United States Patent [19]
`United States Patent [19]
`Baker et al. (cid:9)
`Baker et al.
`
`1111111111111111111111111111)1!191111212111111111111111111111111111111
`
`US005912226A
`[11] Patent Number: (cid:9)
`[11] Patent Number:
`[45] Date of Patent: (cid:9)
`[45] Date of Patent:
`
`5,912,226
`5,912,226
`Jun. 15, 1999
`Jun. 15, 1999
`
`[54] ANHYDRO- AND ISOMER-A-21978C CYCLIC
`[54] ANHYDRO- AND ISOMER-A-21978C CYCLIC
`PEPTIDES
`PEPTIDES
`
`[75] Inventors: Patrick J. Baker, Greenwood; Manuel
`[75] Inventors: Patrick J. Baker, Greenwood; Manuel
`Debono, Indianapolis; Khadiga Z.
`Debono, Indianapolis; Khadiga Z.
`Farid, Lebanon; R. Michael M0ll0y,
`Farid, Lebanon; R. Michael Molloy,
`Danville, all of Ind.
`Danville, all of Ind.
`
`[73] Assignee: Eli Lilly and Company, Indianapolis,
`[73] Assignee: Eli Lilly and Company, Indianapolis,
`Ind.
`Ind.
`
`[21] Appl. No.: 07/809,039
`[21] Appl. No.: 07/809,039
`[22]
`Filed:
`Dec. 16, 1991
`[22] Filed: (cid:9)
`Dec. 16, 1991
`
`Related US. Application Data
`Related U.S. Application Data
`
`[63] Continuation of application No. 07/670,375, Mar. 14, 1991,
`[63] Continuation of application No. 07/670,375, Mar. 14, 1991,
`abandoned, which is a continuation of application No.
`abandoned, which is a continuation of application No.
`07/060,148, Jun. 10, 1987, abandoned.
`07/060,148, Jun. 10, 1987, abandoned.
`[51] Int. C1.6
` A61K 38/12; C07K 11/00
`[51] Int. Cl.6 .......................... .. A61K 38/12; C07K 11/00
`[52] U.S. Cl. (cid:9)
` 514/9; 514/2; 530/317
`[52] US. Cl. .................................. .. 514/9; 514/2; 530/317
`[58] Field of Search (cid:9)
` 514/9, 2; 530/317
`[58] Field of Search ........................... .. 514/9, 2; 530/317
`
`[56]
`[56] (cid:9)
`
`References Cited
`References Cited
`
`U.S. PATENT DOCUMENTS
`U.S. PATENT DOCUMENTS
`
`Re. 32,311 12/1986 Debono (cid:9)
` 530/317
`Re. 32,311 12/1986 Debono ................................. .. 530/317
`3,953,415 4/1976 Kistaludy et al. (cid:9)
` 530/306
`3,953,415
`4/1976 Kistaludy et al.
`530/306
`4,399,067 8/1983 Debono (cid:9)
` 530/323
`4,399,067
`8/1983 Debono .......... ..
`530/323
`4,537,717 8/1985 Abbott et al. (cid:9)
` 530/317
`4,537,717
`8/1985 Abbott et al. ......................... .. 530/317
`
`OTHER PUBLICATIONS
`OTHER PUBLICATIONS
`
`Goodman et al, American Chemical Society, vol. 12, No. 1,
`Goodman et al, American Chemical Society, vol. 12, No. 1,
`(Jan. 1979), pp. 1-7.
`(Jan. 1979), pp. 1—7.
`Organic Chemistry, 3rd ed., Morrison & Boyd, p. 225.
`Organic Chemistry, 3rd ed., Morrison & Boyd, p. 225.
`
`T. Geiger et al., "Deamidation, Isomerization, and Racem-
`T. Geiger et al., “Deamidation, IsomeriZation, and Racem
`iZation at Asparaginyl and Aspartyl Residues in Peptides,”J.
`ization at Asparaginyl and Aspartyl Residues in Peptides," J.
`Biol. Chem. 262 (2), 785—794 (1987).
`Biol. Chem. 262 (2), 785-794 (1987).
`M. Bodansky et al., “Side Reactions in Peptide Synthesis,”
`M. Bodansky et al., "Side Reactions in Peptide Synthesis,"
`Synthesis 1981 (May), 333—338, 351—356.
`Synthesis 1981 (May), 333-338,351-356.
`E. A. Hagan et al., “Synthesis of Ac—Asp—Gly—Ser and
`E. A. Hagan et al., "Synthesis of Ac—Asp—Gly—Ser and
`Ac—Asp—Pro—Leu—Gly—NH2,” Int. J. Peptide Protein Res.
`Ac—Asp—Pro—Leu—Gly—NH2," Int. J. Peptide Protein Res.
`23, 642—649 (1984).
`23, 642-649 (1984).
`M. BodansZky et al., “Side Reactions in Peptide Synthesis,”
`M. Bodanszky et al., "Side Reactions in Peptide Synthesis,"
`Int. J. Peptide Protein Res. 12, 69—74 (1978).
`Int. J. Peptide Protein Res. 12, 69-74 (1978).
`B. A. Johnson et al., “Enzymatic Protein CarboXyl Methy
`B. A. Johnson et al., "Enzymatic Protein Carboxyl Methy-
`lation at Physiological pH: Cyclic Imide Formation Explains
`lation at Physiological pH: Cyclic Imide Formation Explains
`Rapid Methyl Turnover,” Biochemistry 24, 2581—2586
`Rapid Methyl Turnover," Biochemistry 24, 2581-2586
`(1985).
`(1985).
`Primary Examiner—Ponnathapura Achutamurthy
`Primary Examiner—Ponnathapura Achutamurthy
`Assistant Examiner—T. D. Wessendorf
`Assistant Examiner—T. D. Wessendorf
`Attorney, Agent, or Firm—Janet T. McClain
`Attorney, Agent, or Firm—Janet T. McClain
`
`[57]
`[57] (cid:9)
`
`ABSTRACT
`ABSTRACT
`
`TWo neW groups of A-21978C cyclic peptides, anhydro- and
`Two new groups of A-21978C cyclic peptides, anhydro- and
`isomer-A21978C peptide derivatives, have antibacterial
`isomer-A21978C peptide derivatives, have antibacterial
`activity and are useful as intermediates. The two groups are
`activity and are useful as intermediates. The tWo groups are
`prepared via transpeptidation of the parent cyclic peptides.
`prepared via transpeptidation of the parent cyclic peptides.
`Pharmaceutical formulations containing the new peptides as
`Pharmaceutical formulations containing the neW peptides as
`active ingredients and methods of treating infections caused
`active ingredients and methods of treating infections caused
`by susceptible Gram-positive bacteria With the formulations
`by susceptible Gram-positive bacteria with the formulations
`are also provided.
`are also provided.
`
`The invention also provides an antibacterial composition
`The invention also provides an antibacterial composition
`containing the new drug substance LY 146032 in substan-
`containing the neW drug substance LY 146032 in substan
`tially pure form.
`tially pure form.
`
`15 Claims, N0 Drawings
`15 Claims, No Drawings
`
`
`1 of 9
`
`FRESENIUS-KABI, Exh. 1010
`
`(cid:9)
`(cid:9)
`

`
`5,912,226
`5,912,226
`
`1
`1
`ANHYDRO- AND ISOMER-A-21978C CYCLIC
`ANHYDRO- AND ISOMER-A-21978C CYCLIC
`PEPTIDES
`PEPTIDES
`
`This application is a continuation of application Ser. No.
`This application is a continuation of application Ser. No.
`07/670,375, filed on Mar. 14, 1991, abandoned which is a 5
`07/670,375, ?led on Mar. 14, 1991, abandoned Which is a
`continuation of application Ser. No. 07/060,148, filed on
`continuation of application Ser. No. 07/060,148, ?led on
`Jun. 10, 1987, abandoned.
`Jun. 10, 1987, abandoned.
`
`SUMMARY OF THE INVENTION
`SUMMARY OF THE INVENTION
`This invention relates to two new groups of derivatives of 10
`10
`This invention relates to tWo neW groups of derivatives of
`A-21978C cyclic peptides, designated “anhydro-A-21978C
`A-21978C cyclic peptides, designated "anhydro-A-21978C
`peptide derivatives” (formula 1 compounds) and “isomer
`peptide derivatives" (formula 1 compounds) and "isomer-
`A-21978C peptide derivatives” formula 2 compounds). Like
`A-21978C peptide derivatives" formula 2 compounds). Like
`the previously knoWn A-21978C cyclic peptide derivatives
`the previously known A-21978C cyclic peptide derivatives
`(the parent cyclic peptides), the two new groups of deriva- 15
`(the parent cyclic peptides), the tWo neW groups of deriva
`15
`tives and their salts are useful semi-synthetic antibacterial
`tives and their salts are useful semi-synthetic antibacterial
`agents or are intermediates to such agents.
`agents or are intermediates to such agents.
`This invention also provides processes for preparing the
`This invention also provides processes for preparing the
`anhydro- and isomer-derivatives by trans-peptidation of the
`anhydro- and isomer-derivatives by trans-peptidation of the
`parent peptides. (cid:9)
`parent peptides.
`In another aspect, this invention provides an improved
`In another aspect, this invention provides an improved
`antibacterial composition comprising the neW drug sub
`antibacterial composition comprising the new drug sub-
`stance LY146032, or a pharmaceutically-acceptable salt
`stance LY146032, or a pharmaceutically-acceptable salt
`thereof, in substantially pure form.
`thereof, in substantially pure form.
`This invention further provides 1) methods of treating
`This invention further provides 1) methods of treating
`infections caused by susceptible Gram-positive bacteria
`infections caused by susceptible Gram-positive bacteria
`Which comprises administering a formula 1 or 2 compound
`which comprises administering a formula 1 or 2 compound
`to the animal to be treated, and 2) pharmaceutical formula
`to the animal to be treated, and 2) pharmaceutical formula-
`
`25
`25
`
`20
`20
`
`2
`
`Ser: serine
`Ser: serine
`Thr: threonine
`Thr: threonine
`Trp: tryptophan
`Trp: tryptophan
`t-BOC: tert-butoXycarbonyl
`t-BOC: tert-butoxycarbonyl
`CbZ: benZyloXycarbonyl
`Cbz: benzyloxycarbonyl
`DMF: dimethylformamide
`DMF: dimethylformamide
`THF: tetrahydrofuran
`THF: tetrahydrofuran
`HPLC: high performance liquid chromatography
`HPLC: high performance liquid chromatography
`NMR: 1H nuclear magnetic resonance
`NMR: 1H nuclear magnetic resonance
`TLC: thin-layer chromatography
`TLC: thin-layer chromatography
`UV: ultraviolet
`UV: ultraviolet
`Despite the availability of antibacterial agents today, the
`Despite the availability of antibacterial agents today, the
`need for improved antibiotics continues. Antibiotics differ in
`need for improved antibiotics continues. Antibiotics differ in
`their effectiveness against speci?c pathogenic organisms. In
`their effectiveness against specific pathogenic organisms. In
`addition, organism strains resistant to knoWn antibiotics
`addition, organism strains resistant to known antibiotics
`continue to develop. Furthermore, individual patients fre
`continue to develop. Furthermore, individual patients fre-
`quently suffer serious reactions to speci?c antibiotics, due to
`quently suffer serious reactions to specific antibiotics, due to
`hypersensitivity and/or to toxic effects. There is, therefore, a
`hypersensitivity and/or to toXic effects. There is, therefore, a
`continuing need for neW and improved antibiotics.
`continuing need for new and improved antibiotics.
`This invention relates to new antibiotics and an improved
`This invention relates to neW antibiotics and an improved
`form of the known antibiotic LY146032, which inhibit the
`form of the knoWn antibiotic LY146032, Which inhibit the
`groWth of Gram-positive bacteria. In particular, the inven
`growth of Gram-positive bacteria. In particular, the inven-
`tion relates to two new groups of A-21978C cyclic peptide
`tion relates to tWo neW groups of A-21978C cyclic peptide
`derivatives. The ?rst group of derivatives, the anhydro-A
`derivatives. The first group of derivatives, the anhydro-A-
`21978C peptide derivatives, are compounds Which have
`21978C peptide derivatives, are compounds which have
`formula 1:
`formula 1:
`
`R
`R
`
`N-R4 0
`N — R4
`
`g
`
`o
`
`H
`N
`
`\ O
`N
`H
`
`NH2
`NHZ
`
`o
`
`OH
`OH
`
`0 0
`
`HN
`HN
`
`H
`H
`N
`N
`g/Y
`N
`H
`
`O
`
`N
`
`H
`
`O
`
`O
`
`O
`O
`
`O
`
`0
`
`N
`H
`
`N
`H
`
`H
`
`0|
`
`R2
`
`O
`
`OH
`
`N
`
`H
`
`O
`
`o
`
`0
`
`N
`
`R1- N
`R1— N
`
`IL.
`
`R3
`
`N
`H
`
`o
`
`HO
`HO
`
`o
`
`tions comprising a formula 1 or 2 compound or LY146032
`tions comprising a formula 1 or 2 compound or LY146032
`in a pharmaceutically purified form as the active ingredient.
`in a pharmaceutically puri?ed form as the active ingredient.
`
`DETAILED DESCRIPTION OF THE
`DETAILED DESCRIPTION OF THE
`INVENTION
`INVENTION
`
`In this specification the following abbreviations, most of
`In this speci?cation the folloWing abbreviations, most of
`which are commonly known in the art, are used:
`Which are commonly knoWn in the art, are used:
`Ala: alanine
`Ala: alanine
`Asn: asparagine
`Asn: asparagine
`Asp: aspartic acid
`Asp: aspartic acid
`Gly: glycine
`Gly: glycine
`Kyn: kynurenine
`Kyn: kynurenine
`3-MG: L-threo-3-methylglutamic acid
`3-MG: L-threo-3-methylglutamic acid
`Orn: ornithine
`Orn: ornithine
`
`in Which R, R1 and R2 are, independently, hydrogen,
`in which R, R1 and R2 are, independently, hydrogen,
`C4—C14-alkyl, optionally substituted C2—C19-alkanoyl,
`C4—C14-alkyl, optionally substituted C2—C19-alkanoyl,
`55 C5—C19-alkenoyl or an amino-protecting group; R3, R4 and
`C5—C19-alkenoyl or an amino-protecting group; R3, R4 and
`55
`R5 are hydrogen or (i) R3 and R1 and/or (ii) R4 and R and/or
`R5 are hydrogen or
`R3 and R1 and/or (ii) R4 and R and/or
`(iii) R5 and R2, taken together, may represent a C4—C14
`(iii) R5 and R2, taken together, may represent a C4—C14
`alkylidene group; provided that 1) at least one of R, R1 or R2
`alkylidene group; provided that 1) at least one of R, R1 or R2
`must be other than hydrogen or an amino-protecting group,
`must be other than hydrogen or an amino-protecting group,
`60
`60 2) at least one of R1 or R2 must be hydrogen or an
`2) at least one of R1 or R2 must be hydrogen or an
`amino-protecting group, and 3) the R, R1 and R2 groups
`amino-protecting group, and 3) the R, R1 and R2 groups
`must together contain at least four carbon atoms; and their
`must together contain at least four carbon atoms; and their
`salts.
`salts.
`The second group of A-21978C cyclic peptide
`The second group of A-21978C cyclic peptide
`derivatives, the isomer-A-21978C peptide derivatives, are
`derivatives, the isomer-A-21978C peptide derivatives, are
`compounds Which have formula 2:
`compounds which have formula 2:
`
`65 (cid:9)
`65
`
`
`2 of 9
`
`

`
`3
`
`R
`
`N — R4
`
`O
`
`H
`N
`
`o
`
`N
`H
`
`o
`
`5,912,226
`5,912,226
`
`4
`
`OH
`OH
`
`H
`N
`
`NH
`2
`
`o o
`
`o
`
`H
`N
`
`HN
`
`O
`
`O
`
`o
`
`O
`
`H
`o N
`
`N
`H
`
`N
`H
`
`O
`
`0
`
`1
`
`R —N
`|
`R3
`R3
`
`O
`
`HO
`
`0
`
`N
`H
`
`N
`H
`
`R2
`
`N
`
`OH
`OH
`
`N
`
`0
`o
`
`HYH
`0
`O
`
`in Which R, R1, R2, R3, R4 and R5 are as de?ned supra With
`in which R, R1, R2, R3, R4 and R5 are as defined supra with
`the same provisos; and their salts.
`the same provisos; and their salts.
`The term "C4—C14-alkylidenyl" refers to a group of the
`The term “C4—C14-alkylidenyl” refers to a group of the
`formula
`formula
`
`R33
`R3 \
`
`c
`/
`R4a
`
`Greene, John Wiley and Sons, NeW York, 1981, chapter 7.
`Greene, John Wiley and Sons, New York, 1981, chapter 7.
`Especially preferably amino-protecting groups are the tert
`25 Especially preferably amino-protecting groups are the tert-
`butoXycarbonyl and benZyloXycarbonyl groups.
`butoxycarbonyl and benzyloxycarbonyl groups.
`In subgeneric aspects, the invention contemplates the
`In subgeneric aspects, the invention contemplates the
`folloWing preferred embodiments of the compound of for
`following preferred embodiments of the compound of for-
`mulas 1 and 2
`mulas 1 and 2
`(a) The compounds Wherein R is alkanoyl of the formula
`(a) The compounds wherein R is alkanoyl of the formula
`
`30
`
`40
`
`wherein R3' and R4' are hydrogen or an alkyl croup of from
`Wherein R3“ and R4“ are hydrogen or an alkyl group of from
`3 to 13 carbon atoms, provided that one of R3 and R4 must 35
`3 to 13 carbon atoms, provided that one of R3 and R4“ must
`35
`be other than hydrogen and further provided that the sum of
`be other than hydrogen and further provided that the sum of
`the carbon atoms in R3, and R4 must be no greater than 13.
`the carbon atoms in R3“ and R4“ must be no greater than 13.
`Those compounds wherein one of R and R4, R1 and R3 or
`Those compounds Wherein one of R and R4, R1 and R3 or
`R2 and R5 is C4—C14-alkylidenyl are known as Schiff's
`R2 and R5 is C4—C14-alkylidenyl are knoWn as Schiff’s
`bases. (cid:9)
`bases.
`The term "C4—C14-alkyl" refers to a univalent saturated,
`The term “C4—C14-alkyl” refers to a univalent saturated,
`straight- or branched-chain alkyl group containing from 4 to
`straight- or branched-chain alkyl group containing from 4 to
`14 carbon atoms. Those compounds wherein one of R, R1 or
`14 carbon atoms. Those compounds Wherein one of R, R1 or
`R2 are C4—C14-alkyl, referred to herein as "reduced Schiff's
`R2 are C4—C14-alkyl, referred to herein as “reduced Schiff’s
`bases”, are prepared by reduction of the corresponding
`bases", are prepared by reduction of the corresponding 45
`45
`compounds Where R and R4, R1 and R3 or R2 and R5
`compounds where R and R4, R1 and R3 or R2 and R5
`represent a C4—C14-alkylidenyl group.
`represent a C4—C14-alkylidenyl group.
`The terms “optionally substituted C2—C19-alkanoyl” and
`The terms "optionally substituted C2—C19-alkanoyl" and
`"C5-C19-alkenoyl" refer to acyl groups derived from car-
`“C5—C19-alkenoyl” refer to acyl groups derived from car
`boxylic acids containing from 2 to 19 and 5 to 19 carbon 50
`boXylic acids containing from 2 to 19 and 5 to 19 carbon
`atoms, respectively. When the group is alkanoyl, the alkyl
`atoms, respectively. When the group is alkanoyl, the alkyl
`portion is a univalent saturated, straight-chain or branched-
`portion is a univalent saturated, straight-chain or branched
`chain hydrocarbon radical which can optionally bear one
`chain hydrocarbon radical Which can optionally bear one
`hydroxyl, carboxyl, or C1—C3-alkoxy group or from one to
`hydroXyl, carboXyl, or C1—C3-alkoXy group or from one to
`three halo substituents selected from chlorine, bromine, and 55
`three halo substituents selected from chlorine, bromine, and
`55
`?uorine. When R is alkenoyl, the alkenyl portion is a
`fluorine. When R is alkenoyl, the alkenyl portion is a
`univalent, unsaturated, straight-chain or branched-chain
`univalent, unsaturated, straight-chain or branched-chain
`hydrocarbon radical containing not more than three double
`hydrocarbon radical containing not more than three double
`bonds. The double bond portion(s) of the unsaturated hydro
`bonds. The double bond portion(s) of the unsaturated hydro-
`carbon chain may be either in the cis or trans configuration. 60
`carbon chain may be either in the cis or trans con?guration.
`The term "amino-protecting group" refers to a recgonized
`The term “amino-protecting group” refers to a recgoniZed
`amino-protecting group Which is compatible With the other
`amino-protecting group which is compatible with the other
`functional groups in the A-21978C molecule. Preferably,
`functional groups in the A-21978C molecule. Preferably,
`amino-protecting groups are those which can be readily
`amino-protecting groups are those Which can be readily
`removed from the subsequently acylated compound.
`removed from the subsequently acylated compound.
`Examples of suitable protecting groups can be found in
`Examples of suitable protecting groups can be found in
`“Protective Groups in Organic Synthesis” by Theodora W.
`"Protective Groups in Organic Synthesis" by Theodora W.
`
`65 (cid:9)
`65
`
`0
`1)1
`CH3(CH2)„ — C —,
`
`wherein n is an integer from 3 to 17;
`Wherein n is an integer from 3 to 17;
`(b) The compounds Wherein R is alkanoyl of the formula
`(b) The compounds wherein R is alkanoyl of the formula
`
`0
`1)1
`CH3(CH2)„ — C —,
`CH3(CH2)11 — C — ,
`
`wherein n is 5 to 14;
`Wherein n is 5 to 14;
`(c) The compounds Wherein R is alkanoyl of the formula
`(c) The compounds wherein R is alkanoyl of the formula
`
`'1{3
`
`1)1
`CH3(CH2)„CH(CH2)„, — C — ,
`
`wherein n and m are each, independently, an integer
`Wherein n and m are each, independently, an integer
`from 0 to 14, provided that n+m must be no less than
`from 0 to 14, provided that n+m must be no less than
`1 and no greater than 15; and further provided that,
`1 and no greater than 15; and further provided that,
`when n is 0, m cannot be 8 and, when n is 1, m cannot
`When n is 0, m cannot be 8 and, When n is 1, m cannot
`be 6 or 8;
`be 6 or 8;
`(d) The compounds wherein R is cis or trans alkenyl of the
`(d) The compounds Wherein R is cis or trans alkenyl of the
`formula
`formula
`
`O
`0
`II
`CH3(CH2)„CH = CH(CH2)„,— C —
`
`wherein n and m are each, independently, an integer
`Wherein n and m are each, independently, an integer
`from 0 to 14, provided that n+m must be no less than
`from 0 to 14, provided that n+m must be no less than
`1 and no greater than 15;
`1 and no greater than 15;
`(e) The compounds where R is cis or trans alkenyl of the
`(e) The compounds Where R is cis or trans alkenyl of the
`formula
`formula
`
`
`3 of 9
`
`(cid:9)
`(cid:9)
`

`
`5
`
`10 (cid:9)
`10
`
`30
`30
`
`5
`
`0
`0
`II
`CH2= CH(CH2)„C—
`CH2: CH(CH2),,C —
`
`wherein n is an integer of from 4 to 15;
`wherein n is an integer of from 4 to 15;
`(f) The compounds Where R is alkyl of the formula
`(f) The compounds where R is alkyl of the formula
`CHACH,),— and n is an integer from 5 to 12; and
`CH3(CH2)n— and n is an integer from 5 to 12; and
`(g) The compounds Wherein R is:
`(g) The compounds wherein R is:
`
`0
`O
`II
`
`CH3(CH2)5 C—
`
`O
`II
`CH3(CH2)6 C—
`CH3(CH2)6 — C —
`O
`O
`II
`
`CH3(CH2)7 — C —
`CH3(CH2)2- C—
`O
`O
`II
`CH3(CH2)8 — C—
`CH3(CH2)8 — C —
`O
`O
`II
`CH3(CH2)9 C—
`CH3(CH2)9 — C —
`O
`O
`II
`CH3(CH2)io — C—
`O
`II
`CH2= CH— (CH2)8 — C—
`
`O
`II
`CH3(CH2)ii— C—
`
`O
`II
`CH3(CH2)12 C-
`
`0
`1)1
`CH3— (CH2)3 — CFI= CH— (CH2)7 — C —
`CH3— (CH2)3 — CH = CH- (CH2)7 — c —
`
`o
`1)1
`CH3(CH2)13 C-
`
`O
`1)1
`
`CH3CH2CH(CH3) — (CH2)5 _ C —
`CH3CH2CH(CH3) - (CH2)6 - C-
`
`CH3(CH2)ii—
`CH3(CH2)11—
`
`CH3(CH2)6 -
`CH3(CH2)e —
`0H3(CH2)9
`CH3(CH2)9 —
`
`(h) The compounds Wherein R and R4 together are:
`(h) The compounds wherein R and R4 together are:
`
`cH3(cH2)100cH—
`CH3(CH2)10OCH:
`cH3(cH2)5cH—
`CH3(CH2)5CH:
`cH3 (cH2),cH=
`
`The compounds of the formulas 1 and 2 are capable of
`The compounds of the formulas 1 and 2 are capable of
`forming salts. These salts are also part of this invention.
`forming salts. These salts are also part of this invention.
`Such salts are useful, for example, for separating and
`Such salts are useful, for example, for separating and
`purifying the compounds.
`purifying the compounds.
`For example, the compounds of formulas 1 and 2 have
`For example, the compounds of formulas 1 and 2 have
`several free carboxyl groups which can form salts. Partial,
`several free carboxyl groups Which can form salts. Partial,
`
`5,912,226
`5,912,226
`
`6
`6
`mixed and complete salts of these carboxyl groups are,
`mixed and complete salts of these carboxyl groups are,
`therefore, contemplated as part of this invention. In prepar-
`therefore, contemplated as part of this invention. In prepar
`ing these salts, pH levels greater than 10 should be avoided
`ing these salts, pH levels greater than 10 should be avoided
`due to the instability of the compounds at such levels.
`due to the instability of the compounds at such levels.
`Representative and suitable alkali-metal and alkaline
`Representative and suitable alkali-metal and alkaline-
`earth metal salts of the compounds of formulas 1 and 2
`earth metal salts of the compounds of formulas 1 and 2
`include the sodium, potassium, lithium, cesium, rubidium,
`include the sodium, potassium, lithium, cesium, rubidium,
`barium, calcium and magnesium salts.
`barium, calcium and magnesium salts.
`The alkali-metal and alkaline-earth-metal cationic salts of
`The alkali-metal and alkaline-earth-metal cationic salts of
`the compounds for formula 1 and 2 are prepared according
`the compounds for formula 1 and 2 are prepared according
`to procedures commonly used for the preparation of cationic
`to procedures commonly used for the preparation of cationic
`salts. For example, the free acid form of a formula 1 or 2
`salts. For example, the free acid form of a formula 1 or 2
`15 compound is dissolved in a suitable solvent such as warm
`compound is dissolved in a suitable solvent such as Warm
`15
`methanol or ethanol. A solution containing a stoichiometric
`methanol or ethanol. A solution containing a stoichiometric
`quantity of the desired inorganic base in aqueous methanol
`quantity of the desired inorganic base in aqueous methanol
`is added to this solution. The salt thus formed can be isolated
`is added to this solution. The salt thus formed can be isolated
`by routine methods, such as filtration or evaporation of the
`by routine methods, such as ?ltration or evaporation of the
`solvent. Aconvenient method of preparing salts is by the use
`20 solvent. A convenient method of preparing salts is by the use
`20
`of ion-exchange resins.
`of ion-exchange resins.
`Suitable amine salts of the formula 1 and 2 compounds
`Suitable amine salts of the formula 1 and 2 compounds
`include the ammonium and the primary, secondary, and
`include the ammonium and the primary, secondary, and
`tertiary C1—C4-alkylammonium and hydroxy-C2—C4
`tertiary C1—C4-alkylammonium and hydroxy-C2—C4-
`25
`alkylammonium salts. Illustrative amine salts include those
`25 alkylammonium salts. Illustrative amine salts include those
`formed by reaction of a formula 1 or 2 compound with
`formed by reaction of a formula 1 or 2 compound With
`ammonium hydroxide, methylamine, sec-butylamine,
`ammonium hydroxide, methylamine, sec-butylamine,
`isopropylamine, diethylamine, di-isopropylamine,
`isopropylamine, diethylamine, di-isopropylamine,
`cyclohexylamine, ethanolamine, triethylamine, 3-amino-1
`cyclohexylamine, ethanolamine, triethylamine, 3-amino-l-
`propanol and the like.
`propanol and the like.
`The salts formed with organic amines can also be pre-
`The salts formed With organic amines can also be pre
`pared by Well knoWn procedures. For example, the gaseous
`pared by well known procedures. For example, the gaseous
`or liquid amine can be added to a solution of a formula 1 or
`or liquid amine can be added to a solution of a formula 1 or
`35 2 compound in a suitable solvent such as ethanol. The
`2 compound in a suitable solvent such as ethanol. The
`35
`solvent and excess amine can be removed by evaporation.
`solvent and excess amine can be removed by evaporation.
`Because the compounds of this invention also have free
`Because the compounds of this invention also have free
`amino groups, they can, therefore, form acid addition salts.
`amino groups, they can, therefore, form acid addition salts.
`Such salts are also part of this invention. Representative and
`Such salts are also part of this invention. Representative and
`40
`40 suitable acid-addition salts of the compounds of formula 1 or
`suitable acid-addition salts of the compounds of formula 1 or
`2 include those salts formed by standard reaction with both
`2 include those salts formed by standard reaction With both
`organic and inorganic acids such as, for example,
`organic and inorganic acids such as, for example,
`hydrochloric, sulfuric, phosphoric, acetic, succinic, citric,
`hydrochloric, sulfuric, phosphoric, acetic, succinic, citric,
`lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic,
`lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic,
`D-glutamic, d-camphoric, glutaric, glycolic, phthalic,
`D-glutamic, d-camphoric, glutaric, glycolic, phthalic,
`tartaric, lauric, stearic, salicylic, methanesulfonic,
`tartaric, lauric, stearic, salicylic, methanesulfonic,
`benzenesulfonic, sorbic, picric, benzoic, cinnamic and like
`benZenesulfonic, sorbic, picric, benZoic, cinnamic and like
`acids.
`acids.
`Pharmaceutically acceptable alkali-metal, alkaline-earth
`Pharmaceutically acceptable alkali-metal, alkaline-earth-
`metal, amine and acid-addition salts are a particularly useful
`metal, amine and acid-addition salts are a particularly useful
`group of compounds of this invention.
`group of compounds of this invention.
`The formula 1 and 2 compounds are prepared from
`The formula 1 and 2 compounds are prepared from
`previously knoWn A-21978C cyclic peptides, Which in turn
`previously known A-21978C cyclic peptides, which in turn
`are prepared from the A-21978C antibiotics. The A-21978C
`are prepared from the A-21978C antibiotics. The A-21978C
`antibiotics, a group of closely related, acidic peptide
`antibiotics, a group of closely related, acidic peptide
`antibiotics, are described by Robert L. Hamill and Marvin
`antibiotics, are described by Robert L. Hamill and Marvin
`M. Hoehn in U.S. Pat. No. 4,208,403, issued Jun. 17, 1980.
`M. Hoehn in US. Pat. No. 4,208,403, issued Jun. 17, 1980.
`60 As described in U.S. Pat. No. 4,208,403, the A-21978
`As described in US. Pat. No. 4,208,403, the A-21978
`60
`antibiotic complex contains a major component, factor C,
`antibiotic complex contains a major component, factor C,
`which is itself a complex of closely related factors. A-21978
`Which is itself a complex of closely related factors. A-21978
`factor C, Which is called the A-21978C complex, contains
`factor C, which is called the A-21978C complex, contains
`individual factors C0, C1, C2, C3, C4 and C5. Factors C1, C2
`individual factors Co, C1, C2, C3, C4 and C5. Factors C1, C2
`65 and C3 are major factors; and factors Co, C4 and C, are
`and C3 are major factors; and factors C0, C4 and C5 are
`65
`minor factors. The A-21978C factors have the structure
`minor factors. The A-21978C factors have the structure
`shown in formula 3:
`shoWn in formula 3:
`
`45
`45
`
`50 (cid:9)
`50
`
`55
`55
`
`
`4 of 9
`
`

`
`8
`8
`we found that the LY146032 material contained two impu-
`We found that the LY146032 material contained tWo impu
`rities. The impurities were more pronounced when
`rities. The impurities Were more pronounced When
`LY146032 was in solution in the pH range of 4 to 6. Our
`LY146032 Was in solution in the pH range of 4 to 6. Our
`work led to the isolation of these materials and to the further
`Work led to the isolation of these materials and to the further
`discovery that they Were closely related to LY146032. Like
`5 discovery that they were closely related to LY146032. Like
`LY146032, the new compounds also have antibacterial
`LY146032, the neW compounds also have antibacterial
`activity. Identi?cation of the tWo materials and subsequent
`activity. Identification of the two materials and subsequent
`studies shoWed that they Were formed by a transpeptidation
`studies showed that they were formed by a transpeptidation
`reaction. This reaction involves 3 compounds: 1) the starting
`reaction. This reaction involves 3 compounds: 1) the starting
`ot-aspartyl peptide (LY146032), 2) a stable intermediate and
`a-aspartyl peptide (LY146032), 2) a stable intermediate and
`3) the [3-aspartyl isomer of LY146032.
`3) the 13-aspartyl isomer of LY146032.
`The stable intermediate was found to be the compound of
`The stable intermediate Was found to be the compound of
`formula 1 Wherein R is n-decanoyl and R1, R2, R3, R4 and
`formula 1 wherein R is n-decanoyl and R1, R2, R3, R4 and
`R5 are hydrogen. In discussions herein, this compound is
`R5 are hydrogen. In discussions herein, this compound is
`designated “anhydro-LY146032”.
`designated "anhydro-LY146032".
`The third compound was found to be the 13-aspartyl
`The third compound Was found to be the [3-aspartyl
`isomer of LY146032, i.e. the formula 2 compound Wherein
`isomer of LY146032, i.e. the formula 2 compound wherein
`R is n-decanoyl and R1, R2, R3, R4, and R5 are hydrogen. In
`R is n-decanoyl and R1, R2, R3, R4, and R5 are hydrogen. In
`discussions herein this compound is designated “isomer
`discussions herein this compound is designated "isomer-
`LY146032".
`LY146032”.
`Thus, the formula 1 and 2 compounds are formed by
`Thus, the formula 1 and 2 compounds are formed by
`aspartyl transpeptidation of the parent cyclic peptides, Which
`aspartyl transpeptidation of the parent cyclic peptides, which
`include LY146032. The transpeptidation involves tWo
`include LY146032. The transpeptidation involves two
`distinct, reversible steps: (1) formation of the compounds of
`distinct, reversible steps: (1) formation of the compounds of
`formula 1 (the anhydro intermediates) from either the parent
`formula 1 (the anhydro intermediates) from either the parent
`ot-aspartyl peptide or from the formula 2 peptides (the
`a-aspartyl peptide or from the formula 2 peptides (the
`[3-aspartyl peptides) and (2) hydrolysis of the intermediate
`(3-aspartyl peptides) and (2) hydrolysis of the intermediate
`formula 1 compounds to either the parent ot-aspartyl pep
`formula 1 compounds to either the parent a-aspartyl pep-
`tides or to the 13-aspartyl peptides of formula 2.
`tides or to the [3-aspartyl peptides of formula 2.
`The mechanism of transpeptidation involves formation of
`The mechanism of transpeptidation involves formation of
`a succinimide intermediate, probably through intramolecu
`a succinimide intermediate, probably through intramolecu-
`lar dehydration of the free carboXyl group of aspartic acid
`lar dehydration of the free carboxyl group of aspartic acid
`and the amino group of the neighboring glycine. This step is
`and the amino group of the neighboring glycine. This step is
`followed by nucleophilic hydroxide attack of either the a- or
`folloWed by nucleophilic hydroxide attack of either the ot- or
`[3-carbonyl of the succinimido intermediate Which results in
`35 3-carbonyl of the succinimido intermediate which results in
`35
`formation of the corresponding [3- or ot-aspartyl peptide.
`formation of the corresponding 13- or a-aspartyl peptide.
`Formation of the [3-aspartyl peptide predominates by a factor
`Formation of the 13-aspartyl peptide predominates by a factor
`of 2—3, presumably because of the greater electrophilicity of
`of 2-3, presumably because of the greater electrophilicity of
`the ot-carbonyl of the succinimide intermediate. The
`the a-carbonyl of