NZ571028A - Nitrobenzindole compounds and their use in cancer treatment - Google Patents

Nitrobenzindole compounds and their use in cancer treatment

Info

Publication number
NZ571028A
NZ571028A NZ571028A NZ57102808A NZ571028A NZ 571028 A NZ571028 A NZ 571028A NZ 571028 A NZ571028 A NZ 571028A NZ 57102808 A NZ57102808 A NZ 57102808A NZ 571028 A NZ571028 A NZ 571028A
Authority
NZ
New Zealand
Prior art keywords
formula
indole
compound
mmol
benzo
Prior art date
Application number
NZ571028A
Inventor
Moana Tercel
Ralph James Stevenson
Shangjin Yang
Ho Huat Lee
Jean-Jacques Youte Tendoung
Frederik Bastiaan Pruijn
Original Assignee
Auckland Uniservices Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Auckland Uniservices Ltd filed Critical Auckland Uniservices Ltd
Priority to NZ571028A priority Critical patent/NZ571028A/en
Priority to PCT/NZ2009/000184 priority patent/WO2010027280A1/en
Publication of NZ571028A publication Critical patent/NZ571028A/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/56—Ring systems containing three or more rings
    • C07D209/58—[b]- or [c]-condensed
    • C07D209/60—Naphtho [b] pyrroles; Hydrogenated naphtho [b] pyrroles
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/553—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom having one nitrogen atom as the only ring hetero atom
    • C07F9/572—Five-membered rings
    • C07F9/5728—Five-membered rings condensed with carbocyclic rings or carbocyclic ring systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
    • C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
    • C07F9/02—Phosphorus compounds
    • C07F9/547—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom
    • C07F9/6558—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system
    • C07F9/65583—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing at least two different or differently substituted hetero rings neither condensed among themselves nor condensed with a common carbocyclic ring or ring system each of the hetero rings containing nitrogen as ring hetero atom

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Disclosed are nitrobenzindole compounds of formula (I) or pharmaceutically salts thereof, wherein Z is a sidechain of formula (Ia), (Ib), (Ic) or (Id) and the other substituents are disclosed within the specification. The use of the said compounds for the preparation of a medicament for treating cancer is also disclosed.

Description

New Zealand Paient Spedficaiion for Paient Number 571 028 Received at IPONZ on 14 December 2010 NITROBENZINDOLES AND THEIR USE IN CANCER THERAPY TECHNICAL FIELD The present invention relates generally to nitro-l,2-dihydro-3.H-benzo[<?]indoles and related analogues, to their preparation, and to their use in cancer therapy.
BACKGROUND TO THE INVENTION It has been established that many human tumors contain a significant proportion of hypoxic cells (Kennedy et al., Int.], Radiat. Oncol. Biol. Pbjs., 1997, 37, 897-905; Brown et al., Cancer Res. 1998, 58, 1408-1416; Vaupel et al., Semin. Oncol., 2001, 28, 25-35). The presence of hypoxic cells arises because of chaotic growth and an inefficient microvasculature system within the tumour, so that tumours often exhibit large inter capillary distances and variable blood flow. Reduction of oxygen tension in 15 tumors leads to radioresistance. Up to a three-fold increase in radiation dose may be required to kill anoxic tumor cells. A link has been identified between the presence of tumor hypoxia and failure of local control by radiation therapy (Nordsmark et al., Radiother. Oncol, 1996, 41, 31-39; Brizel et al., RMdiolher. Oncol., 1999, 53, 113-117; Movsas et al., Uroksj/, 2002, 60, 634-639; Koukourakis et al.,/. Clin. Oncol, 2006, 24, 727-735). Hypoxia also contributes to chemoresistance by mechanisms which 20 include limitation of the delivery of drugs to hypoxic regions of tumors (Minchinton & Tannock, Nat. Ren Cancer, 2006, 6, 583-592; Hicks et al., /. Natl Cancer Inst., 2006, 98, 1118-1128). The phenomenon of tumor hypoxia has been exploited tn the development of a class of anticancer agents termed 'hypoxia-activated prodrugs' which are also sometimes referred to as 'bioreductive drugs' although the latter term also encompasses prodrugs activated by reduction under oxic 25 conditions (Brown et al, Semin. Radiat. Oncol., 1966, 6, 22-36; Denny ct al., Br. J. Cancer, 1996, 74 (Suppl. XXVII) 32-38; Stratford & Workman, Anti-Cancer Drug Des., 1998, 13, 519-528; Brown et al., Nat. Rep. Cancer 2004, 4, 437; Brown et al., frontiers in Bioscience, 2007, 12, 3483).
Various mtro(hetero)aromatic compounds have been reported as hypoxia-activated prodrugs. These 30 include • the nitroimidazole (i), which is proposed to undergo fragmentation following nitro group reduction by endogenous cellular nitroreductase enzymes (McClelland et al., Biochem. Pharmacol,, 1984, 33, 303-309), Received at IPONZ on 14 December 2010 • the dinitrobenzamidc mustard (ii) and analogues, where similar reduction of the nitro group activates the mustard (Palmer et al., J. Med. Cbem. 1996, 39, 2518; Helsbv et al., Chem. Rfx. Toxicol., 2003, 16, 469-478; Wilson et al., Radiation Res. 2007,167, 625; Patterson et al, Clin. Cancer Res. 2007, 13, 3922; Denny et al., NZ Patent 529249), and • the nitrobeiizo[c]indole (ili) and analogues have been reported as potential bioreductive drugs activated by the B. coli NTR enzyme (Denny et al., PCT Int. Appl. WO 98/11101 A2, 1998; Atwell et alOrg. Cbem. 1998, 63, 9414-9420; Atwell et al., Bioorg. Med. Chem. Tett, 1997, 7, 1493-1496.) • Certain nitrobenzo[tf]indoles and analogues thereof, and their use in cancer therapy, are also disclosed in PCT international application PCT/NZ2005/000278 (WO 2006/043839).
It is an object of die present invention to provide a specific class of nitro-l,2-dihydro-3H-benzoMindoles, that are useful in cancer treatment, or to at least Drovide the oublic with a useful it' ' i. i alternative.
SUMMARY OF THE INVENTION In a first aspect, the present invention provides a compound of Formula I, Y or N02 wherein Y is selected from CI or Br, and wherein X is selected from S02NR'2 or CONR1,, where X is located at either position 7 and where each R1 independently represents PI or a Q 4alkyl, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb J Received at IPONZ on 14 December 2010 (la) (lb) (ic) (id) wherein R2 represents a lower C,_4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) 5 substituent, and R' represents H, C1.4alkyl, CMalkoxy, CM alkynyl or CMalkynyloxy and pharmaceutically acceptable salts thereof.
In certain embodiments, R3 represents H or C,_4alkyl.
In certain embodiments, R'1 represents H.
In certain embodiments, R'1 represents C,_4 alkynyl or C, 4 alkynyloxy, where the alkyne functional group is a terminal alkyne.
In certain embodiments, R3 is selected from the following groups: In certain embodiments, Y is CI.
In certain embodiments, Y is Br.
In certain embodiments, X is located at the 7 position.
In certain embodiments, X is7-SO,NH(CH2)2OH.
In certain embodiments, X is 7~CONH(CH2)2OH.
In certain embodiments, Z is selected from the group consisting of: Received at IPONZ on 14 December 2010 In certain embodiments, R1 is H, Y is CI or Br, and X is 7-S02NH(CFI2)20H or 7-SO,NH(CH,).,OH. In such embodiments, Z may be selected from groups A to D above. Alternatively, Z may be a group la, wherein R2 is C2 or C3 alkyl bearing a morpholine (Ic) 5 substituent.
In certain embodiments, the compound of Formula I is selected from the group consisting of: l-(Chloromethyl)-A'-(2-bydroxyethyl)-3-[5-(2-moi-pholinoethoxy)-lH-indole-2-carbonyl]-5-mtro-l,2-dihydro-3H-benzo[>]indole-7-sulfonamide; 1 - (Chloromethyl) -A- (2-hydroxye thyl) -3 - [5- (3-morphoIinopropoxy)-1 H-indole-2-carbonyl] -5-nitro-l,2-dihydro-3.H-benzo[?]indole-7-sulfonamide; (£)-l-(Chloi-omethyl)-A'-(2-hydroxyethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acrylovl}-5-nitro-l,2-dihydro-3H-benzo(V]indole-7-sulfonamide; l-(Chloromethyl)-A'-(2-hydroxyethyl)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-l ,2-15 dihydro-3/-/-benzo[i?Jindole-7-sulfonamide; l-(Chloromethyl)-A?-(2-hydroxyethyl)-3-[5-(2'-morpholinoethoxy)-l.H-indole-2-carbonyl]-5-nitro-l ,2-dihydro-3 H-benzo [ej indolc-7-carboxamide; (E)-l - (chloromethyl)-A-(2-hydroxyethyl)-3- {3- [4-(2-morpholinoethoxy)phenyl]acryloyl} -5-nitro-l ,2-dihydro-3jFir-benzo[fi']indole-7-carboxamidc; 20 (i)-l-(Cliloromethyl)-A-(2-hydroxyethyl)-3-[5-(2-morpholinoethox}r)-l H-indole-2-carbonyl]-5-nitro-l,2-dihydro-3H-benzo[e]indole-7-sulfonamide; l-(Bromomethyl)-A'-(2-hydroxyethvl)-3-[5-(2-morpholinoethoxy)-lH-indole-2-carbonyl]-5-nitro-l,2-dihydro-3 H-benzo [i?]indole-7-sulfonamide; and (.K)-l-(Bromomethyl)-A-(2-hydroxyethyl)-3- {3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-25 1,2-dihydro-3.H-benzo[<?]indole-7-sulfonamide.
In a second aspect, the present invention provides a compound of Formula II, Received at IPONZ on 14 December 2010 (ii) nh2 wherein Y is selected from CI or Br, and wherein X is selected from SO,NR'2 or CONR'2, where X is located at either position 7 or 8, and where each R1 independentiy represents H or a Cl.4alkyl, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb (la) (lb) (Ic) (Id) wherein R2 represents a lower C2_4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) substituent, and R1 represents H, a C14 alkyl, C14alkoxy, CM alkynyl or CU4 alkynyloxy, and phannaceutically acceptable salts thereof.
In certain embodiments, R3 represents H or C, ,, alkyl.
In certain embodiments, R"1 represents H.
In certain embodiments, R' represents C,.4 alkynyl or C,.4 alkynyloxy, where the alkyne functional group is a terminal alkyne.
In certain embodiments, R'1 is selected from the following groups: In certain embodiments, Y is CI.
In certain embodiments, Y is Br.
In certain embodiments, X is located at the 7-position.
Received at IPONZ on 14 December 2010 In certain embodiments, X is 7-SO,NH(CH,)2OH.
In other embodiments, X is 7-CONH(CH2)2OH.
In certain embodiments, Z is selected from the group consisting of: ^ n ° // o CK .
B N O C In certain embodiments, R' is H, Y is CI or Br, and X is 7-S02NH(CH2)20H or 7- S02NH(CH,)20H. In such embodiments, Z may be selected from groups A to C above. Alternatively, Z may be a group la, wherein R2 is C2 or C3 alkyl bearing a morpholine (Ic) substituent.
"I T fi -l-nm l"\ /A 11-h-i it #-o +-1-1 r~ i>-i mirf /~\ £ T-<" ✓"H-H-v-i > 11 ii TT 1 c o ■£■*•✓"■»+vs -J-V*/> rtivn n t-\ s-f\ «-i c s c #-1 +-"i ,-v <".-£• XwJ AAA ^UlMVUUJl^lJWj Ultu ^VlllpUUllVi KJ L _L VIX« A A J.O OV1VV WJ., -Amino-1 -(chloromethyl) - N- (2-hydroxyethyI)-3-[5-(2-morpholinoethoxy) -1 H-indole-2-carbony Ij -ls2-dihydro-3.H-benzo[<?]indole-7-sulfonamide; -Amino-1 -(chloromethyl)-A;-(2-hydroxyethyl)-3-[5-(3-morpholinopropoxy)-1 H-indole-2-carbonyl] -l,2-dihvdro-3 H-benzo (VJindole-7-siilfonamide; 20 (E)-5-Amino-l-(chloromethyi)-N-(2-hydroxyethyl)-3- {3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-l,2-dihydro-3H-benzo[e]indole-7-sulfonamide; -Amino-l-(chloromethyl)-A/-(2-hydroxyethyl)-3-[5-(2-morpholinoethoxy)-l H-indole-2-carbonylJ-l,2-dihydro-3i 7-benzo[<?]indole-7-caiboxamide; and (JT)-5-Amino-l-(chloromethyl)-iV-(2-hydroxyethyI)-3-[5-(2-morpholinoethoxy)-l J /-indole-2-25 carbonyl]-1,2-dihvdro-3H-benzo [Vjindole-7-sulfonamide.
In a third aspect, the present invention provides compounds of Formula Illa-d, Received at IPONZ on 14 December 2010 Formula 1 0 1 1 Formula 1 —0 r r4 i i or —OP(OH)2 or N-- Formula II Formula II .0 HJ (Ilia) (lllb) Formula I or Formula IE D , R -O ( Formula I or Formula II TRq 9 'OP(OH)2 (IHc) (Mid) wherein Formula I and Formula II and the subsituents thereof are as defined above, and any compound of Formula I or Formula II bearing a free hydroxyl group is functionalised with a phosphate (Formula Ilia) or an amino acid or short polypeptide chain (Formula lllb) or a monosaccharide (Formula IIIc), or wherein any compound of Formula I or Formula II bearing a free hydroxyl group or a secondary or tertiary amine is functionalised with a phosphonooxymethvl substituent (Formula Hid), wherein R4 is selected from any substituent found in the naturally occuring amino acids, m is selected from 1, 2, 3, or 4, Rs represents any combination of hydroxy, hydroxymethyl, carboxylic acid or other substituents found in naturally occuring monosaccharides, p represents 0 or 1, T represents O or N, and when T represents N it represents the secondary amine of the indole la or the tertiary amine of the morpholine Ic or either of the tertiary amines of the piperazine Id, wherein R represents the remainder of the structure of Formula I or Formula II compatible with these definitions, and q represents 0 when T represents O and q represents 1 or 2 when T represents N, and when q represents 2 the charge of the quaternary ammonium group of Formula Illd is balanced by an appropriate anion, and pharmaceutical!)- acceptable salts thereof.
In certain embodiments, the compound of Formula III is a compound of Formula Ilia.
In certain embodiments, R3 represents H or C|_4 alkyl.
In certain embodiments, R3 represents H.
In certain embodiments, R'' represents CM alkynyl or CU4 alkynyloxy, where the alkyne functional 25 gi'oup is a terminal alkyne.
Received at IPONZ on 14 December 2010 In certain embodiments, R' is selected from the following groups: ^ \ In certain embodiments, Y is CI.
In ccrtain embodiments, Y is Br.
In certain embodiments, X is 7-SO,NH(CH2),OH. In such embodiments X niav be functionalized with a phosphate group to form the group 7-S02NH(CH2)20P0{0H)2 .
In other embodiments, X is 7-CONH(CH2)2OH. In these embodiments, X may be functionalized with a phosphate group to form the group 7-CONH(CH2)2OPO(OH)2 In certain embodiments, Z is selected from the group consisting of: In certain embodiments, the compound of Formula Ilia is selected from the group consisting of: 2-[l-(Chloromethvl)-3-[5-(2-morpholinoethoxy)-lJH-indolc-2-carbonyl]-5-nitro-l,2-diliydro-3.H-20 benzo[£']indole-7-sulfonamido]ethyl dihydrogen phosphate; 2- [1 -(Chloromethyl)-3- [5-(3-morpholinopropoxy)-l H-indole-2-carbonyl] -5-nitro-l ^-dihydro-SH-benzo[i?]indole-7-sulfonamido]ethyl dihydrogen phosphate; (Ej-2-(l -(chloromethyl)-3- {3- |4-(2-morpholinoethoxy)phenyl] acryloyl} -5-nitro-l ,2-dihvdro-3.H-benzo[e]indole-7-sulfonamido)ethy.l dihydrogen phosphate; 25 2-[l-(Chloromethyl)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-l,2-dihydro-3.H-benzo[i?]indole-7-sulfonamido]ethyl dihydrogen phosphate; 2- {1 -(Chloromethyl)-3~[5-(2-niorpholinoethoxy)-l H-indole-2-carbonyl]-5-nitro-1,2-dihydro-3H-benzo[V|mdole-7-carboxamido} ethyl dihydrogen phosphate; Received at IPONZ on 14 December 2010 (£)-2-[l -(Chloromcthyl)-3- {3-[4-(2-morpholinoethoxy)phenyl]acrvloyl} -5-nitro-l ,2-dihvdro-3 H-benzo[t']indole-7-carboxamido]ethyl dihydrogen phosphate; (j)-2-[l-(Chloromethyl)-3-[5-(2-morphoIinoethoxy)-l.H-indole-2-carbonyl]-5-nitro-ls2-dihydro-3H-benzo[tf]indole-7-sulfonamido] ethyl dihydrogen phosphate; (S,E)-2-[1 -(Chloromethyl)-3- {3- [4-(2-morpholinoethoxy)phenyl]acryloyl} -S-nitro-l ,2-dihydro-3.H-benzo[i?]iiidole-7-sulfonamido]ethyl dihydrogen phosphate; 2-[l-(Bromometh)'l)-3-[5-(2-morpholinoethoxy)-l.H-indole-2-carbonyl]-5-nitro-l,2-dihydro-3i-/-benzo[#]indole-7-sulfonamido]ethyl dihydrogen phosphate; and (E)-2-[l -(Bromomethyl)-3- {3- [4-(2-morpholinoethoxy)phenylJacryloyl} -5-nitro-l ,2-dihydro-3H-10 benzo[f]indole-7-sulfonamido]ethyl dihydrogen phosphate.
In a further aspect, the present invention provides a pharmaceutical composition comprising a compound of Formula I, II or III as defined above or a pharmaceutical!}' acceptable salt thereof and a pharmaceutical!)' acceptable carrier.
Also described herein is a method for the production of an anti-cancer effect in a warm-blooded animal such as a human, wherein the method comprises administering to the animal an effective amount of a compound of Formula I, II or III as defined above or a pharmaceutical!)' acceptable salt thereof.
Also described herein is a method for the production of an anti-cancer effect in a cell, wherein the method comprises contacting the cell with an effective amount of a compound of Formula I, II or III as defined above or a pharmaceutically acceptable salt thereof. Anti-cancer effects include, but are not limited to, anti-tumour effects, the response rate, the time to disease progression and the 25 survival rate. Anti-tumour effects include but are not limited to, inhibition of tumour growth, tumour growth delay, regression of tumour, shrinkage of tumour, increased time to regrowth of tumour on cessation of treatment and slowing of disease progression.
Also described herein is a method for the treatment of a cancer in a warm-blooded animal such as a 30 human, which comprises administering to said animal an effective amount of a compound of Formula I, II or III as defined above or a pharmaceutically acceptable salt thereof.
In a further aspect, the present invention provides the use of a compound of Formula I, II or III as defined above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for 35 the production of an anti-cancer effect in a warm-blooded animal such as a human.
Received at IPONZ on 14 December 2010 In a further aspect, the present invention provides the use of a compound of Formula I, II or III as defined above or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a cancer in a warm-blooded animal such as a human.
In further aspects, the invention provides methods of preparing compounds of the general Formulae I, II or III, as defined above. Such methods are described below.
"Effective amount" means an amount of a compound that, when administered to a subject for 10 treating a disease state, is sufficient to effect such treatment for the disease state. The "effective amount" will vary depending on the compound, disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.
"Treating" or "treatment" of a disease state includes: (i) preventing the disease state, i.e. causing the clinical symptoms of the disease state not to develop in a subject that may be exposed to or predisposed to the disease state, but does not yet experience or display symptoms of the disease state, (ii) inhibiting the disease state, or (iii) relieving the disease state, i.e. causing temporary or permanent regression of the disease state or its clinical symptoms.
"Pharmaceutically acceptable" means that which is useful in preparing a pharmaceutical composition 25 that is generally safe, non-toxic, and neither biologically nor otherwise undesirable and includes that which is acceptable for veterinary as well as human pharmaceutical use.
"Pharmaceutically acceptable salts" of a compound means salts that are pharmaceutically acceptable, as defined herein, and that possess the desired pharmacological activity of the parent compound. 30 Such salts include: acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like; or formed with organic acids such as acetic acid, methanesulfonic acid, maleic acid, tartaric acid, citric acid and the like; or salts formed when an acidic proton present in the parent compound either is replaced by a 35 metal ion, e.g. an alkali metal ion, an alkaline earth ion, or an aluminium ion; or coordinates with an Received at IPONZ on 14 December 2010 organic or inorganic base. Acceptable organic bases include ethanolamine, dicthanolamine, N-methylglucamine, triethanolamine and the like. Acceptable inorganic bases include aluminium hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate and sodium hydroxide.
"Warm blooded animal" means any member of the mammalia class including, but not limited to humans, non-human primates such as chimpanzees and other apes and monkey species, farm animals such as cattle, horses, sheep, goats, and swine; domestic animals such as rabbits, dogs and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like.
Compounds of Formulae I, II or III as defined above may exist in different enantiomeric and/or diastereomeric forms. In such cases it is to be understood that Formulae I, II and III include any possible enantiomeric or diastereomeric forms, and mixtures of such forms, and also any pharmaceutically acceptable salts thereof.
In certain embodiments, the compound of Formula I, II or III may be in the form of a racemic mixture. In other embodiments, the compound of formula I, II or III may be in the form of a mixture in which either the or the R enantiomer predominates, or in the form of either the j" or the R enantiomer substantially free of the other enantiomer, for example containing less than about 5%, such as less than about 1%, of the other enantiomer.
While the invention is broadly as defined above, further aspects of the invention will become apparent with reference to the following description, reaction schemes and examples.
BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 shows an ORTEP representation of the crystal structure of (R)-di-/«r/-butyl 2-[l-(chloromethyl)-5-nitro-3-(trifluoroacety])-l ,2-dihydro-3H-benzo[(?]indole-7-sulfonamido]ethyI phosphate (144).
Figure 2 shows an ORTEP representation of the crystal structure of (J)-di-/f?7-butyl 2-[l- (chloromethyl)-5-nitEo-3-(tiifluoroacetj4)-l,2-dihydro-3/-/-benzo[(?]indole-7-sulfonamido]ethyl phosphate (145). 12 Received at IPONZ on 14 December 2010 Figure 3a is a bar graph comparing the activity in SiHa xenografts in combination with radiation of certain compounds of this invention and reference compounds, some of which are described in WO 2006/043839.
Figure 3b is a bar graph comparing the activity in H460 xenografts in combination with radiation of certain compounds of this invention and reference compounds, some of which are described in WO 2006/043839.
DETAILED DESCRIPTION OF THE INVENTION Compounds of the invention As defined above, this invention relates to nitrobenziiidoles and related analogues, in particular aminobenzindoles, of the Formulae I, II or III as defined above, and their use in treating cancer.
The invention more particularly relates to the use of these compounds as prodrugs that are activated under hypoxic conditions, ie in an environment having a lower oxygen tension than that of normal tissues, to treat cancer.
The compounds of the present invention have the same core ring structure as the compounds disclosed in WO 2006/043839. However, the compounds of the present invention differ from those disclosed in WO 2006/043839 in that they incorporate an indole or cinnamate group Z that bears an O-linked sidechain containing a morpholine or N-methylpiperazine substituent.
Examples of compounds of the invention of the Formulae I, II and III, respectively, are set out in Tables 1 to 3 below.
Received at IPONZ on 14 December 2010 Table 1: Examples of Compounds of Formula I of the Invention Y z = no2 -W, O. -ay-— h b rfY°^0 fY°^0 c d No X y z Enantiomeric form 1 7-SO,NH(CH2),OH CI A 2 7-SO,NH(CH2),OH CI B RS 3 7-SO,NH(CH2)2OH CI C K5" 4 7-S02NH(CH2)20H CI D RS 7-CONH(CH2)2OH CI A RS 6 7-CONH(CH2),OH CI C RS 7 7-S02NH(CH2)20H CI A S 8 7-S02NH(CH2)20H Br A RS 9 7-SO,NH(CH2)2OH Br C RS Tabie 2: Jixampies of Compounds of formula II of the Invention Y z = ° nh2 h b c No X y z Enantiomeric form 7-S02NH(CH2),0H CI A RS 11 7-SO,NH(CH2)2OH CI B RS 12 7-S02NH(CH2),0H CI C RS 13 7-CONH(CH2)2OH CI A RS 14 7-S02NH(CH2),0H CI A S Received at IPONZ on 14 December 2010 Table 3: Examples of Compounds of Formula III of the Invention Y kit a A. /N^Z .vyy t z= x7Vy 0 NO 2 H B [TY°^0 ifY^O C D No X Y Z Enantiomeric form 7-S02NH(CHa)20P0(0H)2 CI A Kf 16 7-SO,NH(CH2)2OPO(OH)2 CI B RJ 17 7-S02NH(CH2)20P0(0H)2 CI C RS" 18 7-S02NH(CH2),0P0(0H)2 CI D RS 19 7-CONH(CH2)2OPO(OH)2 CI A RS 7-CONH(CH2)2OPO(OH)2 CI C RS 21 7-S02NH(CH2)20P0(0H)2 CI A S 22 7-S02NH(CH2)20P0(0H)2 CI C S 23 7-S02NH(CH2)20P0(0H)2 Br A RS 24 7-S02NH(CH2)20P0(0H)2 Br C RS Received at IPONZ on 14 December 2010 Methods of preparation of compounds of the invention The compounds of Formulae I, II and III of the present invention may be prepared using the methods described below. These methods form further aspects of the invention. The schemes A to 5 M following the general descriptions of the methods below include details of the reagents used to achieve each of the synthetic steps.
Preparation of compounds of Formula I Compounds of Formula I may be prepared by first preparing an intermediate compound of Formula IV, V or VI as defined below. These compounds can be converted to desired compounds of Formula I by the methods following the description of the preparation of these intermediate compounds.
Compounds of Formula IV Br x- (IV) wherein X is defined for a compound of Formula I and J represents /mf-butoxycarbonyl or trifluoroacetyl, can be prepared by a method including in a first step reacting a compound of Formula V CI X NJ (V) wherein X is defined for a compound of Formula I and J represents /<?r/-butoxycarbonyl or mfluoroacetyl, with an effective amount of an iodide salt in a non-nucleophilic solvent, to provide a compound of Formula VI Received at IPONZ on 14 December 2010 wherein X is defined for a compound of Formula I and J represents AvZ-butoxycarbonyl or tiifluoroacetyl, and in a sccond step reacting a compound of Formula VI with an effective amount of a silver sulfonate salt to provide a compound of Formula VII wherein X is defined for a compound of Formula I, J represents /mf-butoxycarbonyl or tiifluoroacetyl, and Rr' represents C, 4 alkyl or benzyl or benzyl substituted with C,_4 alkyl or nitro groups, and in a third step reacting a compound of Formula VII with an effective amount of a bromide salt in a non-nucleophilic solvent, to provide a compound of Formula IV.
Preferably the first step is achieved using sodium iodide in methyl ethyl ketone.
Preferably the sccond step is achieved using silver mesylate in acetonitrile.
Preferably the third step is achieved using lithium bromide in tetrahydrofuran.
Compounds of Formula IV as defined above may also be prepared by the reaction of compounds of Formula V with an effective amount of a bromide salt in a non-nucleophilic solvent.
Compounds of Formula V as defined above can be prepared by the methods described in WO 2006/043339, the full content of which is incorporated herein by reference.
Compounds of Formula I may be prepared from the above compounds of Formulae IV and V by a method which includes in a first step reacting a compound of Formula VIII Y wherein X and Y are as defined for a compound of Formula I and J represents PI, tert-butoxycarbonyl or tiifluoroacetyl, with an effective amount of a nitrating agent to provide a compound of Formula IX 0S02R6 nj X [T (VII) Received at IPONZ on 14 December 2010 Y X NJ (IX) N02 wherein X and Y are as defined for a compound of Formula I and J represents FI or tiifluoroacetyl, and in a second step for the cases where ] of Formula IX represents tiifluoroacetyl reacting a compound of Formula IX with an effective amount of a weak base in the presence of water to 5 provide a compound of Formula IX where J represents FI, and in a third step reacting a compound of Formula IX where J represents FI with a compound of Formula ZCO,FI or ZCOC1 wherein Z is as defined for a compound of Formula I, to provide a compound of Formula I.
Preferably the first step is achieved using potassium nitrate in concentrated sulfuric acid. 10 Preferably the second step is achieved using cesium carbonate in aqueous methanol.
Preferably when a compound of Formula ZCO,H is used in the third step then this step is achieved using A-(3-dimethylaminopropyl)-Ap-ethyl carbodiimide or l-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methiodide or other suitable coupling reagent.
Preferably when A'-(3-dimethylaminopropyl)-A!'-ethyl carbodiimide or l-[3-(dimethylamino)propyl]-3-ethylcarbodiimide methiodidc is used in the third step then an effective amount of an acid co-reagent such as toluenesulfonic acid is also used.
Preferably when a compound of Formula ZCOC1 is used in the third step then this compound is formed by the reaction of a compound of Formula ZCO,H with thionyl chloride or oxalyl chloride or oxalyl chloridc in the presence of catalytic dimethylformamide.
Preferably when a compound of Formula ZCOC1 is used in the third step then this step is achieved 25 using at least one equivalent of non-nucleophilic base such as di-zio-propylethylamine.
Compounds of the Formula ZC02FE can be prepared by a method which includes in a first step reacting a compound of Formula ZCO,ll' wherein R' represents C1-4 alkyl or phenyl or benzyl optionally substituted with one or more halide substkuents, and Z is as defined for a compound of 30 Formula I and contains the group Q wherein Q represents OH or NFI2 or C02PI, with a suitable derivative of a compound of Formula R2 where R2 is as defined for a compound of Formula I, and Received at IPONZ on 14 December 2010 includes die structures Ie-In, such suitable derivatives being alcohols or halides or primary or secondary amines or acids or acid chlorides. Compounds of formula ZC02R can be readilv prepared using methods known to those skilled in the art.
When Q represents OH or NH, the reaction may involve alkylation with a halide derivative of R2, when Q represents OH or an aromatic ring the reaction may involve a Mitsunobu reaction of an alcohol derivative of R2, when Q represents NH, the reaction may involve acylation using an acid chloride derivative of R2 or amide coupling using an acid derivative of R2, when Q represents CO,FI the reaction may represent amide coupling with a primary or secondary amine derivative of R2.
Additionally, when Q represents a halogen, suitable examples of ZCO,R may be formed by palladium or other metal catalysed reactions with halide derivatives of R2. The second step of the method includes reacting a compound of Formula ZCO,R' with base to provide a compound of Formula ZC02H.
Preferably the second step is achieved using potassium hydroxide or lithium hydroxide or sodium hydroxide in an aqueous alcoholic solvent.
Compounds of Formula I wherein X represents S02NR'2 and R1 is as defined for a compound of 20 Formula I, may be prepared by reacting a compound of Formula I wherein X represents S02C1 with a primary or secondary amine derivative of R1.
Preferably the reaction is conducted in the presence of at least one equivalent of a non-nucleophilic base such as di-ww-propylethylamine or triethylamine.
Compounds of Formula I wherein X represents CONR1, and R1 is as defined for a compound of Formula I, may be prepared by reacting a compound of Formula I wherein X represents CO,FI with a primary or secondary amine derivative of R1, using an effective amount of a suitable coupling reagent.
Preferably the reaction is conducted using (benzotriazoFl-yloxy)tripyrrolidinophosphonium hexafluorophosphate.
Preparation of compounds of Formula II jg Received at IPONZ on 14 December 2010 Compounds of the Formula II may be prepared by reducing a compound of Formula I under suitable conditions.
Preferably the reduction is achieved by hvdrogcnation using a suitable metal catalyst such as 5 platinum or palladium, or bv reaction with a metal such as zinc, preferably in a finely divided or powdered state, in the presence of a weak acid such as ammonium chloride.
Preparation of compounds of Formula III Compounds of Formula Ilia may be prepared by a method that includes in a first step reacting a compound of Formula I or Formula II bearing a free hydroxyl group with an effective amount of a phosphorylating agent to provide a compound of Formula Hie O —op(or8)2 (I lie) wherein RB represents CM alkyl or benzyl or substituted benzyl, and the phosphorylation reaction 15 may be a one-step process using an effective amount of an appropriate phosphoramidite or a two™ ' step process using phosphorus oxychloride followed by reaction of the intermediate with an alcohol R8OPI, and in a second step reacting a compound of Formula Ille with an appropriate reagent to provide a compound of Formula Ilia, which reagent may be an acid when R8 represents tert-hxitx\, or hydrogen and a metal catalyst when RK represents benzyl or substituted benzyl.
Preferably when Rs represents tert-butyl the first step is achieved using di-iW-butyl N,N-diethylphosphoramidite or di-/w/-butyl A/jA-di-zVo-propylphosphoramidite.
Preferably when Rs represents /<?r/-butyl the second step is achieved using trifluoroacetic acid.
Compounds of Formula Ilia may also be prepared by reacting alcohol derivatives of R1 or Formula IX using the phosphorylating conditions as described, and incorporating the so-formed phosphate ester into the general reactions as described above, to provide an alternative route to compounds of Formula Ille.
Formula I or Formula II Compounds of formula lllb may be prepared by a method including in a first step reacting a compound of Formula I or Formula II bearing a free hydroxyl group with an amino terminal- Received at IPONZ on 14 December 2010 protected amino acid or dipeptide or tripeptide or tetrapeptide using an effective amount of a peptide coupling reagent to provide a compound of Formula Illf Formula 1 r r4 i i or —o yn- Formula II [o HJ m (Illf) wherein R4 and m are as defined for a compound of Formula lllb and R9 represents a suitable 5 amino protecting group such as /i?r/-butoxycarbonyl or 9-fluorenylmethoxycarbonyl, and in a second step reacting a compound of formula Illf with an appropriate reagent to provide a compound of Formula lllb, which reagent may be an acid when R9 represents /?r/-butoxycarbonyl or a base when RJ represents 9-fluorenylmethoxycarbonyl.
Preferably the first step is achieved using 0-(benzotriazol-l-yl)-A',A?,A'',A1'-tetramethyluronium hexafluorophosphate or O-^-azabenzotriazol-l-y^-ATjAfJVVV-tetramethylui-onium hexafluorophosphate.
Preferably when R9 represents /W-butoxycarbonyl the second step is achieved using trifluoroacetic 15 acid.
Preferably when R'; represents 9-fluorenylmethoxycarbonyl the second step is achieved using morpholine or piperidine.
For those examples of compounds of Formula Mb where R4 contains a reactive functional group such as an amino group or carboxylic acid group, then suitably protected forms of these groups such as are commonly used in peptide synthesis may be employed, which protecting groups may be removed in the second step as described or in a prior or subsequent deprotection step to provide a compound of Formula lllb.
Compounds of Formula lllb may also be prepared by reacting alcohol derivatives of R1 or Formula IX using the peptide coupling conditions as described, and incorporating the so-formed amino terminal-protected amino acid or dipeptide or tripeptide or tetrapeptide into the general reactions as described above, to provide an alternative route to compounds of Formula Illf.
Compounds of Formula IIIc may be prepared by a method that includes in a first step reacting a compound of Formula I or Formula II bearing a free hydroxyl gioup with a suitably protected Received at IPONZ on 14 December 2010 21 monosaccharide donor such as a monosaccharide bromide or monosaccharide imidate to provide a compound of Formula Hlg of mono- or polysaccharides, and in a second step or steps reacting a compound of Formula Illg with an appropriate reagent to provide a compound of Formula IIIc, which reagent may be an ester-protected carboxylic acid.
Preferably when the first step is achieved using a monosaccharide bromide an effective amount of a silver salt such as silver triflate or silver carbonate is also used.
Preferably when the first step is achieved using a monosaccharide imidate then a tnchloroacetimidate is used, and further an effective amount of a Lewis acid such as boron Preferably when R1" represents an acetate-protected hydroxy or hydroxymethyl group the second 20 step is achieved using lithium hydroxide or potassium hydroxide in an aqueous alcoholic solvent.
Compounds of Formula IIIc may also be prepared by reacting alcohol derivatives of R1 or Formula IX with the monosaccharide donors as described, and incorporating the so-formed suitably protected monosaccharides into the general reactions as described above, to provide an alternative 25 route to compounds of Formula IHg.
Compounds of Formula Illd may be prepared by a method that includes in a first step reacting a compound of Formula I or Formula II bearing a free hydroxyl group or a secondary or tertiary amine group with an effective amount of a compound of Formula X Formula I or Formula II p aqueous base when R1" represents an acetate-protected hydroxy or hydroxymethyl group or a methyl R11^0P(0R12)2 (X) Received at IPONZ on 14 December 2010 22 wherein Rn represents a good leaving group which may be a halide or a sulfonate such as p-toluenesulfonate or methanesulfonate or trifluoromethanesulfonate and R12 represents a suitable phosphate protecting group which may be alkyl or allvl or benzyl or substituted benzyl, to provide a compound of Formula Illh Formula I or Formula II -tr, o q 11 13 —op(or12)2 (Illh) wherein T, R, and q are as defined for a compound of Formula Illd and R12 is as defined for a compound of Formula X, with the understanding that where q is 2 the compound of Formula Illh will be isolated as a salt with a counterion sufficient to balance the overall charge, and in a second step reacting a compound of Formula Illh with a reagent suitable for deprotecting the phosphate ester, which reagent may be an acid or a base.
Preferably the first step is achieved using a compound of Formula X wherein R11 represents chloride and R12 represents tert-butjd.
Preferably when T is O or T is N and q is 1 the first step is achieved using a base such as sodium hydride or potassium carbonate.
Preferably when T is N and q is 2 the first step is achieved using a polar aprotic solvent such as acetonitrile.
Preferably when R12 represents tert-butyl the second step is achieved using an acid such as trifluoroacetic acid.
Compounds of Formula Illh may also be prepared by reacting alcohol or secondary amine or tertiary amine derivatives of R' or R^ or Z or Formula IX with a compound of Formula X as described, and incorporating the so-formed suitably protected phosphonooxymethyl compound into the general reactions as described above, to provide an alternative route to compounds of Formula Illh.
Received at IPONZ on 14 December 2010 23 Scheme A . i— 100: R = C02H 1 ^ 101: R = NHBoc NHBOC 102 103 i- 104: R = H.HCI v 105: R = COCFi vi CI02S NCOCFf 106 vii cio2s ncocfg 107 NH; 107 viii H0(H2C)2HN02S no, 108 ix NHCbz 109: R = H 110: R = TBDMS i TBDMSOv^^. 111 NH? 107 TBDMS0(H2C)2HN02S 112 no2 107 XJI ROv NHCbz r- 109: R = H ^ 113: R = P(0)(0t-Bu)2 O {t-BuO)2POx^^ 114 NH, (t-Bu0)2(0)P0(H2C)2HN02S xi 115 © (ill) DPPA, Et3N, /-BuOH; 99%. NBS; 100%. 1,3-dichloropropeiie, K,CO-,; 100%.
Received at IPONZ on 14 December 2010 24 Bu-SnH, AIBN then HC1, McOH; 96%.
TFAA, Et-N; 100%.
CISO^H, CH,C1, then (COC1),, DMF; 97%. KNO-, H2S04; 73%.
Ethanolamine, 0 °C then Cs2CO-„ MeOH; 90%. TBDMSC1, DIPEA; 100%.
H„ Pd/C.
DIPEA then Cs2CO„ MeOH; 100% for 112, 95% for 115.
Received at IPONZ on 14 December 2010 Scheme B H0(H2C)2HN02S ... I— 1: R=NO? vm U. 10. r=NH2 vii 115 R0(H2C)2HN02S ix r 123: R=P(0)(0t-Bu)2 ^ 15: R=P(0)(0H)2 (i) 4-(2-Chloroethyl)niorpholine.HCl, K,CCL; 90%. (ii) H2, Pd/C; 99%. (in) NaN02, HCl then MeC0CH(Me)C02Et, NaOAc; 86%. (iv) H2, Pd/C then 4-(2-hydroxyethyl)morpholine, PPhl5 DEAD; 44%. (v) KOH; 96%. (vi) HCl; 91%. (vii) ED CI. HCl, TsOH; 100% for 108, 66% for 123. (viii) H2j PtO,; 74%. (ix) TFA; 98%. 26 Received at IPONZ on 14 December 2010 Scheme C // OBn Et02C H 120 Et02C O, 124 O ho,c 125 115 O -N .HCf H0(H2C)2HN02S ivr 2: R = N02 IV ^ 11: R = NH2 R0(H2C)2HN02S NO, i— 126: R = P(0)(0t-Bu)2 v " 16: R = P(0)(0H)2 (i) H,, Pd/C then 4-(3-hyckoxypropyl)inoipholine, PPh,, DEAD; 47%. (ii) KOH then HCl, 99%'. (iii) EDCI.HC1, TsOH; 100% for 2, 66% for 126. (iv) H,, PtO,; 76%. (v) TFA; 100% Received at IPONZ on 14 December 2010 27 ho2c 129 vi 115 n .hcl O R0(H2C)2HN02S R0(H2C)2HN02S . |— 130; r = tbdms, x = n02 lv 3: r = h, x = n02 12: r = h, x = nh2 vii j— 131: R = P(0){0t-Bu)2 17: R^P(0){0H)2 (i) 4-(2-chloiioethyl)morpholine, NaH; /5%. (ii) ICOH then HCl; 83%. (iii) SOCl2, DMF then 112, DIPEA; 79%. (iv) HCl; 99%. (v) Zn, NH4C1 then HCl; 44%. (vi) EDCI.HC1, TsOH; 94%. (vii) TFA; 80%.
Received at IPONZ on 14 December 2010 28 Scheme E ho2c o.
.HCl ,NMe 133 in 115 R0(H2C)2HN02S NMe . ,— 134: R = TBDMS ivL*4: R = h CI R0(H2C)2HN02S O -NMe no, r— 135: R = P(0)(0t-Bu}2 18: R = P(0)(0H)2 (i) NaH, 1,2-dibromopropane then l-methylpiperazinethen HCl; 48%. (ii) KOH then HCl; 60%. (iii) EDCI, TsOH; 85% for 134, 93% for 135. (iv) HCl; 100%. (v) TFA; 87%.
Received at IPONZ on 14 December 2010 29 Scheme F N02 139 (l) pyBOP, DIPEA then Cs,CO,; 88% for 137, 50% for 139. (ii) EDCI.EI CI, TsOH; 97% for 138, 99% for 140. (in) HCl; 93%. (iv) H2; PtO, then HCl; 89%. (v) HCl, CH,C1,; 85%.
Received at IPONZ on 14 December 2010 Scheme G r 142: R = P(0)(0t-Bu)2 20: R = P(0)(0H)2 © (ii) (m) EDCI.HCl, TsOH; 97% for 141, 81% for 142.
HCl; 100%.
HCl, CH,C1,; 86%. 31 Received at IPONZ on 14 December 2010 Scheme H NO, 144 145 no2 no; 146 147 NO 2 CI ""r^\ T8DMSO(H2C)2HN02S NH iv CI "PL.
H0(H2C)2HN02S 149 NO, NO 2 148 (i) 114, DIPEA, 0 °C; 95%. (ii) Chii-al preparative HPLC (Daicel Chiralpak IA column, EtOH:hexane 40:60). (iii) Cs2C03) MeOH; 100% for 146 and 147. (iv) TFA then acid phosphatase; 91%. (v) TBDMSC1, DIPEA; 100%.
Received at IPONZ on 14 December 2010 Scheme I HO2C- \ H 122 147 N .HCl O R0(H2C)2HN02S II |— 150: R = TBDMS, X = N02 ... n7: R= H, X = N02 14: R = H, X = NH2 R0(H2C)2HN02S . p 151: R = P(0)(0t-Bu)2 21: R=P(OHOI-n, (i) EDCI.HCl, TsOH; 95% for 150 and 151. (ii) HCl, 85%. (iii) Zn, NH4C1; 81%. (iv) TFA; 97%. 33 Received at IPONZ on 14 December 2010 Scheme J HOpC .HCl O 129 147 RO(H2C)2HN02S NO, y [— 152: R = P(0)(0t-Bu)2 22; R = P(0)(0H)2 EDCI.HCl, TsOH; 93%. TFA; 71%.
Received at IPONZ on 14 December 2010 Scheme K VI 158 159 HO^, NH, R0(H2C)2HN02S no2 160: R = H 161: R = TBDMS viii rr ?\ (t-BuO)2PO.^^] 114 NH, (t-Bu0)2(0)P0(H2C)2HN02S IX 162 (i) Nal, 2-butanone; 76%. (ii) Ag0S02Me, CH,CN; 95%. (iii) LiBr, THF; 95%." (iv) LiBr, 2-butanone; 55%. (v) C1S03H then (COCI)2, DMF; 94% for 156, 93% for 157. (vi) KN03j H2S04; 75%. (vii) Ethanolamine then Cs2C03, MeOH; 100%. (viii) TBDMSC1, DIPEA; 95%. (ix) 114, DIPEA then Cs2C03, MeOH; 81%.
Received at IPONZ on 14 December 2010 Scheme L H02C' \ H N .HCl 122 ho2C"\ H 162 N .HBr O 163 H0(H2C)2HN02S R0(H2C)2HN02S 160 i— 164: R=P(0)(0t-Bu)2 23: R=P(0)(0H)2 © © (m) Biorad AG 1-X4 ion exchange resin; 78%. EDCI.HBr, TsOH; 57% for 8, 38% for 164. TFA; 100%. 3g Received at IPONZ on 14 December 2010 Scheme M ho2c N .HCl O 129 H02C 162 .HBr O 165 H0(H2C)2HN02S R0(H2C)2HN02S 160 p 166: R=P(0)(0t-Bu)2 ^ 24: R=P(0)(0H)2 © © (iii) Biorad AG 1-X4 ion exchange resin; 88%. EDCI.HBr, TsOH; 57% for 9, 50% for 166. TFA; 100%. 37 Received at IPONZ on 14 December 2010 Uses of the compounds of the invention The compounds of the Formulae I, II and III of the present invention can be used in the treatment of cancer of the human or animal body. In particular embodiments, the treatment may be of any cancer 5 type that includes hypoxic regions. For example, the cancers treated may be solid tumors, such as ovarian, colon, brain, thyroid, pancreas, bladder, breast, prostate, lung (such as small cell lung tumor cells and large cell lung carcinoma), cervical and skin cancer. Alternatively, the cancer may be leukemia, multiple myeloma or lymphoma.
The compounds of the invention can be administered in the form of pharmaceutical compositions, containing one or more compounds of the invention in combination with one or more pharmaceutically acceptable carriers.
The pharmaceutically acceptable carrier(s) should be non-toxic and not interfere with the efficacy of the 15 active ingredient. The precise nature of the carrier will depend on the route of administration, which can be oral, or parenteral, including intravenous, cutaneous, subcutaneous, intramuscular, intravascular or by infusion.
Pharmaceutical compositions suitable for oral administration can be in tablet, capsule, powder or liquid 20 form. A tablet may comprise one or more solid carriers and/or adjuvants. A capsule may include a solid carrier such as gelatin. liquid pharmaceutical compositions may comprise a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, dextrose or other saccharide solutions or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included.
For parenteral injection, the pharmaceutical composition may conveniently be in the form of a parenterally acceptable aqueous solution which is pyrogen-free and has a suitable pH, isotonicity and stability. Those of skill in the art are able to prepare suitable solutions using, for example, isotonic vehicles such as sodium chloride injection, Ringer's injection, and Lactated Ringer's injection. 30 Preservatives, stabilisers, buffers, antioxidants and/or other additives may be included as required.
The exact dose of the compound to be administered will be at the discretion of the physician, taking into account the condition and needs of the patient. Typical doses and administration schedules will be determined by experience in clinical trials. Total doses are expected to be in the range from about 0.1 35 to 200 mg/kg per subject, such as about 10 mg/kg per subject.
Received at IPONZ on 14 December 2010 The compounds of Formula I, II or III can be used as single agents or in combination with one or more other cytotoxic or other therapeutic agents or therapies, especially those that are relatively ineffective against hypoxic cells, such as radiation therapy. Where such other agents and/or 5 radiotherapy are administered in combination with a compound of the invention, the radiation and/or other agents may be administered before, during or after administration of the compound of Formula I, II or III.
Without wishing to be bound by theory, it is believed that compounds of Formula III, upon 10 administration to a subject, are hydrolysed in vivo to form compounds of Formula I, and that compounds of Formula I are reduced selectively under hypoxic conditions to form cytotoxic amino compounds of Formula II.
EXAMPLES The following examples are representative of the invention, and provide detailed methods for preparing the compounds of the invention. In these examples, elemental analyses were carried out in the Microchemical Laboratory, University of Otago, Dunedin, NZ. Melting points were determined on an 20 Electrothermal 2300 Melting Point Apparatus. NMR spectra were obtained on a Bruker Avance-400 spectrometer at 400 MHz for 1H and 100 MHz for 13C spectra, referenced to Me4Si. Column chromatography was carried out on silica gel unless otherwise stated.
Example 1. T(Chloromethyl)-iV(2-hydroxyethyl)-3-[5-(2-morpholinoetlioxy)-U:/-indole-2-25 carbonyl]-5-nitro-i,2-dihydro-3//-benzo[e]indole-7-sulfonamide (1) (Scheme A and B). Et,N (89.2 mL, 638 mmol) was added to solid 2-naphthoic acid 100 (100 g, 581 mmol) and the mixture was stirred at 20 °C for 10 min. After this time the mixture solidified and was allowed to stand at 20 °C for a further 20 min. Diphenyl phosphoryl azide (131.6 mL, 609 mmol) was added, followed by dry ftr/-BuOH (277.2 mL, 2.90 moles), causing the internal temperature to rise to 35 °C. The 30 reaction flask was placed in an oil-bath and the bath temperature was raised from 20 °C to 86-87 °C over 1 h 20 min, then held at this temperature for 2 h. The reaction mixture was cooled briefly and poured into ice-water (ca.2.5 L) with vigorous stirring at 20 °C causing a sticky solid mass to separate. The mixture was left to stand at 20 °C overnight (15 h) to give a loose suspended solid. The mixture was stirred again for 5 h. The fine solid was filtered off, washed with water (4 x 300 3 5 mL) and dried in vacuum oven at 40-45 °C to constant weight to give terl-bwvA 2-naphthylcarbamate 39 Received at IPONZ on 14 December 2010 (101) as a beige solid (140 g, 99%): mp 85-87 °C, suitable ('H NMR) for the following reactions. A portion was recrvstallised from CH2CI2/petroleum ether to give a cream solid: 'H NMR (CDC13) 8 7.98 (s, 1 H), 7.75 (d J = 8.6 Hz, 3 H), 7.43 (dtj = 7.5, 1.1 Hz, 1 FI), 7.40-7.30 (m, 2 H), 6.61 (s, 1 H), 1.55 (s, 9H).
A stirred solution of carbamate 101 (92 g, 0.38 mol) in MeCN (680 mL) at 0 DC was treated portionwise over 2 li with solid A-bromosuccinimide (81 g, 0.45 mol). After the addition was complete the reaction mixture was stirred for a further 45 min at 0 °C. To the mixture was added a cold solution of aqueous NaHCOs (0.2N, 2 L) and the mixture was stirred at 0 °C for 1 h. The solid 10 was filtered off and washed with water (5 X 200 mL), then dried in vacuum over solid KOH overnight to give /erZ-butyl l-bromo-2-naphthylcarbamate (102) as a beige solid (121 g, 100%): mp 88-89 °C; aH NMR [(CD3)2SO] 5 8.82 (s, 1 H), 8.15 (dJ = 8.5 Hz, 1 H), 7.96 (d,J= 9.6 Hz, 1 H), 7.93 (d J = 9.3 Hz, 1 H), 7.71 (dJ = 8.8 Hz, 1 H), 7.66 (t J - 7.7 Hz, 1 H), 7.56 (t J = 7.4 Hz, 1 H), 1.49 (s, 9 H).
A mixture of carbamate 102 (53.7 g, 167 mmol) and dry K2C03 (55.2 g, 400 mmol) in dry DMF (500 mL) was stirred at 20 °C for 5 min, then 1,3-dichloropropene (23.1 mL, 251 mmol, mixed isomers) was added. The mixture was stirred at 70-80 °C (bath temperature) under a dry atmosphere for 4 h. The mixture was cooled and partitioned between petroleum ether (500 mL) and water (500 mL). 20 The petroleum ether layer was separated and washed with water (3 x 200mL), then dried (Na2S04) and allowed to stand at 5 °C overnight. The precipitated orange solid was filtered off and the filtrate was evaporated to give tert-butyl l-bromo-2-naphthyl-(3-chloro-2-propen-l-yl)carbamate (103) as an amber oil (67.5 g; 102%), suitable ('FI NMR) for the following reactions. Pure carbamate 103 can be obtained by repeating the procedure of dissolving in petroleum ether, allowing an orange solid to 25 separate, filtering off the solid, and evaporating the filtrate, to give 103 as an oil (100%): 111 NMR [(CD3)2SO] (mixture of rotamers and E and Z forms) 5 8.23 (d,J = 8.4 Hz, 1 H), 8.07-7.94 (m, 2 H), 7.71 (t ,J= 7.5 Hz, 1 H), 7.65 (tJ = 7.4 Hz, 1 H), 7.51, 7.45 (2 dj = 8.6 Hz, 1 H), 6.44-6.26 (m, 1 H), 6.21-5.99 (m, 1 FI), 4.58^4.46, 4.44-4.17, 4.14-3.96 (3 m, 2 H), 1.50, 1.26 (2 s, 9 H).
A solution of bromide 103 (48.8 g, 123 mmol) in toluene (150 mL) was stirred at 20 °C under nitrogen for 20 min, then Bu3SnH (34.2 mL, 123 mmol) and AIBN (60.7 mg, 0.37 mmol) were added. The reaction mixture was stirred at 85-90 °C (bath temperature) under nitrogen for 1 h and then cooled to 20 °C. Powdered anhydrous KF (35.7 g, 615 mmol) was added and the mixture was stirred at 20 °C for 14 h. EtOAc (150 mL) and petroleum ether (300 mL) were added and the 35 mixture stirred for a further 1 h 30 min. The mixture was filtered through a neutral alumina pad (300 40 Received at IPONZ on 14 December 2010 g), washing with EtOAc/petroleum ether (1:2) (10 x 50 mL). The combined filtrates were washed successively with water (300 mL), aqueous Na2CO, (5%, 300 mL) and water (300 mL), and then dried (Na2S04) and evaporated to give a yellow solid (74 g). 'H NMR analysis showed this material contained about 10% tin residues. The solid was dissolved in MeOH (150 mL) and HCl gas was 5 bubbled into the solution at room temperature over 25 min. The acidic mixture was stirred for a further 10 min, then diluted with petroleum ether (300 mL). The mixture was stirred at 20 °C for 15 min, then the petroleum ether layer was separated and discarded. The petroleum ether procedure was repeated three more times. The acidic MeOH solution was then evaporated under reduced pressure at 35-40 °C (bath temperature) to give l-(chloromethyl)-l,2-dihydro-3H-benzo[Vjindole 10 hydrochloride (104) as a greenish foamy solid (30.1 g, 96%): mp 180 °C (dec.); 1H NMR [(CD3)2SO] 8 10.3 (v br, 1 H), 8.07-7.94 (m, 3 H), 7.67-7.59 (m, 1 H), 7.57-7.49 (m, 1 H), 7.45 (d ,J= 8.7 Hz, 1 H), 4.45-4.36 (m, 1 H), 4.08 (dd,/= 11.0, 3.5 Hz, 1 H), 4.00-3.90 (m, 2 H), 3.84 (dd,/= 11.5, 2.5 Hz, 1 H). This material contained about 1% tin residues. On a smaller scale (3.0 g) repeated washes with petroleum ether (4 x 100 mL) were sufficient to remove all traces of tin residues to give an 15 analytically pure product: Anal. (Ct3H13Cl2N'0.4H20) Calc: C, 59.74; H, 5.32; N, 5.36. Found: C, 59.71; H, 5.36; N, 5.32.
To a stirred mixture of indoline hydrochloride 104 (30.1 g, 118 mmol) in p-dioxane (100 mL) at 0 °C was added trifluoroacetic anhydride (33.4 mL, 236 mmol), followed by Et3N (49.2 mL, 354 mmol). 20 The mixture was stirred at 0 °C for 10 min, at 20 °C for another 50 min, and then poured into ice-water (500 mL). Cone. HCl (40 mL) was added and the mixture was stirred at 20 °C for 20 min. The precipitated solid was filtered off, washed successively with dilute HCl (IN, 4 x 20 mL), water (10 x 50mL), petroleum ether (5 x 25 mL), and dried to give l-(chloromethyl)-3-(trifIuoroacetyl)-l,2-dihydro-3H-benz]>]indole (105) as a pale yellow solid (37.7 g, 102%) suitable for the following 25 reactions: NMR [(CD3)2SO] 8 8.32 (d J = 9.0 Hz, 1 FI), 8.07-7.96 (m, 3 H), 7.62 (dddj = 8.2, 6.9, 1.2 FIz, 1 FI), 7.53 (dddj - 8.1, 6.9, 1.1 Hz, 1 H), 4.61-4.52 (m, 1 H), 4.51-4.39 (m, 2 H), 4.15 (dd, J — 11.3, 3.0 Hz, 1 H), 4.04 (dd,J— 11.3, 5.9 Hz, 1 FI). This material was free of tin residues NMR) but contained a small impurity of Et3N that can be removed by further washing with FIC1.
A solution of chlorosulfonic acid (1.77 mL, 26.5 mmol) in dry CFI2C12 (20 mL) at 20 °C was added dropwise over 1 h to a stirred mixture of indoline 105 (6.92 g, 22.1 mmol) and CH2C12 (60 mL) in an ice bath, maintaining the internal temperature at 3-4 °C throughout. The mixture was stirred for a further 2 h 30 min at the same temperature. Another batch of chlorosulfonic acid (1.77 mL, 26.5 mmol) in dry CH2C12 (10 mL) was added dropwise over 45 min and the reaction was stirred in the Received at IPONZ on 14 December 2010 ice bath for a further 1 h 30 min. Dry DMF (10 mL) was added followed by oxalyl chloride (5.55 mL, 63.6 mmol). The reaction mixture was stirred in the ice bath for 30 min and then left to stand at 0-5 °C overnight. The CH2C12 was evaporated under reduced pressure at 20 °C to give an amber solid. Cold water (100 mL) was added and the mixture was stirred at 0 °C (bath temperature) for 15 5 min. The solid was filtered off, washed with cold water (5 x 20 mL), and dried to give 1- (chloromethyl)-3-(trifluoroacetyl)-l,2-dihydro-3H-benzo[>]indole-7-sulfonyl chloride (106) as a pale yellow solid (8.8 g, 97%): mp 187-190 °C; NMR (CDC1,) 5 8.70-8.64 (m, 2 H), 8.13-8.08 (m, 2 H), 8.00 (d J = 9.0 Hz, 1 H), 4.72-4.68 (m, 1 H), 4.55-4.48 (m, 1 FI), 4.33-4.25 (m, 1 H), 3.98-3.93 (m, 1 H), 3.68-3.61 (m, 1 H). The 'H NMR spectrum was identical to that of a sample recrystallised 10 from EtOAc/petroleum ether, mp 189-192 °C.
Sulfonyl chloride 106 (1.02 g, 2.48 mmol) was added to cold conc. H2S04 (10 mL) and the mixture was stirred at 0 °C for 2 min to give a homogeneous solution. Solid KN03 (351 g, 3.47 mmol) was added portionwise over 2 min and the mixture was stirred at 0 °C for a farther 2 min. Ice-water (100 15 mL) was added and the mixture was stirred at 0 °C for 2 min. The precipitated solid was filtered off, washed with cold water (5 x 20mL), and dried in vacuum over silica gel to give l-(chloromethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3H-benzo[^]indole-7-sulfonyl chloride 107 as a light brown solid (1.09 g, 97%) suitable ('Ft NMR) for the following reactions. The product can be further purified by dissolving in CH2C12, diluting with i-Pr20 (2 volumes), allowing the solution to stand at 5 °C overnight, and filtering off the precipitate as a light brown solid (71%). The mother liquor left at -10 °C gives an additional crop (0.48 g, 2%). A recrystallised sample has mp (EtOAc/petroleum ether) 184-189 °C; 'H NMR (CDC13) S 9.34 (s, 1 H), 9.28 (d J = 1.8 Hz, 1 H), 8.22 (dd J = 9.0, 1.9 Hz, 1 H), 8.11 (d J = 9.0 Hz, 1 H), 4.77-4.71 (m, 1 H), 4.58 (ddj =11.5, 8.8 Hz, 1 H), 4.42-4.33 (m, 1 H), 3.95 (dd J = 25 11 .7, 3.5 Hz, 1 H), 3.73 (dd, J = 11.7, 7.7 Hz, 1 FI).
Sulfonyl chloride 107 was converted to alcohol 108 as previously described (formation of compound 121 of WO 2006/043839).
To a mixture of benzyl 2-hydroxyethylcarbamate (109) (3.90 g, 20.0 mmol) and lert- butylchlorodimethylsilane (4.52 g, 30.0 mmol) in DMF (50 mL) stirred at 20 °C was added di-iso-propylethylamine (DIPEA) (12.0 mL, 68.8 mmol). The reaction mixture was stirred for a further 20 min and then partitioned between EtOAc and water. The EtOAc layer was separated, washed with dilute aqueous Na2C03, dried (Na2S04), and evaporated to give benzyl 2-itert-35 butyldimethylsilyloxy)ethylcarbamate (110) as a pale amber oil (6.2 g, 100%):111 NMR (CDC13) 5 Received at IPONZ on 14 December 2010 7.38-7.28 (m, 5 H), 5.11 (s, 2 H), 5.06 (br s, 1 H), 3.68 (tJ = 5.3 Hz, 2 H), 3.31 (q,/= 5.3 Hz, 2 H), 0.89 (s, 9 H), 0.05 (s, 6 H). HRMS (FAB) calcd. for C16H28N03Si (MH+) 310.1838, found 310.1838.
A solution of carbamate 110 (1.5 g, 4.85 mmol) in THF (100 mL) was hydrogenated over Pd/C (5%) at 40 psi for 2 h 30 min. The mixture was filtered through Celite and the Celite was washed with THF (5 X 20 mL). To the filtrate containing amine 111 was added sulfonyl chloride 107 (1.84 g, 4.03 mmol) and DIPEA (0.84 mL, 4.85 mmol). The mixture was stirred at 0 °C for 40 min. Cesium carbonate (1.31 g, 4.03 mmol) and MeOH (50 mL) were added and the mixture was stirred at 20 °C 10 for 45 min, then allowed to stand at 5 °C for 14 h. The mixture was concentrated under reduced pressure at 25 °C to remove most of the THF. Ice-water was added and stirred at 0 °C for lh 30 min. The resulting solid was filtered off, washed with cold water several times, and dried in vacuum over silica-gel to give IV-[2-(^-butyldimethylsilyloxy)ethyl]-l-(chloromethyl)-5-nitro-l,2-dihydro-3ii-benzo[c]indole-7-sulfonamide (112) as a reddish orange solid (2.0 g, 100%): mp 127-129 °C; 111 15 NMR (CDClj) 8 8.92 (d J = 1.6 Hz, 1 H), 7.90 (ddJ = 8.9, 1.6 Hz, 1 H), 7.79 (d J = 8.9 Hz, 1 H), 7.73 (s, 1 H), 4.88 (t,J = 5.8 Hz, 1 H), 4.40 (br s, 1 H), 4.17-4.08 (m, 1 FI), 4.05-3.95 (m, 2 H), 3.83- 3.74 (m, 1 H), 3.68 (tJ = 5.2 Hz, 2 H), 3.60 (ddj = 11.1, 10.0 Hz, 1 H), 3.18-3.11 (m, 2 H), 0.84 (s, 9 H), 0.00 (s, 6 H). Anal. (C21H30ClN3O5SSi) Calc: C, 50.44; H, 6.05; N, 8.40. Found: C, 50.71; FI, 6.35; N, 8.32.
Tetrazole (3 wt% solution in CH3CN, 468 mL, 159 mmol) was added gradually over 1 h to a stirred mixture of di-W-butyl N,JV-diisopropylphosphoramidite (95%, 53.0 mL, 159 mmol) and benzyl 2-hydroxyethylcarbamate (109) (26.0 g, 133 mmol) in THF (500 mL) at 20 °C under a nitrogen atmosphere. After the addition was complete the mixture was stirred at this temperature for 19 h. 25 The mixture was cooled to 0 °C and H202 (30% aqueous, 48 mL, 494 mmol) was added. After 1.5 h at 0 °C the mixture was poured into ice-water and the product was extracted into EtOAc. The EtOAc layer was washed successively with cold aqueous Na2C03 and water (x 2) and then dried (Na2S04). The organic solvents were removed under reduced pressure to give a colourless oil. A seed crystal was added and the mixture was allowed to stand at 20 °C for 1 h, then petroleum ether 30 was added and the mixture was left at 5 °C for crystallisation to complete. The resulting solid was filtered off, washed with petroleum ether, and dried to give benzyl 2- {[di(/tr/-butoxy)phosphoryl]oxy}ethylcarbamate (113) (24.9 g, 49%) as a colourless solid: mp 43-45 °C; 'H NMR (CDClj) 8 7.37-7.28 (m, 5 H), 5.43 (br s, 1 H), 5.11 (s, 2 FI), 4.07-4.00 (m, 2 H), 3.46 (qJ = 5.1 Hz, 2 H), 1.47 (s, 18 H). HRMS (FAB) calc for C18H31N06P (MH+) ml^ 388.1889, found 35 388,1889. The mother liquor was left at 5 °C to give a second crop (12.9 g, 25%).
Received at IPONZ on 14 December 2010 A solution of benzyl carbamate 113 (2.32 g, 5.99 mmol) in MeOH (100 mL) with Pd/C (5%, 0.46 g) was hydrogenated at 40 psi for 2.5 h. The mixture was filtered through Celite, washing with MeOH, and the filtrate was evaporated to give 2-aminoethyl di(^-butyl) phosphate (114) (1.52 g, 100%) as a 5 colourless oil: *H NMR (CDC1,) S 4.01-3.94 (m, 2 H), 2.96-2.90 (m, 2 H), 1.58 (br s, 2 H), 1.49 (s, 18 H). HRMS (FAB) calc for C10H25NO4P (MH+) w/? 254.1521, found 254.1519.
A cold solution of amine 114 (1.98g, 7.8 mmol) and DIPEA (1.22 mL, 6.8 mmol) in THF (10 mL) was added to solution of sulfonyl chloride 107 (2.98 g, 6.5 mmol) in THF (30 mL) at 0 °C. The 10 mixture was stirred at 0 °C for a further 20 min, then solid Cs2CO, (4.04 g, 12.4 mmol) and cold MeOH (20 mL) were added. The mixture was stirred at 0 °C for a further 10 min and then partitioned between EtOAc (400 mL) and ice-water (400 mL). The organic layer was separated and the aqueous layer was further extracted with EtOAc. The combined EtOAc extracts were washed again with water, then dried (Na2S04), and evaporated under reduced pressure at 25 °C (bath 15 temperature). The residue was dissolved in CH2C12 and the solution was filtered through a short silica column, eluting with CH2CI2. The product-containing fractions were evaporated to give di[tert-butyl) 2-[l-(chloromethyl)-5-nitro-l,2-dihydro-3H-benzoj>]indole-7-sulfonamido]ethyl phosphate (115) as a red-orange foam (3.60 g, 95%): 'H NMR [(CD3)2SO] 5 8.59 (d,]- 1.6 Hz, 1 H), 8.04 (dJ - 8.9 Hz, 1 H), 7.95 (br s, 1 H), 7.79 (ddj = 8.9, 1.8 Hz, 1 H), 7.77 (s, 1 H), 6.74 (s, 1 H), 4.28-4.20 20 (m, 1 H), 3.95-3.86 (m, 2 H), 3.83-3.72 (m, 4 H), 2.99 (t J = 6.0 Hz, 2 H), 1.35 (s, 18 H). HRMS (FAB) calc for C^H^CINjOgPS (M+) m/% 577.1415, found 577.1412.
To a stirred mixture of 4-nitrophenol (116) (42 g, 0.30 mol) and 4 (2- ch 1 oroethy 1)morp h o 1 i ne hydrochloride (56 g, 0.30 mol) in anhydrous DMF (400 mL) was added dry K2CO, (104 g, 0.75 25 mol). The mixture was stirred under a nitrogen atmosphere at 80-90 °C (bath temperature) for 3 h, then at 20 °C overnight (15 h). The mixture was poured into ice-water (1.5 L) and stirred at 0 °C for 1 h. The precipitated solid was filtered off, washed with water (6 x 100 mL) until the washes were colourless, then dried to give 4-[2-(4-nitrophenoxy)ethyl]morpholine (117) as a pale yellow solid (68.4 g, 90%): mp 79-81 °C; 1H NMR (CDCl,) 8 8.24-8.18 (m, 2 H), 7.01-6.93 (m, 2 H), 4.20 (t J = 30 5.7 Hz, 2 H), 3.73 (t, J = 4.7 Hz, 4 H), 2.84 (t, J = 5.7 Hz, 2 H), 2.58 (t, J = 4.7 Hz, 4 H). Anal. (C12H1GN204) Calc: C, 57.13; H, 6.39; N, 11.10. Found: C, 57.36; H, 6.44; N, 11.25.
A solution of nitro compound 117 (33.3 g, 132 mmol) in MeOH (170 mL) and THF (50 mL) with Pd/C (5%, ca. 3 g) was hydrogenated at 40 psi overnight (17 h). The mixture was filtered through 35 Celite and the Celite plug was washed with CH2Cl2:MeOH (5:1) several times. The combined Received at IPONZ on 14 December 2010 filtrates were evaporated to give 4-(2-morphoUnoethoxy)aniline (118) as a pink solid (29.0 g, 99%): mp 39-40 °C; ?H NMR (CDC1,) 5 6.78-6.73 (m, 2 H), 6.66-6.61 (m, 2 H), 4.04 (t ,J~ 5.8 Hz, 2 H), 3.73 (t4.7 Hz, 4 H), 3.41 (br s, 2 H), 2.76 (t J - 5.8 Hz, 2 H), 2.57 (t J - 4.7Hz, 4 H). Anal. (C12H18N202) Calc: C, 64.84; H, 8.16; N, 12.60. Found: C, 64.94; H, 8.08; N, 12.66.
A solution of sodium nitrite (9.95 g, 144 mmol) in water (27 mL) was added to a mixture of aniline 118 (29.0 g, 131 mmol), water (223 mL) and conc. HCl (65.7 mL, 657 mmol) at 0 °C. The mixture was stirred for a further 10 min at 0 °C, then added to a stirred mixture of ethyl 2-methyl-3-oxobutanoate (19.8 g, 138 mmol), sodium acetate (112 g, 1.37 mol), EtOH (163 mL), and ice (133 g, 10 freshly added just before mixing) at 0 °C (bath temperature). The final reaction mixture was stirred at 0 °C for 5 min, then at 20 °C for 1 h 40 min. Solid Na2C03 was added to give a pH of approximately 7-8. The mixture was extracted with CH2C12 (2 x 300 mL) and the extracts were washed with cold water (400 mL), then dried (Na2S04) and evaporated to give a red oil. To the oil was added EtOH (60 mL) and HCl-saturated EtOH (100 mL). The mixture was stirred at reflux for 15 30 min, then cooled and evaporated under reduced pressure to give a dark solid. Water (200 mL) was added and the aqueous mixture was basified at 0 °C with 10% aqueous Na2C03 solution to a pFI of approximately 7-8. The resulting pale orange solid was filtered off, washed with water (5 x 80 mL), and dried to give ethyl 5 (2 morpholinocfhoxyj 1indole 2 carboxylate (119) as a yellow solid (35.9 g, 86%): mp 130-132 °C; 'FI NMR [(CD3)2SO] 8 11.69 (s, 1 FI), 7.34 (d J = 9.0 Hz, 1 H), 7.12 20 (dj = 2.3 Hz, 1 H), 7.03 (s, 1 H), 6.91 (ddJ = 8.9, 2.5 Hz, 1 H), 4.32 (qJ - 7.1 Hz, 2 H), 4.07 (t,J = 5.8 Hz, 2 FI), 3.61-3.56 (m, 4 H), 2.71 (t,J ~ 5.8 Hz, 2 H), 2.50-2.46 (m, 4 H, partially obscured by DMSO peak), 1.33 (t J = 7.1 FIz, 3 FI). Anal. (C17H22N204) Calc: C, 64.14; H, 6.97; N, 8.80. Found: C, 63.82; H, 7.23; N, 8.91.
Pd/C (5%) was added to a solution of ethyl 5-(benzyloxy)-lH-indole-2-carboxylate (120) (5.10 g, 17.3 mmol) in EtOAc (40 mL), THF (40 mL), and EtOH (30 mL). The mixture was hydrogenated at 50 psi for 17 h, then filtered through Celite. The filtrate was evaporated to give crude ethyl 5-hydroxy-lH-indole-2-carboxylate as a grey solid. A portion of this crude material (5.8 mmol) was dissolved in THF (40 mL) with triphenylphosphine (2.26 g, 8.7 mmol) and 4-(2-30 hydroxyethyl)morpholine (1.06 mL, 8.7 mmol). Diethyl azodicarboxylate (1.50 g, 8.7 mmol) was added dropwise and the mixture was stirred at room temperature for 2 days. The THF was evaporated and the residue was partitioned between EtOAc and water. The EtOAc layer was washed with water (X2) and extracted with aqueous HCl (2N, X2). The combined acid extracts were cooled to 0 °C, basified with conc. aqueous NH3, and extracted with EtOAc (x2). The combined 35 organic extracts were dried (MgS04) and evaporated to give a light brown oil which crystallised on 45 Received at IPONZ on 14 December 2010 standing. Recrystallisation from EtOH gave indole 119 as pale yellow crystals (0.80 g, 44%): mp 130-132 °C; 'H NMR [(CD3)2SO] identical to that described above.
KOH (19.0 g, 339 mmol) was added to a mixture of ester 119 (35.9 g, 113 mmol), MeOH (250 mL) 5 and water (125 mL) and the mixture was stirred at 20 °C for 1 h. The MeOH was evaporated under reduced pressure at 40 °C (bath temperature). The basic aqueous mixture was washed with ether (3 x 200 mL), filtered, and the filtrate acidified with conc. HCl at 0 °C to pH 5-6. The precipitated solid was filtered off, washed with water many times (until the washings were no longer red), and dried in a vacuum oven at 50-60 °C for 7 h to give 5-(2-morpholinoethoxy)-lH-indoIe-2-carboxylic acid 10 (121) (31.3 g, 96%) as a pale yeUow solid; *H NMR [(CD3)2SO] 8 12.80 (br s, 1 H), 11.56 (s, 1 H), 7.32 (dJ = 8.9 Hz, 1 H), 7.12 (dJ = 2.4 Hz, 1 PI), 6.99-6.96 (m, 1 PI), 6.89 (ddj = 8.9, 2.4 Hz, 1 H), 4.07 (t J = 5.8 Hz, 2 H), 3.59 (tj = 4.6 Hz, 4 H), 2.71 (tj = 5.8 Hz, 2 H), (4H partially obscured by DMSO peak).
Dioxane (300 mL) and HCl in dioxane (4M, 40.5 mL, 162 mmol) were added to acid 121 (31.3 g, 108 mmol). The mixture was stirred at 20 °C for 15 h. The solid was filtered off, washed with EtOAc several times, and dried to give 5-(2-morpboHnoethoxy)-l H-indote 2 carboxylic acid hydrochloride (122) as a grey-white solid (32 g, 91%): mp 251-254 °C; 1H NMR [(CD3)2SO] 5 12.86 (br s, 1 H), 11.66 (s, 1 H), 10.97 (br s, 1 H), 7.36 (d J = 8.9 Hz, 1 H), 7.20 (dj= 2.2 Hz, 1 H), 7.01 (dj = 1.4 20 Hz, 1 H), 6.97 (ddj = 8.9, 2.4 Hz, 1 H), 4.44-4.37 (m, 2 EI), 4.03-3.92 (m, 2 H), 3.88-3.75 (m, 2 H), 3.60-3.45 (m, 4 H), 3.26-3.15 (m, 2 H). Anal. (C15FIlt)N204HCl) Calc: C, 55.13; H, 5.86; N, 8.57. Found: C, 55.02; H, 5.89; N, 8.50.
A mixture of indoline 108, acid 122 (104 mg, 0.32 mmol), EDCI.HCl (175 mg, 0.92 mmol), and 25 anhydrous TsOH (7.9 mg, 0.05 mmol) in DMA (2,5 mL) was stirred at room temperature for 2.5 h then cooled in an ice bath. Cold aqueous NaH CO, (5%) was added and the precipitated solid was filtered off, washed with water, and dried to give 1 as a yellow solid (150 mg, 100%): mp 131-135 °C; XH NMR [(CD3)2SO] 5 11.73 (dj = 1.7 Hz, 1 H), 9.29 (s, 1 H), 8.85 (dj = 1.7 Hz, 1 H), 8.43 (dj - 8.9 Hz, 1 H), 8.03 (ddj = 8.9,1.7 Hz, 1 H), 7.91 (t J = 5.6 Hz, 1 H), 7.41 (dJ = 8.9 Hz, 1 H), 30 7.21 (dj = 1.8 Hz, 1 H), 7.19 (dj= 2.3 Hz, 1 H), 6.96 (ddj = 8.9, 2.4 Hz, 1 H), 5.01-4.94 (m, 1 H), 4.73 (ddj = 10.9, 2.4 Hz, 1 H), 4.68-4.62 (m, 1 FI), 4.67 (tJ = 5.6 FIz, 1 H), 4.20-4.10 (m, 4 H), 3.64-3.58 (m, 4 H), 3.39 (qJ = 6.0 Hz, 2 H), 2.87 (qJ = 6.2 Hz, 2 H), 2.73 (tj = 5.7 Hz, 2 H), ca. 2.52-2.46 (m, 4 H, partially obscured by DMSO peak). Anal. (C, ::i H 32 C1N 3 O H S' % 1120) Calc: C, 53.65; H, 5.03; N, 10.43. Found: C, 53.79; H, 4.85; N, 10.37.
Received at IPONZ on 14 December 2010 Example 2. 5-Amino-l-(chloromethyl)~A^(2-hydfoxyethyl)-3- [5-(2-morpholinoethoxy)-l//-indole-2-cafbonyl]-l,2-dihydfo-3i^benzo[e]indole-7-sulfonamide (10) (Scheme B). A solution of nitro compound 1 (49.6 mg, 0.075 mmol) in THF (20 mL) with Pt02 (59 mg) was hydrogenated at 50 psi for 70 min. The catalyst was filtered off through Celite, the filtrate was 5 evaporated, and the residue was triturated with EtOAc to give 10 as a pale green solid (35 mg, 74%): mp 220-225 °C (dec.); *H NMR [(CD3)2SO] 8 11.57 (s, 1 H), 8.54 (d J = 1.6 Hz, 1 H), 7.94 (d J = 8.9 FIz, 1 H), 7.82 (s, 1 H), 7.74 (ddj = 8.8, 1.7 Hz, 1 H), 7.45 (br s, 1 H), 7.40 (dj = 8.9 Hz, 1 H), 7.18 (dj — 2.3 Hz, 1 H), 7.09 (dJ = 1.2 Hz, 1 H), 6.93 (ddJ = 8.9, 2.4 Hz, 1 H), 6.29 (s, 2 H), 4.76 (ddj = 10.8, 9.0 Hz, 1 H), 4.65 (tJ = 5.4 Hz, 1 H), 4.53 (ddj = 10.9, 1.8 Hz, 1 H), 4.21-4.14 (m, 1 10 H), 4.12 (tj = 5.8 Hz, 2 H), 4.01 (ddj - 11.0, 3.1 Hz, 1 H), 3.80 (ddj = 11.0, 7.7 Hz, 1 H), 3.63-3.58 (m, 4 H), 3.40 (qJ - 6.0 Hz, 2 H), 2.84 (br tJ = 6.2 Hz, 2 H), 2.72 (tj = 5.8 Hz, 2 H), ca. 2.52-2.46 (m, 4 H, partially obscured by DMSO peak). Anal. (C30H34ClNsO6S) Calc: C, 57.37; H, 5.46; N, 11.15. Found: C, 57.12; H, 5.36; N, 11.00.
Example 3. 2-{1-(Chloromethyl)-3-[5-(2-moqiholinoethoxy)-l//-indole-2-carboiiyl]-5-iiitro-l,2-dihydro-3/£benzo[e]indole-7-sulfonamido}ethyl dihydrogen phosphate trifluoroacetate (15) (Scheme B). A mixture of indoline 115 (182 mg, 0.31 mmol), acid 122 (134 mg, 0.40 mmol), EDCI.HCl (241 mg, 1.24 mmol), and anhydrous TsOH (11 mg, 0.06 mmol) in DMA (2 mL) was stirred at room temperature for 2 h then cooled in an ice bath. Cold aqueous NaHC03 (5%) was 20 added and the precipitated solid was filtered off, washed with water, and dried. The resulting solid was triturated with acetone to give di-zW-butyl 2-{l-(chloromethyl)-3-[5-(2-morpholinoethoxy)-lH-indole-2-carbonyl]-5-nitro-l,2-dihydro-3H-benzo[^]indole-7-sulfonamido}ethyl phosphate (123) as a yellow solid (176 mg, 66%): mp 200-204 °C (dec.); 'H NMR [(CD3)2SO] S 11.77 (d J = 1.6 Hz, 1 H), 9.30 (s, 1 H), 8.86 (dJ = 1.7 Hz, 1 H), 8.45 (d J = 8.9 Hz, 1 H), 8.20 (t J = 5.9 FIz, 1 H), 8.00 25 (dd, J = 8.9, 1.7 Hz, 1 H), 7.39 (d, J = 8.9 Hz, 1 H), 7.21 (dj -1.7 Hz, 1 H), 7.08 (d, J = 2.3 Hz, 1 H), 6.95 (ddj = 8.9, 2.4 Hz, 1 H), 5.01-4.94 (m, 1 H), 4.71 (ddJ = 10.9, 2.2 Hz, 1 H), 4.69-4.63 (m, 1 H), 4.20-4.09 (m, 4 H), 3.83 (qj = 6.3 Hz, 2 H), 3.63-3.58 (m, 4 H), 3.06 (qJ = 5.8 FIz, 2 H), 2.73 (tj = 5.7 Hz, 2 H), ca. 2.52-2.46 (m, 4 H, partially obscured by DMSO peak), 1.35 (s, 18 H). Anal (C38H49ClN5OnPS) Calc: C, 53.68; H, 5.81; N, 8.24. Found: C, 53.59; H, 5.61; N, 8.19.
TFA (0.15 mL, 1.9 mmol) was added to a suspension of the phosphate ester 123 (164 mg, 0.19 mmol) in CH2C12 (5 mL) and the resulting solution was stirred at room temperature for 16 h, during which time a precipitate separated. The mixture was evaporated, the residue was resuspended in CH2C12 and evaporated once more, and the residue was triturated with EtOAc to give 15 as a yellow 35 solid (158 mg, 98%): mp 166-169 °C; 1H NMR [(CD3)2SO] 5 11.84 (s, 1 H), ca. 10.7 (v br s, 1 H), 47 Received at IPONZ on 14 December 2010 9.29 (s, 1 H), 8.87 (dJ = 1.7 Hz, 1 H), 8.45 (d J = 8.9 Hz, 1 H), 8.27-8.20 (m, 1 H), 8.03 (ddj = 8.9, 1.7 Hz, 1 H), 7.44 (dj = 8.9 Hz, 1 H), 7.26 (dj - 2.2 Hz, 1 H), 7.24 (dj = 1.8 Hz, 1 H), 7.02 (ddj = 8.9, 2.3 Hz, 1 H), 5.01-4.94 (m, 1 H), 4.71 (ddj = 10.8, 2.1 Hz, 1 H), 4.68-4.62 (m, 1 H), 4.34-4.29 (m, 2 H), 4.19-4.09 (m, 2 H), 3.87-3.76 (m, 6 H), 3.26-3.13 (m, 4 H), 3.06-3.00 (m, 2 H), (2 5 H not observed, obscured by water peak 8 3.5-3.3). Anal. (C30H3,ClN5Ol1PS'CF3CO,H) Calc: C, 45.11; H, 4.02; N, 8.22. Found: C, 44.72; H, 4.36; N, 8.57.
Example 4. l-fChloromethy^-iV-(2-hydroxy ethyl)-3- [5-(3-morpholinopropoxy)-l/£indole-2-carbonyl]-5-nitro-l,2-dihydro-3i^benzo[e]indole-7-sulfonamide (2) (Scheme C). Crude ethyl 5-hydroxy-li-f-indole-2-carboxylate as prepared in Example 1 (5.51 mmol) was dissolved in THF (40 mL) with triphenylphosphine (2.17 g, 8.3 mmol) and 4-(3-hydroxypropyl)morpholine (1.20 g, 8.3 mmol). Diethyl azodicarboxylate (1.44 g, 8.3 mmol) was added dropwise and the mixture was stirred at room temperature for 2.5 h. The THF was evaporated and the residue was partitioned between EtOAc and water. The EtOAc layer was washed with brine (x3) and extracted with aqueous HCl (2N, X2). The combined acid extracts were cooled to 0 °C, basified with conc. aqueous NH3, and extracted with EtOAc (x3). The combined organic extracts were dried (MgS04) and evaporated to give a tan solid. Recrystallisation from EtOH gave ethyl 5-(3-morpholinopropoxy)-lH-indole-2-carboxylate (124) as pale yellow crystals (0.86 g, 47%): mp 152-155 °C; 11! NMR [(CD3)2SO] 6 11.68 (s, 1 H), 7.33 (dj = 8.9 Hz, 1 H), 7.10 (dJ = 2.3 FIz, 1 FI), 7.03 (s, 1 H), 6.91 (ddj - 8.9, 2.4 Hz, 1 H), 4.33 (qj = 7.1 Hz, 2 H), 3.99 (t, J = 6.4 Hz, 2 H), 3.60-3.56 (m, 4 H), 2.43 (t J = 7.2 FIz, 2 H), 2.39-2.35 (m, 4 FI), 1.93-1.84 (m, 2 H), 1.33 (t J - 7.1 Hz, 3 H). Anal. (C18FI24N204) Calc: C, 65.04; H, 7.28; N, 8.43. Found: C, 64.70; H, 7.58; N, 8.56.
A solution of KOH (651 mg, 11.6 mmol) in water (10 mL) was added to a suspension of the ester 25 124 (811 mg, 2.44 mmol) in EtOH (20 mL). The mixture was stirred at reflux for 10 min then cooled to room temperature. The EtOH was evaporated and the aqueous residue was neutralised by the addition of aqueous HCl (2N, 5.80 mL, 11.6 mmol). An oil separated that solidified on standing. The solid was filtered off and dried, then suspended in dioxane (30 mL) and treated with HC1-saturated dioxane (10 mL). After stirring at room temperature for 3 h the solid was filtered off and 30 dried to give 5-(3-morpholinopropoxy)-lH-indole-2-carboxylic acid hydrochloride (125) as a white solid (824 mg, 99%): mp 246-249 °C; SH NMR [(CD3)2SO] 8 12.81 (br s, 1 H), 11.61 (s, 1 H), 10.69 (br s, 1 H), 7.34 (dj = 8.9 Hz, 1 H), 7.13 (dJ = 2.3 Hz, 1 H), 6.99 (ddj = 2.0, 0.6 Hz, 1 H), 6.91 (ddj = 8.9, 2.4 Hz, 1 H), 4.06 (tj = 6.0 Hz, 2 H), 4.01-3.92 (m, 2 H), 3.85-3.74 (m, 2 H), 3.53-3.43 (m, 2 H), 3.16-3.03 (m, 2 H), 2.24-2.14 (m, 2 H), (2 FI obscured by water peak observed after D20 48 Received at IPONZ on 14 December 2010 exchange). Anal. (C16H20N2O4'HCl'y2H2O) Calc: C, 54.94; H, 6.34; N, 8.01. Found: C, 55.00; H, 6.62; N, 7.64.
A mixture of indoline 108 (87 mg, 0.23 mmol), acid 125 (123 mg, 0.36 mmol), EDCI.HCl (173 mg, 5 0.92 mmol), and anhydrous TsOH (7.8 mg, 0.05 mmol) in DMA (2.5 mL) was stirred at room temperature for 3 h then cooled in an ice bath. Cold aqueous NaHC03 (5%) was added and the precipitated solid was filtered off, washed with water, and dried to give 2 as a yellow solid (153 mg, 100%): mp 130-134 °C; 'H NMR [(CD3)2SO] S 11.72 (dj = 1.7 Hz, 1 H), 9.29 (s, 1 H), 8.85 (dj = 1.6 Hz, 1 H), 8.44 (dJ = 8.9 FIz, 1 H), 8.04 (ddj = 8.9, 1.7 Hz, 1 H), 7.91 (tJ = 5.9 Hz, 1 H), 7.42 10 (dj — 8.9 Hz, 1 H), 7.22 (dJ = 1.8 Hz, 1 H), 7.16 (dj = 2.3 Hz, 1 H), 6.95 (ddj - 8.9, 2.4 Hz, 1 H), 5.01-4.94 (m, 1 H), 4.72 (ddj - 10.9, 2.4 Hz, 1 H), 4.69-4.63 (m, 1 H), 4.67 (tj = 5.6 Hz, 1 H), 4.21-4.10 (m, 2 H), 4.04 (t J = 6.4 FIz, 2 H), 3.63-3.57 (m, 4 H), 3.39 (q J = 6.0 Hz, 2 H), 2.88 (q J = 6.1 Hz, 2 H), 2.47 (t J = 7.2 Hz, 2 H), 2.43-2.36 (m, 4 FI), 1.96-1.88 (m, 2 H). Anal. (C31H34C1N50rS'H20) Calc: C, 53.95; H, 5.26; N, 10.15. Found: C, 53.97; FI, 5.21; N, 10.29.
Example 5. 5-Amino-l- (chioromethyl)-2V~ (2-hydroxyethyl)-3- [5- (3-morpholinopropoxy)-lH-indole-2-carbonyl]-l,2-dihydro-3//-benzo[e]indole-7-sulfonamide (11) (Scheme C). A solution of nitro compound 2 (52.6 mg, 0.078 mmol) in TFIF (20 mL) with Pt02 (53 mg) was hydrogenated at 50 psi for 2 h. The catalyst was filtered off through Celite, the filtrate was 20 evaporated, and the residue was triturated with EtOAc to give 11 as a pale green solid (38 mg, 76%): mp 220-225 °C (dec.); "'H NMR [(CD3)2SO] S 11.56 (d J = 1.5 Hz, 1 FI), 8.54 (d J = 1.6 Hz, 1 H), 7.94 (dj - 8.9 Hz, 1 H), 7.81 (s, 1 H), 7.74 (ddj = 8.9, 1.7 Hz, 1 FI), 7.49-7.44 (m, 1 H), 7.40 (dj - 8.9 Hz, 1 H), 7.16 (dj = 2.3 Hz, 1 H), 7.09 (dj= 1.5 Hz, 1 H), 6.92 (ddj - 8.9, 2.4 Hz, 1 H), 6.30 (s, 2 H), 4.76 (ddj = 10.7, 9.0 Hz, 1 H), 4.65 (tJ= 5.4 Hz, 1 H), 4.53 (ddj= 10.9, 1.7 Hz, 1 25 H), 4.21-4.15 (m, 1 H), 4.06-3.98 (m, 3 FI), 3.82 (dd J = 11.0, 7.7 Hz, 1 H), 3.63-3.56 (m, 4 H), 3.39 (qJ = 6.0 Hz, 2 H), 2.88-2.81 (m, 2 H), 2.46 (tj = 7.2 Hz, 2 H), 2.42-2.36 (m, 4 H), 1.95-1.87 (m, 2 H). Anal. (C3]H36C1N506S) Calc: C, 57.98; H, 5.65; N, 10.91. Found: C, 57.70; H, 5.70; N, 11.10.
Example 6. 2-{l-(ChIoromethyl)-3-[5-(3-morpholinopropoxy)-l//-indole-2-carbonyl]-5-nitro-30 l,2-dihydro-3il£benzo[e]indole-7-sulfonamido}ethyl dihydrogen phosphate trifluoroacetate (16) (Scheme C). A mixture of indoline 115 (176 mg, 0.30 mmol), acid 125 (156 mg, 0.45 mmol), EDCI.HCl (233 mg, 1.2 mmol), and anhydrous TsOH (10.5 mg, 0.06 mmol) in DMA (2 mL) was stirred at room temperature for 2 h then cooled in an ice bath. Cold aqueous NaHCOs (5%) was added and the precipitated solid was filtered off, washed with water, and dried. The resulting solid 35 was triturated with acetone to give di-/tr/-butyl 2-{l-(chloromethyl)-3-[5-(3-morpholinopropoxy)- 49 Received at IPONZ on 14 December 2010 lH-indole-2-carbonyl]-5-nitro-l ,2-dihydro-3i i-benzo [i?]indole-7-snlfonaniido } ethyl phosphate (126) as a pale yellow solid (175 mg, 66 %): mp 200-203 °C (dec.); 'H NMR [(CD3)2SO] 8 11.76 (d, J = 2.0 Hz, 1 H), 9.30 (s, 1 H), 8.87 (d J - 1.7 Hz, 1 H), 8.46 (d J - 8.9 Hz, 1 H), 8.20 (t J = 5.9 Hz, 1 H), 8.02 (ddj = 8.9, 1.7 Hz, 1 H), 7.42 (dJ = 8.9 Hz, 1 H), 7.23 (dj = 1.7 Hz, 1 H), 7.16 (dj = 2.3 5 Hz, 1 H), 6.95 (ddj = 8.9, 2.4 Hz, 1 H), 5.02-4.95 (m, 1 H), 4.74 (ddj = 10.9, 2.3 Hz, 1 H), 4.70-4.63 (m, 1 H), 4.20-4.11 (m, 2 H), 4.03 (tj = 6.3 Hz, 2 H), 3.84 (qJ = 6.3 Hz, 2 H), 3.62-3.56 (m, 4 H), 3.05 (qj = 5.8 Hz, 2 H), 2.46 (tj = 7.2 Hz, 2 H), 2.41-2.36 (m, 4 H), 1.35 (s, 18 H). Anal. (C^ClNsOuPS) Calc: C, 54.20; H, 5.95; N, 8.10. Found: C, 54.13; H, 5.91; N, 7.82.
TFA (0.14 mL, 1.9 mmol) was added to a suspension of the phosphate ester 126 (162 mg, 0.19 mmol) in CH,C12 (6 mL) and the resulting solution was stirred at room temperature for 16 h. The mixture was evaporated, the residue was resuspended in CH2C12 and evaporated once more, and the residue was triturated with EtOAc to give 16 as a pale yellow powder (165 mg, 100%): mp 165-168 °C; 'H NMR [(CD3)2SO] 8 11.82 (dj - 1.6 Hz, 1 H), ca. 9.7 (v br s, 1 H), 9.30 (s, 1 H), 8.87 (dJ = 15 1.6 Hz, 1 H), 8.44 (dj= 8.9 Hz, 1 H), 8.20 (tj= 5.9 Hz, 1 H), 8.02 (ddj= 8.9,1.7 Hz, 1 H), 7.44 (dj = 8.9 FIz, 1 H), 7.23 (dj = 1.7 Hz, 1 H), 7.20 (dj = 2.2 Hz, 1 H), 6.97 (ddj = 8.9, 2.4 Hz, 1 H), 5.02-4.93 (m, 1 H), 4.71 (ddj = 10.9, 2.2 Hz, 1 H), 4.68-4.62 (m, 1 H), 4.20-3.97 (m, 6 H), 3.81 (qj = 6.5 Hz, 2 H), 3.34-3.28 (m, 2 H), 3.20-3.08 (m, 2 H), 3.02 (qj = 5.9 Hz, 2 FI), 2.21-2.10 (m, 2 H), (4 H not observed, obscured by water peak). Anal. (C3,H,-CINsO /PS'CF ;CO2H'H2O) Calc: C, 20 44.83; H, 4.33; N, 7.92. Found: C, 44.96; H, 4.41; N, 8.20.
Example 7. (£)-l-(Chloromethyl)-Ar-(2-hydroxyetliyl)-3-{3-[4-(2- morphoIinoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3i/-benzo[e]indole-7-sulfonamide (3) (Scheme D). A mixture of 4-(2-chloroethyl)morpholine hydrochloride (6.42 g, 34.5 mmol), 25 NaOH (1.52 g, 38.1 mmol), water (24 mL) and toluene (26 mL) was stirred at 0 °C and saturated with solid NaCl. The toluene layer was separated and the alkaline aqueous solution was further extracted with toluene (4x15 mL). The combined toluene extracts were dried over KOFI and used for the following reaction.
To a solution of (E)-methyl 3-(4-hydroxyphenyl)acrylate (127) (5.43 g, 30.5 mmol) in anhydrous 30 ether (100 mL) stirred under nitrogen at 20 °C was added portionwise NaH (60% dispersion in oil, 1.28 g, 32.0 mmol). After the addition was complete the mixture was stirred for a further 2 h. Most of the ether was evaporated, petroleum ether (100 mL) was added, and the mixture was stirred for 5 min. The petroleum ether was decanted and the process was repeated once more. To the residue was added toluene (26 mL) followed by the toluene solution of 4-(2-chloroethyl)morpholine 3 5 prepared above. The heterogeneous mixture was stirred at reflux temperature for 92 h. The mixture 50 Received at IPONZ on 14 December 2010 was cooled and filtered through a short A1203 (neutral) column eluting with hot toluene. The combined eluates were washed successively with cold aqueous NaHC03 and then with water, then dried (Na2SO+) and evaporated to give (E)-methyl 3-[4-(2-morpholinoethoxy)phenyl]acrylate (128) as a colourless solid (6.66 g, 75%): mp 61-62 °C; 'H NMR (CDC13) 8 7.64 (dJ = 16.0 Hz, 1 H), 7.51-5 7.45 (m, 2 H), 6.93-6.88 (m, 2 H), 6.31 (dj = 16.0 Hz, 1 H), 4.14 (tJ = 5.7 Hz, 2 H), 3-79 (s, 3 H), 3.77-3.70 (m, 4 H), 2.81 (t J = 5.7 Hz, 2 H), 2.61-2.55 (m, 4 H). Anal. (C!6H21N04) Calc: C, 65.96; H, 7.27; N, 4.81. Found: C, 65.98; H, 7.33; N, 4.82.
A mixture of ester 128 (6.58 g, 22.6 mmol), MeOH (40 mL), water (20 mL) and KOH (3.16 g, 56.5 10 mmol) was stirred at 20 °C for 21 h. The mixture was evaporated under reduced pressure at 35 °C to remove the MeOH. The resulting alkaline mixture was washed with CH2C12 and the aqueous layer was filtered. The alkaline filtrate was acidified with conc. HCl at 0 °C. The resulting precipitate was filtered off, washed with 6N HCl and dried over silica gel under vacuum to give (E)-3-[4-(2-morpholinoethoxy)phenyl]acrylic acid hydrochloride (129) as a colourless solid (5.90 g, 83%): mp 15 252-254 °C; 'H NMR [(CD3)2SO] S 12.22 (br s, 1 H), 10.90 (br s, 1 H), 7.69 (dJ = 8.8 Hz, 2 H), 7.56 (d J = 16.0 Hz, 1 H), 7.05 (d J = 8.8 Hz, 2H), 6.41 (dj = 16.0 Hz, 1 H), 4.46 (br s, 2 H), 4.06-3.73 (m, 4 H), 3.67-3.42 (m, 4 H), 3.28-3.14 (m, 2 H). Anal. (C15H2UC1N04*0.3H20) Calc: C, 56.45; H, 6.51; N, 4.39. Found: C, 56.49; H, 6.46; N, 4.42.
A mixture of acid 129 (68.1 mg, 0.24 mmol), DMF (one drop) and thionyl chloride (2 mL) was stirred at reflux for 10 min. The mixture was cooled and evaporated under reduced pressure to dryness. The reaction flask was immersed in an ice-bath and solid indoline 112 (108 mg, 0.215 mmol) was added, followed by a mixture of DMA (2 mL) and DIPEA (0.041 mL, 0.237 mmol). The resulting mixture was stirred at 0 °C for 2 h. Dilute aqueous NaHCO, (5%, 7 mL) was added, 25 followed by water (7 mL), and the mixture was stirred at 0 °C for 10 min. The precipitated solid was filtered off, washed with cold water several times, then dried and triturated with EtOAc/petroleum ether (1:10) to give (B)-N-[2-(&nf-butyldimethylsilyloxy)ethyl]-l-(chloromethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3H-benzo[V]indole-7-sulfonamide (130) as a yellowish orange solid (128 mg, 79%): mp 203-205 °C; 'H NMR (CDC13) 8 9.42 (br s, 1 H), 8.99 (d, J 30 =1.6 Hz, 1 H), 8.03 (dd J = 8.9,1.6 Hz, 1 H), 7.95 (dj = 8.9 Hz, 1 H), 7.90 (d J = 15.2 Hz, 1 H), 7.58 (dj = 8.8 Hz, 2 H), 6.96 (dJ = 8.8 Hz, 2 FI), 6.74 (dJ = 15.2 Hz, 1 H), 4.96 (tj= 5.9, 1 H), 4.64 (ddj = 10.7, 2.4 Hz, 1 H), 4.56 (br tJ = 9.7 Hz, 1 H), 4.35-4.26 (m, 1 H), 4.18 (tj = 5.7, 2 H), 3.95 (br ddj =11.4, 3.3 Hz, 1 H), 3.80-3.72 (m, 4 H), 3.68 (tj = 5.1 Hz, 2 H), 3.63 (br ddj = 11.4, 9.5 FIz, 1 H), 3.22-3.13 (m, 2 H), 2.84 (t J = 5.7 Hz, 2 H), 2.65-2.55 (m, 4 H), 0.833 (s, 9 H), - Received at IPONZ on 14 December 2010 0.008 (s, 6 H, obscured by TMS). Anal. (C36H47ClN408SSi) Calc: C, 56.94; H, 6.24; N, 7.38. Found: C, 57.09; H, 6.14; N, 7.27.
To a mixture of silyl ether 130 (142 mg, 0.187 mmol), dioxane (6 mL) and MeOFI (3 mL) stirred at 5 20 °C was added HCl (1.25M in MeOH, 1.0 mL). The mixture was stirred at this temperature for a further 30 min. EtOAc was added and the mixture was stirred for another 30 min. The resulting solid was filtered off, washed with EtOAc several times, and dried to give 3 as a yellowish orange solid (127 mg, 99%); mp 280-282 °C (dec.); ]H NMR [(CD3)2SO] S 10.75 (br s, 1 H), 9.35 (s, 1 H), 8.83 (dJ = 1.6 Hz, 1 H), 8.38 (dj = 8.9 Hz, 1 H), 8.02 (ddj = 8.9, 1.6 Hz, 1 H), 7.92 (t,J = 6.0 10 Hz, 1 H), 7.84 (dj= 8.7 Hz, 2 H), 7.75 (dj= 15.3 Hz, 1 H), 7.14 (dj= 15.3 Hz, 1 H), 7.09 (dj = 8.7 Hz, 2 H), 4.72-4.60 (m, 3 H), 4.49 (br s, 2 H), 4.10 (br dj = 4.1 Hz, 2 H), 3.99 (br dJ = 13.8, 2 H), 3.80 (br t J = 11.7 Hz, 2 H), 3.63-3.49 (m, 4 H), 3.39 (t J = 6.2 Hz, 2 H, partially obscured by water peak), 3.30-3.16 (m, 2 H, partially obscured by water peak), 2.87 (q J — 6.0 Hz, 2 H). HRMS (FAB) calc for C30H3435C1N4OsS (MH+) m/% 645.1785, found 645.1787. Calc for C30FI3437ClN4O8S 15 (MH+) jw/? 647.1756, found 647.1770. Anal. (C30H34Cl2N4O8S-H2O-Y2EtOAc) Calc: C, 51.68; H, 5.42; N, 7.53. Found: C, 51.41; H, 5.31; N, 7.50.
Example 8. (£)-5-Amino-l-(chlorometliyl)-At-(2-hydroxyethyl)-3-{3-[4-(2-morph.olinoethoxy)phenyl] acryloyl}-l,2-dihydro-3.fif-benzo[e]indole-7-sulfonamide (12) 20 (Scheme D). Saturated aqueous ammonium chloride (4 ml.,) and then Zn powder (450 mg) were added to a stirred solution of nitro compound 130 (107 mg, 0.141 mmol) in acetone (8 mL) and water (4 mL). The mixture was stirred at room temperature for 1 h, then filtered through Celite and the Celite pad was rinsed with acetone. The combined filtrates were concentrated to remove acetone and the aqueous residue was extracted with dichloromethane (x 2). The organic extracts were 25 washed with water and then brine, and then dried (Na2S04) and evaporated. The residue was dissolved in dioxane (1.5 mL) and MeOH (0.75 mL), and HCl (4M in MeOH, 1 mL) was added. The resulting mixture was stirred at room temperature for 30 min, concentrated under reduce pressure, and diluted with CH2C12 (2 mL). The mixture was cooled at 0 °C and ice-cold aqueous NaHC03 (10%, 10 mL) was added, followed by water (10 mL). The mixture was stirred for 30 min then 30 extracted with CH2C12 (x 2). The organic extracts were washed with water and then brine, and then dried (Na2S04) and evaporated. The residue was dissolved in the minimum quantity of MeOH, cooled to 0 °C, and diluted with EtzO. After stirring at 0 °C for 30 min the resulting precipitate was filtered off and washed with water and then Et2C) to provide 12 as a yellow powder (38 mg, 44%): mp 225-230 °C (dec.); *H NMR [(CD3)2SO] 8 8.51 (dj = 1.5 Hz, 1 H), 7.89-7.84 (m, 2 H), 7.76-7.68 35 (m, 3 H), 7.63 (dj = 15.3 Hz, 1 H), 7.41 (br s, 1 H), 7.06 (dj = 15.6 Hz, 1 FI), 7.01 (dj = 8.8 Hz, Received at IPONZ on 14 December 2010 2 H), 6.25 (s, 2 H), 4.63 (tJ = 5.6 Hz, 1 H), 4.51-4.38 (m, 2 H), 4.16-4.02 <m, 3 H), 3.97-3.92 (ddj = 10.9, 2.9 Hz, 1 H), 3.81-3.76 (ddj = 11, 8 Hz, 1 H), 3.61-3.56 (m, 4 H), 3.38 (qJ = 12, 6.3 Hz, 2 H), 2.83 (m, 2 H), 2.71 (t J — 5.8 Hz, 2 H), 2.53-2.46 (4 H obscured by DMSO peak). Anal. (C30H35C1N406S-0.2H20) Calc: C, 58.24; H, 5.77; N, 9.06. Found: C, 58.31; H, 5.63; N, 8.74.
Example 9. (.£)-2-(l-(Chloromethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl] acryloyl}-5-nitro-l,2-dihydro-3J££benzo[e]mdole-7-sulfonamido) ethyl dihydrogen phosphate trifluoroacetate (17) (Scheme D). To a stirred mixture of indoline 115 (290 mg, 0.50 mmol) and acid 129 (198 mg, 0.60 mmol) in DMA (10 mL) was added EDCI.HCl (385 mg, 2.00 mmol) and anhydrous TsOH 10 (17.5 mg, 0.10 mmol). The mixture was stirred at 20 °C for 4 h and another batch of acid 129 (198 mg, 0.60 mmol), EDCI.HCl (385 mg, 2.00 mmol), and anhydrous TsOH (17.5 mg, 0.10 mmol) were added. After a total reaction time of 24 h the reaction flask was cooled in an ice-bath and cold aqueous NaHC03 (5%, 20 mL) was added, followed by water (20 mL). After stirring at 0 °C for 10 min the precipitated solid was filtered off, washed with cold water several times, and dried to give 15 (JE^-di-fert'-butyl 2-(l-(chloromethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloy}-5-mtro-l,2-dihydro-3H-benzo[tf]indole-7-sulfonamido)ethyl phosphate (131) as a yellowish orange solid (395 mg, 94%): mp 245-248 °C (dec.); 'H NMR (CDC13) 5 9.41 (br s, 1 H), 8.97 (dj = 1.6 Hz, 1 H), 8.50 (dd, J = 8.9, 1.6 Hz, 1 H), 7.96 (dj = 8.9 Hz, 1 H), 7.91 (dj= 15.2 Hz, 1 H), 7.58 (dj= 8.7 Hz, 2 FI), 6.96 (d J = 8.7 Hz, 2 H), 6.74 (d J = 15.2 Hz, 1 H), 5.95 (t J = 5.7 Hz, 1 H), 4.63 (brddj = 20 10.8, 2.5 FIz, 1 FI), 4.56 (br t J = 9.8 Hz, 1 H), 4.34-4.26 (m, 1 H), 4.18 (t J = 5.6, 2 H), 4.12-4.04 (m, 2 H), 3.96 (br ddj = 11.5, 3.2 Hz, 1 H), 3.75 (br t J = 4.6 Hz, 4 H), 3.62 (br dd J = 11.4, 9.9 Hz, 1 H), 3.33 (br q J - 5.4 Hz, 2 H), 2.85 (br t J = 5.6 Hz, 2 H), 2.61 (unresolved t, 4 H), 1.46 and 1.45 (2 s, 18 H). Anal. (C^H^a^O^S'yzHp) Calc: C, 53.93; H, 6.07; N, 6.62. Found: C, 53.83; H, 5.99; N, 6.53.
Phosphate ester 131 (366 mg, 0.437 mmol) was stirred with TFA (5 mL) in CH2C12 (50 mL) at 20 °C for 3 h. The reaction solution was filtered through a cotton wool plug and the plug was washed with methanol. The filtrate was concentrated under reduced pressure until a precipitate began to form. EtOAc (150 mL) was added and the mixture was stirred at 0 °C for 45 min. The solid was filtered 30 off, washed with EtOAc several times, and dried in vacuum over silica gel to give 17 as a yellow solid (292 mg, 80%): mp 212 °C (dec.); ]H NMR (C5DSN) 8 ca. 10.7 (v br s, 1 H), 9.81 (br s, 1 H), 9.46 (dj = 1.6 Hz, 1 H), 8.39 (ddj = 8.9,1.6 Hz, 1 H), 8.22 (dj = 15.2 Hz, 1 H), 8.14 (dj = 8.9 Hz, 1 H), 7.74 (dj = 8.8 Hz, 2 H), 7.16 (dJ = 15.2 Hz, 2 H), 7.10 (dJ = 8.8 FIz, 2 H), 4.80 (br dd, J = 10.8,2.6 Hz, 1 H), 4.65 (br t J = 10.0 Hz, 1 H), 4.59-4.51 (m, 2 H), 4.50-4.42 (m, 1 H), 4.21-4.13 (m, 3 H), 4.01 (br ddj =11.3, 8.3 Hz, 1 H), 3.73 (br tJ = 4.6 Hz, 6 H), 2.76 (tJ = 5.7 Hz, 2 H), 53 Received at IPONZ on 14 December 2010 2.53 (t ,J = 4.6 Hz, 4 H). Anal. (C^ClNp^PS'VaCF^O^) Calc: C, 47.61; H, 4.45; N, 7.16. Found: C, 47.69; H, 4.65; N, 6.97.
Example 10. l-(ChIoromethyl)-AL(2-hydfoxyethyl)-3-{4-[2-(4-methylpipera2in-l-5 yl)ethoxy]benzoyl}-5-nitro-l,2-dihydro-3//-benzo[e]indole-7-sulfonamide hydrochloride (4) (Scheme E). To a solution of (E)-methyl 3-(4-hydroxyphenyl)acrylate (127) (5.4 g, 30.5 mmol) in anhydrous Et20 (100 mL) was added NaH (0.77 g, 32.0 mmol) and the mixture was stirred under nitrogen at 20 °C for 6 h. The Et,G was evaporated and DMA (20 mL) was added, followed by 1,2-dibromoethane (13.2 mL, 153 mmol). The reaction mixture was stirred at 130-135 °C (bath 10 temperature) for 21 h. The mixture was cooled to 20 °C and more NaH (0.77 g, 32.0 mmol) and 1,2-dibrotnoethane (13.2 mL, 153 mmol) were added. The mixture stirred again at 150-160 °C (bath temperature) for 5 h. The mixture was cooled a little and toluene (100 mL) was added. The mixture was filtered and the filter pad was washed with hot toluene several times. The combined filtrates were washed successively with cold water, cold aqueous NaPICO,, and again with water. The 15 organic layer was dried (Na2S04) and evaporated to give an oil which was dissolved in DMF (30 mL) and stirred with 1-methylpiperazine (14.7 mL, 133 mmol) at 20 °C for 3 h. The mixture was diluted with EtOAc (ca. 200 mL) and then stirred with aqueous Na2CO, (2%) at 0 °C for 5-10 min. The organic layer was separated, washed with water (x 2), and then dried (Na2S04) and evaporated. The resulting oil was dissolved in EtOAc and the solution was treated with HCl (4M in p-dioxane, 17 20 mL, 68 mmol). The mixture was stirred and then left to stand at 5 °C over-night. The solid was filtered off, washed with EtOAc several times, and dried to give (E)-methyl 3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}acrylate hydrochloride (132) as beige solid (5.47g, 48%): mp 235-237 °C (dec.); ]H NMR [(CD3)2SO] 8 11.78 (br s, 2 H), 7.71 (d J = 8.8 Hz, 2 H), 7.64 (d J = 16.0 Hz, 1 H), 7.05 (d ,J — 8.8 Hz, 2 FI), 6.52 (d,J — 16.0 Hz, 1 H), 4.44 (poorly resolved t resolved 25 after D20 exchange, J = 4.9 Hz, 2 H), 3.71 (s, 3 FI), 3.52-3.31 (m, partially obscured by water peak, revealed after D20 exchange, 10 H), 2.81 (s, 3 H). Anal. (C17H26C12N203 VzHjO) Calc: C, 52.86; H, 7.05; N, 7.25. Found: C, 52.79; FI, 6.98; N, 6.94.
Ester 132 (5.18 g, 13.7 mmol) was treated with KOH (3.5 g, 62 mmol) in a mixture of MeOH (30 mL) and 30 water (15 mL) at 20 °C for 29 h. The mixture was evaporated under reduced pressure at 35 °C to remove the MeOH. The alkaline remainder was washed with CH2C12 and filtered. The filtrate was acidified with conc. FIC1 at 0 °C and left to stand at 5 °C overnight. The resulting precipitate was filtered off, washed with cold 6N HCl (3x1 mL), and dried over silica gel in vacuum overnight to give (B)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]phenyl}acrylic acid hydrochloride (133) as a beige 35 solid (2.97 g, 60%): mp 264-267 °C (dec.); 1H NMR f(CD3)2SO] 8 11.89 (br s, 3 H), 7.67 (dJ = 8.8 54 Received at IPONZ on 14 December 2010 Hz, 2 H), 7.56 (dj= 16.0 Hz, 1 H), 7.05 (dJ = 8.8 Hz, 2 H), 6.41 (dJ = 16.0 Hz, 1 H), 4.46 (unresolved t resolved after D20 exchange, / = 4.8 Hz, 2 H), 3.71 (s, 3 H), 3.57-3.30 (m, partially obscured by water peak, revealed after D20 exchange, 10 H), 2.82 (s, 3 H). Anal. (C16FI24C12N203V2H20) Calc: C, 51.62; H, 6.77; N, 7.53. Found: C, 51.77; H, 6.63; N, 7.17.
To a stirred mixture of indoline 112 (90 mg, 0.18 mmol) and acid 133 (78 mg, 0.22 mmol) in DMA (10 mL) were added EDCI.HCl (138 mg, 0.72 mmol) and anhydrous TsOFI (6.3 mg, 0.04 mmol). The mixture was stirred at 20 °C for 3 h then another batch of EDCI.HCl (138 mg, 0.72 mmol) and anhydrous TsOH (6.3 mg, 0.04 mmol) were added and the mixture was stirred for a 10 further 16 h. The reaction flask was immersed in an ice-bath and cold aqueous NaHCOj (5%, 10 mL) was added, followed by water (30 mL). The mixture was stirred at 0 °C for ca. 10 min. The solid was filtered off, washed with cold water several times, and dried to give N-\2-(/cr/-butyldimethylsilyloxy) ethyl] -1 -(chloromethyl) -3- {4-[2- (4-methylpip erazin-1 -yl) ethoxvjbenzoyl} -5-nitro-l,2-dihydro-3H-benzo[£]indole-7-sulfonatrdde (134) as a yellowish orange solid (118 mg, 85%): 15 mp 202 °C (dec.); 'H NMR (CDC1,) 8 9.42 (br s, 1 H), 8.98 (poorly resolved dj = 1.6 Hz, 1 H), . 8.03 (ddj = 8.8, 1.6 Hz, 1 H), 7.95 (dj= 8.8 Hz, 1 FI), 7.91 (dj = 15.2 Hz, 1 H), 7.57 (dj = 8.7 Hz, 2 H), 6.95 (dj - 8.7 Hz, 2 H), 6.73 (dj = 15.2 Hz, 1 H), 4.99 (tj= 5.8 Hz, 1 H), 4.63 (ddJ-10.8, 2.5 Hz, 1 H), 4.45 (br t J = 9.7 FIz, 1 H), 4.34-4.26 (m, 1 H), 4.17 (t J = 5.8 Hz, 2 H), 3.94 (dd, /= 11.4, 3.3 Hz, 1 H), 3.69 (tJ = 5.0 FIz, 2 H), 3.62 (ddj =11.4, 9.5 Hz, 1 FI), 3.22-3.13 (m, 2 H), 20 2.85 (tj = 5.8 Hz, 2 FI), 2.73-2.42 (m, 8 H), 2.30 (s, 3 H), 0.84 (s, 9 FI), -0.008 (s, 6 H, obscured by TMS). Anal. (C37H5ilClN507SSi) Calc: C, 57.53; H, 6.52; N, 9.07. Found: C, 57.51; H, 6.52; N, 8.86.
HCl (1.25M in MeOH, 1 mL) was added to a solution of silyl ether 134 (98 mg, 0.13 mmol) in CH2C12 (15 mL) and the mixture was stirred at 20 °C for 2 h 20 min. EtOAc (80 mL) was added and 25 the mixture was stirred at 0 °C for 2 h. The resulting solid was filtered off, washed with EtOAc, and dried to give 4 as a yellowish orange solid (93 mg, 100%): mp 225 °C (dec.); NMR [(CD3)2SO] 8 11.10 (v br s, exchangeable with D20,1 H), 9.35 (br s, 1 H), 8.83 (poorly resolved d J = 1.6 Hz, 1 H), 8.39 (d J = 8.9 Hz, 1 H), 8.02 (dd J = 8.9, 1.6 Hz, 1 H), 7.92 (t J = 5.9 Hz, exchangeable with D20, 1 H), 7.83 (dj = 8.7 Hz, 2 H), 7.74 (dj = 15.3 Hz, 1 H), 7.13 (dj= 15.3 Hz, 1 H), 7.08 (dj 30 = 8.7 Hz, 2 H), 4.72-4.59 (m, 3 H), 4.39 (br s, 2 H), 4.08 (poorly resolved d J = 4.0 Hz, 2 H), 3.38 (tj — 6.2 Hz, 2 H), 3.35-2.91 (m, revealed after D20 exchange, 10 H), 2.93-2.75 (m, 5 H). Anal. (C31H38C1,N507S0.9H20) Calc: C, 49.83; H, 5.37; N, 9.37. Found: C, 50.14; H, 5.32; N, 9.03.
Example 11. 2-[l-(Chloromethyl)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-3 5 l,2-dihydro-3/£benzo [ e] indole-7-sulfonamido] ethyl dihydrogen phosphate trifluoroacetate 55 Received at IPONZ on 14 December 2010 (18) (Scheme E). To a stirred mixture of indoline 115 (1.16 g, 2.00 mmol) and acid 133 (0.87 g, 2.40 mmol) in DMA (45 mL) was added EDCI.HCl (1.54 g, 8.00 mmol) and anhydrous TsOH (70.4 mg, 0.40 mmol). The mixture was stirred at 20 °C for 4 h and then another batch of acid 133 (0.87 g, 2.40 mmol), EDCI.HCl (1.54 g, 8.00 mmol) and anhydrous TsOH (70.4 mg, 0.40 mmol) were added 5 and the mixture was stirred for a further 18 h. The reaction flask was immersed in an ice-bath and cold aqueous NaHC03 (5%, 90 mL) was added, followed by water (90 mL). The mixture was stirred at 0°C for 10 min. The precipitated solid was filtered off, washed with cold water several times, and dried to give di-Zfi^-butyl 2-[l-(chloromethyl)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-l,2-dihydro-3J-T-benzo[^]indole-7-sulfonamido]ethyl phosphate (135) as a yellowish orange 10 solid (1.58 g, 93%): mp 118-120 °C (dec.); NMR (CDC13) 8 9.41 (br s, 1 EI), 8.98 (poorly resolved dj = 1.3 Hz, 1 H), 8.04 (dd,/ = 8.8, 1.4 Hz, 1 H), 7.95 (d,/= 8.8 Hz, 1 H), 7.90 (dJ = 15.2 Hz, 1 H), 7.57 (d, / = 8.6 Hz, 2 H), 6.94 (dj = 8.6 Hz, 2 H), 6.73 (dJ = 15.2 Hz, 1 H), 5.96 (br s, 1 H), 4.63 (ddj = 10.7, 2.3 Hz, 1 H), 4.55 (br tJ = 9.7 Hz, 1 H), 4.32-4.23 (m, 1 H), 4.17 (tj = 5.8 Hz, 2 H), 4.12-4.02 (m, 2 H), 3.95 (dd, / = 11.4, 3.2 Hz, 1 H), 3.62 (ddj - 11.3, 9.5 Hz, 1 H), 3.32 (br s, 2 15 H), 2.85 (t J = 5.8 Hz, 2 H), 2.66 (br s, 4 H), 2.52 (br s, 4 H), 2.32 (s, 3 FI), 1.46 and 1.45 (2 s, 18 H). Anal. (C39H53ClN5O10S-V2H2O) Calc: C, 54.51; H, 6.33; N, 8.15. Found: C, 54.52; H, 6.34; N, 7.91.
Phosphate ester 135 (1.57 g, 1.85 mmol) was dissolved in CH2C12 (100 mL) and stirred with TFA (10 20 mL) at 20 °C for 3 h. The mixture was filtered and the filter pad was washed with 10% TFA in CH2C12. The combined filtrates were concentrated at 25 °C (bath temperature) under reduced pressure at to half of the original volume. EtOAC (ca. 100 mL) was added, the mixture was stirred and then left to stand at 0 °C for 15 h. The precipitated solid was filtered off, washed with EtOAc several times, and dried to give 18 as a yellowish orange solid (1.37 g, 87%): mp 230 °C (dec.); 'FI 25 NMR [(CD3)2SO] S 9.35 (br s, 1 H), 9.35 (br s, 1 H), 8.85 (poorly resolved d J - 1.6 Hz, 1 H), 8.38 (dj = 8.9 Hz, 1 H), 8.13 (t J = 5.9 FIz, exchangeable with D20, 1 H), 8.02 (ddJ = 8.9, 1.6 Hz, 1 H), 7.82 (dJ = 8.8 FIz, 2 FI), 7.74 (dj = 15.2 Hz, 1 H), 7.11 (dj - 15.2 Hz, 1 H), 7.04 (dj = 8.8 Hz, 2 H), 4.12-4.58 (m, 3 H), 4.21 (t J = 5.2 Hz, 2 H), 4.09 (poorly resolved d J = 4.9 Hz, 2 H), 3.82 (q J = 6.2 Hz, 2 H), 3.51-2.85 (m, partially obscured by water peak, revealed after D20 30 exchange, 12 H), 2.77 (s, 3 H), 2 Hs not observed. Anal. (C31H37ClNsO10P'1.5CF3COOH'EtOAc) Calc: C, 45.77; H, 4.70; N, 7.02. Found: C, 45.72; H, 4.67; N, 7.09.
Example 12. l-(Chloromethyl)-A£(2-hydroxyethyl)-3- [5-(2-niorpholinocthoxy)-l//-indolc-2-carbonyl]-5-nitro-l,2-dihydro-3//-bcnzo[c]indole-7-carboxamide (5) (Scheme F). A solution 35 of compound 110 (193 mg, 0.623 mmol) in THF (15 mL) was hydrogenated over Pd/C (5%) at 40 56 Received at IPONZ on 14 December 2010 psi for 2 h 30 min. The mixture was filtered through Celite and the Celite was washed with THF (5 x 10 mL). The combined THF filtrate (containing amine 111) was added to an ice-cold mixture of 1 -(chloromethyI)-5-nitro-3-(2,2,2-trifluoroacetyl)-l,2-dihydro-3H-benzo[£]indole-7-carboxylic acid (136) (209 mg, 0.519 mmol) and DIPEA (0.27 mL, 1.56 mmol) in THF (3 mL), followed by the 5 addition of pyBOP (benzotnazoI-l-yloxy)tripyrtoHdinophosphoniuin hexafluorophosphate) (352 mg, 0.675 mmol). The mixture was stirred at 0 °C for a further 1 h. Cesium carbonate (339 mg, 1.04 mmol) and MeOH (10 mL) were added and the mixture was stirred at 20 °C for 2 h, then allowed to stand at 5 °C overnight. The mixture was concentrated under reduced pressure at 25 °C to remove most of the THF. Ice-water was added and the mixture was stirred at 0 °C for 5 h 30 min. The 10 resulting solid was filtered off, washed with cold water several times, and dried in vacuum over silica-gel to give a crude product This was purified by column chromatography (eluting with CH2Cl2:EtOAc 5:1) to give pure JV-(2-(^-butyldimethylsilyloxy)ethyl)-l-(chloromethyl)-5-nitro-l,2-dihydro-3ii-benzo[t'jmdole-7-carboxamicIe (137) as a red solid (212 mg, 88%): mp 156-158 "C; 'EI NMR (CDC1,) 8 8.75 (poorly resolved dj = 1.4 FIz, 1 H), 8.02 (ddj = 8.8, 1.7 Hz, 1 H), 7.77 (d J 15 =8.8 Hz, 1 FI), 7.71 (s, 1 H), 6.67 (br s, 1 H), 4.26 (s, 1 H), 4.14-4.07 (m, 1 H), 4.01-3.91 (m, 2 H), 3.87-3.77 (m, 3 H), 3.67-3.55 (m, 3 H), 0.92 (s, 9 H), 0.11 (s, 6 H). Anal. (C^ClN^Si) Calc: C, 56.94; H, 6.52; N, 9.06. Found: C, 57.07; H, 6.67; N, 9.20.
To a stirred mixture of 137 (126 mg, 0.27 mmol) and acid 122 (107 mg, 0.33 mmol) in DMA (5 mL) 20 was added EDCI.PIC1 (210 mg, 1.09 mmol), and anhydrous TsOH (9.5 mg, 0.054 mmol). The mixture was stirred at room temperature for 2.5 h then cooled in an ice bath. Cold aqueous NaHCO, (5%, 10 mL) was added, followed by cold water (10 mL). The precipitated solid was filtered off, washed with water, and dried to give iV-[2-(/e^-butyldimethylsilyloxy)ethyl]-l-(chloromethyl)-3 - [5-(2-morpholinoethoxy)-1 H-indole-2-carbonyl]-5-nitro-1 ;2-dihydro-3H-25 benzo[<?]indole-7-carboxamide (138) as a yellow solid (193 mg, 97%): mp 124 °C; 'H NMR (CDCL) 8 9.38 (br s, 1 H), 9.28 (s, 1 FI), 8.79 (poorly resolved dj= 1.3 Hz, 1 FI), 8.15 (ddj = 8.8, 1.6 Hz, 1 H), 7.94 (dj = 8.8 Hz, 1 H), 7.38 (dj = 8.9 Hz, 1 H), 7.14 (poorly resolved dJ = 2.3 Hz, 1 H), 7.09-7.04 (m, 2 H), 6.79 (tJ - 5.2 Hz, 1 H), 4.92 (ddj =11.3, 2.1 Hz, 1 H), 4.81 (br tJ = 9.8 Hz, 1 H), 4.39-4.29 (m, 1 H), 4.18 (tJ = 5.7 Hz, 1 H), 3.99 (ddj= 11.4, 3.2 Hz, 1 H), 3.86 (tj= 5.2 Hz, 30 2 H), 3.82-3.74 (m, 4 H), 3.71-3.59 (m, 3 H), 2.86 (t J = 5.7 Hz, 2 H), 2.67-2.58 (m, 4 H), 0.94 (s, 9 H), 0.12 (s, 6 H). Anal. (C37H46ClN507Si0.3H20) Calc: C, 59.91; H, 6.33; N, 9.44. Found: C, 59.76; H, 6.41; N, 9.58.
To a filtered solution of 138 (83 mg, 0.113 mmol) in CHjC^ (10 mL) was added HCl (1.25M in 35 MeOH, 1.0 mL) and the mixture was stirred at 20 °C for 2 h. The mixture was diluted with EtOAc Received at IPONZ on 14 December 2010 (60 mL) and the precipitated solid was filtered off, washed with EtOAc several times, and dried to give 5 as a yellowish orange hydrochloride salt (69 mg, 93%): mp 214-217 °C; 1H NMR [(CD3)2SO] 8 11.80 (s, exchangeable with D20, 1 EI), 10.70 (s, exchangeable with D20, 1 H), 9.16 (s, 1 H), 8.82 (poorly resolved dJ =1.3 Hz, 1 H), 8.78 (t J = 5.6 Hz, exchangeable with DaO, 1 H), 8.30 (dJ = 5 8.8 Hz, 1 H), 8.16 (ddj = 8.8,1.5 Hz, 1 H), 7.46 (dJ = 8.9 Hz, 1 H), 7.28 (br s, 1 H), 7.22 (poorly resolved dJ = 1.6 Hz, 1 H), 7.04 (poorly resolved ddj = 8.9, 2.0 Hz, 1 H), 4.96 (br tJ = 10.1 Hz, 1 H), 4.70 (poorly resolved ddj = 10.9, 2.3 Hz, 1 H), 4.68-4.59 (m, 1 H), 4,43 (s, 2 H), 4.19-4.07 (m, 2 H), 4.05-3.71 (m, 4 H), 3.69-3.48 (m, 5 H, 1 H exchangeable with D20), 3.40 (q, collapses to t after D20 exchange J = 5.9 Hz, 2 H), 3.36 (m, partially obscured by water peak, revealed after D20 exchange, 4 H). Anal. (C31HffiClNs0v-HCl-y2H20) Calc: C, 55.78; H, 5.13; N, 10.49. Found: C, 55.65; H, 5.12; N, 10.43.
Example 13. 5-Amino-l-(chloromethyl)-A£(2-hydroxyethyl)-3-[5-(2-m.orpholinoethoxy)-lii-indole-2-carbonyI]-l,2-dihydro-3i/-benzo[e]indole-7-carboxamide (13) (Scheme F). A solution of nitro compound 138 (39.4 mg, 0.054 mmol) in THF (20 mL) with PtOz (60 mg) was hydrogenated at 40 psi for 15 min. The catalyst was filtered off through Celite and the filtrate was stirred with HCl (1.25M in MeOH, 0.5 mL) at 20 °C for 1 h. The mixture was evaporated and the residue was triturated with EtOAc to give 13 as a pale green hydrochloride salt (30 mg, 89%): mp >300 °C; 'H NMR (C5D5N) S 12.98 (s, 1 H), ca. 11.0 (v br s, 1 H), 9.59 (poorly resolved d J = 1.3 20 Hz, 1 H), 9.35 (t J = 5.4 Hz, 1 FI), 8.55 (br s, 1 H), 8.43 (poorly resolved dd J — 8.7, 1.5 Hz, 1 H), 7.86 (dj = 8.7 Hz, 1 FI), 7.68 (dj = 8.8 Hz, 1 H), 7.41 (dj = 2.3 FIz, 1 H), 7.34-7.24 (m, 2 H), 4.93 (poorly resolved ddj = 10.8, 1.7 Hz, 1 H), 4.73 (br tJ = 8.8 Hz, 1 H), 4.36 (tJ = 5.7 Hz, 2 H), 4.23-4.12 (m, 3 H), 4.10-3.96 (m, 3 H), 3.87-3.75 (m, 4 H), 3.65 (ddj = 10.9, 9.5 Hz, 1 H), 2.93 (t J = 5.7 Hz, 2 H), 2.73-2.63 (m, 4 H), 3 H not observed (obscured by water peak). HRMS (FAB) 25 calc for C31H3535C1N505 (MH+) tn/% 592.2327, found 592.2318. For C31H3537C1N505 (MFI+) m/^ 594.2297, found 594.2310.
Example 14. 2-{l-(Chloromethyl)-3- [5-(2-morphoIinoethoxy)-l//-mdolc-2-carbonyl]-5-nitro- I,2-dihydro-3Jy;-benzo[eJindole-7-carboxamido}ethyl dihydrogen phosphate hydrochloride 30 (19) (Scheme F). A solution of amine 114 (218 mg, 0.86 mmol) in THF (3 mL) was added to an ice-cold mixture of acid 136 (290 mg, 0.72 mmol), DIPEA (0.38 mL, 2.16 mmol) and THF (7 mL), followed by the addition of pyBOP (486 mg, 0.93 mmol). The resulting mixture was stirred at 0 °C for a further 50 min. Cesium carbonate (470 mg, 1.44 mmol) and MeOH (4 mL) were added and the mixture was stirred at 0 °C for 30 min, then allowed to stand at 5 °C overnight. The mixture was 35 partitioned between CH,C12 and cold water. The organic layer was separated and the aqueous Received at IPONZ on 14 December 2010 portion was further exacted with CFI,C12 twice. The combined CH2C12 layers were dried (Na2S04) and evaporated under reduced pressure at 25 °C (bath temperature) to give a crude product. This was purified by column chromatography (elutingwith CH2Cl2:EtOAc 2:1) to give pure di-fe/2-butyl 2-[l-(chloromethyl)-5-nitro-l,2-dihydro-3jFi-benzo|V]indole-7-carboxamido]ethyl phosphate (139) as 5 a red solid (194 mg, 50%): *H NMR [(CD3)2SO] 8 8.76 (tJ — 5.5 Hz, 1 H), 8.58 (poorly resolved d, J = 0.78 Hz, 1 H), 7.93 (ddj= 8.9,1.5 Hz, 1 H), 7.91 (dj = 8.7 Hz, 1 H), 7.65 (s, 1 H), 4.26-4.18 (m, 1 H), 4.00 (qJ = 6.1 Hz, 2 H), 3.91 (dd J = 11.0, 3.8 Hz, 1 H), 3.85 (ddj = 9.9, 2.2 Hz, 1 PI), 3.77 (dd, J = 11.0, 8.6 Hz, 1 H), 3.72 (ddj = 10.3, 2.9 Hz, 1 H), 3.52 (qJ = 5.6 Hz, 2 H), 1.39 (s, 18 H). HRMS (FAB) calc for C24FI3335C1N307P (MH+) m/% 541.1745, found 541.1743. Calc for 10 C24H3337CIN307P (MH+) m!^ 543.1715, found 543.1731.
To a stirred mixture of indoline 139 (143 mg, 0,26 mmol) and acid 122 (104 mg, 0.32 mmol) in DMA (5 mL) was added EDCI.HCl (204 mg, 1.06 mmol), and anhydrous TsOFI (9.3 mg, 0.053 mmol). The mixture was stirred at room temperature for 3 h then cooled in an ice bath. Cold 15 aqueous NaHC03 (5%, 10 mL) was added, followed by cold water (10 mL), The precipitated solid was filtered off, washed with water, and dried to give di-^-butyl 2-[l-(chloromethyl)-3-[5-(2-morpholinoethoxy) -1 H-indole-2-carbonyl] - 5-nitro-1,2-dihychro-3i-f-benzo [e] indole-7 -carboxamido]ethyl phosphate (140) as a yellowish orange solid (213 mg, 99%): mp 130-132 °C; 'H NMR (CDC13) S 9.34 (br s, 1 H), 9.24 (s, 1 H), 8.93 (poorly resolved dj = 1.3 Hz, 1 H), 8.20 (ddj 20 = 8.8, 1.6 Hz, 1 H), 8.06 (t J = 4.9 Hz, 1 H), 7.92 (dJ = 8.8 Hz, 1 H), 7.38 (dj = 8.9 Hz, 1 H), 7.14 (poorly resolved dJ = 2.3 Hz, 1 H), 7.10-6.99 (m, 2 H), 4.91 (ddj = 10.8, 2.1 Hz, 1 H), 4.80 (br tj = 9.7 Hz, 1 H), 4.38-4.13 (m, 5 H), 3.98 (ddj = 11.5, 3.3 Hz, 1 H), 3.86-3.72 (m, 6 H), 3.61 (ddj =11.4, 9.5 Hz, 1 H), 2.87 (tj = 5.6 Hz, 2 H), 2.70-2.58 (m, 4 H), 1.503 and 1.502 (2 s, 18 H). Anal. (C39H49C1N5010P-0.3H20) Calc: C, 57.15; H, 6.10; N, 8.54. Found: C, 57.02; FI, 6.06; N, 8.44.
To a stirred solution of 140 (197 mg, 0.242 mmol) in CPI2C12 (10 mL) was added HCl (saturated solution in CH2C12, 1.0 mL). The mixture was stirred at 20 °C for 22 h. The mixture was concentrated under reduced pressure and EtOAc (50 mL) was added. The mixture was stirred at 0 °C for a further 2 h. The resulting solid was filtered off, washed with EtOAc several times, and dried 30 to give crude 19 (172 mg, 96%). LC-MS analysis showed the crude product contained 85% 19 and 13% JV-(2-chloroethyl)-l-(chloromethyl)-3-[5-(2-morpholinoethoxy)-lH-indole-2-carbonyl]-5-nitro-1,2-dihydro- 3i i-benzo[c]nidole-7-carboxamide. The crude product was dissolved in the minimum amount of DMSO (3 mL) and CH2C12 (20 mL) was added slowly. The resulting mixture was stirred at 20 °C for 1 h. The precipitated solid was filtered off, washed repeatedly with CH2C12 and dried to 35 give pure 19 (153 mg, 85%) as a yellowish orange hydrochloride salt (98.1% pure by HPLC), mp 225 59 Received at IPONZ on 14 December 2010 °C (dec); ]H NMR [(CD3)2SO] S 11.80 (poorly resolved d, J = 1.6 Hz, exchangeable with D20, 1 H), ca. 12.0-9.7 (v br s, exchangeable with D20, ca. 3 FI), 9.17 (s, 1 H), 8.97 (t, J ~ 5.5 Hz, exchangeable with D20, 1 H), 8.84 (poorly resolved dj = 1.3 Hz, 1 H), 8.32 (d,/ = 8.8 Hz, 1 H), 8.16 (ddj = 8.8, 1.5 Hz, 1 H), 7.46 (dj = 8.9 Hz, 1 H), 7.27 (poorly resolved dJ = 2.3 Hz, 1 H), 7.22 (poorly 5 resolved dj = 1.6 Hz, collapses to s after D20 exchange, 1 H), 4.96 (br tJ = 10.2 Hz, 1 H), 4.75-4.58 (m, 2 H), 4.41 (t ,J — 4.9 Hz, 2 H), 4.20-3.77 (m, 8 H), 3.64-3.12 (m, partially obscured by water peak, revealed after D20 exchange, 8 H). Anal. (C31H33ClN5O10HCl'H2O) Calc: C, 49.22; H, 4.80; N, 9.26. Found: C, 49.14; H, 4.83; N, 9.17.
Example 15. (£)-l-(Chloromethyl)-At(2-hydroxyethyl)-3-{3-[4-(2- morpholinoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3//-benzo[e]indole-7-carboxamide hydrochloride (6) (Scheme G). To a stirred mixture of indoline 137 (80 mg, 0.17 mmol) and acid 129 (65 mg, 0.21 mmol) in DMA (4 mL) was added EDCI.HCl (132 mg, 0.69 mmol) and anhydrous TsOH (6.0 mg, 0.034 mmol). The mixture was stirred at 20 °C for 5 h and another batch of acid 129 (65 mg, 0.21 mmol), EDCI.HCl (132 mg, 0.69 mmol) and anhydrous TsOH (6.0 mg, 0.034 mmol) were added. After a total reaction time of 21 h the reaction flask was cooled in an ice-bath and cold aqueous NaHCO, (5%, 8 mL) was added, followed by water (8 mL). After stirring at 0 °C for 10 min the precipitated solid was filtered off, washed with cold water several times, and dried to give (E) -N- [2-(fe^-butyldimethylsilyloxy) ethyl] -1 - (chloromethyl)-3- { 3 - [4- (2- morpholinoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3H-benzo[i?]indole-7-carboxamide (141) as a yellowish orange solid (121 mg, 97%): mp 164-165 °C; *H NMR (CDC1,) 8 9.35 (br s, 1 H), 8.75 (poorly resolved d J = 1.3 Hz, 1 H), 8.12 (ddj = 8.8, 1.6 Hz, 1 H), 7.93-7.86 (m, 2 FI), 7.58 (dj = 8.8 Hz, 2 H), 6.95 (dj = 8.8 Hz, 2 H), 6.78-6.69 (m, 2 H), 4.63 (ddj - 10.8, 2.4 Hz, 1 H), 4.53 (tJ = 9.7 Hz, 1 H), 4.33-4.25 (m, 1 H), 4.17 (tj = 5.7 Hz, 2 H), 3.97 (ddJ = 10.4, 3.2 Hz, 1 H), 3.85 (t, J = 5.2 Hz, 2 H), 3.75 (poorly resolved t J = 4.7 Hz, 4 H), 3.69-3.56 (m, 3 H), 2.83 (t J = 5.7 Hz, 2 H), 2.59 (poorly resolved t J = 4.7 Hz, 4 H), 0.93 (s, 9 H), 0.11 (s, 6 H). Anal. (C37FI47ClN407Si) Calc: C, 61.44; H, 6.55; N, 7.75. Found: C, 61.31; H, 6.66; N, 7.99.
To a stirred solution of silyl ether 141 (97 mg, 0.134 mmol) in CH2C12 (12 mL) was added HCl 30 (1.25M in MeOH, 1.0 mL). The mixture was stirred at 20 °C for a further 2 h. EtOAc was added and the mixture was stirred for another 30 min. The resulting solid was filtered off, washed with EtOAc several times, and dried to give 6 as a yellowish orange solid (87 mg, 100%): mp 239-241 °C (dec.); 'H NMR [(CD3)2SO] 8 10.81 (br s, 1 H), 9.22 (s, 1 H), 8.82-8.74 (m, 2 H), 8.24 (dj = 8.8 Hz, 1 H), 8.13 (ddj = 8.8, 1.5 Hz, 1 H), 7.84 (dj= 8.7 Hz, 2 H), 7.74 (dj= 15.3 Hz, 1 H), 7.18-7.05 35 (m, 3 H), 4.72-4.58 (m, 3 H), 4.48 (br s, 2 H), 4.09 (br d J = 4.2 Hz, 2 H), 3.98 (br d J = 12.0, 2 H), 60 Received at IPONZ on 14 December 2010 3.81 (br t J — 11.7 Hz, 2 H), 3.66-3.47 (m, 5 H), 3.40 (qj — 5.9 Hz, 2 H, partially obscured by water peak), 3.26 (br s, 4 H, partially obscured by water peak, revealed on D20 exchange). Anal. (C31H33ClN407-HCrH20) Calc: C, 56.11; H, 5.47; N, 8.44. Found: C, 56.08; H, 5.42; N, 8.44.
Example 16. (£)-2-[l-(Chloromethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3/£benzo[e]indoIe-7-carboxamido] ethyl dihydrogen phosphate hydrochloride (20) (Scheme G). To a stirred mixture of indoline 139 (176 mg, 0.33 mmol) and acid 129 (123 mg, 0.39 mmol) in DMA (4 mL) was added EDCI.HCl (250 mg, 1.30 mmol) and anhydrous TsOH (10,5 mg, 0.06 mmol). The mixture was stirred at 20 °C for 4 h and another batch of acid 129 (123 mg, 10 0.39 mmol), EDCI.HCl (250 mg, 1.30 mmol) and anhydrous TsOH (10,5 mg, 0.06 mmol) were added. After a total reaction time of 21 h the reaction flask was cooled in an ice-bath and cold aqueous NaHC03 (5%, 12 mL) was added, followed by water (12 mL). After stirring at 0 °C for 10 min the precipitated solid was filtered off, washed with cold water several times, and dried to give (E)-dWm'-butyl 2-[l-(chIoromethyl)-3-{3~[4-(2-morphoIinoethoxy)phenyl]acryloyl}-5-nitro-l,2-15 dihydro-3 H-benzo|<?]inc]ole-7-carboxamido) ethy] phosphate (142) as a yellowish orange solid (211 mg, 81%): mp 128-131 °C; 1H NMR (CDC13) 8 9.30 (br s, 1 H), 8.90 (poorly resolved d J = 1.4 Hz, 1 H), 8.17 (ddj - 8.7, 1.4 Hz, 1 H), 8.01 (tJ = 4.6 Hz, 1 FI), 7.94-7.86 (m, 2 H), 7.58 (dJ = 8.7 FIz, 2 H), 6.96 (dj= 8.7 Hz, 2 FI), 6.76 (dJ = 15.2 Hz, 1 H), 4.62 (ddj= 10.8, 2.1 Hz, 1 H), 4.52 (br tJ= 9.7 Hz, 1 FI), 4.32-4.20 (m, 3 H), 4.17 (tJ = 5.7 FIz, 2 H), 3.96 (ddj = 11.4, 3.1 Hz, 1 H), 20 3.81 (qj — 4.6 FIz, 2 H), 3.74 (poorly resolved tJ = 4.6 Hz, 4 H), 3.59 (br t,J — 10.6 Hz, 1 H), 2.84 (t J = 5.7 FIz, 2 H), 2.60 (poorly resolved t J = 4.5 Hz, 4 H), 1.50 (s, 18 H). Anal. (C39Ha,ClN4O10P-y2H2O) Calc: C, 57.81; H, 6.34; N, 6.91. Found: C, 57.51; H, 6.10; N, 6.95.
To a stirred solution of phosphate ester 142 (190 mg, 0.237 mmol) in CH2C12 (10 mL) was added 25 HCl (saturated solution in CH2C12, 10 mL). The mixture was stirred at 20 °C for a further 18 h. The mixture was concentrated under reduced pressure and EtOAc was added. The mixture was stirred at 0 °C for another 2 h. The resulting solid was filtered off, washed with EtOAc several times, and dried to give 20 as a yellowish orange solid (147 mg, 86%): mp 229 °C (dec.); 11 [ NMR [(CD3)2SO] S ca.10.6 (v br s, 1 FI), 9.23 (br s, 1 H), 8.97 (poorly resolved t J = 4.7 Hz, 1 H), 8.82 (poorly resolved 30 dj = 0.9 Hz, 1 H), 8.26 (dj = 8.8 Hz, 1 H), 8.14 (br dJ = 8.7 Hz, 1 H), 7.83 (dJ = 8.5 Hz, 2 H), 7.73 (dJ = 15.3 Hz, 1 H), 7.12 (dj = 15.3 Hz, 1 H), 7.08 (dj - 8.5 Hz, 2 H), 4.72-4.56 (m, 3 H), 4.39 (br s, 2 H), 4.12-3.95 (m, 4 H), 3,80 (br s, 4 H), 3.55 (q J = 5.6 Hz, 2 H), 3.37 (br s, 2 H, partially obscured by water peak, revealed on D20 exchange), 3.15 (br s, 4 H, partially obscured by water peak, revealed on DaO exchange). Anal. (C3lH34ClN4O10P,HCry4H2O) Calc: C, 51.07; H, 4.89; 35 N, 7.68. Found: C, 51.00; H, 4.90; N, 7.68. 61 Received at IPONZ on 14 December 2010 Example 17. (5)-l-(Chloromethyl)-7V-(2-hydroxyethyl)-3- [5-(2-morpholinoethoxy)-l/£-indole-2-carb onyl] -5-nitro- l,2-dihydro-3/£-b enz o [e] indole-7-sulfonamide hydro chloride (7) (Scheme H and I). An ice-cold solution of amine 114 (3.39 g, 13.4 mmol) and DIPEA (2.10 mL, 5 11.8 mmol) in THF (17 mL) was added to solution of sulfonyl chloride 107 (5.12 g, 11.2 mmol) in THF (50 mL) at 0 °C. After the addition was complete the mixture was stirred at 0 °C for a further 20 min. Ice-watet (400 mL) was added and the mixture was stirred for 15 min. The precipitated solid was filtered off, washed with water, and dried to give diffef-butyl) 2-[l-(chloromethyI)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3i^-benzo[tf]indole-7-sulfonamido]ethyl phosphate (143) as a yellow 10 solid (7.19 g, 95%): mp 162 °C (dec.); 'H NMR (CDC13) 5 9.23 (s, 1 FI), 9.01 (d J = 1.5 Hz, 1 H), 8.10 (ddj = 8.8,1.5 Hz, 1 H), 8.00 (dj = 8.8 Hz, 1 H), 6.10 (tj= 5.5 Hz, 1 H), 4.72 (dj= 11.5 Hz, 1 H), 4.57 (ddj = 11.4, 8.7 Hz, 1 H), 4.41-4.31 (m, 1 H), 4.13-4.03 (m, 2 H), 3.94 (ddj = 11.6, 3.4 Hz, 1 H), 3.71 (dd, J — 11.6, 7.9 Hz, 1 H), 3.33 (q J = 5.5 Hz, 2 H), 1.45 and 1.46 (2 s, 18 H). Anal. (C^HjjClFjNjOgPS'VVTHF) Calc: C, 45.13; FI, 4.95; N, 6.07. Found: C, 45.19; H, 4.97; N, 6.21.
Compound 143 (3.68 g) was resolved by preparative chiral HPLC (Daicel Chiralpak IA 250 x 21 mm column, EtOH:hexane 40:60, 7 mL/min, run time 27 min, a 1.30). The sample was dissolved in a mixture of MeOH and EtOH and then diluted with hexane, and 55-60 mg portions were resolved per injection. The product-containing fractions were combined and evaporated to give (R) d\lerl-20 butyl 2-(l-(chloromethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3H-benzo[(?]indole-7- sulfonamido)ethyl phosphate (144) as a yellowish orange solid (1.425 g, 38.7%): mp 153-154 °C (EtOAc/petroleum ether); [«]D20 +28° (c 1.0, CHCL,); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOH:hexane 40:60, 0.5 mL/min, Rr 13.54 min); 1H NMR (CDCl,) identical to that described for 143. Anal. (C25H32C1F3N309PS'0.1 hexane) Calc: C, 45.04; H, 4.93; N, 6.16. Found: C, 25 45.09; H, 4.94; N, 6.28. The structure of phosphate 144 was confirmed by X-ray crystallography, see Figure 1. and (j)-di-/i?^-butyl 2-[l-(chloromethyI)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3.H-benzo[V]indole-7-sulfonamido]ethyl phosphate (145) as a yellowish orange solid (1.423 g, 38.7%): mp 154-155 °C 30 (EtOAc/petroleum ether); [a]D20 -28° (c 1.0, CHC13); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOH:hexane 40:60, 0.5 mL/min, RT 17.47 min); 1H NMR (CDC13) identical to that described for 143. Anal. (C25H32ClF3N3OgPS) Calc: C, 44.55; H, 4.79; N, 6.23. Found: C, 44.49; H, 4.93; N, 6.19. The structure of phosphate 145 was confirmed by X-ray crystallography, see Figure 2.
Received at IPONZ on 14 December 2010 Cold THF (32 mL) was added to solid trifluroacetamide 144 (1.38 g, 2.05 mmol) in a flask immersed in an ice-bath. To this stirred cold mixture was added solid cesium carbonate (1.33 g, 4.1 mmol) followed by slow addition of cold MeOH (8 mL). The mixture was stirred for 10 min and then partitioned between EtOAc and ice-water. The EtOAc layer was separated and the aqueous layer 5 was extracted with EtOAc once more. The combined EtOAc extracts was dried (Na2S04) and evaporated to give (R)-di-^n'-butyl 2-[l-(chloromethyl)-5-nitro-l,2-dihydro-3 H-benzo [?jindolc 7-sulfonamido]ethyl phosphate (146) as a red foamy solid (1.19 g, 100%): mp 94-96 °C (EtOAc); [a]n20 -27° (c 1.0, CFICL); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOH:hexane 40:60, 0.5 mL/min, RT 14.92 min); 'H NMR (CDClj) 8 8.92 (poorly resolved dj = 1.5 Hz, 1 H), 7.91 (dd,/ = 10 8.9, 1.8 Hz, 1 H), 7.79 (ddj = 8.9, 0.33 Hz, 1 H), 7.74 (s, 1 H), 5.71 (tj = 5.8 Hz, 1 H), 4.40 (s, 1 H), 4.18-3.91 (m, 5 H), 3.78 (ddj = 11.2, 3.5 Hz, 1 H), 3.59 (ddj = 11.1,10.0 Hz, 1 H), 3.35-3.24 (m, 2 H), 1.46 and 1.45 (2 s, 18 FI). Anal. (C^H^ClN^OgPS) Calc: C, 47.79; H, 5.75; N, 7.27. Found: C, 47.59; H, 5.79; N, 6.96.
Trifluoroacetamide 145 (1.38 g, 2.05 mmol) was deprotected to the corresponding indoline by the same procedure to give (J)-di-&n?-butyl 2-[l-(chloromethyl)-5-nitro-l,2-dihydro-3H-benzo[fjindole-7-sulfonamido]ethyl phosphate (147) as a red foamy solid (1.19 g, 100%): mp 89-92 °C (EtOAc); [ot]D20 +29° (c 1.0, CFIClj); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOFFhexane 40:60, 0.5 mL/min, RT 17.78 min); lH NMR (CDC13) identical to that described for 165. Anal. 20 (C^FI^ClN-OgPS) Calc: C, 47.79; FI, 5.75; N, 7.27. Found: C, 48.23; H, 5.79; N, 7.01.
Compound 115 (4.26 g) was resolved by preparative chiral HPLC (Daicel Chiralpak IA 250 x 21 mm column, EtOH:hexane 40:60, 7 mL/min, run time 24 min, a 1.18). The sample was dissolved in EtOH:hexane (40:60) and 80 mg portions were resolved per injection. The product-containing 25 fractions were combined and evaporated to give R-enantiomer 146 (1.91 g, 44.8%), ee 100% (analytical FIPLC conditions as described above); and ^-enantiomer 147 (1.93 g, 45.3%), ee 100% (analytical HPLC conditions as described above).
A solution of 147 (174 mg, 0.30 mmol) in CH2C12 (15 mL) was treated with TFA (1.5 mL) at 20 °C 30 for 30 min. The mixture was evaporated under reduced pressure to give a thick yellow oil. To the oil was added water (225 mL) and acid phosphatase (wheat germ, 0.5 units/mg, 300 mg). The mixture (pH ca. 5.5) was stirred at 20 °C for 23 h and then extracted with EtOAc (2 x 300 mL). The combined extracts were washed with water, dried (Na2S04), and evaporated under reduced pressure at 35 °C (bath temperature) to give (j)-l-(chloromethyI)-iV-(2-hydroxyethyl)-5-nitro-l,2-dihydro-35 3H-benzo[e]indole-7-sulfonamide (148) as a red solid (105 mg, 91%): mp 170-171 °C 63 Received at IPONZ on 14 December 2010 (EtOAc/MeOH); [a]D20 +90.5° (c 0.22, EtOAc); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOH:hexane 60:40, 0.5 mL/min, RT 20.88 min); 'Et NMR [(CD3)2SO] 8 8.57 (poorly resolved d J = 1.6 Hz, 1 H), 8.03 (dJ - 8.9 Hz, 1 H), 7.80 (ddj - 8.9, 1.8 Hz, 1 H), 7.77 (s, 1 H), 7.69 (tj = 5.9 Hz, 1 H), 6.73 (s, 1 H), 4.63 (tJ = 5.6 Hz, 1 H), 4.29-4.20 (m, 1 H), 3.96-3.71 (m, 4 5 H), 3.37 (br q J = 6.1 Hz, 2 H), 2.81 (br q J = 6.2 Hz, 2 H). Anal. (C15H16C1N305S) Calc: C, 46.70; H, 4.18; N, 10.89. Found: C, 46.88; H, 4.27; N, 10.64.
To a stirred solution of 148 (107 mg, 0.32 mmol) in DMF (5 mL) was added tert-buuyldirnef.hy lsi ly 1 chloride (84 mg, 0.55 mmol), followed by DIPEA (0.10 mL, 0.58 mmol). The mixture was stirred at 10 20 °C for 45 min and then partitioned between EtOAc and cold water. The ethyl acetate layer was separated and washed with cold 2% Na2COs and water. The organic layer was dried (Na2S04) evaporated to give (J)-IV-[2-(/^4)utyldimethylsilyloxy)ethyl]-l-(chloromethyl)-5-nitro-l,2-dihydro-3il-benzo [ <?| indole-7-sul fonatmide (149) as a red solid (134 mg, 100%): 'FI NMR (CDC13) identical to that described for 112.
To a stirred mixture of 149 (134 mg, 0.27 mmol) and acid 122 (132 mg, 0.40 mmol) in DMA (5 mL) was added EDCI.HCl (308 mg, 1.61 mmol) and anhydrous TsOH (14.0 mg, 0.08 mmol). The mixture was stirred at room temperature for 2 h and another batch of acid 122 (28 mg, 0.084 mmol), EDCI.HCl (51 mg, 0.27 mmol), and anhydrous TsOH (2.3 mg, 0.013 mmol) was added. The 20 reaction mixture was stirred for a further 1 h 30 min, then cooled in an ice bath. Cold aqueous NaFIC03 (5%, 10 mL) was added, followed by cold water (10 mL). The precipitated solid was filtered off, washed with water then hexane, and dried to give (S)-N'[2-(tert-butyldimethylsilyloxy)ethyl] -1 - (chloromethyl)-3-[5 - (2-morpholinoethoxy) -1 H-indole-2-carbonyl]-5 -nitro-l,2-dihydro-3H-benzo[e]indole-7-sulfonamide (150) as a yellowish orange solid (198 mg, 95%): 25 mp 179-181 °C; [a]D2u +22.5° (c 0.71, CFIC1,); 'H NMR [(CDC1J 8 9.36 (s, 1 H), 9.33 (br s, 1 H), 9.02 (poorly resolved dj = 1.4 Hz, 1 H), 8.05 (ddj = 8.8, 1.6 Hz, 1 H), 8.00 (dJ = 8.8 Hz, 1 H), 7.39 (dj = 8.9 Hz, 1 H), 7.14 (poorly resolved dJ = 2.0 Hz, 1 H), 7.12-7.03 (m, 2 H), 5.01 (tJ = 5.8 Hz, 1 H), 4.93 (dd J = 10.8, 2.1 Hz, 1 H), 4.83 (br t J = 9.8 Hz, 1 H), 4.41-4.30 (m, 1 H), 4.18 (t, } = 5.7 Hz, 1 H), 3.96 (ddj =11.5, 3.2 Hz, 1 H), 3.83-3.60 (m, 7 H), 3.23-3.14 (m, 2 H), 2.86 (tJ = 30 5.7 Hz, 2 H), 2.68-2.56 (m, 4 H), 0.84 (s, 9 H), 0.005 and 0.002 (2 s, partially obscured by TMS, 6 H). Anal. (C36H46ClN508SSi0.3hexane) Calc: C, 56.88; H, 6.34; N, 8.77. Found: C, 56.71; H, 6.47; N, 8.66.
To a filtered solution of 150 (189 mg, 0.245 mmol) in CH2C12 (15 mL) was added HCl (1.25M in 35 MeOH, 1.5 mL) and the mixture was stirred at 20 °C for 1 h 15 min. The mixture was diluted with 64 Received at IPONZ on 14 December 2010 EtOAc (100 mL) and the precipitated solid was filtered off, washed with EtOAc several times, and dried to give 7 as a yellowish orange hydrochloride salt (144 mg, 85%): mp 220 °C (dec.); [ot]D2u +32.0° (c 0.17, MeOH); 1H NMR [(CD3)2SO] 5 11.82 (br s, exchangeable with D20, 1 H), 10.63 (br s, exchangeable with D20, 1 H), 9.28 (s, 1 H), 8.86 (poorly resolved d J = 1.6 Hz, 1 EI), 8.44 (d J — 5 8.9 Hz, 1 H), 8.03 (dd, / = 8.9, 1.7 Hz, 1 H), 7.94 (t J = 5.9 Hz, exchangeable with DzO, 1 H), 7.47 (dJ = 8.9 Hz, 1 H), 7.28 (poorly resolved d,J— 2.2 Hz, 1 H), 7.24 (poorly resolved d,/ = 1.7 Hz, collapses to s after D20 exchange, 1 H), 7.05 (dd, / = 8.9, 2.4 Hz, 1 H), 4.97 (br t J = 10.2 Hz, 1 H), 4.77-4.61 (m, 2 H), 4.49-4.39 (m, 2 H), 4.22-4.08 (m, 2H), 4.06-3.49 (m, 8 H), 3.47-3.17 (m, partially obscured by water peak, revealed after DaO exchange, 4 H), 2.87 (qJ = 6.1 Hz, collapses to t after 10 D20 exchange, 2 H), one H not observed. Anal. (C30H32C1N5OsS HC1'H20) Calc: C, 50,57; H, 4,95; N, 9.83. Found: C, 50.55; FI, 5.19; N, 9.98.
Example 18. (5)-5-Aniino-l-(ctiloromcthyl)-/V-(2-hydroxyethyl)-3- [5-(2-morpholinoethoxy)-l//-itidolc-2-carbonyl]-l,2-dibydro-3/^-benzo[e']iiidole-7-suIfoiiamide hydrochloride (14) (Scheme I). To a solution of nitro compound 7 (78 mg, 0.112 mmol) in acetone (6 mL) and water (4 mL) stirred under nitrogen at 20 °C was added ammonium chloride (312 mg, 5.8 mmol) and Zn powder (234 mg, 3.6 mmol). The heterogeneous mixture was stirred at 20 °C for 1 h 30 min. Sufficient cold aqueous Na,CO, (2%) was added to generate pFI 8-9 and then the product was extracted with CH2C12 (15 x lOOmL). The combined CH2C12 extracts were washed with water, dried 20 (Na2S04), and concentrated under reduced pressure to ca. 10 mL in volume. HCl (1.25M in MeOH, 1.0 mL) was added followed by EtOAc (60 mL). The mixture was left to stand at 5 °C overnight. The precipitated solid was filtered off, washed with EtOAc, and dried to give 14 as greenish beige solid (60 mg, 81%): mp 230 °C (dec.); [a]D20 +17.6° (c 0.17, MeOH); 1H NMR j(CD3)2SO] 8 11.67 (poorly resolved d J = 1.7 Hz, exchangeable with D20, 1 H), 10.70 (br s, exchangeable with DzO, 1 25 H), 8.55 (poorly resolved d J = 1.6 FIz, 1 H), 7.95 (d J = 8.9 Hz, 1 H), 7.86 (s, 1 FI), 7.76 (ddj = 8.9, 1.7 Hz, 1 FI), 7.49 (poorly resolved t J — 5.7 Hz, exchangeable with D20, 1 H), 7.45 (d J = 8.9 Hz, 1 H), 7.27 (poorly resolved d J = 2.3 Hz, 1 H), 7.12 (poorly resolved d J = 1.7 Hz, collapses to s after D20 exchange, 1 FI), 7.01 (ddj = 8.9, 2.4 Hz, 1 H), 4.77 (ddj = 10.7, 9.1 FIz, 1 FI), 4.53 (dd, / = 10.9, 1.7 Hz, 1 H), 4.43 (poorly resolved t J = 4.9 Hz, 2 H), 4.24-4.16 (m, 1 H), 4.06-3.61 (m, 30 partially obscured by water peak, revealed after D20 exchange, 6 H), 3.58-3.06 (m, partially obscured by water peak, revealed after D20 exchange, 8 H), 2.84 (br q J = 6.0 Hz, collapses to t after DaO exchange J = 6.0 FIz, 2 H). HRMS (FAB) calc for C3aH35:!5ClN506S (MFI+) 628.1997, found 628.1996. For C30H3537ClN5O6S (MH+) 630.1967, found 630.1974. 65 Received at IPONZ on 14 December 2010 Example 19. (Chloromethyl)-3-[5-(2-morpholinoethoxy)-1 //-indoIe-2-carhonyl]-5 - nitro-l,2-dihydro-3/f-benzo [e]indoIe-7-sulfonamido] ethyl dihydrogen phosphate trifluoroacetate (21) (Scheme I). To a stirred mixture of 147 (1.93 g, 3.35 mmol) and acid 122 (1.38 g, 4.22 mmol) in DMA (70 mL) was added EDCI.HCl (1.29 g, 6.70 mmol) and anhydrous 5 TsOH (117 mg, 0.67 mmol). The mixture was stirred at 20 °C for 2 h and another batch of EDCI.HCl (1.29 g, 6.70 mmol) and anhydrous TsOH (59 mg, 0.34 mmol) was added. The reaction mixture was stirred for a further 3 h, then cooled in an ice bath. Cold aqueous NaHCO, (5%, 140 mL) was added, followed by cold water (140 mL). The precipitated solid was filtered off, washed with water, and dried to give (ij-dl-/<?r/-butyl 2-[l-(chloromethyl)-3-[5-(2-morpholinoethoxy)-lH-10 indole-2-carbonyl]-5-nitro-l,2-dihydro-3H-benzo[f]indole-7-sulfonamido]ethyl phosphate (151) as a yellowish orange solid (2.70 g, 95%): mp 219 °C; [a]D20 +20.1° (c 0.70, CHCt,); 'H NMR [CDC13] 5 9.37 (br s, 1 H), 9.33 (s, 1 H), 8.99 (poorly resolved dj = 1.5 Hz, 1 H), 8.05 (ddj - 8.8, 1.7 Hz, 1 H), 7.98 (dj = 8.8 Hz, 1 H), 7.38 (dj - 8.9 Hz, 1 H), 7.14 (poorly resolved dJ = 2.2 Hz, 1 H), 7.10-7.06 (m, 2 H), 6.10-5.97 (m, 1 H), 4.92 (ddj = 10.8, 2.2 EIz, 1 H), 4.82 (br tJ = 9.8 Hz, 1 EI), 15 4.39-4.30 (m, 1 H), 4.18 (tj = 5.7 EIz, 2 H), 4.13-4.03 (m, 2 H), 3.96 (ddj = 11.5, 3.3 Hz, 1 H), 3.76 (poorly resolved tJ= 4.7 Hz, 4 H), 3.66 (ddj = 11.5, 9.1 Hz, 1 H), 3.39-3.29 (m, 2 H), 2.86 (t, J — 5.7 Hz, 2 H), 2.62 (poorly resolved t J = 4.5 Hz, 4 H), 1.47 and 1.46 (2 s, 18 H). Anal. (C^H^ONjOnPS-VaHaO) Calc: C, 53.11; H, 5.87; N, 8.15. Found: C, 53.03; H, 5.90; N, 8.20.
TFA (5 mL, 63 mmol) was added to a filtered solution of phosphate ester 151 (2.70 g, 3.18 mmol) in CH2C12 (50 mL) and the resulting solution was stirred at 20 °C for 3 h. EtOAc (500 mL) was added, the mixture was stirred for a further 3 h, and then left to stand at 5 °C overnight. The precipitated solid was filtered off, washed with EtOAc several times and dried to give 21 as a yellowish orange solid (2.63 g, 97%): mp 210 °C (dec.); [a]D20 +19.7° (c 0.20, DMSO); ]H NMR [(CD3)2SO] 5 11.75 25 (br s, 1 H), 9.27 (s, 1 FI), 8.86 (d J = 1.5 Hz, 1 H), 8.42 (d J = 8.9 Hz, 1 H), 8.23 (br s, 1 H), 8.02 (ddj - 8.9, 1.5 Hz, 1 H), 7.41 (dj = 8.9 FIz, 1 H), 7.21 (br s, 2 FI), 6.96 (ddj = 8.9, 1.3 Hz, 1 H), 4.94 (br t J = 10.1 Hz, 1 H), 4.76-4.57 (m, 2 H), 4.22 (poorly resolved t,J= 5.2 Hz, 2 H), 4.19-4.05 (m, 2 H), 3.86-3.66 (m, partially obscured by water peak, 6 H), 3.14-2.78 (m, partially obscured by water peak, 8 H), 3 H not observed, obscured by water peak 8 3.5-3.3. Anal. 30 (C30H33ClNsOtlPS'07CF3C02H-Y2H20) Calc: C, 45.61; H, 4.23; N, 8.47; CI, 4.29; F, 4.83. Found: C, 46.00; H, 4.35; N, 8.04; CI, 4.20; F, 4.67.
Example 20. (5,JE)-2-[l-(Chloromethyl)-3~{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-1,2-dihydro-3i/-benzo [ e] indole-7-sulfonamido] ethyl dihydrogen phosphate 35 trifluoroacetate (22) (Scheme J). To a stirred mixture of indoline 147 (116 mg, 0.20 mmol) and 66 Received at IPONZ on 14 December 2010 acid 129 (79.0 mg, 0.25 mmol) in DMA (4 mL) was added EDCI.HCl (154 mg, 0.80 mmol) and anhydrous TsOH (7.0 mg, 0.04 mmol). The mixture was stirred at 20 °C for 1 h 30 min and another batch of acid 129 (79.0 mg, 0.25 mmol), EDCI.HCl (154 mg, 0.80 mmol) and anhydrous TsOH (7.0 mg, 0.04 mmol) were added. After a total reaction time of 18 h, the reaction flask was immersed in 5 an ice-bath and cold aqueous NaHCO, (5%, 8 mL) was added, followed by water (8 mL). The mixture was stirred at 0 °C for 10 min, then the precipitated solid was filtered off, washed with water then petroleum ether, and dried to give (i',L)-di-/cr/-butyI 2-[l-(chloromelhyi) 3 {3-[4-(2-morphoHnoethoxy)phenyl]acryloyl}-5-nitro-l,2-dihydro-3H-benzo[(?]indole-7-sulfonamido]ethyl phosphate (152) as a yellowish orange solid (155 mg, 93%): mp 255 °C (dec.); [a]D20 -6.5° (c 0.62, 10 CHCL); ee 100% (Daicel Chiralpak IA 150 x 4.6 mm column, EtOH:acetomtrile:diethylamine :65:0.1, 0.6 mL/min, RT 13.82 min); aH NMR (CDC13) 5 9.41 (br s, 1 H), 8.97 (poorly resolved dJ = 1.5 Hz, 1 H), 8.04 (ddj = 8.9, 1.6 Hz, 1 EI), 7.94 (dj = 8.9 Hz, 1 H), 7.90 (dj = 15.2 Hz, 1 H), 7.57 (dJ = 8.7 Hz, 2 H), 6.96 (dj= 8.7 Hz, 2 H), 6.74 (dj = 15.2 Hz, 1 H), 5.92 (tJ = 5.4 Hz, 1 H), 4.63 (br ddj = 10.7, 2.3 Hz, 1 EI), 4.55 (br t J - 9.7 Hz, 1 H), 4.34-4.24 (m, 1 H), 4.17 (t J = 15 5.7 Hz, 2 H), 4.13-4.02 (m, 2 H), 3.95 (br ddj = 11.4, 3.2 Hz, 1 H), 3.75 (poorly resolved tJ = 4.7 Hz, 4 H), 3.62 (ddj = 11.4, 9.4 Hz, 1 H), 3.32 (poorly resolved qJ = 5.0 Hz, 2 H), 2.83 (tj = 5.7 Hz, 2 H), 2.59 (poorly resolved t J = 4.6 Hz, 4 H), 1.46 and 1.45 (2 s, 18 EI). Anal.
(C3aH50ClN4O11PS) Calc: C, 54.51; H, 6.02; N, 6.69. Found: C, 54.29; H, 5.97; N, 6.71.
TFA (2 mL, 25 mmol) was added to a filtered solution of phosphate ester 152 (149 mg, 0.18 mmol) in CFI2C12 (20 mL) and the resulting solution was stirred at 20 °C for 1 h 30 min. EtOAc (80 mL) was added, the mixture was stirred for a further 2 h, and then left to stand at 5 °C overnight. The precipitated solid was filtered off, washed with EtOAc several times, and dried to give 22 as a yellowish orange solid (105 g, 71%): mp 213 °C (dec.); [a]D20 +38.5° (c 0.21, DMSO); 'FI NMR 25 (CSDSN) 8 ca. 10.6 (v br s, 1 H), 9.81 (br s, 1 FI), 9.45 (poorly resolved d J — 1.4 Hz, 1 H), 8.40 (poorly resolved ddj = 8.8, 1.5 Hz, 1 H), 8.21 (dj = 15.2 FIz, 1 H), 8.14 (dj = 8.8 FIz, 1 H), 7.73 (dJ = 8.7 Hz, 2 H), 7.16 (dJ = 15.2 Hz, 1 H), 7.10 (dj = 8.7 Hz, 2 H), 4.80 (poorly resolved ddj = 10.8, 2.5 Hz, 1 H), 4.65 (br t J = 10.0 Hz, 1 H), 4.58-4.42 (m, 3 H), 4.22-4.12 (m, 3H), 4.00 (br ddj = 11.3, 8.3 FIz, 1 H), 3.78-3.66 (m, 6 H), 2.75 (tJ = 5.7 Hz, 2 H), 2.57-2.48 (m, 4 H), 3 H not 30 observed, obscured by water peak. Anal. (C30H34ClN4OnPS'0.8CF3C02H) Calc: C, 46.50; H, 4.30; N, 6.86. Found: C, 46.50; H, 4.64; N, 7.00.
Example 21. l-(Bromomethyl)-At(2-hydroxyethyl)-3- [5-(2-morpholinoethoxy)-l//-incloIc-2-carbonyl]-5-nitro-l)2-dihydro-3J££benzo[e]indoie-7-sulfonatmdc (8) (Scheme K and L). A solution of Nal (7.19 g, 47.9 mmol) in 2-butanone (30 mL) was heated at 85 °C for 1 h. Indoline 105 67 Received at IPONZ on 14 December 2010 (5.0 g, 16.0 mmol) was added and the mixture was heated at 85 °C for a further 17 h. EtOAc and water were added and the organic layer was separated. The organic layer was washed successively with aqueous sodium disulfite (10%), water, and brine, and then dried (Na2S04) and evaporated. The residue was dissolved in EtOAc and the solution was diluted with petroleum ether to give 1-5 (iodomethyl)-3-(ttifluoroacetyl)-l^-dihydro-3H-benzo[e]indole (153) as a cream powder (4.91 g, 76%): mp 132-135 °C; *H NMR (CDC13) 8 8.42 (dj = 9.0 Hz, 1 H), 7.92 (d ,J= 7.9 Hz, 1 FI), 7.89 (dJ = 8.9 Hz, 1 H), 7.77 (dj= 8.4 Hz, 1 H), 7.58 (td J = 8.1,1.1 Hz, 1 H), 7.48 (tdj= 8.1, 1.1 Hz, 1 H), 4.49 (dJ = 11.5 Hz, 1 H), 4.43 (ddj = 11.5, 8.2 Hz, 1 H), 4.21 (ttj = 9.8, 2.3 Hz, 1 H), 3.65 (ddj = 10.5, 2.3 Hz, 1 H), 3.23 (t J - 10.2 Hz, 1 H). Anal. (C^H^INO) Calc: C, 44.47; H, 10 2.74; N, 3.46. Found: C, 44.74; H, 3.04; N, 3.41.
A solution of iodide 153 (3.6 g, 8.89 mmol) and AgOMs (10.8 g, 53.3 mmol) in CH3CN (50 mL) was stirred in the dark for 24 h. The CH3CN was removed under reduced pressure, EtOAc was added to the residue, and the resulting mixture was filtered through Celite. The filtrate was evaporated and the 15 crude product was recrystallised (EtOAc/petroleum ether) to give (3-trifluoroacetyl-l,2-dihydro-3H-benzo[c]indol-1 -yl)mechyl methanesulfonate 154 as colourless crystals (3.15 g, 95%): mp 122-124 °C; ]H NMR (CDC13) 8 8.43 (d J = 9.0 Hz, 1 H), 7.91 (dj= 7.8 Hz, 1 H), 7.88 (dj= 8.8 Hz, 1 H), 7.83 (ddj = 8.3, 0.5 Hz, 1 H), 7.61 (tdj = 8.2, 1.2 Hz, 1 H), 7.51 (tdj = 8.1, 1.1 Hz, 1 H), 4.65 (ddj = 3.8, 0.6 Hz, 1 H), 4.59 (dtj= 11.5, 1.4 Hz, 1 H), 4.42 (ddj = 11.5, 8.2 Hz, 1 FI), 4.29 (tdj 20 = 8.2, 3.4 Hz, 1 H), 4.14 (dd J = 10.6, 9.1 Hz, 1 H), 2.95 (s, 3 H). Anal. (C16H14F3N04S) Calc: C, 51.47; H, 3.78; N, 3.75. Found: C, 51.77; H, 3.82; N, 3.86.
A solution of mesylate 154 (8.9 g, 23.9 mmol) and LiBr (41.5 g, 477 mmol) in THF (100 mL) was stirred in the dark for 36 h. EtOAc was added and the solution was washed with water and brine 25 and then dried (Na2S04) and evaporated. The crude product was recrystallised (MeOH/water) to give l-(bromomethyl)-3-(trifluoroacetyl)-l,2-dihydro-3H-ben2o[£?]indole (155) as colourless crystals (8.1 g, 95%): mp 152-154 °C; 'FI NMR (CDC13) 8 8.43 (dj = 9.0 Hz, 1 H), 7.92 (dJ = 9.1 Hz, 1 H), 7.88 (dJ = 9.3 Hz, 1 H), 7.79 (dj= 8.3 Hz, 1 H), 7.58 (tdj = 8.1, 1.2 Hz, 1 H), 7.49 (tdj = 8.1, 1.1 Hz, 1 FI), 4.62 (dtj= 11.5, 1.4 Hz, 1 H), 4.43 (dd J = 11.4, 8.5 Hz, 1 H), 4.25 (tj= 9.1 Hz, 1 30 H), 3.84 (ddd J = 10.7, 3.2, 0.8 Hz, 1 H), 3.39 (t J = 10.2 Hz, 1 H). Anal. (C15H11BrF3NO) Calc: C, 50.30; H, 3.10; N, 3.91. Found: C, 50.58; H, 3.07; N, 3.92.
A solution of 105 (500 mg, 1.60 mmol) and LiBr (2.19 g, 26.39 mmol) was heated at 85 °C in 2-butanone (15 mL) for 9 days. EtOAc and water were added and the organic layer was separated. The 35 organic layer was washed with water and brine and then dried (Na2S04) and evaporated. The residue Received at IPONZ on 14 December 2010 was purified by flash column chromatography (eluting with petroleum ether:EtOAc 100:0 then 99.5:0.5) to give 155 (333 mg, 55%) contaminated with 105 (37 mg, 10%). Attempted recrystallisation (EtOAc/petroleum ether) failed to remove 105.
A solution of mesylate 154 (1.40 g, 3.75 mmol) in CH2C12 (5 mL) was added dropwise to a solution of CISO,El (1.74 g, 15.0 mmol) in CH2C12 (20 mL) at -78 °C. The stirred mixture was allowed to warm to 15 °C over 12 h producing a grey precipitate. The mixture was then cooled to 0 °C and the minimum quantity of DMF was slowly added to dissolve the precipitate. Oxalyl chloride (1.5 mL) was added dropwise, the mixture was stirred at 0 °C for 1 h, and then poured into ice water. The 10 mixture was extracted with cold EtOAc and the organic layer was washed with cold water, cold brine, and then dried (Na2S04). The solution was filtered through a plug of Celite/silica gel and the filtrate was evaporated. The crude product was dissolved in CH2Cl2and the solution was diluted with 2-Pr20 to precipitate [7-{chlorosii1fonyl)-3-(trifluoroacetyl) 1,2 dihydro 3 FT benzo|i?]indo1 l--yl]methyl methanesulfonate (156) as a white powder (1.64 g, 94%): *H NMR [(CD3)2SO] 5 8.32 (dj= 8.9 Hz, 15 1 H), 8.21 (dJ = 1.4 Hz, 1 H), 8.06 (dj = 9.0 Hz, 1 H), 8.00 (dJ = 8.7 Hz, 1 H), 7.81 (ddj = 8.7, 1.6 Hz, 1 H), 4.58 (ddj = 10.0, 3.2 Hz, 1 H), 4.58-4.54 (m, 1 H), 4.49-4.36 (m, 3 H), 3.05 (s, 3 H). FIRMS (FAB) calc for C16H1335C1F3N06S2 (MH+) m/%-470.9825, found 470.9824. For C16H!337C1F3N06S2 (MFI+) »/? 472.9795, found 472.9801.
A solution of indoline 155 (7.50 g, 21.0 mmol) in CH2C12 (40 mL) was added dropwise to a solution of CISOjH (9.72 g, 83.8 mmol) in CH2C12 (40 mL) at -78 °C. The stirred mixture was allowed to warm to 5 °C over 7 h producing a grey precipitate. The mixture was kept at 5 °C overnight and then cooled to 0 °C. The minimum quantity of DMF was slowly added to dissolve the precipitate and then oxalyl chloride (4 mL) was added dropwise. After stirring at 0 °C for 1 h the mixture was 25 poured into ice water and extracted with cold EtOAc. The organic layer was washed with cold water, cold brine, and then dried (Na2S04). The solution was filtered through a plug of Celite/silica gel and the filtrate was evaporated. The crude product was dissolved in CFI2Cl2and the solution was diluted with i-Pr,0 to precipitate 1 -(bromomcthyl) 3 (trifluoroacctyl) l .2 dihydro 3FT benzo [tfjindole-7-sulfonyl chloride (157) as a white powder (8.88 g, 93%): mp 194-196 °C; 1H NMR 30 (CDC13) 8 8.65 (dj - 7.5 Hz, 1 H), 8.63 (dj = 2.0 Hz, 1 H), 8.11 (dj = 9.0 Hz, 1 H), 8.09 (ddj = 9.0, 2.0 Hz, 1 H), 7.99 (dj= 9.0 Hz, 1 H), 4.67 (dtj= 11.7, 1.6 Hz, 1 H), 4.52 (ddj = 11.4, 8.7 Hz, 1 H), 4.32 (ttj = 8.7, 2.6 Hz, 1 H), 3.80 (ddj = 10.8, 3.3 FIz, 1 FI), 3.47 (ddj = 10.8, 9.1 Hz, 1 H). Anal. (C15H10BrClF3NO3SUl/-Pr2O) Calc: C, 40.13; H, 2.46; N, 3.00. Found: C, 40.17; H, 2.27; N, 2.97. 69 Received at IPONZ on 14 December 2010 A solution of mesylate 156 (80 mg, 0.17 mmol) and LiBr (400 mg, 4.60 mmol) in THF (8 mL) was stirred at 20 °C for 4 days. EtOAc and water were added and the organic layer was separated and then washed with water and brine. The organic layer was dried (Na2S04) and evaporated and the residue was purified by flash column chromatography (eluting with petroleum ether:EtOAc 100:0 to 5 4:1) to give 157 (36 mg, 47%).
An ice-cold solution of KN03 (117 mg, 1.16 mmol) in 98% H2S04 (1 mL) was added dropwise to a solution of 157 (440 mg, 0.97 mmol) in 98% H2S04 (40 mL) at -12 °C. After 15 mill the mixture was poured into ice water and extracted with cold EtOAc. The organic layer was washed with cold 10 water, cold brine, and then dried (Na2S04) and evaporated. The residue was purified by flash column chromatography (petroleum ether:EtOAc 100:0 to 7:3) to give l-(bromomethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3H-benzo[^]indole-7-sulfonyl chloride (158) as a white powder (287 mg, 59%). A portion was recrystallised from CH?C1,/z'-Pr20 as colourless crystals: mp 217-219 °C; 'H NMR (CDClj) 5 9.35 (s, 1 H), 9.29 (dJ = 1.7 Hz, 1 H), 8.23 (ddj = 9.0, 1.9 Hz, 1 H), 8.11 (dj = 15 9.1 Hz), 4.72 (br dJ = 11.5 Hz, 1 H), 4.61 (ddj = 11.5, 8.9 Hz, 1 H), 4.43 (tt, /= 8.6, 3.0 Hz, 1 H), 3.81 (ddj =11.0, 3.2 Hz, 1 H), 3.58 (ddj =11.0, 7.8 Hz, 1 H). Anal. (C15H9BrClF3N2OsS) Calc: C, 35.91; H, 1.81; N, 5.58. Found: C, 36.00; H, 1.77; N, 5.27. A second product isolated from the column was 1 -(bromomethyl) -9-nitro-3 - (trifluoroacetyl)-1,2-dihydro-3H-benzo [<?]indole-7-sulfonyl chloride (159) (10 mg, 2%). A portion was recrystallised from CH2C12//-Pr20 as colourless crystals: 20 mp 238-243 °C; !H NMR (CDC13) 5 8.91 (dj= 9.1 Hz, 1 H), 8.81 (dj = 1.9 Hz, 1 H), 8.36 (d,/ = 2.0 Hz, 1 H), 8.26 (dj = 9.1 Hz, 1 H), 4.58-4.45 (m, 2 H), 4.18-4.11 (m, 1 H), 3.47 (ddj= 8.1, 3.2 EIz, 1 H), 3.18 (ddj =11.0, 7.3 Hz, 1 H). Anal. (C15HgBrCIF,N205S'0.2*'-Pr20) Calc: C, 37.27; H, 2.28; N, 5.37. Found: C, 37.10; H, 2.11; N, 5.18.
The reaction was repeated on a larger scale with 157 (870 mg, 1.91 mmol) and the crude product 25 purified by crystallisation (CH2Cl2/i-Pr20) to give 158 in higher yield (724 mg, 75%) and free of 159.
A solution of ethanolamine (18 mg, 0.30 mmol) in CH2C12 (0.5 mL) was added dropwise to a solution of 158 (50 mg, 0.10 mmol) in CH2C12(10 mL) at 0 °C. After 15 min Cs2C03 (67 mg, 0.20 mmol), MeOH (3 mL), and water (2 drops) were added. After a further 15 min ice was added and 30 the mixture was poured into a mixture of cold water and cold EtOAc. The organic layer was separated and washed with cold water and cold brine. The organic layer was dried (Na2S04) and evaporated, and the residue was recrystallised from Et0Ac/z'-Pr20 to give 1-(bromomethyl)-IV-(2-hydroxyethyl)-5-nitro-l,2-dihydro-3H-benzo[tf]indole-7-sulfonamide (160) as red crystals (43 mg, 100%): mp 144-147 °C; 'H NMR [(CD3)2SO] § 8.59 (dj = 1.7 Hz, 1 H), 8.02 (dJ = 8.9 Hz, 1 H), 35 7.79 (ddj = 8.9,1.8 Hz, 1 H), 7.69 (tj = 5.9 Hz, 1 H), 6.73 (s, 1 H), 4.63 (tj = 5.6 Hz, 1 H), 4.33- 70 Received at IPONZ on 14 December 2010 4.25 (m, 1 H), 3.89 (tdj = 10.4,1.9 Hz, 1 H), 3.82 (ddj = 10.2, 3.4 Hz, 1 H), 3.74-3.66 (m, 2 H), 3.43-3.30 (m, 3 H), 2.81 (q J = 6.1 Hz, 2 H). Anal. (ClsH1(iBrN3OsS0.1?'-Pr20) Calc: C, 42.54; FI, 3.98; N, 9.54. Found: C, 42.73; H, 3.92; N, 9.30.
A solution of tert b uty I ch lorodi mcthylsilane (21 mg, 0.14 mmol) and DIPEA (30 mg, 0.23 mmol) in DMF (0.5 mL) was added dropwise to a solution of 160 (50 mg, 0.12 mmol) in DMF (2 mL) at 0 °C and the mixture was allowed to warm to room temperature over 1 h. A further portion of tert-butylchlorodimethylsilane (42 mg, 0.28 mmol) and DIPEA (30 mg, 0.23 mmol) were added and after 15 min the mixture was poured into a mixture of cold water and cold EtOAc. The organic layer 10 was separated and washed with cold water and cold brine. The organic layer was dried (Na2S04) and evaporated, and the residue was triturated with petroleum ether and EtzO to give 1-(bromomethyl)-N- [2-(fe^-butyldimethylsilyloxy)ethyl]-5-nitro-1,2-dihydro- 3H-benzo|V|indole-7-sulfonamide (161) as a red powder (60 mg, 95%): mp 69-72 °C (dec.); *H NMR [(CD3)2SO] 5 8.58 (s, 1 EI), 8.02 (d J = 9.0 Hz, 1 H), 7.80 (ddj = 8.9, 1.6 Hz, 1 H), 7.78-7.72 (m, 2 H), 6.74 (s, 1 H), 4.34-4.24 (m, 1 H), 3.88 15 (tj = 10.4 Hz, 1 H), 3.81 (ddj = 10.3, 3.4 Hz, 1 H), 3.74-3.65 (m, 2 H), 3.52 (tJ = 6.3 Hz, 2 H), 2.85 (qj = 6.0 Hz, 2 H), 0.79 (s, 9 H), -0.05 (s, 6 H). Anal. (C21H30BrN305SSi0.3H20) Calc: C, 45.87; EI, 5.61; N, 7.64. Found: C, 45.55; H, 5.28; N, 8.04.
A solution of DIPEA (155 mg, 1.20 mmol) and amine 114 (300 mg, 1.20 mmol) in CH2C12 (1 mL) 20 was added dropwise to a solution of 158 (500 mg, 1.00 mmol) in CH2C12 (10 mL) at -50 °C. After 30 min Cs2C03 (650 mg, 2.0 mmol), MeOH (5 mL), and water (2 mL) were added. The mixture was stirred for 15 min at -10 °C and then poured into a mixture of cold water and cold EtOAc. The organic layer was separated and washed with cold water and cold brine. The organic layer was dried (Na2S04) and filtered through a plug of silica gel (eluting with EtOAc:petroleum ether: 9:1), and the 25 filtrates were evaporated to give di(^-butyl) 2-[l-(bromomethyl)-5-nitro-l,2-dihydro-3H- ben/,o[(?|indole 7 sulfonamide]ethyl phosphate (162) suitable for the following reactions (500 mg, 81%). A portion was recrystallised from CH2C12//-Pr20 as red crystals: mp 76 °C (dec.); 'H NMR [(CD3)2SO] 5 8.59 (dj = 1.6 Hz, 1 H), 8.04 (dJ = 8.9 Hz, 1 H), 7.94 (tJ = 5.6 Hz, 1 FI), 7.80 (ddj = 9.0, 1.8 Hz, 1 H), 7.75 (s, 1 H), 6.75 (s, 1 H), 4.33-4.25 (m, 1 H), 3.89 (tdj = 10.5, 1.9 Hz, 1 H), 30 3.84-3.76 (m, 3 H), 3.74-3.65 (m, 2 H), 3.01 (q J = 5.6 Hz, 2 H), 1.35 (s, 18 H). Anal. (C23H33BrN3OsPS) Calc: C, 44.38; H, 5.34; N, 6.75. Found: C, 44.48; H, 5.20; N, 6.65.
A solution of EDCI.HCl (9.0 g, 58.1 mmol) in CH2C12 (20 mL) was washed with a solution of aqueous 40% K2C03. The organic layer was treated with pyridinium.HBr (7.5 g, 46.5 mmol). Slow 71 Received at IPONZ on 14 December 2010 addition of Et20 to the resulting solution caused a precipitate to form. The solid was filtered off, washed with EtjO, and dried to give EDCI.HBr (8.63 g, 79%) as a white powder.
Biorad AG 1-X4 resin (chloride form, 45 g) was converted to the bromide form by washing with a solution of NaBr (45 g) in water (450 mL). The resin was then washed successively with water (450 5 mL) and MeOH (450 mL). A solution of acid hydrochloride 122 (1.5 g) in MeOH (20 mL) was passed through the resin and the eluate was evaporated to give acid hydrobromide 163 as a cream powder (1.33 g, 78%).
EDCI.HBr (230 mg, 0.98 mmol), anhydrous TsOH (6 mg, 0.033 mmol), and acid 163 (121 mg, 0.33 mmol) were added to a solution of indoline 160 (70 mg, 0.16 mmol) in CH2C12 (15 mL) and DMA (2 10 mL). After 30 min further portions of EDCI.HBr (230 mg, 0.98 mmol) and TsOH (20 mg, 0.12 mmol) were added. After 1.5 h a further portion of acid 163 (60 mg, 0.16 mmol) was added. After a further 5 h the mixture was poured into a mixture of cold water and cold EtOAc. The organic layer was washed with cold water, cold brine, and then dried (Na2S04) and evaporated. The residue was dissolved in a mixed solvent of CH-CL. and MeOH and the solution was slowly concentrated to 15 induce precipitation. The solid was filtered off and washed with MeOH to give 8 as a yellow powder (65 mg, 57%): mp 210-216 °C (dec.); 3H NMR [(CD3)2SO] 5 11.74 (s, 1 H), 9.29 (s, 1 H), 8.85 (d J = 1.5 Hz, 1 H), 8.43 (dJ = 8.9 Hz, 1 H), 8.02 (ddj = 8.9,1.6 Hz, 1 H), 7.91 (tj = 5.9 Hz, 1 H), 7.42 (d J = 8.9 Hz, 1 H), 7.22-7.15 (m, 2 H), 6.96 (dd J = 8.9, 2.3 Hz, 1 EI), 4.97 (t J = 10.5 Hz, 1 H), 4.73-4.62 (m, 3 H), 4.13 (t J = 5.8 Hz, 2 H), 4.08-3.98 (m, 2 H), 3.63-3.56 (m, 4 H), 3.39 (q J = 6.1 20 Hz, 2 H), 2.88 (q J — 6.1 Hz, 2 H), 2.73 (t J — 5.7 EIz, 2 EI), 4 Hs not observed. Anal.
(C30El32BrN5O8S-y3H2O) Calc: C, 50.64; H, 4.67; N, 9.84. Found: C, 50.47; H, 4.60; N, 9.67.
Example 22. 2- [l-(Bromomethyl)-3-[5-(2-morpholinoethoxy)-l/£indole-2-carbonyI]-5-mtro-l,2-dihydro-3//-benzo[e]iiidole-7-sulfona.mido] ethyl dihydrogen phosphate trifluoroacetate 25 (23) (Scheme L). EDCI.HBr (prepared as described in Example 21, 3.74 g, 15.8 mmol), anhydrous TsOH (109 mg, 0.63 mmol) and acid 163 (1.30 g, 3.48 mmol) were added to a solution of indoline 162 (1.97 g, 3.17 mmol) in CH2C12 (55 mL) and DMSO (5 mL). After 1 h further portions of EDCI.HBr (3.0 g, 12.7 mmol), anhydrous TsOH (200 mg, 1.16 mmol) and indoline 162 (600 mg, 1.61 mmol) were added. After a further 3 h the mixture was poured into a mixture of cold water and 30 cold EtOAc. The organic layer was washed with cold water, cold brine and then dried (Na2S04) and concentrated to a small volume to precipitate the crude product. The solid was filtered off and triturated with acetone to give di-/cr/-butyl 2-{l-(bromomethyl)-3-[5-(2-morpholinoethoxy)-lH-mdole-2-carbonyl]-5-nitro-1,2-dihy dro-3H-benzoje\indole 7 sulfcmamido}ethyl phosphate (188) as a yellow powder (1.08 g, 38%): mp 228-233 °C (dec.); lU NMR [(CD3)2SO] 5 11.73 (s, 1 H), 9.30 (s, 1 35 H), 8.87 (dJ = 1.7 Hz, 1 H), 8.45 (dj = 8.9 Hz, 1 H), 8.18 (tJ = 5.9 Hz, 1 H), 8.02 (ddJ = 8.9, 72 Received at IPONZ on 14 December 2010 1.7 Hz, 1 H), 7.42 (dJ = 8.9 Hz, 1 H), 7.23-7.17 (m, 2 H), 6.96 (ddj = 8.9, 2.4 Hz, 1 H), 4.97 (tJ = 10.4 Hz, 1 H), 4.73-4.63 (m, 2 H), 4.13 (tj = 5.7 Hz, 2 H), 4.08-3.99 (m, 2 H), 3.83 (qJ = 6.0 Hz, 2 H), 3.60 (t J = 4.5 Hz, 4 H), 3.07 (q J = 5.8 Hz, 2 H), 2.74 (t J = 5.4 Hz, 2 H), 1.36 (s, 18 H), 4 Hs not observed. Anal. (C^H^Br^O^PS'V^O) Calc: C, 50.50; H, 5.58; N, 7.75. Found: C, 5 50.68; H, 5.57; N, 7-82.
A solution of 164 (140 mg, 0.16 mmol) and TFA (178 mg, 1.56 mmol) in CH2C12 (5 mL) was stirred at room temperature for 20 h. Removal of solvents gave 23 as a yellow powder (146 mg, 100%): mp 206-210 °C (dec.); 'H NMR [(CD3)2SO] 8 11.78 (s, 1 H), 9.28 (s, 1 H), 8.87 (dj = 1.5 Hz, 1 H), 8.42 10 (dJ = 8.9 Hz, 1 H), 8.19 (br s, 1 H), 8.01 (ddj = 8.9, 1.5 Hz, 1 H), 7.43 (dJ = 8.9 Hz, 1 H), 7.24 (dJ = 2.0 Hz, 1 H), 7.21 (dj = 1.4 Hz, 1 H), 6.99 (ddJ = 8.9, 2.3 Hz, 1 H), 4.97 (tJ = 10.5 Hz, 1 H), 4.72-4.63 (m, 2 H), 4.27 (t J = 5.9 Hz, 2 H), 4.08-3.97 (m, 2 H), 3.82 (q J = 6.1 Hz, 2 FI), 3.87-3.70 (m, 2 H), 3.09-2.95 (m, 4 H), 9 Hs not observed, obscured by water peak. Anal. (C32H34BrF3N5013PS) Calc: C, 42.87; H, 3.82; N, 7.81. Found: C, 43.21; H, 4.11; N, 7.73.
Example 23. (£)-l-(Bfomomethyl)-A^(2-hydfoxyethyl)-3-{3-[4-(2- morpholinoethoxy)phenyl] acryloyl}-5-nitro-l,2-dihydro-3i7-benzo[e]indole-7-sulfonamide (9) (Scheme M). Biorad AG 1-X4 resin (chloride form, 45 g) was converted to the bromide form by washing with a solution of NaBr (45 g) in water (450 mL). The resin was then washed 20 successively with water (450 mL) and MeOH (450 mL). A solution of acid hydrochloride 129 (1.0 g) in MeOFI (20 mL) was passed through the resin and the eluate was evaporated to give acid hydrobromide 165 as a cream powder (1.0 g, 88%).
EDCI.HBr (prepared as described in Example 21, 329 mg, 1.40 mmol), anhydrous TsOH (8 mg, 0.047 mmol) and acid 165 (166 mg, 0.47 mmol) were added to a solution of indoline 160 (100 mg, 25 0.23 mmol) in CH2C12 (8 mL) and DMA (1 mL). After 2 h further portions of acid 165 (166 mg, 0.47 mmol), EDCI.HBr (329 mg, 1.40 mmol) and anhydrous TsOH (8 mg, 0.047 mmol) were added. After a further 2 h the mixture was poured into a mixture of cold water and cold EtOAc. The organic layer was washed with cold water, cold brine and then dried (Na2S04) and evaporated. The residue was dissolved in a mixed solvent of CH2C12 and MeOH and the solution was slowly 30 concentrated to induce precipitation. The solid was filtered off and washed with MeOH to give 9 as a yellow powder (91 mg, 57%): mp 185-190 °C (dec.); *H NMR [(CD3)2SO] 8 9.34 (s, 1 H), 8.83 (s, 1 H), 8.35 (d, J = 8.8 Hz, 1 H), 8.01 (d, J ~ 8.9 Hz, 1 H), 7.90 (tj = 5.8 Hz, 1 H), 7.79 (d, J = 8.7 Hz, 2 H), 7.73 (dj = 15.3 Hz, 1 H), 7.09 (dJ = 15.4 Hz, 1 H), 7.03 (dJ = 8.7 Hz, 2 H), 4.73-4.63 (m, 3 H), 4.59-4.51 (m, 1 H), 4.16 (t J = 5.7 Hz, 3 H), 4.05-3.92 (m, 3 H), 3.63-3.55 (m, 4 H), 3.54-3.47 73 Received at IPONZ on 14 December 2010 (m, 3 PI), 2.86 (q J = 6.0 Hz, 3 H), 2.72 (tj = 5.7 Hz, 2 H). Anal. (C30H33BrN4O8S'CH3OH) Calc: C, 51.60; H, 5.17; N, 7.76. Found: C, 51.89; H, 5.07; N, 7.60.
Example 24. (ii)-2-[l-(Bromomethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-5 l,2-dihydro-3.£/-beiizo[<?]indole-7-sulfonamido]ethyl dihydrogen phosphate trifluoroacetate (24) (Scheme M). EDCI.HBr (prepared as described in Example 21, 820 mg, 3.47 mmol), anhydrous TsOH (20 mg, 0.12 mmol) and acid 165 (410 mg, 1.16 mmol) were added to a solution of indoline 162 (360 mg, 0.58 mmol) in CH2C12 (15 mL) and DMA (1 mL). After 3 h the mixture was poured into a mixture of cold water and cold EtOAc. The organic layer was washed with cold water, 10 cold brine and then dried (Na2S04) and evaporated. The residue was dissolved in a mixed solvent of CH2C12 and MeOH and the solution was slowly concentrated to induce precipitation. The solid was filtered off and washed with MeOH to give (£)-di-^nf-butyl 2-[1 -(bromomethyl)-3- {3- [4 - (2 morpholinoethoxy)phenyl]acryloy}-5-nitro-1,2-dihydro-3H-benzo[s'Jindolc 7 sulfonamido)ethyl phosphate (166) as a yellow powder (250 mg, 50%): mp 177-182 °C (dec.); H NMR [(CD3)2SO] 8 15 9.35 (s, 1 H), 8.86 (dJ = 1.7 Hz, 1 H), 8.39 (dj = 8.9 Hz, 1 H), 8.16 (tj = 5.9 Hz, 1 H), 8.00 (ddj = 8.9, 1.7 Hz, 1 H), 7.79 (dJ = 8.8 Hz, 2 H), 7.73 (dj = 15.3 Hz, 1 H), 7.10 (dj= 15.3 Hz, 1 H), 7.03 (d J = 8.8 Hz, 2 H), 4.71-4.63 (m, 2 H), 4.61-4.53 (m, 1 FI), 4.17 (t J = 5.7 Hz, 2 H), 4.04-3.93 (m, 2 H), 3.82 (q, j = 6.0 Hz, 2 H), 3.59 (t, J = 4.6 FIz, 4 H), 3.06 (q J = 5.8 Hz, 2 H), 2.72 (t, J = 5.7 Hz, 2 H), 1.35 (s, 18 H), 4 Hs not observed. Anal. (C38H5(lBrN4011PS) Calc: C, 51.76; FI, 5.72; N, 20 6.35. Found: C, 51.80; H, 5.91; N, 6.21.
A solution of 166 (250 mg, 0.28 mmol) and TFA (320 mg, 2.84 mmol) in CH2C12 (12 mL) was stirred at room temperature for 20 h. Removal of solvents and trituration of the residue with Et,0 gave 24 as a yellow powder (250 mg, 100%): mp 178-182 °C (dec.); 'H NMR [(CD3)2SO] 8 9.35 (s, 1 H), 8.85 25 (s, 1 H), 8.38 (dj = 8.8 Hz, 1 H), 8.18-8.08 (m, 1 H), 8.01 (dj = 8.4 Hz, 1 H), 7.86 (dj = 8.4 Hz, 2 H), 7.76 (dj = 15.2 Hz, 1 FI), 7.14 (dj = 15.2 Hz, 1 H), 7.11 (dj = 8.4 Hz, 2 H), 4.73-4.63 (m, 2 H), 4.60-4.52 (m, 1 H), 4.46-4.36 (m, 2 H), 4.07-3.94 (m, 2 FI), 3.89-3.74 (m, 4 H), 3.09-2.98 (m, 2 H), 11 Hs not observed, obscured by water peak. Anal. (C32H35BrF3N4013PS'H20) Calc: C, 42.63; H, 4.14; N, 6.21. Found: C, 42.93; H, 4.25; N, 6.02. 74 Received at IPONZ on 14 December 2010 In Vivo Activity of Compounds of Formula III.
The ability of compounds of Formula III to eliminate colony forming cells of human tumor xenografts grown in nude mice (CDl-Foxnlnu) was determined using an excision assay. Tumors were grown subcutaneously in the flank by inoculating cells grown in tissue culture (107 cells in 100 5 p,L). When tumors reached treatment size (500-700 mm3) mice were randomised to treatment groups (3 mice per group for radiation or drug alone, 5 mice per group for compound plus radiation). Compounds were dissolved in phosphate buffered saline containing 2-4 equivalents of NaHCOj and given as single intravenous doses alone or 5 min after whole body irradiation (' 'Co source). Eighteen hours after treatment tumors were excised, weighed, minced, dissociated 10 enzymatically, and plated to determine clonogenicity as described (Hicks et al.,/. Nail. Cancer Inst., 2006, 98, 111 8-1128). Clonogens per gram of tumor tissue were calculated relative to controls: for compound alone relative to no treatment, and for compound plus radiation relative to radiation alone.
Log extra cell kill in excision assay3 Compound alone Compound +15 Gy radiationb No SiHa H460 H1299 HCT116 SiHa H460 H1299 HCT116 22Rvl RT 0.63 0.24 0.85 0.92 1.36 0.27 1.82 0.97 - R2 0.26 0.00 - - 0.86 0.07 - - - R3 0.31 0.03 - - 0.57 -0.03 - - - R4 0.34 - - - 0.55 - - - - R5 0.08 0.12 - - 0.32 0.05 - - - R6 0.17 -0.13 - - 0.30 0.09 - - - 1.16 0.13 1.06 - 2.41 0.76 >2.89 >2.60 1.18 16 1.26 0.61 1.39 1.79 2.26 0.79 2.24 >2.40 - 17 1.05 0.48 - - 2.58 1.12 - - - 18 -0.01 0.09 - - 1.54 0.44 - - - 19 1.49 0.25 - - 1.47 0.12 - - _ 0.36 - - - 1.72 0.54 - - - 21 0.79 - - - 2.19 - - - - 22 0.92 - - - 2.20 - - - - 23 1.32 0.12 - - 2.80 0.39 - - - 24 0.45 - - - 1.56 - - - - 75 Received at IPONZ on 14 December 2010 aThe compounds were administered at the following doses: 42 (j,mol/kg for SiHa, 56 (.imol/kg for HCT116, and 75 jamol/kg for H460 and H1299, with the exception of R3 dosed at 24 (J.mol/kg in SiHa and H460, R5 dosed at 13 (imol/kg in H460,17 dosed at 56 |J.mol/kg in H460, and 21, 22, and 24 dosed at 30 |^mol/kg in SiHa. bRadiation alone provides following log cell kill: 2.04 (SiHa), 1.43 (H460), 1.26 (H1299), 2.21 (HCT116).
Structures of reference compounds (Rl-4 from WO 2006/043839) shown below.
O (HOfePO^fl! o Os/~ NMe2 O ii (HOfePOv CI iWSB o no2 R2 N IVIes O {HOfcPO^flf o o^ NEt2 OMe N T OMe H OMe O {HO^RX ■Ms o CI .o. o o II (HO)2PO.
NO, CI . Vn ,o o 1 o N02 R5 R6 This assay provides evidence of hypoxia selective cell lulling in vivo. A whole body radiation dose of 15 Gy is sufficient to sterilize well oxygenated tumor cells, and provides 1-2 logs of cell kill in these xenograft models. Additional cell kill when radiation is combined with the compounds of Formula 15 III (15-24) shows that these compounds are eliminating hypoxic tumor cells. The compounds of Formula III are significantly more active in this regard than the reference compounds of WO 2006/043839 (Rl-4). The compounds of Formula III are also significantly more active than two reference compounds R5 and R6, which both incorporate a morpholine in the sidechain, but not one which is attached to an indole (Formula la) or cinnamate (Formula lb) sidechain. The 20 difference between the various reference compounds and the examples of this invention is illustrated in Figures 3a and 3b, respectively, for SiHa and H460 xenografts. The difference is also 76 Received at IPONZ on 14 December 2010 notable for the HI299 and HCT116 xenografts in Table 4. Table 4 also shows evidence of single agent activity. This may be as a consequence of a bystander effect where the compounds of Formula III are activated in hypoxic regions of tumors and the reduced metabolites diffuse to and kill surrounding better oxygenated tumor cells. The compounds of Formula III are generally superior to 5 the reference compounds of WO 2006/043839 in single agent activity.
The superior in vivo activity of the compounds of Formula III is unexpected since the structures of the reference compounds Rl-4 of WO 2006/043839 are very similar to the structures of the examples of Formula III (15-24 using the numbering in Table 3). The superior in vivo activity of the 10 compounds of Formula III is particularly unexpected since reference compounds which include a morpholine in the sidechain (R5 and R6), but not one which is attached to an indole or cinnamate sidechain, show very little activity in vivo.

Claims (38)

Received at IPONZ on 14 December 2010 Although this invention has been described with reference to certain embodiments and examples, those persons skilled in the art will appreciate that the specific description is illustrative only and that variations may be made without departing from the scope of the invention. Received at IPONZ on 14 December 2010 CLAIMS
1. A compound of Formula I: no2 5 wherein Y is selected from CI or Br, and wherein X is selected from S02NR12 or CONR:2, where X is located at either position 7 or 8, and where each R1 independently represents H or a C,_4alkyl, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb 10 R3 c / \ t / \ or2 (lb) (Ic) (Id) wherein R2 represents a lower C2_4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) substituent, and R3 represents FI, C^ alkyl, C^alkoxy, C,_4 alkynyl or C,_4 alkynyloxy, 15 and pharmaceutically acceptable salts thereof.
2. A compound of Formula I: 7 n02 wherein Y is selected from CI or Br, 20 and wherein X is selected from S02NR12 or CONR1,, where X is located at either position 7 or ! and where each R1 independently represents H or a Chalky 1, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb 79 Received at IPONZ on 14 December 2010 (la) (lb) (ic) (id) wherein R2 represents a lower C2_4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) substituent, and R3 represents H or CV4 alkyl, 5 and pharmaceutically acceptable salts thereof.
3. A compound according to claim 2, wherein R3 represents H.
4. A compound according to claim 3, wherein Y is CI. 10
5. A compound according to claim 3, wherein Y is Br.
6. A compound according to claim 4 or 5, wherein, X is 7-S02NH(CH2)20H. 15
7. A compound according to claim 4 or 5, wherein X is 7-CONFI(CF[2)2OH.
8. A compound according to any one of claims 2 to 7, wherein Z is selected from the group consisting of:
9. A compound of Formula I according to claim 2, selected from the group consisting of: 1-(Chloromethyl)-JV-(2-hy droxy ethyl)-3-[5-(2-morp h olinoeth oxyr) -1 H-indole-2-carbonyl]-5-nitr o-1,2 -dihydro benzoj<?|indolc 7 sulfonamide; 25 l-(Chloromethyl)-IV-(2-hydroxyethyl)-3-[5-(3-morpholinopropoxy)-lH-indole-2-carbonyl]-5-nitro-1,2-dihydro-3H-benzo [ejindole- 7-sulfonamide; 80 Received at IPONZ on 14 December 2010 (E) -1 - (Chloromethyl)-iV- (2 -hydroxy ethyl)-3- { 3 - [4-(2-morphoIinoethoxy) phenyl] acryloyl} - 5-nitro-1,2-dihy dro-3H-b enzo [tfjindole -7 - sulfonamide; l-(Chloromethyl)-AT-(2-hydroxyethyl)-3-{4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-l,2-dihydro-3_H-benzo [^indoles-sulfonamide; 5 l-(Chloroinethyl)-A/-(2-hydi'oxyethyI)-3-[5-(2-morpholinoethoxy)-lH-indole-2-carbonyl]-5-nitto-l,2- dihydro-3FEbenzo [tf]indole-7-carb oxamide; (E) -1 - (chlotomethyl) -N-(2-hydroxyethyl)-3- { 3-[4-(2-morpholinoethoxy) p henyl] acryloyl} - 5-nitro-1,2-dihydro-3H-benzo|y]indole-7-carboxamide; (i) ■-1 - (Chloromethyl) -N- (2-hydroxyethyl) -3 - [5-(2-morpholinoethoxy)-1 H-indole-2- carbonyl] 5-nitr o-10 1,2-dihy dro-3H-benzo\e\ indole-7-sulfonamide; 1 -(Bromomethyl)-iV-(2-hydro xyethyl)-3- [5- (2-morpholinoethoxy) -1H i n do 1 e 2 ■ c a r b o n y 11 - 5 - nitro -1,2-dihydro 3f7 benzo|?|indole 7 sulfonamide; and (E)-l -(Bromomethyl)-N-(2-hydroxyethyl)-3- {3-[4-(2-morpholinoethoxy)phenyl]acryloyl} -5-nitro-1,2-dihy dro-3H-benzo [<?] indole-7-sulfonamide; 15 and pharmaceutically acceptable salts thereof
10. A compound of Formula II: NH2 wherein Y is selected from CI or Br, 20 and wherein X is selected from S02NR12 or CONR '2, where X is located at either position 7 or 8, and where each R1 independently represents H or a C, alkyl, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb wherein R2 represents a lower C2_4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) substituent, and R3 represents H, a Ct_4 alkyl, C1_4alkoxy, C,_4 alkynyl or C,^ alkynyloxy, and pharmaceutically acceptable salts thereof. 81 Received at IPONZ on 14 December 2010
11, A compound of Formula II, (id wherein Y is selected from CI or Br, and wherein X is selected from SOjNR1, or CONR1,, where X is located at either position 7 or 8, and where each R1 independently represents H or a C^alkyl, optionally substituted with one or more hydroxyl groups, and wherein Z is a sidechain selected from either structure la or lb 10 0a) Cb) (Ic) (Id) wherein R2 represents a lower C2 4 alkyl bearing a morpholine (Ic) or N-methylpiperazine (Id) substituent, and R1 represents Ft or C[ A alkyl, and pharmaceutically acceptable salts thereof. 15
12. A compound according to claim 11, wherein R3 represents H.
13. A compound according to claim 12, wherein Y is CI. 20
14. A compound according to claim 12, wherein Y is Br.
15. A compound according to claim 13 or 14, wherein X is 7-S02NFl(CFI2)20H.
16. A compound according to claim 13 or 14, wherein X is 7-CONH(CH2)2OH. 25
17. A compound according to any one of claims 11 to 16, wherein Z is selected from the group consisting of: 82 Received at IPONZ on 14 December 2010
18. A compound of Formula II according to claim 11, selected from the group consisting of: 5 Amino 1 -(chloromethyl)-N-(2-hydroxyethyl)-3-[5-(2-morpholinoethoxy)-1 H-indole-2 -carbonyl]-1,2 di hydro 3 R l.)cn7o[ejindolc-7-su1forianu.de; 5-Amino-1 - (chloromethyl) -N- (2-hydroxyethyl) -3- [5-(3-morpholinopropoxy) -1 H-indole-2-carb onyl] -1,2-dihy dro-3H-benz o [e]indole-7-sulfonamide; (j5)-5-Amino-l-(chloromethyl)-iV-(2-hydroxy ethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-1,2-diliydro-3ii-b enzo [^indole-? -sulfonaniide; 5-Amino-l - (chloromethyl)-iV-(2-hydroxyethyl)-3- [5- (2-morpholinoethoxy)-1 H-indole-2-carbonyl] -1,2-dihydro-.3 H-benzo [.?j indole 7 carboxamidc; and (i)-5-Amino-1 - (chloromethyl)-N-(2-hydroxy ethyl) -3- [5-(2-morpholinoethoxy)-1 H-indole-2-carbonyl] -1,2-dihy dro-3H-benzo [tf] indole-7-sulfonamide; and pharmaceutically acceptable salts thereof.
19. A compound of Formula Ilia, lllb, IIIc or Illd: Formula 1 0 11 Formula 1 —0 r r4 i i or —op(oh)2 or if n" Formula II Formula II [o HJ (lEIa) (lllb) Formula I or Ho Formula II o- ,r£ ( Formula I or Formula II "TRq 9 *op(oh)2 (|llc) (Illd) wherein Formula I and the substituents thereof are as defined in claim 1 and Formula II and the substituents thereof are as defined in claim 10, and any compound of Formula I or Formula II, bearing a free hydroxyl group is functionalised with a phosphate (Formula Ilia) or an amino acid or short polypeptide chain (Formula lllb) or a monosaccharide (Formula IIIc), or wherein any 83 Received at IPONZ on 14 December 2010 compound of Formula I or Formula II bearing a free hydroxyl group or a secondary or tertiary amine is functionalised with a phosphonooxymethyl substituent (Formula Hid), wherein R4 is selected from any substituent found in the naturally occuring amino acids, m is selected from 1, 2, 3, or 4, R5 represents any combination of hydroxy, hydroxymethyl, carboxylic acid or other 5 substituents found in naturally occuring monosaccharides, p represents 0 or 1, T represents O or N, and when T represents N it represents the secondary amine of the indole la or the tertiary amine of the morpholine Ic or either of the tertiary amines of the piperazine Id, wherein R represents the remainder of the structure of Formula I or Formula II compatible with these definitions, and q represents 0 when T represents O and q represents 1 or 2 when T represents N, and when q 10 represents 2 the charge of the quaternary ammonium group of Formula Illd is balanced by an appropriate anion, and pharmaceutically acceptable salts thereof.
20. A compound of Formula Ilia, lllb, IIIc of Illd: Formula 1 o II Formula 1 -o r r4 i i or —OP(OH)2 or Formula II Formula 11 11 H O m (ilia) (lllb) TRq 9 ^OP(OH)2 15 (Hie) (Illd) wherein Formula I and the substituents thereof are as defined in claim 2 and Formula II and the substituents thereof are as defined in claim 11, and any compound of Formula I or Formula II, as defined in claim 2 or 11, bearing a free hydroxyl group is functionalised with a phosphate (Formula Ilia) or an amino acid or short polypeptide chain (Formula lllb) or a monosaccharide (Formula 20 IHc), or wherein any compound of Formula I or Formula II bearing a free hydroxyl group or a secondary or tertiary amine is functionalised with a phosphonooxymethyl substituent (Formula Hid), wherein R4 is selected from any substituent found in the naturally occuring amino acids, m is selected from 1, 2, 3, or 4, R5 represents any combination of hydroxy, hydroxymethyl, carboxylic acid or other substituents found in naturally occuring monosaccharides, p represents 0 or 1, T 25 represents O or N, and when T represents N it represents the secondary amine of the indole la or the tertiary amine of the morpholine Ic or either of the tertiary amines of the piperazme Id, wherein Pnrmi i Id I Wl I 1 IU IC4 or Formula I I I I UIIIIUICI I or Formula II 84 Received at IPONZ on 14 December 2010 R represents the remainder of the structure of Formula I or Formula II compatible with these definitions, and q represents 0 when T represents O and q represents 1 or 2 when T represents N, and when q represents 2 the charge of the quaternary ammonium group of Formula Illd is balanced by an appropriate anion, 5 and pharmaceutically acceptable salts thereof.
21. A compound according to claim 20 wherein R3 represents H.
22. A compound according to claim 21 wherein Y is CI. 10
23. A compound according to claim 21 wherein Y is Br.
24. A compound according to any one of claims 21 to 23 wherein X is 7 S()2NI I (CI QXMI, and is functionalized with a phosphate, amino acid, short polypeptide chain, monosaccharide or 15 phosphonooxymethyl substituent as defined in claim 20.
25. A compound according to any one of claims 21 to 23 wherein X is 7-CONH(CFI2)2OH, and is functionalized with a phosphate, amino acid, short polypeptide chain, monosaccharide or phosphonooxymethyl substituent as defined in claim 20. 20
26. A compound according to any one of claims 20 to 23 which is a compound of Formula Ilia.
27. A compound according to claim 26 wherein X is functionalized with a phosphate group to 25 form the group 7-S02NFI(CFI2)20P0(0FE)2 .
28. A compound according to claim 26 wherein X is functionalized with a phosphate group to form the group 7-CONFI(CFl2)2OPO(OF[)2 30 29.
A compound according to any one of claims 20 to 28, wherein Z is selected from the group consisting of: Received at IPONZ on 14 December 2010 r? n h C D
30. A compound of Formula Ilia according to claim 20, selected from the group consisting of: 2- [1 - (Chloromethyl) -3 - [5- (2-morpholinoethoxy) -1 H-indole-2-carbonyl] -5 -nitro-1,2-dihydro-3.H-5 benzo[?]indole-7-sulfonamido] ethyl dihydrogen phosphate; 2-[l-(Chloromethyl)-3-[5-(3-morpholinopropoxy)-lH-indole-2-carbonyl]-5-nitro-l,2-dihydro-3H-benzo[e]mdole-7-sulfonamido] ethyl dihydrogen phosphate; f/Ej-2-(l-(chlorome(iiyl)-3- {3-[4-(2-morpholinoclhoxy) phenyl] acryloyl} 5 nitro-1,2-dihydro-3ii-benzo[e]indole-7-sulfonamido)ethyl dihydrogen phosphate; 10 2-[l-(Chloromethyl)-3- {4-[2-(4-methylpiperazin-l-yl)ethoxy]benzoyl}-5-nitro-l ,2-dihydro-3fi-benzo[tf]indole-7-sulfonamido]ethyl dihydrogen phosphate; 2-{l~(Chloromethyl)-3-[5-(2-morpholinoethoxy)-lH-indole-2-carbonyl]-5-nitro-l,2-dihydro-3H-benzo[i?]indole-7-carboxamido} ethyl dihydrogen phosphate; (£)-2-[l -(Chloromethyl)-3- {3-[4-(2-morpholinoethoxy)phenyl]acryloyl} -5-nitro-l ,2-dihydro-3H- (J)-2- [1 -(Chloromethyl)-3- [5-(2-morpholinoethoxy) -1H-indole-2-carbonyl] -5-nitro-1,2-dihydro-3H-benzo[<?]indole-7-sulfonamido] ethyl dihydrogen phosphate; (S, jE)-2-[1 -(Chloromethyl)-3- {3-[4-(2-morpholinoethoxy)phenyl] acryloyl} -5-nitro-l ,2-dihydro-3H-benzo [^indole-7-sulfonamido] ethyl dihydrogen phosphate; 20 2-[l-(Bromomethyl)-3-[5-(2-morpholinoethoxy)-l.H-indole-2-carbonyl]-5-nitro-l,2-dihycko-3J:f-benzo[<?]indole-7-sulfonamido] ethyl dihydrogen phosphate; and (E)-2-[l-(Bromomethyl)-3-{3-[4-(2-morpholinoethoxy)phenyl]acryloyl}-5-nitro-l ,2-dihydro-3H-benzo[<?]indole-7-sulfonamido] ethyl dihydrogen phosphate. 25
31. A pharmaceutical composition comprising a compound of Formula I, as defined in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carder.
32. A pharmaceutical composition comprising a compound of Formula II, as defined in claim 10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. Received at IPONZ on 14 December 2010
33. A pharmaceutical composition comprising a compound of Formula Ilia, lllb, IIIc or Hid, as defined in claim 19, or a pharmaccuticallv acceptable salt thereof, and a pharmaceutically acceptable carrier.
34. The use of a compound of Formula I, II or III as defined in claim 1, 10 or 19 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the production of an anti-cancer effect in a warm-blooded animal such as a human.
35. The use of a compound of Formula I as defined in claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a cancer in a warm-blooded animal such as a human.
36. The use of a compound of Formula II as defined in claim 10 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a cancer in a warmblooded animal such as a human.
37. The use of a compound of Formula Ilia, Illh, IIIc or Illd as defined in claim 19 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a cancer in a warm-blooded animal such as a human.
38. A use according to any one of claims 34 to 37, wherein the cancer is selected from the group consisting of cervical cancer, non-small cell lung carcinoma, large cell lung carcinoma, colon cancer and prostate cancer.
NZ571028A 2008-09-03 2008-09-03 Nitrobenzindole compounds and their use in cancer treatment NZ571028A (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
NZ571028A NZ571028A (en) 2008-09-03 2008-09-03 Nitrobenzindole compounds and their use in cancer treatment
PCT/NZ2009/000184 WO2010027280A1 (en) 2008-09-03 2009-09-03 Nitrobenzindoles and their use in cancer therapy

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
NZ571028A NZ571028A (en) 2008-09-03 2008-09-03 Nitrobenzindole compounds and their use in cancer treatment

Publications (1)

Publication Number Publication Date
NZ571028A true NZ571028A (en) 2011-01-28

Family

ID=41797304

Family Applications (1)

Application Number Title Priority Date Filing Date
NZ571028A NZ571028A (en) 2008-09-03 2008-09-03 Nitrobenzindole compounds and their use in cancer treatment

Country Status (2)

Country Link
NZ (1) NZ571028A (en)
WO (1) WO2010027280A1 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9901567B2 (en) 2007-08-01 2018-02-27 Syntarga B.V. Substituted CC-1065 analogs and their conjugates
BRPI0921687A8 (en) 2008-11-03 2022-11-08 Syntarga Bv COMPOUND, CONJUGATE, USE OF A COMPOUND, PHARMACEUTICAL COMPOSITION, PROCESS FOR PREPARING A PHARMACEUTICAL COMPOSITION, METHOD FOR TREATING A MAMMAL IN NEED THEREOF, AND, METHOD FOR TREATING OR PREVENTING A TUMOR IN A MAMMAL.
PL3056203T3 (en) * 2010-04-21 2018-06-29 Syntarga B.V. Conjugates of cc-1065 analogs and bifunctional linkers
WO2014062856A1 (en) 2012-10-16 2014-04-24 Halozyme, Inc. Hypoxia and hyaluronan and markers thereof for diagnosis and monitoring of diseases and conditions and related methods
PT3092010T (en) 2014-01-10 2018-09-28 Synthon Biopharmaceuticals Bv Method for purifying cys-linked antibody-drug conjugates
KR102344354B1 (en) 2014-01-10 2021-12-28 비온디스 비.브이. Duocarmycin adcs showing improved in vivo antitumor activity
KR20220045075A (en) 2014-01-10 2022-04-12 비온디스 비.브이. Duocarmycin adcs for use in treatment of endometrial cancer

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003097635A1 (en) * 2002-05-17 2003-11-27 Auckland Uniservices Limited Processes for preparing 3-substituted 1-(chloromethyl)-1,2-dihydro-3h-[ring fused indol-5-yl(amine-derived)] compounds and analogues thereof, and to products obtained therefrom
NZ536107A (en) * 2004-10-22 2007-06-29 Auckland Uniservices Ltd Nitrobenzindoles and their use in cancer therapy

Also Published As

Publication number Publication date
WO2010027280A1 (en) 2010-03-11

Similar Documents

Publication Publication Date Title
NZ571028A (en) Nitrobenzindole compounds and their use in cancer treatment
DK2822953T5 (en) Macrocyclic derivatives for the treatment of proliferative diseases
US8993614B2 (en) Substituted pyrrolidine-2-carboxamides
US9597341B2 (en) Phosphate compounds, a process for their preparation and pharmaceutical compositions containing them
KR101596526B1 (en) Parp inhibitor compounds, compositions and methods of use
CA2169179C (en) 5-arylindole derivatives
AU617304B2 (en) Novel cc-1065 analogs
AU2005296389B2 (en) Nitrobenzindoles and their use in cancer therapy
CA2924705A1 (en) Quinazoline derivative and preparation method therefor
CA2858799A1 (en) Tricyclic boron compounds for antimicrobial therapy
EP2854800A1 (en) Substituted pyrrolidine-2-carboxamides
CA3014395A1 (en) Carboxamide derivatives useful as rsk inhibitors
US8183247B2 (en) Boron compounds useful in BNCT
US20120065170A1 (en) Antimicrobial Cyclocarbonyl Heterocyclic Compounds For Treatment Of Bacterial Infections
WO2018038988A2 (en) Compounds, compositions, methods for treating diseases, and methods for preparing compounds
TW202313606A (en) Substituted phenyl-1h-pyrrolo[2,3-c]pyridine derivatives
SK288085B6 (en) Indole derivatives, medicament comprising said derivatives, use of said derivatives and process for their preparation
KR20110082094A (en) 6-cycloamino-3-(1h-pyrrolo[2,3-b]pyridin-4-yl)imidazo[1,2-b]pyridazine derivatives, preparation thereof and therapeutic use thereof
KR20170095243A (en) Heterocyclyl linked imidazopyridazine derivatives as pi3kbeta inhibitors
US20070191372A1 (en) Dna-targeted benzotriazine 1,4-dioxides and their use in cancer therapy
CA2903276A1 (en) Muscarinic agonists
KR100729980B1 (en) Bis-n,n&#39;-bis-2-haloethylaminophosphoramidates as antitumor agents
CZ2019667A3 (en) Purine nucleoside phosphorylase inhibitors, preparing and using them to treat T-cell acute lymphoblastic leukemia and lymphomas
CN115109055B (en) Difunctional compound for EGFR degradation and application thereof
JP2023514364A (en) Macrocyclic Indole Derivatives as Inhibitors of MCL-1

Legal Events

Date Code Title Description
ASS Change of ownership

Owner name: AUCKLAND UNISERVICES LIMITED, NZ

Free format text: OLD OWNER(S): AUCKLAND UNISERVICES LIMITED; MOANA TERCEL; FREDERIK BASTIAAN PRUIJN; RALPH JAMES STEVENSON; SHANGJIN YANG; HO HUAT LEE; JEAN-JACQUES YOUTE TENDOUNG

PSEA Patent sealed
LAPS Patent lapsed