UNITED STATES DEPARTMENT OF COMMERCE
`United States Patent and Trademark Office
`
`September 09, 2015
`
`THIS IS TO CERTIFY THAT ANNEXED IS A TRUE COPY FROM THE
`RECORDS OF THIS OFFICE OF THE FILE WRAPPER AND CONTENTS
`OF:
`
`APPLICATION NUMBER: 081416,673
`FILING DATE: April 07, 1995
`PATENT NUMBER: 5,665, 772
`ISSUE DATE: September 09, 1997
`
`By Authority of the
`Under Secretary of Commerce for Intellectual Property
`and Director of the United States Patent and Trademark Office
`
`jYJ ~
`
`M. TARVER
`Certifying Officer
`
`Breckenridge Exhibit 1002
`Breckenridge v. Novartis AG
`
`

`
`'~ 'y.
`
`- (-..---...--'.,'""";'I'. ----~#lll'!J.111'!'-""1C'¥(tt"'l ......
`!lllllll"W®_,,, m,
`
`·"
`·-~94109010 ~ ~ced h ~II/+ ~{Q,.'Sl/. PCT/EP93/02604
`{1__t c~{ tJ /) o1~199-S
`
`,
`
`....
`
`- 37 -
`
`CLAIMS
`
`(I)
`
`0 ,,,--
`
`24
`
`X is (H,H) or O;
`
`Y is (H,OH) or O;
`
`R 1 and R2 are independently selected from
`
`H, alkyl, thioalkyl. arylalkyl, hydroxyalkyl, dihydroxyalkyl,
`
`hydroxyalkylarylalkyl, dihydroxyalkylarylalky0alkoxyalkyl, acyloxyalkyl,
`\
`aminoalkyl, alkylaminoalkyl, alkoxycarbonylaminoalkyl, acylaminoalkyl,
`
`

`
`,_
`.. -· WD 94/09010
`
`•
`
`-38-
`
`• PCT/EP93/02604
`
`arylsulfonamidoalkyl, allyl. clihydroxyalkylallyl, dioxolanylallyl,
`caralkoxyalkyl, and (R.3hSi where each R3 is independently selected from ~
`methyl, ethyl. isopropyl. ,!-butyl. and phenyl; wherein "alk-" or "alkyl" refers
`to cl-6 alkyl, branched or linear, preferably c;.3 alkyl, in which the carbon
`chain may be optionally interrupted by an ether (-0-) linkage; and
`
`R' is m thy! or R" and R 1 together fOim ~ alkylene;
`
`provided th R ! and R: are not both H; and
`pro~ided that here R1 is carbalkoxyalkyl or (Ri),Si. X and Y are not both 0.
`
`2.
`
`Compounds ace
`
`· g to claim 1 selected from the following:
`
`1.
`2.
`3.
`4.
`5.
`6.
`
`7.
`
`8.
`
`9.
`
`40-0-Allyl-rapam cin
`40-0-[3'-(2.2-Dim yl-l.3-dioxolan-4{S)-yl)-prop-2'-cn-l'-yl]-rapamycin
`'-Dihydroxypent-2'-en-1 '-yl)-rapamycin
`
`(2'E. 4'S)-40-0-(4'
`
`40-0-(2-Hydroxy)etbo carbonylmethyl-rapamycin
`
`40-0-(2-Hydroxy)cthyl-
`
`40-0-(3-Hydroxy)propyl-
`
`amycin
`
`10. ~0-(6-Hydroxy)hexyl-rap ycin
`
`11. 40-0-[2-(2-Hydroxy)ethoxy]c yl-rapamycin
`12. 4Q.0-[(3S)-2.2-Dimethyldioxol
`-3-yl]methyl-rapamycin
`13. ~0-[(25)-2,3-Dihydroxypiop-l- J-rapamycin
`
`14. 40-0-(2-Acetoxy)ethyl-rapamycin
`
`15. 40-0-(2-Nicotinoyloxy)cthyl-rapamycin
`
`16. ~0-[2-(N-Morpholino )acetoxy]ethyl-~amycin
`
`\
`
`

`
`' )."
`
`"'- """',
`· wd 9410901 o
`. .......;~ __
`
`•
`
`•
`
`PCT/EP93/02604
`
`- 39 -
`
`~\
`17. ~6-0-(2-N-Imidazolylacemxy)ethyl-rapamycin
`18. 40-0-[2-(N-Methyl-N '-piperazinyl)acetoxy ]ethyl-rapamycin
`3~f-Desmethyl-39,40-0,0-ethylene-rapamycin
`
`19.
`
`20.
`(26~~26-Dihydro-40-0-(2-hydroxy)ethyl-rapamycin
`21. 28-0-
`ethyl-rapamycin
`22. 40-0-(2
`·noethyl)-rapamycin
`
`23.
`
`40-0-(2-~cetaminoethyl)-rapamycin
`
`40-0-(2-N~~·namidoethyl)-rapamycin
`24.
`25. 40-0-(2-(N-
`ethyl-imidazo-2 '-ylcarbethoxamido )ethyl)-rapamycin
`
`26.
`
`40-0-(2-Ethox carbonylaminoethyl)-rapamycin
`
`27. 40-0-(2-Tolyls~fon~doethy 1)-rapamycin
`40-0-[2-( 4 ',5 '-D~\oethoxy-1 ',2' ,3 '-triazol-1 '-y 1)-eth yl )-rapamycin
`
`28.
`
`3.
`
`4.
`
`5.
`
`6.
`
`7.
`
`Compounds according to clallµ 1 where X and Y are both 0, R2 is H, R4 is methyl,
`and R 1 is other than H.
`
`\
`
`.
`
`40-0-(2-Hydroxy)ethyl-rapamy\
`
`Compounds according to any one ~~!aims 1 through 4 obtained or obtainable by (i)
`
`reacting a rapamycin, deoxorapam~~\· or dihydrorapamycin (optionally in 0-
`protected form) with an organic radic' attached to a leaving group under suitable
`acidic or neutral reaction conditions, an
`(ii) optionally reducing the product..
`
`A compound according to any one of clai
`
`1-5 for use as a pharmaceutical.
`
`A pharmaceutical composition comprising a co\pound according to any one of
`
`claims 1-5 together with a pharmaceutically acceP.table diluent or carrier.
`
`8.
`
`Use of a compound according to claims 1-5 in th\ufacture of a medicament for
`
`

`
`r~>~
`~
`w094t09010
`
`PCT /EP93/02604
`
`•
`
`- 40 -
`
`or preventing any of the following conditions:
`
`(i)
`
`(ii)
`
`(iii)
`
`(iv) asthma.
`
`(v)
`
`..
`
`(vi) tumors or hyp
`
`roliferative disorders, or
`
`(vii) fungal infections,
`
`(viii) inflammation,
`
`having Mip or Mip-like factors, or
`
`9.
`
`Novel products, processes, and utilities substantially as described herein.
`
`
`
`

`
`\W
`
`• ',-~,
`
`PCf
`INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY {PC1)
`
`WORLD INTELLECTUAL PROPERTY ORGANIZATION'
`International Bureau
`'
`
`(51) International Patent Oassification 5 :
`C07D 498/18, C07F 7 /18
`A61K 31/435 // C07D 498/18
`C07D 311 :00, 273:00, 221 :00
`
`(11) International Publication Number:
`
`WO 94/09010
`
`Al
`
`(43) International Publication Date:
`
`28 April 1994 (28.04.94)
`
`(21) International Application Number:
`
`PCT /EP93/02604
`
`(22) International Filing Date:
`
`24 September 1993 (24.09.93)
`
`(72) Inventors; and
`(75) Inventors/ Applicants {for US only) : COTTENS, Sylvain
`[CH/CH]; In den Reben 12, CH-4108 Witterswil (CH).
`SEDRANI, Richard [LU/CH]; Herrengrabenweg 15,
`CH-4054 Basie (CH).
`
`(30) Priority data:
`9221220.8
`
`9 October I 992 (09. I 0.92)
`
`GB
`
`(74)Common Representative: SANDOZ LTD.; Patents &
`Trademarks Div., Lichtstrasse 35, CH-4002 Basie {CH).
`
`(71) Applicant {for AT only): SANDOZ-ERFINDUNGEN
`VERWALTUNGSGESELLSCHAFT M.B.H. [AT/AT];
`Brunner Strasse 59, A-1230 Vienna (A 1).
`
`(71) Applicant {for DE only): SANDOZ-PATENT-GMBH [DE/ '
`DE); Humboldstrasse 3,_p-79539 Lorrach {DE).
`·
`
`(81) Designated States: AU, CA, CZ, FI, HU, JP, KR, NO, NZ,
`PL, RO, RU, SK, US, European patent {AT, BE, CH,
`DE, DK, ES, 'FR, GB, GR, IE, IT, LU, MC, NL, PT,
`SE).
`-
`
`(71) Applicant {for all designated States except AT DE US): SAN(cid:173)
`DOZ LTD. [CH/CH]; Lichtstrasse 35, CH-4002 Basie
`{CH).
`
`Published
`With international search report.
`
`(54)Title: 0-ALKYLATED RAPAMYCIN DERIVATIVES AND THEIR USE, PARTICULARLY AS IMMUNOSUPPRES(cid:173)
`SANTS
`
`(57) Abstract
`
`Novel 0-alkylated derivatives of rapamycin of for-
`mula (I), especially 40-0-alkylated derivatives, are found R •a
`to have pharmaceutical utility, particularly as immunosup(cid:173)
`pressants.
`
`so,,, .... ····
`~~a
`
`i J4
`
`1t
`
`0 _........
`
`13
`
`15
`
`20
`
`'/""
`19
`
`(I)
`
`24
`
`

`
`DECLARATION AND POWER OF ATTORNEY
`FOR UNITED STATES PATENT APPLICATION
`
`Case No.
`
`100-7932/PCT
`
`As a below named inventor, I hereby declare that:
`
`My residence, post office address and citizenship are as _stated below next to my name,
`
`and
`
`I believe that I am the original, first and sole inventor (if only one name is listed below)
`
`or an original, first and joint inventor (if more than one name is listed below) of the subject
`
`matter which is claimed and for which a United States patent is sought on the invention
`
`entitled
`
`0-ALKYLATED RAPAMYCIN DER_JVATIVES AND THEIR USE, PARTICULARLY
`AS IMMUNOSUPPRESSANTS
`
`the specification of which
`
`is attached hereto.
`
`was_ filed on
`
`19
`
`as application Serial No. 01
`
`and, if these brackets contain an X [ ], was amended on
`
`19
`
`[X] was filed as Patent Cooperation Treaty internation application No. PCT/EP93/02604
`
`on September 24
`
`, 19 9 3
`
`, if these brackets contain an X [ ], was
`
`amended under Patent Cooperation Treaty Article 19 on
`
`and, if these brackets contain an X [
`
`] , was amended on
`
`' 19
`
`' 19
`
`[ ] entered the national stage in the United States and was accorded Serial No.
`
`on
`
`was amended on
`
`' 19
`
`, and if these brackets contain an X [
`
`I hereby state thai. I have reviewed and understand the contents of the above-identified
`
`specifiction including the claims, as amended by any amendment(s) referred to above.
`
`I acknowledge my duty to disclose all information which is known by me to be material
`
`to patentability as defined in Title 37, Code of Federal Regulations, § 1.56.
`
`I hereby claim the benefit under Title 35, United States Code, §119 of any foreign
`
`application(s) for patent or inventor's certificate indicated below and of any Patent
`
`Cooperation Treaty international application(s) designating at least one country other than the
`
`United States indicated below and have also identified any foreign application(s) for
`
`- 1 -
`
`
`
`

`
`Case No. 100-7932/PCT
`
`patent or inventor's certificate and Patent Cooperation Treaty international application(s)
`
`designating at least one country other than the United States for the same subject matter and
`
`having a filing date before that of the application for said subject matter the priority of which
`
`is claimed:
`
`Country:
`
`Number:
`
`Filing Date:
`
`Priority Claimed:
`
`Great Britain
`
`9221220.8
`
`October 9, 1992
`
`[X] Yes[ ] No
`
`[' ] Yes[ ] No
`
`] Yes[ ] No
`
`[ ] Yes[ ] No
`
`[ ] Yes[ ] No
`
`[ ] Yes[ ] No
`
`I hereby claim the benefit under Title 35, United States Code, § 120 of any United
`
`States application(s) listed below and of any Patent Cooperation Treaty international
`
`application(s) designating the United States listed below and, insofar as the subject matter of
`
`each of the claims of this application is not disclosed in said prior application(s) in the
`
`manner required by the first paragraph of Title 35, United States Code, § 112, I acknowledge
`
`my duty to disclose all information known to me to be material to patentability as defined in
`
`Title 37. Code of Federal Regulations, § 1.56 which became available between the filing
`
`date(s) of the prior application(s) and the national or Patent Cooperation Treaty international
`
`filing date of this application:
`
`Application
`
`Serial No.
`none
`
`Status (Pending,
`
`Abandoned, Patented)
`
`- 2 -
`
`
`
`

`
`I hereby appoint the following:
`
`ROBERT S. HONOR
`THOMAS 0. MCGOVERN
`MELVYN M. KASSENOFF
`JOSEPH J. BOROVIAN
`DIANE E. FURMAN
`CARL W. BATTLE
`ANDREW N. PARFOMAK
`JOHN L. CHIATALAS
`CAROL A. LOESCHORN
`MICHAEL P. MORRIS
`THOMAS C. DOYLE
`
`Case No. 100-7932/PCT
`
`~g,_No. 22.801
`Reg. No. 25,741
`Reg. Nc>--26;389-
`Reg. No. 2.6.63-1-
`Re~-31-.-1.04-
`.Reg. No. 3Q73J-
`Reg; No. 32 431
`Reg. No. 31 ,818
`._Reg No 35,590 -
`Reg. No. 34 513_
`-Reg. No. 22,340
`
`/ /
`
`respectively and individually, as my attorneys and/or agents, with full power of substitution
`
`and revocation, to prosecute this application and to transact all business in the Patent and
`
`Trademarks Office connected therewith. Please address all communications to ROBERT S.
`
`HONOR, SANDOZ CORPORATION, 59 Route 10, East Hanover, New Jersey 07936-1080,
`
`whose telephone number is 201-503-8485.
`
`I hereby declare that all statements made herein of my own knowledge are true and that
`
`all statements made on information and belief are believed to be true; and further that these
`
`statements were made with the knowledge that willful false statements and the like so made
`
`are punishable by fine or imprisonment, or both, under Section 1001 of Title 18 of the United
`
`States Code and that such willful false statement may jeopardize the validity of the
`
`application or any patent issuing thereon.
`
`Sole inventor or
`first joint inventor:
`
`Full name
`
`Sylvain Cottens
`
`·o
`/ _,,... {)
`~~I·
`
`Signature
`Date
`Citizenship
`Residence
`
`Olat.e./ •. I~ I /j'J \
`Switzerland
`In den Reben 12, CH-4108 Witterswil,
`Switzerland
`~HX
`P.O. Address: same. as above ..
`
`IMPORTANT: Before this declaration is signed, the patent application (the specificateion, the claims and this
`declarations) must be read and understood by each person signing it, and no changes may be made in the
`application after this declaration has been signed.
`
`- 3 -
`
`
`
`

`
`Second joint inventor,
`if any:
`
`Full name
`
`100-7932/PCT
`
`Case No.
`oD
`Richard Sedrani ~ _,..,.,
`1Pl~4e-&\~
`
`a\.aMQ.. 11
`Luxembourg
`
`l ~ )_s-
`
`Signature
`Date
`Citizenship
`Residence
`
`Herrengrabenweg 15, CH-4054 Basle,
`6t/ X
`Switzerland
`P. 0. Address : same as above
`
`Third joint inventor,
`if any:
`
`Fourth joint inventor,
`if any:
`
`Fifth joint inventor
`if any:
`
`Full name
`
`Signature
`Date
`Citizenship
`Residence
`
`P.O. Address:
`
`Full name
`
`Signature
`Date
`Citizenship
`Residence
`
`P.O. Address:
`
`Full name
`
`Signature
`Date
`Citizenship
`Residence
`
`P.O. Address:
`
`- 4-
`
`
`
`

`
`!~
`•~ '
`
`.
`
`DO/EO BIBLIOGRAPHIC DATA ENTRY
`
`08 / 416673
`SERIAL N~~BER:
`IA NUMBER:
`PCT/ EP93 / 02604
`FAMILY NAME:
`COTTENS
`GIVEN NAME:
`SYLVAIN
`Y
`PRIORITY CLAIMED CY/NJ:
`NO BASIC FEE CY/NJ;
`N
`100-7932/PCT
`ATTORNEY DOCKET NUMBER:
`CORRESPONDENTS NAME/ADDRESS:
`ROBERT S. HONOR
`SANDOZ PATENT DEPT
`59 ROUTE 10
`E. HANOVER, N.J. 07936-1080
`
`04 / 07
`RECEIPT DATE:
`IA FILING DATE:
`09 / 24
`DELAY WAIVED (Y/NJ:
`N
`DEMAND RECEIVED (Y/NJ:
`Y
`PRIORITY DATE:
`10 / 09 /
`US DESIGNATED ONLY CY/NJ:
`COUNTRY:
`
`APPLICATION TITLES:
`0-ALKLATED RAPAMYCIN DERIVATIVES AND THEIR USE, PARTICULARLY AS
`IMMUNOSUPPRESSENTS
`
`OK TO UPDATE? <Y OR NJ Y
`
`N
`
`/ 95
`
`•
`•
`•
`/ q':( --.
`•
`t32 •
`£PX •
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`•
`
`
`
`

`
`.,i
`
`•~
`
`~B-AR_C_O-DE_LAB.....,E_L ____________ ....-;;~------,...---------------------~-~~--------------------------.
`
`I llllll llllll llll lllll lllll lllll lllll lllll llll llll
`
`U.S. PATENT APPLICATION
`
`SERIAL NUMBER
`
`RUNG DATE
`
`ClASS
`
`GROUP ART UNIT
`
`08/416,673
`
`04/07/95
`
`514
`
`1205
`
`SYLVAIN COTTENS, WITTERSWIL, SWITZERLAND; RICHARD SEDRANI, BASLE,
`SWITZERLAND.
`
`**CONTINUING DATA*********************
`VERIFIED
`THIS APPLN IS A 371 OF
`
`PCT/EP93/02604 09/24/93
`
`**FOREIGN/PCT APPLICATIONS************
`VERIFIED
`GREAT BRITAIN
`9221220.8
`
`10/09/92
`
`STATE DR
`COUNTRY
`
`SHEETS
`DRAWING
`
`TOTAL
`CLAIMS
`
`INDEPENDENT
`CLAIMS
`
`RUNG FEE
`RECEIVED
`
`ATTORNEY DOCKET NO.
`
`CHX
`
`0
`
`8
`
`1
`
`$1,090.00
`
`100-7932/PCT
`
`ROBERT S HONOR
`SANDOZ CORPORATION
`59 ROUTE 10
`EAST HANOVER NJ 07936-1080
`
`0-ALKYLATED RAPAMYCIN DERIVATIVES AND THEIR USE, PARTICULARLY AS
`IMMUNOSUPPRESSANTS
`
`This is to certify that annexed hereto is a true copy from the records of the United States
`Patent and Trademark Office of the application which is identified above.
`By authority of the
`COMMISSIONER OF PATENTS AND TRADEMARKS
`
`Date
`
`Certifying Officer
`
`
`
`

`
`PATENT APPLICATION SERIAL Qo~ I 4 l 6 6 7 a
`
`U.S. DEPARTMENT OF COMMERCE
`PATENT AND TRADEMARK OFFICE
`FEE RECORD SHEET
`
`----
`
`.::·-?~ •• ;:
`
`MS19049 04/12/95
`19039 05122195
`05/22/95
`19040
`
`08416673
`08416673
`08416673
`
`""s~1aoa6 05/30/95 08416673
`
`Hsi9046- 10iiJ195-Qa4i°66fa --
`
`PT0-1556
`(5/81)
`
`19-0134 190 960
`
`19--0134 190 960
`19-0134 190 970
`
`980.00CH
`980.00CR ~
`'
`980. OOCH V/(
`
`• 1-~--1-9~-~-03143-~ -~01~9-0 ~~906'"0--·---~~1~1-o-~--~.o~oc~\l /
`1 '50 ao ')'
`~
`
`•
`
`
`
`

`
`i,
`PATENT APPLICATION;~._ETERMINATION RECORD
`
`Effective 0 . ober 1, 1994
`
`-·
`
`l
`'pn~·:7r3;7fN
`
`CLAIMS AS FILED - PART I
`(Column 1)
`
`NUMBER FILED
`
`(Column 2)
`NUMBER EXTRA
`
`SMALL ENTITY
`
`OR
`
`RATE
`
`FEE
`
`OTHER THAN
`SMALL ENTITY
`
`RATE ~
`
`..
`
`FOR
`
`BASIC FEE
`
`TOTAL CLAIMS
`
`INDEPENDENT CLAIMS
`
`0
`
`~--
`
`S!ifJ / J minus 20 = *
`minus 3 =
`
`*
`
`p
`
`MULTIPLE DEPENDENT CLAIM PRESENT
`
`* If the difference in column 1 is less than zero, enter ·o· in column 2
`
`CLAIMS AS AMENDED - PART II
`(Column 1)
`(Column 2)
`CLAIMS
`REMAINING
`AFTER
`AMENDMENT
`
`HIGHEST
`NUMBER
`PREVIOUSLY
`PAID FOR
`
`~
`t-
`z
`w
`:E Total
`c
`z w Independent *
`:E
`~
`
`*
`
`Minus
`
`Minus
`
`**
`
`***
`
`y
`
`(
`
`(Column 3)
`
`PRESENT
`EXTRA
`
`.--;
`
`365.00
`
`x$11=
`
`x38=
`
`+120=
`
`TOTAL
`
`t.lU.UU
`
`OR
`OR x$22=
`
`I
`
`OR
`
`,x76=
`OR
`OR +240= b£¥V
`vc:.k
`TOTAL ru w ]
`OTH~~iA'i?
`
`SMALL ENTITY OR
`
`SMALL ENTITY
`
`RATE
`
`x$11=
`
`x38=
`
`ADDI-
`TIONAL
`FEE
`
`RATE
`
`ADDI-
`Tl ON AL
`FEE
`
`OR x$22=
`
`OR
`
`x76=
`
`=
`=
`FIRST PRESENTATION OF MULTIPLE DEPENDENT CLAIM
`
`(Column 1)
`
`(Column 2)
`
`(Column 3)
`
`"
`
`+120=
`TOTAL
`ADDIT. FEE
`
`OR +240=
`
`TOTAL
`OR ADDIT. FEE
`
`CLAIMS
`REMAINING
`AFTER
`AMENDMENT
`
`HIGHEST
`NUMBER
`PREVIOUSLY
`PAID FOR
`
`PRESENT
`EXTRA
`
`RATE
`
`ADDI-
`TIONAL
`FEE
`
`RATE
`
`ADDI-
`Tl ON AL
`FEE
`
`m
`t-
`z
`w
`:E Total
`c
`z
`w Independent *
`:E
`~
`
`*
`
`=
`=
`FIRST PRESENTATION OF MULTIPLE DEPENDENT CLAIM
`
`Minus
`
`Minus
`
`**
`
`***
`
`(Column 1)
`
`(Column 2)
`
`(Column 3)
`
`x$11=
`
`x38=
`
`+120=
`
`TOTAL
`ADDIT. FEE
`
`OR
`
`x$22=
`
`OR
`
`x76=
`
`OR +240=
`
`TOTAL
`OR ADDIT. FEE
`
`RATE
`
`ADDI-
`TIONAL
`FEE
`
`.
`ADDI-
`TIONAL
`FEE
`
`CLAIMS
`REMAINING
`AFTER
`AMENDMENT
`
`HIGHEST
`NUMBER
`PREVIOUSLY
`PAID FOR
`
`PRESENT
`EXTRA
`
`RATE
`
`0
`t-
`z
`w
`:E Total
`c
`z
`w Independent *
`:E
`~
`
`*
`
`=
`=
`FIRST PRESENTATION OF MULTIPLE DEPENDENT CLAIM
`
`Minus
`
`Minus
`
`**
`
`***
`
`x$11=-
`
`x38=
`
`+120=
`
`OR x$22=
`
`OR x76=
`
`OR +240=
`
`* If the entry in column 1 is less than the entry in column 2, write "O" in column 3.
`TOTAL
`TOTAL
`** If the "Highest Number Previously Paid For" IN THIS SPACE is less than 20, enter "20."
`OR ADDIT. FEE
`ADDIT. FEE
`***11 the Highest Number Previously Paid For" IN THIS SPACE is less than 3, enter "3."
`The Highest Number Previously Paid For" (Total or Independent) is the highest number found in the appropriate box in column 1.
`
`FORM PT0-875
`(Rev. 10/94)
`
`Patent and Trademark Office, U.S. DEPARTMENT OF COMMERCE
`
`
`
`

`
`W094/09010 •
`
`0-ALKYLAIEO RAPM!IY..C.ULD.ERIYATIV.ES_ANO_T.HEI.R_US.E., PARTICULARLY AS
`.
`. .
`'SUPPRESSANTS
`. ..
`·
`·
`/-~
`.
`::.i_,-· ~
`This invention comprises novel alkylated derivatives of rapamycin having
`\)
`
`-
`
`.-
`
`pharmaceutical utility. especially as immunosuppressants.
`
`Rapamycin is a known macrolide anti.biotic produced by Streptomvces
`
`hveToscooicus, having the structure depicted in Formula A:
`
`41
`
`5
`
`j
`
`0
`
`0---
`
`IMMUNO-
`
`(A)
`
`24
`
`See. e.g., McAlpine, J.B., et al., J. Antibiotics (1991) 44: 688; Schreiber, S.L., et al., J. Arn.
`
`Chem. Soc. (1991) 113: 7433; US Patent No. 3 929 992. Rapamycin is an extremely
`
`~.
`
`
`
`

`
`W094/09010
`
`PCT /EP93/02604
`
`- 2 -
`
`potent immunosuppressant and has also been shown to have antirumor and anrifungal
`
`activity. Its utility as a pharmaceutical. however, is restricted by its very low and variable
`
`bioavailability as well as its high toxicity. Moreover. rapamycin is highly insoluble, making
`
`it difficult to formulate stable galenic compositions.
`
`It has now surprisingly been discovered that cenain novel derivatives of rapamycin
`
`(the Novel Compounds) have an improved pharmacologic profile over rapamycin, exhibit
`
`greater stability and bioavailability, and allow for greater ease in producing galenic
`
`formulations. The Novel Compounds are alk:ylated derivatives of rapamycin having the
`
`srrucrure of Formula I:
`
`41
`
`50 ,, ... ··
`
`3
`
`: 34
`,,
`.:
`2
`6 7~0
`II
`N
`8
`
`0
`
`" ...
`
`0 , •.
`
`0---
`
`18
`
`20
`
`--
`
`wherein
`
`3
`
`(I)
`
`y
`
`24
`
`
`
`

`
`W094/09010
`
`PCT /EP93/02604
`
`-3-
`
`X is (RH) or O;
`
`Y is (H.OH) or O;
`
`R 1 and Rz are independently selected from
`. H. alkyl, thioalkyl, arylalkyl, hydroxyalkyl. dihydroxyalkyl,
`
`hydroxyalkylarylalkyl, dihydroxyalkylarylalkyl. alkoxyall..-yl, acyloxyalkyl.
`·---
`aminoalkyl, alkylaminoalkyl, a!.koxycarbonylaminoalkyl, acylaminoalkyl,
`
`arylsulfonamidoalkyl. allyl, dihydroxyalkylallyl, dioxola.nylallyl,
`) 3Si where each R3 is independently selected from H.
`carbalkoxyalk:yl, and (R3
`methyl, ethyl, isopropyl, 1-butyl, and phenyl; wherein "alk-" or "alkyl" refers
`to Ci.6 alkyl, branched or linear preferably C1•3 alkyl. in which the carbon
`chain may be optionally interrupted by an ether (-0-) linkage; and
`
`R' is methyl, or R~ and R1 together fomi ~ al.kylene;
`
`provided that R1 and R2 are not both H; and
`provided that where R1 is (R3
`) 3Si or carbalkoxyalky4 X and Y are not both 0.
`
`Preferred Novel Compounds include the following:
`40-0-Benzyl-rapamycin
`
`40-0-(4'-Hydroxymethyl)benzyl-rapamycin
`40-0-[ 4' -(1.2-Dillydroxyet:hyl)]benzyl-rapamycin
`
`40-0-Allyl-rapamycin
`
`40-0-[3'-(2,2-Dimethyl-l,3-dioxolan-4(S)-yl)-prop-2'-en-1'-yl]-rapamycin
`
`(2'E, 4'S)-40-0-(4' ,5'-Dihydroxypent-2'-en-l' -yl)-rapamycin
`
`40-0-(2-Hyd.roxy)ethoxycarbonylmerhyl-rapamycin
`
`40-0-(2-Hydroxy)ethyl-rapamycin
`
`1.
`
`2.
`3.
`
`4.
`
`S.
`6.
`
`7.
`
`&.
`
`
`
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`WO 94/09010
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`PCT/EP93/02604
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`- 4 -
`
`40-0-(3-Hydroxy)propyl-rapamycin
`
`40-0-(6-Hydroxy)hexyl-rapamycin
`
`40-0-[2-(2-Hydroxy)ethoxy]ethyl-rapamycin
`
`40-0-((35)-2,2-Dimethyldioxolan-3-yl]methyl-rapamycin
`
`40-0-((25)-2.3-Dihydroxyprop-l-yl]-rapamycin
`
`40-0-(2-Acetoxy )ethyl-rapamycin
`
`40-0-(2-Nicotinoyloxy)ethyl-rapamycin
`
`40-0-[2-(N-Morpholino )acetoxy ]ethyl-rapamycin
`
`; 40-0-(2-N-Imidazolylacetoxy)ethyl-rapamycin
`
`40-0-[2-(N-Methyl-N'-piperazinyl)acetoxy ]ethyl-rapamycin
`
`39-0-Desmethyl-39 ,40-0,0-ethylene-rapamycin
`
`(26R)-26-Dihydro-40-0-(2-hydroxy)ethyl-rapamycin
`
`28-0-Methyl-rapamycin
`
`40-0-(2-Aminoethyl)-rapamycin
`
`40-0-(2-Acetaminoethyl)-rapamycin
`
`40-0-(2-Nicotinamidoethyl)-rapamycin
`
`40-0-(2-(N-Methyl-imidazo-2'-ylcarbethoxamido)ethyl)-rapamycin
`
`40-0-(2-Ethoxycarbonylaminoethyl)-rapamycin
`
`40-0-(2-Tolylsulfonamidoethyl)-rapamycin
`
`40-0-[2-( 4' ,5 '-Dicarboethoxy-1 ',2 ',3 '-triazol-1 '-yl)-ethyl]-rapamycin
`
`9.
`
`10.
`
`11.
`
`12.
`
`13.
`
`14.
`
`15.
`
`16.
`
`17.
`
`18.
`
`19.
`
`20.
`
`21.
`
`22.
`
`23.
`
`24.
`
`25.
`
`26.
`
`27.
`
`28.
`
`The Novel Compounds for immunosuppressive use a.re preferably the
`40-0-substituted rapamycins where X and Y are both 0, R 2 is H. R4 is methyl, and R 1 is
`other than H; most preferably where R 1 is selected from hydroxyalkyl, hydroxyalkoxyalkyl,
`
`acylaminoalkyl, and aminoalkyl; especially 40-0-(2-hydroxy)ethyl-rapamycin, 40-0-(3-
`
`hydroxy)propyl-rapamycin, 40-0-[2-(2-hydroxy)ethoxy]ethyl-rapamycin, and
`
`40-0-(2-acetaminoethyl)-rapamycin).
`
`Preferably, 0-substitution at C40 or 0,0-disubstitution at C28 and C40 is performed
`
`
`
`

`
`W094/09010
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`PCT /EP93/02604
`
`- 5 -
`
`according to the following general process: Rapamycin (or dihydro or deoxorapamycin) is
`
`reacted with an organic radical attached to a leaving group (e.g., RX where R is the organic
`
`radical, e.g., an alkyl, allyl, or benzyl moiety, which is desired- as the 0-substiruent, and X is
`the leaving group, e.g., CC13C(NH)O or CF3S03) under suitable reaction conditions,
`preferably acidic or neutral conditions, e.g., in the presence of an acid like
`
`trifluoromethanesulfonic acid, camphorsulfonic acid, p-toluenesulfonic acid or their
`respective pyridinium or substituted pyridinium salts when X is CC13C(NH)O or in the
`presence of a base like pyridine, a substituted pyridine, diisopropylethylamine or
`pentamethylpiperidine when X is CF3S03•
`the same manner, but with prior protection at C40. Funher modifications are possible. For
`
`0-substirutions at C28 only are accomplished in
`
`example, where the substiruent is allyl, the isolated, monosubstituted double bond of the
`
`allyl moiety is highly amenable to funher modification.
`
`The 9-deoxorapamycin compounds are preferably produced by reducing a rapamycin
`
`using hydrogen sulfide, by reacting rapamycin with diphenyldiselenide and tributylphosphine
`
`or by other suitable reduction reaction.
`
`The 26-dihydro-rapamycins are preferably produced by reducing rapamycins or
`
`9-deoxorapamycins from keto to hydroxy at C26 by a mild reduction reaction, such as a
`
`borohydride reduction reaction.
`
`The Novel Compounds are particularly useful for the following conditions:
`
`a)
`
`Treatment and prevention of organ or tissue transplant rejection, e.g. for the
`
`treatment of recipients of e.g. heart, lung, combined heart-lung, liver, kidney, pancreatic,
`
`skin or corneal transplants. They are also indicated for the prevention of graft-versus-host
`
`disease, such as following bone marrow transplantation.
`
`b)
`
`Treatment and prevention of autoimmune disease and of inflammatory
`
`conditions, in particular inflammatory conditions with an etiology including an autoimmune
`
`
`
`

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`WO 94/09010
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`••
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`
`- 6 -
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`component such as anhritis (for example rheumatoid anhritis, anhritis chronica progrediente
`
`and anhritis deformans) and rheumatic diseases. Specific autoimmune diseases for which
`
`the compounds of the invention may be employed include, autoimmune hematological
`
`disorders (including e.g. hemolytic anaemia, aplastic anaemia. pure red cell anaemia and
`
`idiopathic thrombocytopenia), systemic lupus erythematosus, polychondritis, sclerodoma,
`
`Wegener granulamatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis,
`
`psoriasis, Steven-Johnson syndrome, idiopathic sprue, autoimmune inflammatory bowel
`disease (including e.g. ulcerative colitis and Crohn's disease) endocrine ophthalmopathy,
`
`Graves disease, sarcoidosis, multiple sclerosis, primary billiary cirrhosis, juvenile diabetes
`
`(diabetes mellitus type I), uveitis (anterior and posterior), keratoconjunctivitis sicca and
`
`vernal keratoconjunctivitis, interstitial lung fibrosis, psoriatic anhritis, glomerulonephritis
`
`(with and without nephrotic syndrome, e.g. including idiopathic nephrotic syndrome or
`
`minimal change nephropathy) and juvenile dermatomyositis.
`
`c)
`
`d)
`
`Treatment and prevention of asthma.
`
`Treatment of multi-drug resistance (MDR). The Novel Compounds suppress
`
`P-glycoproteins (Pgp), which are the membrane transpon molecules associated with MDR.
`
`MDR is particularly problematic in cancer patients and AIDS patients who will not respond
`
`to conventional chemotherapy because the medication is pumped out of the cells by Pgp.
`
`The Novel Compounds are therefore useful for enhancing the efficacy of other
`
`chemotherapeutic agents in the treatment and control of multi.drug resistant conditions such
`
`as multi.drug resistant cancer or multi.drug resistant AIDS.
`
`e)
`
`Treatment of proliferative disorders, e.g. tumors, hyperproliferative skin
`
`disorder and the like.
`
`f)
`
`g)
`
`of steroids.
`
`Treatment of fungal infections.
`
`Treatment and prevention of inflammation, especially in potentiating the action
`
`h)
`
`Treatment and prevention of infection, especially infection by pathogens
`
`having Mip or Mip-like factors.
`
`i)
`
`Treatment of overdoses of FK-506, rapamycin, immunosuppressive Novel
`
`7
`
`
`
`

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`W094/09010
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`-
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`- 7 -
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`PCT /EP93/02604
`
`Compounds, and other macrophilin binding immunosuppressants.
`
`The invention thus provides the Novel Compounds described herein, for use as novel
`
`intermediates or as pharmaceuticals, methods of treating or preventing the above-described
`
`disorders by administering an effective amount of a Novel Compound to a patient in need
`
`thereof, use of a Novel Compound in the manufacture of a medicament for treaonent or
`
`prevention of the above-described disorders, and pharmaceutical compositions comprising a
`
`Novel Compound in combination or association with a pharmaceutically acceptable diluent
`
`or carrier.
`
`Most of the Novel Compounds described herein are highly immunosuppressive,
`
`especially those Novel Compounds which are 0-substituted at C40, and these Novel
`
`Compounds are panicularly useful in indications a and b, but not in indication i. Those of
`
`the Novel Compounds which are less immunosuppressive, especially those which are 0-
`
`substituted at C28 only, are particularly useful in indications h and i, but are less preferred
`
`in indications a orb.
`
`The Novel Compounds are utilized by administration of a pharmaceutically effective
`
`dose in pharmaceutically acceptable fonn to a subject in need of treattnent. Appropriate
`
`dosages of the Novel Compounds will of course vary, e.g. depending on the condition to be
`
`treated (for example the disease type or the nature of resistance), the effect desired and the
`
`mode of administration.
`
`In general however satisfactory results are obtained on administration orally at
`
`dosages on the order of from 0.05 to 5 or up to 1 Omg/kglday, e.g. on the order of from 0.1
`
`to 2 or up to 7.5 mg/kg/day administered once or. in divided doses 2 to 4x per day, or on
`
`administration parenterally. e.g. intravenously, for example by i.v. drip or infusion, at
`
`dosages on the order of from 0.01 to 2.5 up to 5 mg/kg/day. e.g. on the order of from 0.05
`
`or 0.1 up to 1.0 mg/kg/day. Suitable daily dosages for patients are thus on the order of 500
`
`t
`
`
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`

`
`W094/09010 •
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`PCT /EP93/02604
`
`- 8 -
`
`mg p.o., e.g. on the order of from 5 to 100 mg p.o., or on the order of from 0.5 to 125 up to
`
`250 mg i.v~. e.g. on the order of from 2.5 to 50 mg i.v ..
`
`Alternatively and even preferably, dosaging is arranged in patient specific manner to
`
`provide pre-determined trough blood levels, e.g. as determined by RIA technique. Thus
`
`patient dosaging may be adjusted so as to achieve regular on-going trough blood levels as
`
`measured by RIA on the order of from 50 or 150 up to 500 or lOOOng/ml, i.e. analogously to
`
`methods of dosaging currently employed for Ciclosporin immunosuppressive therapy.
`
`The Novel Compounds may be administered as the sole active ingredient or together
`
`with other drugs. For example, in immunosuppressive applications such as prevention and
`
`treatment of graft vs. host disease, transplant rejection, or autoimmune disease, the Novel
`
`Compounds may be used in combination with Ciclosporin, FK-506, or their
`
`immunosuppressive derivatives; corticosteroids; azathioprene; immunosuppressive
`
`monoclonal antibodies, e.g., monoclonal antibodies to CD3, CD4, CD25, CD28, or CD45;
`
`and7or other immunomodulatory compounds. For anti-inflammatory applications, the Novel
`
`Compounds can be used together with anti-inflammatory agents, e.g., corticosteroids. For
`
`anti-infective applications, the Novel Compounds can be used in combination with other
`
`anti-infective agents, e.g., anti-viral drugs or antibiotics.
`
`The Novel Compounds are administered by any conventional route, in particular
`
`enterally, e.g. orally, for example in the form of solutions for drinking, tablets or capsules or
`
`parenterally, for example in the form of injectable solutions or suspensions. Suitable unit
`
`dosage forms for oral administration comprise, e.g. from 1 to 50 mg of a compound of the
`invention, usually 1 to 10 mg. Pharmaceutical compositions comprising the novel
`
`compounds may be prepared analogously to pharmaceutical compositions comprising
`
`rapamycin, e.g., as described in EPA 0 041 795, which would be evident to one skilled in
`the an.
`
`9
`
`
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`

`
`. WO 94/09010
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`••
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`PCT /EP93/02604
`
`- 9 -
`
`The phannacological activity of the Novel Compounds are demonstrated in, e.g., the
`
`following tests:
`
`1. Mixed lvmphocvte reaction CMLR)
`
`The Mixed Lymphocyte Reaction was originally developed in connection with
`
`allografts, to assess the tissue compatibility between potential organ donors and recipients,
`
`and is one of the best established models of immune reaction in vitro. A murine model
`
`MLR, e.g., as described by T.Meo in "Immunological Methods", L. Lefkovits and B. Peris,
`
`Eds., Academic Press, N.Y. pp. 227-239 (1979), is used to demonstrate the
`immunosuppressive effect of the Novel Compounds. Spleen cells (0.5 x 106
`) from Balb/c
`mice (female, 8-10 weeks) are co-incubated for 5 days with 0.5 x 106 irradiated (2000 rads)
`
`or mitomycin C treated spleen cells from CBA mice (female, 8-10 weeks). The irradiated
`
`allogeneic cells induce a proliferative response in the Balb/c spleen cells which can be
`
`measured by labeled precursor incorporation into the DNA. Since the stimulator cells are
`
`irradiated (or mitomycin C treated) they do not respond to the Balb/c cells with proliferation
`
`but do retain their antigenicity. The antiproliferative effect of the Novel Compounds on the
`
`Balb/c cells is measured at various dilutions and the concentration resulting in 50%
`inhibition of cell proliferation (IC50) is calculated. The inhibitory capacity of the test sample
`may be compared to rapamycin and expressed as a relative IC50 (i.e. ICso test sample/IC50
`rapamycin).
`
`2.
`
`IL-6 mediated proliferation
`
`The capacity of the Novel Compounds to interfere with growth factor associated
`
`signalling pathways is assessed using an interleukin-6 (IL-6)-dependent mouse hybridoma
`
`cell line. The assay is performed in 96-well microtiter plates. 5000 cells/well are cultivated
`
`in serum-free medium (as described by M. H. Schreier and R. Tees in Immunological
`
`Methods, I. Lefkovits and B. Pemis, eds., Academic Press 1981, Vol. II, pp. 263-275),
`
`supplemented with 1 ng recombinant IL-6/ml. Following a 66 hour incubation in the
`
`absence or presence of a test sample, cells are pulsed with 1 µCi (3-H)-thymid.ine/well for
`
`/O.
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`PCT /EP93/02604
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`- IO -
`
`another 6 hours, harvested and counted by liquid scintillation. (3-H)-thymidine incorporation
`
`into DNA correlates with the increase in cell number and is thus a measure of cell
`
`proliferation. A dilution series of the test sample allows the calculation of the concentration
`resulting in 50% inhibition of cell proliferation (IC50). The inhibitory capacity of the test
`sample may be compared to rapamycin and expressed as a relative IC50 (i.e. IC50 test
`sam