
`
`*4————i—7#—‘
` J
`
`I!)
`lOO
`IOOO
`01
`LG
`eon nousmowlmc/Mzzool-soayscncdne)
`
`’3 1000
`:3
`“é
`:2
`;
`f [00
`CD!
`N;
`<3ELu
`8
`ac:w>—<mLu
`6‘
`:
`gEX<1
`2.2
`§
`
`IO
`
`o.|
`
`10
`
`
`
`“fir
`
`l000 r-
`
`IOO ,
`
`CHART 3
`Comparison 0! toxicity dam on anticancer
`uqenls is! We humus! and mon(on a
`MG/M2 basis]
`0 Anhmelahahles
`A Alkylahnq uqenis
`0 Omevs
`
`
`O
`
`O 0
`
`.
`
`
`
` J
` | 0.0
`
`|.0
`
`oil
`
`0|
`
`“J
`IOOO
`
`I00
`I00
`10
`HAMSTER‘LD‘O(MG/iflztoul-Sbayschedule]
`anima systems. We wish to describe the rela-
`tionship between the dose-toxicity curve for
`man and that for each of the animal systems,
`Two models are considered:
`(dose in man) : (dose in animal system i)
`(i=1,...,6)
`(l)
`and
`(dose in man) : A X (dose in animal sys‘
`temi), (i:1,...,6).
`(2)
`Model
`(1)
`is a special case of model
`(2)
`since they are the same when A. : 1. Model
`CANCER CHEMOTHERAPY REPORTS
`
`"Chemical Abstracts’ nomenclature and NSC num-
`bers for the agents are given on page 243.
`222
`
`Th material we
`at. the HEM and
`
`Emmett U33 Ctr-pg
`
`InnOPharma Exhibit 1062.0006
`
`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/m2 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
`
`
`
`MAMMAXlMUMTOLERATEDDCSE[MG/Mz‘ODl-5Doyschedule)
`
`
`
`
`
`[000
`
`lOO
`
`CHART l
`O
`Cflmpmlson of 101m” dam on unlicnncer
`agents (or the Swiss mousennd man
`
`
`[on u MG/Me basis]
`0 Ilnnmetahcllles
`
`A Alkylmingngems
`o Olhers
`
`
`
`
`
`5
`
`‘o
`
`0
`
`0..
`
`IOO
`l0
`LO
`SWISS MOUSE'LDI0 (MG/MZ‘ 0D l-S Day schedule]
`
`I000
`
`
`
`
`
`
`
`
`
`MAMAXMUMTOLERATEDDOSElMG/MZIOUI-5Dayschedule)
`
`

`

`CHART e
`
`'000
`
`'00
`
`Campunscn oi loxmly data an anticancer uqems (m
`me dcqcnd moMon a MG/MZ bums]
`C) Anhmelabomes
`A Alkylalmq aqenls
`. Cihels
`
`0
`
`A
`
`6
`
`5
`
`___l
`IDOO
`100
`ID
`10
`0.1
`DOG: MAXiMUM TOLERATED DOSE lMG/MZJOD #5 Day schedule)
`CHART 7
`Campanscn uv manuals on unhcuncey agents to!
`(he mouse and man (nnn MG/KG Deals)
`The lZil velnhcnsmp shown en 0 MG/KG busts KS equwuienl
`lo we H ralunansmp shown an MG/MZ buss (cm 2)
`The nvproxxmule 121$ VEIG'IGHS'HD(MOUSEIMUfillS m
`agreement wh the who cl lhe KM factors used
`‘0 _ fm Mesa specle5.ic,37i3lmun.muusel:cu|2
`0 Antmclubolnes
`A Alxylanng ugems
`. omevs
`
`o
`
`0
`
`O
`CA
`
`m
`QQ
`
`A
`
`10 7
`
`O,| -
`
`’6 WC ,
`E
`i:
`3
`2
`._
`
`a g
`
`BDF‘ MOUSE, LD‘0 [MG/KGVQD l-5Day schedule)
`
`raised, assumed to be 1 in models (I) and (2).
`This model is a natural generalization of (2).
`However, since the estimates of B; were near
`1
`for all animal systems,
`in fact within 1
`standard error (SE) limit, there is no advantage
`to using 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 sump-
`ling variation (‘and other assumptions as men-
`223
`
`InnoPharma Exhibit 10620007 ‘
`
`
`
`
`MANIMAXIMUMTGLERATEDDUSEIMG/MZ-ODPfiDayschedule] 0!
` OOl -———J
`0m
`
`f
`g
`
`W8
`
`aamEmm_
`
`J:>
`”
`1Ez4s
`
`2gi<
`
`CHART 4
`Comparison OiiDXlCiiy dam an anticancer
`agenls for the m0 and munlnn 0 MG/ A
`
`News's)
`
`O Annmelubames
`
`/\ Alkylchng ugems
`%
`C Others
`
`
`
`
`O
`
`
`
`1000
`
`loo
`
`
`
`(1{‘5Guyscheduiel
`
`‘9,
`
`moc:
`
`2Lu|..(1acw.1or/
`
`
`
`
`O.
`
`1
`
`l_______lri—l
`
`OJ
`1 O
`Ifl
`IOD
`lOOO
`RAT: LDm (Mo/M2410 1-5 Day schedule]
`
`CHART 5
`(9
`Compansan of Iamity dam on unllcantcr
`
`
`agents {or ihe rhesus mnnkey and man
`0
`(on u Mc/M2 basisl
`A
`
`0 Anhmeiubcllies
`
`A Alkylullng ugenls
`' Ovhers
`
`
`
`
`g IOUO
`fw’
`E
`
`mL
`
`5
`
`loo
`
`
`
`I 0
`
`_l.,.__...-'.li_—__,..J
`I0
`l00
`lOOO
`RHESUS MONKEY:
`MAXIMUM TOLERATED DOSE(MG/MZ.QD I-EDnyschedule)
`
`
`
`(1) assumes that the dose in each animal sys-
`tem gives a direct predictlon 0f the (lose in
`man. Model (2) assumes that the dose in man
`is a fraction (.11.) of the dose in the animal
`gystem and the fraction remains constant for
`the sample of drugs.
`A third model was considered:
`ose in man) 2 A. X (dose in animal sys—
`tem i) ”4,
`(i = 1,. .. ,6)
`is the power to which the dose is
`
`(d
`where B.
`
`voL. 50, N0. 4, MAY 1966
`I was {spied
`rid may be
`rpy‘r‘igh-t La WE
`
`
`
`

`

`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
`Al : 2 SE are given in the bottom half of table
`4‘ Note that the approximate 95% confidence
`limits for the multiplying factor, 11., 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 (I) 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
`
`”’00
`loo
`
`10
`
`OJ
`
`
`
`
`
`MANrPREDlCTEDDOSAGE(M6/M2)
`
`CHART 8
`
`-
`Z
`Observed momma dosegesiMG/M human
`usmq all unimuisysiemsiweiqhied estimates]
`0 Model
`1
`0 Model 2
`0
`
`.
`/9
`.
`o
`(g.
`O
`
`35/;
`
`
`
`I002
`10
`|o
`MANlOBSEHVED DOSAGE [MG/M )
`
`O.I
`
`in table 5 and the weighted estimates base: 0;:
`all animal systems combined are plo ‘3“ as
`chart 8. The best estimates of dose 1T1 mam; 4
`indicated by the standard devlz'ltions m :ématesl
`are given by weighting the indivxdual ES 1
`from each animal system.
`.
`_
`.
`Another model was considered in which tlil:
`dose in man (mg/m”) was related to doses
`the animal species in a single equation.
`
`‘0‘; (dose in man) : 0.284 + 0.847 10g (dose In
`Swiss mouse)
`.
`—— 1.064 log.r (dose in BDF‘,
`mouse)
`'
`+ 0.539 log (dose in rat)
`»I— 0.801 log (dose in men—
`he )
`.
`— 0.13775 log (dose in dog).
`
`This predicting equation leads to a slight im:
`provement in the prediction of the dose in man ,
`the deviations of observed from Pl'edlCtei610$-
`ages were less (standard deviation of 0._2 lit on
`log scale compared to 0.275 by usmg weig 6d,
`combined estimates). However a VFEdICtiOn 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 L6, this qiiestlor
`arose: Do the differences between the Close:
`
`CANCER CHEMOTHERAPY REPORTS
`This material i
`at til 5 NLM am
`EUithEELt US {in}:
`
`InnOPharmé 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 overpredict
`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 EDF, mice the predictions of
`dosage in man were lower for antimetabolites
`than for alkylating agents. There was no evi—
`dence of a difference in the other species. Only
`4 antimetabolites 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 rag/m1 basis with what
`would have been found on an mgr/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
`(mg/m“) = (lamb >< (mg/kg),
`(73 = 1 ,
`.
`.
`. ,7),
`models (1) and (2) become, in terms of mg/lig,
`
`Horn)“
`.
`(dose in man) = (107%)“
`X (dosein animal system)
`and
`
`(km).
`_
`(dose in man) = “MUM A.
`(2)
`x (dose in animal system)
`where (km). and Ham)". refer to the (km) 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 man can be predicted equally well either on
`“- mgr/kg basis or on an mg/m” basis. Thus by
`ing the km factors and model (1), the dose
`man (mg/kg) is approximately 1A2 the dose
`1 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.
`
`(1)
`
`DlSCUSSlON
`
`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
`4
`was copied
`of may be
`
`upright La we
`
`Pinkel (2) who confirmed and extended specific
`aspects 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.
`This analysis and study of data on toxicity to
`animals and humans of several types of anti-
`cancer agents (tables 1, 3, and 5)
`lead us t0
`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.
`
`
`
`To the exten that mammalian species are
`broadly similar and have corresponding organs
`and tissues, it is true that any surface area will
`increase approximately with the two~thirds
`power of
`\veigit
`(88). Thus the two-thirds
`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-
`therapists 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 concerned 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-
`vations of some experimentalists 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 efforts extended can help by providing ad-
`
`225
`
`InnoPharma Exhibit 1062.0009
`
`

`

`ditional data, both clinical and experimental, iii
`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 eifeets of anticancer drugs
`in man from laboratory animal studies, Owens
`(1) suggested:
`PredicHoe value
`Preclinical toxicity studies
`Good
`Bone marrow, gastrointestinal tract,
`liver, kidney
`Nervous system,
`including periph-
`eral neuropathy, extraocular pal-
`sies, and CNS toxicity
`Skin and appendages, including skin
`rashes, dermatitis, and alopecia
`
`Questionable
`None
`
`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 nag/m2 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/m‘) 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/m2 basis becomes:
`begin trial in man at about one-third the dose
`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/m“ 1:331:
`before using Owens‘ rule of thumb, any "111115
`tional knowledge which the small anl Vgrl
`(mouse and rat) might contribute will be E>1
`s
`lookEd. Remember also that the tox1c1ty Vdiuee
`(LDlO’s) for such small animals are often n Oi‘e
`reliable statistically because more animal:- 51
`generally used.
`.
`.
`2
`The ratios of animal/human toxic1ty (mg/1:11—
`basis) for the mouse, hamster, dog) and {30,11
`key are remarkably close to unity; ThusI $101.4
`species generally predicts for man. [hat ’6 1513‘»)
`true for the mouse is particularly pertinef:l
`cancer chemotherapy. Extensive drug devt‘ 013'
`ment programs ‘VVlllCll use mouse tumors >981“:
`to be on firmer ground than we had preVIOtLlf 1.;
`thought. In general the rat is more suscep ’ll‘l e
`to these agents than the other speCies.
`.-1
`hamster is unusually resistant to amethoil‘fiellfl
`and sensitive to the fluorina’ced pyrimidllleb-
`The dog and monkey, long known to be redbon"
`ably good predictors of
`toxicity to humans,
`have shown up well
`in this analysis.
`We are not suggesting that it is Wise to take
`mouse or rat LD10’s, convert
`the doses to
`mg/m’, and then start clinical trials at One—
`third this level (in mg/m‘ for man). The addi—
`tional safety provided by toxiCity datadiorn
`multiple species is well established, as 25 the
`value of specific qualitative knowledge on dose—
`related sublcthal toxicity and its reversibzltty.
`Finally it is suggested that the quantitative
`relationships between toxicity to animals and
`to humans are simpler w