Europaisches Patentamt
`
`European Patent Office
`
`Office europeen des brevets
`
`I IIIIIII IIIIII Ill lllll lllll lllll lllll lllll lllll lllll lllll 111111111111111111
`0 525 960 A1
`0 Publication number:
`
`@
`
`EUROPEAN PATENT APPLICATION
`
`@ Application number: 92305287.2
`
`@ Int. Cl.5: A61 K 35/74
`
`@ Date of filing: 09.06.92
`
`@ Priority: 18.06.91 US 717773
`
`@ Date of publication of application:
`03.02.93 Bulletin 93/05
`
`@ Designated Contracting States:
`AT BE CH DE DK ES FR GB GR IT LI LU NL PT
`SE
`
`E) Applicant: AMERICAN HOME PRODUCTS
`CORPORATION
`685 Third Avenue
`New York New York 10017-4085(US)
`
`@ Inventor: Miller, Glenn Alvin
`
`9780 NW 51st La.
`Miami, Dade, Florida(US)
`Inventor: Rabin, Mark Barry
`8260 SW 146th Street
`Miami, Dade, Florida(US)
`Inventor: Harrington, William Joseph, Jnr.
`9301 SW 92nd Avenue
`Miami, Dade, Florida(US)
`
`@ Representative: Wileman, David Francis Dr. et
`al
`c/o Wyeth Laboratories Huntercombe Lane
`South
`Taplow Maidenhead Berkshire SL6 OPH(GB)
`
`@ Use of rapamycin for the treatment of adult T-cell leukemia/lymphoma.
`
`© This invention provides a method of treating adult T-cell leukemia/lymphoma (ATL) in a mammal in need
`thereof which comprises administering an antiproliferative amount of rapamycin, and is particularly useful in
`arresting the progression of or inducing remission of ATL.
`
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`EP O 525 960 A1
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`This invention relates to a method of treating adult T-cell leukemia/lymphoma, in particular to a new
`therapeutic use of rapamycin.
`Adult T-cell Leukemia/Lymphoma (ATL) is an aggressive hematologic disease, characterized by a rapid,
`invariably fatal course. Clinical features of ATL include hypercalcemia, bone marrow infiltration, generalized
`lymphadenopathy, leukemia, skin involvement, hepatosplenomegaly and lytic bone lesions.
`The human T-lymphotropic virus 1 (HTL V-1) is the etiologic agent of ATL. Evidence linking HTL V-1 to
`ATL includes epidemiologic clustering of ATL in geographic regions endemic for HTLV-1 infection [Hinuma,
`Y., Proc. Natl. Acad. Sci. 78: 6476 (1981 ); Watanabe, T., Virology 133: 238 (1984); Catovsky, D. Lancet 1:
`639 (1982); Blattner, W.A., Int. J. Ca. 30: 257 (1982)], the high frequency of specific antibodies in sera from
`10 ATL patients, [Hinuma, Y., Proc. Natl. Acad. Sci. 78: 6476 (1981 )], detection of monoclonally integrated
`proviral sequences in leukemic cells of patients with this disease [Yoshida M., Proc. Natl. Acad. Sci. 81:
`2534 (1984)], and isolation of infectious virus from leukemia cell cultures [Poiesz, B.J., Proc. Natl. Acad. Sci.
`77: 7415 (1980); Yoshida, M., Proc. Natl. Acad. Sci. 79: 2031 (1982)].
`HTL V-1 transformed T-cells are characterized by uncontrolled proliferation in the absence of antigenic
`stimulation. IL-2 independent growth is a hallmark of these cells in culture. The cell surface phenotype of
`these cells is dominated by a significant increase in IL-2 receptors. Recent evidence demonstrated that
`these receptors are high affinity IL-2R. The majority of T-cells isolated from infected individuals tested did
`not, however, respond to exogenously added IL-2. [Kodaka, T., Jpn. J. Ca. Res. 81: 902 (1990)].
`HTLV-1 has been reported to be transmitted sexually, from mother-childvla breast feeding, among
`intravenous drug abusers (presumably by sharing of contaminated needles), and via blood transfusion
`[Hino, S., Gann 76: 474 (1985); Murphy, E.L., Ann. Int. Med. 111: 555 (1989); Sato, H., Int. J. Ca. 37: 395
`(1986); Robert-Guroff, M., JAM.A., 255: 3133 (1986)]. Infection with the virus has an associated lifetime risk
`for developing ATL estimated to be approximately 4% [Murphy, E.L., Ann. Int. Med. 111: 555 (1989)].
`The prevalence of HTLV-1 infection in residents of the United States has recently become a public
`health concern. An epidemiologic survey undertaken in major cities of the United States detected average
`seroprevalence rates of 0.025 percent among healthy blood donors [Williams, A.E., Science, 240: 643
`(1988)]. More recent epidemiologic data indicate a seropositivity rate of 4.4 percent among healthy blacks
`in Brooklyn, New York suggesting seroprevalence may vary significantly depending on the population
`sampled. [Dosik, H., Proc. Am. Soc. Clin. Oncol. 9: 2 (1990)]. Khabbaz [JAM.A. 263: 60 (1990)] reported
`30 average seroprevalence rates of 6.7% among prostitutes nationwide. These data ranged from a low of 0%
`in Nevada to a high of 25.4% in Newark, NJ indicating variable rates even among high risk groups.
`Internationally, HTLV-1 is present in endemic proportions in several areas of the world. The southwest(cid:173)
`ern islands of Japan have long been identified as an endemic region for HTLV-1 infection. [Jpn. J. Clin.
`Oncol. 15: 517 (1985)]. For example, seroprevalence rates as high as 8% have been detected in healthy
`volunteer blood donors in the southern providence of Kyushu. [Maeda, Y., Int. J. Ca. 33: 717 (1984)]. A
`seroprevalence rate of 9.2% among otherwise healthy female food service workers over the age of 30 was
`reported in Jamaica. [Murphy, E.L., Ann. Int. Med. 111: 555 (1989)]. The seroprevalence rate rose to 14.1 %
`in age matched females seen at a Jamaican sexually transmitted disease clinic. Risk factors for infection
`with the virus follow the expected pattern of a sexually transmitted disease.
`Although serological and molecular genetic tests are available to screen for HTLV-1 infection, there is
`currently no means of early diagnosis of ATL. Presently, there is no effective therapeutic treatment for late
`stage disease, and as such mean survival after diagnosis is less than 6 months. Unsuccessful treatments
`were reported using radiation; methotrexate; leukovorin; interferon SQ; high dose steroids; and combinations
`containing cyclophosphamide, adriamycin, oncovin, prednisone, and bleomycin therapies. [Harrington, J.
`45 Aids 4: 284 (1991 )]. Efficacious treatment of ATL was reported with SUN4800 (recombinant interferon
`gamma). [Ishihara, K., J. Invest. Dermatol. 93: 556 (1989)]. Responses of 90% were reported in 3 of 10 ATL
`patients treated with anti-Tac, a mouse antibody against the human p55 IL-2 receptor peptide (Tac; CD25).
`[Junghans, R., 37: 861 A (1989)].
`Rapamycin, a macrocyclic triene antibiotic produced by Streptomyces hygroscopicus [U.S. Patent
`50 3,929,992] has been shown to prevent the formation of humoral (lgE-like) antibodies in response to an
`albumin allergic challenge [Martel, R., Can. J. Physiol. Pharm. 55: 48 (1977)], inhibit murine T-cell activation
`[Strauch, M., FASEB 3: 3411 (1989)], and prolong survival time of organ grafts in histoincompatible rodents
`[Morris, R., Med. Sci. Res. 17: 877 (1989)].
`This invention provides a method of treating adult T-cell leukemia/lymphoma (ATL) in a mammal in
`55 need thereof which comprises administering an antiproliferative amount of rapamycin orally, parenterally,
`In particular, rapamycin is useful in
`intranasally, intrabronchially, topically, transdermally, or rectally.
`arresting the progression of and inducing remission of ATL by virtue of its ability to inhibit the proliferation
`of HTLV-1 transformed T-cells.
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`This invention also provides use of rapamycin in the preparation of a medicament for treating adult T(cid:173)
`cell leukemia/lymphoma in a mammal.
`The effect of rapamycin on ATL was established in an in vitro standard pharmacological test procedure
`using ten human HTLV-1 transformed T-cell lines cultured from ATL patients. These cell lines are presently
`the best models for ATL in humans. Cyclosporin A (CsA) was also evaluated for the purpose of comparison.
`The procedure used and results obtained are described below.
`ATL cell lines were established according to known literature procedures. ATL5S, ATL3I, and ATL 1 K
`were established according to the method of Hoshino [Proc. Natl. Acad. Sci, 80: 6061 (1983)]. The cell lines
`31008, 31009, 31016 and AT(EGG) were established from peripheral blood of HTLV-1 infected individuals
`via co-culture with cord blood lymphocytes. The cell line MALN was isolated from lymphoid tumor tissue of
`an HTLV-1 infected ATL patient seen at Jackson Memorial/University of Miami Medical Center. [Harrington,
`W.J., Jr., J. Acq. Imm. Def. Dis., 4: 284 (1991 )]. The primary cultures BL and GP were initiated from
`peripheral blood of ATL patients. The MALN, BL, and GP cell lines were initiated without cultivation.
`Cells were adjusted to a concentration of 1x106 cells/ml in RPMI 1640 with 15% fetal bovine serum
`and were supplemented with 50 U/ml penicillin, 50 I.Lg/ml streptomycin, and 2 mM glutamine. Rapamycin
`and CsA were each diluted to a stock concentration of 1 mM in absolute ethanol with subsequent dilutions
`being made in culture medium. Drug concentrations evaluated ranged from 11.LM to 100 fM. Cultures were
`incubated at 37 ° C, 5% CO2 for 24 hours followed by a 4 hr. pulse with 0.5 1.LCi of tritiated thymidine.
`Thymidine incorporation was measured by liquid scintillation counting, and the results expressed in counts
`20 per minute (cpm). The results obtained from the mean of four evaluations at each concentration were
`compared with control values to determine percent inhibition. The following formula was used to calculate
`the percent inhibition:
`
`10
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`15
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`25
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`30
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`35
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`40
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`45
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`Percent Inhibition
`
`= cpm[no drug] - cpm[drug] x 100
`cpm[no drug]
`
`The percent inhibition obtained with rapamycin is shown in the following table.
`
`PERCENT INHIBITION OF ATL CELL PROLIFERATION
`
`Rapamycin Concentration
`
`Cell Line
`
`1 1.LM
`100 nM
`10 nM
`1 nM
`100 pM
`10 pM
`1 pM
`100 fM
`
`ATL3I ATL5S ATL1K ATEGG 31008 31009 31016
`
`MALN GP
`
`BL
`
`28
`17
`15
`18
`11
`6
`11
`7
`
`59
`58
`49
`48
`47
`38
`14
`36
`
`19
`32
`18
`26
`23
`12
`15
`15
`
`58
`60
`45
`53
`49
`44
`48
`44
`
`69
`68
`58
`54
`45
`31
`30
`29
`
`71
`70
`67
`68
`61
`47
`59
`60
`
`59
`56
`49
`48
`49
`47
`50
`44
`
`55
`54
`41
`43
`37
`26
`36
`36
`
`54
`46
`46
`42
`39
`14
`20
`23
`
`80
`79
`76
`73
`65
`55
`56
`51
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`Rapamycin inhibited proliferation of 8 of the 10 ATL cell lines at a concentration of 100 nM or greater,
`with a 50% reduction in proliferation in 4 of the cell lines at a concentration of 1 nM. Both of the cell lines in
`which proliferation was not inhibited by rapamycin had been in culture for more than 5 years without
`recloning; their nonresponsiveness to rapamycin may be attributed to their further transformation into
`insensitive clones during this time period. In comparison, CsA did not inhibit proliferation in 7 of the 10 cell
`lines in which it was evaluated. In the remaining 3 cell lines (ATL3I, ATL5S, and 31009) proliferation was
`inhibited by 16%, 23%, and 22%, respectively.
`The results obtained in this standard pharmacological test procedure demonstrate that rapamycin
`inhibits the proliferation of HTLV-1 transformed T-cell lines, which are presently the best models for ATL in
`humans, and as such, rapamycin is useful in the treatment of ATL.
`When rapamycin is employed in the treatment of ATL, it can be formulated neat or with a pharmaceuti(cid:173)
`cal carrier to a mammal in need thereof preferably in unit dosage form; conveniently a single unit dose for
`daily administration. The pharmaceutical carrier may be solid or liquid.
`A solid carrier can include one or more substances which may also act as flavoring agents, lubricants,
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`solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it
`can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in admixture
`with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier having the
`necessary compression properties in suitable proportions and compacted in the shape and size desired.
`5 Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose,
`dextrin, starch, gelatin, cellulose, methyl cellulose, sodium carboxymethyl cellulose, polyvinylpyrrolidine, low
`melting waxes and ion exchange resins.
`Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized
`compositions. The active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid
`carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats. The
`liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers,
`preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regula(cid:173)
`tors, stabilizers or osmo-regulators. Suitable examples of liquid carriers for oral and parenteral administra(cid:173)
`tion include water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium
`carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g.
`glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil). For parenteral
`administration, the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate. Sterile
`liquid carriers are useful in sterile liquid form compositions for parenteral administration. The liquid carrier
`for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable
`20 propellent.
`Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by, for
`example, intramuscular, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered
`intravenously. The compound can also be administered orally either in liquid or solid composition form.
`Rapamycin may be administered rectally in the form of a conventional suppository. For administration
`25 by intranasal or intrabronchial inhalation or insufflation, rapamycin may be formulated into an aqueous or
`partially aqueous solution, which can then be utilized in the form of an aerosol. Rapamycin may also be
`administered transdermally through the use of a transdermal patch containing the active compound and a
`carrier that is inert to the active compound, is non-toxic to the skin, and allows delivery of the agent for
`systemic absorption into the blood stream via the skin. The carrier may take any number of forms such as
`creams and ointments, pastes, gels, and occlusive devices. The creams and ointments may be viscous
`liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive
`powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may also be
`suitable. A variety of occlusive devices may be used to release the active ingredient into the blood stream
`such as a semipermiable membrane covering a reservoir containing the active ingredient with or without a
`carrier, or a matrix containing the active ingredient. Other occlusive devices are known in the literature.
`Rapamycin may be administered topically as a solution, cream, or lotion by formulation with phar(cid:173)
`maceutically acceptable vehicles containing 0.1 - 5 percent, preferably 2%, of active compound.
`The dosage requirements vary with the particular compositions employed, the route of administration,
`the severity of the symptoms presented and the particular subject being treated. Based on the results
`40 obtained in the standard pharmacological test procedure, projected parenteral daily dosages of active
`compound would be 0.01 I.Lg/kg - 10 mg/kg and preferably between 0.1 I.Lg/kg - 2 mg/kg. When rapamycin
`is given parenterally, it will preferably be administered intravenously. Projected oral daily dosages of active
`compound would be 1 I.Lg/kg - 15 mg/kg and preferably between 0.01 mg/kg - 10 mg/kg. Treatment will
`generally be initiated with small dosages less than the optimum dose of the compound. Thereafter the
`45 dosage is increased until the optimum effect under the circumstances is reached; precise dosages for oral,
`parenteral, nasal, or intrabronchial administration will be determined by the administering physician based
`on experience with the individual subject treated. In general, rapamycin is most desirably administered at a
`concentration that will generally afford effective results without causing any harmful or deleterious side
`effects, and can be administered either as a single unit dose, or if desired, the dosage may be divided into
`convenient subunits administered at suitable times throughout the day.
`For the treatment of ATL, it is also contemplated that rapamycin may be administered in conjunction
`with chemotherapeutic agents such as methotrexate, leukovorin, cyclophosphamide, adriamycin, oncovin,
`prednisone, bleomycin, and the like; high dose steroids; radiation therapy; cytokine therapy, such as
`interferon; monoclonal antibody therapy; and other therapies useful in the treatment of neoplastic diseases.
`
`50
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`55
`
`Claims
`
`1. Use of rapamycin in the preparation of a medicament for treating adult T-cell leukemia/lymphoma in a
`
`4
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`

`EP O 525 960 A1
`
`mammal.
`
`2. Use of rapamycin according to Claim 1 in which the medicament is adapted for administration orally,
`parenterally, intranasally, intrabronchially, topically, transdermally or rectally.
`
`5
`
`3. Use of rapamycin according to Claim 2 in which the medicament is adapted for intravenous administra(cid:173)
`tion.
`
`4. Use of rapamycin according to Claim 3 in which the medicament is in unit dosage form to provide a
`daily dosage from 0.01 I.Lg/kg to 10 mg/kg based on the weight of the mammal to treated.
`
`10
`
`5. Use of rapamycin according to Claim 3 in which the medicament is in unit dosage form to provide a
`daily dosage from 0.1 I.Lg/kg to 2 mg/kg based on the weight of the mammal to treated.
`
`15
`
`6. Use of rapamycin according to Claim 1 in which the medicament is adapted for oral administration.
`
`7. Use of rapamycin according to Claim 6 in which the medicament is in unit dosage form to provide a
`daily dosage from 1 I.Lg/kg to 15 mg/kg based on the weight of the mammal to treated.
`
`20 8. Use of rapamycin according to Claim 6 in which the medicament is in unit dosage form to provide a
`daily dosage from 0.01 mg/kg to 10 mg/kg based on the weight of the mammal to treated.
`
`25
`
`30
`
`35
`
`40
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`5
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`

`

`European Patent
`Office
`
`EUROPEAN SEARCH REPORT
`
`Category
`
`DOCUMENTS CONSIDERED TO BE RELEVANT
`Citation of document with indication, where appropriate,
`of relevant passages
`P, X THE JOURNAL OF IMMUNOLOGY, vol . 148,
`no. 10, 15th May 1992, pages 3256-3263,
`The American Association of
`Immunologists, US; P. HOLLSBERG et al.:
`11 Characterization of HTLV-I in vivo
`infected T cell clones.
`IL-2-independent growth of
`nontransformed T cells 11
`* Whole document*
`
`Relevant
`to claim
`1
`
`Application Number
`
`EP 92 30 5287
`
`CLASSIFICATION OF TIIE
`APPLICATION (Int. CI.S)
`
`A 61 K 35/74
`
`1-8
`
`1-8
`
`A
`
`A
`
`THE JOURNAL OF IMMUNOLOGY, vol. 144,
`no. 4, 15th February 1990, pages
`1418-1424, The American Association of
`Immunologists, US; F.J. DUMONT et al.:
`11The immunosuppressive macrolides
`FK-506 and rapamycin act as reciprocal
`antagonists in murine T cells 11
`* Abstract; page 1423, lines 42-53 *
`
`TRANSPLANTATION PROCEEDINGS, vol. 23,
`no. 2 (suppl. 2), 1991, pages 1-5,
`PROCEEDINGS OF A SATELLITE SYMPOSIUM ON
`IMMUNOSUPPRESSIVE DRUGS: TARGETS AND
`MECHANISMS OF ACTION, San Francisco,
`CA, 19th August 1990; N.H. SIGAL et
`al.: 11 Inhibition of human T-cell
`activation by FK 506, rapamycin, and
`cyclosporine A"
`* Whole document*
`
`-/-
`
`TECHNICAL FIEIDS
`SEARCHED (Int. CI.S)
`
`A 61 K
`
`The present search report bas been drawn up for all claims
`
`Place of search
`
`Dale of oomplellon of Ille ,earth
`
`-
`
`I
`I
`MAIR J.
`28-09-1992
`THE HAGUE
`-
`(i----________ _.____ __ -----1
`~
`8
`~
`
`CATEGORY OF OTED DOCUMENTS
`
`X : particularly relevant if taken alone
`Y : particularly relevant if combined with another
`document of the same category
`A : technological background
`0 : non-written disclosure
`P : intermediate document
`
`T : theory or principle underlying the invention
`E : earlier patent document, but published on, or
`after the filing date
`D : document cited in the application
`L : document cited for other reasons
`
`& : member of the same patent family, corresponding
`document
`
`······----············ ...... ---··· .. ······ .. ----
`~ .. 0 ...
`""~-----------------------------------~
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`

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`Page
`
`2
`
`Application Number
`
`EP 92 30 5287
`
`CLASSIFICATION OF THE
`APPLICATION (Int. Cl.5)
`
`European Patent
`Office
`
`EUROPEAN SEARCH REPORT
`
`Category
`
`P,X
`
`DOCUMENTS CONSIDERED TO BE RELEVANT
`Citation of document with indication, where appropriate,
`of relevant passages
`JOURNAL OF NEUROIMMUNOLOGY, vol. 0,
`suppl. 1, 1991, page 201, 3RD
`INTERNATIONAL CONFERENCE ON
`NEUROIMMUNOLOGY, Jerusalem, 27th
`October - 1st November 1991; P.
`HOLLSBERG et a 1. : 11 HTLV-l induced
`spontaneous T cell clonal proliferation
`is rapamycin sensitive 11
`* Whole document*
`
`Relevant
`to claim
`1
`
`A
`
`A
`
`TRENDS IN PHARMACOLOGICAL SCIENCES,
`vol. 12, no. 6, June 1991, pages
`218-223, Elsevier Science Publishers
`Ltd, GB; J.Y. CHANG et al.: 11 FK-S06 and
`rapamycin: novel pharmacological probes
`of the immune response 11
`* Whole document*
`(AYERST, McKENNA &
`
`877 700
`BE-A-
`HARRISON, INC.)
`* Whole document*
`
`1-8
`
`1-8
`
`TECHNICAL FIELDS
`SEARCHED (Int. Cl.5)
`
`The present search report bas been drawn up for all claims
`
`I
`f ~_T_H_E _H_AG_U_E ____ _,__ ___ 28_-_09_-_1_99_2 ___ _.___M_AI_R_J_. _____ --1
`I
`~
`
`Place of search
`
`Dale of completion of tbe sean:lt
`
`Examiner
`
`CATEGORY OF OTED DOCUMENTS
`
`T : theory or principle underlying the invention
`E : earlier patent document, but published on, or
`after the filing date
`X : particularly relevant if taken alone
`~
`D : document cited in the application
`Y : particularly relevant if combined with another
`-
`L : document cited for other reasons
`document of the same category
`~
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