Illlllllllllllllllllllllllllllllllllll
`
`US008129342B2
`US008129342B2
`
`(12) United States Patent
`(12) United States Patent
`Kelleher et al.
`Kelleher et a].
`
`(io) Patent No.:
`(16) Patent No.:
`(45) Date of Patent:
`(45) Date of Patent:
`
`US 8,129,342 B2
`US 8,129,342 B2
`*Mar. 6, 2012
`*Mar. 6, 2012
`
`6/1999 Baker
`5,912,226 A
`5,912,226 A
`6/1999 Baker
`9/1999 Webber
`5,955,509 A
`5,955,509 A
`9/1999 Webber
`2/2001 Hammann
`6,194,383 B1
`3%? et 31
`10/2002 Oleson, Jr. et al.
`6,468,967 B1
`2/2004 Kelleher
`6,696,412 B1
`2/2004 Kelleher
`6,696,412 B1
`2/2005 Oleson, Jr. et al.
`6,852,689 B2
`2/2005 Oleson, Jr. et al.
`6,852,689 B2
`4/2006 Baker et al
`514/2.3
`RE39,071 E *
`
`
`4/2006 Baker et 31' RE39’071 E : 8,058,238 B2 11/2011 Kelleher et al. ............. .. 435/886
`11/2011 Kelleher et al. ..
`435/886
`8,058,238 B2 *
`FOREIGN PATENT DOCUMENTS
`FOREIGN PATENT DOCUMENTS
`095295 Al
`11/1983
`095295 A1
`11/1983
`178152 A2
`4/1986
`178152 A2
`4/1986
`294990 A2
`12/1988
`10/1989
`337731 B1
`9/1990
`386951 A2
`9/1990
`386951 A2
`WO 99/27954
`6/1999
`6/1999
`WO 99/27954
`6/1999
`WO 99/27957
`WO 99/27957
`6/1999
`WO 99/40113
`8/1999
`8/1999
`W0 99/40ll3
`WO 99/43700
`9/1999
`9 1999
`W0 99 43700
`4/2000
`WO 00/18419
`4/2000
`WO 00/18419
`WO 00/81419
`4/2000
`4/2000
`W0 00/81419
`WO 01/44271
`6/2001
`6/2001
`WO 01/44271
`WO 01/44272
`6/2001
`6/2001
`WO 01/44272
`WO 01/44274
`6/2001
`6/2001
`WO 01/44274
`WO 01/53330
`7/2001
`
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`EP
`WO
`W0
`W0
`WO
`WO
`W0
`WO
`W0
`W0
`WO
`WO
`W0
`WO
`W0
`WO
`W0
`WO
`W0
`WO
`
`OTHER PUBLICATIONS
`OTHER PUBLICATIONS
`
`g
`y
`Agreement between Cubist Pharmaceuticals, Inc. and Eli Lilly and
`A reement between Cubist Pharmaceuticals, Inc. and Eli Lill and
`Company dated Nov. 7, 1997 (redacted form from SEC Edgar).
`Company dated NOV. 7, 1997 (redacted form from SEC Edgar).
`Agreement between Cubist Pharmaceuticals, Inc. and Eli Lilly and
`Agreement between Cubist Pharmaceuticals, Inc. and Eli Lilly and
`Company dated Oct. 6, 2000 (redacted form from SEC Edgar).
`Company dated Oct. 6, 2000 (redacted form from SEC Edgar).
`Assignment of US RE 39,071 from Eli Lilly and Company to Cubist
`Assignment of US RE 39,071 from Eli Lilly and Company to Cubist
`Pharmaceuticals, Inc. recorded on Apr. 23, 2007 Reel/Frame:
`Pharmaceuticals, Inc. recorded on Apr. 23, 2007 Reel/Frame:
`019l81/0916‘
`019181/0916.
`Debono, M‘ et a1‘; “Enzymatic and Chemical Modi?cations of
`Debono, M. et al.; "Enzymatic and Chemical Modifications of
`Lipopeptide Antibiotic A21978C: The Synthesis and Evaluation of
`Lipopeptide Antibiotic A21978C: The Synthesis and Evaluation of
`Daptomycin (LY146032)," J. Antibiotics; 41; 1988; pp. 1093-1105.
`Daptomycin (LY146032),” J. Antibiotics; 41; 1988; pp. 1093-1105.
`_
`(Continued)
`(Continued)
`
`Primary Examiner — Chih-Min Kam
`Primary Examiner * Chih-Min Kam
`(74) Attorney, Agent, or Firm — Cubist Pharmaceuticals,
`(74) Attorney, Agent, or Firm *Cubist Pharmaceuticals,
`Inc.
`1110
`
`(54) HIGH PURITY LIPOPEPTIDES
`(54) HIGH PURITY LIPOPEPTIDES
`
`Inventors: Thomas J. Kelleher, Thousand Oaks,
`(75)
`(75) Inventors: Thomas J. Kelleher, Thousand Oaks,
`CA (US); Jan-Ji Lai, Westborough, MA
`CA (Us); JaII-Ji Lai, Westborough, MA
`(US); Joseph P. DeCourcey, Boston,
`(US); Joseph P. DeCourcey, Boston,
`MA (US); Paul D. Lynch, Arlington,
`
`MA (US); Paul D. Lynch, Arlington, MA (Us); Maurizio Zenoni’ Ferentino
`MA (US); Maurizio Zenoni, Ferentino
`Frosinone (IT); Auro R. Tagliani, Pavia
`Frosinone (IT); Auro R. Tagliani, Pavia
`(IT)
`(1T)
`
`( * ) Notice:
`( * ) Notice:
`
`(73) Assignee: Cubist Pharmaceuticals, Inc.,
`
`(73) Assignee: Cubist Pharmaceuticals, Inc., Lexington MA (US)
`Lexington, MA (US)
`’
`Subject to any disclaimer, the term of this
`Subject to any disclaimer, the term of this
`patent is extended or adjusted under 35
`patent is extended or adjusted under 35
`U.S.C. 154(b) by 0 days.
`U'S'C' 154(1)) by 0 days‘
`This patent is subject to a terminal dis­
`This patent is subject to a terminal dis-
`claimer.
`Claimer
`'
`(21) Appl.No.: 12/888,233
`
`(22) Filed:
`(22) F1led:
`(65)
`(65)
`
`Sep. 22, 2010
`Sep. 22, 2010
`Prior Publication Data
`Prior Publication Data
`I
`I
`I
`US 2011/0207658 Al
`Aug. 25, 2011
`Us 201 1/0207658 A1
`Aug' 25’ 2011
`
`Related U.S. Application Data
`Related U_s_ Application Data
`_
`_
`_
`_
`(63) Continuation of application No. 11/739,180, filed on
`(63) cont1nuat1on of apphcanon NO‘ 11/739’180’ _?1ed_ On
`Apr. 24, 2007, now Pat. No. 8,058,238, which is a
`Apr.‘ 24, ‘2007, noW'Pat'. No. 8,058,238, Wh1ch 1s a
`continuation of application No. 10/747,485, filed on
`cont1nuat1on of application No. 10/747,485, ?led on
`Dec. 29, 2003, now abandoned, which
`is a
`Dec: 293 2003, 110W‘ abandoned, Whlch 15 a
`continuation of application No. 09/735,191, filed on
`cont1nuat1on 0f apphcatlon NO- 09/735,191, ?led 0n
`Nov. 28, 2000, now Pat. No. 6,696,412.
`Nov. 28, 2000, noW Pat. No. 6,696,412.
`(60) Provisional application No. 60/177,170, filed on Jan.
`(60) Provisional application No. 60/177,170, ?led on Jan.
`20, 2000.
`20, 2000.
`
`(51) Int. CI.
`(51) Int- Cl-
`(2006.01)
`€07K 7/50
`(2006.01)
`C07K 7/50
`(2006.01)
`C07K 7/00
`(2006.01)
`C07K 7/00
`(52) U.S. CI
`514/9; 514/11; 514/2; 514/14;
`(52) U.S. Cl. .................. .. 514/9; 514/11; 514/2; 514/14;
`530/317; 530/322; 530/344; 435/886
`530/317; 530/322; 530/344; 435/886
`(58) Field of Classification Search
`514/9, 11,
`(58) Field of Classi?cation Search .............. .. 514/9, 11,
`514/2’ 14; 530/317’ 322’ 344; 435/886
`514/2, 14; 530/317, 322, 344; 435/886
`See application ?le for complete search history.
`See application file for complete search history.
`
`(56)
`(56)
`
`References Cited
`References Cited
`
`U.S. PATENT DOCUMENTS
`U.S. PATENT DOCUMENTS
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`5/1997 Lattrell
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`.
`
`.
`
`.
`
`ABSTRACT
`(57)
`ABSTRACT
`(57)
`.
`.
`.
`The invention discloses highly purified daptomycin and to
`The 1nvent1on d1scloses ~highly puri?ed daptomycin and to
`pharmaceutical compositions comprising this compound.
`phanpaceullcal .composmons compnsmg lhls compounq'
`The invention discloses a method of purifying daptomycin
`The invention d1scloses a method of purifying daptomyc1n
`comprising the sequential steps of anion exchange chroma­
`comprising the sequential steps of anion exchange chroma
`tography, hydrophobic interaction chromatography and
`tography, hydrophobic interaction chromatography and
`anion exchange chromatography. The invention also dis­
`anion exchange chromatography. The invention also dis
`closes a method of purifying daptomycin by modified buffer
`closes a method of purifying daptomycin by modi?ed buffer
`enhanced anion exchange chromatography. The invention
`enhanced anion exchange chromatography. The invention
`also discloses an improved method for producing daptomycin
`also discloses an improved method for producing daptomycin
`by fermentation of Streptomyces roseosporus. The invention
`by fermentation of Streptomyces roseosporus. The invention
`also discloses high pressure liquid chromatography methods
`also discloses high pressure liquid chromatography methods
`for analysis of daptomycin purify. The invention also dis­
`for analysis of daptomycin purity. The invention also dis
`closes lipopeptide micelles and methods of making the
`closes lipopeptide micelles and methods of making the
`micelles. The invention also discloses methods of using
`micelles. The invention also discloses methods of using
`lipopeptide micelles for purifying lipopeptide antibiotics,
`lipopeptide micelles for purifying lipopeptide antibiotics,
`such as daptomycin. The invention also discloses using
`such as daptomycin. The invention also discloses using
`lipopeptide micelles therapeutically.
`lipopeptide micelles therapeutically.
`
`54 Claims, 11 Drawing Sheets
`54 Claims, 11 Drawing Sheets
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 1 of 34
`
`

`

`US 8,129,342 B2
`US 8,129,342 B2
`Page 2
`Page 2
`
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`Cubicin® (daptomycin for injection) Label 1004—Sep. 2003.
`Cubicin® (daptomycin for injection) Label 1004*Sep. 2003.
`Cubicin® (daptomycin for injection) Label 1004-1—Revised Aug.
`Cubicin® (daptomycin for injection) Label 1004-1iRevised Aug.
`2004.
`2004.
`Cubicin® (daptomycin for injection) Label 1004-2—Revised Jun.
`Cubicin® (daptomycin for injection) Label 1004-2iRevised Jun.
`2005.
`2005.
`Cubicin® (daptomycin for injection) Label 1004-10-1 Aug. 2010.
`Cubicin® (daptomycin for injection) Label 1004-10-1 Aug. 2010.
`• cited by examiner
`* cited by examiner
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 2 of 34
`
`

`

`U.S. Patent
`
`Mar. 6, 2012
`
`Sheet 1 of 11
`
`US 8,129,342 B2
`
`O
`
`M
`
`NHo
`
`I ^
`
`HO2C
`
`HN
`
`NH
`
`o o
`
`O
`
`0.
`
`HO
`
`NH
`
`O
`
`H02C
`
`HN
`
`O
`
`O
`
`HN
`
`N
`H
`0 H02C
`
`O
`
`H
`
`NH
`
`O
`
`HN
`H
`N
`
`O
`
`(CH2)8CH3
`
`CON Hp
`O
`
`II
`o
`'C02H
`
`H
`
`O
`
`RJ
`
`'/
`| |
`
`NH2
`
`Fig. 1
`
`O
`ll
`
`NH,
`l ^
`
`HO2C
`
`HN
`
`HO
`
`NH
`
`O O
`
`O
`
`NH
`
`O
`
`NH OH
`O
`
`O
`
`0
`
`HN
`
`N
`H
`0 HO2C
`
`O.
`
`O
`
`O
`
`N
`H
`
`NH
`
`0
`
`HN
`
`H
`N
`
`O
`
`H
`N
`
`O
`'C02H
`
`NH2
`
`Fig. 2
`
`(CHg^CHg
`
`.CONHo
`O
`
`N
`H
`
`O
`N —^
`
`'/
`N
`H
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 3 of 34
`
`

`

`U.S. Patent
`
`Mar. 6, 2012
`
`Sheet 2 of 11
`
`US 8,129,342 B2
`
`O
`
`NHp XA
`
`HO2C
`HN
`
`NH
`
`HO
`
`O O
`
`O
`
`NH
`
`O
`
`N
`
`O
`
`O
`
`O
`
`HN
`
`N
`H
`O H02C
`
`(CHgJgCHg
`
`H
`N
`fl
`
`O
`
`N
`H
`
`CONHp
`O
`
`N
`H
`
`O.
`
`O
`
`N
`H
`
`NH
`
`O
`
`HN
`H
`N
`
`0
`
`O
`
`H
`N
`
`O
`'CO2H
`
`NH2
`
`Fig. 3
`
`O
`
`I I
`
`NHo
`
`i ^
`
`NH
`
`OH
`O HO
`
`O
`
`O
`
`N
`H
`
`NH
`
`O
`
`HN
`H
`N
`
`HO2C
`
`HN
`
`HO
`
`O O
`
`NH
`
`O
`
`H
`
`HO2C
`2\
`
`o
`
`O
`
`HN
`
`N
`H
`O H02C
`
`,CONH2 o
`N
`H
`
`O
`
`H
`N
`
`(C^JaCHa
`
`N
`H
`
`H
`N
`
`O
`'C02H
`
`NH2
`
`Fig. 4
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 4 of 34
`
`

`

`U.S. Patent
`US. Patent
`
`Mar. 6, 2012
`Mar. 6, 2012
`
`Sheet 3 of 11
`Sheet 3 or 11
`
`US 8,129,342 B2
`US 8,129,342 B2
`
`O
`
`I
`
`I
`
`NHo
`NH2
`
`^
`
`i
`
`
`
`OH
`OH
`O HO
`0
`O H
`O
`O
`
`n
`N
`H
`
`O H
`O
`H
`N
`N
`O
`O
`'C02H
`co2H
`
`NH
`NH
`
`HO2C
`H020
`HN
`HN
`HO
`O O
`Hojo O
`NH
`NH
`
`O
`O
`
`HNOH
`O
`
`0
`
`o
`
`HN
`
`N
`H
`0 H02C
`
`NH
`NH
`O
`
`3:0
`
`HN
`HN
`
`H
`N
`
`O
`
`NH2
`NH2
`
`Fig. 5
`
`CONH2
`CONHo
`O
`O
`0 Hi
`H
`(CH2)8CH3
`N
`N
`(CH2)8CH3
`N
`N
`H
`
`H % N
`
`N
`H
`H
`
`HO
`O
`H G
`O
`
`{TEN/yr
`
`O
`
`HN w Lk
`
`H
`N
`
`O
`CO2H
`0 H2
`
`(CH2)8CH3
`
`CONHo
`0
`
`NH
`N
`H
`
`O
`
`H
`N
`
`'/
`N
`H
`
`N
`H
`NH
`NH O H
`O
`
`HO
`
`Fig. 6
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 5 of 34
`
`

`

`U.S. Patent
`
`Mar. 6, 2012
`
`Sheet 4 of 11
`
`US 8,129,342 B2
`
`0
`
`NH2
`
`HO2C
`HN
`
`HO
`
`NH
`
`o o
`
`O
`
`NH
`
`O
`
`HN
`
`HOoC
`2 \
`
`HN
`
`O
`N/ o
`
`O
`
`I—N
`H
`NH
`
`H
`N
`
`O
`'C02H
`
`CONHp
`0
`
`N
`H
`
`(CH2)QCH3
`
`O
`H U
`N
`
`0
`H2N
`
`O
`
`O
`
`0 HO2C^
`
`O
`
`HN
`H
`N
`
`O
`
`NH2
`
`Fig. 7
`
`0
`
`Hi
`
`H
`N
`
`O
`'C02H
`
`CONHo
`O
`
`N
`H
`
`O
`H U
`N
`
`(CH2)8CH3
`
`N
`H
`
`Fig. 8
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 6 of 34
`
`

`

`U.S. Patent
`
`Mar. 6, 2012
`
`Sheet 5 of 11
`
`US 8,129,342 B2
`
`o
`
`NH2
`
`V
`
`O
`
`o
`
`HO2C
`
`NH
`
`HN
`
`O O
`
`NH
`
`HO
`\
`
`O
`
`HN
`
`HOoC
`2 \
`
`HN
`
`O
`
`O
`
`O
`
`N
`H
`H02C
`
`CON Ho
`O
`
`N
`H
`
`O
`
`H
`N
`
`(CH2)7CH3
`
`N
`H
`
`0
`
`H
`N
`
`O
`•C02H
`
`NH2
`
`O
`
`N
`H
`NH
`
`O
`
`HN
`H
`N
`
`O
`
`Fig. 9
`
`HO
`
`O
`
`=c
`
`>—N
`H
`NH
`
`O
`
`HN
`
`HO
`
`O
`
`CONH?
`r o
`N
`H
`
`o
`a ji
`N
`
`(CH2)8CH3
`
`o
`
`H
`
`N n
`
`O
`'C02H
`
`N
`H
`
`NH2
`
`Fig. 10
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 7 of 34
`
`

`

`U.S. Patent
`
`Mar. 6, 2012
`
`Sheet 6 of 11
`
`US 8,129,342 B2
`
`O
`
`I
`
`I
`
`
`
`NHo
`
`I
`
` ^
`
`CON Hp
`0 , - 0
`N-MCH^CHCHgJCHg
`
`O
`
`A/ H
`N
`N
`Y - N
`H
`H
`O
`'C02H
`NH
`
`'/
`N
`H
`
`HO2C
`
`NH
`
`HN
`
`o o
`
`NH
`
`O.
`
`O
`
`O
`
`HO
`\
`
`O
`
`HN
`
`HOoC
`2 \
`
`HN
`
`O
`
`O
`
`N
`H
`O HO2C-
`
`O
`
`HN
`H
`N
`
`O
`
`NH2
`
`Fig. 11
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 8 of 34
`
`

`

`00
`CZ5
`d
`
`so
`K>
`s*
`
`US 8,129,342 B2
`
`K> w K>
`
`4^
`
`o
`<1
`re
`re
`cr
`5/3
`
`Sheet 7 or 11
`
`K>
`o
`K>
`o\
`S
`
`Mar. 6, 2012
`
`p
`
`C/5
`
`S3
`n
`P
`
`US. Patent
`
`010ml
`
`-29.40
`
`-24.53
`-23.11
`
`-20.93
`-19.57
`-17.92
`
`-15.10
`-14.43
`-13.10
`-12.28
`-11.54
`
`Z3-16.68
`
`-9.11
`-7.96
`-7.34
`-6.10
`
`i
`
`(71
`O
`
`1
`o
`o
`
`= 10/11
`
`1 :BF
`
`10 O =:
`
`-=12
`
`1 NF
`
`OSJ9
`
`= 6/7
`
`Fo -34
`3
`
`3
`m
`CQ 5
`31 g
`
`ro
`
`oo -=1
`
`111111
`Ol
`<_n
`
`1.:
`
`i
`O
`Cn
`
`-+p
`>-P
`
`Q\
`
`6
`
`ro -=
`
`Cn
`
`% w 5 5 4 4 3 3 2 2 l }
`
`O
`
`mhi
`
`In
`CO
`
`O
`CO
`
`5 O 5 0 5 O 5 O 5 O 5 0
`
`on
`ND
`
`O
`ND
`
`111111
`
`11
`
` M111
`
`i
`
`Cn
`
`o
`
`Cn
`
`o
`
`RESPONSE (mV)
`
`81012141618 20 22 24 26 28
`TlME(min)
`Fig. 12
`
`03 —
`Kl
`
`-=13
`
`*• 314
`K)
`
`1 Q
`
`| S.
`
`N3 ~B
`K)
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 9 of 34
`
`

`

`sO
`K>
`
`00
`C/2
`d
`
`US 8,129,342 B2
`
`w K>
`
`K>
`•1^
`
`Sheet 8 or 11
`
`o
`00
`2-
`«
`ST
`5/3
`
`s
`
`K>
`K> O
`o\
`P
`
`Mar. 6, 2012
`
`C/5
`
`ss
`n
`p
`
`US. Patent
`
`Fig. 13
`
`9 .mE
`
`28
`
`mu
`
`26
`
`0N
`
`24
`
`vm
`
`22
`
`mu
`
`T
`
`20
`
`ow
`
`T
`
`i—r
`
`T
`
`r
`
`T
`
`1
`+
`a. 2 S
`
`-10-=
`
`0-E
`
`10-^
`
`ns
`UJ
`co
`a.
`z O 30-=
`to
`LU
`E 40^
`>
`50^
`
`20 —
`
`ON
`
`on
`
`om
`
`(AUJ) EISNOdSEIEI
`
`CM
`r-^
`<0
`o
`
`CM
`•t
`p*1"^
`r^
`
`S
`o
`
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`
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`
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`
`60^
`
`on
`
`70-S
`
`on
`
`80-=
`
`cm
`
`90-^
`
`00
`
`TIME (min)
`18
`T
`
`
`
`E2: 52;
`
`2
`
`16
`
`14
`
`12
`
`10
`
`8
`IT
`
`6
`
`4
`
`2 3
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 10 of 34
`
`

`

`U.S. Patent
`US. Patent
`
`Mar. 6, 2012
`Mar. 6, 2012
`
`Sheet 9 of 11
`Sheet 9 or 11
`
`US 8,129,342 B2
`US 8,129,342 B2
`
`© ©«©, ? ©
`©
`©
`©
`
`©
`^ ©
`©
`©
`Fig. 14A
`
`teoofli
`o
`Fig. 14B
`
`o
`
`o
`
`Oc^O o AQUEOUS Qr3
`CtpD
`
`Fig. 14C
`Fig. 14C
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 11 of 34
`
`

`

`U.S. Patent
`US. Patent
`
`Mar. 6, 2012
`Mar. 6, 2012
`
`Sheet 10 of 11
`Sheet 10 or 11
`
`US 8,129,342 B2
`US 8,129,342 B2
`
`10CH
`
`
`
`EuG: .>.:>CUDOZOU
`
`E 80-
`u
`i/)
`D
`- 60-
`>
`a 40-
`D Q
`O 20-
`U
`
`(—
`
`0 "
`
`CMC
`
`I
`f
`:
`
`MIN":
`
`T
`T
`T
`T
`I
`I
`I
`0
`8000
`6000
`4000
`2000
`10000 12000
`8000
`6000
`4000
`2000
`10000 12000
`0
`DAPTOMYCIN CONCENTRATIOK ug/mL
`DAPTOMYCIN CONCENTRATION, ug/mL
`Fig. 15
`Fig. 15
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 12 of 34
`
`

`

`U.S. Patent
`US. Patent
`
`Mar. 6, 2012
`Mar. 6, 2012
`
`Sheet 11 of 11
`Sheet 11 0f 11
`
`US 8,129,342 B2
`US 8,129,342 B2
`
`INTENSITY
`INTENSITY
`SIZE DISTRIBUTION(S)
`SIZE DISTRIBUTIONS]
`
`40-
`
`m $
`
`U
`2
`SS 20-
`
`KI
`
`l;.
`I'"''I
`
`: :.V
`
`fo
`5 10
`5
`
`•"t
`B.
`.==+
`5'0 100
`50 100
`500
`500
`DIAMETER (nm)
`DIAMETER (nm)
`
`VOLUME
`VOLUME
`
`SIZE DISTRIBUTIONS)
`SIZE DISTRIBUTIONlS)
`
`40-
`
`to
`
`U
`z
`
`v - t
`
`' • 3 . ^
`
`5 10
`
`I
`
`I
`
`50 100
`500
`560 '
`50 100
`DIAMETER {nm)
`DIAMETER (nm)
`V
`Fig. 16
`Fig. 16
`
`J
`
`PETITIONERS
`
`EXHIBIT NO. 1002 Page 13 of 34
`
`

`

`US 8,129,342 B2
`US 8,129,342 B2
`
`1
`1
`HIGH PURITY LIPOPEPTIDES
`HIGH PURITY LIPOPEPTIDES
`
`CROSS-REFERENCE TO RELATED
`CROSS-REFERENCE TO RELATED
`APPLICATIONS
`APPLICATIONS
`
`TECHNICAL FIELD OF THE INVENTION
`TECHNICAL FIELD OF THE INVENTION
`
`BACKGROUND OF THE INVENTION
`BACKGROUND OF THE INVENTION
`
`The present application is a continuation of U.S. patent
`The present application is a continuation of US. patent
`application No. 11/739,180, filed Apr. 24, 2007, now U.S.
`application No. 11/739,180, ?led Apr. 24, 2007, now US.
`Pat. No. 8,058,238, which is a continuation of U.S. patent
`Pat. No. 8,058,238, Which is a continuation of US. patent
`application Ser. No. 10/747,485, filed Dec. 29, 2003, now
`application Ser. No. 10/747,485, ?led Dec. 29, 2003, noW
`abandoned, which is a continuation of U.S. patent application 10
`abandoned, Which is a continuation of US. patent application
`Ser. No. 09/735,191 filed Nov. 28, 2000, now U.S. Pat. No.
`Ser. No. 09/735,191 ?led Nov. 28, 2000, now US. Pat. No.
`6,696,412, which claims the benefit of U.S. Provisional
`6,696,412, Which claims the bene?t of US. Provisional
`Application No. 60/177,170, filed Jan. 20, 2000, all of which
`Application No. 60/177,170, ?led Jan. 20, 2000, all of Which
`are incorporated by reference herein in their entireties.
`are incorporated by reference herein in their entireties.
`
`2
`2
`compound is currently being developed in a variety of formu­
`compound is currently being developed in a variety of formu
`lations to treat serious infections caused by bacteria, includ­
`lations to treat serious infections caused by bacteria, includ
`ing, but not limited to, methicillin resistant Staphylococcus
`ing, but not limited to, methicillin resistant Staphylococcus
`aureus (MRSA) and vancomycin resistant enterococci
`aureus (MRSA) and vancomycin resistant enterococci
`5 (VRE).
`(V RE).
`A number of United States patents describe A-21978C
`A number of United States patents describe A-21978C
`antibiotics and derivatives thereof including daptomycin (LY
`antibiotics and derivatives thereof including daptomycin (LY
`146032) as well as methods of producing and isolating the
`146032) as Well as methods of producing and isolating the
`A-21978C antibiotics and derivatives thereof.
`A-21978C antibiotics and derivatives thereof.
`U.S. Pat. Nos. Re. 32,333, Re. 32,455 and 4,800,157
`US. Pat. Nos. Re. 32,333, Re. 32,455 and 4,800,157
`describe a method of synthesizing daptomycin by cultivating
`describe a method of synthesizing daptomycin by cultivating
`Streptomyces roseosporus NRL15998 under submerged
`Streptomyces roseosporus NRL15998 under submerged
`aerobic fermentation conditions. U.S. Pat. No. 4,885,243
`aerobic fermentation conditions. US. Pat. No. 4,885,243
`describes an improved method of synthesizing daptomycin
`describes an improved method of synthesiZing daptomycin
`15 by feeding a fermentation culture a decanoic fatty acid or
`by feeding a fermentation culture a decanoic fatty acid or
`ester or salt thereof.
`ester or salt thereof.
`U.S. Pat. Nos. Re. 32,310, Re. 32,311, 4,537,717, 4,482,
`US. Pat. Nos. Re. 32,310, Re. 32,311, 4,537,717, 4,482,
`487 and 4,524,135 describe methods of deacylating the
`The present invention relates to a highly purified form of
`487 and 4,524,135 describe methods of deacylating the
`The present invention relates to a highly puri?ed form of
`A-21978C antibiotic and reacylating the peptide nucleus and
`A-21978C antibiotic and reacylating the peptide nucleus and
`lipopeptides, including daptomycin, a lipopeptide antibiotic
`lipopeptides, including daptomycin, a lipopeptide antibiotic
`with potent bactericidal activity against gram-positive bacte- 20 antibiotic derivatives made by this process. All of these pat-
`antibiotic derivatives made by this process. All of these pat
`With potent bactericidal activity against gram-positive bacte
`20
`ents describe a purified deacylated A-21978C antibiotic
`ria, including strains that are resistant to conventional antibi­
`ents describe a puri?ed deacylated A-21978C antibiotic
`ria, including strains that are resistant to conventional antibi
`nucleus or a derivative thereof which was isolated from the
`otics. The present invention also relates to a process for pre­
`nucleus or a derivative thereof Which Was isolated from the
`otics. The present invention also relates to a process for pre
`fermentation broth by filtration and then purified by Diaion
`fermentation broth by ?ltration and then puri?ed by Diaion
`paring the highly purified form of the lipopeptide. The present
`paring the highly puri?ed form of the lipopeptide. The present
`HP-20 chromatography and silica gel/C18 chromatography.
`HP-20 chromatography and silica gel/C18 chromatography.
`invention further relates to micelles of lipopeptides. The
`invention further relates to micelles of lipopeptides. The
`U.S. Pat. Nos. Re. 32,333 and Re. 32,455 disclose a puri­
`present invention also relates to pharmaceutical compositions 25
`US. Pat. Nos. Re. 32,333 and Re. 32,455 disclose a puri
`present invention also relates to pharmaceutical compositions
`25
`fication method in which a filtrate of whole fermentation
`of the lipopeptide micelles and methods of using these com­
`?cation method in Which a ?ltrate of Whole fermentation
`of the lipopeptide micelles and methods of using these com
`broth was purified through a number of precipitation and
`positions. The present invention also relates to methods of
`broth Was puri?ed through a number of precipitation and
`positions. The present invention also relates to methods of
`extraction steps to obtain a crude A-21978C complex. The
`making lipopeptide micelles from non-associated monomers
`extraction steps to obtain a crude A-21978C complex. The
`making lipopeptide micelles from non-associated monomers
`crude complex was further purified by ion exchange chroma-
`crude complex Was further puri?ed by ion exchange chroma
`of the lipopeptides, and for converting lipopeptide micelles to
`of the lipopeptides, and for converting lipopeptide micelles to
`non-associated monomers. The present invention also relates 30 tography on IRA-68 and two rounds of silica gel chromatog-
`tography on IRA-68 and tWo rounds of silica gel chromatog
`non-associated monomers. The present invention also relates
`30
`raphy. Individual A-21978C factors were separated by
`raphy. Individual A-21978C factors Were separated by
`to a process for preparing lipopeptides using micelles that is
`to a process for preparing lipopeptides using micelles that is
`reverse-phase silica gel or silica gel/C18. U.S. Pat. Nos. Re.
`reverse-phase silica gel or silica gel/C18. US. Pat. Nos. Re.
`easily scaled for commercial production.
`easily scaled for commercial production.
`32,333 and Re. 32,455 also disclose that A-21978C may be
`32,333 and Re. 32,455 also disclose that A-21978C may be
`purified by batch chromatography using Diaion HP-20 resin
`puri?ed by batch chromatography using Diaion HP-20 resin
`35 followed by silica-gel column chromatography.
`folloWed by silica-gel column chromatography.
`35
`U.S. Pat. No. 4,874,843 describes a daptomycin purifica­
`The rapid increase in the incidence of gram-positive infec­
`The rapid increase in the incidence of gram-positive infec
`US. Pat. No. 4,874,843 describes a daptomycin puri?ca
`tion method in which the fermentation broth was filtered and
`tions—including those caused by antibiotic resistant bacte­
`tionsiincluding those caused by antibiotic resistant bacte
`tion method in Which the fermentation broth Was ?ltered and
`ria—has sparked renewed interest in the development of
`passed through a column containing HP-20 resin. After elu-
`riaihas sparked reneWed interest in the development of
`passed through a column containing HP-20 resin. After elu
`novel classes of antibiotics. One such class is the lipopeptide
`tion, the semipurified daptomycin was passed through a col-
`tion, the semipuri?ed daptomycin Was passed through a col
`novel classes of antibiotics. One such class is the lipopeptide
`antibiotics, which includes daptomycin. Daptomycin has 40 umn containing HP-20ss, and then separated again on HP-20
`antibiotics, Which includes daptomycin. Daptomycin has
`umn containing HP-20ss, and then separated again on HP-20
`40
`potent bactericidal activity in vitro against clinically relevant
`resin. The '843 patent states that final resolution and separa­
`potent bactericidal activity in vitro against clinically relevant
`resin. The ’843 patent states that ?nal resolution and separa
`gram-positive bacteria that cause serious and life-threatening
`tion of daptomycin from structurally similar compounds by
`tion of daptomycin from structurally similar compounds by
`gram-positive bacteria that cause serious and life-threatening
`diseases. These bacteria include resistant pathogens, such as
`this method is impeded by the presence of impurities that are
`this method is impeded by the presence of impurities that are
`diseases. These