Abstract
Tobacco smoke contains nitrosamines, which, according to this 1963 letter to Helmut Wakeham (vice president in charge of research at Philip Morris) are "the most potent carcinogens known." A report published prior to this time, in 1962, in the journal Toxicology and Microbiology (a copy of which was also found on the Philip Morris site) entitled "N-Nitroso Compounds and their Precursors in the Environment" states that humans get "life-style" exposure to nitrosamines from tobacco and tobacco smoke. A quote from that report says:
"The concentration of nitrosamines found in tobacco products represents the highest human exposure to N-nitroso compounds....To date, over 20 different preformed N-Nitroso compounds have been found in tobacco products..." (Philip Morris site, Bates No. 2060547756/7784, URL http://www.pmdocs.com/getallimg.asp'if=avpidx&DOCID=2060547756/7784
Today's document is a letter from a researcher in Switzerland who attended a meeting to determine the "state of analytical methods to disprove [the presence of nitrosamines] in tobacco leaves and smoke." The scientist writes to Dr. Wakeham stating:
"I thought the importance of the [study on nitrosamines] is such that you might be interested in an immediate short notice of the highlights of the problem discussed...
Some 60 Nitrosamines -- as well as Dialkyl as more complex and assymetrical compounds -- have been tested. They are all carcinogenic and what is more organo-specific. According tot he compound used and according to the way of application (intravenous, oral, sucutaneous), cancer is produced in a given site such as lung, liver, bladder, oesphagus, stomac and also for the first time in history, in brain...The dosage needed to induce cancer in test animals is for most Nitrosamines exceedingly low. In some instances a unique application is sufficient....Hamsters on inhalation with tobacco smoke after 8 months show lung lesions (there is reasonable belief that in several more months lung carcinomas will be obtained). Under similar (not identical) conditions lung cancer have been obtained with Nitrosamines...As a whole one can say that the Nitrosamines are very potent carcinogens, potent mutagenes, that they have a very good dose-response relationship, an astonishing relation between structure and organotropic action, that their effect on the chemical structure of the attacked organism is better known that for most other carcinogens..."
Fields
- Notes
This document was used as a trial exhibit in Minnesota's case against the tobacco industry, as well as more recently in the Boeken case in California.
- Quotes
Dear Mr. Wakeham,
Lat week, we had a working-conference in Hamburg concerning Nitrosamines and Lactones, their carcinogenic action upon the animal and the state of analytical methods to disprove their presence in tobacco leaves and smoke.
The meeeting was attended by Prof. Druckrey of Freiberg--who took the initiative to organize this international get-togther between industry and cancer research-: Prof. Dickens, Courtwalk Institute of Biochemistry; Prof. Dontenwill from Pathology Department of Munich; Prof. von Euler stockholm; Dr. Kriek, Antoni van Leewehoek-Huis (Dutch Cancer Institute); Dr. Magee and Dr. Heath, Medical Research Council, Carshlaton, England; Dr. Roe, Chester Beatty Res. Institute; representatives of the German hygienic office, various collaborators of the mentioned persons and representatives of Swedish, English, German, Austrian, Frech and Swiss tobacco manufacturers.
Although proceedings will be published in due course, I thought that the importance of the matter is such that you might be interested in an immediate short notice on the highlights of the problems discussed. ..
Nitrosamines
Magee, Roe, Druckry, Emelot and Kriek have all worked extensively on the subject.
Nitrosamines are the most potent carcinogens known. By oral, subcutaneous or intravenous application tumours are obtained at various spots, usually far from the site of application.
1. Active Nitrosamines and site of action
Some 60 Nitrosamines -- as well as Dialkyl as more complex and assymetrical compounds -- have been tested. They are all carcinogenic and what is more organo-specific. According tot he compound used and according to the way of application (intravenous, oral, sucutaneous), cancer is produced in a given site such as lung, liver, bladder, oesphagus, stomac and also for the first time in history, in brain.
Without going into details of what kind of cancer is produced by which Nitrosamine, it is interesting to note that of the symmetrical Nitrosamines only the Diamyl-compound produces lung tumours....
2. Dosage
The dosage needed to induce cancer in test animals is for most Nitrosamines exceedingly low. In some instances a unique application is sufficient. In one case, a unique intravenous application of 100 mg Methyl-nitroso-urea produced kidney tumors.
3. Animal metabolism
The Nitrosamine distributes quickly through the test animal and is completely metabolised after 24 hours...
4. Tested animals
Rats, mice monkey, fishes, golden hamsters gave all similar positive responses.
5. Proposed pathway of action
It is thought that the alkylating agents, through alkylation of DNA, can alter the nucleus and in such a way be responsible for the start of cancerous growth....
...8. Inhalation
Hamsters on inhalation with tobacco smoke after 8 months show lung lesions (there is reasonable belief that in several more months lung carcinomas will be obtained). Under similar (not identical) conditions lung cancer have been obtained with Nitrosamines.
9. Nitrosamines in relation to tobacco
a) Presence
The presence of Nitrosamines in tobacco or smoke is not established, nor disproved, for lack of a suitable analytical method. One method is in elaboration in collaboration between Prof. Druckrey and labs of the German industry....
10. Conclusion
As a whole one can say that the Nitrosamines are very potent carcinogens, potent mutagenes, that they have a very good dose-response relationship, an astonishing relation between structure and organotropic action, that their effect on the chemical structure of the attacked organism is better known that for most other carcinogens....
- Company
- Philip Morris Cos., Inc.
- Author
- Waltz, P.
- Recipient
- Wakeham, H.
RegionSwitzerland
United States
LitigationStmn/Produced
Stmn/Selected
Stmn/Trial Exhibit P-11607
Stmn/Trial Exhibit P-2502
TypeBibliography
Letter
Scrt, Report, Scientific
SubjectHealth
Health Effects
Toxicology
Animal Subjects
cancer
Document Images
Page 1: mry24e00
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M
r. H . _.LJ
akeham . o _ .
Vice Presidentfin charge of Research
Philip'Morris International
P.O...Box 1.895
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September25, 1963
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Personal and confidential
ear Mr. Wakeham.
Last week, we had a working-conference In Hamburg
concer.n:ing__Nitrosamines and_Lactones, their-carei-
_ . ._,
nogenic action upon the animal and_the state_.of_
analytical methods to disprove or prove_their,_
nresence in tobacco leaves and smoke.
e.meeting was attended by Prof.
Th
D
~ ruekrey of
,
TFreiburg - who_took the initiative to organize this
international,.get,together between indu$try and can-
cer,"research -; Prof. Dickens_, Courtauld Indtitute
of Biochemistry; Prof. Dontenwirll_ _f,rom, Pathology
Department _ of Munich;- Prof. von Euler Stockholm;
Dr.,,Kriek,__Antoni van.Leeuwenhoek-.Huis (Dutch Cancer
Institute); Dr. Magee and Dr. Heath, Medical Re-
reh
sea ,Council, Carshalton, England; Dr: Roe, Chester
Beatty Res. Institute; representatives_of the German
hygieni,c office, various ,collabo,rator,s Qf _ theqmen-,... ,.
,tioiied+pens and representatives ofScledish, English,
_
German, Austrian, French and Swiss tobacco manufac
turers.
-
:;A1_tho,ugh proceedings will be published in due course;
'',I .thought +t~~t the importance_ of the matter is such
that you might be interested in an__immediate short
notice on the highlights of the problems discussed..
a .: ..:. , . ,.~
.,.-
~
co
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Lactones
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Prof, :=.Di-cken.s showed _results contained partly in his
paper'(l). Contrary to_his expressed_opinion that
narcinogenic ,Lactones must r?actq,,with Cys,teire in
c
eut_ral_-soluti.ori, he found substances -lik.e..,Aflotoxin B,
--
reacting only pooly with Cysteine, but.being highly
carcinogenic.
However .,t,here ae®ms,_to,_#be- good proof that the car-
_
,. ..
_cinogenic action :af_Lact,ones is,associated with the
alcylation-of the.SH-,group___of_Cysteine in_the Nucleo-
:..:.proteins_of the ce11. Active factors are not yet shown
>=in tobacco_or.smoke (Cumarirn is not carcinogenic).
Sincethewact,i,wonof Lactones. is -on the site of appli-
cation(,local-,tumours._only), Dickens-proposes to seek
Lactones rather,_,in ,toba.cco, associating a; possible
presencem of Ldctones with _cancer of nthe mouth_ in
~=Indian tobacc,ochewers. Lactones_have no carcinogenic
action far fromthe_ spob of injection.
.
WThe Oxy _alfa-zalerolactones_,- presence of which was
yshowx~ r,in t,obacc,o (3ntralblatt, 789, (1938 )). has not
been tested fo.r,c,arci,nogenic action, nor the Lactones
found _by Reynolds (Tetrahedron, 246, (1961) and 107,
(1g63)) which showed the typical double-bond next to
the ~ Lactones group, apparently necessary for carcinogenic
~...
a'c'tion.
fi
Magee, Roe, Druckrey, Emel.ot_and_Kriek have all worked
;extensively_ on the- subject.
Nitro.samine_s are the most_potent car
cinogcns known.
,By oral, subcutaneous-,or._intrayenous,application
tumours are. obtained at, v_ario_us, _spotit, usually far
`from the site of application.
Page 3: mry24e00
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CIIEL' DU DLiP,1RTBNIiNT SC.If:\TIFIQUL' I)GS- .
i:\DRIQUC;S Dt: 1iV3AC I31: U=YIrS S t
. Actj.v,3 Nitrosamines and site of action
--:;spewc3.fic ., A_ ccor.ding to the compound used end according
to therway of application_(intravenous, oral, sub-
c- '~~' ), ~ -_ can , cer is- produced y.n a given, site _such
utane,ous ~~ ,
; as.lung, liver,_ bladder, oesophagus, stomac and
~ Some 60 Nitrosamines - as. well Dialkyl as.more
t complex assymetrical compounds - have been tested.
They are-all-carcinogenic and__what is more organo-
also. for the first time in histouy, in brain.
going into details of what kind of cancer
With
out
.
is produced by which Nitrosamine, it is interesting
to note that-of the symetrical Nitrosamines only
urs.
the Diamyl-compound produces lung tumo
~ _ . ~. -v..
The we1l--known.toxicity of most_ Nitrosamines is not
necessarily parallel_to the,carcinogenic action.
;WMethyl-nitroso-urea=produced kidney tumours,.
~one ca;~e, a unique intrav_enous application of 100 mg
The dosage needed,ta induce cancer in test animals
.is for most_Nitrosamines exceedingly low. In some
instances a unique application_is_sufficient. In
N,i,trosamine..distributes quickly through the
:.T e
11
test,animalTand_is completely metabolised after
24.hours_z(Dimethy-lnitrosamine).
Rats, mice, monkeys, fishes, golden hamsters gave
all similar positive responses. ~ ;
l,t is, thought that the alkylating agents, t]a.ough
alkylati,on=.of__ DNS ~"ACan alter the nucleus and in
such a way be _responsible for the start of cancerous
growth. ~
Page 4: mry24e00
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aRsc-nac:Enr _ADMI\TSTRATEfJR t)IItECTI 412 CT__
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N3`~NNOl -+HCNo
dimethylat' ionu3
se~ms to ta ;e place in many organs._
P , .: _ ;- t-4vp~~
The pres er.ce of,.Methylnitrosamine (lifetime in Ether
at 37°C l sec-.'`- 10-.100 milliseconds in water) in
:the liver.,asw_probable, Formaldehyde has been found
'.,as well as ,CH NFi and HNO The enzymatic.n action of
-the forma.tion,of 7-Me,thylguanine in .the cell. The
Tests.._w*h yC ` ' _,labelled rD,i.met_hylnitrosamine show
,,"~,.- quantity of Methylguanine found in a given organ
:of the test animal._is parallel to the cancer inci-
. ~
;denee.,
The Methylguanine can be found in DNA and also
(less work done) in RNA treated with_methylating
agents.
methy
a
ion).
N-butyl-methy1-nitro amine and tertiary Buty1-methyl-
nitro,samine, both_ c:,l- lab,elled in the Methyl-group,
- show that with__the__.former ---a potent carcinogen - the
activity is in the 7-Methylguanine, and with the
latter no carcinogen - due to steric hindrance,
the activity is in the Guanine and Adenine (no
t
1
FurtherI proof is_brought by Druckrey to show that
"for theqf,ormation of Diazomethane (considered as
theactiv,e agent) a Nitrosamine has to have.two
labil Hydrogen-, atoms.
oxygen is cleaved off to form Diazomethane
,.In the case-,o-f Dimethylnitrosamine, the H of the
K-position is enzymatically replaced by antk3H-group.
In the,sec,ond_rstage, the Hydrogen replacing the one
Methyl-groupraf,terthe_fi^st demethylation + one
Hydrogen from the remaining Methyl-goup + Nitroso-
epro- o o is consis,ts
that _substances_ that
~
, ~
~ ~ . ~. y . -_ J.
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- ~---.~ ~.-
NNZ+HNo2
Page 5: mry24e00
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F:UtRIQUGS DE U\it,\C RGI;\IGS S.\, ..
._T1iL.076 _~7L;0 1
Nr.t:Clt Wi:i P -si:Issr__
not cor,respond to these exigences have nio carcinogenic
action-on,the..teste,d.-animals.
. Active metabolites
Formaldehyde is probably not carcinogenic, HN02
has, according to Druckrey, no carc;nogenic ac-
tion.... Howev-er, Diazomethane-,,is.._a _powerful carci-
._= ., ...
nogeriic agent and a good alkylating agent, too_.
.. . . , .: _ ~
nogenScas a,whole are also mutagenes.
have been tested as_ to,their mutagenic action and
-it could be,shown;,,that all carcinogenic ones are
mutagenes. First results do indicate that carci-
Another.m;inter_e.sti,ng point is that all Nitrosamines
. Inhalation experiments
Nitrosamines.
conditions__l.ung cancers,, hav.e .been_ obtained with
e ie n severa more mon s ung eare nomas ~
will be obtained). Under similar (not identical) I
_.inhalation with tobacco smoke after
:Hamsters _.on..
-
::"88
months show lung lesions (there is reasonable
b Y' f that i 1 th 1 i
The presence_of Nitrosamines in tobacco or smoke
is no.t_=established, nor disproved, for lack of
asuitable analytical method. One method is in
- elaboration_n in,,c-ollabor-ation between Pro.f.
.
Druckrey and labs of the,,German indust,ry.
b) Proposed way of formati.on
The formation of Nitrosamines
`following course :
;
could take
the
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~ ,V3 O r,>Q O{ _ - - ~ ~
Nl.trosopipe~ridine is -a. potent carcinogeri and to ~_
.. , . _
a_lower.degree Nitros-omethylaniline and.Nitr_oso-
~..
*
ty
Alternate possibiliof formation of Nitrosamines
in the lung- ~--~-
_
anabas in,ev.
It is .further believed that the formation of Nitros-
~
amines could take place in the respiratory tract
,due t,o. N2 0 E." ~.. 3 ".. NO + NO and Amines from the lung
tissue.No proof exist3 for this beliei'.
`As a whole one can say that the Nitrosamines are
very"potent carcinogens, potent mutagenes; that
~
-
ganotropic action, that their_effeet_on the chemical
structure of the attacked organism is better known
~
an _ "astonishing relation-betwe-en structure:-and or-
_
they have avery good dose-response relationship
than for most other carcinogens.
-.. ~
`
i
r
It is ev
de
it that this
is only a beginning, sinc_e
man ' oints are et
y p y uncertain, but I feel that this
subject leads to a new way of looking _ at cancer.
-
_
Whatever. ~th e outc.ome might be, we might for on_ce
faced
be one we,witli_a s,omewhat,clearer picture thanthe
ar ., n
e presen y acig.
.'
Have you done work any wor~c on , thesecompounds and; a
sui
table method.for qualitative, or even better
quantitative,analysis ?
- ,
- "
-- -
- -- -
,
-If new developments occur, I shall. ___ keepYyouu in-
formed _
formed and,_I join a bibliography, whichis by no
With kindestre ards _
Sincerely yours,r
-
6
_ _
mean'comRlete, but conveys the more important part.
s -~~- -~ -
Page 7: mry24e00
PETER \\":\-LT7Z
rlr.-so.,nat6PP
ADJII,\IST(LkTGUR-DlRGCTI:UR
Cllrt'DU DI'PARTI:,~llSVT SCU:NTllPIOUT: DES
FADR10UCS DB,T'.\iL\C RfiUNI13S Rr\ ,
Carcinogenic and growth inhibitory activity of
1) F. Dickens and H.E.H. James, Further Study on the
Lactones: _andd related substances. Brit. J. o,f
Cancer, Vo1. XVIII, 1, 100 (1963).
11,. 167 1954).
-2) J.M. Barnes and P.N. Magee, Brit. J. Ind.Med.,
NaturwissenscrhaPaen,_ 4.-, 134 k1901).
5) H. Druckrey, R. Preussmann, D. Schmal and M. MUller,
3) P.N. Magee and T. Hultin, Biochem.J.
4) P.
N. Magee and E. Farber, ibid. 114
,,, . .
_, 5 -57 1g 1
7) Druckre , Schmal and A. Schildbach, Naturwissenschaften,
6) D.W. Fassett, Annual Review of Pharmacology, 3,
289-1g 3
wissenschaf.ten, , 722-23 i961).
9) Druckrey, Preussmann, Schmal and G. Blum, Natur-
10) Druckre , Preussmann and MUller, Naturwisserischaften,
11) Druckrey and Preussmann, Naturwissenschaften, 40,
111 (1962).
- s:, -. _.
12) Druckre and Schma1, Naturwissenchaften, 49, 217
1g 2 ,
13) Druckrey, Preussmann, I. Ackhan and G. B1um,Natur-
-
14) Druckre , Preussmann, Blum and Ivankovic, Naturwiss.,
50, 99 (19637.
T.- ~ . _
wissenschaften,49
451 (1962).
Page 8: mry24e00
PETER NVALT,`/7
dr,sanat,T:PP _ ,
-.ADDIL\iSTRATC(IR-DIRi?cTPUR IiT ClTEF, DU DI:R\P,TiiJT_L:\T_SCIENTIFIpLT_ . DES
FAI3Ri0[i GS DE Ti\EtAC RCU:Q(tis $\
15) ~Dr~uckre and Preussmann, Naturwissensohaften,-4Q,
-
- 21
62
~+
8 (1
r-
9
,
9
nbacco_Science, 3, 139-1 3 1959)
6) R.J. Philippe and E.J. Hackne (Liggett & Myers),
~17) A.J. Haagen-Smit
Science 128, 876 (1958).
~
,
18) A.J. Haagen-Smit, M.F. Brunelle and J. Hara,
A M A A h I d Health 20 399 (1 5o N
) J. Gasparic, Chem. and Ind. 1962 ,43:.
_
-
~
~
