United States Patent
`Surendra et al.
`
`[19]
`
`[11]
`
`[45]
`
`Patent Number:
`
`Date of Patent:
`
`4,885,171
`
`Dec. 5, 1989
`
`[54]
`
`[75]
`
`[73]
`
`[21]
`
`[22]
`
`[63]
`
`USE OF RAPAMYCIN IN TREATMENT OF
`CERTAIN TUMORS
`
`Assignee:
`
`Inventors: Sehgal N. Surendra, Dollard Des
`Ormeaux; Claude Vezina, Oka, both
`of Canada
`American Home Products
`Corporation, New York, N.Y.
`592,193
`Mar. 22, 1984
`
`Appl. No.:
`Filed:
`
`Related U.S. Application Data
`Continuation of Ser. No. 126,276, Mar. 3, I980, aban-
`doned, which is a continuation of Ser. No. 957,626,
`Nov. 3, 1978, abandoned.
`
`Int. Cl.‘ .............................................. A61K 35/74
`
`.................................................. .. 424/122
`[52] U.S.Cl.
`[58] Field of Search ......................................... 424/122
`References Cited
`PUBLICATIONS
`
`[56]
`
`Endicott, J. of the National Cancer Institute, The Che-
`motherapy Program, vol. 19, No. 2 (20th Anniversary)
`Aug. 1957, pp. 275-293.
`
`Primary Examiner-—-Jeroine D. Goldberg
`Attorney, Agent, or Firm—Wa.lter Patton
`
`[57]
`
`ABSTRACI‘
`
`Methods for using rapamycin in the treatment of certain
`cancers or tumors are disclosed.
`
`7 Claims, No Drawings
`
`Par Pharm., Inc.
`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 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, 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 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 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
`
`10
`
`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 eta1., 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 20
`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
`[(experimenta1 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 30
`activity of rapamycin appears to be related to inhibition
`of the lymphatic system.
`SUMMARY OF THE INVENTION
`
`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, eg. 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
`
`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
`CD8F1 mammary tumors in male CD8Fl rats by ad-
`ministering rapamycin; Table 5 shows the prolongation
`of survival time of female BDF1 mice implanted with
`B 16 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
`Efiect of Rapamycin on Survival Time CDF1 Mice
`Implanted with Lymphatic Leukemia P-388!ascetic!.
`Ave. WL Difference
`MST
`Survivors
`of Animals
`days
`(T-C. 8)
`on Day 5
`6/6
`-1.9
`6/6
`-2.4
`6/6
`- 1.6
`6/6
`- 1.9
`6/6
`-1.6
`6/6
`-0.6
`
`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
`
`Dose/Inj
`mg/kg
`400
`200
`100
`50
`25
`12.5
`Treatment:
`Nine inlraperitoneal injections staning 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-ethanediy1)].
`Evalualion:
`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 prolon-
`gation of host survival. Evaluation done on day 30.
`
`T/C %
`MST
`138
`128
`134
`140
`136
`136
`
`TABLE 2
`
`Effect of Rapamycin on Colon 38 Tumor Weight in Mice
`Ave. Net Wt.
`Difference
`MTW
`of Animals
`mg
`(T-C. s)
`— 3.4
`-2.0
`-0.8
`-0.8
`-0.4
`
`T/C %
`MTW
`
`Survivors
`Day 5
`10/10
`10/10
`10/10
`9/10
`10/10
`
`T
`188
`209
`272
`320
`368
`
`C
`810
`810
`810
`810
`810
`
`Dose/Inj
`ms/ks
`4-00
`200
`100
`50
`
`Par Pharm., Inc.
`Exhibit 1042
`Page 002
`
`

`
`3
`TABLE 2-continued
`
`4,885,171
`
`Effect of Rapamycin on Colon 38 Tumor Weight in Mice
`Ave. Net Wt.
`Difference
`MTW
`of Animals
`mg
`(T-C, 5)
`0.4
`
`'1‘
`368
`
`c
`810
`
`T/C %
`MTW
`45
`
`Dose/lnj
`ms/ks
`125
`Treatment:
`Single intraperitoneal injection on days 2. 9 and 16 in a vehicle of saline with
`Tween-80.
`Evaluation:
`T/C % = Median tumor weight (MTW) estimated from tumor diameter of treated
`animals (T)/control animals (C) X 100. A T/C % of 42 or less is considered as a
`significant inhibitor of tumor growth. Evaluation done on day 20.
`
`Survivors
`Day 5
`10/10
`
`TABLE 3.
`Effect of Rapamycin on Survival Time of CDF1 Mice
`Implanted with Colon 26 Tumor
`Ave. Wt. Difference
`MST
`
`of Animals
`(T-C, g)
`-2.4
`-1.8
`-1.4
`-0.8
`-0.3
`0.3
`
`Survivors
`on Day 5
`10/10
`10/10
`10/10
`10/10
`10/10
`10/10
`
`days
`C
`T
`26.3
`19.1
`25.8
`19.1
`29.0
`19.1
`30.6
`19.1
`30.3
`19.1
`30.4
`19.1
`
`T/C % 20
`MST
`137
`135
`151
`160
`158
`159
`
`25
`
`Dose/lnj.
`ms/ks
`400
`200
`100
`50
`25
`12.5
`Treatment:
`Single intrxperitoneal injection on days 1, 5 and 9 in a vehicle of saline with Tween-
`30.
`Evaluation:
`T/C % = Median Survival Time (MST) in days of treated animals (1') control 30
`animals (C) X 100. A T/C % of 175 or greater is considered as a significant
`prolongltion of host survival. Evaluation done on day 60.
`
`TABLE 4
`
`4
`TABLE 6
`Effect of Rapamycin on Ependymoblastorna in Swiss Mice
`Ave. Wt. Difference
`MST
`Survivors
`Dose/'lnj.
`of Animals
`days
`ms/ks
`(T-C. 3)
`on day 5
`200
`—-3.3
`10/10
`100
`-2.2
`l0/10
`S0
`-1.3
`9/10
`25
`-2.0
`10/10
`12.5
`-1.0
`10/10
`Treatment:
`Single imnperitoneal injection on each of days 1 through 9 in a vehicle of saline
`with Tween-80.
`Evaluation:
`T/C % = Median Survival Time (MST) in days of treated animals ('1') control
`animals (C) X 100. A T/C % of 125 or greater is considered as a significant
`prolongntion of host survival. Evaluation done on day 60.
`
`T/C %
`
`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-
`lites, for example, cytarabine, fluorouracil, floxuridine,
`mercaptopurine, methotrexate and thioguanine; miscel-
`laneous anticancer agents, for example, dacarbazine,
`hydroxyurea, mitotane, procarbazine hydrochloride,
`quinacrine hydrochloride, vinblastine sulfate and viri-
`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 the like; proges-
`tagens, for example, megestrol, hydroxyprogesterone
`caproate and the like; radioactive isotopes; and antibiot-
`ics, for example, bleomycin sulfate, doxorubicin hydro-
`chloride and the like. Suitable methods of administra-
`tion, compositions and dosages of the antineoplastic
`agents are described in medical textbooks; for instance,
`“PHYSICIANS‘ DESK REFERENCE”, 32nd ed.,
`Medical Economics Co., Oradell, NJ. U.S.A., 1978 and
`“AMA DRUG EVALUATIONS”, 3 ed. PSG Publish-
`ing Company,
`Inc., Littleton, Mass., U.S.A., pp
`1106-1151, 1977. When used in combination, rapamycin
`is administered as described previously; however, a
`lower dose can be used for efficacious results.
`We claim:
`1. A method of treating transplanted tumors in a
`transplanted tumor bearing mammal, wherein said
`tumor is selected from lymphatic leukemia, colon, mam-
`mary, melanocarcinoma and ependymoblastoma tumors
`which comprises administering to said mammal an anti-
`tumor effective amount of rapamycin.
`2. The method of claim 1 wherein rapamycin 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-
`ministered at a daily dose of 0.5 to 500 mg per kg of
`body weight.
`5. The method of claim’2 wherein rapamycin is ad-
`ministered at a daily dose of 10 to 250 mg per kg of body
`weight.
`6. The method of reducing tumor size in a colon
`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 tumor effective
`amount of rapamycin.3
`0
`i
`O
`t
`
`Effect of Rapamycin on cnsr, Mgmggg Tumors in CDBFI Rats
`Ave. Net Wt.
`Difference
`MTW
`of Animals
`mg
`(TC. 9)
`-6.6
`-6.5
`-4.8
`-4.1
`-2.4
`-0.8
`
`T
`0
`323
`448
`755
`825
`928
`
`C
`3200
`3200
`3200
`3200
`3200
`3200
`
`T/C %
`MTW
`
`10
`14
`23
`25
`29
`
`Dose/Inj
`mg/kg
`400
`200
`100
`50
`25
`12.5
`Treatment:
`Single intraperitoneal injection on days 1, 8, 15, 22 and 29 in vehicle of saline with
`Tween-80.
`Evaluation: V
`T/C % = Medien tumor weight (MTW) estimated from tumor diameter of treated
`animnls (T)/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.
`
`Survivors
`Day 5
`4/10
`10/10
`10/10
`10/ 10
`10/10
`10/10
`
`TABLE 5
`Effect of Rapamycin on B 16 melanocarcinoma in BDF} Mice
`Ave. Wt. Difference
`MST
`Dose/Inj.
`T/C %
`Survivors
`of Animals
`days
`mg/kg
`MST
`C
`T
`on Day 5
`(T-C, g)
`400
`109
`20.1
`22.0
`10/10
`-3.3
`200
`110
`20.1
`22.3
`10/10
`-1.5
`1m
`139
`20.1
`28.0
`10/10
`-1.2
`125
`20.1
`25.3
`10/10
`-0.7
`50
`139
`20.1
`28.0
`10/10
`0.1
`25
`
`
`
`
`
`0.1 10/10 29.0 20.112.5 144
`Treatment:
`Single intraperitoneal injection on each of days 1 through 9 in a vehicle of saline
`with Tween-30.
`Evaluation:
`'1'/C % : Median Survival Time (MST) in days of treated animals ('1') 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.
`
`35
`
`4-0
`
`45
`
`50
`
`55
`
`60
`
`65
`
`Par Pharm., Inc.
`Exhibit 1042
`Page 003
`
`

`
`UNITED STATES PATENT AND TRADEMARK OFFICE
`CERTIFICATE OF CORRECTION
`
`PATENT N0.
`
`4 .885.171
`
`DATED
`
`December 5 , 1989
`Surendra N. Sehgal at al
`INVENTDMS) 3
`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
`
`Anesfing 017;;-gr
`
`Acting Commixgioner of Parent: and Trademarks
`
`STEPHEN G. KUNIN
`
`Par Pharm., Inc.
`Exhibit 1042
`Page 004