(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT)
`
`(19) World Intellectual Property
`Organization
`International Bureau
`
`•
`
`111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111111
`
`( 43) International Publication Date
`10 February 2005 (10.02.2005)
`
`PCT
`
`(10) International Publication Number
`WO 2005/012327 A2
`
`(51) International Patent Classification7:
`A61K 31/7068, 31/7072, A61P 35/00
`
`C07H 19/10,
`
`(21) International Application Number:
`PCT /GB2004/003148
`
`(22) International Filing Date:
`
`20 July 2004 (20.07.2004)
`
`(25) Filing Language:
`
`(26) Publication Language:
`
`(30) Priority Data:
`0317009.9
`
`English
`
`English
`
`21 July 2003 (21.07.2003) GB
`
`(71) Applicant (for all designated States except US): UNI(cid:173)
`VERSITY COLLEGE CARDIFF CONSULTANTS
`LIMITED [GB/GB]; P.O. Box 497, 30-36 Newport Road,
`Cardiff CF24 ODE (GB).
`
`-iiiiiiiiiiii
`
`iiiiiiiiiiii
`
`(72) Inventor; and
`(75) Inventor/Applicant (for US only): MCGUIGAN,
`Christopher [GB/GB]; 2 Alfreda Road, Whitchurch,
`Cardiff CF4 2EH (GB).
`
`(74) Agents: HOWARD, Paul, Nicholas et al.; Carpmaels &
`Ransford, 43-45 Bloomsbury Square, London WC1A 2RA
`(GB).
`
`(81) Designated States (unless otherwise indicated, for every
`kind of national protection available): AE, AG, AL, AM,
`AT, AU, AZ, BA, BB, BG, BR, BW, BY, BZ, CA, CH, CN,
`CO, CR, CU, CZ, DE, DK, DM, DZ, EC, EE, EG, ES, Fl,
`GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE,
`KG, KP, KR, KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD,
`MG, MK, MN, MW, MX, MZ, NA, NI, NO, NZ, OM, PG,
`PH, PL, PT, RO, RU, SC, SD, SE, SG, SK, SL, SY, TJ, TM,
`TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, YU, ZA, ZM,
`zw.
`
`(84) Designated States (unless otherwise indicated, for every
`kind of regional protection available): ARIPO (BW, GH,
`GM, KE, LS, MW, MZ, NA, SD, SL, SZ, TZ, UG, ZM,
`ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, RU, TJ, TM),
`European (AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, Fl,
`FR, GB, GR, HU, IE, IT, LU, MC, NL, PL, PT, RO, SE, SI,
`SK, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ,
`GW, ML, MR, NE, SN, TD, TG).
`
`[Continued on next page}
`
`::: --------------------------------------------------------------------------------------
`
`---
`iiiiiiiiiiii ----iiiiiiiiiiii
`iiiiiiiiiiii ----
`
`(54) Title: CHEMICAL COMPOUNDS
`
`300 -
`
`225 -
`
`150 -
`
`75
`
`ns
`
`0.06!-lg/ml
`ns
`
`ns
`
`l I
`
`**
`*
`I I
`I
`10 1-LM
`CPF31
`
`5!-lM
`CPF31
`
`0.1 1-LM
`1!-lM
`CPF31
`CPF31
`D Tumour volume
`p=0.096 vs control; ** p=0.094 vs control
`
`I
`
`0.01 1-LM
`CPF31
`
`I
`
`0
`
`Control
`
`*
`
`t--(cid:173)
`M
`~
`M
`,..-.!
`0 .........
`ln
`0
`0
`M
`0 of, for example, each of deoxyuridine, cytarabine, gemcitabine and citidine may be substituted at the 5-position. The phosphorami(cid:173)
`> date moiety has attached to the P atom an aryl-0 moiety and an a-amino acid moiety. The a-amino acid moiety may correspond to
`
`(57) Abstract: Phosphoramidate derivatives of nucleotides and their use in the treatment of cancer are described. The base moieties
`
`~ or be derived from either a naturally occurring or a non-naturally occurring amino acid.
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`W 0 2005/0123 2 7 A2
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`lllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllllll
`
`Published:
`without international search report and to be republished
`upon receipt of that report
`
`For two-letter codes and other abbreviations, refer to the "Guid-
`ance Notes on Codes and Abbreviations" appearing at the begin-
`ning of each regular issue of the PCT Gazette.
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`Chemical Compounds
`
`The present invention relates to nucleotide derivatives and their use in the treatment of
`
`cancer.
`
`5
`
`Nucleoside analogues such as fluorodeoxyuridine (1), cytarabine (2) and gemcitabine (3)
`
`are well established as anticancer agents. They function as inhibitors of DNA synthesis
`
`after activation to their 5' -phosphate form.
`
`0
`HNJ!.yF
`0~)
`
`Hol ... o .. /
`
`10 ~ (1)
`
`The free bioactive phosphate forms do not in general represent useful drugs due to their
`
`poor membrane permeation. In an effort to circumvent this a number of phosphate pro(cid:173)
`
`drug approaches have been reported [Rosowsky et al, J. Med. Chern., 1982, 25, 171-8;
`
`15 Hong et al, J. Med. Chern., 1985,28, 171-8; Kodama et al, Jpn. J. Cancer Res., 1989, 80,
`
`679-85; Hong et al, 1979, 22, 1428-32; Ji et al, J. Med. Chern., 1990, 33, 2264-70; Jones et
`
`al, Nucleic Acids Res., 1989, 17, 7195-7201; Hunston et al, J. Med. Chern., 1984, 27, 440-
`
`4; Lorey et al, Nucleosides Nucleotides, 1997, 16, 1307-10; Farquhar et al, J. Med. Chern.,
`
`1983, 26, 1153-8; Shuto et al, Nucleosides Nucleotides, 1992, 11, 437-46; Le Bee et al,
`
`20 Tet. Letts., 1991,32, 6553-6; Phelps et al, J. Med. Chern., 1980,23, 1229-32].
`
`In general the phosphate prodrugs have biological properties and therapeutic activities that
`
`are similar to, or somewhat lower than, the parent nucleoside analogue.
`
`25 We have carried out extensive work in this area from an antiviral perspective, largely on
`
`dideoxy nucleosides, and have reported a phosphoramidate approach which has been
`
`widely adopted for the delivery ofbio-active phosphates of antiviral nucleosides.
`
`An example is the phosphoramidate (4) derived from anti-HIV d4T (5).
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`2
`
`0
`HN~
`0~ H00
`
`(5)
`
`0
`
`HNY
`0-J..NJ
`
`o
`-
`
`(4)
`
`II
`
`0 Me 0 0
`
`I
`
`I
`
`Me·O-C-CH-N-P-Q Ho
`F\
`\_/)
`
`We observed the effect of variations in the ester [McGuigan et al, AVCC, 1998, 9, 473-9],
`
`5 amino acid [McGuigan et al, Antiviral Res., 1997, 35, 195-204; A VCC, 2000, 11, 111-6],
`
`and aryl [Siddiqui et al, J. Med. Chern., 1999, 42, 393-9] regions of the phosphoramidate,
`
`as well as the effect of amino acid stereochemistry [McGuigan et al, A VCC, 1996, 7, 184-
`
`8], phosphate stereochemistry [Allender et al, Analytica Chim. Acta, 2001, 435, 107-13]
`
`and nucleoside [Balzarini et al, BBRC, 1996, 225, 363-9; McGuigan et al, BioOrg. Med,
`
`10 Chern. Lett., 1996, 6, 2369-62; McGuigan et al, Bioorg. Med. Chern. Lett., 2000, 10, 645-
`
`7].
`
`This work has lead to the optimal description of phenyl methoxyalaninyl phosphoramidate
`
`as the prototype pro-moiety for the intracellular delivery of bioactive nucleotides
`
`15
`
`[Balzarini et al, PNAS, 1996,93, 7295-9; McGuigan et al, J. Med. Chern., 1996,39, 1748-
`
`53].
`
`Lackey et al [Biochem Pharmacol., 2001, 61, 179-89] have reported the application of our
`
`phosphoramidate pro-drug method for antiviral nucleosides to the anti-herpetic agent
`
`20 bromovinyl-2'-deoxyuridine (BVDU) (6). In particular, they have found that the phenyl
`
`methoxyalaninyl phosphoramidate (7) has significant anti-cancer activity. This is in
`
`marked contrast to the parent (antiviral) nucleoside (6).
`
`0Me01;1
`
`I
`
`II
`
`II
`Me{) -C-CH-N-P -0
`He)
`;==\
`\__/
`
`0
`
`Br
`HNY
`OJ_.NjJ
`
`o
`
`(7)
`
`OH
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`3
`
`Limited SAR has been presented by this group, although in their patent applications
`
`[W00239952, EP1200455, CA2317505, US6339151, EP116797, AU2451601] they claim
`
`a series of general variations in the base, and phosphate regions. However, based on our
`
`5 prior art, the phenyl methoxyalaninyl phosphoramidate (7) would be anticipated to be
`
`amongst the most optimal of structures.
`
`Surprisingly,
`
`it has now been found
`
`that other derivatives of oxyamino acid(cid:173)
`
`phosphoramidate nucleoside analogues are significantly more potent in the treatment of
`
`1 0 cancer than the phenyl methoxyalaninyl phosphoramidate (7).
`
`According to a first aspect of the present invention there is provided a compound of
`
`formula I:
`
`Z'
`
`Hn~~
`~1
`0
`0
`II
`11-+R
`O
`N
`N-P-0~
`R-O-C
`I
`R"
`I
`Q
`H 0
`A/
`
`4
`
`z
`
`X
`y
`
`I
`
`.
`
`OH
`
`15
`
`wherein:
`
`R is selected from the group comprising alkyl, aryl and alkylaryl;
`
`R' and R" are, independently, selected from the group comprising H, alkyl and alkylaryl,
`
`orR' and R" together form an alkylene chain so as to provide, together with the C atom to
`
`20 which they are attached, a cyclic system;
`Q is selected from the group comprising -0- and -CH2-;
`X and Y are independently selected from the group comprising H, F, Cl, Br, I, OH and
`
`methyl ( -CH3);
`
`Ar is a monocyclic aromatic ring moiety or a fused bicyclic aromatic ring moiety, either of
`
`25 which ring moieties is carbocyclic or heterocyclic and is optionally substituted;
`
`Z is selected from the group comprising H, alkyl and halogen; and
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`n is 0 or 1,
`
`wherein
`
`PCT /GB2004/003148
`
`4
`
`when n is 0, Z' is -NH2 and a double bond exists between position 3 and position 4,
`
`and
`
`5
`
`when n is 1, Z' is ==0;
`
`or a phannaceutically acceptable derivative or metabolite of a compound of formula I;
`
`with the proviso that when n is 1, X andY are both H, R is methyl (-CH3), one ofR' and
`
`10 R" is H and one of R' and R" is methyl ( -CH3), then Ar is not phenyl ( -C6H5).
`
`By "a phannaceutically acceptable derivative" is meant any pharmaceutically acceptable
`
`salt, ester or salt of such ester or any other compound which upon administration to a
`
`recipient is capable of providing (directly or indirectly) a compound of formula (1).
`
`15
`
`Suitably, except where R is 2-Bu (-CH2-CH(CH3)2) and one of R' and R" isH and one of
`R' and R" is methyl ( -CH3), when n is 1 and X and Y are both H, then Ar is not
`
`unsubstituted phenyl ( -C6H5).
`
`20 By "phannaceutically acceptable metabolite" is meant a metabolite or residue of a
`
`Z'
`
`~ R"
`
`~
`
`R- O-C----:--rN-P-0~
`"
`I
`I
`Q
`R
`H OH
`
`4
`
`0~~ :r
`
`z
`
`(lij
`
`X
`y
`
`OH
`
`compound of formula (I) which gives rise in use to a compound of fonnula (II):
`
`wherein n, Q, R, R', R", X, Y, Z and Z' have the meanings described above and below for
`
`25
`
`fonnula I, and additionally R can be H, with the proviso that when n is 1, X and Y are both
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`H, R is methyl (-CH3), one ofR' and R" isH and one ofR' and R" is methyl (-CH3), then
`
`Z is not -CH=CHBr.
`
`5
`
`Suitably, with respect to compounds of formula II, when n is 1 and Z either is or is not -
`
`5 CH=CHBr, the moiety ROCOCR'R"NH- corresponds neither to alanine (ie as above, R is
`
`not methyl (-CH3), one ofR' and R" is not Hand one ofR' and R" is not methyl (-CH3))
`nor to tryptophan (ie a-amino-~-indolylpropionic acid).
`
`More suitably with respect to compounds of formula II, when n is 1 and Z either is or is not
`
`10
`
`-CH=CHBr, the moiety ROCOR'R"NH is neither derived from nor corresponds to any
`
`naturally ocurring amino acid.
`
`Even more suitably, with respect to compounds of formula II, when n is 1 or 0, the moiety
`
`ROCOCR'R''NH- does not correspond to alanine (ie R is not methyl (-CH3), one ofR' and
`
`15 R" is is not Hand one ofR' and R" is not methyl (-CH3)), does not preferably correspond
`
`to tryptophan, and even more preferably the said moiety does not correspond to any
`
`naturally ocurring amino acid.
`
`Most preferably the moiety ROCOCR'R"NH- in compounds of formula II corresponds to a
`
`20 non-naturally occurring amino acid.
`
`Reference in the present specification to an alkyl group means a branched or unbranched,
`
`cyclic or acyclic, saturated or unsaturated (e.g. alkenyl or alkynyl) hydrocarbyl radical.
`
`Where cyclic, the alkylene group is preferably C3 to C12, more preferably Cs to C10, more
`
`25 preferably C5 to C7. Where acyclic, the alkyl group is preferably C 1 to C1 6, more
`preferably cl to c6.
`
`Reference in the present specification to an aryl group means an aromatic group containing
`
`5 to 14 ring atoms, for example phenyl or naphthyl. The aromatic group may be a
`
`30 heteroaromatic group containing one, two, three or four, preferably one, heteroatoms
`
`selected, independently, from the group consisting of 0, N and S. Examples of such
`
`heteroaromatic groups include pyridyl, pyrrolyl, furanyl and thiophenyl. Preferably, the
`
`aryl group comprises phenyl or substituted phenyl.
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`6
`
`The alkyl and aryl groups may be substituted or unsubstituted. Where substituted, there
`
`will generally be one to three substituents present, preferably one substituent. Substituents
`
`may include halogen atoms, by which is meant F, Cl, Br and I atoms, and halomethyl
`
`5 groups such as CF3 and CCh; oxygen containing groups such as oxo, hydroxy, carboxy,
`
`carboxyC1-16alkyl, alkoxy, alkoyl, alkoyloxy, aryloxy, aryloyl and aryloyloxy; nitrogen
`
`containing groups such as amino, CJ.6alkylamino, diC1_6alkylamino, cyano, azide and nitro;
`sulphur containing groups such as thiol, C1.6alkylthiol, sulphonyl and sulphoxide;
`heterocyclic groups which may themselves be substituted; alkyl groups as defined above,
`
`10 which may themselves be substituted; and aryl groups as defined above, which may
`
`themselves be substituted, such as phenyl and substituted phenyl. Substituents on said
`
`heterocyclic, alkyl and aryl groups are as defined immediately above.
`
`Reference in the present specification to alkoxy and aryloxy groups means, respectively,
`
`15 alkyl-0- (for example where alkyl is C1 to C16, preferably C1 to C6) and aryl-0- (for
`example where aryl is a 5 to 14 membered aromatic mono- or bifused ring moiety,
`
`optionally containing 1, 2, 3 or 4 heteroatoms selected, independently, from 0, S and N,
`
`preferably aryl is phenyl).
`
`20 Reference in the. present specification to alkoyl and aryloyl groups means, respectively,
`
`alkyl-CO- (for example where alkyl is C1 to C16, preferably C1 to C6) and aryl-CO- (for
`example where aryl is a 5 to 14 membered aromatic mono or bifused ring moiety,
`
`optionally containing 1, 2, 3 or 4 heteroatoms selected, independently, from 0, S and N,
`
`preferably aryl is phenyl).
`
`25
`
`Reference in the present specification to alkoyloxy and aryloyloxy means, respectively,
`
`alkyl-CO-O (for example where alkyl is C1 to C16, preferably C1 to C6) and aryl-CO-O (for
`example where aryl is a 5 to 14 membered mono- or bifused aromatic ring system,
`
`optionally containing I, 2, 3 or 4 heteroatoms selected, independently, from 0, S and N,
`
`30 preferably aryl is phenyl).
`
`Reference in the present specification to heterocyclic groups means groups containing one
`
`or more, pyrrolyl, imidazolyl, pyraziolyl, thiazolyl, isothiazolyl, oxazolyl, pyrrolidinyl,
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`7
`
`pyrrolinyl,
`
`imidazolidinyl,
`
`imidazolinyl, pyrazolidinyl,
`
`tetrahydrofuranyl, pyranyl,
`
`pyronly, pyridyl, pyrazinyl, pyridazinyl, piperidyl, piperazinyl, morpholinyl, thionaphthyl,
`
`benzofuranyl, isobenzofuryl, indolyl, oxyindolyl, isoindolyl, indazolyl, indolinyl, 7-
`
`azaindolyl, isoindazolyl, benzopyranyl, coumarinyl, isocoumarinyl, quinolyl, isoquinolyl,
`
`5 naphthridinyl, cinnolinyl, quinazolinyl, pyridopyridyl, benzoxazinyl, quinoxadinyl,
`
`chromenyl, chromanyl, isochromanyl and carbolinyl.
`
`The group Ar comprises a substituted or unsubstituted aryl group, wherein the term "aryl
`
`group" and the possible substitution of said group is as defined herein. Preferably, Ar is a
`
`10 substituted or unsubstituted phenyl group. Particularly preferred substituents are electron
`
`withdrawing groups such as halogen (preferably chlorine or fluorine), trihalomethyl
`
`(preferably trifluoromethyl), cyano and nitro groups. For example, Ar can be phenyl, 3,5-
`
`dichloro-phenyl, n-trifluoromethyl-phenyl, n-cyano-phenyl, or n-nitro-phenyl. When Ar is
`
`a heteroaromatic group, preferably it is optionally substituted pyridyl.
`
`15
`
`Suitably, R is a Ct-16 primary or secondary alkyl group, a C5_7 carbocyclic aryl group or a
`CI-6alky1Cs.uaryl group. More suitably, R is a Ct.Jo alkyl group, a phenyl group or CI-3
`
`alkylCs-7 aryl group. Preferably R is unsubstituted.
`
`20 Preferably, R is methyl ( -CH3), ethyl ( -CzHs), n- or i- propyl ( -C3H7), n- or i- butyl ( -C4H9)
`or benzyl ( -CHzC6Hs). Most preferably, R is benzyl. Particularly, R is preferably benzyl
`
`when one ofR' and R" isH and one ofR' and R" is methyl (-CH3), especially when Ar is
`
`unsubstituted phenyl, n is 0 and each ofX andY is F.
`
`25 Suitably, R' and R" are each independently selected from the group comprising H, C1.6
`primary, secondary or tertiary alkyl, Ct-3alkylC5•7aryl, or, when together they form an
`alkylene chain, they provide, together the C atom to which they are attached, a C3•8
`carbocyclic aliphatic ring.
`
`30 Preferably, R' and R" are the same and are alkyl, more prefearbly they are both methyl,
`
`ethyl or n- or i- propyl.
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`
`Alternatively, preferably, R' and R" are, independently, H, methyl ( -CH3), secondary butyl
`
`(-CH2-CH-(CH3)2), benzyl (-CH2C6H5), or, together with the C atom to which they are
`attached, provide a Cs-6 ring.
`
`5 Preferred compounds include those where R' and R" are both methyl, one of R' and R" is
`
`Hand one ofR' and R" is methyl, and R' and R", together with the C atom to which they
`
`are attached, provide a pentyl ring.
`
`When R' and R" are different, the C atom to which they are attached is chiral. The present
`
`10 compounds can be L or D or a mixture of stereoiosomers. Preferably they are L.
`
`It will be appreciated that the moiety -0-C(O)-CR'R"-NH- corresponds to a carboxy(cid:173)
`
`protected a-amino acid. R' and R" can thus correspond to the side chains of a naturally
`
`occurring amino acid.
`
`15
`
`For example, when one of R' and R" is H and one of R' and R" is Me or PhCH2, the
`
`moiety corresponds to alanine or phenylalanine, respectively.
`
`Preferably, the stereochemistry at the asymmetric centre -CR'R" corresponds to an 1-
`20 amino acid. The stereochemistry at the asymmetric centre -CR'R" can, however,
`
`correspond to a D-amino acid. Alternatively, mixtures of of compounds can be employed
`having asymmetric centres corresponding to 1 and D amino acids.
`
`In the present specification by "naturally occurring amino acid" we mean Alanine,
`
`25 Arginine, Asparagine, Aspartic Acid, Cysteine, Cystine, Glycine, Glutamic Acid,
`
`Glutamine, Histidine, Hydroxylysine, Hydroxyproline, Isoleucine, Leucine, Lysine,
`
`Methionine, Phenylalanine, Proline, Serine, Threonine, Tryptophan, Tyrosine and Valine.
`
`The present invention
`
`is not, however,
`
`limited
`
`to compounds having a moiety
`
`30 corresponding to a naturally occurring amino acid. The present invention specifically
`
`includes compounds having a moiety which corresponds to a non-naturally occurring
`
`amino acid, such as, for example, those where R'=R"=alkyl, or, where together with the C
`
`atom to which they are attached, R' and R" provide a cyclic moiety. Preferably with
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`respect to the compound of formula I, the moiety ROCOCR'R''NH- corresponds to or is
`
`derived from a non-naturally occurring amino acid.
`
`9
`
`With respect to compounds of formula I when n is 1, the moiety ROCOCR'R"NH-
`
`5 preferably neither corresponds to nor is derived from alanine, more preferably neither
`
`corresponds to nor is derived from either of alanine or tryptophan, even more preferably
`
`neither corresponds to nor is derived from any naturally occurring amino acid.
`
`With respect to compounds of formula I when n is 0, the moiety ROCOCR'R"NH-
`
`1 0 preferably neither corresponds to nor is derived from alanine, more preferably neither
`
`corresponds to nor is derived from either of alanine or trytophan, even more preferably
`
`neither corresponds to nor is derived from any naturally occurring amino acid.
`
`Preferably Q is 0.
`
`15
`
`Preferably, X andY are, independently, selected from the group comprising F, Hand OH.
`
`When n is 1, preferably each ofX andY is H.
`
`20 When n is 0, preferably each of X andY is F, or X is OH and Y isH, or X is H andY is
`
`OH.
`
`When Z is F, Q is 0, n is 1 and X and Y are each H, the base moiety of the compound of
`
`formula I corresponds to that offluorodeoxyuridine i.e. compound (1) above.
`
`25
`
`When Z is H, Q is 0, n is 0 and X is OH and Y is H, the base moiety of the compound of
`
`formula I corresponds to that of cytarabine i.e. compound (2) above.
`
`When Z isH, Q is 0, n is 0 and X andY are each F, the base moiety of the compound of
`
`30
`
`formula I corresponds to that of gemcitabine i.e. compound (3) above.
`
`When Z is H, Q is 0, n is 0 and X is H and Y is OH, the base moiety of the compound of
`
`formula I corresponds to that of cytidine.
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`10
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`Compounds of formula I wherein n is 0 and X and Y are F are preferred. Particularly
`preferred are compounds of formula I wherein n is 0, X andY are F, Q is 0 and Z isH,
`
`corresponding to phosphoramidated gemcitabine.
`
`5
`
`Also preferred are compounds of formula I wherein n is 0 and X is OH and Y is H.
`
`Particularly preferred are compounds of formula I wherein n is 0, X is OH, Y is H, Q is 0
`
`and Z is H, corresponding to phosphoramidated cytarabine.
`
`10 Also preferred are compounds of formula I wherein n is 0 and X is H and Y is OH.
`
`Particularly preferred are compounds of formula I wherein n is 0, X is H, Y is OH, Q is 0
`
`andZ is H, corresponding to phosphoramidated cytidine.
`
`Suitably, Ar is a 5 to 14 membered aromatic ring moiety. The one or two rings may
`
`15
`
`include 1, 2, 3 or 4 heteroatoms, preferably 1, selected, independently, from 0, Sand N.
`
`Preferably, Ar is a carbomonocyclic aromatic ring moiety. More preferably, Ar is a C6
`monocyclic aromatic ring moiety, ie is optionally substituted phenyl.
`
`20 One, two, three or four substituents, which may be the same or different, may be present on
`
`Ar and are selected from the group comprising halogen, which may -F, -Cl, -Br or -1; -
`
`NOz; -NHz; optionally substituted -C1-3alkyl; optionally substituted -C 1_3alkoxy, preferably
`methoxy (-OCH3); optionally substituted -SC1_3alkyl; -CN; optionally substituted -COC1_
`3alkyl; and optionally substituted -C02C 1_3alkyl. The optional substitutents are one or
`25 more up to six, preferably three, members selected from the group comprising halogen
`
`which may be F, Cl, Brand I and N02. Preferred substituents on Ar include F, Cl, CF3,
`
`and NOz.
`
`The substituents may be at any position on the ring moiety. Where the ring moiety is C6 ie
`
`30 phenyl, a single substituent at the 2 (ortho) or 4 (para) position is preferred. Where Ar is
`
`phenyl, a single substituent at the 4 position is more preferred.
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`Preferably, Ar is an optionally substituted phenyl moiety. More preferably, Ar is selected
`
`form the group comprising: Ph-, pCF3C6H4-, pFC6H4-, pN02C6H4-, pC1C6H4- and
`
`oClC6H4-.
`
`5 Suitably, Z is selected from the group comprising H, C1-6 alkyl, substituted C1.6 alkyl, C1.6
`alkenyl, substituted cl-6 alkenyl, cl-6 alkynyl, substituted cl-6 alkynyl and halogen, where
`
`halogen is F, Cl, Br or I. Substituents that may be present on the alkenyl or alkynyl moiety
`
`are selected from the group comprising F, Cl, Br, I, and -C02Me. One, two or three
`
`substituents may be present. The alkenyl and alkynyl groups may contain one or more sites
`
`10 ofunsaturation.
`
`Where Z is substituted alkenyl or alkynyl, the substituent is preferably on the terminal C
`
`atom.
`
`15 Preferably Z is selected from the group comprising H, F, optionally substituted C1.6alkyl
`particularly Me (-CH3), optionally substituted C 1.6alkenyl and optionally substituted C1•
`6alkynyl, the optional substituents being as recited immediately above.
`
`When n is 1, Z' is 0, Q is 0 and X and Y are each H, preferably Z is a substituted C2
`20 alkenyl (i.e. ethenyl or vinyl) moiety (-CH=CH-); more preferably, Z is bromovinyl ((cid:173)
`
`CH=CHBr) or methylpropenoate (-CH=CHC02Me); and most preferably, Z is -
`
`CH=CHBr.
`
`With respect to compounds of formula II, preferably when n is 1 and X and Y are both H,
`
`25
`
`then Z is not F.
`
`With respect to compounds of formula II, when n is 0, preferably X is not H and Y is not
`
`OH, more preferably X is OH andY isH or X andY are both F.
`
`30 With respect to compounds of formula II, when n is 0, X is OH and Y is H, preferably
`
`neither R' nor R" is phenylmethyl (ie benzyl) or 3-methylindolyl (ie 3-CH2indolyl).
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`Surprisingly, modifYing the ester moiety in compound (7) has been found to show a
`
`marked increase in potency with respect to cancer cell lines. A preferred compound
`
`embodying the present invention is the benzyl ester (8). It has surprisingly been found that
`
`the benzyl ester (8) is very significantly more potent against several cancer cell lines than
`
`5
`
`the methyl ester (7):
`
`Compound (8) inhibits the growth of colon cancer cell line HT115 by 50% at 1.4 f.1M,
`
`10 whilst (7) requires a concentration of 244 f.1M; (8) is thus 174 times more potent.
`
`Compound (8) is also 8 times more potent than (7) versus prostate cancer cell line PC-3
`
`(19 flM vs. 155 f.1M).
`
`The degree of potency enhancement for (8) vs. (7) is surprising based on the prior art.
`
`15 Thus, comparing the equivalent phosphoramidates of d4T reveals a ca 4-fold potency boost
`
`of (1 0) over (9) [McGuigan et al, A VCC, 1998, 9, 473-9].
`
`0 HNY
`. O~NJ
`Me-o-~-c~:-~-0-.
`/
`H 6
`'\::)
`d -
`
`(9)
`
`20 This would imply that the benzyl phosphoramidate motif in (1 0) is ca 4-fold more efficient
`
`at the intracellular delivery of the bio-active free phosphate forms of d4T than is the
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`methyl ester (9). A person skilled in the art would anticipate a similar degree of
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`enhancement for the benzyl phosphoramidate ofBVDU (8) over the methyl ester (7) whilst
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`we observed an almost 200-fold enhancement for colon cancer as noted above.
`
`13
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`Surprising efficacy of modifications in the amino acid and aryl moieties of the BVDU
`
`5 phosporamidate has also been found in compounds embodying the present invention.
`
`Thus, compound (11) has simultaneous modification in these two regions, being the p(cid:173)
`
`trifluoromethylphenyl benzyl [ a,a-dimethylglycinyl ]phosphoramidate.
`
`10
`
`Compound II shows high potency against a range of cancer cell types and is significantly
`
`and surprisingly more potent than (7). Thus, for breast cancer (11) is 60-fold more active
`
`(l.31J.M vs 79 !J.M), and for prostate cancer (11) is 254-fold more potent (0.61 !J.M vs. 155
`
`15
`
`!J.M). Against colon cancer, (11) is 35-fold more potent (7 !J.M vs 244 !J.M). Again, the
`
`degree of enhancement of the analogue (11) vs. (7) is surprising based on prior art. Thus,
`
`comparing (12) [dimethyl glycine modification] and (13) [p-CF3phenyl modification] to
`
`(9) shows no significant difference in potency.
`
`O~N
`
`0 HNY
`~ 'C/
`d -
`
`OMe
`Me-o-g7-1-P-O
`Me H 6
`
`(12)
`
`20
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`Thus 50% effective doses vs HIV-1 for (9), (12) and (13) are: 0.075, 0.29, and 0.01 J.lM
`
`respectively; within experimental error, (12) and (13) are identical in potency to (9). Thus a
`
`person skilled in the art would have predicted that (11) would show little enhancement
`
`over (7) as opposed to the 35 to 254-fold enhancements noted above.
`
`5
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`10
`
`Thus, compounds embodying the present invention and having variations in one or more of
`
`the ester (R), amino acid (R', R") and aryl (Ar) region of the phosphoramidate structure
`
`compared to phenyl methoxyalaninyl phosphoramidate can give surprising and substantial
`
`potency boosts of pro-tides derived from BVDU against a range of cancer cell types.
`
`According to a further aspect of the present invention there is provided a compound having
`
`formula I according to the present invention for use in a method of treatment, preferably in
`
`the prophylaxis or treatment of cancer.
`
`15 According to a further aspect of the present invention there is provided a method of
`
`phrophylaxis or treatment of cancer comprising administration to a patient in need of such
`
`treatment an effective dose of a compound having formula I according to the present
`
`invention.
`
`20 According to a further aspect of the present invention there is provided use of a compound
`
`having formula I of the present invention in the manufacture of a medicament for use in the
`
`treatment or prophlylaxis of cancer.
`
`According to a further aspect of the present invention there is provided a pharmaceutical
`
`25 composition comprising a compound having formula I of the present invention in
`
`combination with a pharmaceutically acceptable excipient, carrier or diluent.
`
`According to a further aspect of the present invention there is provided a method of
`
`preparing a pharmaceutical composition comprising the step of combining a compound
`
`30 having formula I of the present invention with a pharmaceutically acceptable excipient,
`
`carrier or diluent.
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`15
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`The present invention is particularly applicable for the treatment of a patient having breast
`
`cancer, colon cancer or prostate cancer. Examples of such cancers include breast MDA
`
`MB231, colon HT115 and prostate PC-3.
`
`5 The compound having formula I or pharmaceutical composition according to the present
`
`invention can be administered to a patient, which may be human or animal, by any suitable
`
`means.
`
`The medicaments employed in the present invention can be administered by oral or
`
`10 parenteral routes, including intravenous, intramuscular, intraperitoneal, subcutaneous,
`
`transdermal, airway (aerosol), rectal, vaginal and topical (including buccal and sublingual)
`
`administration.
`
`For oral administration, the compounds of the invention will generally be provided in the
`
`15
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`form of tablets or capsules, as a powder or granules, or as an aqueous solution or
`
`suspension.
`
`Tablets for oral use may include the active ingredient mixed with pharmaceutically
`
`acceptable excipients such as
`
`inert diluents, disintegrating agents, binding agents,
`
`20
`
`lubricating agents, sweetening agents,
`
`flavouring agents, colouring agents and
`
`preservatives. Suitable inert diluents include sodium and calcium carbonate, sodium and
`
`calcium phosphate, and
`
`lactose, while cornstarch and alginic acid are suitable
`
`disintegrating agents. Binding agents may include starch and gelatin, while the lubricating
`
`agent, if present, will generally be magnesium stearate, stearic acid or talc. If desired, the
`
`25
`
`tablets may be coated with a material such as glyceryl monostearate or glyceryl distearate,
`
`to delay absorption in the gastrointestinal tract.
`
`Capsules for oral use include hard gelatin capsules in which the active ingredient is mixed
`
`with a solid diluent, and soft gelatin capsules wherein the active ingredient is mixed with
`
`30 water or an oil such as peanut oil, liquid paraffin or olive oil.
`
`Formulations for rectal administration may be presented as a suppository with a suitable
`
`base comprising for example cocoa butter or a salicylate.
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`Formulations suitable for vaginal administration may be presented as pessaries, tampons,
`
`creams, gels, pastes, foams or spray formulations containing in addition to the active
`
`ingredient such carriers as are known in the art to be appropriate.
`
`5
`
`For intramuscular, intraperitoneal, subcutaneous and intravenous use, the compounds of
`
`the invention will generally be provided in sterile aqueous solutions or suspensions,
`
`buffered to an appropriate pH and isotonicity. Suitable aqueous vehicles include Ringer's
`
`solution and isotonic sodium chloride. Aqueous suspensions according to the invention
`
`10 may include suspending agents such as cellulose derivatives, sodium alginate, polyvinyl(cid:173)
`
`pyrrolidone and gum tragacanth, and a wetting agent such as lecithin.
`
`Suitable
`
`preservatives for aqueous suspensions include ethyl and n-propyl p-hydroxybenzoate.
`
`The compounds of the invention may also be presented as liposome formulations.
`
`15
`
`In general a suitable dose will be in the range of 0.1 to 300 mg per kilogram body weight
`
`of the recipient per day. A preferred lower dose is 0.5 mg per kilogrm body weight of