IkAU0
`
`~Office
`
`PCTIiCP 0 2 / 0 i 7
`
`INVESTOR IN PEOPLE
`
`The Patent Office
`Concept House
`Cardiff Road
`Newport
`South Wales
`NP10 8QQ
`
`I, the undersigned, being an officer duly authorised in accordance with Section 74(1) and (4)
`of the Deregulation & Contracting Out Act 1994, to sign and issue certificates on behalf of the
`Comptroller-General, hereby certify that annexed hereto is a true copy of the documents as
`originally filed in connection with the patent application identified therein.
`
`In accordance with the Patents (Companies Re-registration) Rules 1982, if a company named
`in this certificate and any accompanying documents has re-registered under the Companies Act
`1980 with the same name as that with which it was registered immediately before re-
`registration save for the substitution as, or inclusion as, the last part of the name of the words
`"public limited company" or their equivalents in Welsh, references to the name of the company
`in this certificate and any accompanying documents shall be treated as references to the name
`with which it is so re-registered.
`
`In accordance with the rules, the words "public limited company" may be replaced by p.l.c.,
`plc, P.L.C. or PLC.
`
`Re-registration under the Companies Act does not constitute a new legal entity but merely
`subjects
`any to certain additional company law rules.
`
`-
`
`Signed
`
`Dated
`
`8 February 2002
`
`0U
`
`An ExecutiveAgency of the Department of Trade and Industry
`
`Ex. 1085-0001
`
`

`

`EX. 1085—0002
`
`Ex. 1085-0002
`
`

`

`9
`
`E666N5~
`
`DO
`
`"1
`
`----
`
`--
`
`Request for graiff6rba patent
`(See the notes on the back of this form. You can also
`get an explanatory leaflet from the Patent Office to
`help you fill in this form)
`
`1.
`
`2.
`
`3.
`
`Your reference
`
`( h P a e t O f e w l1
`Patent application number
`i p
`(The Patent Office will fill in this pa
`
`4-31671 P3
`
`0145 2
`2l9nt
`
`Full name, address and postcode of the
`or of each applicant
`(underline all surnames)
`
`Patent ADP number (if you know it)
`If the applicant is a corporate body,
`give
`the
`country/state
`of
`its
`incorporation
`
`NOVARTIS AG
`LICHTSTRASSE 35
`4056 BASEL
`SWITZERLAND
`
`SWITZERLAND
`
`4.
`
`Title of invention
`
`Organic Compounds
`
`The Patent Office
`
`Cardiff Road
`Newport
`Gwent NP10 8QQ
`
`pjr' fo
`
`OT20
`
`~1G~ ~WAOQS
`
`5.
`
`Name of your agent (If you have one)
`
`"Address for service" in the United
`Kingdom to which all correspondence
`should be sent
`(including the postcode)
`
`B.A. YORKE & CO.
`CHARTERED PATENT AGENTS
`COOMB HOUSE, 7 ST. J N'S ROAD
`ISLEWORTH
`MIDDLESEX TW7 6N
`
`6.
`
`7.
`
`8.
`
`Patents ADP number (if you know it)
`
`If you are declaring priority from one
`ore more earlier patent applications,
`give
`the country and the date of filing of
`the or of each of these earlier
`applications and (if you know it) the or
`each application number
`
`If this application is divided or
`otherwise derived from an earlier UK
`application, give the number and the
`filing date of the earlier application
`
`1800001
`
`Country
`
`Priority application
`number
`(if you know it)
`
`Date of filing
`(day/month/year
`
`Number of earlier
`application
`
`Date of filing
`(day/month/year)
`
`Is a statement of inventorship and of
`right to grant of a patent required in
`support of this request? (Answer 'Yes' if:
`
`Yes
`
`a) any applicant named in part 3 is not an
`inventor, or
`
`b) there is an inventor who is not named as
`an applicant, or
`
`c) any named applicant is a corporate
`body.
`
`(see note (d))
`
`Patents Form 1/77
`
`Ex. 1085-0003
`
`

`

`Patents Form 1/77
`
`9.
`
`Enter the number of sheets for any of
`the following items you are filing with
`this form. Do not count copies of the
`same document
`
`Continuation sheets of this form
`
`Description 22
`
`Claim(s) 2
`
`Abstract
`
`Drawing(s)
`
`10.
`
`If you are also filing any of the
`following, state how many against each
`item.
`
`Priority documents
`
`Translations of priority documents
`
`Statement of inventorship and right
`to grant of a patent (Patents Form
`7/77)
`
`Request for preliminary examination
`and search (Patents Form 9/77)
`
`Request for substantive examination
`(Patents Form 10/77)
`
`Any other documents
`(please specify)
`
`11.
`
`I/We request the grant of a patent on the basis of this
`application
`
`Signature
`
`Date
`
`12. Name and daytime telephone number
`of person to contact in the United
`Kingdom
`
`B.A. Yorke & Co.
`Mrs. E. Cheetham
`020 8560 5847
`
`17 October 2001
`
`Warning
`After an application for a patent has been filed. the Comptroller of the Patent Office will consider whether
`publication or communication of the invention should be prohibited or restricted under Section 22 of the Patents
`Act 1977. You will be informed if it is necessary to prohibit or restrict your invention in this way. Furthermore, if
`you live in the United Kingdom, Section 23 of the Patents Act 1977 stops you from applying for a patent abroad
`without first getting written permission from the Patent Office unless an application has been filed at least 6
`weeks beforehand in the united Kingdom for a patent for the same invention and either no direction prohibiting
`publication or communication has been given, or any such direction has been revoked.
`Notes
`a)
`If you need help to fill in this form or you have any questions, please contact the Patent Office on 0645
`500505.
`
`b)
`
`c)
`
`d)
`e)
`
`Write your answers in capital letters using black ink or you may type them.
`If there is not enough space for all the relevant details on any part of this form, please continue on a
`separate sheet of paper and write 'see continuation sheet" in the relevant part(s). Any continuation sheet
`should be attached to this form.
`Once you have filled in the form you must remember to sign and date it.
`For details of the fee and ways to pay please contact the Patent Office.
`
`Patents Form 1/77
`
`Ex. 1085-0004
`
`

`

`Case 4-31671 P3
`
`Organic Compounds
`
`The.present invention relates to a new use, in particular a new use for a compound group
`comprising derivatives of rapamycin.
`
`Suitable derivatives of rapamycin include e.g. compounds of formula I
`
`wherein
`R1 is CH3 or C3_6alkynyl,
`R2 is H or -CH 2-CH 2-OH,
`X is =0 or (H,OH)
`provided that R2 is -CH 2-CH2-OH when R, is CH3 and X is =0.
`
`Most of the compounds of formula I are either generically or specifically disclosed in WO
`94/09010, WO 95/16691 or WO 96/41807, which are incorporated herein by reference, the
`compound of formula I wherein X is =0, R, is 2-pentynyl and R2 is -CH 2-CH 2-OH being novel
`and forming part of the invention.
`
`A preferred compound is 40-0-(2-hydroxyethyl)-rapamycin (referred thereafter as
`Compound A), disclosed as Example 8 in WO 94/09010.
`
`Certain compounds of formula I have, on the basis of observed activity, e.g. binding to
`macrophilin-12 (also known as FK-506 binding protein or FKBP-1 2), e.g. as described in
`WO 94/09010, WO 95/16691 or WO 96/41807, been found to be useful e.g. as
`immunosuppressant, e.g. in the treatment of acute allograft rejection. It has now been found
`that Compounds of formula I have potent antiproliferative properties which make them
`
`Ex. 1085-0005
`
`

`

`Case 4-31671 P3
`
`-2-
`
`4
`
`useful for cancer chemotherapy, particularly of solid tumors, especially of advanced solid
`tumors. There is still the need to expand the armamentarium of cancer treatment of solid
`tumors, especially in cases where treatment with anticancer compounds is not associated
`with disease regression or stabilization.
`
`In accordance with the particular findings of the present invention, there is provided:
`1.1 A method for treating solid tumors in a subject in need thereof, comprising
`administering to said subject a therapeutically effective amount of a compound of
`formula I.
`1.2 A method for inhibiting growth of solid tumors in a subject in need thereof, comprising
`administering to said subject a therapeutically effective amount of a compound of
`formula I.
`1.3 A method for inducing tumor regression, e.g. tumor mass reduction, in a subject in
`need thereof, comprising administering to said subject a therapeutically effective
`amount of a compound of formula I.
`1.4 A method for treating solid tumor invasiveness or symptoms associated with such
`tumor growth in a subject in need thereof, comprising administering to said subject a
`therapeutically effective amount of a compound of formula I.
`1.5 A method for preventing metastatic spread of tumours or for preventing or inhibiting
`growth of micrometastasis in a subject in need thereof, comprising administering to
`said subject a therapeutically effective amount of a compound of formula I.
`1.6 A method for the treatment of a disease associated with deregulated angiogenesis in
`a subject in need thereof, comprising administering to said subject a therapeutically
`effective amount of a compound of formula I.
`1.7 A method for inhibiting or controlling deregulated angiogenesis in a subject in need
`thereof, comprising administering to said subject a therapeutically effective amount of
`a compound of formula I.
`1.8 A method for enhancing the activity of an anticancer agent or for overcoming
`resistance to an anticancer agent in a subject in need thereof, comprising
`administering to said subject a therapeutically effective amount of a compound of
`formula I either concomitantly or sequentially with said anticancer agent.
`1.9 A method according to 1.8 wherein the anticancer agent is an inhibitor of signal
`transduction pathways directed either against host cells or processes involved in tumor
`
`Ex. 1085-0006
`
`

`

`Case 4-31671P3 .
`
`-3-
`
`formation and/or metastases formation or utilised by tumor cells for proliferation,
`
`survival, differentiation or development of drug resistance.
`
`1.10 A method as indicated above, wherein the compound of formula I is administered
`
`intermittently.
`
`By"solid tumors" are meant tumors and/or metastasis (whereever located) other than
`
`lymphatic cancer, e.g. pancreatic tumors; melanomas; lung cancer, e.g. small cell lung
`
`cancer; breast cancer; epidermoid carcinomas; renal-cell carcinomas; neuroendocrine
`
`tumors; genitourinary cancer, e.g. cervical, uterine, ovarian, prostate or bladder cancer;
`
`gastrointestinal cancer, e.g. gastric or colon cancer; glioblastomas; head and/or neck
`
`cancer; soft-tissue sarcomas.
`
`Where hereinbefore and subsequently a tumor, a tumor disease, a carcinoma or a cancer is
`
`mentioned, also metastasis in the original organ or tissue and/or in any other location are
`implied alternatively or in addition, whatever the location of the tumor and/or metastasis is.
`
`Examples of diseases associated with deregulated angiogenesis include e.g. neoplastic
`
`diseases, e.g. solid tumors.
`
`Angiogenesis is regarded as a prerequisite for those tumors which grow beyond a certain
`
`diameter, e.g. about 1-2 mm.
`
`2.
`
`As alternative to the above the present invention also provides:
`A compound of formula I wherein X is =0, R1 is 2-pentynyl and R2 is -CH 2-CH 2-OH for
`use as a pharmaceutical;
`
`3.
`
`4.
`
`5.
`
`6.
`
`A compound of formula I for use in any method as defined under 1.1 to 1.10 above;
`
`A compound of formula I for use in the preparation of a pharmaceutical composition
`
`for use in any method as defined under 1.1. to 1.10. above;
`A pharmaceutical composition comprising a compound of formula I wherein X is =0,
`R1 is 2-pentynyl and R2 is -CH 2-CH 2-OH together with one or more pharmaceutically
`acceptable diluents or carriers therefor;
`A pharmaceutical composition for use in any method as defined under 1.1 to 1.10
`
`above comprising a compound of formula I together with one or more pharmaceutically
`
`acceptable diluents or carriers therefor.
`
`Ex. 1085-0007
`
`

`

`Case 4-31671 P3
`
`-4-
`
`The compounds of formula I also defined as mTOR (mammalian "Target of Rapamycin")
`
`inhibitors may be administered as the sole active ingredient or together with other
`chemotherapeutic drugs. In a series of further specific or alternative embodiments, the
`
`present invention also provides:
`
`7.
`
`8.
`
`9.
`
`A pharmaceutical combination comprising a) a first agent which is a mTOR inhibitor,
`e.g. a macrocyclic lactone, e.g. rapamycin or a derivative thereof, e.g. CC1779 or a
`compound of formula I, e.g. Compound A, and b) a co-agent which is a
`chemotherapeutic agent, e.g. as defined hereinafter.
`A method as defined above comprising co-administration, e.g. concomitantly or in
`
`sequence, of a therapeutically effective amount of a mTOR inhibitor, e.g. a
`macrocyclic lactone, e.g. rapamycin or a derivative thereof, e.g. CC1779 or a
`compound of formula I, e.g. Compound A, and a second drug substance, said second
`drug substance being a chemotherapeutic agent, e.g. as indicated hereinafter.
`A method for treating a lymphatic cancer, e.g. for treating tumor invasiveness or
`
`symptoms associated with such tumor growth in a subject in need thereof, comprising
`co-administering to said subject, e.g. concomitantly or in sequence, of a
`therapeutically effective amount of a mTOR inhibitor, e.g. a macrocyclic lactone, e.g.
`rapamycin or a derivative thereof, e.g. CC1779 or a compound of formula I, e.g.
`Compound A, and a second drug substance, said second drug substance being a
`chemotherapeutic agent, e.g. as indicated hereinafter.
`
`CC1779 is an 40-ester of rapamycin with 2,2-bis(hydroxymethyl)propionic acid or a
`pharmaceutically acceptable salt thereof, and is disclosed e.g. in USP 5,362,718, the
`content thereof being incorporated herein by reference.
`
`By "lymphatic cancer" are meant e.g. lymphomas and lymphotic leukemias. The lymphocytic
`
`leukemias include disorders such as acute and chronic lymphocytic leukemias. Lymphomas
`encompass a wide variety of cancers characterized by lymphocyte proliferation, for example
`Hodgkin's lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, AIDS-related
`
`lymphomas, diffuse large cell lymphoma, T-cell lymphoma or cutaneous T-cell lymphoma.
`
`Ex. 1085-0008
`
`

`

`Case 4-31671 P3
`
`-5-
`
`By the term "other chemotherapeutic", there is meant especially any chemotherapeutic
`other than a compound of formula I. It includes but is not limited to, an aromatase inhibitor,
`an antiestrogen, an anti-androgen (especially in the case of prostate cancer), a gonadorelin
`agonist, a topoisomerase I inhibitor, a topoisomerase II inhibitor, a microtubule active agent,
`an alkylating agent, an antineoplastic antimetabolite, a platin compound, a compound
`targeting/decreasing a protein kinase activity and further anti-angiogenic compound, a
`bradykinin 1 receptor, an angiotensin II antagonist, a cyclooxygenase inhibitor, a matrix
`metalloproteinase inhibitor, a bisphosphonate, a methionine aminopeptidase inhibitor, e.g.
`bengamide or a derivative thereof, a histone deacetylase inhibitor, a heparanase inhibitor
`(prevents heparan sulphate degradation), e.g. PI-88, a proteosome inhibitor, e.g. PS-341, a
`biological response modifier, preferably a lymphokine or interferons, or any epitholone or
`derivative thereof, e.g. epitholone B.
`
`The term "aromatase inhibitor" as used herein relates to a compound which inhibits the
`estrogen production, i.e. the conversion of the substrates androstenedione and testo-
`sterone to estrone and estradiol, respectively. The term includes, but is not limited to
`steroids, especially exemestane and formestane and, in particular, non-steroids, especially
`aminoglutethimide, vorozole, fadrozole, anastrozole and letrozole. Exemestane can be
`administered, e.g., in the form as it is marketed, e.g. under the trademark AROMASINTM.
`Formestane can be administered, e.g., in the form as it is marketed, e.g. under the
`trademark LENTARON T . Fadrozole can be administered, e.g., in the form as it is marketed,
`e.g. under the trademark AFEMATM. Anastrozole can be administered, e.g., in the form as it
`is marketed, e.g. under the trademark ARIMIDEX M . Letrozole can be administered, e.g., in
`the form as it is marketed, e.g. under the trademark FEMARA TM or FEMAR TM .
`Aminoglutethimide can be administered, e.g., in the form as it is marketed, e.g. under the
`trademark ORIMETEN TM . A combination of the invention comprising a chemotherapeutic
`agent which is an aromatase inhibitor is particularly useful for the treatment of hormone
`receptor positive breast tumors.
`
`The term "antiestrogen" as used herein relates to a compound which antagonizes the effect
`of estrogens at the estrogen receptor level. The term includes, but is not limited to
`tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen can be
`administered, e.g., in the form as it is marketed, e.g. under the trademark NOLVADEXTM.
`
`Ex. 1085-0009
`
`

`

`Case 4-31671 P3
`
`Raloxifene hydrochloride can be administered, e.g., in the form as it is marketed, e.g. under
`the trademark EVISTATM. Fulvestrant can be formulated as disclosed in US 4,659,516 or it
`can be administered, e.g., in the form as it is marketed, e.g. under the trademark
`
`FASLODEXTM .
`
`The term "topoisomerase I inhibitor" as used herein includes, but is not limited to topotecan,
`irinotecan, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-
`166148 (compound Al in W099/17804). Irinotecan can be administered, e.g., in the form
`as it is marketed, e.g. under the trademark CAMPTOSAR TM . Topotecan can be
`
`administered, e.g., in the form as it is marketed, e.g. under the trademark HYCAMTINTM.
`
`The term "topoisomerase II inhibitor" as used herein includes, but is not limited to the
`
`antracyclines such as doxorubicin (including liposomal formulation, e.g. CAELYXTM),
`epirubicin, idarubicin and nemorubicin, the anthraquinones mitoxantrone and losoxantrone,
`and the podophillotoxines etoposide and teniposide. Etoposide can be administered, e.g., in
`
`the form as it is marketed, e.g. under the trademark ETOPOPHOSTM. Teniposide can be
`administered, e.g., in the form as it is marketed, e.g. under the trademark VM 26-BRISTOL
`TM. Doxorubicin can be administered, e.g., in the form as it is marketed, e.g. under the
`
`trademark ADRIBLASTIN TM . Epirubicin can be administered, e.g., in the form as it is mar-
`
`keted, e.g. under the trademark FARMORUBICINTM. Idarubicin can be administered, e.g., in
`
`the form as it is marketed, e.g. under the trademark ZAVEDOS TM . Mitoxantrone can be
`
`administered, e.g., in the form as it is marketed, e.g. under the trademark NOVANTRONTM.
`
`The term "microtubule active agent" relates to microtubule stabilizing and microtubule
`destabilizing agents including, but not limited to taxanes, e.g. paclitaxel and docetaxel,
`vinca alkaloids, e.g., vinblastine, especially vinblastine sulfate, vincristine especially
`vincristine sulfate, and vinorelbine, discodermolide and epothilones and derivatives thereof.
`Docetaxel can be administered, e.g., in the form as it is marketed, e.g. under the trademark
`
`TAXOTERETM. Vinblastine sulfate can be administered, e.g., in the form as it is marketed,
`
`e.g. under the trademark VINBLASTIN R.P.TM. Vincristine sulfate can be administered, e.g.,
`in the form as it is marketed, e.g. under the trademark FARMISTINTM. Discodermolide can
`
`be obtained, e.g., as disclosed in US 5,010,099.
`
`Ex. 1085-0010
`
`

`

`Case 4-31671 P3
`
`-7-
`
`The term "alkylating agent" as used herein includes, but is not limited to cyclophosphamide,
`ifosfamide and melphalan. Cyclophosphamide can be administered, e.g., in the form as it is
`
`marketed, e.g. under the trademark CYCLOSTINTM. Ifosfamide can be administered, e.g., in
`
`the form as it is marketed, e.g. under the trademark HOLOXANTM.
`
`The term "antineoplastic antimetabolite" includes, but is not limited to 5-fluorouracil,
`
`capecitabine, gemcitabine, methotrexate and edatrexate. Capecitabine can be
`
`administered, e.g., in the form as it is marketed, e.g. under the trademark XELODATM.
`
`Gemcitabine can be administered, e.g., in the form as it is marketed, e.g. under the
`
`trademark GEMZAR TM .
`
`The term "platin compound" as used herein includes, but is not limited to carboplatin, cis-
`platin and oxaliplatin. Carboplatin can be administered, e.g., in the form as it is marketed,
`
`e.g. under the trademark CARBOPLATTM. Oxaliplatin can be administered, e.g., in the form
`
`as it is marketed, e.g. under the trademark ELOXATINTM.
`
`The term "compounds targeting/decreasing a protein kinase activity and further anti-
`angiogenic compounds" as used herein includes, but is not limited to protein tyrosine kinase
`and/or serine and/or threonine kinase inhibitors, e.g. compounds targeting, decreasing or
`inhibiting the activity of the epidermal growth factor family of receptor tyrosine kinases
`(EGFR, ErbB2, ErbB3, ErbB4), the vascular endothelial growth factor family of receptor
`tyrosine kinases (VEGFR), the platelet-derived growth factor-receptors (PDGFR), the
`insulin-like growth factor receptor 1 (IGFR1), members of the c-Abl family and their gene-
`fusion products (e.g. BCR-Abl) and/or members of the protein kinase C (PKC) and Raf
`family of serine/threonine kinases, anti-angiogenic compounds having another mechanism
`
`for their activity.
`
`Compounds which target, decrease or inhibit the activity of VEGFR are especially
`compounds which inhibit the VEGF receptor tyrosine kinase, compounds which inhibit a
`VEGF receptor and compounds binding to VEGF, and are in particular those compounds,
`
`proteins and monoclonal antibodies generically and specifically disclosed in WO 98/35958,
`e.g. 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt
`thereof, e.g. the succinate, or in WO 00/09495, WO 00/27820, WO 00/59509, WO
`
`Ex. 1085-0011
`
`

`

`Case 4-31671 P3
`
`-8-
`
`9
`
`98/11223, WO 00/27819 and EP 0769 947; those as described by M. Prewett et al in
`Cancer Research 59 (1999) 5209-5218, by F. Yuan et al in Proc. NatI. Acad. Sci. USA, vol.
`93, pp. 14765-14770, Dec. 1996, by Z. Zhu et al in Cancer Res. 58, 1998, 3209-3214, and
`
`by J. Mordenti et al in Toxicologic Pathology, Vol. 27, no. 1, pp 14-21, 1999; in WO
`00/37502 and WO 94/10202; Angiostatin TM , described by M. S. O'Reilly et al, Cell 79,1994,
`315-328; EndostatinTM, described by M. S. O'Reilly et al, Cell 88, 1997, 277-285; anthranilic
`
`acid amides; ZD4190; ZD6474; SU5416; SU6668; or anti-VEGF antibodies or anti-VEGF
`receptor antibodies,e.g. RhuMab;
`
`compounds which target, decrease or inhibit the activity of the epidermal growth factor
`receptor family are especially compounds which inhibit the EGF receptor tyrosine kinase,
`compounds which inhibit the EGF receptor or ErbB2 and compounds binding to EGF, and
`are in particular those compounds generically and specifically disclosed in WO 97/02266,
`
`e.g. the compound of ex. 39, or in EP 0 564 409, WO 99/03854, EP 0520722, EP 0 566
`226, EP 0 787 722, EP 0 837 063, US 5,747,498, WO 98/10767, WO 97/30034, WO
`97/49688, WO 97/38983 and, especially, WO 96/33980; e.g. trastuzumab (HerpetinR),
`cetuximab, Iressa, OSI-774, CI-1033, EKB-569 or GW-2016;
`
`compounds which target, decrease or inhibit the activity of PDGFR are especially
`compounds which inhibit the PDGF receptor, e.g. a N-phenyl-2-pyrimidine-amine derivative,
`
`e.g. imatinib;
`
`compounds which target, decrease or inhibit the activity of c-Abl family members and their
`
`gene fusion products, e.g. a N-phenyl-2-pyrimidine-amine derivative, e.g. imatinib;
`PD180970; AG957; or NSC 680410;
`
`compounds which target, decrease or inhibit the activity of protein kinase C and Raf family
`
`members are especially those staurosporine derivatives disclosed in EP 0 296 110, e.g.
`
`midostaurin; examples of further compounds include e.g. UCN-01, safingol, BAY 43-9006,
`Bryostatin 1, Perifosine; Ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521; or
`
`LY333531/LY379196;
`
`Further anti-angiogenic compounds are e.g. thalidomide (THALOMID) and TNP-470.
`
`Ex. 1085-0012
`
`

`

`Case 4-31671 P3
`
`-9-
`
`The term cyclooxygenase inhibitor as used herein includes, but is not limited to, e.g.
`
`celecoxib (CelebrexR), rofecoxib (VioxxR), etoricoxib, valdecoxib or a 5-alkyl-2-
`arylaminophenylacetic acid, e.g. 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenyl acetic acid.
`
`The term "matrix metalloproteinase inhibitor" as used herein includes, but is not limited to
`
`TAA211 or AAJ996.
`
`The term "histone deacetylase inhibitor" as used herein includes, but is not limited to MS-
`
`27-275, SAHA, pyroxamide, FR-901228 or valproic acid.
`
`The term "gonadorelin agonist" as used herein includes, but is not limited to abarelix,
`goserelin and goserelin acetate. Goserelin is disclosed in US 4,100,274 and can be
`
`administered, e.g., in the form as it is marketed, e.g. under the trademark ZOLADEX TM .
`Abarelix can be formulated, eg. as disclosed in US 5,843,901.
`
`The term "anti-androgens" as used herein includes, but is not limited to, bicalutamide
`(CASODEXTM ), which can be formulated, e.g. as disclosed in US 4,636,505.
`
`The term "bisphosphonates" as used herein includes, but is not limited to, etridonic acid,
`clodronic acid, tiludronic acid, pamidronic acid, alendronic acid, ibandronic acid, risedronic
`acid and zoledronic acid. "Etridonic acid" can be administered, e.g., in the form as it is
`
`marketed, e.g. under the trademark DIDRONELTM. "Clodronic acid" can be administered,
`
`e.g., in the form as it is marketed, e.g. under the trademark BONEFOSTM. "Tiludronic acid"
`
`can be administered, e.g., in the form as it is marketed, e.g. under the trademark SKELIDTM.
`"Pamidronic acid" can be administered, e.g., in the form as it is marketed, e.g. under the
`
`trademark AREDIATM. "Alendronic acid" can be administered, e.g., in the form as it is
`
`marketed, e.g. under the trademark FOSAMAXTM. "Ibandronic acid" can be administered,
`
`e.g., in the form as it is marketed, e.g. under the trademark BONDRANATTM. "Risedronic
`
`acid" can be administered, e.g., in the form as it is marketed, e.g. under the trademark
`
`ACTONELTM. "Zoledronic acid" can be administered, e.g., in the form as it is marketed, e.g.
`under the trademark ZOMETATM.
`
`Ex. 1085-0013
`
`

`

`Case 4-31671 P3
`
`-10-
`
`I
`
`In each case where citations of patent applications or scientific publications are given, the
`subject-matter relating to the compounds is hereby incorporated into the present application
`by reference. Comprised are likewise the pharmaceutically acceptable salts thereof, the
`corresponding racemates, diastereoisomers, enantiomers, tautomers as well as the
`corresponding crystal modifications of above disclosed compounds where present, e.g.
`solvates, hydrates and polymorphs, which are disclosed therein. The compounds used as
`active ingredients in the combinations of the invention can be prepared and administered as
`described in the cited documents, respectively. Also within the scope of this invention is the
`combination of more than two separate active ingredients as set forth above, i.e. a
`pharmaceutical combination within the scope of this invention could include three active
`ingredients or more. Further both the first agent and the co-agent are not the identical
`ingredient.
`
`Utility of the compounds of formula I in treating solid tumors as hereinabove specified, may
`be demonstrated in animal test methods as well as in clinic, for example in accordance with
`the methods hereinafter described.
`A.
`In Vitro
`A.1 Antiproliferative activity in combination with other agents
`A cell line, e.g. KB-31, A549 or HCT1 16, is added to 96-well plates (1,500 cells/well in
`100 pl medium) and incubated for 24 hr. Subsequently, a two-fold dilution series of each
`compound (Compound of formula I or a known anticancer agent) is made in separate tubes
`(starting at 8 x the IC50 of each compound) either alone or in paired combinations, and the
`dilutions are added to the wells. The cells are then re-incubated for 3 days. Methylene blue
`staining is performed on day 4 and the amount of bound dye (proportional to the number of
`surviving cells that bind the dye) determined. IC50s are subsequently determined using the
`Calcusyn program, which provides a measure of the interaction, namely the so-called non-
`exclusive combination index (CI), where: Cl - 1 = the interaction is nearly additive; 0.85 -
`0.9 = slight synergism; < 0.85 = synergy. In this assay, the compounds of formula I show
`interesting antiproliferative activity in combination with other anticancer agent. For example
`following CI values are obtained with a combination of Compound A and cisplatinum,
`paclitaxel, gemcitabine and doxorubicin.
`
`Cl
`
`Cell line Cisplatinum
`
`Paclitaxel
`
`Gemcitabine Doxorubicin
`
`Ex. 1085-0014
`
`

`

`Case 4-31671 P3
`
`-11
`
`-
`
`0.9
`
`0.74
`
`0.3
`
`0.79
`
`0.76
`
`0.9
`
`0.7
`
`0.64
`
`0.52
`
`KB-31
`
`A549
`
`HCT116
`
`0.74
`
`0.47
`
`0.47
`
`A.2 Antiangiogenic activity
`In vitro assay of the antiproliferative activity of a compound of formula I, e.g. Compound
`
`A against human umbilical vein endothelial cells (HUVECs) demonstrates IC50 values of 120
`± 22 pM and 841 + 396, and > 10 000 pM for VEGF- and bFGF- and FBS-stimulated
`
`proliferation, respectively. Additionally, no significant effects of Compound A on bFGF-
`
`stimulated normal human dermal fibroblast (NHDF) proliferation are observed over the
`
`same concentration range. These results indicate that Compound A inhibits the proliferation
`
`of HUVECs, being particularly potent against the VEGF-induced proliferation, VEGF being a
`
`key pro-angiogenic factor.
`
`B.
`
`In Vivo
`
`B.1 Activity in A549 human lung tumor xenografts
`
`Fragments of A549 tumors (approx. 25 mg; Cell line CCL 185, ATCC, Rockville MD,
`USA), a cell line shown to be sensitive to Compound A in vitro, are transplanted
`
`subcutaneously into the left flank of BALB/c nude mice. Treatment is started on day 7 or
`day 12 following tumor transplantation. The compound to be tested is administered p.o.
`
`once per day from day 7/12 to day 38/55, respectively. Antitumor activity is expressed as
`
`T/C% (mean increase in tumor volumes of treated animals divided by the mean increase of
`
`tumor volumes of control animals multiplied by 100) and % regressions (tumor volume
`
`minus initial tumor volume divided by the initial tumor volume and multiplied by 100). In this
`
`assay, when administered at a daily dose ranging from 0.1 mg/kg to 2.5 mg/kg, the
`
`compounds of formula I exhibited dose-dependent inhibition of tumor growth; for example in
`
`one representative experiment Compound A when administered at a dose of 2.5 mg/kg
`
`results in persisting regressions (41 %); a dose of 0.5 mg/kg results in transient regressions
`
`(38 % on day 17), with a final T/C of 16 %, and a dose of 0.1 mg/kg slows tumor growth
`
`resulting in a final T/C of 43 % (T/C for control animals is 100%).
`
`Ex. 1085-0015
`
`

`

`Case 4-31671 P3
`
`-12-
`
`B.2 Activity in KB-31 human epidermoid tumor xenografts
`
`Fragments of KB-31 tumors (approx. 25 mg; Roswell Park Memorial Institute Buffalo,
`
`NY, USA), a cell line shown to be comparatively resistant to Compound A in vitro, are
`transplanted subcutaneously into the left flank of BALB/c nude mice. Treatment is started
`on day 7 or on day 10 following tumor transplantation. The compound to be tested is
`administered p.o. once per day from day 7/10 to day 25/35, respectively. Antitumor activity
`
`is expressed as T/C% as indicated above. In this assay, when administered at a daily dose
`ranging from 0.5 mg/kg to 2.5 mg/kg, the compounds of formula I inhibit tumor growth; for
`example in one representative experiment Compound A when administered at a dose of 2,5
`mg/kg/day results in a final T/C cvalue of 25%(T/C for control animals is 100%).
`
`B.3 Activity in CA20948 rat pancreatic tumors
`Tumors are established in male Lewis rats by subcutaneous injection of CA20948
`
`tumor cell suspension derived from donor rats into the left flank. Treatment is started on day
`4 post inoculation. The compound to be tested is administered p.o. once per day (6 days a
`week) from day 4 to day 9-15 post inoculation. Antitumor activity is expressed as T/C% as
`indicated above. In this assay, when administered at a daily dose of 0.5 mg/kg to 2.5 mg/kg,
`the compounds of formula I inhibit tumor growth; for example in a representative experiment
`Compound A when administered p.o. at a daily dose of 2.5 mg/kg results in a final T/C value
`of 23 %. In the same experiment, intermittent administration of Compound A, 5mg/kg twice
`
`per week, results in a final T/C value of 32%. Compound A significantly and consistently
`decreases in these assays the rate of CA20948 pancreatic tumor growth when compared to
`vehicle controls (T/C for control animals is defined as 100%).
`
`Compound A has been tested in further tumor models in accordance with the procedure as
`
`disclosed above. For example, a daily dosage of 2.5 mg/kg or 5 mg/kg Compound A
`produced final T/Cs of 18% and 9% when administered to the human NCI H-596 lung tumor
`
`model and the human MEXF 989 melanoma tumor model, respectively, 5 mg/kg produced
`final T/Cs of 20% (primary tumor) and 36% (cervical lymph node metastases) when
`administered to the orthotopic mouse B1 6/BL6 melanoma tumor model and 24% when
`administered to the human AR42J pancreatic tumor model, 2.5 mg/kg produced a final T/C
`of 28%