(19) United States
`(12) Patent Application Publication (10) Pub. No.: US 2007/0117743 A1
`Palomera et al.
`(43) Pub. Date:
`May 24, 2007
`
`US 200701 17743A1
`
`(54) NEW ANTITUMORAL COMPOUNDS
`(76) Inventors: Fernando Albericio Palomera,
`Barcelona (ES); Ariadna Fernandez
`Donis, Barcelona (ES); Ernest Giralt
`Lledo, Barcelona (ES); Carolina
`Gracia Cantador, Barcelona (ES);
`Pilar Lopez Rodriguez, Barcelona
`(ES); Sonia Varon Colomer, Barcelona
`(ES); Carmen Cuevas Marchante,
`Madrid (ES); Angel Lopez Macia,
`Madrid (ES); Andres Francesch
`Soloso, Madrid (ES); Jose-Carlos
`Jimenez Garcia, Barcelona (ES);
`Miriam Royo Exposito, Barcelona
`(ES)
`Correspondence Address:
`KING & SPALDING
`1185 AVENUE OF THE AMERICAS
`NEW YORK, NY 10036-4003 (US)
`(21) Appl. No.:
`10/570,734
`
`(22) PCT Filed:
`(86). PCT No.:
`
`Sep. 9, 2004
`PCT/GBO4/O3847
`
`S 371(c)(1),
`Oct. 18, 2006
`(2), (4) Date:
`Foreign Application Priority Data
`
`(30)
`
`Sep. 9, 2003 (GB)......................................... O321066.3
`
`Publication Classification
`
`(51) Int. Cl.
`(2006.01)
`A6II 38/12
`(2006.01)
`C07K 7/60
`(52) U.S. Cl. ................................................. 514/9; 530/317
`
`(57)
`
`ABSTRACT
`
`New analogues of kahalalide F are provided.
`
`MPI EXHIBIT 1066 PAGE 1
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`NEW ANTITUMORAL COMPOUNDS
`
`FIELD OF THE INVENTION
`0001. The present invention is directed to new kahalalide
`antitumoral compounds, in particular to analogues of
`kahalalide F, pharmaceutical compositions containing them
`and their use as antitumoral, antiviral, antifungal agents and
`in the treatment of psoriasis.
`
`BACKGROUND OF THE INVENTION
`0002 The kahalalide compounds are peptides isolated
`from a Hawaiian herbivorous marine species of mollusc,
`Elysia rufescens and its diet, the green alga Bryopsis sp.
`Kahalalide F is described in Hamann, Met al., J. Am. Chem.
`Soc., 1993, 115, 5825-5826.
`0003 Kahalalide A-G are described in Hamann, M. et al.,
`J. Org. Chem, 1996, 61, 6594-6600: “Kahalalides: bioactive
`peptides from a marine mollusk Elysia rufescens and its
`algal diet Bryopsis sp.'.
`0004) Kahalalide Hand Jare described in Scheuer P.J. et
`al., J. Nat. Prod. 1997, 60,562-567: “Two acyclic kahala
`lides from the Sacoglossan mollusk Elysia rufescens'.
`
`0005) Kahalalide O is described in Scheuer P. J. et al., J.
`Nat. Prod. 2000, 63(1) 152-4: “A new depsipeptide from the
`sacoglossan mollusk Elysia Ornata and the green alga Bry
`opsis species'.
`
`0006 For kahalalide K, see Kan, Y. et al., J. Nat. Prod.
`1999 62(8) 1169-72: “Kahalalide K: A new cyclic depsipep
`tide from the hawaiian green alga Bryopsis species'.
`0007 For related reports, see also Goetz et al., Tetrahe
`dron, 1999, 55; 7739-7746: “The absolute stereochemistry
`of Kahalalide F: Albericio, F. et al. Tetrahedron Letters,
`2000, 41,9765-9769: “Kahalalide B. Synthesis of a natural
`cyclodepsipeptide'. Becerro et al. J. Chem. Ecol. 2001,
`27(1-1), 2287-99: “Chemical-defenses of the sarcoglossan
`mollusk Elysia rufescens and its host Alga bryopsis sp.'.
`0008 Of the kahalalide compounds, kahalalide F is the
`most promising because of its antitumoral activity. Its struc
`ture is complex, comprising six amino acids as a cyclic part,
`and an exocyclic chain of seven amino acids with a terminal
`fatty acid group. Originally kahalalide F was reported to
`have the structure (I).
`
`D-allo-Ile 7
`
`D-allo-Ile 5
`
`(I)
`
`D-Val 4
`
`O
`
`cy
`
`Ple 3
`
`N- D-allo-Thr 6 N
`-
`
`H
`N
`
`H
`N
`
`N
`
`O
`
`N
`
`S.
`H
`N S
`s
`
`D-Wall 10
`
`D-Pro 9
`
`s
`s
`
`\r
`
`O
`
`Wall 11
`
`O
`Thr 12
`r
`
`O
`
`O
`
`NH
`
`HN
`
`OH
`
`O
`
`'''
`
`D-Wall 13
`
`NH
`
`O
`
`5-Mehex 14
`
`HN
`Orn 8
`
`O
`
`O
`
`HN
`
`O
`
`O
`
`Wall 1
`
`HN
`
`NH
`
`A \
`
`(Z)-Dhb 2
`
`MPI EXHIBIT 1066 PAGE 2
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`0009. In WO 04035613 there is described the 4(S)-
`methylhexyl compound with the following formula (II). WO
`04035613 is incorporated here in full by specific reference.
`
`D-allo-Ile 7
`
`D-allo-Ile 5
`
`(II)
`
`O.
`
`H.
`N
`
`O
`
`N
`
`s
`s
`
`H
`N
`
`E
`s
`
`N
`
`HN
`Orn 8
`
`NH
`
`O
`
`Thr 12
`
`OH
`
`HN
`
`O
`
`D-Wall 13
`
`v
`
`y
`
`O
`
`NH
`
`4(S)-Mehex 14
`
`D-Val 4
`
`s
`
`H
`S
`NN s
`
`1. H
`
`O
`
`HN
`
`O
`
`O
`
`NH
`
`- HN
`
`Wall 1
`
`\
`
`Ple 3
`
`O
`
`O
`
`0010. The activity of kahalalide F against in vitro cell
`cultures of human lung carcinoma A-549 and human colon
`carcinoma HT-29 were reported in EP 610 078. Kahalalide
`F has also demonstrated to have antiviral and antifungal
`properties, as well as to be useful in the treatment of
`psoriasis.
`0011 WO 0236145 describes pharmaceutical composi
`tions containing kahalalide F and new uses of this compound
`in cancer therapy and is incorporated herein by reference in
`its entirety.
`0012 WO 03 33012 describes the clinical use in oncol
`ogy of kahalalide compounds and is incorporated herein by
`reference in its entirety.
`
`0013 The synthesis and cytotoxic activities of natural
`and synthetic kahalalide compounds is described in WO 01
`58934, which is incorporated herein by reference in its
`entirety. WO 0158934 describes the synthesis of kahalalide
`F and also of compounds with a similar structure in which
`the terminal fatty acid chain is replaced by other fatty acids.
`0014. There is still a need to provide further antitumoral
`compounds, in particular further kahalalide compounds with
`improved properties.
`
`SUMMARY OF THE INVENTION
`0015 We have found kahalalide analogue compounds
`with improved biological activity.
`
`MPI EXHIBIT 1066 PAGE 3
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`0016. The present invention is directed to compounds of
`formula 1
`
`(I)
`
`D-allo-Ile 7
`
`D-allo-Ile 5
`
`N- D-allo-Thr 6
`
`O
`
`H
`
`N-
`
`O
`
`O
`
`\ ev 4
`S Y HN
`
`O
`
`NH
`
`O
`
`Wall 1
`
`HN
`
`O
`
`\
`
`Ple 3
`
`5-Mehex 14
`
`wherein one or more amino acids in the cyclic or exocyclic
`part have been substituted by other natural or non natural
`amino acids, have been masked with organic groups or have
`been removed. The present invention is also directed to
`compounds of formula 1 wherein the aliphatic terminal acid
`group has been Substituted by other acyl groups or has been
`removed.
`0017. The present invention is also directed to a pharma
`ceutical composition comprising a compound as previously
`defined and a pharmaceutically acceptable carrier, vehicle or
`diluent.
`0018. The present invention further provides a method of
`treating any mammal, notably a human, affected by cancer,
`viral infection, fungal infection or psoriasis which comprises
`administering to the affected individual a therapeutically
`effective amount of a compound as defined above.
`0019. The present invention can be employed particularly
`for treatment of patients with refractory cancers that do not
`respond favourably to other treatments. In particular, the
`
`compositions of this invention can be employed after other
`chemotherapy has been tried and not worked.
`0020. The present invention is particularly directed to the
`treatment of patients affected with prostate cancer, breast
`cancer, hepatocellular carcinoma, melanoma, colorectal can
`cer, renal cancer, ovarian cancer, NSCL cancer, epithelial
`cancer, pancreatic cancer and tumors that overexpress the
`Her2/neu oncogene.
`0021. In another aspect the present invention is directed
`to the use of a compound as defined above in the manufac
`ture of a medicament. In a preferred embodiment the medi
`cament is for the treatment of cancer, psoriasis, viral infec
`tion or fungal infection.
`0022. The invention additionally provides kits compris
`ing separate containers containing a pharmaceutical compo
`sition comprising a compound as defined above and a
`reconstituting agent. Methods of reconstitution are also
`provided.
`
`MPI EXHIBIT 1066 PAGE 4
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`DETAILED DESCRIPTION OF THE
`INVENTION
`0023. We have identified analogues of kahalalide F that
`show significant improvement in activity with respect to
`kahalalide F.
`0024. The present invention is directed to compounds of
`formula 1
`
`0028. Thus, in one aspect of the present invention, there
`are provided compounds of formula 1 wherein there is not
`L-Orn at position 8.
`0029. The L-Orn can be replaced by D-Orn, or by another
`natural or non-natural amino acid. For example, the L-Orn
`can be replaced by a natural amino acid, Such as lysine; or
`a masked natural amino acid, Such as arginine or lysine with
`
`D-Wall 10
`
`O
`
`O.
`
`H.
`N
`
`D-Pro 9
`
`s
`s
`
`H
`N
`
`N
`
`O
`
`D-allo-Ile 7
`
`N
`
`D-allo-Ile 5
`
`
`
`1.
`
`(I)
`
`N H -, Q
`
`Thr 12
`
`OH
`
`HN
`
`HN
`Orn 8
`
`O
`
`s'
`
`D-Wall 13
`
`NH
`
`5-Mehex 14
`
`wherein one or more exocyclic or cyclic amino acids have
`been Substituted by other natural or non natural amino acids,
`have been masked with organic groups or have been
`removed. The present invention is also directed to com
`pounds of formula 1 wherein the aliphatic methylhexanoic
`acyl group has been Substituted by other acyl groups or has
`been removed.
`Exocyclic Amino Acids
`0.025
`Preferred compounds of the invention include
`those of formula 1 wherein one or more amino acids of the
`exocyclic chain have been substituted by other natural or
`non natural amino acids, have been masked with organic
`groups or have been removed.
`0026.
`In particular, preferred compounds include those
`with 1, 2 or 3 replacement amino acids in the exocyclic
`chain; and those with 1, 2, 3, 4, 5 or 6 removed amino acids
`in the exocyclic chain.
`0027. Especially preferred are those compounds of for
`mula 1 wherein one exocyclic amino acid has been Substi
`tuted by another natural or non natural amino acid, and/or
`has been masked with one or more Substituent organic
`groups. This preference is specially applicable to the L-Orn
`amino acid in position 8.
`
`one or more alkyl, phenyl or oligomethylene Substituents, for
`example N(Me)N'(Me)-Arg, N(Me.Ph).N'(Me)-Arg,
`N(CH2)4N'(Me)-Arg, N(CH2)N(CH2)4-Arg, N(Me)-
`Lys.
`0030 The L-Orn can be masked. For example, the amino
`group of the L-Orn may have Substituents, notably alkyl
`substituents that may be further substituted, notably with
`heterocyclic groups,
`for example N(CHN(CH),
`N(CH2))-Orn; or more complex substituents as in biotiny
`lornithine or Orn(NTfa).
`Cyclic Amino Acids
`0031. Other preferred compounds of the invention
`include those of formula 1 wherein one or more amino acids
`of the cyclic chain have been substituted by other natural or
`non natural amino acids, have been masked with organic
`groups or have been removed.
`0032. In particular, preferred compounds include those
`with 1, 2 or 3 replacement amino acids in the cyclic chain.
`0033 Especially preferred are those compounds of for
`mula 1 wherein one or more amino acids have been Substi
`tuted by other natural or non natural amino acids, and/or
`have been masked with organic groups. This preference is
`specially applicable to the L-Phe amino acid in position 3.
`
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`0034 Thus, in one aspect of the present invention, there
`are provided compounds of formula 1 wherein the L-Phe at
`position 3 has been replaced or masked.
`0035) The L-Phe can be replaced by D-Phe, or by another
`natural or non-natural amino acid. For example, the L-Phe
`can be replaced by a natural amino acid, such as tyrosine; an
`alkyl-substituted amino acid, Such as N-methyltyrosine; an
`aryl-substituted amino acid, Such as 2-amino-3-biphenyl-4-
`yl-propionic acid, 2-amino-3-naphthalen-2-yl-propionic
`acid or aminophenylacetic acid; an heterocyclyl-substituted
`amino acid. Such as 2-amino-3-thiophen-2-ylpropionic acid;
`or a cyclic amino acid where the amino group is part of a
`heterocycle. Such as 1,2,3,4-tetraisoquinoline-3-carboxilic
`acid or octahydroisoindole-1-carboxylic acid.
`0036) The L-Phe can be masked. For example, the phenyl
`ring may be partially or fully Saturated, and may be substi
`tuted with one or more substituents. Substituents for the
`phenyl ring may be as indicated, preferably halo or nitro.
`The amino group may have 1 or 2 substituents, notably alkyl
`Such as methyl, especially 1 methyl Substituent.
`Terminal Acyl Group
`0037 Besides the modification of the amino acids of the
`exocyclic chain and/or of the cyclic part, there can also be
`compounds with a modification in the terminal acyl group of
`the exocyclic chain.
`0038 For example, the acyl group can comprise one or
`more substituents, especially aromatic, heterocyclic or car
`bocyclic groups which in turn may have one or more
`Substitutents, for example can be a benzoyl group which
`may have one or more Substituents, a phenylalkanoyl which
`may have one or more Substituents or a cinnamoyl which
`may have one or more Substituents. Additionally, the termi
`nal acyl group can be another fatty acid typically a saturated
`or unsaturated fatty acid with 3 to 26 carbon atoms, more
`especially 12 to 20 carbon atoms.
`0.039
`Typical groups for adoption in place of the terminal
`acyl group in formula 1 include:
`0040 Straight chain alkanoyl with up to 26 carbon
`atoms and with one or more substituents, especially
`substituents chosen from optionally substituted
`cycloalkyl Such as 4-methylcyclohexyl, alkyl, espe
`cially short chain alkyl Such as methyl, optionally
`substituted aryl especially phenyl such as difluorophe
`nyl; halo Such as fluoro up to perfluoro, oxo; amino; or
`imino.
`0041. Optionally substituted benzoyl such as benzoyl
`itself, p-methylbenzoyl acid, p-trifluoromethylbenzoyl
`acid, piperonyloyl.
`0042 Arylalkenoyl group with preferably two carbon
`atoms in the alkenoyl especially a cinnamoyl group
`Such as p-trifluoromethylcinnamoyl.
`0.043 Moreover, the acyl group can be replaced by
`another acyl group, preferably of formula R'CO . R' is as
`defined and is suitably alkyl, alkenyl, aryl, heterocyclyl, or
`carbocyclyl, and may itself be substituted.
`0044 Finally, there can be compounds with a unique
`modification located in the fatty acid of the exocyclic chain,
`the provision of a 5-methylhexyl terminal group.
`
`0045 Combined Changes
`0046) The possible changes in the exocyclic amino acids,
`cyclic amino acids and terminal acyl group can be taken in
`combination.
`0047 Thus, other preferred compounds of the invention
`are those of formula 1 wherein one or more amino acids of
`the exocyclic chain and/or one or more amino acids of the
`cyclic part have been modified, as mentioned above and/or
`the terminal acyl group has been modified.
`0048. The compounds where the L-Orn at position 8 has
`been replaced or masked may adopt other preferred modi
`fications, including a 4(S)-methylhexyl or other group in the
`terminal group of the sidechain.
`0049. The compounds where the L-Phe at position 3 has
`been replaced or masked may adopt other preferred modi
`fications, including a 4(S)-methylhexyl or other group in the
`terminal group of the sidechain.
`0050 Examples of substituent groups include C-Cls
`alkyl, C-Cs alkenyl, C-Cs alkynyl, aryl, heterocyclic
`groups, OR', SR', SOR', SOR, NO, NHR', N(R'),
`NHCOR', N(COR), NHSOR', CN, halogen, C(=O)R',
`COR', OC(=O)R’ wherein each of the R' groups is inde
`pendently selected from the group consisting of H, OH,
`NO, NH, SH, CN, halogen, C(=O)H, C(=O)alkyl,
`COH, substituted or unsubstituted C-Cs alkyl, substituted
`or unsubstituted C-C alkenyl, Substituted or unsubstituted
`C-Cls alkynyl and substituted or unsubstituted aryl. Suit
`able halogen Substituents in the compounds of the present
`invention include F, Cl, Brand I.
`0051 Alkyl groups have a saturated linear or branched
`hydrocarbon group including, for example, methyl, ethyl,
`isopropyl, isobutyl, t-butyl, heptyl, dodecyl, octadecyl.
`amyl, 2-ethylhexyl, 2-methylbutyl, 5-methylhexyl, 9-me
`thyl-3-decyl, and the like, particularly alkyl groups which
`have a single branched methyl group. Suitably the alkyl
`group is long and has 1 to 20 carbon atoms, more typically
`1 to 15 or 1 to 10 carbon atoms, or can be short and has 1
`to 6 or 1 to 3 carbon atoms. Long carbon chains are
`candidates for use in the terminal fatty acid group.
`0052 Preferred alkenyl and alkynyl groups in the com
`pounds of the present invention have one or more unsatur
`ated linkages and from 2 to 12 carbon atoms, more prefer
`ably 2 to 8 carbon atoms, still more preferably 2 to 6 carbon
`atoms, even more preferably 2, 3 or 4 carbon atoms. The
`terms alkenyl and alkynyl as used herein refer to both cyclic
`and noncyclic groups, although straight or branched non
`cyclic groups are generally more preferred. In a general
`sense, we include alkylidene within alkenyl, they both being
`substituents with a double bond.
`0053 Suitable aryl groups in the compounds of the
`present invention include single and multiple ring com
`pounds, including multiple ring compounds that contain
`separate and/or fused aryl groups. Typical aryl groups con
`tain from 1 to 3 separated or fused rings and from 6 to about
`18 carbon ring atoms. Specifically preferred aryl groups
`include substituted or unsubstituted phenyl, naphthyl, biphe
`nyl, phenanthryl and anthracy1.
`0054 Suitable acyl groups include alkanoyl groups
`which have from 2 to about 12 carbon atoms, more prefer
`ably from 2 to about 8 carbon atoms, still more preferably
`
`MPI EXHIBIT 1066 PAGE 6
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`from 2 to about 6 carbon atoms, even more preferably 2
`carbon atoms. Other acyl groups include alkenylacyl, alky
`nylacyl, arylacyl, heterocyclylacyl.
`0.055
`Suitable heterocyclic groups include heteroaro
`matic and heteroalicyclic groups. Suitable heteroaromatic
`groups in the compounds of the present invention contain
`one, two or three heteroatoms selected from N, O or Satoms
`and include, e.g., coumarinyl including 8-coumarinyl,
`quinolinyl including 8-quinolinyl, pyridyl, pyrazinyl, pyrim
`idyl, furyl, pyrrolyl, thienyl, thiazolyl, oxazolyl, imidazolyl,
`indolyl, benzofuranyl and benzothiazol. Suitable heteroali
`cyclic groups in the compounds of the present invention
`contain one, two or three heteroatoms selected from N, O or
`Satoms and include, e.g., tetrahydrofuranyl, tetrahydropy
`ranyl, piperidinyl, morpholino and pyrrolindinyl groups.
`0056. The groups above mentioned may be substituted at
`one or more available positions by one or more substituents.
`
`0057 The natural amino acids include alanine, glycine,
`valine, leucine, isoleucine, phenylalanine, tyrosine, tryp
`tophan, methionine, cysteine, aspartate, asparagine,
`glutamatic acid, glutamine, lysine, arginine, proline, serine,
`threonine, histidine and hydroxyproline
`0.058 We also refer to the WO 0158934 incorporated
`herein in full by specific reference. That earlier text gives
`guidance which is applicable to the compounds of this
`invention, and we import the definitions for adoption in the
`compounds of this invention.
`0059 Abbreviations used in the following description for
`amino acids and the designations of peptides follow the rules
`of the IUPAC-IUB Commission of Biochemical Nomencla
`ture in J. Biol. Chem., 1972, 247,977-983.
`0060. The following additional abbreviations are used:
`
`Alloc
`
`AM
`AO
`Bip
`Boc
`BTFFH
`
`-Bu
`BZa-OH
`Cha
`
`p-CFBza-OH
`p-CFBZAc-OH
`p-CFCinn-OH
`CI-TrtC-resin
`Dha
`Z-Dhb
`DIPEA
`DMF
`EDC.HCI
`
`Fmoc
`6,6-dFHep-OH
`3,5-dFPhAc-OH
`4-Guut-OH
`GISO
`HAPyU
`
`HATU
`
`hCh
`
`Hep-OH
`HOAC
`HOAt
`
`HOBt
`cos-OH
`LCSO
`Lit-OH
`MeHex-OH
`M2A
`
`(cft-4Me-cHexa)-OH
`
`allyloxycarbonyl
`2-amino-5-(dipyrrollidin-1-yl-methylene)-
`amino-pentanoic acid
`maslinic acid
`oleanolic acid
`2-amino-3-biphenyl-4-yl-propionic acid
`tert-butyloxycarbonyl
`bis(tetramethylene) fluoroformamidinium
`hexafluorophosphate
`tert-Butyl
`benzoic acid
`cyclohexylalanine or 2-amino-3-cyclohexyl
`propionic acid
`4-trifluoromethylbenzoic acid
`3-(4-Trifluoromethylbenzyl)acetic acid
`3-(4-trifluoromethylbenzyl)acrylic acid
`2-chlorotrityl chloride-resin
`2-aminoacrylic acid or didehydroalanine
`C,3-didehydro-C-aminobutyric acid
`N,N-diisopropylethylamine
`N,N-dimethylformamide
`1-ethyl-3-(3'-
`dimethylaminopropyl)carbodiimide
`hydrochloride
`9-fluorenylmethoxycarbonyl
`6,6-difluoro-heptanoic acid
`(3,5-difluorophenyl)acetic acid
`4-guanidinobutyric acid
`growth inhibition at 50%
`O-(7-azabenzotriazol-1-yl)-1,1,3,3-
`bis(tetramethylene)uronium
`hexafluorophosphate
`O-(7-azabenzotriazol-1-yl)-1,1,3,3-
`etramethyluronium hexafluorophosphate
`homocyclohexylalanine or 2-amino-4-
`cyclohexylbutyric acid
`heptanoic acid
`acetic acid
`-hydroxy-7-azabenzotriazole (3-hydroxy
`3H-1,2,3-triazolo-4,5-bipyridine)
`-hydroxybenzotriazole
`icosanoic acid
`ethal concentration at 50%
`itocholic acid
`methylhexanoic acid
`(tetramethylene)-1H-1,2,3-triazolo4,5-
`bipyridino-1-ylmethylene-N-
`methylmethanaminium hexafluorophosphate
`(cis/trans)-4-methylcyclohexanecarboxylic
`acid
`
`MPI EXHIBIT 1066 PAGE 7
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`US 2007/01 17743 A1
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`
`-continued
`
`4-Methyl-benzoic acid
`2-Amino-5-(N',N',N',N'-
`tetramethylguanidino)pentanoic acid
`2-amino-5-(dimethylamino-pyrrollidin-1-yl
`methylene)-amino-pentanoic acid
`methanol
`
`guanidino)pentanoic acid
`N-methylmorpholine
`myristic acid or tetradecanoic acid
`2-amino-3-naphthalen-2-yl-propionic acid
`octanoic acid
`6-oxo-heptanoic acid
`octahydro-isoindole-1-carboxylic acid
`2-amino-5-(dipyrrollidin-1-yl-methyl)-
`aminopentanoic acid
`perfluoroheptanoic acid
`phenylglycine or aminophenylacetic acid
`5-(R)-phenyl-pirrollidine-2-carboxilic acid
`pipecolic acid
`benzo1,3dioxole-5-carboxilic acid
`Solid-phase synthesis
`trifluoroacetyl
`trifluoroacetic acid
`trifluoroacetic anhydride
`total growth inhibition
`ala-3-(2-thienyl) or 2-amino-3-thiophen-2-yl
`propionic acid
`1,2,3,4-tetraisoquinoline-3-carboxilic acid
`undecanoic acid.
`
`N(CH2).N'(Me)-Arg
`
`MeOH
`N(Me.Ph),N'(Me)-Arg
`
`NMM
`MSt-OH
`NaI
`Oct-OH
`6-OHep-OH
`Oic
`N(CH N(CH), N(CH2))-Orn
`Plf-OH
`Phg
`(5R)-Ph-Pro
`Pip
`Pipe-OH
`SPS
`Tfa
`TFA
`TFAA
`TGI
`Thi
`
`Tic
`Und-OH
`
`0061 Amino acid symbols denote the L-configuration
`unless stated otherwise.
`0062) Examples of compounds of this invention include
`those of formula 1 wherein there is:
`0063 Glu or Lys instead of L-Orn in position 8:
`0064 Gly, Phe, Ala, Leu, D-Val, Pro, Gln, Orn, Thr or
`Glu instead of L-Val in position 11;
`0065 Val or D-Thr instead of L-Thr in position 12:
`0.066
`Gly, D-Ala, D-Leu, D-Phe, D-Pro, Val, D-Glu,
`D-Gln or D-Thr instead of D-Val in position 13;
`0067 hdh instead of L-Val in position 11 and/or D-Cha
`instead of D-Val in position 13;
`0068 Ala, Gly, Leu, Pro, Glu, Orn or Gln instead of
`L-Thr in position 12 and absence of amino acid in position
`13;
`0069 D-Ile or D-Val instead of D-allo-Ile in position 7:
`0070 D-Orn instead of L-Orn in position 8 and L-Pro
`instead of D-Pro in position 9;
`0071. D-Cys instead of D-allo-Ile in position 7 and L-Cys
`instead of L-Val in position 11; or
`0072 D-Cys or D-homo-Cys instead of D-allo-Ile in
`position 7 and D-Cys or D-homo-Cys instead of D-Val in
`position 10.
`0073. Icos, (c/t)-4-Me-cHexa, Und, (4R)-Mehex, (4RS)-
`MeHex, (4S)-Mehex, Oct, p-MeBza, Bza, p-CFBza, 3.5-
`dFPhAc, Pipe, p-CF,Cinn, p-CFPhAc, Pfh, 6-OHep, 6,6-
`dFHep, 4-GuBut, AM, AO or C(=N(CH)) instead of
`5-MeHex in position 14, and, optionally, Lys instead of
`L-Orn in position 8:
`
`0074) Pro, D-Pip, D-Tic or (5R)-Ph-Pro instead of D-Pro
`in position 9 and (4S)-MeHex instead of 5-MeHex in
`position 14'.
`N(Me.Ph).N'(Me)-Arg,
`0075 N(Me)N'(Me)-Arg,
`N(CH2)N(CH2)4-Arg,
`N(CH2)4N'(Me)-Arg,
`N(CHN(CH),N'(CH))-Orn, N(Me)-Lys, Orn(NTfa)
`or Orn(Biot) instead of L-Orn in position 8, and, optionally
`(4S)-MeHex instead of 5-MeHex in position 14 and, option
`ally, Thr(OTfa) instead of L-Thr in position 12:
`0.076 Thr(OTfa) instead of L-Thr in position 12 and
`(4S)-MeHex or Lit(OTfa) instead of 5-MeHex in position
`14:
`0.077 N-(Hep)-D-Val instead of D-Val in position 13
`and there is absence of 5-MeHex in position 14; or
`0078 absence of amino acids in position 11, 12 and 13,
`and, optionally, Mst instead of 5-MeHex in position 14.
`0079) Dha or E-Dhb instead of Z-Dhb in position 2:
`0080) D.L-Ser, Gly or Aib instead of Thr in position 2,
`being the analogues hydrogenated;
`0081. D-Val instead of L-Val in position 1:
`0082 Trp instead of L-Phe in position 3; or
`0.083 D-Thr or D-Ser instead of D-alo-Thr in position 6.
`0084 hCh, Phe (3.4-C), Phe (F), Phe (4-I), Phe
`(4-NO), Phe (4-F), Tyr (Me), Thi, Tic, Tyr, Oic, NMePhe,
`Phe (2-CI), Phe (3-Cl), Phe (4-Cl), Phe (3,4-F), NaI, Bip or
`Phg instead of L-Phe in position 3, and, optionally, (4S)-
`MeHex or p-CF,Cinn instead of 5-MeHex in position 14.
`0085 D-Val or D-Chainstead of D-alo-He in positions 5
`and 7 and, optionally, D-Chainstead of D-Val in position 4;
`O
`
`MPI EXHIBIT 1066 PAGE 8
`
`MPI EXHIBIT 1066 PAGE 8
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`D-Val instead of L-Val in position 1, D-Phe instead
`0.086
`of L-Phe in position 3, Val instead of D-Val in position 4.
`L-alo-Ile instead of D-alo-Ile in position 5, L-alo-Thr instead
`of D-alo-Thr in position 6, L-alo-Ile instead of D-alo-Ile in
`position 7, D-Orn instead of L-Orn in position 8, L-Pro
`instead of D-Pro in position 9, L-Val instead of D-Val in
`position 10, D-Val instead of L-Val in position 11, D-Thr
`instead of L-Thr in position 12 and L-Val instead of D-Val
`in position 13.
`0087 As appropriate, these suggested changes may be
`taken in combination.
`0088. The present invention also encompass the pharma
`ceutically acceptable salts, prodrugs, tautomers, and Sol
`vates, thereof.
`0089. The compounds of the present invention have
`asymmetric centers and therefore exist in different enantio
`meric and diastereomic forms. This invention relates to the
`use of all optical isomers and stereoisomers of the com
`pounds of the present invention, and mixtures thereof, and to
`all pharmaceutical compositions and methods of treatment
`that may employ or contain them.
`0090 The compounds of the invention may be in crys
`talline form either as free compounds or as Solvates (e.g.
`hydrates) and it is intended that both forms are within the
`scope of the present invention. Methods of solvation are
`generally known within the art.
`0.091 The present invention also includes the compounds
`of the present invention, and the pharmaceutically accept
`able salts thereof, wherein one or more hydrogen, carbon or
`other atoms are replaced by isotopes thereof. Such com
`pounds may be useful as research and diagnostic tools in
`metabolism pharmacokinetic studies and in binding assays.
`0092. As used herein, the compounds of this invention,
`including the compounds of formula 1, are defined to
`include pharmaceutically acceptable derivatives or prodrugs
`thereof A “pharmaceutically acceptable derivative or pro
`drug' means any pharmaceutically acceptable salt, ester, salt
`of an ester or other derivative of a compound of this
`invention that, upon administration to a recipient, is capable
`of providing (directly or indirectly) a compound of this
`invention or a metabolite or residue thereof. Particularly
`favoured derivatives and prodrugs are those that increase the
`bioavailability of the compounds of this invention when
`Such compounds are administered to a patient (e.g., by
`
`allowing an orally administered compound to be more
`readily absorbed into the blood), enhance delivery of the
`parent compound to a given biological compartment,
`increase solubility to allow administration by injection, alter
`metabolism or alter rate of excretion.
`0093 Salts of the compounds of the present invention
`may comprise acid addition salts derived from a nitrogen in
`the compound of formula 1 or 2. The therapeutic activity
`resides in the moiety derived from the compound of the
`invention as defined herein and the identity of the other
`component is of less importance although for therapeutic
`and prophylactic purposes it is, preferably, pharmaceutically
`acceptable to the patient. Examples of pharmaceutically
`acceptable acid addition salts include those derived from
`mineral acids, such as hydrochloric, hydrobromic, phospho
`ric, metaphosphoric, nitric and Sulphuric acids, and organic
`acids, Such as tartaric, acetic, trifluoroacetic, citric, malic,
`lactic, fumaric, benzoic, glycolic, gluconic, Succinic and
`methaneSulphonic and arylsulphonic, for example p-tolu
`enesulphonic, acids. Preferred salts include the trifluoroac
`etate salt and the hydrochloride salt.
`0094. The compounds of the present invention can be
`prepared according to the synthetic process described in WO
`0158934, or according to the improved process as described
`herein. Therefore also encompassed by the invention is a
`process to prepare a compound according to formula 1.
`0095 The key steps of the optimized process for a more
`economical and safe synthesis of kahalalide F and its
`analogues are: (i) partial incorporation of Fmoc-D-Val-OH
`onto the chlorotritylchloro-polystyrene resin for a initial
`loading of 0.5 mmol/g; (ii) use as coupling method of
`DIPCDI-HOBt, instead of HATU-DIPEA, for the sequential
`incorporation of the protected amino acids and aliphatic
`carboxylic acids; (iii) cyclization step with DIPCDI/HOBt/
`DIPEA in CHCl; these conditions avoids two side reac
`tion: epimerisation of the Val residue, which is involved in
`the activation, and trifluoroacetylation of the Phe or its
`replacement; (iv) use of sodium diethyl-dithiocarbamate
`after removing Alloc to avoid presence of Pd(0) in the final
`product.
`0096. The synthesis is preferably a solid phase synthetic
`process.
`0097. The preferred embodiment of the synthetic process
`of the present invention is best represented in the Scheme 1,
`which is directed to the formation of the target compounds.
`
`Scheme 1
`
`Fmoc-DVal-O-O as- CI-O
`
`|
`
`N
`
`C
`
`MPI EXHIBIT 1066 PAGE 9
`
`MPI EXHIBIT 1066 PAGE 9
`
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`

`US 2007/01 17743 A1
`
`9
`
`May 24, 2007
`
`-continued
`Fmoc-Dae-DaThr-Dale-DVal-O -O
`OH
`
`Fmoc-Dalle-Dahr-Dalle-DVal-O-O
`
`O-Wal-Alloc
`
`t
`
`MeHex-DVal-Thr- Wal-DVal-DPro-On-Dalle-DaThr-Dalle-DVal-O
`
`tBu
`
`Boc
`
`O-Wal-Alloc
`
`MeHex-DVal-Thr- Wal-DVal-DPro-On-Dalle-DaThr-Dalle-DVal-O
`
`tBu
`
`Boc
`
`O-Wal-ZDhb-Phe-Alloc
`
`MeHex-DVal-Thr-Val-DVal-DPro-On-Dalle-DaThr-Dalle-DVal-O
`
`tBu
`
`Boc
`
`O-Val-ZDhb-Phe-H
`
`MeHex-DVal-Thr-Wal-DVal-DPro-On-Dale-DaThr-Dale-DVal-OH
`
`tBu
`
`Boc
`
`O-Wal-ZDhb-Phe-H
`
`MeHex-DVal-Thr-Val-DVal-DPro-On-Dalle-DaThr
`
`Dale
`
`DVal
`
`tBu
`
`Boc
`
`O-Wal-ZDhb-Ph.
`
`MeHex-DVal-Thr-Val-DVal-DPro-On-Dalle-DaThr
`
`Dale
`
`DVal
`
`O-Wal-ZDhb-Phe
`
`0.098 As shown above in Scheme 1, the preferred process
`for the synthetic formation of analogues of kahalalide F is
`based in a Solid-phase approach, see for example Lloyd
`Williams, P. et al. Chemical Approaches to the Synthesis of
`Peptides and Proteins. CRC Press, Boca Raton (Fla.), 1997
`and followed with modifications of the method already
`described for the preparation kahalalide F and some of its
`analogues (WO 0158934).
`0099. The process of Scheme 1 comprises the sequential
`steps of
`0100 (a) incorporating an Fmoc-DVal-OH onto a chlo
`rotrityl chloro resin, forming an ester bond;
`0101 (b) elongating the peptidic chain with three amino
`acids Dalle, DaThr (free OH), Dale) using a Fmoc/tBu
`Strategy:
`
`0102 (c) incorporating Val(1) using an Alloc/tEu strat
`egy,
`(d) elongating the peptidic chain with the remain
`0.103
`ing amino acids and aliphatic carboxylic acids using a
`Fmoc/tBu strategy:
`0.104
`(e1) incorporating the dipeptide Alloc-Phe-ZDhb
`OH, which has been combined and dehydrated in solution;
`0105 (e2a) elongating the peptidic chain with two amino
`acids, preferably Thr and Phe. The OH of Thr is unprotected
`and the amino group of Phe, or its replacement, is protected
`with Fmoc or preferably with Alloc; in some cases if it is
`protected with Fmoc, this is removed and Alloc is introduced
`in Solid-phase;
`0106 (e2b) dehydrating in solid-phase to give the dide
`hydropeptide;
`
`MPI EXHIBIT 1066 PAGE 10
`
`MPI EXHIBIT 1066 PAGE 10
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`

`US 2007/01 17743 A1
`
`May 24, 2007
`
`0107 (f) removing the Alloc/Fmoc group of Phe, or of its
`replacement, while the peptide is still anchored to the solid
`Support;
`0108 (g) cleaving the side-chain protected peptide from
`the Solid Support;
`0109 (h) cyclizing the peptide in solution;
`0110 (i) removing TFA labile side chain protecting
`groups.
`() Further modifications of functional(s) group(s)
`0111
`in Solution phase.
`Therefore the process can be conducted as follows:
`0112 Fmoc-DVal-OH is incorporated preferably to a
`chlorotrityl-polystyrene resin, see Barlos, K. Gatos, D.;
`Schäfer, W. Angew. Chem. Int. Ed. Engl. 1991, 30, 590-593,
`in the presence of DI