t19
`United States Patent
`4,885,171
`[11] Patent Number:
`Surendraetal.
`
`[45] Date of Patent: Dec. 5, 1989
`
`[54]
`
`[75]
`
`[73]
`
`[21]
`
`[22]
`
`[63]
`
`USE OF RAPAMYCIN IN TREATMENT OF
`CERTAIN TUMORS
`
`[52] U.S. Cl. ceessessscscccosescenessssssscsssssrsssesseers 424/122
`[58] Field of Search 00.0... esssssseesesessesssees 424/122
`
`Inventors: Sehgal N. Surendra, Doliard Des
`Ormeaux; Claude Vezina, Oka, both
`of Canada
`
`[56]
`
`References Cited
`PUBLICATIONS
`
`Assignee: American Home Products
`Corporation, New York, N.Y.
`
`Appl. No.: 592,193
`
`Filed:
`
`Mar. 22, 1984
`
`Endicott, J. of the National Cancer Institute, The Che-
`motherapy Program,vol. 19, No. 2 (20th Anniversary)
`Aug. 1957, pp. 275-293.
`
`Primary Examiner—Jerome D. Goldberg
`Attorney, Agent, or Firm—Walter Patton
`
`Related U.S. Application Data
`Continuation of Ser. No. 126,276, Mar. 3, 1980, aban-
`doned, which is a continuation of Ser. No. 957,626,
`Nov.3, 1978, abandoned.
`
`ABSTRACT
`[57]
`Methodsfor using rapamycin in the treatmentof certain
`cancers or tumors are disclosed.
`
`[51]
`
`Tint. C14 sec essessssessescsssscessssensscessnnsees A61K 35/74
`
`7 Claims, No Drawings
`
`West-Ward Pharm.
`Exhibit 1042
`Page 001
`
`West-Ward Pharm.
`Exhibit 1042
`Page 001
`
`

`

`1
`
`4,885,171
`
`2
`schedule of dosing can range from one to five times per
`day to a single dose given every two to ten days. Such
`dosages and scheduling of administration must be deter-
`mined on an individual basis, depending upon the tumor
`or cancer, nutritional state of the mammal, age of the
`mammal, toxicity in each individual, etc.
`Rapamycin reduces tumorsize in and prolongs the
`survival time of tumor-bearing mammals. More specifi-
`cally, rapamycin is useful for controlling the following
`carcinogenic tumors in a mammal: lymphatic leukemia,
`colon, mammary, melanocarcinoma and ependymoblas-
`toma. The effectiveness of rapamycin in this respect can
`be demonstrated in the laboratory with rodents having
`transplanted tumors. Details of methods used to evalu-
`ate this effect are described in various publications; for
`example, R. I. Geran et al., Cancer Chemother. Rep.,
`Part 3, 3, (No.2) 1-103 (1972) and referencestherein. In
`addition the protocols for the antitumortests are avail-
`able from the National Cancer Institute, Bethesda, Md.,
`U.S.A.
`Tables 1 to 6 show the effects of therapy with rapa-
`mycin on various tumoror cancers in rodents.
`Morespecifically, Table 1 shows the prolongation of
`survival time of female CDF; mice implanted with
`lymphatic leukemia P338 by administering rapamycin;
`Table 2 showsthe reductionin size of colon 38 tumors
`in female BDF; mice by administering rapamycin;
`Table 3 showsthe prolongation of survival time of male
`CDF; mice implanted with colon 26 tumors by adminis-
`tering rapamycin; Table 4 shows the reductionin size of
`CD8F; mammary tumors in male CD8F1 rats by ad-
`ministering rapamycin; Table 5 showsthe prolongation
`of survival time of female BDFmice implanted with
`B16 melonocarcinoma by administering rapamycin; and
`Table 6 showsthe prolongation of survival time of male
`Swiss mice implanted with ependymoblastoma by ad-
`ministering rapamycin.
`TABLE1
`
`
`USE OF RAPAMYCIN IN TREATMENT OF
`CERTAIN TUMORS
`
`This is a continuation of application Ser. No. 126,276,
`filed Mar. 3, 1980 which in turn is a continuation of
`application Ser. No. 957,626, filed Nov. 3, 1978, both
`now abandoned.
`
`5
`
`BACKGROUND OF THE INVENTION
`1. Field of the Invention
`This invention relates to the use of rapamycin as an
`anti-cancer or anti-tumor agent.
`2. Description of the Prior Art
`Rapamycin is an antifungal antibiotic described by C.
`Vezina etal., J. Antibiot., 28, 721 (1975), S. N. Sehgal et
`al., J. Antibiot., 28, 727 (1975) and S. N. Sehgal etal.,
`U.S.Pat. No. 3,929,992, issued Dec. 30, 1975,filed Apr.
`12, 1974. Rapamycin is extracted from a streptomycete
`(Streptomyces hygroscopicus) isolated from an Easter
`Island soil sample andis particularly effective against
`Candida albicans both in vitro and in vivo.
`In addition, a recent report by R. R. Martel et al.,
`Can. J. Physiol., 55, 48 (1977) describes the use of rapa-
`mycin for the prevention of the development of two
`experimental
`immunopathies
`[(experimental allergic
`encephalomyelitis (EAE) and adjuvantarthritis (AA)].
`The latter report also describes the inhibitory effect of
`rapamycin on the formation of humoral (IgE-like) anti-
`body. This report concludes that immunosuppressant
`activity of rapamycin appearsto be related to inhibition
`of the lymphatic system.
`SUMMARYOF THE INVENTION
`
`According to this invention a methodis provided for
`treating carcinogenic tumors in a mammal which com-
`prises administering to the mammal an antitumoreffec-
`tive amount of rapamycin. More specifically, rapamy-
`cin reduces tumorsize in and prolongsthe survival time
`of tumor bearing mammals.
`
`DETAILS OF THE INVENTION
`
`20
`
`25
`
`According to the present method, rapamycin is em-
`ployed as the active agent. Theisolation and description
`of rapamycin is given in U.S. Pat. No. 3,929,992, cited
`above, herein incorporated by reference.
`Rapamycin is administered to a carcinogenic tumor
`bearing mammal for the purpose of reducing the tumor
`size and prolonging the survival time of the tumorbear-
`ing mammal, either orally or parenterally.
`While rapamycin can be administered above, e.g. as a
`sole component of a filled capsule, it is preferred to
`formulate the compound in various dosage forms for
`oral or parenteral administration, e.g. tablets or sterile
`solutions. Such formulations are described in U.S. Pat.
`No. 3,929,992, cited above.
`When utilizing rapamycin for the treatment of tu-
`mors, the total dose of active agent can range from 0.5
`to 500 mg per kg of body weight per day with a pre-
`ferred dosage range from 10 to 250 mg per kg of body
`weight per day. Howeveras the dosage of rapamycin to
`be administered by the method of this invention will of
`course vary with the tumor or cancer and tolerance of
`the mammal,it is preferred to initiate treatment of the
`tumor bearing mammal with a low daily dose of rapa-
`mycin and then to gradually increase the dosage until a
`desirable reduction in tumorsize is achieved without
`causing any harmful or deleterious side effects. The
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Effect of Rapamycin on Survival Time CDF; Mice
`Implanted with Lymphatic Leukemia P-388(ascetic).
`Ave. Wt. Difference
`MST
`of Animals
`days
`(T-C, g)
`-1.9
`~2.4
`—1.6
`~19
`-1.6
`—0.6
`
`T
`14.1
`13.1
`13.7
`14.3
`13.9
`13.9
`
`Cc
`10.2
`10.2
`10.2
`10.2
`10.2
`10.2
`
`Survivors
`on Day §
`6/6
`6/6
`6/6
`6/6
`6/6
`6/6
`
`Dose/Inj
`mg/kg
`400
`200
`100
`50
`25
`12.5
`Treatment:
`Nine intraperitoneal injections starting on day one in a vehicle of saline with
`Tween-80 (Trade Mark for a derivative of Z-sorbitan mono-9-octadecenoate poly-
`(oxy-1,2-ethanediy])].
`Evaluation:
`T/C % = Median Survival Time (MST) in days of treated animals (T) controi
`animals (c) x 100. A T/C % of 125 or greater is considered as a significant proton-
`gation of host survival. Evaluation done on day 30.
`
`T/C %
`MST
`138
`128
`134
`140
`136
`136
`
`TABLE2rt
`
`Effect of Rapamycin on Colon 38 Tumor Weight in Mice
`Ave. Net Wt.
`Difference
`of Animals
`(T-C, g)
`—34
`—2.0
`—0.8
`—0.8
`—0.4
`
`Dose/Inj
`mg/kg
`400
`200
`100
`50
`25
`
`Survivors
`Day 5
`10/10
`10/10
`10/10
`9/10
`10/10
`
`MTW
`mg
`
`T
`188
`209
`272
`320
`368
`
`Cc
`810
`810
`810
`810
`810
`
`T/C %
`MTW
`23
`25
`33
`39
`45
`
`West-Ward Pharm.
`Exhibit 1042
`Page 002
`
`West-Ward Pharm.
`Exhibit 1042
`Page 002
`
`

`

`TABLE2-continued
`
`4,885,171
`
`4
`TABLE6
`
`Effect of Rapamycin on Colon 38 Tumor Weight in Mice
`Ave. Net Wt.
`Difference
`of Animals
`(T-C, g)
`0.4
`
`MTW
`mg
`Cc
`810
`
`T
`368
`
`T/C %
`MTW
`45
`
`Survivors
`Day 5
`10/10
`
`Dose/Inj
`mg/kg
`12.5
`Treatment:
`Single intraperitoneal injection on days 2, 9 and 16 in a vehicle of saline with 10
`Tween-80.
`Evaluation:
`T/C % = Median tumor weight (ATW) estimated from tumordiameteroftreated
`animals (T)/control animals (C) x 100. A T/C % of 42 orless is considered as a
`significant inhibitor of tomor growth. Evaluation done on day 20.
`
`Effect of Rapamycin on Ependymoblastoma in Swiss Mice
`MST
`Ave. Wt. Difference
`Survivors
`of Animal
`days
`on day 5
`(T-C, g)
`—3.3
`10/10
`—2.2
`10/10
`—1.3
`9/10
`—2.0
`10/10
`-1.0
`10/10
`
`T
`44.0
`26.0
`34.0
`34.0
`32.3
`
`c
`18.1
`18.1
`18.1
`18.1
`18.1
`
`Dose/Inj.
`mg/kg
`200
`100
`50
`25
`12.5
`Treatment:
`Single intraperitoneal injection on each of days 1 through 9 in a vehicle ofsaline
`with Tween-80.
`Evaluation:
`T/C % = Median Survival Time (MST) in daysof treated animals (T) control
`animals (C) x 100. A T/C % of 125 or greater is considered as a significant
`prolongation of host survival. Evaluation done on day 60.
`
`T/C %
`MST
`243
`143
`187
`187
`178
`
`5
`
`15
`
`TABLE3.
`
`
`Rapamycin also can be used to produce beneficial
`effects in the treatment of malignant tumors when com-
`bined with a therapeutically effective amount of an
`antineoplastic agent commonly used in cancer therapy.
`Such antineoplastic agents
`include the alkylating
`agents, for example, busulfan, chlorambucil, cyclophos-
`phamide, mechlorethamine hydrochloride, melphalan,
`pipobroman, thiotepa and uracil mustard; antimetabo-
`137
`lites, for example, cytarabine, fluorouracil, floxuridine,
`135
`mercaptopurine, methotrexate and thioguanine; miscel-
`151
`ie 5 laneous anticancer agents, for example, dacarbazine,
`hydroxyurea, mitotane, procarbazine hydrochloride,
`quinacrine hydrochloride, vinblastine sulfate and vin-
`cristine sulfate; estrogens, for example, chlorotriani-
`sene, conjugate estrogens (e.g. PREMARIN @®), dieth-
`ylstilbestrol and the like; androgens, for example, meth-
`yltestosterone, testosterone and the like; adrenal corti-
`costeroids, for example, prednisone and thelike; proges-
`tagens, for example, megestrol, hydroxyprogesterone
`caproate andthe like; radioactive isotopes; and antibiot-
`ics, for example, bleomycin sulfate, doxorubicin hydro-
`TABLE 4
`
`“WithuOfRananvGhOnCDIFMaoe45 chloride and the like. Suitable methods of administra-—EffectofRapamycinonCD8F;MammaryTumorsinCD8F)Rats_35tion, compositions and dosages of the antineoplastic
`
`Ave. Net Wt.
`.
`agents are described in medical textbooks; for instance,
`_
`Difference
`;
`MTW
`“PHYSICIANS’ DESK REFERENCE”, 32nd ed.,
`Dose/Inj Survivors _mg____1/C%—Medical Economics Co., Oradell, N.J. U.S.A., 1978 andof Animals
`
`
`(T-C,g)
`Day5
`Cc
`“AMA DRUG EVALUATIONS”,3 ed. PSG Publish-
`mg/kg
`T
`_MIW
`400
`—6.6
`4/10
`0
`3200
`—
`40 ing Company,
`Inc., Littleton, Mass. U.S.A., pp
`200
`~6.5
`10/10
`323
`3200
`10
`1106-1151, 1977. When used in combination, rapamycin
`100
`—4.8
`10/10
`448
`3200
`14
`is administered as described previously; however, a
`50
`—41
`10/10
`755
`3200
`23
`lower dose can be used for efficacious results.
`25
`—2.4
`10/10
`825
`3200
`25
`Weclaim:
`12.5
`—0.8
`10/10
`928
`3200
`1. A method of treating transplanted tumors in a
`Treatment:
`45 transplanted tumor bearing mammal, wherein said
`Single intraperitoneal injection on days 1, 8, 15, 22 and 29 in vehicleofsaline with
`Tween-80.
`tumoris selected from lymphatic leukemia, colon, mam-
`Evaluation:
`mary, melanocarcinoma and ependymoblastoma tumors
`which comprises administering to said mammal ananti-
`T/C % = Median tumor weight (MTW) estimated from tumor diameter oftreated
`animals (T)/control animals (C) x 100. A T/C% of42 orless is considered as a
`tumoreffective amount of rapamycin.
`significant inhibitor of growth. Evaluation done on day 30.
`2. The method of claim 1 wherein rapamycin is ad-
`ministered orally or parenteraily.
`3. The method of claim 2 wherein rapamycin is ad-
`TABLE5tene
`ministered intraperitoneaily as a solution in saline with a
`Effect of Rapamycin on B 16 melanocarcinomain BDF] Mice
`derivative of (Z)-sorbitan mono-9-octadecenoate poly-
`Ave. Wt. Difference
`MST
`(oxy-1,2-ethanediyl).
`Survivors
`of Animals
`days
`4. The method of claim 2 wherein rapamycin is ad-
`on Day 5
`(T-C, g)
`ministered at a daily dose of 0.5 to 500 mg per kg of
`—3.3
`10/10
`body weight.
`-1.5
`10/10
`5. The method of claim2 wherein rapamycin is ad-
`1.2
`10/10
`ministered at a daily dose of 10 to 250 mg per kg of body
`—0.7
`10/10
`60 Weight.
`0.1
`10/10
`6. The method of reducing tumorsize in a colon
`Ol
`10/10
`tumor bearing mammal, comprising administering to
`said mammal an anti-colon tumor effective amount of
`rapamycin.
`7. The method of prolonging the survival time of a
`colon tumor bearing mammal, which comprises admin-
`istering to said mammal an anti-colon tumoreffective
`amount of rapamycin.*
`*
`*
`
`Effect of Rapamycin on Survival Time of CDF, Mice
`Implanted with Colon 26 Tumor
`Ave. Wt. Difference
`MST
`of Animals
`days
`(T-C, 8)
`~2.4
`—18
`—14
`—0.8
`—0.3
`03
`
`T
`
`Cc
`
`T/C % 20
`MST
`
`Dose/Inj.
`mg/kg
`400
`200
`100
`50
`25
`12.5
`Treatment:
`Single intraperitoneal injection on days1, 5 and 9 in a vehicle ofsaline with Tween-
`80.
`Evaluation:
`T/C % = Median Survival Time (MST) in days of treated animals (T) control 30
`animals (C) X 100. A T/C % of 125 or greater is considered as a significant
`prolongation of host survival. Evaluation done on day 60.
`
`Survivors
`on Day 5
`10/10
`10/10
`10/10
`10/10
`10/10
`10/10
`
`59
`
`Dose/Inj.
`T/C % 55
`mg/kg
`MST
`400
`109
`200
`110
`100
`139
`50
`125
`25
`139
`12.5
`144
`Treatment:
`Single intraperitoneal injection on each of days 1 through 9 in a vehicle ofsaline
`with Tween-80.
`Evaluation:
`T/C % = Median Survival Time (MST) in days of treated animals (T) control
`animals (C) X 100. A T/C % of 125 or greater is considered as a significant
`prolongation of host survival. Evaluation done on day 60.
`
`T
`22.0
`22.3
`28.0
`25.3
`28.0
`29.0
`
`Cc
`20.1
`20.1
`20.1
`20.1
`20.1
`20.1
`
`65
`
`West-Ward Pharm.
`Exhibit 1042
`Page 003
`
`West-Ward Pharm.
`Exhibit 1042
`Page 003
`
`

`

`
`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`CERTIFICATE OF CORRECTION
`4 395 17)
`=
`PATENT NO.
`* December 5, 1989
`DATED
`INVENTOR(S) :
`Surendra N. Sehgal et al
`It is certified that error appears in the above-identified patent and that said Letters Patent is hereby
`corrected as shown below:
`
`On the title page;
`
`Item [19] Sehgal et al.
`
`[75] Inventors: Surendra N. Sehgal, Dollard Des Orneaux;
`Claude Vezina, Oka, both of Canada
`
`[73] Assignee: Ayerst, McKenna & Harrison, Inc. Ville
`St. Laurent, Quebec, Canada
`
`Signed and Sealed this
`
`Twenty-sixth Day of January, 1993
`
`Attest:
`
`Attesting Officer
`
`Acting Commissioner of Patents and Trademarks
`
`STEPHEN G. KUNIN
`
`West-Ward Pharm.
`Exhibit 1042
`Page 004
`
`West-Ward Pharm.
`Exhibit 1042
`Page 004
`
`