United States Patent [19]
`Surendra et al.
`
`[54] USE OF RAPAMYCIN IN TREATMENT OF
`CERTAIN TUMCRS
`[75] Inventors: Sehgal N. Surendra, Dollard Des
`Ormeaux; Claude Vezina, Oka, both
`of Canada
`American Home Products
`Corporation, New York, N.Y.
`[21] Appl. No.: 592,193
`[22] Filed:
`Mar. 22, 1984
`
`[73] Assignee:
`
`[63)
`
`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.
`[51] Int. Cl* .............................................. A61K 35/74
`
`[11] Patent Number:
`[45] Date of Patent:
`
`4,885,171
`Dec. 5, 1989
`
`[52] U.S. Cl. .................................................... 424/122
`[58] Field of Search ......................................... 424/122
`[56]
`References Cited
`PUBLICATIONS
`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
`[57]
`ABSTRACT
`Methods for using rapamycin in the treatment of certain
`cancers or tumors are disclosed.
`
`7 Claims, No Drawings
`
`Ex. 1062-0001
`
`

`
`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 tumor size 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, rnelanocarcinoma 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 references therein. In
`addition the protocols for the antitumor tests are avail-
`able from the National Cancer Institute, Bethesda, Md.,
`U.S.A.
`
`USE OF RAPANIYCIN 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.
`
`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 et al., J. Antibiot., 28, 721 (1975), S. N. Sehgal et
`al., J. Antibiot., 28, 727 (1975) and S. N. Sehgal et al.,
`U.S. Pat. No. 3,929,992, issued Dec. 30, 1975, filed Apr.
`12, 1974. Rapamycin is extracted from a streptomycete
`(Streptomyces hygrascopicus) isolated from an Easter
`Island soil sample and is 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 adjuvant arthritis (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 appears to be related to inhibition
`of the lymphatic system.
`SUMMARY OF THE INVENTION
`
`5
`
`10
`
`20
`
`30
`
`According to this invention a method is provided for
`treating carcinogenic tumors in a mammal which com-
`prises administering to the mammal an antitumor effec-
`tive amount of rapamycin. More specifically, rapamy-
`cin reduces tumor size in and prolongs the survival time
`of tumor bearing mammals.
`DETAILS OF THE INVENTION
`
`According to the present method, rapamycin is em-
`ployed as the active agent. The isolation 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 tumor bear-
`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. However as 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 tumor size is achieved without
`causing any harmful or deleterious side effects. The
`
`50
`
`60
`
`65
`
`Tables 1 to 6 show the effects of therapy with rapa-
`mycin on various tumor or cancers in rodents.
`More specifically, Table 1 shows the prolongation of
`survival time of female CDF1 mice implanted with
`lymphatic leukemia P338 by administering rapamycin;
`Table 2 shows the reduction in size of colon 38 tumors
`in female BDF1 mice by administering rapamycin;
`Table 3 shows the prolongation of survival time of male
`CDF1 mice implanted with colon 26 tumors by adminis-
`tering rapamycin; Table 4 shows the reduction in size of
`CDSF1 mammary tumors in male CD8Fl rats by ad-
`ministering rapamycin; Table 5 shows the prolongation
`of survival time of female BDF1 mice implanted with
`B16 melonocarcinoma by administering rapamycin; and
`Table 6 shows the prolongation of survival time of male
`Swiss mice implanted with ependymoblastoma by ad-
`ministering rapamycin.
`TABLE 1
`Effect of Rapamycin on Survival Time CDF1 Mice
`Implanted with Lymphatic Leukemia P-388§ascetic).
`Ave. Wt. Difference
`MST
`Survivors
`of Animals
`days
`(T-C. g)
`on Day 5
`6/6
`-1.9
`6/6
`-2.4
`6/6
`- 1.6
`6/6
`- 1.9
`-1.6
`6/6
`-0.6
`6/6
`
`Dose/lnj
`ms/ks
`4-00
`200
`100
`50
`25
`12.5
`Treatment:
`Nine intnperitoneal 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-etha.nediyl)].
`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 2 significant prolon-
`gation of host survival. Evaluation done on day 30.
`
`T
`14.1
`13.1
`13.7
`14.3
`13.9
`13.9
`
`c
`10.2
`10.2
`10.2
`10.2
`10.2
`10.2
`
`T/C %
`MST
`138
`128
`134
`140
`136
`136
`
`TABLE 2
`
`Survivors
`Day 5
`10/10
`10/10
`10/10
`9/10
`10/10
`
`Dose/Inj
`mg/kg
`400
`200
`100
`50
`25
`
`Effect of Rapamycin on Colon 38 Tumor Weight in Mice
`Ave. Net Wt.
`Difference
`MTW
`of Animals
`mg
`(TC. 2)
`-3.4
`-2.0
`-0.8
`-0.8
`-0.4
`
`T/C %
`MTW
`
`T
`188
`209
`272
`320
`368
`
`C
`810
`810
`810
`810
`810
`
`Ex. 1062-0002
`
`

`
`4,885,171
`
`3
`TABLE 2-continued
`Effect of R8 m cm on colon 38 Tumor Wei ht in Mice
`__jL
`Ave Net wt
`Difference
`o
`urvi
`o
`ose n_i
`T/C 7
`'vors
`fAnimals
`D /1 '
`MTW
`C
`T
`Day 5
`(T41 B)
`ms/ks
`45
`810
`368
`10/10
`0.4
`12.5
`Treatment:
`Single intrnperitoneal injection on days 2. 9 and 16 in a vehicle of saline with
`TWBCI1-311
`Evaluation:
`T/C % = Median tumor weight (MTW) estimated from tumor diameter oftreated
`animals (T)/control animals (c) x 100. A '1'/c in of 42 or less is considered as a
`significant inhibitor of tomor growth. Evaluation done on day 20.
`
`s
`
`MTW
`g
`ns
`
`4
`TABLE 6
`Effect of Rapamycin on Ependymoblastoma in Swiss Mice
`.
`Ave. Wt. Difference
`MST
`of Animals
`Survivors gdagj T/C %
`Cm’ 5)
`°" day 5
`T
`MST
`—3.s
`10/10
`44.
`243
`-12
`10/10
`26.
`143
`-1.3
`9/10
`34.0
`187
`_2.0
`10/10
`34.0
`187
`3
`— 1.0
`10/10
`32.
`
`0O
`
`.
`.
`_
`
`Dose/Inj.
`“mg
`200
`100
`50
`25
`12.5
`1-matmcm:
`Single iniraperitoneal injection on each of days 1 through 9 in a vehicle of saline
`with '1'w¢¢n.80.
`Evaluation:
`‘
`_
`_
`_
`_
`T/C % = Median Survival Tune (M51) In days of treated animals (T) control
`331111515 (C) X 100- A T/C % Of 125 01' $|’¢3'¢1' 15 ‘1°"5id°|'=d 35 3 518113513"!
`prolongation of host survival. Evaluation done on day 60.
`
`MST
`137
`135
`151
`15°
`:33
`
`_
`
`3200
`3200
`3200
`3200
`3200
`
`44s
`755
`825
`923
`
`Survivors
`on Day 5
`- gig
`10/10
`10/10
`10/10
`10/10
`
`'
`
`T
`3(3)
`2g_Q
`25.3
`23.0
`29.0
`
`c
`33%
`20_1
`20.1
`20.1
`20.1
`
`50
`
`65
`
`Rapamycin also can be used to produce beneficial
`TABLE 3.
`effects in the treatment of malignant tumors when Com-
`Effect of Rapmycin on Survival Time of CDFI Mice
`.
`.
`.
`.
`2
`bined with a therapeutically effective amount of an
`m Imcd with CD10“ 26 Tumor
`antmeoplastic agent commonly used in cancer therapy.
`Ave. wt Difimncc
`MST
`20 .:‘.11¥,‘,:*1:‘:.g.
`om -r-1--1
`phamide, mechlorethamine hydrochloride, melphalan,
`C
`T
`°“ Day 5
`“"5/kg
`(T‘C‘ 5)
`pipobroman, thiotepa and uracil mustard; antimetabo-
`19.1
`26.3
`I0/10
`400
`-2.4
`lites, for example, cytarabine, fluorouracil, floxuridine,
`19-1
`25-3
`10/10
`200
`-1-8
`mercaptopurine, methotrexate and thioguanine; miscel-
`19-1
`29-0
`10/10
`100
`-1-4
`25 laneous anticancer agents, for example, dacarbazine,
`19-‘
`3°-5
`10/10
`5°
`‘'°-3
`hydroxyurefi, dmitcifltanea pro_c%1ibatz_ine ll11l¥‘dl'0Cl'll‘(i)I‘l@€,
`1:‘:
`33::
`13;:3’
`3 5
`‘$3
`qumacrine y me on e, vm as me 5
`ate an
`vin-
`'
`'
`'
`'
`cristine sulfate; estrogens, for example, chlorotriam'-
`.
`_
`p
`_
`_
`1'.r=-tm.em=
`_
`_
`_
`sen?’ conjugate estrogens (€.g.
`®), dieth-
`Sgngle intrnperitoneal lI'l_|CCtl0n on days 1, 5 and 9 in a vehicle ofsaline with Tween-
`ylstilbestrol and the like; androgens, for example, meth-
`E,,'_mmm.
`T/C % = Median Survival Time (MST) in days of treated animals (1') control 30 yltestosterone, testosterone and the like; adrenal corti-
`animls (<9)
`>< 100. A "If/C % of 12} or greater is considered as a significant
`costeroids, for example, prednisone and the like; proges-
`prolongntion of host survivali Evaluation done on day 60.
`tagens’ for example’ megestrol, hydroxyprogesterone
`.
`caproate and the like; radioactive isotopes; and antibiot-
`ics, for example, bleomycin sul ate, doxorubicin hydro-
`_
`TABLE 4
`chloride and the like. Suitable methods of administra-
`.
` 35 tion, compositions and dosages of the antineoplastic
`AW_=- Net Wt
`agents are described in medical textbooks; for instance,
`D'”=1’_="C=
`“PHYSICIANS’ DESK REFERENCE”, 32nd ed.,
`°f A“‘“““‘
`Medical Economics Co., Oradell, N.J. U.S.A., 1978 and
`(T-C. 2)
`“AMA DRUG EVALUATIONS”, 3 ed. PSG Publish-
`-55
`ing Company,
`Inc., Littleton, Mass., U.S.A., pp
`_5.5
`1106-1151, 1977. When used in combination, rapamycin
`-4.:
`is administered as described previously; however, a
`-4.1
`lower dlose can be used for efficacious results.
`-2.4
`we c aim;
`-03
`1. A method of treating transplanted tumors in a
`transplanted tumor bearing mammal, wherein said
`tumgr
`slelgcted f'fom lymgjhatic ljukel-€33, cQ]on, mam.
`mary, me ano_carcmon_ia_ an 'epen ymo astoma tumors
`Whlch C°mP}'15e-5 admlmstenng to
`mammal an ‘inn’
`11111101’ €ff8Ct1Ve
`811101111! Of 1'apamycin.
`2_ The method of claim 1 wherein rapamycjn is ad-
`ministered orally or parenterally.
`.3.‘ The method of claim 2 wherein rapamycin is ad-
`ministered intraperitoneally as a solution in saline with a
`derivative of (Z)-sorbitan mono-9-octadecenoate poly-
`(oxy-1,2-ethanediyl).
`4. The method of claim 2 wherein rapamycin is ad-
`T/C % 55
`ministered at a daily dose of 0.5 to 500 mg per kg of
`MST
`'— body weight.
`ii:
`5. The method of claim/2 wherein rapamycin is ad-
`139
`ministered at 3. daily dose Of 10 to 250 mg per kg of body
`125
`so Weight-
`139
`6. The method of reducing tumor size in a colon
`144
`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
`mammal’
`comprises admin-
`istering to Said mammal an anti-CO10I1 tumor effective
`amount of rapamyc1n_t
`i
`t
`*
`t
`
`_
`D°’°/1“!
`ms/ks
`4m
`200
`100
`50
`25
`12-5
`Treatment:
`irsignpcritoneal injection on days 1, 8, 15, 22 and 29 in vehicle ofsaline with
`Emu_;im'“
`T/C % = Median tumor weight (MTW) estimated from tumor diameter of treated
`animals ('1')/control animals (C) X 100. A T/C% of 42 or less is considered as a
`significant inhibitor of growth. Evaluation done on day 30.
`
`TABLE 5
`-—j—:—j.——j—-j..j..:..
`£55601 °f_R=_2aII1x°in on 3 16 mclanocarcinoma in BDFi Mice
`Ave. Wt. Difference
`MST
`of Animals
`days
`Cr-c, g)
`_ '
`:3
`_1_2
`-0.7
`0.1
`0.1
`
`Dose/Inj.
`mg/kg
`$3
`100
`50
`zs
`12.5
`1-,,,,..,,¢,,.,
`Single intraperitoneal injection on each of days 1 through 9 in a vehicle of saline
`
`T/C % = Median Survival Time (MST) in days of treated animals ('1') control
`animal: (C) x 100. A T/C % of 125 or greater is considered as a significant
`prolongation of host survival. Evaluation done on day 60.
`
`Ex. 1062-0003
`
`

`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`CERTIFICATE OF CORRECTION
`PATENT NO. : 4,885,171
`DATED
`December 5, 1989
`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 show; 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
`
`DOUGLAS B. COMER
`
`
`
`Ex. 1062-0004
`
`

`
`UNITED states PATENT AND TRADEMARK office
`CERTIFICATE OF CORRECTION
`PATENT NO. : 4.885,171
`DATED
`December 5, 1989
`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. Wille
`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
`
`|
`
`
`
`Ex. 1062-0005