Note: Descriptions are shown in the official language in which they were submitted.
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Use of griseofulvin for inhibiting the
growth of cancers
TECE~NICAL F~ELD
This i,.~.- is a pL~u~ fi~ ~I c--~ -- - that is useful for the ~ of cancers
and tumors, p=-l;r~ in h~unan and warm blooded animals. The f~ --"- contains
-~ h~. It can be used in A ~ with other c' --- -'~~~q~ ~ agents also.
S BACKGROU~ID OF T~E INVENTION
Cancers, i~ ;nE '~ ' ~ are the leading cause of death in animals and hurnans. The
exact cause of l~ mi~ is not known, but links between certain activities such as smoking or
c,~s u~ to w~_h-og_-~ and the ir~ nce of certain types of I ' - and turnors has been shown
by a number of l~e~.h~.a.
Many types of ,1...... ~ ;c agents have been shown to be effective againsat cancers,
tumors and I ' but not all types of cancer and tumor cells respond to these agents.
u"ru~ , many of these agents also destroy normal oells. The exact ~ for the
action of these c' , - agents are not always known.
Despite a.l~ l~.,S in the field of canoer and l~ul~m~ n~.lta the leading t~ , ~ to
lS date are r~ ti- n and ~1- -----11-- ~1~ and bone marrow ~ 'h. ..--~ll.. .a~ d~
are said to fight canoers that a~e p~uh.ukul~ a~.e~;~_. Such cytocidal or ~jt~at~li- agents work
best on cancers with largc growth factors, i.e., ones whose oells are rapidly dividing. To date,
1~."~ ~A~ in p~uh-,~l~ estrogen. ~ ~uO_.t~,.une and i une, and some ~ ~,-ulu~l by
a variety of ~ lahl.g agents, and anti ~- I"hnl;~ form the bulk of i ~ available
20 to ~ Ideally ~ ~ - agents that have ~ ;r..:/~ for '~ ~ ~, canoer and tumor oells
while not affecting normal oells would be ~ el~ ~If-' -- ~~ U-lrU-i ' ~.y, none have been
found and instead agents which target especially rapidly dividing oells (both diseased and normal)
have been used.
Clearly, the d~ lo~..." .~ Of . .t~ lc that would target canoer or le -l-~mi:- oells due to
2~ some unique ~ r.;~ for them would be a b.cdkllu. _- Alt~..~h._l~, -'- that were
.,~tolu~c to It ~ ~ or cancer cells while exerting mild effects on normal oells would be
~ ~ '- Therefore, ill is an object of this i,.~_.,tion to provide a ph-..--'- ~j..l;~ ~1 CC~ that
is effective in treating '~ ' ~ with mild or no effects on normal blood oells
More !q~ifi~lly, it is an object of this invention to provide a ~u--~ u~ g a
30 ph ~- ' carrier and a o~ia~,~r~ i" as defined herein along with a method for treating
cancer, IP-.1,Pm;~ and tumors.
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The u e of ~ c~rulvin in co~ l;n ~ with other ch~mr~th~ ;c agents which are
effective in dc~lluyingth~e tumor is a novel method of tl~at~ nl Griseofulvin can also be used to
treat viral infi~cti-~nc in the presence of a p
SUMMARY OF THE INVENTION
S A ph~ ti~1 co~ ion for tll,atl~ -t of m~mm~llc, and in pa li-,ular, warmblooded animals and humuns, which are affected by I ' - Cc)--~ g a ph~ i carrierand an effective amount of ~;.i~vrul~in Griseofulvin has the formula:
CH30 ~ ~ ~
These ,~ c can be u ed to inhibit the growth of le~ r~;o turnors and cancer
oells in hurnans or anim~ls by ~ t ~~;o~ of an effective amount either orally, rectally,
topically or pa~ 11y, or illtla~ ,. These ~c l n~ do not ~ ri ~ ~11y a~fect hc-althy
cells.
Pl~ can also be used in co- I~ with ~j.iacvr,llvin as can l~.. lh
lS agents.
DEl'AILED DESCRIPTION OF T~E INVENTION
DE~INlTIONS:
As used herein, the term "co l ;~;~g" means various r~r can be ~ 1, 1~,
in the pt~ of this invention Accul li~ly, the terrns "co~ g
20 , - 'Iy of n and ~c g Or are ~ d in the term ~ g
As used herein, a "~ " ~ 11y ~ u c~ is one that is suitable for use
with humans and~or animals without undue adverse side effects (such as toxicity, ilTit~tinn, and
allergic response) co-- . ..~ te with a ~._ ' '- benefit/risk ratio
As used herein, the term "safe and effective amount" refers to the quantity of a c~ ~''~ -.1
25 which is ~ r: .-t to yield a desired thf ~ ;c response without undue adverse side effects (wch
as toxicity, ~ t~ n~ or allergic response) L('''~ with a ,~ )r ' I~ ~n-,r~ ;ak ratio when
used in the rnanner of this i..~f.lliO... The specific "safe and effective amount" will, ~.liou~ly,
vary with such factors as the particular co~i;l;o - being treated, the physical c~ l;l;o-- of the
patient, the type of mamlr~ being treated, the duration of the lltiall..c.ll, the nature of co..,...,,_nl
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--3--
therapy (if any), and the specific r~)". .I~I;nnc ~ .i and the structure of the CO"~1~UY~ or its
d..iv~ti~
As used herein, a -~ t;~ -' addition salts" is salt of the anti ll .,L. - ~ C~..~l~....A
with an organic or hlOI~,diUC acid. These ~.ef...e;i acid addition salts are rhlnnA~oc, blu..~idc,s,
S sulfates, nitrates, ph~y~ t. -, R~C ' S, finrmzlt~5~ tartrates, mzll ' , malates, citrates, ~ u ~h ~,
salicylates, ~u-I,at. " and the like.
As used herein, a "~h~.~ e-~ l carrier" is a ph~ ly a "~ solvent,
r ,, agent or vehicle for delivering the anti '~ ' ' agent to the animal or human. The
carrier may be liquid or solid and is selected with the planned manner of -d~ U"~ A in mind.
As used herein, "cancer" or "l ~ ~ " refers to all types of cancers or - ,' or
...~1;~.- -- ~ disease which attack normal healthy blood cells or bone marrow which produces blood
cells which are found in ~., ~
As used herein, "viruses" includes viruses which cause diseases in war n blooded animals
e HIV, i"n.,. ~-,~ ~hil~uvilu~c;~, herpes and the like.
As used herein, "g-i~ûrljlvin" means 7-Chloro-2',4,6-L-i.. _ll-u.~-6 ._lh,~ JilU
rt ~,..r~ 2-(3H),1'-[2~ lolt~ ] 3,4'~ione. It is an ~ .uducoi by
p . 7r griseofu~vum
As used herein "F~ ~ are ~ ' '- such as l.;l..ulidi-le and its cis-isomer which
are used in c~ -- with ~ ~rulvi... r.. can suppress the immune system or
20 enhance the c~li~_"_~ of the drugs.
As used herein ~ nlh ~ ;c agents" includes DNA-' '._ Agents, Antime-
tabolites, Tubulin-T 'v~ Agents, ITn..~ O~ agents and others, such as ~p--~gi~ -~ or
h~.Lu~y~ ~.
B. GRISEOFULVIN
~ YJr~lViU~ has the following ~Uu.luc.
C H30
It is prepared according to the method Af ~ . ;I~A in U.S. 3,069,328 issued to TTn. L. ..
(1962) and U.S. 3,069,328 issued to Dorey et al. (1962)
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C CUEM OT~ERAPEUTIC AGENTS
The d~r~ ir agents are generally grouped a_ DNA~ Agents,
A~ ;t~ C, Tubulin-Interactive Agents, IIo- I agents and others such as A.~ ~ or
L r~UA~r~U~. Each of the groups of ~ agents can be further divided by type of
5 activity or co--.l~--u~l The rh~moth.._l,~ agents u_ed in c~ ;o~1 with 2~i~oîulvin
include ~-- ~ ~l~f ~-, of all of these groups. For a detailed ~ of the ~ ...n~ f .~;r agents
and their method of ad~ ul;on, see Dorr, et al, Cancer ~ th~rapy Hondboo~c, 2d edition,
pages lS-34, Appleton rl~ Lange (C ~ , 1994) herein i..~.~,u.,~t~ by l~f~"~ c.
DNA-T '~,o Agents include the aL~cylating agents, e.g. Cicp1~'in C~,rc1r~l~hn~
10 All.1 ~, the DNA strand-bl~kà6_ agent_, _uch as Bl~...~, the ih.t~lr~lil.g
1O~ c.J.~ ~; ;e II inhibitors, e.g., Dactinomycin and Dw~u-l ' ' ); the r ' ~lating
"~r;~ e II inhihi~rlrc such as, F~posi~1r and Tc~ ~s~i~, and the DNA minor groove binder
Plc~ll., li...
The alkylating agents form covalent chrmir~l adducts with cellular DNA, RNA, andlS protein ---1( ' - and with smaller amino acids, gh~ hin~ and similar rhrmir~l~ Generally,
these allylating agents react with a .~ ~ph;1ir atom in a oellular co~l ;n -~ such as an amino,
carboxyl, r~ n.y,~l group in nucleic acids, proteins, amino acids, or g' ~-~' ~ - T_e
~ ' ~ and the role of these alkylating agents in cancer therapy is not well u~ ~.r~r~OA
Typical aL~l,~ing agents indude:
Nitrogen mustards, such as Ch1o1~ u~ C~lophr~ T " ~,
Mechlo..l1--.--;1~ M~ h~ Uracil mustard;
AziAdine such as Thiotepa
t; ~~ ~ esters such as ~ llf~n
nitrosou~5,suchasC~.,... l ;..r Tr ~,S~ l~t~
F ' c , ' , such as ~i, ' ' GUIJOplf~lil-,
' ' r~l~.,li~_ alcylator, such as MilOl--yr i.-, and Plu~l,a~il.e, D~-~l,~ne andAlll.
DNA strand breaking agents include Bleollly~
DNA l~ nhih;tor~ include the following:
T ~ ' , suchas A.--~ e Da~;~inoll.. ~ , Daunorubicin, Do~o. ' ' '
Idall ' ' ~ and M;l.,._u~ulle.,
- ' ~lators, such as F~ f and Te-~ 1c
The DNA minor groove binder is Plic~..~_h-.
The ~ l~l;t~, interfere with the ~lU hJ.,Iion of nucleic acids by one or the other of
3S t~o major ",~h~ Some of the drugs inhibit l,lu~lu~lioll of the deo~y-i~
lA~ t'_ that are the il ---" Pl~ U~ for DNA synthesis, thus inhihiting DNA
Some of the eu~ are S--n~ ly like purines or pynmi~ oS to be able to
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-5-
for them in the anabolic '- ' patl~ s These analogs ean then be ~ ~
into the DNA and RNA instead of their normal cu~ t-.~alLs. The ~ oi;t~ ~ useful herein
inelude:
folate ~ such as M_lholl. and l-i...~,l..
S pyrimidine ~ l~o ~ " sueh as Fluo-uu c~ -il, Fluo.udco,~idine, CB3717, A7qr~ n
Cy~;~.e, and Floxuridine
purine ~ ~g. ;~1~ inelude M~.~ , . 6-Tl~ ;.f ~-J~hi.~,rl ;
wgar modified analogs inelude C~_~dl,i-.c, Fl ~ h;
'- ' ' .~1U~,LO- 1 ~ inelude h~u~a
Tubulin T ~_ agents act by binding to specific sites on tubulin, a protein that
pol~ .i~s to form cellular ~. i_-.hL '- Mi,-ulLL ' are eritieal eell structure units When
the ~ ~,_ agents bind on the protein, the cell can not form l~iwulub~lC5 Tubulin I"t~.~,li,_
agents include Vi~ and V ~'- both q-l~ql- '' and Pa~,lila.~,l.
~Tnnntlnql agents are also useful in the l,~aL,...,.II of eancers and tumors. They are used in
15 ~ lly - ~-r ~ tuunors and are usually derived from natural sources. These include:
~,truO_.~" . ;_~ ~ e~,l.u~, and Ethinyl ~ct~tliol snd Diethylcti~
~hl." ~ GP.. and T<~ d~
I,,~,_;,lh,s sueh as II~uA~ ,~t~,.une caproate, Med-u.~ ,u~tu.unc, and Mct,_~tlul;
ai~uO_.~ sueh as l~;.lua~.u~ ui~c ~ r~ ~, nl~ui~yl--_:~t~.-O~C,
I--~lI-ylt~tual~-u--_,
Adrenal co.li,u~ ' are derived from natural adrenal eortisol or hydlu,OIliaOll.,. They
are us~d beeause of their anti " y benefits as well as the ability of some to inhibit mitotie
liv -- and to halt ~NA ~ -' These enmro~ c inelude,
M_~l~ , and P-~
2S T~ e hormone ~~,1~;. o hormone agents or O ~'~ u~ih~-relea ing hormone
are used primarily the l.~ ~ of prostate cancer These include leuprolide acetateand ~-~,li., acetate. They prevent the ~ , IllL~,aia of steroidâ in the testes
.~nti~ ~' antigens include:
~ti~ u~, ni~, agents such as Tz~Tn~cifi~ n
' ug~,.. agents _uch as Flutamide; and
allLiadl~aal agents such as l~litotqnt~ and ~. ;- .~1"~1;. -i 1c
E~d-UA.~-W appear_ to act primarily through inhibit ~r of the enyme .;1~ liA~.
~ is an enyme which converts -~ t~ to rU~ ~r ~ ~ it~n~l aspartic acid and
3S thus blocks protein a.~lllL-,~;s in the tumor
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D. POTENTIATORS
The "~ can be any material which i.l.~.lu.es or increase the efficacy of the
ph~ c.--~ or aets as an ;~ ~ o~ p~,~3r. One such F- is triprolidine
and its cis-isomer which are used in cu~ inn with the ,~ ;c agents and t~e
S ~i~orUlvi.,. Triprolidine is de~li~ in US 5,114,951 (1992). Another p is
pr.-~ '~, lII Rr..,;~ 2_~11, ~ acid; [B-(2-ben7imiA ~-) p.l, acid; 2-(2-
ca lu~ .,lllyl)brn7imi~ ~ propa_oll. Pl- ~ 'e is a non-specific active i,.. m ,~
agent against viral and bacterial infi~ti - - - and can be used with the co- ~1 n~ :~ io.~ claimed herein.
It is effective with gli~eorulvi-l alone in treating cancers, tumors, le.~ mi~ and viral jj.F~ or
10 cv~ ~ with ~ JI~ nll~ V~;C agents.
P~o~ - acid and its salts and esters can also be used in ~ ;"n with the
r'~ 1 C~ IU)S;I;OnC claimed herein.
~ n~inYi,l~nr vitamins such as vitamins A, C and E and beta-carotene can be added to
these cc.--~, oc;~
15 E. DOSAGE
Any suitable dosage may be given in the method of the i.,~ r The type of e ~
and the carrier and the amount will vary widely dr~nAing on the species of the warm blooded
animal or human, body weight, and the type of cancer or tu~nor or viral infiection being treated.
Generally a dosage of between about 1 milli~m (mg) per kilogr~m (Icg) of body weight and about
20 8000 mg per kg of body weight is suitable for either the griseofulvin or the _h - , ~
agent. Plef~,.~l~ from 15 mg to about 5000 mg/kg of body weight is used. Generally, the dosage
in man is lower than for small warm blooded m~mm~lc such as mice. A dosage unit may
co ~1" ;~ a single ~ 1~ .A or mixtures thereof with other ,O...In, ~~lc or other cancer inh;t~iti~g
~ The dosage unit can also _r diluents, ~ d~ carriers, l;l u~-~r c and the
2S like. The unit may be in solid or gel fonn such as pills, tablets, capsules and the like or in liquid
for n suitable for oral, rectal, topical, hlLI~.~ e.~JuS injection or pal~ r or
injection into or around the bone marrow. The range and ratio of griseofulvin tochr ~ , ..t;r agent will depend on the type of cancer or tumor being treated and the p~ Li~,ul~
l~hr nntk .;~ agent.
30 F. DOSAGE DELIVERY FORMS
The ~h..,lh.,~ ;,, agents, griseofulvin and, optionally, the put~ are typically
mi7~ed with a pl.~ lly ~ carrier. This carrier can be a solid or liquid and the
type is generally chosen based on the type of a-l~ ion being used. The active agent can be
C~ ~ 't~Cd in the form of a tablet or capsule, li~cnnlr, as an a~l~ t~Yi powder or in a
3S liquid form. F ~ r of suitable solid carriers include lactose, sucrose, gelatin and agar.
Capsule or tablets can be easily formulated and can be made easy to swallow or chew; other solid
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_7_
forms indude granules, and bulk powders. Tablets may contain suitable binders, l~
diluents, ~' t~o ,, agents, coloring agents, navo~ g agents, flow-inducing agents, and
melting agents. F - '-- of suitable liquid dosage forrns include ~ c or ~ in
water, pha~ lly ~ ~.p! ' ~~ fats and oils, alcohols or other org nic solvents,; ~ e
5 esters, ~ c syrups or elixirs, ~ )nc, solutions and/or ~ - lr -~:o~c ~ t ~l from
non-effervescent granules and ~,ff...~.~ prepA-r~ti~nc ,- COh~ A, from ~f~ l granules.
Such liquid dosage forms may contain, for, . 'e suitable solvents, yl~,~.vali~5, e..~ if~i-.O
agents, ~ ne agents, diluents, _.._ th~ PnPr~ and melting agents Oral dosage
forms optionally contain na~v.~lt~ and coloring agents. Pal~ ~ ' and inlla~ u~ forms would
10 also include minerals and other m~Pri~lc to make them ~- It with the type of injection or
delivery system chosen.
Specific ~ of pha. .-- ~ ;. al a ,ep~ ' 'o carriers and, ~~ ~ ~ that may be used
to r_ ' ~,~ oral dosage forms of the present il.~ ioll are d~ A in U. S. Pat. No. 3,903,297
to Robert, issued Sept. 2, 1975.Tcchni~lu~c and ~ ~po~;~;o~ for making dosage forms usefill in
lS the present invention are dr~-;I~d in the follo ving ~ e~. 7 Modern Pl-a. ~ ~ ul;- 5. Chapters
9 and 10 (13anker ~ Rhodes, Editors, 1979);T l~-~--al~ et al., Pl-a~ _l Dosa~ee Forms:
(1981); and Ansel, Il-llu lu-,lio-- to Pl.~ ...~ al Dosa e Forms 2nd Edition (1976).
G METHOD OF TREATMENT
The method of L.~ can be any suitable method which is effective in the ll~ of
20 the ~_ ' cancer or tumor type being treated. Treatment may be oral, rectal, topical,
pa~e ' or i.~L...~_nvus ~I...;ni~u.-~ or by injection into the tumor or cancer. The method of
applying an effective amount also varies d- ~ n-l;ug on the le~ Pmi-- cancer, tumor or virus being
treated. It is believed that ~ ? ~l---~,..l by hllla~-~luus, 5uh~ n~ , or ;~
a~ of the ~;~rul~ill, r~ d with an a~.. upl;alt: carrier, a~ ' canoer ~ ' ' g
25 ~ . ' or ~ , ' or diluent to facilitate ~p~ on will be the l l~,fe,l~ method of
~ ~ ~ ~ ~ ~ th~ ' to warm blooded animals.
In addition to the use of ~hemnl~ agents and p ~isevr~ ill can be
~ ~ ~ ' with fi-ur;~ Jr;, L~ or other antiviral agents. P~ l h.,.~ - ~ and
r.... .nF"~ :d ~ ~ include call~ ,; " " n U~o~ e, benomyl, ~;ly~hv~tu and pl-r c - ' 'e
Example 1
In an acute HIV in vitro model, griseofulvin inhihi~Pd viral llr~ ~ by 98% at
lOIIg/ml with a Ih .~ C index of 5.3. AZT, a known ~V drug, also inhihi~Pd viral l~
by 98% at lllg/ml with a ~h~ ;c index of 12,500. The Ih~,.a~ lic index is the ratio of toxic
dose of drug to ~ ll~. ~- :-.--c dose of drug.
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Example 2
In an in vivo mouse study for I ' '- (P388), ~.i~orulvin showed an increas~ in the
survival time relative to a non treated control of 156% at 4000 mg/kg dose; 188% at 5000 mglkg
dose; and 218% at 6000 mgAcg dose.
S Example 3
In an in vivo mouse study for, ~ (B16), ~fi~r~vi., showed an increase in the
survival time relative to a nU~ control of 165% at 4000 mg/kg dose; 179% at S000 mg/kg
dose; and 201% at 6000 mglkg dose. Cytoxan at 300 mg/kg showed an h.w~d survival rate of
192%.
Example 4
In an in vitro ~m..g for P~ , type A-l, cell line WI-38, g-;~rulvin was
effective at 100 llg/ml. The positive control was A-36683 of Abbot Company, (S,S)-1,2-bis(S-
methoxy-2~ --~yl)-1,2~~ J ol A-36683 has a ~ Jlir index of 1000-3200.
fi~l~,in has a i' , - ~- index of 1-2. (See S~hl~irhPr et al, Applied Microbiolo~y, ~, No.
15 1, 113-116 (1972).
Example 5
Solid tumors removed by patients are minced into 2 to 5 mm r -~ ~-1~ and ~
placed in McCoy's Medium 5A plus 10~/O heat hla~ newborn calf serum plus 1%
r llin/~ . Within 4 hours, these solid tumors are ,..f~ ly ,i;~ t-'~ with
20 scissors, forced through No. 100 stainless steel mesh through 25 gauge needles, and then washed
with McCoy's medium as dc~cfil,cd above. Ascitic, pleural, p~ fi~ lial fluids and bone marrow
are obtained by standard ~ c The fluid or marrow is placed in sterile co- ~
' ~ - g 10 units of ~/lc~va~ e free heparin per ml. of m~ gJl~nt fluid or marrow. After
~ ~~- '~ue~- ~ - at 150 x g for 10 minutes, the cells are haJ ~ d and washed with McCoy's medium
25 plus 10% heat h la~ calf serum. The viability of oell ~ ,c:.~c is J~ ~- ~---;---'A on a
h~ te~ with trypan blue.
Cells to be cloned are 5 ~ ~ in 0.3~/O agar in enriched CMRL1066 _,"'~ ' with
lS~/. heat in~li..~ ~ horse serum, pPnirillin {100 units/ml), ~ (2mg/ml), g' ~ ~ -
(2mM), insulin (3 units/ml), ~ -e (0.6 mg/ml), and HEPES buffer (2mM). For the
30 ~ ~ exposure test each ~u~ is added to the above mixlure. Cells are placed in 35
mm petri dishes in a top layer of agar over an ullde~ . of agar to prevent growth of r.b.~~
Three plates are prepared for each data point. The plates are placed in a 37~C inr ~ - r, and are
removed on day 14 for counting of the number of colonies in each plate. The number of colonies
(defined as 50 cells) formed in the 3 r~ treated plates is Cw~uc;i to the number of~S colonies fonned in the 3 control plates, and the percent colonies S~ 'ViVil-g at the cc ~ - of
' can be; ~ ' Three positive control plates are used to ~i- t~ i-e s~ival rate.
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g
O.i' ~ s ' ~, - ' at 200 ~g/ml is used as the positive control. If there is <30% colonies in
the positive control when cu~ d to the u..l.~t~.d control, the test is evaluated.
At con~ of 0.5 and S.0 llg/ml in a single dose e,~ ~oru~ was not
effective (0/1) against tumors in this test. At c~ of S0.0 llg/ml in a; exposure
S ~ ~ ~i~rul~ill was effective against colon, lun& n~ 1 cell, and ovarian cancers .
Over all S of 6 had SS0% survival.
