ZA200609498B - Sulfonylethyl phosphorodiamidates for use in the treatment of cancer - Google Patents

Description

Sulfonylethyl phosphorodiamidates

Technical Field

[0001] This invention relates to sulfonylethyl and thioethyl phosphorodiamidates, formulations containing them, their pharmaceutical use, and their preparation and intermediates in their preparation. :

Background Ast

[0002] US Patent No. 5,556,942 fand PCT Publication No. WO 95/09863] discloses compounds of the formula vA

CY

© st [ RAR ; L

R Rr? rR? and their amides, esters, and salts, where:

L is an electron withdrawing leaving group; $* is ~S(=0)-, -S(=0)-, -S(=NH)-, -S(=O)(=NH)-, -8*(C;-C; alkyl)-, -Se(=0)-, -Se(=0) -Se(=NH)-, or ~Se(=0)(=NH)-, or is -O-C(=O)-, or -HN-C(=0)~; each R', R* and R’ is independently H or a non-interfering substituent; nis0,1o0t2;

Y is selected from the group consisting of

HNCH(CH)~ HOOC(CH)mGH— HaNCH(CH)mCONHCH—

COOH NH, COOH

HOOC(CHz)n CHCONHCH;—

NH, where mis 1 or 2; and

AA_ is an amino acid linked through a peptide bond to the remainder of the compound.

[0003] The compounds are stated to be useful drugs for the selective treatment of target tissues which contain compatible GST isoenzymes, and simultaneously elevate the levels of

GM progenitor cells in bone marrow. Disclosed embodiments for L include those that generate a drug that is cytotoxic to unwanted cells, including the phosphoramidate and phosphorodiamidate mustaxds.

[0004] One of the compounds has the formula o SS

N—P—N of J he 0 0 ose ~ Ho LA COOH

NH, H 0 A Itis referred to in the patent as TER 286 and named as y-glutamyl-a-amino-B-((2-ethyl-

N,N,N. N-tetra(2-chloro)ethylphosphoramidate)sulfony) propionyl-(R)-(-)-phenylglycine. This compound, later referred to as TLIK286, has the CAS name L-y-glutamyl- 3-[[2-([bis[bis (2-chloroethylamino]phosphinylloxylethyljsulfonyl]-L-alanyl-2-phenyl- (2R)-glycine. As the neutral compound, its proposed Intemational Nonproprietary Name is canfosfamide; and as its hydrochloride acid addition salt, its United States Adopted Name is canfosfamide hydrochloride. Canfosfamide and its salts are anticancer compounds that are activated by the actions of GST P1-1, and by GST A1-1, to release the cytotoxic phosphorodiamidate mustard moiety.

[0005] Ir vitro, canfosfamide has been shown to be more potent in the M6709 human colon carcinoma cell line selected for resistance to doxorubicin and the MCF-7 human breast carcinoma cell line selected for resistance to cyclophosphamide, both of which overexpress

GST P1-1, over theit parental cell lines; and in murine xenografts of M7609 engineered to have high, medium, and low levels of GST P1-1, the potency of canfosfamide hydrochloride was positively correlated with the level of GST P1-1 (Morgan et al., Cancer Res., 58:2568 (1998).

[0006] Canfosfamide hydrochloride is currently being evaluated in multiple clinical trials for the treatment of ovarian, breast, non-small cell lung, and colorectal cancers. It has demonstrated significant single agent antitumor activity and improvement in survival in patients with non-small cell lung cancer and ovarian cancer, and single agent antitumor activity in colorectal and breast cancer. Evidence from i viz cell culture and tumor biopsies indicates _2_

that canfosfamide is non-cross-esistant to platinum, paclitaxel, and doxorubicin (Rosario et al, Mol. Pharmacol, 58:167 (2000), and also to gemcitabine. Patients treated with canfosfamide hydrochloride show a very low incidence of clinically significant hematological toxicity.

[0007] PCT Publication No. WO 95/09865 also discloses intermediates that are compounds of the formula re TRA: “ow © s/ rR\ee © s«[ RE\R® st [ RR 1 t 1 L L

R Rr? R3 R R2 R3 R' R2 R3 and their amides, csters, and salts, where:

L is an electron withdrawing leaving group;

Sis Sor Se;

S$* is =S(=0)-, -S(=0),-, -S(=NH)-, -S(=0)(=NH)-, -§*(C,-C, alkyl)-, -Se(=0)-, -Se(=O)y, -Se(=NH)-, ot —Se(=0)(=NH)-, ot is -O-C(=0)-, or -HN-C(=0)~; each R!, R* and R® is independently H or a non-interfeting substituent; nis0,10r2; :

YY is selected from the group consisting of

HNCH(CH)= HOOG(CH)nCH— HNGH(CHo)mCONHCHZ—

COOH NH, COOH

HOOC(CHp)nGHCONHCH; —

NH, where m is 1 or 2; and

AA, is an amino acid linked through a peptide bond to the remainder of the compound.

[0008] US Patent No. 6,506,739 [ and PCT Publication No. WO 01/83496] discloses compounds of the formula 0] pv

RQ \0-F~N

N hy

JT

X X where:

X is a halogen atom;

Qis O,S, or NH; and

R is hydrogen, optionally substituted lower alkyl, optionally substituted aryl, or optionally substituted heteroaryl, ot is RCO-, RNHCO-, R'SO;-, or R'NHSO,- whete R' is hydrogen, optionally substituted lower alkyl, optionally substituted aryl, ox optionally substituted heteroaryl; or R-Q together is chlorine; and their salts. .

[0009] The compounds ate stated to be antitumor agents.

[0010] It would be desirable to develop other anticancer drugs having an efficacy and safety as good or better than canfosfamide and other compounds of US Patent No. 5,556,942.

Disclosure of the invention

[0011] In a first aspect, this invention is compounds of formula A, B, and C:

R © R

NPN R, 2 R

R oo R N-E—N,

Ro § R R¢F

Nr ’

R o R SO. ‘ Re? SO, 0,8 2 0

R172 Ie R 9 R

Np NEN,

R R

K § R ©

A B Cc where: each R is independently hydrogen, C, ¢ alkyl, or -CH,CH,X, where each X is independently Cl,

Bt, C,, alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, ; alkyl, halo-C, , alkyl,

C, , alkyloxy, or halo-C, , alkyloxy, provided that at least two R’s in each phosphorodiamidate group are “CH,CH,X;

R' is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, ot optionally substituted heteroaralkyl; and

Ris optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally substituted arenediyl, optionally substituted atenedialkyl, optionally substituted heteroarenediyl, or optionally substituted heteroarenedialkyl, and their salts.

[0012] In a second aspect, this invention is pharmaceutical compositions comprising a compound of the first aspect of this invention.

[0013] In a third aspect, this invention is the use of compounds of the first aspect of this invention in the manufacture of medicaments to treat cancer; and methods of treating cancer by the administration of a compound of the first aspect of this invention or a pharmaceutical composition of the second aspect of this invention; alone or in combination with other anticancer therapies.

[0014] In a fourth aspect, this invention is compounds of formula BB and CC:

R 9 R

NP JAY i i

R R HTN

J Ro R

A g . _R? S g 7 R OO R

ANN N—p—N

R R

RO R ©

BB . CC where: each R is independently hydrogen, C, ¢ alkyl, or -CH,CH,X, where each X is independently Cl,

Br, C, , alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, ; alkyl, halo-C, ; alkyl,

C,; alkyloxy, ot halo-C, ; alkyloxy, provided that at least two R’s in each phosphorodiamidate group ate ~-CH,CH,X;

R! is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; and

R? is optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally substituted atenediyl, optionally substituted arenedialkyl, optionally substituted heteroarenediyl, or optionally substituted heteroarenedialkyl, and their salts.

[0015] In a fifth aspect, this invention is phatmacentical compositions comprising a compound of the fourth aspect of this invention.

[0016] In a sixth aspect, this invention is the use of compounds of the fourth aspect of this invention in the manufacture of medicaments to treat cancer; and methods of treating cancer by the administration of a compound of the fourth aspect of this invention or a pharmaceutical composition of the fifth aspect of this invention; alone or in combination with other anticancer therapies.

[0017] In a seventh aspect, this invention is methods of preparing compounds of the first and fourth aspects of this invention. Compounds of the first aspect of this invention are conveniently prepared from compounds of the fourth aspect of this invention.

Modes for carrying out the invention

[0018] Definitions

[0019] “Alkyl” means a monovalent group derived from a saturated or unsaturated (but not aromatically unsaturated) C,-C,, hydrocarbon that may be linear, branched, or cyclic by removal of one hydrogen atom from a carbon atom. Examples are methyl, ethyl, propyl, 1-ptopenyl, isopropyl, butyl, isobutyl, sec-butyl, zer-butyl, pentyl, hexyl, cyclopentyl, cyclopenten-1-yl, cyclopropylmethyl, cyclohexyl, and cyclohexylmethyl. Saturated alkyls (including cycloalkyls) and C,-C, alkyls are exemplary. Note that the definition of “alkyl” in this application is broader than the conventional definition and includes groups more commonly referred to as “cycloalkyl”, “cycloalkylalkyl”, “alkenyl”, and “alkynyl”. “Alkanediyl” means a divalent group derived from an “alkyl” (as that term is defined here) by removal of a second hydrogen atom from the same carbon atom or another carbon atom, preferably from another carbon atom. Examples are 1,2-ethanediyl, but-2-en-1,4-diyl, 1,5-pentanediyl, and 1,4-cyclohexanediyl.

[0020] A “substituted alkyl” is an alkyl substituted with up to three halogen atoms and/or up to three substituents selected from -CN, -NO,, ~OR, -SR, ~COR, —OC(O)R, —~C(O)OR, —NR, -NR,'X, -PR,, -PR,"X, -SO,0R, -OSO.R, -SO,;NR,, 2NRSO,R, -CONR,, -NRCOR, and -NRC(O)OR, whete each R is, independently, hydrogen, optionally R'-substituted alkyl, optionally R'-substituted heteroalkyl, optionally R'-substituted aryl, optionally R'-substituted heteroaryl, optionally R'-substituted aralkyl, or optionally R'-substituted heteroaralkyl and each

R'is, independently, 1 to 3 substituents selected from halo, -CN, -NO,, -OH, C, 5 alkyl, C,, alkyloxy, -SH, -NH,, or ~C(O)Oalkyl (preferably, 1 to 3 substituents selected from halo,

CN, -NO,, -OH, C, , alkyl, C, , alkyloxy, -SH, or -NH,), or two R groups form a 4- or 5-member optionally R'-substituted alkanediyl or optionally R'-substituted heteroalkanediyl, and X is a halogen. Thus, for example, substituted alkyl groups include such groups as trifluoromethyl, 3-chloropropyl, and 2-morpholinoethyl Substituted alkyl groups also include the residues (i.e. all except the thiol) of thiol-amino acids such as cysteine, homocysteine, and penicillamine, and their esters and amides formed by reaction at the carboxy group and amides or sulfonamides formed by reaction at the amine group, such as their N-protected forms and esters, where one substituent is -COOR and another substituent is “NH, or —NRC(O)OR. “Substituted alkanediyl” means alkanediyl substituted in the manner described above for an alkyl. Compounds of this invention also include compounds where any non-aromatic amine having 1 ot 2 hydrogen atoms present is protected by an amine-ptotecting group of the formula R*OC(O)- such as zert-butoxycatbonyl, benzyloxycarbonyl, fluorenylmethozycarbonyl, and other similar conventional carbamate-forming protecting groups. The term “substituted alkyl” specifically excludes dipeptides and higher peptides based on cysteine (ie. where the “substituted alkyl” is bonded to the sulfonylethyl phosphorodiamidate portion of the molecule through the cysteine sulfur atom) and their esters, amides, and ester/amides. “Elaborated” refers to the conversion of a reactive substituent to another typically more complex substituent, such as the convetsion of an amine to an amide or sulfonamide, a carboxy group to an ester or amide, a hydroxy to an ester, and convetsion of an amide or sulfonamide with one ot mote hydrogen atoms on the nitrogen to one where one or mote of those hydrogen atoms is teplaced by an optionally substituted alkyl, heteroalkyl, aryl, heteroaryl, aralkyl, or heteroaralkyl group. “Protected” has its conventional

Oy J meaning in organic synthesis, namely the temporary convetsion of a reactive substituent to 2 substituent that is non-reactive under the conditions of the reaction(s) proposed to be carried out; such as the protection of an amine as a carbamate as mentioned above.

[0021] “Heteroalkyl” means alkyl in which 1 to 3 of the carbon atoms are replaced by 0,S, or

NR (whete Ris H or C, , alkyl optionally substituted with halogen or hydroxy), including linear groups such as 3-oxapentyl; monocyclic rings containing 5 or 6 ring atoms such as 2-tetrahydrofuraayl, 2-pyrrolidinyl, 3-piperidinyl, 2-piperazinyl, 4-methyl-1-piperazinyl, 4-dihydropyranyl, and 3-morpholinyl; and groups such as tetrahydrofuran-2-ylmethyl and piperidin-3-ylethyl. “Hetetoalkanediyl” means 2 divalent group detived from a heteroalkyl by removal of a second hydrogen atom, such as 3-oxapentane-1,5-diyl Heteroalkyl and heteroalkanediyl groups also include those where a ring nitrogen is oxidized to form an

N-oxide. A “cycloamino” group is a cyclic heteroallcyl of 5 to 7 ring atoms containing a nitrogen ring atom by which the group is bonded to the remainder of the molecule of which it fotms a patt and optionally containing a further fing heteroatom selected from O, S, and NR (where Ris H ot C, , alkyl optionally substituted with halogen, hydroxy, ot 1 or 2 phenyl groups). 4-Methyl-1-piperazinyl, 4-(2-hydroxyethyl)-1-piperazinyl, 4-(diphenylmethyl)- 1-piperazinyl, and 4-morpholinyl are examples of cycloamino groups. Compounds of this invention also include compounds where any NR, group present is replaced by 2 cycloamino group. :

[0022] “Substituted heteroalkyl” and “substituted hereteroalkanediyl” mean heteroalkyl and heteroalkanediyl substituted in the manner described above for substituted alkyl

[0023] “Aryl” means 2 monovalent group derived from an aromatic hydrocarbon containing 6 to 14 ring carbon atoms by removal of one hydrogen atom from a carbon atom, which is monocyclic (e.g., phenyl), condensed polycyclic, for example, condensed bicyclic (e.g, naphthyl), or linked polycyclic, for example, linked bicyclic (e.g, biphenylyl). “Arenediyl” means 2 divalent group derived from an aryl by removal of a second hydrogen atom from a carbon atom, such as 1,4-benzenediyl, 1,5-naphthalenediyl, and biphenyl-4,4'-diyl. A preferred aryl is phenyl, and a preferred arenediyl is benzenediyl (any isomer).

[0024] “Substituted aryl” means aryl substituted with up to three substituents selected from halo, -CN, -NO,, —OR, optionally halo-substituted C, ; alkyl, optionally halo-substituted C, 5 alkyloxy, -SR, =COR, —OC(O)R, -C(O)OR, -NR,, -NR,'X, -PR,, -PR,;'X, -SO,OR,

-OSO,R, -SONR,, -NRSO.R, —CONR,, -NRCOR, and -NRC(O)OR, whete each Ris hydrogen, optionally R'-substituted alkyl, optionally R'-substituted heteroalkyl, optionally

R'-substituted aryl, optionally R'-substituted heteroaryl, optionally R'-substituted aralkyl, or optionally R'-substituted heteroaralkyl (preferably, hydrogen or optionally R'-substituted alkyl) and each R'is, independently, 1 to 3 substituents selected from halo, -CN, -NO,, -OH,

C, , alkyl, C, , alkyloxy, -SH, -NH,, or -C(O)Oalkyl (preferably, halo, -CN, -NO,, -OH,

C, , alkyl, C, ; alkyloxy, -SH, or -NH,), or two R groups form a 4- or 5-membex optionally

R'-substituted alkanediyl or optionally R'-substituted hetcroalkanediyl, and X is a halogen. Two adjacent substituents may also form a methylenedioxy or ethylenedioxy group. Substituted aryl groups include aryl groups substituted with up to three substituents selected from the group consisting of halo, -CN, -NO,, -OH, optionally halo-substituted C, , alkyl, optionally halo- substituted C, ; alkyloxy, -SH, and -NH,, for example, phenyl substituted in this way. “Substituted arenediyl’ means arenediyl substituted in the manner described above for an aryl. "Preferred substituted aryls are substituted phenyls.

[0025] “Aralkyl” means alkyl substituted with aryl, such as benzyl and phenethyl. A preferred aralkyl is benzyl. “Arenedialkyl” means two alkyls jointly substituted with atenediyl, such as benzene-1,4-dimethyl. A preferred arenedialkyl is benzenedimethyl (any isomer).

[0026] “Substituted aralkyl” means aralkyl in which one or both of the atyl and the alkyl are substituted in the manner described above for substituted aryl and substituted alkyl; and “substituted arenedialkyl” means arenedialkyl in which one or more of the arenediyl and the two alkyls are substituted in the manner described above for substituted arenediyl and substituted alkyl. The preferred substituted aralkyls are substituted benzyls; and preferred substituted arenedialkyls are substituted benzenedimethyls.

[0027] “Halogen” or “halo” means F, Cl, or Bx.

[0028] “Heteroaryl” means aryl in which 1 to 4 (preferably 1 to 3) of the ring carbon atoms are replaced by O, §, N, or NR (where R is H or C, alkyl), preferably O, S, or NR, including monocyclic groups containing 5 or 6 ring atoms such as furanyl, thienyl, pytrolyl, oxazolyl, imidazolyl, pyridinyl, pyrazinyl, pytidazinyl, pyrimidinyl, and the like, and bicyclic groups such as benzothiazolyl, purinyl, and benzimidazolyl. Monocyclic rings are preferred. “Heteroarenediyl” means a divalent group derived from a heteroaryl by removal of a second hydrogen atom from a carbon atom. Heteroaryl and heteroarenediyl groups also include those where 2 ring nitrogen is oxidized to form an N-oxide.

[0029] “Substituted heteroaryl” and “substituted heteroarenediyl” mean heteroaryl and heteroarenediyl substituted in the manner described above for substituted aryl.

[0030] “Heteroaralkyl” means alkyl substituted with heteroaryl, such as 2-thienylmethyl. “Heteroarenedialkyl” means two alkyls substituted jointly with heteroatenediyl, such as 2,5-furanyldiethyl.

[0031] “Substituted heteroaralkyl” and “substituted heteroatenedialkyl” mean heteroaralkyl and heteroarenedialkyl substituted in the manner described above for substituted alkyl and substituted arenedialkyl

[0032] “Salts” are described in the section entitled “Compounds of this invention”.

[0033] A “therapeutically effective amount” means that amount which, when administered to 2 human for treating a cancet, is sufficient to effect treatment for the cancer. “Treating” or “treatment” of a cancer in a human includes one or more of: (1 limiting/ inhibiting growth of the cancer, ie., limiting/arresting its development, 2 reducing/ preventing spread of the cancer, Le. reducing/preventing metastases, 3) relieving the cancer, ie., causing regression of the cancer, 4 reducing/pteventing recurrence of the cancer, and (5) palliating symptoms of the cancer.

[0034] “Combination therapy” means the administration of a corapound of the first or fourth aspects of this invention and another anticancer therapy during the course of cancer chemotherapy. Such combination therapy may involve the administration of the compound of the first ot fourth aspect of this invention before, during, and/or after the administration of the another anticancer therapy. The administration of the compound of the first or fourth aspect of this invention may be separated in time from the administration of the another anticancer therapy by up to several weeks, and may precede it or follow it, but more commonly the administration of the compound of the first or fourth aspect of this invention will accompany at least one aspect of the another anticancer therapy (such as the administration of one dose of a chemotherapeutic agent, molecular targeted therapy agent, biologic therapy agent, or radiation therapy) within up to 48 hours, and most commonly within less than 24 hours.

[0035] “Another anticancer therapy” is an anticancer therapy that is not a treatment with a compound of the first or fourth aspect of this invention. Such “another anticancer therapies” include chemotherapy; molecular targeted therapy; biologic therapy; and radiotherapy. These therapies ate those used as monotherapy or in combination therapy.

[0036] Chemotherapeutic agents include: alkylating agents, including: alkyl sulfonates such as busulfan, ethyleneimine derivatives such as thiotepa, nitrogen mustards such as chlorambucil, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, melphalan, and uramustine, . nitrosouteas such as carmustine, lomustine, and streptozocin, triazenes such as dacarbazine, procatbazine, and temozolamide, and platinum compounds such as cisplatin, carboplatin, oxaliplatin, satraplatin, and picoplatin; antimetabolites, including: antifolates such as methotrexate, permetrexed, raltitrexed, and ttimetrexate, purine analogs such as cladribine, chlorodeoxyadenosine, clofarabine, fludarabine, mercaptopurine, pentostatin, and thioguanine, pyrimidine analogs such as azacitidine, capecitabine, cytarabine, edatrexate, floxuridine, fluorouracil, gemcitabine, and troxacitabine; natural products, including: antitumor antibiotics such as bleomycin, dactinomycin, mithramycin, mitomycin, mitoxantrone, porfiromycin, and anthracyclines such as daunorubicin (including liposomal daunorubicin), doxotubicin (including liposomal doxorubicin), epirubicin, idarubicin, and valrubicin, enzymes such as L-aspataginase and PEG-L-asparaginase, microtubule polymer stabilizers such as the taxanes paclitaxel and docetaxel, mitotic inhibitors such as the vinca alkaloids vinblastine, vinctistine, vindesine, and vinorelbine, topisomerase I inhibitors such as the camptothecins irinotecan and topotecan, and topoisomerase II inhibitors such as amsactine, etoposide, and teniposide; hormones and hormone antagonists, inchiding: androgens such as fluoxymesterone and testolactone,

antiandrogens such as bicalutamide, cyproterone, flutamide, and nilutamide, aromatase inhibitors such as aminoglutethimide, anastrozole, exemestane, formestane, and letrozole, corticosteroids such as dexamethasone and prednisone, estrogens such as diethylstilbestrol, antiestrogens such as fulvestrant, raloxifene, tamoxifen, and toremifine,

LHRH agonists and antagonists such as buserelin, goserelin, leuprolide, and triptorelin, progestins such as medroxyprogesterone acetate and megestrol acetate, and thyroid hormones such as levothyrozine and liothyronine; and miscellaneous agents, including altretamine, arsenic trioxide, gallium nitrate, hydroxyurea, levamisole, mitotane, octreotide, procarbazine, suramin, thalidomide, lenalidomide, photodynamic compounds such as methoxsalen and sodium pozfimer; and proteasome inhibitors such as bortezomib.

[0037] Molecular targeted thetapy agents include: funcdonal therapeutic agents, including: gene therapy agents, antisense therapy agents, : tyrosine kinase inhibitors such as etlotinib hydrochloride, gefitinib, imatinib mesylate, and semaxanib, and gene expression modulators such as the retinoids and rexinoids, e.g. adapalene, bexaroteae, trans-retinoic acid, 9-s-retinoic acid, and N-(4-hydroxyphenyl)retinamide; phenotype-directed therapy agents, including: monoclonal antibodies such as alemtuzumab, bevacizumab, cetuximab, ibritumomab tiuxetan, rituximab, and trastuzumab, immunotoxins such as gemtuzumab ozogamicin, radioimmunoconjugates such as *'I-tositumomab, and cancer vaccines.

[0038] Biologic therapy agents include: interferons such as interferon-a,, and interferon-a,, and interleukins such as aldesleukin, denileukin diftitox, and oprelvekin.

[0039] In addition to these agents intended to act against cancer cells, anticancer therapies include the use of protective or adjunctive agents, including:

cytoprotective agents such as amifostine, dexrazoxane, and mesna, phosphonates such as pamidronate and zoledronic acid, and stimulating factors such as epoetin, darbeopetin, filgrastim, PEG-filgrastim, and sargramostim.

[0040] Combination cancer therapy regimens with which the compounds of the fitst aspect of this invention may be combined include all regimens involving the use of two or more of the anticancer therapies (anticancer agents) such as those mentioned in paragraphs [0036] to

[0038] above and/or radiotherapy, optionally including protective and adjunctive agents such as those mentioned in patagraph [0039] above; and the compound of the first or second aspect of this invention can be added to existing anticancer regimens known for the treatment of various cancers, such as the regimens mentioned in such books as Cancer Chemotherapy and

Biotherapy: Principles and Practice, 3rd ed. (2001), Chabner and Longo, eds., and Handbook of

Cancer Chemotherapy, 6th ed. (2003), Skeel, ed., both from Lippincott Williams & Wilkins,

Philadelphia, Pennsylvania, U.S.A; and regimens for anticancer therapies, especially chemotherapies, may be found on Web sites such as those maintained by the National Cancer

Institute (www.cancet.gov), the American Society for Clinical Oncology (www.asco.org), and the National Comprehensive Cancer Network (www.nccn.otg).

[0041] Many combination chemotherapeutic regimens are known to the art, such as combinations of platinum compounds and taxanes, e.g. catboplatin/ paclitaxel, capedcitabine/ docetaxel, the “Cooper regimen”, fluorouracil-levamisole, fluorouracil- leucovorin, methotrexate-leucovorin, and those known by the acronyms ABDIC, ABVD, AC,

ADIC, AL, BACOD, BACOP, BVCPP, CABO, CAD, CAE, CAF, CAP, CD, CEC, CF,

CHOP, CHOP+rituzimab, CIC, CMF, CMFP, CyADIC, CyVADIC, DAC, DVD, FAC,

FAC-S, FAM-S, FOLFOX-4, FOLFOX-6, M-BACOD, MACOB-B, MAID, MOPP, MVAC,

PCV, T-5, VAC, VAD, VAPA, VAP-Cyclo, VAP-II, VBM, VBMCP, VIP, VP, and the like.

[0042] Combinations of chemotherapies and molecular targeted therapies, biologic therapies, and radiation therapies are also well known to the art; including therapies such as trastuzumab + paclitaxel, alone or in further combination with carboplatin, for certain breast cancers, and many other such regimens for other cancers; and the “Dublin regimen” and “Michigan regimen”, both for esophageal cancer, and many other such regimens for other cancets.

[0043] “Comprising” or “containing” and their grammatical variants are words of inclusion and not of limitation and mean to specify the presence of stated components, groups, Steps, and the like but not to exclude the presence or addition of other components, groups, Steps, and the like. Thus “comprising” does not mean “consisting of”, “consisting substantially of”, or “consisting only of”; and, for example, a formulation “comprising” a compound must contain that compound but may also may contain other active ingredients and/or excipients.

[0044] Compounds of this invention

[0045] In a first aspect, this invention is sulfonylethyl phosphorodiamidates, compounds of formula A, B, and C:

Sd ed

R 0 R ATR

R, 9 R - R oo R : db R _S0, -

A 0,5 R? J rR" 80; a R O R py Mn

RK § R RoR

CA © B C where: each R is independently hydrogen, C, alkyl, or -CH,CH,X, where each X is independently Cl

Bt, C, , alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, Cy, alkyl, halo-C,, alkyl, C,, alkyloxy, or halo-C, ; alkyloxy, provided that at least two R’s in each phosphorodiamidate group are —-CH,CH,X;

R! is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; and R*is optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally substituted arenediyl, optionally substituted arenedialkyl, optionally substituted heteroarenediyl, or optionally substituted heteroarenedialkyl, and their salts.

[0046) In a fourth aspect, this invention is thioethyl phosphotodiamidates, compounds of formula BB and CC:

R J R g iy R re” (

J ‘

R§ R y— hand <8 k

BB cC where: each R is independently hydrogen, C, ¢ alkyl, or ~CH,CH,X, where each X is independently Cl,

Bt, C, alkanesulfonyloxy, halo-C, ¢ alkanesulfonyloxy, ot benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, ; alkyl, halo-C, ; alkyl,

C, alkyloxy, ot halo-C, , alkyloxy, provided that at least two R’s in each phosphorodiamidate group are —-CH,CH,X;

R'is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; and

R? is optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally substituted arenediyl, optionally substituted arenedialkyl, optionally substituted heteroarenediyl, or optionally substituted heteroarenedialkyl, and their salts.

[0047] Salts (for example, pharmaceutically acceptable salts) of the compounds of formulae

A, B, C, BB, and CC ate included in the present invention and are useful in the compositions, methods, and uses described in this application (see Berge et al., J. Phar. Sea., 66:1 (1971) fora nonexclusive list of pharmaceutically acceptable salts).

[0048] These salts include salts that may be formed when acidic protons present are capable of teacting with inorganic or organic bases. Typically the parent compound is treated with an excess of an alkaline reagent, such as hydroxide, carbonate or alkoxide, containing an appropriate cation, Cations such as Na*, K*, Ca®*, Mg? and NH," are exatnples of cations present in pharmaceutically acceptable salts. Suitable inorganic bases, therefore, include calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide. Salts may also be prepared using organic bases, such as salts of primary, secondary and tertiary amines, substituted amines including paturally-occurting substituted amines, and cyclic amines including isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, tromethamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, N-alkylglucamines, theobtomine, purines, piperazine, piperidine, N-ethylpiperidine, and the like.

[0049] If 2 compound of the first or fourth aspects of this invention contains a basic group, such as an amino or phosphino group, it may be prepared as an acid addition salt. Acid addition salts of the compounds are prepared in a standard manner in a suitable solvent from the patent compound and an excess of an acid, such as hydrochlotic acid, hydrobromic acid, sulfuric acid (giving the sulfate and bisulfate salts), nitric acid, phosphoric acid and the like, and organic acids such as acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, salicylic acid, 4-toluenesulfonic acid, hexanoic acid, heptanoic acid, cyclopentanepropionic acid, lactic acid, 2-(4-hydroxybenzoyl)benzoic acid, 1,2-cthanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2.Joct-2-ene-1-carboxylic acid, glucoheptonic acid, gluconic acid, 4,4'-methylenebis(3-hydroxy-2-naphthoicjacid, 3-phenylpropionic acid, trimethylacetic acid, #r7-butylacetic acid, laurylsulfuric acid, ghucuronic acid, glutamic acid, 3-hydroxy-2-naphthoic acid, steatic acid, muconic acid and the like.

[0050] Certain of the compounds may form inner salts or zwitterions.

[0051] Certain compounds of formula A, B, and BB of the invention tnay contain one or more chiral centers (for example, those based on amino acids). In such cases, all individual stereoisomers and racemic and non-tacemic mixtures of the stereoisomers of the compounds (and their salts) are included in this invention. Typically the stereochemistry of the compounds of this invention will be dictated by the stereochemistry of the thiols of formula A’ aad B', and methods for the isolation of individual stereoisomers of such thiols, for example, the resolution of thiol-amino acids such as cysteine, ate well known to a person of ordinary skill in the art and ate therefore not described here. In,many instances, such as with cysteine and penicillamine, individual stereoisomers and racemic mixtures of stereoisomers are commercially available; and other thiols such as dithiothreitol and dithioerythritol exist as individual stereoisomets.

[0052] The compounds are named in this application semi-systematically based on the CAS name of canfosfamide (i.e. with the thioethy! or sulfonylethyl phosphorodiamidate portion of the molecule named as a substituent on the temainder of the molecule). Thus, the compounds 70A, 128A, 23B, and 1C below:

Cl Cl

EnV Vand ci cl o oo i a Ma hn: iad pa Wa N of 9 ps

Yo hat oF

SO, S02 L, o_ p) A wp A ci N ci J © RNa ~~ cl cl 70A 128A ~~ 8 cl cl 1c 238 ate named 3-[[2-[[bis[bis(2-chloroethyl)amino]phosphinyljoxy]ethyljsulfonyl]-L-alanine, a-[[2-[[bis[bis(2-chloroethyljamino]phosphinyljoxy] ethyl}sulfonyl]toluene, 1,5-di[[2-[[bis[bis(2-chloroethyl)amino]phosphinyljoxy]ethyljsulfonyllnaphthalene, and di[2-[[bis[bis(2-chloroethyl)amino]phosphinyl]oxyjethyl sulfone, respectively.

[0053] Compounds of this invention include those compounds of formulae A, B, C, BB, and

CC where one or more of the following is true: 1. each R is -CH,CH,X; 2. each X is Cl, Bt, methanesulfonyloxy, ttifluoromethanesulfonyloxy, benzenesulfonyloxy, or 4-toluenesulfonyloxy; especially where X is Cl ot Bt; particularly Cl; 3. the compound is a compound of formula A; 4. R! is substituted with one or mote groups that enhance the solubility of the compound over a compound that is not so substituted, for example, hydroxy, carboxy, sulfo, amino, quaternary ammonium, phosphino, and quaternary phosphonium;

S. R' is optionally substituted alkyl, or optionally substituted aryl, or optionally substituted aralkyl; and 6. R'is the residue of an optionally protected thiol-amino acid, or an elaborated thiol- amino acid such as an amide or ester formed by teaction at the carboxy group and/or amide ot sulfonamide formed by reaction at the amine group, preferably the residue of a thiol-amino acid; 7. R! is the residue of 2-mercaptoacetic acid or 3-mercaptopropionic acid, or an elaborated acid such as an amide or ester formed by reaction at the catboxy group, particularly the residue of an optionally elaborated 2-mercaptoacetamide; 8. the compound is a compound of formula B; 9. R? is substituted with one ot mote groups that enhance the solubility of the compound over a compound that is not so substituted, for example, hydroxy, carboxy, sulfo, amino, quaternary ammonium, phosphine, and quaternary phosphonium; or 10. the compound is 2 compound of formula C.

[0054] Compounds of this invention include each of the compounds described in the specification and claims of this application as filed, including in the Examples below, especially compounds 5A, 13A, 154, 23A, 704, 128A, 171A, 172A, and 180A, and their salts.

Compositions, uses, and methods of this invention include compositions, uses, and methods where the compound is one of those mentioned in the preceding sentence.

[0055] Preparation of the compounds

[0056] Sulfonylethyl phosphorodiamidates of formula A, B, and C may conveniently be prepared by oxidation of the corresponding thioethyl phosphorodiamidates of formulae AA,

BB, and CC (with any reactive moiety in R' ot R? protected against oxidation if necessary), as follows:

Ruy R Roy R CL Roy R aS . RoR

R S~o-b=0 obo" R S~~"o-p=0 o=p-0"" ~"o-p=0 rR RNR rR RR rR

AA BB cC

Emme solvent 0, RoR RoR 0, 0 Re-R Ray R o, RoR

RIS o-p=0 o=p-0">" TRS o-p=0 0=p-0"" S"o-p=0

RR rR rR RNR RR

A B ©

[0057] This oxidation may be performed by any of the methods known in the att for the oxidation of thioethers to sulfones, such as the use of peracids (peroxycatboxylic acids), persulfates, perborates, peroxides, ozone, iodosyl reagents, halogens, and the like. If the compound of formula AA or BB contains a reactive moiety in R' or R?, such as an amine group, that reactive moiety may be protected before the oxidation and the resulting protected compound of formula A or B deprotected if desired to yield the final compound (though the protected compounds are also among compounds of this invention). Such protection and deprotection is seen in Synthetic Example 2, for example. Where a peracid is used, a typical procedure involves dissolving the compound of formula AA, BB, or CC in a solvent such as acetic acid or isopropyl acetate at reduced temperature, followed by the addition of the peracid (e.g. peracetic acid) in excess.

[0058] After oxidation and optional deprotection, the compounds of formula A and formula

B may be elaborated by synthetic methods known per sz, as discussed later in this application.

[0059] The thioethyl phosphorodiamidates of formula AA and BB may conveniently be prepated by one of two methods: (1) reaction of the corresponding thiols of formula A! and B' with a 2-X'-ethyl . phosphorodiamidate (where X' is Cl, Br, or an alkane- or atenesulfonyloxy group such as methanesulfonyloxy, benzenesulfonyloxy, or 4-toluenesulfonyloxy), as follows:

R1-SH HS—R?—SH

Al B’

Roy ® y o=p—0""" rR

RR R. .R R., .R

RS oibmo MEN S—RE=S._~g_ho

N N N

RR R™ TR R™ TR

AA BB ; and (2) (a) reaction of the thiols of formula A! and B’ with a 2-X'-ethanol (where X' is Cl, Br,

C, ; alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, , alkyl, halo-C, ; alkyl, C,,alkyloxy, or halo-C,, alkyloxy) to first form the corresponding 2-hydroxyethyl thiols of formula A? and B?, followed by (b) reaction of the 2-hydroxyethyl thiols with a phosphorodiamidyl halide or alkane- or arenesulfonate (3 is Cl, Br, C, ; alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, , alkyl, halo-C, ; alkyl, C, ; alkyloxy, or halo-C, ; alkyloxy), as follows:

R1-SH HS—R2?-SH

Al B! x1

HO” —R2-

R= _~ou Ho” ~~ R Soon

A2 B?

Ro nr - N ~

R R

<..R « -R RL, .R

R'-S " | A \ S—R?-8 \ ~"o-p=0 o=p-0" ~"0-p=0 rr rr Nr

AA BB }

[0060] The direct conversion of thiols of formula A! and B' to the thioethyl phosphorodiamidates of formula AA and BB may be performed by any of the methods known in the att for alkylation of thiols. A typical procedure involves dissolving the thiol of formula A! or B! in a polar solvent such as water, an alcohol, dimethylformamide, ot tetrahydrofuran, then treating it with a base such as a hydroxide, alkoxide, fluoride, or tertiary amine or amide base to form the thiolate anion, followed by adding the phosphorodiamidate.

Thiolate displacement of the leaving group X! of the 2-X'-ethyl phosphorodiamidate gives the compounds of formula AA and BB.

[0061] Alternatively, the thiols of formula A! and B' may be treated with ethanol 2_substituted with CL, Br, C,, alkanesulfonyloxy, halo-C, 4 alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo,

C,; alkyl, halo-C, ; alkyl, C, ; alkyloxy, or halo-C, ; alkyloxy, e.g. 2-chloroethanol, under the conditions of the previous paragraph to give the 2-hydroxyethylthiol compounds of formula

A? and B? which may then be converted to the cornpounds of formula AA and BB, typically by reacting the compounds of formula AZ? and B? with a phosphorotliamidyl halide or alkane- or arenesulfonate in an aprotic solvent such as tetrahydrofuran, toluene, or dichloromethane in the presence of an base such as an alkoxide or tertiary amine.

[0062] After optional deprotection, the compounds of formula AA and formula BB may be elaborated by synthetic methods known per s¢, as discussed later in this application.

[0063] The compounds of formula CC are conveniently prepared by a two-step synthesis, in which 2,2'-thiodiethanol is reacted with a sub-stoichiometric amount of a phosphotodiamidyl halide or alkane- or arenesulfonate in an aprotic solvent in the presence of a base such as an alkoxide to give the mono-phosphorodiamidate ester, and that ester is then reacted with an excess of the phosphorodiamidyl halide or alkane- or arenesulfonate, again in an aprotic solvent in the presence of a base such as an alkoxide, to give the bis-phosphorodiamidate ester, as follows:

R O R

Rw® rR OR Ray R N—P—N

OH X2-P=0 NPN x2-P=0 R 0 R § Mor en Mor §

S — § — S

Pp 5

HO RAI cc

[0064] Pharmaceutical compositions and administration

[0065] The second and fifth aspects of this invention are pharmaceutical compositions comprising 2 compound of the first or fourth aspect of this invention and optionally a pharmaceutically acceptable excipient.

[0066] The compounds of the first and fourth aspects of this invention may be administered by any route suitable to the subject being treated and the nature of the subject's condition.

Routes of administration include administration by injection, including intravenous, intraperitoneal, intramuscular, and subcutaneous injection, by transmucosal or transdermal delivery, through topical applications, nasal spray, suppository and the like or may be administered orally. Formulations may optionally be liposomal formulations, emulsions, formulations designed to administer the drug across mucosal membranes or transdermal formulations. Suitable formulations for each of these methods of administration may be found, for example, in Remington: The Science and Practice of Pharmacy, 20th ed., A. Gennaro, ed,

Lippincott Williams & Wilkins, Philadelphia, Pennsylvania, U.S.A. Typical formulations will be cither oral or solutions for intravenous infusion. Typical dosage forms will be tablets or capsules for oral administration, solutions fot intravenous infusion, and lyophilized powders for reconstitution as solutions for intravenous infusion.

[0067] Depending on the intended mode of administration, the pharmaceutical compositions may be in the form of solid, semi-solid ot liquid dosage forms, preferably in unit dosage form suitable for single administration of a precise dosage. In addition to an effective amount of the active ingredients), the compositions may contain suitable pharmaceutically-acceptable excipients, including adjuvants which facilitate processing of the active compounds into preparations which can be used pharmaceutically. “Pharmaceutically acceptable excipient” refers to an excipient or mixture of excipients which does not interfere with the effectiveness of the biological activity of the active ingredient(s) and which is not toxic to the host to which it is administered.

[0068] For solid compositions, conventional excipients include, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, magnesium carbonate, and the like. Liquid pharmacologically administrable compositions can, for example, be prepared by dissolving, dispetsing, etc., an active compound as described herein and optional pharmaceutical adjuvants in an excipient, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, to theteby form a solution or suspension. If desited, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary excipients such as wetting or emulsifying agents, pH buffering agents and the like, for example, sodium acetate, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, etc.

[0069] For oral administration, the composition will generally take the form of a tablet or capsule, ot it may be an aqueous or nonaqueous solution, suspension ot syrup. Tablets and capsules are preferred oral administration forms. Tablets and capsules for oral use will generally include one or more commonly used excipients such as lactose and corn starch.

Lubricating agents, such as magnesium stearate, are also typically added. When liquid suspensions are used, the active agent may be combined with emulsifying and suspending excipients. If desired, flavoring, coloring and/or sweetening agents may be added as well.

Other optional excipients for incorporation into an otal formulation include preservatives, suspending agents, thickening agents, and the like. —23_

[0070] Injectable formulations can be prepared in conventional forms, either as liquid solutions ot suspensions, solid forms suitable for solubilization ot suspension in liquid ptiot to injection, or as emulsions or liposomal formulations. The sterile injectable formulation may also be a sterile injectable solution or a suspension in a nontoxic patenterally acceptable diluent or solvent. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils, fatty esters or polyols are conventionally employed as solvents or suspending media.

[0071] The pharmaceutical compositions of this invention may also be formulated as lyophilized powders for parenteral administration. Powders may be reconstituted by addition of water or other aqueous medium and then further diluted with a suitable diluent prior to use.

The liquid formulation is generally a buffered, isotonic, aqueous solution. Examples of suitable diluents are isotonic saline solution, 5% dextrose in water, and buffered sodium or ammonium acetate solution. Pharmaceutically acceptable solid or liquid excipients may be added to enhance or stabilize the composition, ot to facilitate preparation of the composition.

[0072] Typically, 2 pharmaceutical composition of the present invention is packaged ina container with a label, ot instructions, ot both, indicating use of the pharmaceutical composition in the treatment of cancer.

[0073] The pharmaceutical composition may additionally contain one or more other pharmacologically active agents in addition to a compound of this invention. These additional active agents will typically be useful in treating cancet, of for enhancing the treatment of cancer by compounds of this invention.

[0074] Uses of the compounds

[0075] The compounds of the first and fourth aspect of this invention have activity against human cancer cell lines, as demonstrated in the #7 vw#o and in vivo Examples below, and are 95 therefore considered to be useful as human cancer chemotherapeutic agents, for the treatment of human cancers.

[0076] Thus, the third and sixth aspects of this invention include methods of treating cancer in humans by administering a therapeutically effective amount of a compound of the first or fourth aspect of this invention, or a pharmaceutical composition of the second or fifth aspect of this invention, to the human; and the use of compounds of the first and fourth aspects of this invention in the manufacture of medicaments for the treatment of cancer in humans.

Optionally, the method may further comprise treating the human with another anticancer therapy, such as a therapy already conventional for the cancer being treated.

[0077] Cancers that ate particulatly treatable by the method of this invention are cancers with sensitivity to inducers of apoptosis, and more specifically those cancers that express of, particularly, overexpress one or mote glutathione S-transferase isoenzymes. Cancers that express Of OVercgpress one or more glutathione S-transferase isoenzymes when treated with other anticancer compounds or combination cancet chemotherapy regimens ate especially treatable by the method of this invention. Such cancers include cancers of the brain, breast, bladder, cervix, colon and rectum, esophagus, head and neck, kidney, lung, liver, ovary, pancreas, prostate, and stomach; leukemias such as ALL, AML, AMML, CLL, CML, CMML, and hairy cell leukemia; Hodgkin’s and non-Hodgkin's lymphomas; mesotheliomas, multiple myeloma; and sarcomas of bone and soft tissue. Cancers particularly treatable by the method of this invention include breast, ovarian, colorectal, and non-small cell lung cancets.

[0078] The amount of the compound of the first or fourth aspect of this invention that is administered to the human (either alone of, more usually, in a composition of the second or fifth aspect of this invention) should be a therapeutically effective amount when used alone or when used in conjunction with the another anticancer therapy (if the compound of the first or fourth aspect of this invention is administered in conjunction with another anticancer therapy); and similarly the amount of the another anticancer therapy that is administered to the mammal (if the compound of the first or fourth aspect of this invention is administered in conjunction with another anticancer therapy) should be a therapeutically effective amount when used in conjunction with the compound of the first or fourth aspect of this invention.

However, the therapeutically effective amount of either the compound of the first or fourth aspect of this invention and the amount of the another anticancer therapy when administered fe combination cancer chemotherapy may each be less than the amount which would be therapeutically effective if delivered to the human alone, It is common in cancer therapy, * though, to use the maximum tolerated dose of the or each therapy, with a reduction only because of common toxicity of the therapies used or potentiation of the toxicity of one therapy by another. Because of the lack of cross-resistance of canfosfamide, for example, with several common chemotherapeutic agents, and its relative lack of clinically severe toxicity, especially its lack of clinically severe hematological toxicity, it is expected that compounds of the first and fourth aspect of this invention will be administrable at essentially their maximum tolerated dose as a single agent, and no reduction in the amount of the another anticancer therapy will be required.

[0079] The compounds of the first and fourth aspects of this invention, or pharmaceutical compositions of the second and fifth aspects of this invention, are thus used to treat cancer in humans requiring such treatment, by administering 2 therapeutically effective amount of the chosen compound or composition. Therapeutically effective amounts of compounds of the invention are in the range of 10 - 10,000 mg/m?, for example, 30 - 3000 mg/m’ ot 100 - 1000 mg/m? Dosing may be at 1 - 35 day intervals; for example, about 500 - 1000 mg/m? at 1 - 5 week intervals, especially at 1, 2, 3, or 4 week intervals, or at highest frequencies including as frequently as once/day for several (e.g. 5 or 7) days, with the dosing repeated every 2, 3, ot 4 weeks, ot constant infusion for a petiod of 6 — 72 hours, also with the dosing repeated every 2, 3, or 4 weeks. Suitable dosages and dose frequencies will be readily determinable by a person of ordinary skill in the art having regard to that skill and this disclosure. No unacceptable toxicological effects are expected when compounds of the invention ate administered in accordance with the present invention.

[0080] Suitable dosing for the other anticancer therapy (if the compound of the first ox fourth aspect of this invention is used in combination) will be the dosing already established for that therapy, as described in such documents as those listed in paragraph [0040]. Such dosing varies widely with the therapy: for example, capecitabine (2500 mg/m” orally) is dosed twice daily for 2 weeks on and 1 week off, imatinib mesylate (400 or 600 mg/day orally) is dosed daily, rituximab is dosed weekly, paclitaxel (135 - 175 mg/m?) and docetaxel (60 - 100 mg/ m’) atc dosed weekly to every three weeks, catboplatin (4 - 6 mg/ ml_emin) is dosed once every 3 or 4 weeks (though the doses may be split and administered over several days), nitrosourea alkylating agents such as carmustine are dosed as infrequently as once every 6 weeks.

Radiotherapy may be administered as frequently as weekly (or even within that split into smaller dosages administered daily).

[0081] A person of ordinary skill in the art of cancer thetapy will be able to ascertain a therapeutically effective amount of the compound of the first oz second aspect of this invention and a therapeutically effective amount of another anticancer therapy for a given cancer and stage of disease without undue experimentation and in reliance upon personal knowledge and the disclosure of this application.

[0082] Combination therapies include the combination administration of a compound of the first aspect of this invention with a platisum compound such as catboplatia ot cisplatin, optionally in further combination with gemcitabine or a taxane such as docetaxel or paclitaxel; with gemcitabine; with a taxane; with an anthracycline such as doxorubicin or liposomal doxorubicin; with oxaliplatin, optionally in further combination with capecitabine or fluorouracil /leucovorin; and with gemcitabine or a platinum compound such as carboplatin or cisplatin, in further combination with a vinca alkaloid such as vinorelbine.

[0083] Examples

[0084] The following examples illustrate the preparation of compounds of this invention, and their activity in predictive i #ifro and in vivo anticancer assays.

[0085] Synthetic Examples.

[0086] The compounds of this invention are prepared by conventional methods of organic chemistry. See, for example, Larock, “Comprehensive Organic Transformations”, Wiley-VCH,

New Yotk, New York, U.S.A. In some cases, protective groups may be introduced and later removed. Suitable protective groups for amino, hydroxyl, and carboxyl groups are described in

Greene et al. “Protective Groups in Organic Synthesis”, 2nd ed., 1991, John Wiley and Sons,

New York, New York, U.S.A. The compounds of this invention can be synthesized, generally following the synthetic schemes illustrated earlier in this application, as shown in the following examples or by modifying the exemplified syntheses by means known to those of ordinaty skill in the art.

[0087] The synthetic examples below show compounds where R in formulae A, B, C, BB, and CC are all 2-chloroethyl. It will be apparent that compounds where R is other than 2-chlotoethyl may be produced by patallel methods: for example: (a) compounds where R is 2-bromoethyl may be made by reacting bis(2-btomoethyl)amine,

POC], and ethylene glycol to produce 2-hydroxyethyl tetrakis (2-bromoethyl)- phosphorodiamidate, esterifying to an appropriate sulfonate ester, and using that to alkylate the thiols of formula A® and B! or 2,2'-thiodiethanol to form the compounds of formulae AA,

BB, and CC; (b) compounds where R is 2-(Z-sulfonyloxy)ethyl may be made by using bis(2-hydroxyethylamine) instead of bis(2-bromoethylamine) in the process of (2) above, then fotming the sulfonate esters; and

Claims (32)

We claim:

1. A compound of formula A, formula B, or formula C R © R Np R, R R 0 R N RN, pbx ’ R R [0] 3 RZ” SO; S 0, - 0,8 J Na C R 9 R ay A 0 NPN NN R R d ) Rr Oo A B Cc whete: each Ris independently hydrogen, Cy alkyl, ot -CH,CH,X, where each X is independently Cl, Br, C,, alkanesulfonyloxy, halo-Cy alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, alkyl, halo-C, 5 alkyl,

C,., alkyloxy, or halo-C,; alkyloxy, provided that at least two R’s in each phosphorodiamidate group ate —-CH,CHX; R'is optionally substituted alkyl, optionally substituted heteroalkyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optionally substituted heteroaralkyl; and R? is optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally substituted arenediyl, optionally substituted arenedialkyl, optionally substituted heteroarenediyl, ot optionally substituted heteroarenedialkyl, or its salt.

2. The compound of claim 1 where each R is -CH,CHX.

3. The compound of claim 2 where each X is Cl, Br, methanesulfonyloxy, trifluoromethanesulfonyloxy, benzenesulfonyloxy, or 4-toluenesulfonyloxy.

4. The compound of claim 3 where each Xis Cl

5. The compound of any one of claims 1 to 4 that is a compound of formula A or its salt.

6. The compound of claim 5 where R! is substituted with one or mote groups that enhance the solubility of the compound over a compound that is not so substituted.

7. The compound of claim 5 where R' is optionally substituted alkyl.

8. The compound of claim 5 where R' is the residue of an optionally protected thiol- amino acid, or an elaborated thiol-amino acid, preferably the residue of a thiol-amino acid.

9. The compound of claim 5 where R'is an optionally elaborated 2-metcaptoacetic acid ot 3-metcaptopropionic acid, preferably the residue of an optionally elaborated 2-mercaptoacetamide.

10. The compound of claim 5 or 6 whete R! is optionally substituted aralkyl

1. The compound of claim 10 where R! is optionally substituted benzyl.

12. The compound of claim 1 that is selected from: (2-[bis(bis(2-chlorocthyDaminojphosphinyloxylethyljsulfonylbutane, N-methyl-2-[[2-[[bis{bis (2-chloroethylamino]phosphinyljoxylethylisulfonyllacetamide, N-{2-[[2-[[bis|bis(2-chloroethyl) amino]phosphinyljoxy] cthyl}sulfonyljacetyl} L-phenylalanine N-[2-(4-morpholino)ethyl]-2-{[2- [[bis[bis(2-chloroethyl) amino]phosphinyljoxylethyl] sulfonyl]- acetamide, 3-[[2-[{bis [bis(2-chloroethyl)amino]phosphinyljoxyjethyl) sulfonyl]-L-alanine, o-{[2-[{bis[bis (2-chloroethyl)amino] phosphinyljoxyjethyl] sulfonyljtoluene, a-[[2-[[bisbis(2-chloroethylamino] phosphinyljoxyjethyljsulfonyl}-4- {[4-(2-thenoyl)- 1-piperazinyljcarbonyl} toluene, -[[2-[[bis[bis(2-chloroethyDamino]phosphinyljoxylethyllsulfonyl-4-[(ethoxycacbonylmethy)- aminocarbonyl]toluene, and 2-[[[2-{bis[bis(2-chloroethyljaminolphosphinyl] oxyletbyl]sulfonyljmethyljthiophene, and their salts.

13. The compound of claim 12 that is N-methyl-2-[[2-[[bis[bis (2-chloroethyl)amino]- phosphinylioxylethyl] sulfonyllacetamide.

14. The compound of claim 12 that is 3.{[2-[[bis[bis(2-chloroethylamino]phosphinyljoxy]- ethyl]sulfonyl]-L-alanine or its salt.

As, I (EEA PCT/US2005/017880

15. The compound of any one of claims 1 to 4 that is a compound of formula B or its salt.

16. The compound of claim 15 where R’ is substituted with one or more groups that enhance the solubility of the compound over a compound that is not so substituted.

17 The compound of any one of claims 1 to 4 that is a compound of formula C or its salt.

18. A pharmaceutical composidon comprising a compound of any one of claims 1 to 17.

19. The use of a compound of any one of claims 1 to 17 in the manufacture of a medicament for the treatment of cancer.

20. A method of preparing a compound of formula A, formula B, or formula C: R OO R NPN R § R R 0 R NN R o R R oo R wh - r R R SO oO R? 2 SO, - 0,8 J SO ~ R17 iS R O R " 1 % N—P—N Th R R = u - 0 A B Cc where: each R is independently hydrogen, C, alkyl, or -CH,CH,X, where each X is independently Cl, Br, C, ; alkanesulfonyloxy, halo-C, ( alkancsulfonyloxy, or benzencsulfonyloxy optionally substituted with up to three substituents selected from halo, C, ; alkyl, halo-C, ; alkyl, C,;alkyloxy, or halo-C, ; alkyloxy, provided that at least two R’s in cach phosphorodiamidate group are —~CH.CH.X R'is opdonally substituted alkyl, optionally substituted heteroalkyl, optionally substtuted aryl, optionally substituted aralkyl, optionally substituted heteroaryl, or optonally substituted heteroaralkyl; and -74 - AMENDED SHEET

PCT/US2005/017880 R? is optionally substituted alkanediyl, optionally substituted heteroalkancdiyl, optionally substituted arenediyl, optionally substituted arenedialkyl, optonally substituted heteroarenediyl, or optionally substituted heteroarencdialkyl, orits salt, comprising: (a) oxidaton of the corresponding compound of formula AA, formula BB, or formula CC: Oo eben “i { R oo R / RN R 0 R / R oo R N=P—N / R Oo R _S I R17 S R O R pA) pone Ro R RoR AA BB CC with any reactive moiety in R' or R? protected against oxidation if necessary; optionally followed by one or more of: (b) deprotection of a protected compound of formula A, formula B, or formula C; (©) conversion of the R groups of a compound of formula A, formula B, or formula C into other R groups to form another compound of formula A, formula B, or formula C; (d) elaboration of the R' group of 2 compound of formula A or the R* group of the compound of formula B by synthetic methods known per se to form another compound of formula A or formula B; (e) formation of a salt of a compound of formula A, formula B, or formula C; 63) conversion of a salt of a compound of formula A, formula B, or formula C to another salt of formula A, formula B, or formula C; and (g conversion of a salt of a compound of formula A, formula B, or formula C to a non-salt form of the compound of formula A, formula B, or formula C. _75- CLEAN COPY

PCT/US2005/017880

21. A compound of formula BB or formula CC: R OO R NPN AY GC R R TUN J RK 6 R J g _R? S j g r R O R iY 7 g N=P=N, Th R R R 0 R © BB CC where: each Ris independenty hydrogen, C, alkyl, or —~CH,CH.X, where each X is independently Cl, Br, C, ( alkanesulfonyloxy, halo-C, ; alkanesulfonyloxy, or benzenesulfonyloxy optionally substituted with up to three substituents selected from halo, C, ; alkyl, halo-C, 5 alkyl, C, ; alkyloxy, or halo-C, , alkyloxy, provided that at least two R’s in each phosphorodiamidate group arc ~CIH.CH.Xj and R%is optionally substituted alkanediyl, optionally substituted heteroalkanediyl, optionally subsatuted arenediyl, optionally substituted arenedialkyl, optionally subsututed heteroarenediyl, or opuonally substituted heteroarenedialkyl, or its salt.

22. The compound of claim 21 where cach R is ~-CH,CH,X.

23. The compound of claim 22 where cach X 1s Cl, Br, methanesulfonyloxy, trifluoromethanesulfonyloxy, benzenesulfonyloxy, or 4-toluencsulfonyloxy.

24. The compound of claim 23 where each X is Cl.

25. The compound of any one of claims 21 to 24 that is a compound of formula BB or its salt. = 76 - AMENDED SHEET

PCT/US2005/017880

26. ‘The compound of any one of claims 21 to 24 that is a compound of formula CC or its salt.

27. A pharmaceutical composition comprising a compound of any one of claims 21 to 26.

28. The use of a compound of any one of claims 21 to 26 in the manufacture of a medicament for the treatment of cancer.

29. A compound according to any one of claims 1 to 17 or 21 to 26, substantially as herein described with reference to and as illustrated in any of the examples.

30. A composidon according to claim 18 or claim 27, substantially as hercin described with reference to and as illustrated in any of the examples.

31. Use according to claim 19 or claim 28, substantally as herein described with reference to and as illustrated in any of the examples.

32. A method according to claim 20, substanually as herein described with reference to and as illustrated in any of the examples. -77- AMENDED SHEET