WO2008039087A2 - Indoline derivatives and uses thereof - Google Patents

Indoline derivatives and uses thereof Download PDF

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Publication number
WO2008039087A2
WO2008039087A2 PCT/NZ2007/000285 NZ2007000285W WO2008039087A2 WO 2008039087 A2 WO2008039087 A2 WO 2008039087A2 NZ 2007000285 W NZ2007000285 W NZ 2007000285W WO 2008039087 A2 WO2008039087 A2 WO 2008039087A2
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WIPO (PCT)
Prior art keywords
methyl
benz
dihydro
amino
oxo
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PCT/NZ2007/000285
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French (fr)
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WO2008039087A8 (en
WO2008039087A3 (en
Inventor
Stephen Mark Stribbling
Kathleen Grace Mountjoy
Moana Tercel
William Robert Wilson
William Alexander Denny
Ralph James Stevenson
Guo-Liang Lu
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Auckland Uniservices Limited
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Priority claimed from AU2006905415A external-priority patent/AU2006905415A0/en
Application filed by Auckland Uniservices Limited filed Critical Auckland Uniservices Limited
Publication of WO2008039087A2 publication Critical patent/WO2008039087A2/en
Publication of WO2008039087A3 publication Critical patent/WO2008039087A3/en
Publication of WO2008039087A8 publication Critical patent/WO2008039087A8/en

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    • 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
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links

Definitions

  • the present invention relates to compounds that are indoline derivatives.
  • the invention also relates to intermediates used in the synthesis of such indoline derivatives and to pharmaceutical compositions containing such indoline derivatives.
  • the invention is also concerned with methods of utilising these compounds in prophylactic or therapeutic treatment of obesity, weight gain, metabolic disorders resulting in obesity or weight gain and associated conditions such as hyperglycaemia, hypertriglyceridemia and diabetes.
  • the invention will be hereinafter described with reference to such application.
  • the object of the present invention is to overcome or ameliorate at least some of the disadvantages of the prior art therapies or to provide a useful alternative.
  • the present invention provides novel indoline derivatives, found to be effective in reducing body weight in mammals and thus useful in treatment of conditions resulting in undesirable or excessive body weight gain, metabolic disorders resulting in these conditions and the more serious complications such as diabetes (in particular Type II diabetes), cardiovascular disease and the like.
  • n 2 or 3 and when n is 3 the N bears a positive charge which is balanced by a counterion of negative charge
  • R 1 is selected from NH 2 , OH 5 NHR, NR 2 where R is lower alkyl
  • R 2 is selected from CH 2 OH, CH 2 Cl, CH 2 Br, CH 2 I, CH 3 , CH 2 OSO 2 R where R is lower alkyl R 3 is selected from one of the following
  • m is selected from 1 to 8 p is selected from 1 to 3
  • X is selected from N or CH the point of attachment is selected from the sites marked 3, 4, 5, or 6
  • R 4 is selected from R (lower alkyl), CH 2 Ph, CH 2 (substituted Ph) or CH 2 (heterocycle or substituted heterocycle); and pharmaceutically acceptable salts thereof.
  • the preferred compounds can be selected from the following, provided herein merely as non-limiting examples: 5-Amino-l-(cMoromethyl)-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]-l,2- dihydro-3H-benz[e]indole (SN28125);
  • a pharmaceutical composition comprising a compound of the first aspect and a pharmaceutically acceptable carrier.
  • a method of reducing body weight of a subject comprising the administration to a subject of a compound according to the first aspect.
  • a method of prophylactic or therapeutic treatment of a metabolic or an eating disorder resulting in undesirable body weight gain comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
  • a method of prophylactic or therapeutic treatment of obesity comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
  • a method of treating hyperglycaemia comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
  • a method of treating hypertriglyceridemia comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
  • an eighth aspect of the present invention there is provided a method of treating Type II diabetes comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
  • a ninth aspect of the present invention there is provided use of a compound according to the first aspect in the manufacture of a medicament for treatment of a condition selected from the group consisting of: excess body weight, metabolic disorders resulting in excess body weight, obesity, high blood glucose and triglycerides and diabetes.
  • a process for the preparation of a compound according to the first aspect includes but is not limited to the following key steps: 1.
  • the quatemisation is generally carried out at room temperature in the dark over several days in the presence of a slight excess of a suitable alkyl halide. ⁇ In cases where the quaternary salt remains soluble the reaction may be accelerated by the use of inorganic bases (e.g. the use OfK 2 CO 3 in the formation of 50 and 51). 3.
  • inorganic bases e.g. the use OfK 2 CO 3 in the formation of 50 and 51. 3.
  • the formation of a quaternary salt by direct quaternisation of a compound containing a potentially reactive aniline or phenol functional group. This is illustrated with the synthesis of 10 in Scheme 1 , 37 in Scheme 4, and SN27829 in
  • Figure IA Effect of a single injection of 4.2 ⁇ mol/kg SN28127 on body weight, food and water intake on 4 different strains of mice (the data are mean ⁇ SEM).
  • a single injection of SN28127 induces long term weight loss in four different strains of mice.
  • Four different strains of male mice C3HeN, a/a, A w /a, and NZG) aged 50-70 days were housed individually and each animal received a single ip injection of either DMSO (control) or SN28127 (4.2 ⁇ mol/kg) in DMSO (drug). Body weight, food and water intake were measured daily for 160 days post injection.
  • mice Four different strains of male mice (C3HeN, a/a, A ⁇ / ⁇ /a, and NZG) aged 50-70 days were housed individually and each animal received a single ip injection of either DMSO (control) or SN28127 (4.2 ⁇ mol/kg) in DMSO (drug). The mice were culled at -160 days post injection and bled from the retro-orbital sinus. Plasma was prepared and used for biochemical measurements and insulin and leptin concentrations were measured using commercial Eliza assay kits.
  • FIG. 1 Effect of a single injection of SN29220 on body weight, food and water intake on C3HeN mice aged ⁇ 50 days (the data are mean for 4 mice).
  • a single injection of SN29220 induces weight loss in C3HeN mice in a dose dependent manner.
  • Male C3HeN mice aged 50-70 days were housed in groups of 4 per cage and each animal received a single ip injection of either DMSO (control) or SN29220 (0.47 ⁇ mol/kg, 1.4 ⁇ mol/kg, or 4.2 ⁇ mol/kg) in DMSO.
  • Body weight food and water intake were monitored and the mice were culled at 60 days post injection.
  • FIG. 3 Effect of 0.42 ⁇ mol/kg SN29728 on body weight of C3HeN mice.
  • the mice were aged approximately 50 days at the start of the experiment and received two drug doses, one at day 0 and a repeat injection at day 39 (the data are mean for 4 mice).
  • SN29728 causes reversible weight loss in C3HeN mice.
  • Male C3HeN mice aged 50-70 days were housed in groups of 4 per cage and each animal received a single ip injection of either DMSO (control) or SN29728 (0.42 ⁇ mol/kg) in DMSO.
  • the mice received a second injection 39 days after the first injection.
  • the data shows that there was a transient reduction in body weight following the first SN29728 injection and a further decrease in body weight following the second injection.
  • Figure 4 Reversal of high fat diet induced obesity in male C57BL/6J mice with
  • FIG. 5 Reversal of insulin resistance in male C57BL/6J mice fed a high fat diet with SN29220.
  • Male C57BL/6J mice were housed in groups of 4 per cage and fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492 ⁇ ) diet from day 21 (weaning) and treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 ⁇ mol/kg body weight) on day 80.
  • mice on low fat diet and treated with DMSO On day 42 post injection, two mice on low fat diet and treated with DMSO, three mice on high fat diet and treated with DMSO, and three mice on high fat diet and treated with 1.4 ⁇ mol/kg SN29220 were fasted overnight. The following morning glucose tolerance tests were performed on these mice. Blood glucose was monitored using a glucometer (ACCU-CHEK Advantage, Roche) on a drop of blood collected from the tail tip of each mouse. Immediately after this each mouse received a single ip injection of glucose (2g/kg body weight). At 30 minute intervals over 180 minutes following the glucose injection, blood glucose was monitored using a glucometer on a drop of blood collected from the tail tip of each mouse.
  • a glucometer ACCU-CHEK Advantage, Roche
  • the present invention is directed to compounds of general Formula (I):
  • n 2 or 3 and when n is 3 the N bears a positive charge which is balanced by a counterion of negative charge
  • R 1 is selected from NH 2 , OH, NHR, NR 2 where R is lower alkyl
  • R 2 is selected from CH 2 OH 5 CH 2 Cl, CH 2 Br, CH 2 I, CH 3 , CH 2 OSO 2 R where R is lower alkyl
  • R 3 is selected from one of the following
  • m is selected from 1 to 8 p is selected from 1 to 3
  • X is selected from N or CH the point of attachment is selected from the sites marked 3, 4, 5, or 6
  • R 4 is selected from R (lower alkyl), CH 2 Ph, CH 2 (substituted Ph) or CH 2 (heterocycle or substituted heterocycle); and pharmaceutically acceptable salts thereof. Examples of suitable compounds according to the invention are provided in Table 1, below.
  • Tetrahydrocyclopropa[c]benz[e]indol-4-one CBI An Enhanced and Simplified Analog of the CC- 1065 and Duocarmycin Alkylation Submits. J. Org. Chem. (1995) 60(5), 1271-1275. This document describes the synthesis of 41, and, in references cited therein, the deprotection of 41 and formation of amides from this. These publications will enable the skilled person to produce compounds within the scope of Formula (I) by utilising common general knowledge and/or the synthesis methods disclosed.
  • the salts of the compounds of this invention refer to nontoxic "pharmaceutically acceptable salts.”
  • Other salts may, however, be useful in the preparation of compounds according to this invention or of their pharmaceutically acceptable salts.
  • Suitable pharmaceutically acceptable salts of the compounds include the counterion of the quaternary ammonium species, which may be formed during the quaternisation reaction, or by ion exchange chromatography using a resin bearing the conjugate base of a pharmaceutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid.
  • Suitable pharmaceutically acceptable salts of the compounds include acid addition salts which may, for example, be formed by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid.
  • a pharmaceutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid.
  • the compounds of the invention may be prepared as zwitterionic species, or as suitable pharmaceutically acceptable salts of the acidic moiety, these salts may include alkali metal salts, e.g. sodium or potassium salts; alkaline earth metal salts, e.g. calcium or magnesium salts; and salts formed with suitable organic ligands, e.g. quaternary ammonium salts.
  • alkali metal salts e.g. sodium or potassium salts
  • alkaline earth metal salts e.g. calcium or magnesium salts
  • suitable organic ligands e.g. quaternary ammonium salts.
  • representative pharmaceutically acceptable salts include the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydro chloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate,
  • the present invention includes within its scope prodrugs of the compounds of this invention.
  • prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the required compound.
  • the term “administering” shall encompass the treatment of the various disorders described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient.
  • Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", Ed. H. Bundgaard, Elsevier, 1985, incorporated in its entirety herein by reference.
  • the compounds according to this invention may accordingly exist as enantiomers. Where the compounds possess two or more chiral centres, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are encompassed within the scope of the present invention. Furthermore, some of the crystalline forms for the compounds may exist as polymorphs and as such are intended to be included in the present invention. In addition, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are also intended to be encompassed within the scope of this invention. The invention also encompasses active metabolites of a compound of the invention. Pharmaceutical Compositions
  • the present invention also includes within its scope pharmaceutical compositions comprising, as an active ingredient, at least one compound of the invention or a pharmaceutically acceptable salt or prodrug or hydrate thereof together with a pharmaceutically acceptable carrier or diluent.
  • compositions containing a compound of the invention of the present invention may be prepared by conventional techniques, e.g. as described in "Remington: The Science and Practise of Pharmacy", 19 th Edn., 1995.
  • the compositions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications.
  • compositions include a compound of the invention or a pharmaceutically acceptable basic addition salt or prodrug or hydrate thereof, associated with a pharmaceutically acceptable excipient which may be a carrier or a diluent or be diluted by a carrier, or enclosed within a carrier which can be in the form of a capsule, sachet, paper or other container.
  • a pharmaceutically acceptable excipient which may be a carrier or a diluent or be diluted by a carrier, or enclosed within a carrier which can be in the form of a capsule, sachet, paper or other container.
  • the active compound will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet, paper, or other container.
  • the carrier When the carrier serves as a diluent, it may be solid, semi- solid, or liquid material, which acts as a vehicle, excipient, or medium for the active compound.
  • the active compound can be adsorbed on a granular solid container for example in a sachet.
  • suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatine, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclo dextrin, amylose, magnesium stearate, talc, gelatine, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycei ⁇ des and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone.
  • the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax.
  • the formulations may also include wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavouring agents.
  • the formulations of the invention may be formulated so as to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art.
  • compositions can be sterilised and mixed, if desired, with auxiliary agents, emulsifiers, salt for influencing osmotic pressure, buffers and/or colouring substances and the like, which do not deleteriously react with the active compounds.
  • the route of administration may be any route, which effectively transports the active compound of the invention to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal or parenteral e.g. rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment, the oral route being preferred.
  • the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
  • the preparation may contain a compound of the invention dissolved or suspended in a liquid carrier, in particular an aqueous carrier, for aerosol application.
  • a liquid carrier in particular an aqueous carrier
  • the carrier may contain additives such as solubilizing agents, e.g. propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclo dextrin, or preservatives such as parabenes.
  • injectable solutions or suspensions preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor oil.
  • Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application.
  • Preferable carriers for tablets, dragees, or capsules include lactose, corn starch, and/or potato starch.
  • a syrup or elixir can be used in cases where a sweetened vehicle can be employed.
  • the compounds of Formula (I) may be administered to a mammal, especially a human in need of such treatment, prevention, elimination, alleviation or amelioration of the various disorders such as metabolic disorders, excessive or undesirable weight gain, obesity Type II diabetes, hyperglycaemia, hypertriglyceridemia, disorders of food intake and related conditions.
  • the compounds of Formula (I) are contemplated to be useful for the prevention or treatment of obesity.
  • the compounds of Formula (I) may be useful for blood glucose lowering, prevention of hyperglycemia, delaying the progression of impaired glucose tolerance to Type II diabetes, delaying the progression of non- insulin requiring Type II diabetes to insulin requiring Type II diabetes, or appetite regulation.
  • the mammal to be treated with a compound of Formula (I) is preferably a human, but may also be a domesticated animal, e.g. household pet or farmyard animal, and non-domesticated animal such as wildlife.
  • the compounds of the invention are effective over a wide dosage range. From the experimental studies described herein the representative compounds typically showed activity from about 300 ⁇ g/kg to about 3000 ⁇ g/kg. For example, in the treatment of adult humans, dosages from about 20mg to about 250 mg per day may be used and may be divided into multiple doses. The dosage need not be daily and the requirement for administration would be gauged by the response of an individual. In choosing a regimen for patients it may frequently be necessary to begin with a higher dosage and when the condition is under control to reduce the dosage. The exact dosage will depend upon the mode of administration, on the therapy desired, form in which administered, the subject to be treated and the body weight of the subject to be treated, and the preference and experience of the physician or veterinarian in charge.
  • the compounds of the present invention are dispensed in unit dosage form comprising from about 20 mg to about 50 mg of active ingredient together with a pharmaceutically acceptable carrier per unit.
  • the invention furthermore relates to the use of a compound according to the present invention for the preparation of a medicament for use in the treatment of excessive or undesirable weight gain, obesity and associated conditions in a regimen which additionally comprises treatment with another anti-obesity or anti-weight gain agent.
  • the compounds of the present invention may be administered in conjunction with a known anti-diabetic agent.
  • the expression "antidiabetic agent” includes compounds for the treatment and/or prophylaxis of insulin resistance and diseases wherein insulin resistance is the pathophysiological mechanism.
  • the compounds may also be administered in conjunction with a hypoglycaemic agent, another anti-obesity agent or an antilipidemic agent.
  • Example 5 Synthesis of iV-4- ⁇ (£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H r - benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-N ⁇ V-dimethyl-A r -[(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN28127) Dioxane (3 mL) saturated with HBr gas was added to a suspension of 5 (144 mg,
  • Example 7 Synthesis of A7-4- ⁇ (jE)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3i ⁇ - benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl- ⁇ yV-dimethyl-iV-[(l-methyl-4-nitro-5- imidazolyl)methyl] ammonium bromide hydrobromide (SN28126) Dioxane (1 mL) saturated with HBr gas was added to a suspension of 6 (54 mg,
  • Example 8 Synthesis of iV-4-((jE)-3- ⁇ 5-[bis(to ⁇ -butoxycarbonyl)]amino-l- (chloromethyI)-l,2-dihydro-3Jjr-benz[e]indol-3-yl ⁇ -3-oxo-l-propenyl)benzyl-iV ⁇ V- dimethyl-JV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide (7)
  • Example 9 Synthesis of 7V-4- ⁇ (£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H- benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl- ⁇ VV-dimethyl-iV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) Dioxane (2 mL) saturated with HBr gas was added to a solution of 7 (40 mg, 0.046 mmol) in dioxane (1 mL) and the mixture was stirred in the dark at 20 °C for 15 h.
  • the mixed salt 110 mg was dissolved in MeOH (2 mL) and purified by ion exchange chromatography (Biorad AG- 1X4 resin in Cl " form), eluting with MeOH.
  • Oxalyl chloride (187 mg, 1.54 mmol) and then DMF (1 drop) were added to a suspension of 16 (76 mg, 0.171 mmol) in dry CH 3 CN (3 mL). The mixture was stirred for 2 h and then evaporated under reduced pressure. The amine prepared above was dissolved in THF (3 mL) with Et 3 N (58 mg, 0.57 mmol) and the resulting mixture was added to the acid chloride in THF (2 mL). The mixture was stirred for 2 h., the solvents were removed under reduced pressure, and EtOAc and H 2 O were added to the residue.
  • Example 14 Synthesis of iV-4- ⁇ (£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3iy- benz[ ⁇ ]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-iV-ben2yl- ⁇ VV-dimethyl] ammonium bromide (10)
  • the aldehyde 21 (3.08 g, 19.9 mmol) was dissolved in MeOH (70 mL) and treated with NaBH 4 (0.75 g, 19.8 mmol). After 30 min at 20 °C aqueous NaCl was added and the MeOH was evaporated. The aqueous residue was extracted with EtOAc (x5) and the extracts were dried (Na 2 SO 4 ) and evaporated, to give 5-(Hydroxymethyl)-l- methyl-4-nitroimidazole 22 as a cream solid (2.68 g, 86%). A sample was recrystallised from EtOAc, m.p.
  • Example 24 Synthesis of 2-(bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole (25)
  • a solution OfBr 2 (373 mg, 2.33 mmol) in dry CH 3 CN (5 mL) was added dropwise to a cooled solution of Ph 3 P (527 mg, 2.33 mmol) in dry CH 3 CN (9 mL) at 0 °C.
  • Ph 3 P 527 mg, 2.33 mmol
  • a solution of 24 [Chou et ah, Tetrahedron Lett. (2001) 42, 1309-1311] (400 mg, 2.12 mmol) in dry CH 3 CN (3 mL) was added at 20 °C.
  • Example 26 Synthesis of 5-[bis(to ⁇ -butoxycarbonyl)]amino-l- ⁇ [(tert- butoxycarbonyl)-oxy]methyl ⁇ -3-(trifluoroacetyl)-l,2-dihydro-3J9 r -benz[e]indole (29)
  • a solution of 27 200 mg, 0.588 mmol) in THF (15 niL) and MeOH (3 mL) was hydrogenated over PtO 2 -XH 2 O (Acros, Pt content 79-84%, 30 mg) at 40 psi for 1 h.
  • the aniline 28 (180 mg, 0.581 mmol) was dissolved in dioxane (10 mL) with di- tert-butyl dicarbonate (1.01 g, 4.65 mmol) and DMAP (5 mg) and the solution was stirred at reflux under nitrogen for 15 h. More di-tert-butyl dicarbonate (506 mg, 2.32 mmol) was added and the solution stirred at reflux for a further 5 h. The solvent was evaporated and the residue purified by column chromatography (petroleum ether:EtOAc 9:1).
  • Oxalyl chloride (187 mg, 1.48 mmol) and then DMF (1 drop) were added to a suspension of E-4-(dimethylaminomethyl)cinnamic acid hydrochloride (89 mg, 0.369 mmol) in dry CH 3 CN (8 mL). The mixture was stirred for 3 h and then the solvents were removed.
  • Example 29 Synthesis of N-(4- ⁇ (jE)-3-[5-amino-l-(hydroxymethyl)-l,2-dihydro- 3£T-benz[e]indoI-3-yl]-3-oxo-l-propenyl ⁇ benzyl)-iV,iV-dimethyl-N-[(l-methyl-5- nitro-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29293)
  • Example 30 Synthesis of iV-4-((£)-3- ⁇ 5-[bis(to ⁇ -butoxycarbonyl)]amino-l-[(te ⁇ - butoxycarbonyI)oxy]methyl-l,2-dihydro-3 J H r -benz[e]indoI-3-yl ⁇ -3-oxo-l- propenyl)benzyl- ⁇ yV-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2-pyrrolyl)- methyl] ammonium bromide (32)
  • Example 31 Synthesis of ⁇ -(4- ⁇ (£)-3-[5-ammo-l-(hydroxymethyl)-l,2-dihydro- 3i ⁇ -benz[ ⁇ ]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyI)-iVyV-dimethyl-iV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29221) Dioxane (3 mL) saturated with HBr gas was added to a solution of 32 (98 mg, 0.103 mmol) in dioxane (2 mL) and the mixture was stirred in the dark at 20 0 C for 15 h.
  • Example 33 Synthesis of 5-allylamino-3-[(JE)-4- (dimethylammomethyl)cinnamoyl]-l-methyl-l,2-dihydro-3jH-benz[e]indole (35)
  • TFA 15 mL
  • 34 (1.40 g, 3.20 mmol) at 0 0 C and the solution was stirred at 20 0 C for 2 h.
  • the TFA was removed under reduced pressure and the residue was partitioned between CH 2 Cl 2 and aqueous NaHCO 3 .
  • the CH 2 Cl 2 layer was washed with brine, dried (Na 2 SO 4 ), and evaporated to give the deprotected amine (760 mg, 100%) which was used directly in the following steps.
  • Example 35 Synthesis of ⁇ - ⁇ ( ⁇ -S-IS-amino-l-methyl-l ⁇ dihydro-Sl ⁇ - benz[e]indol-3-yl] -3-oxo-l-propenyl ⁇ benzyl-iV-benzyl-iV, ⁇ f-dimethyl] ammonium . bromide (37)
  • Example 37 Synthesis of iV-4-((E)-3- ⁇ 5-[bis(te/ ⁇ i 1 -butoxycarbonyl)]amino-l-methyl- l ? 2-dihydro-3 J fiT-benz[e]indol-3-yl ⁇ -3-oxo-l-propenyl)benzyl- ⁇ VV-dimetb.yl-iV-[(l- methyI-5-nitro-2-pyrrolyl)methyl] ammonium bromide (39)
  • Example 38 Synthesis of N-4- ⁇ (£)-3-[5-amino-l-methyl-l,2-dihydro-3iy- benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-i ⁇ yV-dimethyI-iV-[(l-methyl-5-nitro-2- pyrrolyl)methyl] -ammonium chloride hydrochloride (SN29873)
  • Example 39 Synthesis of iV-4-((E)-3- ⁇ 5-[bis(te/"/-butoxycarbonyl)]amino-l-inethyl- l,2-dihydro-3H r -benz[e]indol-3-yl ⁇ -3-oxo-l-propenyl)benzyl]-N ! ⁇ V-dimethyl-A'-[(l- methyl-5 ⁇ methylsuIfonyl-2-pyrrolyl)methyl] ammonium bromide (40)
  • Example 42 Synthesis of iV ⁇ - ⁇ -S-fl-Cchloromethy ⁇ -S-hydrosy-ljl-dihydro-SJ ⁇ - benz[e] -indol-3-yl] -3-oxo-l-propenyl ⁇ benzy 1-JV ⁇ -dimethyl-iV- [(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide (SN27829)
  • 6-(Dimethylamino)hexanoyl chloride hydrochloride [Olomucld, Bull. Soc. CUm. Fr. (1963), 2067-2074] (203 mg, 0.953 mmol) in THF (5 mL) was added slowly to a solution of the amine prepared above in CH 2 Cl 2 (5 mL). The mixture was stirred for 30 min, then diluted with H 2 O and extracted with CH 2 Cl 2 . The CH 2 Cl 2 extract was washed with brine, dried (Na 2 SO 4 ), and evaporated.
  • Example 50 Synthesis of 6-[5-[bis(/ ⁇ ? ⁇ -butoxycarbonyl)amino]-l-(chloromethyl)- l,2-dihydro-3J ⁇ -benzo[ ⁇ indol-3-yl]-iV ⁇ V-dimethyI-iV- ⁇ [l-methyI-5-(methy ⁇ suIfonyl)- l/2 r -pyrrol-2-yl]methyl ⁇ -6-oxo-l-heptane ammonium bromide (51)
  • Example 54 Synthesis of iV-7- ⁇ 5-[bis(tert-butoxycarbonyl)amino]-l- (chIoiOmethyl)-l,2-dihydro-3fi r -benz[-?]indol-3-yl ⁇ -7-oxo-l-heptanyl-i ⁇ yV, ⁇ '- trimethyl ammonium iodide (53)
  • Example 55 Synthesis of iV-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3 J H r - benz[e]indol-3-yl]-7-oxo-l-heptanyl-i ⁇ yV,iV-triinethylaminoniuin chloride hydrochloride (SN31652)
  • Example 56 Synthesis of iV-7- ⁇ 5-[bis(tert-butoxycarbonyl)amino]-l- (chloromethyl)-l,2-dihydiO-3Jg r -benz[e]indol-3-yl ⁇ -7-oxo-l-heptanyl- ⁇ v /V-dimethyl- iV-4-nitrobenzylammonium bromide (54)
  • Example 57 Synthesis of iV-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3i ⁇ - benz[e]indol-3-yl]-7-oxo-l-heptanyl- ⁇ yV-dimethyl-iV-4-nitrobenzylammonium chloride hydrochloride (SN31653) To a solution of 54 (43 mg, 0.054 mmol) in CH 2 Cl 2 (1 mL) was added HCl in dioxane (4M, 5 mL). The mixture was stirred for 8 h and the supernatant was decanted off.
  • Example 58 Synthesis of A ⁇ -V-IS-lbis ⁇ ert-butoxycarbony ⁇ amino]-!- (chloromethyl)-l,2-dihydro-3jH r -benz[e]indol-3-yl ⁇ -7-oxo-l-heptanyl-A r -4- cyanobenzyl-iNyV-dimethylammonium bromide (55)
  • a solution of 49 (40 mg, 0.068 mmol) and 4-cyanobenzyl bromide (15 mg, 0.075 mmol) in dry MeCN (4 mL) was stirred at room temperature for 18.5 h.
  • Example 59 Synthesis of A ⁇ 7-[5-amino-l-(chIoromethyl)-l,2-dihydro-3i?- benz[e]indoI-3-yl]-7-oxo-l-heptanyl-iV-4-cyanobenzyl-7V ⁇ V-dimethylammonium chloride hydrochloride (SN31666)
  • the bisBOC compound 43 (62 mg , 0.13 mmol) was treated with HCl in dioxane (4M, 3 mL) and the mixture stirred at room temperature for 3 h. The solvent was evaporated and the off-white solid obtained (the amine hydrochloride) was used directly in a subsequent step.
  • the amine hydrochloride prepared above was suspended in CH 2 Cl 2 (5 mL) and the suspension was cooled in an ice bath. Diisopropylethyl amine (0.30 mL) was added, giving a clear solution. A suspension of the acyl chloride hydrochloride prepared above in CH 2 Cl 2 (5 mL) was added slowly, giving a yellow solution, which was stirred at ice bath temperature for 30 min and at room temperature for 1.5 h.
  • Example 61 Alternative synthesis of 5-amino-l-(chloromethyl)-3-[(£)-4- (dimethylaminometliy ⁇ -cinnamoyll-ljl-dihydro-SH-benztelindole CSNlSllS)
  • a solution of 56 (26 mg, 0.057 mmol), PhSO 2 Na (24 mg, 0.14 mmol), and camphorsulfonic acid (40 mg, 0.17 mmol) in dry CH 2 Cl 2 (10 mL) was purged by bubbling with a stream of nitrogen gas.
  • Pd(PPh 3 ) 4 (5 mg, 0.0043 mmol) was added and the mixture was stirred for 1.5 h.
  • Example 62 Alternative synthesis of 5-[bis(fert-butoxycarbonyl)]amino-l- (chloromethyl)-3- [(£)-4-(dimethylaminomethyl) cinnamoyl] -1 ,2-dihy dro-3i ⁇ - benz[e]indole (4)
  • Example 63 Synthesis of iV-4- ⁇ (jE)-3-[5-bis(tert-butoxycarbonyl)amino-l- (chloromethylJ-l ⁇ -dihydro-SjH-benzI ⁇ indol-S-yll-S-oxo-l-propenylJbenzyl-iV- benzyl-AyV-dimethyl] ammonium bromide (57)
  • Example 64 Synthesis of iV-4- ⁇ (£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3i ⁇ - benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-i ⁇ VV-dimethyl-7V-(4- methylsulfonyl)benzylammonium chloride hydrochloride (SN31630)
  • Example 65 General Animal Model Procedures The general procedures to which the animals were subjected are as follows: 1. Application of ear tags under anaesthesia (3% halothane/O 2 ) to provide individual animal identification. Using a stainless steel small animal ear punch one or two holes were punched out close to the edge of either one or both ears of a mouse. This system allows a numbering system to identify individually up to 20 mice in one cage. For the majority of the mice only one hole was punched.
  • mice Weighing of mice by removal from their cages and placing in a box on a top- loading animal balance.
  • mice treated with SN28127 a representative compound of the invention, can lose much more than 15% of their body weight with no deterioration in their health and that this weight change is not a secondary consequence of loss of appetite or difficulty in eating or drinking.
  • Example 66 Effect of ⁇ - ⁇ ( ⁇ -S-tS-amino-l ⁇ chloromethy ⁇ -l ⁇ -dihydro-Sff- benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-iV v /V-diinethyl-A L [(l-methyl-5-nitro-2- pyrrolyl)methy I] ammonium bromide hydrobromide (SN28127) on Four Different Strains of Mice
  • mice Male mice (approx. 40 days) were house individually and had unrestricted access to routine chow and tap water. Mouse body weight, food and water intake were monitored daily Monday through Friday. Each mouse was weighed carefully by removing from its cage, placing it in a cardboard box on top of a balance and then returning it to its cage. Baseline body weight, food and water intake were measure for 7- 10 days prior to injection to determine that the animals were eating and gaining weight. In all four mouse strains, SN28127 caused significant reductions in body weight without decreased food and water intake (see Figure IA). Treatment with SN28127 actually increased food and water intake between 10 and 80 days post drug injection. After this time, food and water intake were very similar to control animals despite the drug treated mice continuing to weigh significantly less than control mice. At 160 days post injection, all 4 strains had normal glucose levels (see Figure IA).
  • mice were identified by ear-tagging under anaesthetic (3% halothane/O 2 ) prior to treatment.
  • Mouse body weight, food and water intake were monitored by weighing 2-3 times per week. Food and water intake were averaged per mouse for each cage. Each mouse was weighed by carefully removing from its cage, placing it in a cardboard box on top of a balance and then returning it to its cage. Baseline body weight, food and water intake were measured for 7-10 days prior to injection to determine that the animals were eating and gaining weight.
  • Example 68 Reversible Effect of ⁇ - ⁇ (J ⁇ -S-IS-amino-l ⁇ chloromethyl)-! ⁇ - dihydro-3i ⁇ -benz[e]indol-3-yl]-3-oxo-l-propenyl ⁇ benzyl-iVyV-dimethyl-iV-[(l- methyl-5-nitro-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN28127) on Weight Loss.
  • mice housed as described in Example 67 were treated with a single injection of the SN28127 analogue SN29728 (0.42 ⁇ mol/kg body weight) on day zero . Their body weight decreased from day 12 through day 30 post injection. After day 30 the body weight increased. Following a second SN29220 injection (0.42 ⁇ mol/kg body weight) at day 39 body weight was again reduced and stabilised at a reduced level for a further 60 days (see Figure 3).
  • Example 69 ⁇ - ⁇ ( ⁇ -S-IS-amino-l ⁇ chloromethyO-l ⁇ -dihydro-SJ ⁇ -benz ⁇ indol-S- yl]-3-oxo-l-propenyl ⁇ benzyl- ⁇ yV-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) reverses high fat diet induced obesity.
  • the body weights of SN29220 treated mice on the high fat diet decreased from day 5 post injection compared to DMSO treated mice on high fat diet. The extent of weight loss was dependent on the dose of SN29220.
  • Example 70 iV- ⁇ f ⁇ -S-lS-amino-l-Cchloromethy ⁇ -l ⁇ -dihydro-SH-benzI ⁇ indol-S- yl]-3-oxo-l-propenyl ⁇ benzyl- ⁇ yV-dimethyl-iV-[(l-methyl-5-methyIsulfonyl-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) reverses high fat diet induced insulin resistance.
  • mice housed as described in Example 67 were fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492i) diet from day 21 (weaning) and treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 ⁇ mol/kg body weight) on day 80.
  • DMSO or SN29220 (0.47 or 1.4 ⁇ mol/kg body weight
  • the area under the curve was greater for the DMSO control mice on the high fat diet compared to the area under the curve for SN29220 treated mice on high fat diet and age-matched control mice on low fat diet indicating that SN29220 completely reverses high fat diet induced insulin resistance.

Abstract

The present invention relates to compounds which are indoline derivatives of formula (I), wherein Rl, R2, R3, R4 and n are as defined in the specification, intermediates used in their synthesis and pharmaceutical compositions containing these compounds. The invention is also concerned with methods utilising these compounds in prophylactic or therapeutic treatment of obesity, weight gain, metabolic disorders resulting in obesity or weight gain and associated conditions such as high blood glucose and triglycerides, and Type II diabetes.

Description

INDOLINE DERIVATIVES AND USES THEREOF
Field of the Invention
The present invention relates to compounds that are indoline derivatives. The invention also relates to intermediates used in the synthesis of such indoline derivatives and to pharmaceutical compositions containing such indoline derivatives. The invention is also concerned with methods of utilising these compounds in prophylactic or therapeutic treatment of obesity, weight gain, metabolic disorders resulting in obesity or weight gain and associated conditions such as hyperglycaemia, hypertriglyceridemia and diabetes. The invention will be hereinafter described with reference to such application.
However, those skilled in the art will readily appreciate that the invention is not limited to such fields of use. Background of the Invention
Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.
Obesity has reached epidemic proportions globally and is now affecting developing as well as developed countries, and is expected to get worse. One hundred and fifty four million people worldwide suffer from obesity, a serious life threatening disease that is a major risk factor for hypertension, stroke, heart disease, diabetes, and cancer. This imposes a heavy burden on the healthcare systems of many industrialised countries, causing up to 10% of total health costs. The global epidemic of obesity results from increased food intake and decreased physical activity. While diet and exercise are recommended as treatments for obesity they are mostly ineffective. Despite the high prevalence of obesity and the many advances in our understanding of how it develops, current therapies have persistently failed to achieve long-term success. Today there are no safe and well-tolerated drugs available to induce substantial long-term weight loss. Thus there is a need for improved or alternative therapies in the treatment of excessive weight gain, obesity and associated conditions such as diabetes, cardiovascular disease and the like.
The object of the present invention is to overcome or ameliorate at least some of the disadvantages of the prior art therapies or to provide a useful alternative. Summary of the Invention
The present invention provides novel indoline derivatives, found to be effective in reducing body weight in mammals and thus useful in treatment of conditions resulting in undesirable or excessive body weight gain, metabolic disorders resulting in these conditions and the more serious complications such as diabetes (in particular Type II diabetes), cardiovascular disease and the like.
According to a first aspect of the present invention there is provided a compound of general Formula (I):
Figure imgf000003_0001
wherein n is 2 or 3 and when n is 3 the N bears a positive charge which is balanced by a counterion of negative charge
R1 is selected from NH2, OH5 NHR, NR2 where R is lower alkyl
R2 is selected from CH2OH, CH2Cl, CH2Br, CH2I, CH3, CH2OSO2R where R is lower alkyl R3 is selected from one of the following
Figure imgf000003_0002
wherein m is selected from 1 to 8 p is selected from 1 to 3
X is selected from N or CH the point of attachment is selected from the sites marked 3, 4, 5, or 6
R4 is selected from R (lower alkyl), CH2Ph, CH2(substituted Ph) or CH2(heterocycle or substituted heterocycle); and pharmaceutically acceptable salts thereof.
The preferred compounds can be selected from the following, provided herein merely as non-limiting examples: 5-Amino-l-(cMoromethyl)-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]-l,2- dihydro-3H-benz[e]indole (SN28125);
N-4-{(E)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo-l-propenyl}benzyl-iV,iV-dimethyl-N-[(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN28127);
N-4-{(E)-3-[5-artiino-l-(chloromethyl)-l;2-dihydro-3H-beiiz[e]indol-3-yl]-3- oxo - 1 -propenyl } benzyl-Λζ,N-dimethyl-N- [(I -methyl-4-nitro -5 - imidazolyl)methyl] ammonium bromide hydrobromide (SN28126);
N-4-{(£)-3-[5-arnmo-l-(chloromethyl)-l,2-dihydro-3H-beiiz[e]indol-3-yl]-3- oxo-l-proρenyl}berizyl-iV,iV-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl]ammonium bromide hydrobromide (SN29220);
N-(4-{(E)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo-l-propenyl}benzyl)-iV,iV,N-trimethylammonium chloride hydrochloride (SN29728); l-(Chloromethyl)-3-{(E)-3-[4-({methyl[(l-methyl-5-nitro-2- pyrrolyl)methyl]amino}methyl)phenyl]-2-piOpenoyl}-l,2-dihydro-3//'-benz[e]indol-5- amine dihydro chloride (SN29532);
N-4-{(^-3-[5-arnino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo-1 -propenyl} benzyl-N-benzyl-iV,iV-dimethyl] ammonium bromide (10);
N-(4-{(E)-3-[5-arnino-l-(hydroxymetliyl)-l,2-dihydro-3H-beiiz[e]indol-3-yl]-3- oxo- 1 -propenyl}benzyl)-ΛζiV-dimethyl-N-[(l -methyl-5-nitro-2- pyrrolyl)methyl]ammonium bromide hydrobromide (SN29293);
N-(4-{(E)-3-[5-amino-l-(hydroxymethyl)-l,2-dihydiO-3H-benz[e]indol-3-yl]-3- oxo-l-propenyl}benzyl)-Λr,iV-dimethyl-iV:-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl]ammonium bromide hydrobromide (SN29221); ΪV-4- {(E)-3-[5-amino-l -methyl- 1 ,2-dihydro-3//-benz[e]indol-3-yl]-3-oxo-l - propenyl}benzyl-N-benzyl-N,N-dimethyl]ammonium bromide (37);
N-4-{(E)-3-[5-arnino-l-methyl-l,2-diliydro-3H-benz[e]indol-3-yl]-3-oxo-l- propenyl}benzyl-Λζ,N-dhO.ethyl-N-[(l-methyl-5-nitro-2-pyrrolyl)methyl]arniiionium chloride hydrochloride (SN29873); N-4-{(^-3-[5-arniiio-l-methyl-l,2-dihydro-3H-beiτz[β]indol-3-yl]-3-oxo-l- propenyl}benzyl-iV,iV-dimethyl-N-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl] ammonium chloride hydrochloride (SN29890); N-4-{(E)-3-[l-(chloromethyl)-5-hydroxy-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo-l-propenyl}benzyl-7V,N-dimethyl-N-[(l-methyl-5-mtro-2- pyrrolyl)methyl]ammonium bromide (SN27829);
N-ό-tS-amino-l-Cchloromethy^-l^-dihydro-SH-benztelindol-S-yy-ό-oxo-l- hexyl-N,JV-dimethyl-JV-{ [1 -methyl-5-(methylsulfonyl)-pyrrol-2-yl]methyl} ammonium salt (SN30155); iV-7- [5 -amino- 1 -(chloromethyl)- 1 ,2-dihydro-3H-benz[e] indol-3 -yl] -7-oxo- 1 - heptanyl-Λζ,N-dimethyl-iV'-{[l-metliyl-5-(methylsulfonyl)-pyrrol-2-yl]metliyl}- ammonium salt (SN30156); N-y-tS-amino-l-Cchloromethy^-l^-dihydro-SH-benzt^indol-S-ylJ-T-oxo-l- heptanyl-N,N-dmiethyl-N-4-(methylsulfonyl)benzylammonium chloride hydrochloride (SN31651);
N-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3/f-benz[e]indol-3-yl]-7-oxo-l- heptanyl-ΛζΛζN-trimethylammoniuin chloride hydrochloride (SN31652); N-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3/f-benz[e]indol-3-yl]-7-oxo-l- heptanyl-N,N-dimethyl-N-4-nitrobenzyl ammonium chloride hydrochloride (SN31653);
N-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3if-benz[e]indol-3-yl]-7-oxo-l- heptanyl-JV-4-cyanobenzyl -Λζ,N-dimethylammonium chloride hydrochloride (SN31666); and iV-4-{(E)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo - 1 -propenyl} benzyl-iV,iV-dimethyl-iV-(4-methylsulfonyl)benzylammonium chloride hydrochloride (SN31630).
According to the present invention, the following intermediates are also preferred: 5-[Bis(te7't-butoxycarbonyl)]amino-l -(chloromethyl)-3-(trifluoroacetyl)-l ,2- dihydro-3H-benz[e]indole (3);
5-[Bis(tert-butoxycarbonyl)]amino-l-(chloromethyl)-3-[(£)-4- (dimethylaminomethyl)cinnamoyl]-l ,2-dihydro-3H-benz[e]indole (4);
E-4-(Dimethylaminomethyl)cinnamic acid (12); 2-(Bromomethyl)-l-methyl-5-nitiOpyrrole (18);
5-(BiOmomethyl)-l-methyl-4-nitroimidazole (23);
2-(Bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole (25); l-(Hydroxymethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3H-benz[e]indole (27);
5-[Bis(tert-butoxycarbonyl)]amino-l-{[(tert-butoxycarbonyl)oxy]methyl}-3- (trifluoroacetyl)- 1 ,2-dihydro-3i7-benz[e]indole (29); 5-[Bis(ter^-butoxycarbonyl)]amino-l-[(fert-butoxycarbonyl)oxy]methyl-3-[(J-t)-
4-(dimethylaminomethyl)cirmamoyl]-l,2-dihydro-3H-benz[e]indole (30);
5 -[Allyl(tert-butoxycarbonyl)amino] -3 -(tert-butoxycarbonyl)- 1 -methyl- 1 ,2- dihydro-3H-benzo[e] indole (34);
5-[Bis(te7't-butoxycarbonyl)]amino-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]- l-methyl-l,2-dihydro-3H-benz|>]mdole (38);
1 -(Chloromethyl)-3-[(E)-4-(dimethylaminoniethyl)cinnamoyl]-5-h.ydroxy- 1 ,2- diliydro-3H-benz[e]indole (42);
5-[Bis(tert-butoxycarbonyl)]amino-l-(chloiOmethyl)-3-[6- (dimethylamino)hexanoyl]-l,2-dihydro-3/i-benz[e]indole (48); and 5-[Bis(fø/^butoxycarbonyl)]ammo-l-(chloromethyl)-3-[7-
(dimethylamino)heptanoyl-l,2-dih.ydiO-3H-benz[e] indole (49).
According to a second aspect of the present invention there is provided a pharmaceutical composition comprising a compound of the first aspect and a pharmaceutically acceptable carrier. According to a third aspect of the present invention there is provided a method of reducing body weight of a subject comprising the administration to a subject of a compound according to the first aspect.
According to a fourth aspect of the present invention there is provided a method of prophylactic or therapeutic treatment of a metabolic or an eating disorder resulting in undesirable body weight gain comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
According to a fifth aspect of the present invention there is provided a method of prophylactic or therapeutic treatment of obesity comprising the administration to a subject requiring such treatment of a compound according to the first aspect. According to a sixth aspect of the present invention there is provided a method of treating hyperglycaemia comprising the administration to a subject requiring such treatment of a compound according to the first aspect. According to a seventh aspect of the present invention there is provided a method of treating hypertriglyceridemia comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
According to an eighth aspect of the present invention there is provided a method of treating Type II diabetes comprising the administration to a subject requiring such treatment of a compound according to the first aspect.
According to a ninth aspect of the present invention there is provided use of a compound according to the first aspect in the manufacture of a medicament for treatment of a condition selected from the group consisting of: excess body weight, metabolic disorders resulting in excess body weight, obesity, high blood glucose and triglycerides and diabetes.
According to a tenth aspect of the present invention there is provided a process for the preparation of a compound according to the first aspect. The process includes but is not limited to the following key steps: 1. The formation of the bisBOC intermediate 3 as shown in Scheme 1, using
DMAP as a catalyst. This reaction proceeds reliably in high yield and avoids possible mixtures of starting material, monoBOC product, and 3, which may be obtained using the literature conditons [Atwell, et al, J. Med. Chem. (1999) 42, 3400-3411]. Similarly formed bisBOC intermediates of general utility include 29 in Scheme 3, 38 in Scheme 4, and 48 and 49 in Scheme 6.
2. The formation of the cinnamide 4 and its quatemisation as shown in Scheme 1 and Scheme 8. Compound 4 is a suitable substrate for quatemisation reactions because of its solubility in relatively nonpolar organic solvents such as THF and EtOAc (from which the quaternary salts, e.g. 5, 6, 7, precipitate or can be made to precipitate on dilution with other suitable solvents such as Et2O)3 and because it contains only one significantly nucleophilic centre (ie. the dimethylamino group) and so generally gives a clean reaction to a single product. Other examples of bisBOC protected intermediates that are also quaternised in a similar manner to 4 are compound 30 in Scheme 3, 38 in Scheme 4, 48 in Scheme 6, and 49 in Scheme 6 and Scheme 7. The quatemisation is generally carried out at room temperature in the dark over several days in the presence of a slight excess of a suitable alkyl halide. ■ In cases where the quaternary salt remains soluble the reaction may be accelerated by the use of inorganic bases (e.g. the use OfK2CO3 in the formation of 50 and 51). 3. The formation of a quaternary salt by direct quaternisation of a compound containing a potentially reactive aniline or phenol functional group. This is illustrated with the synthesis of 10 in Scheme 1 , 37 in Scheme 4, and SN27829 in
Scheme 5.
Unless the context clearly requires otherwise, throughout the description and the claims, the words "comprise", "comprising", and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of "including, but not limited to".
To the extent that any compounds described and claimed herein are described in the relevant prior art and in a relevant manner, they are hereby disclaimed. Brief Description of the Figures
Figure IA. Effect of a single injection of 4.2 μmol/kg SN28127 on body weight, food and water intake on 4 different strains of mice (the data are mean ± SEM).
A single injection of SN28127 induces long term weight loss in four different strains of mice. Four different strains of male mice (C3HeN, a/a, Aw/a, and NZG) aged 50-70 days were housed individually and each animal received a single ip injection of either DMSO (control) or SN28127 (4.2 μmol/kg) in DMSO (drug). Body weight, food and water intake were measured daily for 160 days post injection.
Figure IB. Effect of a single injection of 4.2 μmol/kg SN28127 on plasma glucose, insulin and leptin levels on 4 different strains of mice (the data are mean ± SEM5 C3HeN n = 5 and 4; a/a n = 6 and 6; Ann = 6 and 5; NZG n = 6 and 4) Significant differences determined by Mann Whitney, **, p<0.01, * p<0.05. A single injection of SN28127 did not cause long term diabetes in four different strains of mice. Four different strains of male mice (C3HeN, a/a, Aϊ/γ/a, and NZG) aged 50-70 days were housed individually and each animal received a single ip injection of either DMSO (control) or SN28127 (4.2 μmol/kg) in DMSO (drug). The mice were culled at -160 days post injection and bled from the retro-orbital sinus. Plasma was prepared and used for biochemical measurements and insulin and leptin concentrations were measured using commercial Eliza assay kits.
Figure 2. Effect of a single injection of SN29220 on body weight, food and water intake on C3HeN mice aged ± 50 days (the data are mean for 4 mice). A single injection of SN29220 induces weight loss in C3HeN mice in a dose dependent manner. Male C3HeN mice aged 50-70 days were housed in groups of 4 per cage and each animal received a single ip injection of either DMSO (control) or SN29220 (0.47 μmol/kg, 1.4 μmol/kg, or 4.2 μmol/kg) in DMSO. Body weight food and water intake were monitored and the mice were culled at 60 days post injection.
Figure 3. Effect of 0.42 μmol/kg SN29728 on body weight of C3HeN mice. The mice were aged approximately 50 days at the start of the experiment and received two drug doses, one at day 0 and a repeat injection at day 39 (the data are mean for 4 mice). SN29728 causes reversible weight loss in C3HeN mice. Male C3HeN mice aged 50-70 days were housed in groups of 4 per cage and each animal received a single ip injection of either DMSO (control) or SN29728 (0.42 μmol/kg) in DMSO. The mice received a second injection 39 days after the first injection. The data shows that there was a transient reduction in body weight following the first SN29728 injection and a further decrease in body weight following the second injection. Figure 4. Reversal of high fat diet induced obesity in male C57BL/6J mice with
SN29220. Male C57BL/6J mice were housed in groups of 4 per cage and fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492Ϊ) diet from day 21 (weaning) and then treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 μmol/kg body weight) on day 80 (n = 8 per group).
Figure 5. Reversal of insulin resistance in male C57BL/6J mice fed a high fat diet with SN29220. Male C57BL/6J mice were housed in groups of 4 per cage and fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492Ϊ) diet from day 21 (weaning) and treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 μmol/kg body weight) on day 80. On day 42 post injection, two mice on low fat diet and treated with DMSO, three mice on high fat diet and treated with DMSO, and three mice on high fat diet and treated with 1.4 μmol/kg SN29220 were fasted overnight. The following morning glucose tolerance tests were performed on these mice. Blood glucose was monitored using a glucometer (ACCU-CHEK Advantage, Roche) on a drop of blood collected from the tail tip of each mouse. Immediately after this each mouse received a single ip injection of glucose (2g/kg body weight). At 30 minute intervals over 180 minutes following the glucose injection, blood glucose was monitored using a glucometer on a drop of blood collected from the tail tip of each mouse.
Description of the Preferred Embodiment of the Invention
The present inventors have surprisingly found that a series of indoline compounds are able to produce substantial and lasting weight loss in experimental animals following a single non-toxic dose of the compounds, and that the animals maintain excellent health following this treatment. Some compounds, for example SN29728, have a transient effect and produce further weight loss following a second dose (Figure 3).
The present invention is directed to compounds of general Formula (I):
Figure imgf000010_0001
wherein n is 2 or 3 and when n is 3 the N bears a positive charge which is balanced by a counterion of negative charge
R1 is selected from NH2, OH, NHR, NR2 where R is lower alkyl
R2 is selected from CH2OH5 CH2Cl, CH2Br, CH2I, CH3, CH2OSO2R where R is lower alkyl
R3 is selected from one of the following
Figure imgf000010_0002
wherein m is selected from 1 to 8 p is selected from 1 to 3 X is selected from N or CH the point of attachment is selected from the sites marked 3, 4, 5, or 6 R4 is selected from R (lower alkyl), CH2Ph, CH2(substituted Ph) or CH2(heterocycle or substituted heterocycle); and pharmaceutically acceptable salts thereof. Examples of suitable compounds according to the invention are provided in Table 1, below.
Table 1. Representative Compounds of General Formula (I)
Figure imgf000011_0001
Figure imgf000012_0001
Figure imgf000013_0001
The preparation of compounds of general Formula (I) can be performed in many ways. The starting materials are either known compounds or compounds that may be prepared in analogy with the preparation of similar known compounds. Particularly useful synthetic protocols are outlined in Schemes 1-8, below:
Figure imgf000014_0001
Scheme 2a
Figure imgf000015_0001
11 12
Scheme 2b
Figure imgf000015_0002
I R = Br 13
■— *■ R = NHMe 14
Figure imgf000015_0003
Scheme 2c
O2N OH Br
O2N
N N
Me Me
17 18
Scheme 2d
22
Figure imgf000015_0004
Scheme 2e
MeO2S"^N Me
I R = OH 24
■— *■ R = Br 25 Scheme 3
Figure imgf000016_0001
Scheme4
Figure imgf000017_0001
Scheme 5
Figure imgf000017_0002
Figure imgf000017_0003
42 SN 27829 Scheme 6
Figure imgf000018_0001
n=5SN30155, n=6SN30156 (X=Br or TFA)
Scheme 7
Figure imgf000019_0001
R = N(BOC)2 X = I 53
R = NH2.HCI X = Cl SN31652
Figure imgf000019_0002
R = N(BOC)2 X = Br 54
R = NH2-HCI X = Cl SN31653
Figure imgf000019_0003
I R = N(BOC)2 X = Br 55
"— *■ R = NH2-HCI X = Cl SN31666
Scheme 8
Figure imgf000019_0004
R = NHCH2CH=CH2 56 R= N(BO OCC))22 XX == Br 57 i — R = NH2 SN28125 R = NH2.HCI X = Cl SN31630
'-→- RR R κ - ==- = N(BOC)2 4
Other useful synthesis methods are disclosed in publications for obtaining structurally similar compounds. To reiterate, any discussion of the prior ait throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field. The following representative publications are incorporated herein by reference:
Atwell, Graham J.; Milbank, Jared J. B.; Wilson, William R.; Hogg, Alison; Denny, William A. 5-Amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indoles: Relationships between Structure and Cytotoxicity for Analogues Bearing Different DNA Minor Groove Binding Submits. J. Med. Chem. (1999), 42(17), 3400-3411. This document describes the synthesis of 1, 2, and monoBOC analogue of 3. The latter is used to prepare amides like 4 from which the BOC is removed, analogous to the methodology described herein. Yang, Shangjin; Denny, William A. A New Short Synthesis of 3 -Substituted 5-
Amino- l-(chloromethyl)-l,2-dihydro-3H-benzo[e]indoles (Amino-CBIs). J. Org. Chem. (2002), 67(25), 8958-8961. This document describes the synthesis of 33, 43, and the deprotection of 43 and formation of amides from this, analogous to the methodology described herein. Boger, Dale L.; McKie, Jeffrey A. An Efficient Synthesis of 1,2,9,9a-
Tetrahydrocyclopropa[c]benz[e]indol-4-one CBI: An Enhanced and Simplified Analog of the CC- 1065 and Duocarmycin Alkylation Submits. J. Org. Chem. (1995) 60(5), 1271-1275. This document describes the synthesis of 41, and, in references cited therein, the deprotection of 41 and formation of amides from this. These publications will enable the skilled person to produce compounds within the scope of Formula (I) by utilising common general knowledge and/or the synthesis methods disclosed.
For use in medicine, the salts of the compounds of this invention refer to nontoxic "pharmaceutically acceptable salts." Other salts may, however, be useful in the preparation of compounds according to this invention or of their pharmaceutically acceptable salts. Suitable pharmaceutically acceptable salts of the compounds include the counterion of the quaternary ammonium species, which may be formed during the quaternisation reaction, or by ion exchange chromatography using a resin bearing the conjugate base of a pharmaceutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Other suitable pharmaceutically acceptable salts of the compounds include acid addition salts which may, for example, be formed by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid.
Furthermore, where the compounds of the invention carry an acidic moiety, they may be prepared as zwitterionic species, or as suitable pharmaceutically acceptable salts of the acidic moiety, these salts may include alkali metal salts, e.g. sodium or potassium salts; alkaline earth metal salts, e.g. calcium or magnesium salts; and salts formed with suitable organic ligands, e.g. quaternary ammonium salts. Thus, representative pharmaceutically acceptable salts include the following: acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, calcium edetate, camsylate, carbonate, chloride, clavulanate, citrate, dihydro chloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, oleate, pamoate (embonate), palmitate, pantothenate, phosphate/diphosphate, polygalacturonate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide and valerate.
The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs will be functional derivatives of the compounds which are readily convertible in vivo into the required compound. Thus, in the methods of treatment of the present invention, the term "administering" shall encompass the treatment of the various disorders described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to the patient. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in "Design of Prodrugs", Ed. H. Bundgaard, Elsevier, 1985, incorporated in its entirety herein by reference.
Where the compounds according to this invention have at least one chiral centre, they may accordingly exist as enantiomers. Where the compounds possess two or more chiral centres, they may additionally exist as diastereomers. It is to be understood that all such isomers and mixtures thereof are encompassed within the scope of the present invention. Furthermore, some of the crystalline forms for the compounds may exist as polymorphs and as such are intended to be included in the present invention. In addition, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are also intended to be encompassed within the scope of this invention. The invention also encompasses active metabolites of a compound of the invention. Pharmaceutical Compositions
The present invention also includes within its scope pharmaceutical compositions comprising, as an active ingredient, at least one compound of the invention or a pharmaceutically acceptable salt or prodrug or hydrate thereof together with a pharmaceutically acceptable carrier or diluent.
Pharmaceutical compositions containing a compound of the invention of the present invention may be prepared by conventional techniques, e.g. as described in "Remington: The Science and Practise of Pharmacy", 19th Edn., 1995. The compositions may appear in conventional forms, for example capsules, tablets, aerosols, solutions, suspensions or topical applications.
Typical compositions include a compound of the invention or a pharmaceutically acceptable basic addition salt or prodrug or hydrate thereof, associated with a pharmaceutically acceptable excipient which may be a carrier or a diluent or be diluted by a carrier, or enclosed within a carrier which can be in the form of a capsule, sachet, paper or other container. In making the compositions, conventional techniques for the preparation of pharmaceutical compositions may be used. For example, the active compound will usually be mixed with a carrier, or diluted by a carrier, or enclosed within a carrier which may be in the form of an ampoule, capsule, sachet, paper, or other container.
When the carrier serves as a diluent, it may be solid, semi- solid, or liquid material, which acts as a vehicle, excipient, or medium for the active compound. The active compound can be adsorbed on a granular solid container for example in a sachet. Some examples of suitable carriers are water, salt solutions, alcohols, polyethylene glycols, polyhydroxyethoxylated castor oil, peanut oil, olive oil, gelatine, lactose, terra alba, sucrose, dextrin, magnesium carbonate, sugar, cyclo dextrin, amylose, magnesium stearate, talc, gelatine, agar, pectin, acacia, stearic acid or lower alkyl ethers of cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglyceiϊdes and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone. Similarly, the carrier or diluent may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with a wax. The formulations may also include wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavouring agents. The formulations of the invention may be formulated so as to provide quick, sustained, or delayed release of the active ingredient after administration to the patient by employing procedures well known in the art.
The pharmaceutical compositions can be sterilised and mixed, if desired, with auxiliary agents, emulsifiers, salt for influencing osmotic pressure, buffers and/or colouring substances and the like, which do not deleteriously react with the active compounds.
The route of administration may be any route, which effectively transports the active compound of the invention to the appropriate or desired site of action, such as oral, nasal, pulmonary, buccal, subdermal, intradermal, transdermal or parenteral e.g. rectal, depot, subcutaneous, intravenous, intraurethral, intramuscular, intranasal, ophthalmic solution or an ointment, the oral route being preferred.
If a solid carrier is used for oral administration, the preparation may be tabletted, placed in a hard gelatin capsule in powder or pellet form or it can be in the form of a troche or lozenge. If a liquid carrier is used, the preparation may be in the form of a syrup, emulsion, soft gelatin capsule or sterile injectable liquid such as an aqueous or non-aqueous liquid suspension or solution.
For nasal administration, the preparation may contain a compound of the invention dissolved or suspended in a liquid carrier, in particular an aqueous carrier, for aerosol application. The carrier may contain additives such as solubilizing agents, e.g. propylene glycol, surfactants, absorption enhancers such as lecithin (phosphatidylcholine) or cyclo dextrin, or preservatives such as parabenes.
For parenteral application, particularly suitable are injectable solutions or suspensions, preferably aqueous solutions with the active compound dissolved in polyhydroxylated castor oil. Tablets, dragees, or capsules having talc and/or a carbohydrate carrier or binder or the like are particularly suitable for oral application. Preferable carriers for tablets, dragees, or capsules include lactose, corn starch, and/or potato starch. A syrup or elixir can be used in cases where a sweetened vehicle can be employed. The compounds of Formula (I) may be administered to a mammal, especially a human in need of such treatment, prevention, elimination, alleviation or amelioration of the various disorders such as metabolic disorders, excessive or undesirable weight gain, obesity Type II diabetes, hyperglycaemia, hypertriglyceridemia, disorders of food intake and related conditions. In particular, the compounds of Formula (I) are contemplated to be useful for the prevention or treatment of obesity. Furthermore, the compounds of Formula (I) may be useful for blood glucose lowering, prevention of hyperglycemia, delaying the progression of impaired glucose tolerance to Type II diabetes, delaying the progression of non- insulin requiring Type II diabetes to insulin requiring Type II diabetes, or appetite regulation. The mammal to be treated with a compound of Formula (I) is preferably a human, but may also be a domesticated animal, e.g. household pet or farmyard animal, and non-domesticated animal such as wildlife.
The compounds of the invention are effective over a wide dosage range. From the experimental studies described herein the representative compounds typically showed activity from about 300 μg/kg to about 3000 μg/kg. For example, in the treatment of adult humans, dosages from about 20mg to about 250 mg per day may be used and may be divided into multiple doses. The dosage need not be daily and the requirement for administration would be gauged by the response of an individual. In choosing a regimen for patients it may frequently be necessary to begin with a higher dosage and when the condition is under control to reduce the dosage. The exact dosage will depend upon the mode of administration, on the therapy desired, form in which administered, the subject to be treated and the body weight of the subject to be treated, and the preference and experience of the physician or veterinarian in charge.
Generally, the compounds of the present invention are dispensed in unit dosage form comprising from about 20 mg to about 50 mg of active ingredient together with a pharmaceutically acceptable carrier per unit.
The invention furthermore relates to the use of a compound according to the present invention for the preparation of a medicament for use in the treatment of excessive or undesirable weight gain, obesity and associated conditions in a regimen which additionally comprises treatment with another anti-obesity or anti-weight gain agent. For example the compounds of the present invention may be administered in conjunction with a known anti-diabetic agent. In the present context the expression "antidiabetic agent" includes compounds for the treatment and/or prophylaxis of insulin resistance and diseases wherein insulin resistance is the pathophysiological mechanism. The compounds may also be administered in conjunction with a hypoglycaemic agent, another anti-obesity agent or an antilipidemic agent.
Preferred embodiments of the invention will now be described with reference to the accompanying, non-limiting, examples.
Examples
The following examples describe the invention in greater detail and are intended to illustrate the invention, but not to limit it. AU compounds were identified by a variety of methods including nuclear magnetic resonance spectroscopy, mass spectrometry and in some cases, elemental analysis. Nuclear Magnetic Resonance (400 MHz NMR) data are reported in parts per million downfϊeld from tetramethylsilane (TMS). Mass spectra data are reported in mass/charge (m/z) units. Unless otherwise noted, the materials used in the example were obtained from readily available commercial sources or synthesised by standard methods known to those skilled in the art. With reference to Scheme 1:
Example 1: Synthesis of 5-[bis(te/^butoxycarbonyl)]amino-l-(cbloromethyl)-3- (trifluoroacetyl)-l,2-dihydro-3JΪ-benz[e]indole (3)
A solution of l-(chloromethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro-3//'- benz[ejindole [Atwell et al., J. Med Chem. (1999) 42, 3400-3411] (1) (978 mg, 2.72 mmol) in THF (30 mL) and MeOH (5 mL) was hydrogenated over PtO2-XH2O (Acros, Pt content 79-84%, 0.24 g) at 40 psi for 40 min. The mixture was filtered through Celite, the filtrate was evaporated, and the residue was purified by column chromatography (EtOAcrpetroleum ether 1:2) to give 5-amino-l-(chloromethyl)-3-(trifluoroacetyl)-l,2- dihydro-3H-benz[e]indole (2) as a cream solid (0.64 g, 71%), which was used directly in the next step. The aniline 2 (0.64 g, 1.95 mmol) was dissolved in dioxane (50 mL) with di-tert-butyl dicarbonate (1.5 g, 6.9 mmol) and DMAP (5 mg) and the solution stirred at reflux under nitrogen for 16 h. More di-tert-buty\ dicarbonate (1.2 g, 5.5 mmol) was added and the solution stirred at reflux for a further 2 h.
The solvent was evaporated and the residue purified by column chromatography (EtOAc:petroleum ether 4:1) to give 5-[Bis(terr-butoxycarbonyl)]amino-l-
(chloromethyl)-3-(trifluoroacetyl)-l,2-diliydro-3i7-benz[e]indole 3 as a cream solid (0.94 g, 91%). A sample was recrystallised from PhH-petroleum ether as white prisms: m.p. 181-182 °C; 1H NMR (CDCl3) δ 8.37 (s, 1 H), 7.87 (d, J= 8.3 Hz, 1 H), 7.80 (d, J = 8.1 Hz, 1 H)5 7.59 (dt, J= 7.6, 1.2 Hz, 1 H), 7.53 (dt, J= 7.6, 1.3 Hz, 1 H)5 4.64 (d, J= 11.4 Hz5 1 H)5 4.45 (dt, J= 11.4, 8.5 Hz5 1 H)5 4.26-4.18 (m, 1 H)5 3.97 (dd, J= 11.55 3.2 Hz5 1 H)5 3.57 (dd, J= 11.5, 9.4 Hz5 1 H)5 1.38 (s, 9 H)5 1.35 (s, 9 H). Anal. CaIc. for C25H28ClF3N2O5: C5 56.77; H5 5.34; N5 5.30. Found: C5 56.63; H, 5.49; N5 5.23. Example 2: Synthesis of 5-[bis(tert-butoxycarbonyl)]amino-l-(chloromethyl)-3- [(£)-4-(dimethylaminomethyl)cinnamoyl]-l,2-dihydro-3JHr-benz[g]indole (4)
A solution OfCs2CO3 (1.4 g, 4.3 mmol) in H2O (30 mL) was added to a solution of 3 (440 mg, 0.83 mmol) in THF (40 mL), and the two-phase mixture was stirred vigorously at 20 °C for 8 h.
The THF was evaporated and the residue was diluted with aqueous NaCl and extracted with EtOAc (x3). The extracts were washed with aqueous NaCl5 dried (Na2SO4), and evaporated to provide crude 5-[bis(fert-butoxycarbonyl)]amino-l- (chloromethyl)-l,2-dihydro-3H-benz[e]indole as a yellow oil, which was used directly in the next step. Oxalyl chloride (0.44 mL, 5.04 mmol) and then DMF (1 drop) were added to a suspension of E-4-(dimethylaminomethyl)cinnamic acid (256 mg, 1.24 mmol) in dry CH3CN (20 mL) and the mixture stirred at 20 °C. After 5.5 h the mixture was evaporated. The amine prepared above was dissolved in THF (40 mL) and added to the acid chloride, followed by the addition OfEt3N (0.52 mL, 3.73 mmol). The resulting light brown suspension was stirred at 20 0C for 45 min. Trial reactions on a small scale using the same reagents showed that the reaction mixture contained some methyl E-4-(dimethylaminomethyl)-cinnamate, which co-eluted with the desired product. To convert this byproduct to the acid and aid purification, a solution of KOH (0.58 g, 10 mmol) in H2O (6 mL) was added, and the mixture stirred at 20 0C for a further 12 h.
The THF was evaporated, the aqueous residue was extracted with EtOAc (x3), and the extracts were washed with aqueous NaCl, dried (Na2SO4), and evaporated. Purification of the residue by column chromatography (EtOAc then EtOAc:MeOH 4:1) gave 5-[Bis(ter/1-butoxycarbonyl)]amino-l-(cliloromethyl)-3-[(£')-4- (dimethylaminomethyl)cinnamoyl]-l,2-diliydro-3H-benz[e]indole 4 as a yellow foam (480 ing, 77%): 1H NMR (CDCl3) δ 8.58 (br s, 1 H)5 7.87 (d, J= 15.3 Hz5 1 H)5 7.82 (d, J= 8.4 Hz5 1 H)5 7.76 (d, J= 8.2 Hz, 1 H), 7.58 (d, J= 8.0 Hz, 2 H) 7.53 (t, J= 7.5 Hz5 1 H), 7.45 (t, J= 7.6 Hz, 1 H)5 7.37 (d, J= 8.0 Hz5 2 H)5 6.90 (d, J= 15.3 Hz5 1 H)5 4.59- 4.53 (m, 1 H)5 4.46 (t, J= 9.5 Hz, 1 H), 4.24-4.14 (m, 1 H), 3.99 (dd, J= 11.3, 2.9 Hz, 1 H), 3.53 (t, J= 10.8 Hz, 1 H), 3.50 (s, 2 H), 2.29 (s, 6 H), 1.36 (s, 18 H); 13C NMR δ
164.4. 151.8, 151.7, 144.0, 141.1, 140.8, 137.3, 133.9, 129.7, 129.6, 128.4, 128.2, 127.3,
125.3. 123.9, 123.6, 122.7, 118.3, 118.0, 82.9, 63.8, 53.1, 46.1, 45.2, 42.7, 27.8; HRMS (FAB, 35Cl) m/z 620.29015 (M+H), C35H43ClN3O5 requires 620.28912.
Example 3: Synthesis of 5-amino-l-(chloromethyl)-3-[(£)-4- (dimethylaminomethyl)-cmnamoyl]-l,2-dihydro-3JΪ-benz[e]mdoIe (SN28125)
. A solution of 4 (188 mg, 0.30 mmol) in dioxane (15 mL) was saturated with HCl gas and stirred at 20 0C for 1 h, during which time the HCl salt of the product precipitated. Aqueous NaHCO3 was added and the mixture was extracted with EtOAc (x3). The extracts were dried (Na2SO4) and evaporated, and the residue was purified by column chromatography (EtOAc then EtOAc:MeOH 9:1 then 4:1).
The product was recrystallised from PhH giving 5-Amino-l-(chloromethyl)-3- [(E)-4-(dimethylaminomethyl)cinnamoyl]-l,2-dihydro-3H-benz[e]indole SN28125 as a yellow solid (91 mg, 71%): m.p. 230-240 0C (dec); 1H NMR (CDCl3) δ 8.04 (br s, 1 H), 7.86 (d, J= 15.3 Hz, 1 H), 7.81 (d, J= 8.5 Hz, 1 H), 7.68 (d, J= 8.3 Hz, 1 H), 7.57 (d, J = 8.1 Hz, 2 H) 7.50 (t, J= 7,3 Hz, 1 H), 7.39-7.34 (m, 3 H), 6.91 (d, J= 15.2 Hz, 1 H), 4.51 (d, J= 10.6 Hz, 1 H)5 4.37 (t, J= 9.5 Hz, 1 H), 4.32 (s, 2 H), 4.09-4.02 (m, 1 H), 3.95 (dd, J= 11.2, 2.8 Hz, 1 H), 3.47 (s, 2 H), 3.43 (t, J= 11.1 Hz, 1 H)5 2.27 (s, 6 H); 13C NMR δ 164.6, 144.2, 143.7, 142.1, 141.2, 133.9, 130.1, 129.6, 128.1, 127.2, 123.3, 122.9, 122.1, 121.1, 118.3, 114.6, 101.4, 64.0, 53.3, 46.3, 45.4, 42.6. Anal. CaIc. for C25H26ClN3O: C, 71.50; H, 6.24; N, 10.01. Found: C, 71.66; H, 6.18; N, 9.93. Example 4: Synthesis of N-4-((£)-3-{5-[bis(tert-butoxycarbonyl)]amino-l- (chloromethy^-l^-dihydro-SJϊ-benzJeJindol-S-ylJ-S-oxo-l-propeny^benzyl-iVyV- dimethyl-Λr-[(l-methyl-5-nitro-2-pyrrolyl)methyl]ammonium bromide (5)
A solution of 4 (181 mg, 0.29 mmol) and 17 (76.7 mg, 0.35 mmol) in THF (6 mL) was stirred in the dark at 20 °C for 3 days, giving a fine yellow precipitate. The precipitate was allowed to settle and the supernatant was decanted off.
The remaining solid was triturated several times with CH3CN, then filtered off and dried to give N-4-((E)-3-{5-[bis(ter^butoxycarbonyl)]amiiio-l-(chloiOmethyl)-l;2- dπiydiO-3/J-benz[e]indol-3-yl}-3-oxo-l-piOpenyl)benzyl-N,N-dimethyl-N-[(l-methyl-5- nitiO-2-pyrrolyl)memyl]ammonium bromide 5 as a pale yellow solid (166 mg, 68%): m.p. 221 0C (dec); 1H NMR tø-DMSO) δ 8.40 (s, 1 H), 8.03 (d, J= 8.3 Hz, 1 H), 7.98 (d, J= 8.0 Hz5 2 H), 7.76 (d, J= 15.4 Hz, 1 H), 7.68 (d, J= 8.3 Hz, 1 H), 7.64 (d, J= 8.3 Hz, 2 H), 7.60 (t, J= 7.6 Hz, 1 H), 7.53 (t, J= 7.5 Hz, 1 H), 7.38 (d, J= 4.4 Hz, 1 H), 7.35 (d, J= 16 Hz, 1 H), 6.70 (d, J= 4.5 Hz, 1 H), 4.83 (s, 2 H), 4.66 (s, 2 H)5 4.65-4.47 (m, 3 H), 4.09-3.99 (m, 2 H), 4.02 (s, 3 H), 2.95 (s, 6 H), 1.37 (s, 9 H), 1.36 (s, 9 H). Anal. CaIc. for C41H49BrON5O7: C, 58.68; H, 5.88; N, 8.35; Br, 9.52. Found: C, 58.91; H5 5.77; N, 8.23; Br, 9.74.
Example 5: Synthesis of iV-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3Hr- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-N^V-dimethyl-Ar-[(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN28127) Dioxane (3 mL) saturated with HBr gas was added to a suspension of 5 (144 mg,
0.17 mmol) in dioxane (6 mL); the mixture was stirred in the dark at 20 ° C for 12 h.
Excess HBr was removed under reduced pressure, then the solid was filtered off and dried to give N-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol- 3 -yl] -3 -oxo - 1 -propenyl} benzyl- JV5 JV-dimethy 1- JV- [(I -methyl-5 -nitro -2- pyrrolyl)methyl]ammonium bromide hydrobromide SN28127 as a yellow solid (120 mg, 97%): m.p. 170-180 °C (dec); 1H NMR (J6-DMSO) δ 8.20 (br s, 1 H), 8.04 (d, J= 8.5 Hz, 1 H), 7.97 (d, J= 8.2 Hz5 2 H)5 7.86 (d, J= 8.4 Hz, 1 H), 7.73 (d, J= 15.3 Hz, 1 H), 7.64 (d, J= 8.2 Hz, 2 H)5 7.54 (t, J= 7.5 Hz, 1 H), 7.41 (t, J= 7.6 Hz, 1 H)5 7.37 (d, J= 4.5 Hz, 1 H)5 7.35 (d, J= 16 Hz5 1 H)5 6.70 (d, J= 4.5 Hz5 1 H), 4.84 (s, 2 H)5 4.67 (s, 2 H), 4.55-4.46 (m, 2 H), 4.31-4.26 (m, 1 H), 4.03 (s, 3 H)5 3.99 (dd, J = 11.1, 2.7 Hz, 1 H), 3.85 (dd, J= 10.4, 8.1 Hz, 1 H)5 2.96 (s, 6 H); HRMS (FAB, 35Cl) m/z 558.22205, C31H33ClN5O3 requires 558.22719.
HPLC analysis indicated the presence of 0.3% SN28125. Example 6: Synthesis of JV-4-((£)-3-{5-[bis(tø^-butoxycarbonyl)]amino-l- (chloromethy^-l^-dihydro-Siϊ-benzl^indol-S-ylJ-S-oxo-l-propeny^benzyl-JY^/V- dimethyl-JV- [(l-methyl-4-nitro-5-imidazolyl)methyl] ammonium bromide (6)
A solution of 4 (52.9 mg, 0.085 mmol) and 22 (22.5 mg, 0.10 mmol) in EtOAc (1.5 mL) was allowed to stand in the dark at 20 °C for 3 days. Et2O (3 mL) was added dropwise giving a slightly cloudy mixture that was allowed to stand for 12 h. The precipitated solid was filtered off and dried to give JV-4-((£)-3-{5-[bis(fe7t- butoxycarbonyl)]amino-l -(chloro methyl)- 152-diliydro-3H-benz[e]indol-3-yl} -3-oxo-l - propenyl)benzyl-JV,JV-dύnethyl-N-[(l-methyl-4-nitro-5-imidazolyl)methyl]amrnonium bromide 6 as a yellow solid (55 mg, 77%). 1H NMR (W6-DMSO) δ 8.40 (s, 1 H), 8.15 (s, 1 H), 8.03 (d, J= 8.4 Hz, 1 H)5 7.99 (d, J= 8.0 Hz, 2 H), 7.76 (d, J= 15.4 Hz, 1 H), 7.68 (d, J= 8.2 Hz, 2 H), 7.61 (t, J= 7.4 Hz, 1 H), 7.54 (t, J= 7.6 Hz, 1 H), 7.37 (d, J= 15.5 Hz, 1 H), 5.07 (tar s, 2 H), 4.77 (s, 2 H), 4.68-4.55 (m, 2 H), 4.53-4.47 (m, 1 H), 4.09-3.99 (m, 2 H), 3.88 (s, 3 H), 3.01 (s, 6 H), 1.37 (s, 9 H), 1.36 (s, 9 H); HRMS (FAB, 35Cl) m/z 759.32810 (M+), C40H48ClN6O7 requires 759.32730.
Example 7: Synthesis of A7-4-{(jE)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3iϊ- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-ΛyV-dimethyl-iV-[(l-methyl-4-nitro-5- imidazolyl)methyl] ammonium bromide hydrobromide (SN28126) Dioxane (1 mL) saturated with HBr gas was added to a suspension of 6 (54 mg,
0.064 mmol) in dioxane (2.5 mL) and the mixture stirred in the dark at 20 0C for 3.5 h. Excess HBr was removed under reduced pressure, then the solid was filtered off and dried to give N-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]- 3 -oxo- 1 -propenyl} benzyl- JV,JV-dimethy 1-JV- [(I -methyl-4-nitro -5 - imidazolyl)methyl]ammonium bromide hydrobromide SN28126 as a yellow powder (47 mg, 100%): m.p. 170-180 °C (dec); 1HNMR (J6-DMSO) δ 8.25 (tar s, 1 H), 8.15 (s, 1 H), 8.04 (d, J= 8.4 Hz, 1 H), 7.99 (d, J= 8.1 Hz, 2 H), 7.89 (d, J= 8.4 Hz, 1 H), 7.74 (d, J= 15.4 Hz, 1 H), 7.69 (d, J= 8.2 Hz, 2 H), 7.56 (t, J= 7.6 Hz, 1 H), 7.44 (t, J= 7.5 Hz, 1 H), 7.36 (d, J= 15.4 Hz5 1 H), 5.08 (tar s, 2 H), 4.80 (s, 2 H), 4.57-4.47 (m, 1 H), 4.34- 4.27 (m, 1 H), 3.99 (dd, J= 11.0, 2.8 Hz, 1 H), 3.91-3.85 (m, 4 H)5 3.02 (s, 6 H). Anal. CaIc. for C30H32BrClN6O3 -HBr-4H2O: C, 45.44; H, 5.21; N5 10.60. Found: C5 45.01; H, 4.88; N, 10.23. HPLC analysis indicated the presence of 1% SN28125. Example 8: Synthesis of iV-4-((jE)-3-{5-[bis(to^-butoxycarbonyl)]amino-l- (chloromethyI)-l,2-dihydro-3Jjr-benz[e]indol-3-yl}-3-oxo-l-propenyl)benzyl-iV^V- dimethyl-JV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide (7)
A solution of 4 (159 mg, 0.256 mmol) and 25 (97 mg, 0.385 mmol) in THF (4 mL) was stirred in the dark at 20 °C for 3 days.
The precipitate was filtered off and triturated with Et2O to give N-4-((E)-3-{5- [bis(fø;t-butoxycarbonyl)]amino-l-(chloromethyl)-l,2-dihydro-3H-benz[β]indol-3-yl}- 3-oxo-l-piOpenyl)benzyl-iV,7V-dimethyl-iV:-[(l-methyl-5- methylsulfonyl-2- pyrro Iy l)methyl] ammonium bromide 7 (206 mg, 92%) as a pale yellow solid: m.p. 182- 185 °C (dec); 1H NMR (J6-DMSO) δ 8.40 (s, 1 H), 8.03 (d, J= 8.3 Hz, 1 H), 7.98 (d, J = 8.0 Hz, 2 H), 7.77 (d, J= 15.4 Hz5 1 H), 7.72-7.64 (m, 3 H)5 7.61 (td, J= 7.5, 1.1 Hz, 1 H), 7.54 (td, J= 7.6, 1.0 Hz, 1 H), 7.35 (d, J= 11.5 Hz, 1 H), 6.92 (d, J= 4.1 Hz, 1 H), 6.64 (d, J= 4.1 Hz, 1 H), 4.77 (s, 2 H)5 4.65 (s, 2 H), 4.62-4.54 (m, 2 H), 4.53-4.46 (m, 1 H), 4.07 (dd, J= 11.3, 3.3 Hz5 1 H)5 4.01 (dd, J= 11.2, 6.4 Hz, 1 H)5 3.97 (s, 3 H), 3.32 (s, 3 H)5 2.92 (s, 6 H)5 1.37 (s, 9 H)5 1.35 (s, 9 H). Anal. CaIc. for C42H52BrClN4O7S: C, 57.83; H, 6.01; N5 6.42. Found: C5 57.55; H, 6.01; N, 6.34.
Example 9: Synthesis of 7V-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-ΛVV-dimethyl-iV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) Dioxane (2 mL) saturated with HBr gas was added to a solution of 7 (40 mg, 0.046 mmol) in dioxane (1 mL) and the mixture was stirred in the dark at 20 °C for 15 h. Excess HBr was removed under reduced pressure, then the solid was filtered off and triturated with Et2O to give iV-4-{(E)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H- benz[e] indol-3 -yl] -3 -oxo- 1 -propenyl} benzyl-iV, JV-dimethyl-iV-[(l -methyl-5 - methylsulfonyl-2-pyrrolyl)methyl]ammonium bromide hydrobromide SN29220 as a yellow solid (28 mg, 80%): m.p. 178-182 0C (dec); 1H NMR tø-DMSO) δ 8.07 (d, J= 8.5 Hz, 1 H), 8.01 (s, 1 H)5 7.96 (d, J= 8.2 Hz5 2 H), 7.80 (d, J= 8.4 Hz5 1 H), 7.71 (d, J = 15.4 Hz5 1 H)5 7.65 (d, J= 8.2 Hz5 2 H), 7.50 (t, J= 7.5 Hz5 1 H), 7.34 (t, J= 7.5 Hz5 1 H), 7.33 (d, J= 15.4 Hz5 1 H)5 6.92 (d, J= 4.1 Hz5 1 H), 6.65 (d, J= 4.1 Hz5 1 H), 4.78 (s, 2 H), 4.64 (s, 2 H)5 4.53-4.42 (m5 2 H), 4.25-4.16 (m, 1 H)5 4.00-3.93 (m, 1 H)5 3.95 (S5 3 H)5 3.79 (dd, J= 11.1, 8.1 Hz, 1 H), 3.28 (s, 3 H), 2.91 (s, 6 H); HRMS (FAB, 35Cl) m/z 591.2213 (M+l), C32H36ClN4O3S requires 591.2197.
N-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo-l-propeiiyl}benzyl-A/5iV-dimethyl-N-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl]ammonium bromide hydrobromide SN29220 was also obtained as a mixture of bromide and trifluoro acetate salts by deprotection of iV-4-((£)-3-{5-[bis(ført- butoxycarbonyl)]amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl}-3-oxo-l- propenyl)benzyl-iV,iV-dimethyl-JV-[(l -methyl-5- methylsulfonyl-2- pyrro Iy l)methyl] ammonium bromide 7 with TFA/CH2C12 (1:1). Example 10: Synthesis of N-4-((JE)-3-{5-[bis(tert-butoxycarbonyl)]amino-l- (chloromethy^-l^-dihydro-SjfiT-benz^Jindol-S-ylJ-S-oxo-l-propeny^benzyl-iVjN^/V- trimethyl-ammonhim iodide (8)
A solution of 4 (180 mg, 0.29 mmol) and methyl iodide (22 μL, 0.35 mmol) in EtOAc (2 mL) was allowed to stand in the dark at 20 °C for 16 h. Et2O was added dropwise, causing an oil to separate, and the supernatant was decanted off. The remaining oil was triturated with EtOAc, and the resulting solid was filtered off to give iV-4-((£)-3-{5-[bis(tert-butoxycarbonyl)]amino-l-(chloromethyl)-l,2-dihydro-3H- benz[e]indol-3-yl}-3-oxo-l-propenyl)beiizyl-N",N,N-trimethylanimonium iodide 8 (201 mg, 91%): m.p. >320 0C; 1H NMR (J6-DMSO) δ 8.40 (s, 1 H), .8.03 (d, J= 8.3 Hz, 1 H),
7.97 (d, J= 8.0 Hz, 2 H), 7.75 (d, J= 15.4 Hz, 1 H), 7.68 (d, J= 8.5 Hz, 1 H), 7.63-7.58 (m 3 H), 7.53 (t, J= 7.6 Hz, 1 H), 7.34 (d, J= 15.4 Hz, 1 H), 4.67-4.47 (m, 5 H), 4.08-
3.98 (m, 2 H), 3.05 (s, 9 H), 1.37 (s, 9 H), 1.36 (s, 9 H). Anal. CaIc. for C36H45ClIN3O5-H2O: C, 55.43; H, 6.07; N, 5.39. Found: C, 55.66; H, 5.99; N, 5.33. Example 11: Synthesis of N-(4-{(£)-3-[5-amino-l-(chIoromethyI)-l,2-dihydro-3JΪ- benz[e]indoI-3-yl]-3-oxo-l-propenyl}benzyl)-Λy\yV-trimethylammonium chloride hydrochloride (SN29728)
Dioxane (2 mL) saturated with HCl gas was added to a solution of 8 (175 mg, 0.23 mmol) in dioxane (4 mL). A precipitate began to form within a few minutes. The mixture was stirred at 20 0C for 16 h, and the excess HCl removed under reduced pressure. The solid was filtered off, washed with dioxane, and dried to give SN29728 as a yellow-orange solid (122 mg): m.p. >310 °C; 1H NMR (J6-DMSO) δ 8.15 (br s, 1 H), 8.07 (d, J= 8.5 Hz, 1 H), 7.96 (d, J= 8.2 Hz, 2 H), 7.85 (d, J= 8.3 Hz, 1 H), 7.73 (d, J= 15.4 Hz, 1 H), 7.61 (d5 J= 8.2 Hz, 2 H), 7.53 (d, J= 7.6 Hz, 1 H), 7.39 (t, J= 7.6 Hz, 1 H), 7.33 (d, J= 15.4 Hz, 1 H), 4.57 (s, 2 H), 4.54-4.45 (m, 2 H), 4.30-4.23 (m, 1 H), 3.99 (dd, J= 11.0, 3.0 Hz, 1 H), 3.84 (dd, J= 11.0, 7.8 Hz, 1 H), 3.07 (s, 9 H). Microanalysis indicated a mixture of chloride and iodide counterions. The mixed salt (110 mg) was dissolved in MeOH (2 mL) and purified by ion exchange chromatography (Biorad AG- 1X4 resin in Cl" form), eluting with MeOH. The yellow product-containing fractions were concentrated to a small volume and diluted with EtOAc to give N-(4-{(E)-3-[5- amino- 1 -(chloromethyl)- 1 ,2-dihydro-3/i-benz[e] indol-3 -yl] -3 -oxo- 1 -propenyl} benzyl)- N,Λζ N-trimethylammonium chloride hydrochloride SN29728 as a yellow solid (83 mg, 79%). Anal. CaIc. for C26H30Cl3N3O-H2O-1Z2EtOAc: C, 59.11; H, 6.38; N, 7.39; Cl, 18.69. Found: C, 58.90; H, 6.41; N, 7.77; Cl, 18.24. Example 12: Synthesis of 5-[bis(te/Y-butoxycarbonyl)]amino-l-(chloromethyl)-3- {(£)-3-[4-({methyl[(l-methyl-5-nitro-2-pyrrolyl)methyl]amino}methyl)phenyl]-2- propenoyl}-l,2-dihydro-3iϊ-benz[e]indole (9) A solution of 3 (60 nig, 0.114 mmol) and Cs2CO3 (153 nig, 0.456 mmol) in CH2Cl2 (5 mL) and MeOH (5 niL) was stirred for 15 min. CH2Cl2 (20 niL) and H2O were added and the organic layer was separated.
The organic layer was washed with H2O, brine, and dried (Na2SO4), and then evaporated to provide crude 5-[bis(tert-butoxycarbonyl)]amino-l-(chloromethyl)-l,2- dihydro-3H-benz[e]indole as a yellow oil (49 mg), which was used directly in the next step.
Oxalyl chloride (187 mg, 1.54 mmol) and then DMF (1 drop) were added to a suspension of 16 (76 mg, 0.171 mmol) in dry CH3CN (3 mL). The mixture was stirred for 2 h and then evaporated under reduced pressure. The amine prepared above was dissolved in THF (3 mL) with Et3N (58 mg, 0.57 mmol) and the resulting mixture was added to the acid chloride in THF (2 mL). The mixture was stirred for 2 h., the solvents were removed under reduced pressure, and EtOAc and H2O were added to the residue.
The EtOAc extract was washed with brine and dried (Na2SO4). Column chromatography (petroleum ether.EtOAc 7:3 then 3:2) followed by trituration with petroleum ether gave 5-[Bis(tert-butoxycarbonyl)]amino-l-(chloromethyl)-3-{(E)-3-[4- ( {methyl[(l -methyl-5 -nitro-2-pyrrolyl)methyl] amino } methyl)phenyl] -2-propenoyl} -1,2- diliydro-3Jf-benz[e]indole 9 (53 mg, 63%) as ayellow powder: m.p. 110-115 0C (dec); 1H NMR (CDCl3) δ 8.57 (s, 1 H), 7.88 (d, J= 15.3 Hz, 1 H), 7.82 (d, J= 8.4 Hz, 1 H), 7.75 (d, J= 8.3 Hz5 1 H), 7.58 (d, J= 8.0 Hz, 2 H), 7.53 (td, J= 7.1, 0.9 Hz, 1 H), 7.44 (td, J= 7.2, 1.0 Hz, 1 H), 7.34 (d, J= 7.9 Hz, 2 H), 7.15 (d, J= 4.3 Hz, 1 H), 6.91 (d, J= 15.2 Hz, 1 H), 6.12 (d, J= 4.3 Hz, 1 H), 4.58 (d, J= 10.8 Hz, 1 H), 4.47 (t, J= 9.1 Hz, 1 H), 4.27-4.16 (m, 1 H), 4.03-3.96 (m, 1 H), 9.98 (s, 3 H), 3.58-3.49 (m, 1 H), 3.57 (s, 2 H), 3.52 (s, 2 H), 2.20 (s, 3 H), 1.36 (s, 18 H). Anal. CaIc. for C40H46ClN5O7: C, 64.55; H, 6.23; N, 9.41. Found: C, 64.20; H, 6.07; N, 9.05. Example 13: Synthesis of l-(chloromethyl)-3-{(£)-3-[4-({methyl[(l-methyl-5-nitro- 2-pyrrolyl)methyl]amino}methyl)phenyl]-2-propenoyl}-l,2-dihydro-3iϊ- benz[e]indol-5-amine dihydrochloride (SN29532)
Dioxane (2 mL) saturated with HCl gas was added to a solution of 9 (55 mg, 0.074 mmol) in dioxane (1 mL) and the mixture was stirred in the dark at 20 °C for 15 h. Excess HCl was removed under reduced pressure, then the precipitate was filtered off and triturated with Et2O to give l-(Chloromethyl)-3-{(E)-3-[4-({methyl[(l-methyl-5- nitiO-2-pyrrolyl)methyl]amino}methyl)phenyl]-2-propenoyl}-l,2-dihydiO-3/f- benz[e]indol-5-amine dihydrochloride SN29532 as a yellow solid (45 mg, 100%): m.p. 192-197 °C (dec); 1H NMR (W6-DMSO) δ 7.95 (d, J= 8.4 Hz5 1 H), 7.83 (s, 1 H), 7.82 (d, J= 8.1 Hz, 2 H), 7.64 (d, J= 15.4 Hz, 1 H)5 7.59-7 '.47 (m, 2 H)5 7.55 (d, J= 8.2 Hz, 2 H), 7.39 (t, J= 7.8 Hz, 1 H), 7.25 (d, J= 4.5 Hz, 1 H), 7.19 (d, J= 15.4 Hz5 1 H)5 6.54 (d, J= 4.5 Hz, 1 H)5 4.49-4.33 (m, 2 H), 4.45 (s, 2 H)5 4.37 (s, 2 H), 4.30-4.22 (m, 1 H)5 3.77 (s, 3 H), 2.66 (s, 3 H), 2 protons not observed. Anal. CaIc. for
C30H32Cl3N5O3 -H2O- ^dioxane: C, 56.60; H5 5.64; N, 10.31. Found: C, 56.91; H, 5.48; N, 10.47.
Example 14: Synthesis of iV-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3iy- benz[^]indol-3-yl]-3-oxo-l-propenyl}benzyl-iV-ben2yl-ΛVV-dimethyl] ammonium bromide (10)
A solution of SN28125 (37 mg, 0.088 mmol) and benzyl bromide (30 mg, 0.177 mmol) in THF (2 inL) was stirred in the dark for 36 h. Filtration gave N-4-{(E)-3-[5- amino- 1 -(chloromethyl)- 1 ,2-dihydro-3if-benz[e] indol-3 -yl] -3 -oxo- 1 -propenyl} benzyl- N-benzyl-N,iV-dimethyl]ammonium bromide 10 (37 mg, 71%) as an orange powder: m.p. >300 °C; 1H NMR (J6-DMSO) δ 8.06 (d5 J= 8.4 Hz, 1 H)5 7.96 (d, J= 8.2 Hz, 2 H)5 7.85 (s, 1 H), 7.72 (d, J= 8.0 Hz, 1 H)5 7.72-7.62 (m, 3 H)5 7.62-7.58 (m, 2 H), 7.57- 7.50 (m, 3 H), 7.44 (t, J= 7,4 Hz, 1 H)5 7.32 (d, J= 15.4 Hz5 1 H), 7.27 (t, J= 7.6 Hz5 1 H)5 6.02 (s, 2 H), 4.64 (s, 2 H), 4.62 (s, 2 H), 4.52-4.38 (m, 2 H), 4.19-4.10 (m, 1 H), 3.95 (dd, J= 11.0, 3.0 Hz, 1 H), 3.74 (dd, J= 11.3, 8.3 Hz5 1 H)5 2.89 (s, 6 H); HRMS (FAB, 35Cl) m/z 510.2307 (M+), C32H33BrClN3O requires 510.2312.
With reference to Scheme 2:
Example 15: Synthesis of £'-4-(dimethylaminomethyl)cinnamic acid (12)
A solution of NaOH (974 mg, 24.4 mmol) in H2O (8 niL) was added to a solution of methyl E-4-(dimethylammomethyl)cinnamate (11) [Happer, et ah, J. Chem. Soc, Perkin Trans. 2 (1983) 843-848] (2.67 g, 12.2 mmol) in MeOH (40 mL), and the mixture stirred at reflux for 2 h, then cooled to 20 °C. Aqueous HCl (12.2 mL of 2.0 N solution, 24.4 mmol) was added, and the solution was evaporated to dryness. The residue was dried in a vacuum desiccator, and then extracted overnight with CH3CN in a Soxhlet apparatus. The CH3CN was reduced to a small volume (ca. 30 mL), allowed to cool to 20 0C, and the white solid filtered off and dried to give E-A-
(dimethylaminomethyl)cinnamic acid 12 (1.1O g, 44%): m.ρ. 181-183 0C; 1H NMR (^6- DMSO) δ 7.63 (d, J= 8.1 Hz, 2 H)5 7.56 (d, J= 16 Hz, 1 H)5 7.33 (d, J= 8.1 Hz, 2 H), 6.49 (d, J= 16 Hz, 1 H), 3.42 (s, 2 H), 2.15 (s, 6 H); 13C NMR δ 167.6, 143.4, 141.0, 133.0, 129.1, 128.0, 119.0, 62.8, 44.8. Anal. CaIc. for C12Hi5NO2-1Z4H2O: C, 68.71; H, 7.45; N, 6.68. Found: C, 68.82; H, 7.70; N, 6.87. Example 16: Synthesis of tert-butyl£'-4-(biOmomethyl)cmnamate (13) AIBN (80 mg, 0.459 mmol) and NBS (830 mg, 4.59 mmol) were added to a solution of tert-baiyl E-4-methylcinnamate [Palomo, et ah, J. Org. Chem. (1990) 55, 2498-2503] (1.00 g, 4.59 mmol) in CCl4 (20 niL) at reflux. After 15 h the mixture was cooled and CH2Cl2 was added.
The solution was washed with H2O and brine and then dried (Na2SO4) and evaporated. The residue was purified by column chromatography (petroleum etheπEtOAc 49:1) to give tert-Bntyl E-4-(bromomethyl)cinnamate 13 (686 mg, 50%) as a colourless oil: 1H NMR (CDCl3) δ 7.57 (d, J= 16.1 Hz, 1 H), 7.46 (d, J= 8.1 Hz, 2 H), 7.38 (d, J= 8.2 Hz, 2 H), 6.35 (d, J= 16.0 Hz, 1 H), 4.80 (s, 2 H), 1.53 (s, 9 H); HRMS (79Br) m/z 296.0401 (M+), Ci4HnBrO2 requires 296.0412. Example 17: Synthesis
Figure imgf000034_0001
J?-4-(methylaminomethyl)cinnamate (14)
A solution of 13 (380 mg, 1.28 mmol) in EtOH (10 mL) and CH2Cl2 (3 mL) was added to a solution of methylamine (1.49 g, 19.2 mmol, 40% w/w) in H2O and the mixture was stirred for 2 h. CH2Cl2 (30 mL) was added and the organic layer was separated. The organic layer was washed with 2M NaOH, H2O, and brine and then dried
(Na2SO4) and evaporated. The residue was purified by column chromatography (CH2Cl2:Me0H 99:1 then 49:1) to give tert-Butyl E-4-(methylaminomethyl)cinnamate 14 (195 mg, 62%) as a yellow oil: 1H NMR (CDCl3) δ 7.57 (d, J= 16.0 Hz, 1 H), 7.47 (d, J= 8.1 Hz, 2 H), 7.32 (d, J= 8.1 Hz, 2 H), 6.34 (d, J= 10.0 Hz, 1 H), 3.77 (s, 2 H), 2.44 (s, 3 H), 2.04 (s, 1 H), 1.52 (s, 9 H); HRMS m/z 247.1572 (M+), Ci5H2iNO2 requires 247.1572.
Example 18: Synthesis of fe/'^butyli?-[4-({methyl[(l-methyl-5-nitro-2-pyriOlyl)- methyl] amino}methyl)cinnamate (15)
A solution of 14 (69 mg, 0.279 mmol), 18 (51 mg, 0.233 mmol) and (/-Pr)2NEt (39 mg, 0.303 mmol) in THF (3 mL) was stirred under N2 for 15 h. The solvent was removed under reduced pressure and the residue was purified by column chromatography (petroleum etheπEtOAc 9:1) to give tot-Butyl £-[4-({methyl[(l- methyl-5-nitiO-2-pyrrolyl)methyl] amino} methyl) cinnamate 15 (102 mg, 94%) as a yellow oil: 1H NMR (CDCl3) δ 7.57 (d, J= 16.0 Hz, 1 H)5 7.46 (d, J= 8.1 Hz, 2 H), 7.29 (d, J= 8.1 Hz, 2 H), 7.14 (d, J= 4.3 Hz, 1 H), 6.35 (d, J= 16.0 Hz5 1 H)5 6.10 (d, J= 4.3 Hz, 1 H)5 3.96 (S5 3 H)5 3.52 (s, 2 H), 3.48 (s, 2 H), 2.17 (s, 3 H), 1.53 (s, 9 H); HRMS m/z 385.2002 (M+), C21H27N3O4 requires 385.2002. Example 19: Synthesis of 2?-[4-({methyl[(l-methyl-5-nitro-2-pyrrolyl)methyl]- amino}methyl)cinnamic acid trifluoroacetate (16)
TFA (1 mL) was added to cooled (0 0C) 15 (40 mg5 0.104 mmol) and the resulting solution was stirred for 1 h. The TFA was removed under reduced pressure and the residue was triturated with Et2O to give E-[4-({Methyl[(l-methyl-5-nitro-2- pyrrolyl)methyl]amino}methyl)cinnamic acid trifluoroacetate 16 as a hygroscopic colourless solid that turned to an oil in air: 1H NMR (J6-DMSO) δ 7.74 (d5 J= 8.0 Hz5 2 H)5 7.59 (d, J= 16.0 Hz5 1'H), 7.48 (d, J= 7.9 Hz, 2 H), 7.25 (d5 J= 4.4 Hz, 1 H), 6.57 (d, J= 16.0 Hz, 1 H)5 6.47 (d, J= 4.4 Hz5 1 H)5 4.23 (s, 2 H)5 4.14 (s, 2 H)5 3.88 (s, 3 H)5 4 protons not observed; HRMS (FAB) m/z 330.1454 (M+1-CF3CO2H) C17H20N3O4 requires 330.1454.
Example 20: Synthesis of 2-(bromomethyI)-l-methyl-5-nitropyrrole (18)
A solution OfBr2 (0.45 mL, 8.7 mmol) in dry CH3CN (5 mL) was added dropwise to a solution of PPh3 (2.28 g, 8.7 mmol) in CH3CN (30 mL) at 0 0C5 and the pale yellow solution allowed to warm to 20 0C. A solution of 2-(hydroxymethyl)-l-methyl-5- nitropyrrole 17 [Tercel, et al, J. Med. Chem. (2001) 44, 3511-3522] (1.04 g5 6.7 mmol) in CH3CN (15 mL) was added and the purple solution stirred at 20 °C for 35 min.
The mixture was evaporated and the residue purified by column chromatography (EtOAc:petroleum ether 1:9) to give 2-(Bromomethyl)-l-methyl-5-nitropyrrole 18 as a pale yellow crystalline solid (1.35 g, 92%): m.p. 92-92 0C; 1H NMR (CDCl3) δ 7.16 (d, J = 4.4 Hz, 1 H), 6.27 (d, J= 4.4 Hz, 1 H), 4.48 (s, 2 H), 3.99 (s, 3 H). Anal. CaIc. for C6H7BrN2O2: C, 32.90; H, 3.22; N, 12.79. Found: C, 33.31; H, 2.58; N, 12.71.
An attempt to recrystallise a sample from MeOH led to rapid solvent displacement and the formation of 2-(methoxymetliyl)-l-methyl-5-nitropyrrole. 18 was stable for several months when stored in the freezer. Example 21: Synthesis of 5-(dichloromethyl)-l-methyl-4-nitroimidazole (20)
A solution of l-methyl-4-nitroimidazole (19) (784 mg, 6.17 mmol) and CHCl3 (0.59 mL, 7.4 mmol) in dry DMF (8 mL) was added to a suspension of KOtBu (2.77 g, 24.7 mmol) in DMF (10 mL) and THF (20 mL) cooled in an acetone/dry ice bath. The addition was completed as rapidly as possible while maintaining the internal temperature below -60 0C. After a further 1 min, HOAc (4 niL) was added and the mixture allowed to warm to 20 °C.
It was found that the yield fell considerably if the reaction was conducted on a larger scale, so five repeats were carried out on approximately the same scale as above, using a total of 4.2 g of 19, and the crude reaction mixtures combined and evaporated. The residue was diluted with H2O, extracted with EtOAc (χ2), and the extracts were dried (Na2SO4) and evaporated. Column chromatography (EtOAc) gave 5- (Dichloromethy I)- l-methyl-4-nitro imidazole 20 as a red-purple solid (4.06 g, 59%) which was used directly in the next step. 1H NMR (CDCl3) δ 7.92 (s, 1 H)5 7.48 (s, 1 H), 4.10 (ss 3 H). Example 22: Synthesis of 5-(hydroxymethyl)-l-methyl-4-nitroimidazole (22)
Crude 20 (4.27 g, 20.3 mmol) in formic acid (90%, 50 mL) with ZnCl2 (8.5 g, 28 mmol) was stirred at reflux under nitrogen for 16 h. The mixture was cooled and evaporated, and the residue was diluted with aqueous NaCl and extracted with CH2Cl2 (x6). The extracts were dried (Na2SO4) and evaporated to give crude l-methyl-4- nitroimidazole-5-carboxaldehyde (21) as a pale yellow crystalline solid (3.08 g, 98%).
The aldehyde 21 (3.08 g, 19.9 mmol) was dissolved in MeOH (70 mL) and treated with NaBH4 (0.75 g, 19.8 mmol). After 30 min at 20 °C aqueous NaCl was added and the MeOH was evaporated. The aqueous residue was extracted with EtOAc (x5) and the extracts were dried (Na2SO4) and evaporated, to give 5-(Hydroxymethyl)-l- methyl-4-nitroimidazole 22 as a cream solid (2.68 g, 86%). A sample was recrystallised from EtOAc, m.p. 131-133 0C; 1H NMR (^6-DMSO) δ 7.78 (s, 1 H)5 5.49 (s, 1 H)5 4.85 (s, 2 H)5 3.75 (s5 3 H). Anal. CaIc. for C5H7N3O3: C5 38.22; H, 4.49; N5 26.74. Found: C5 38.41; H5 4.58; N, 26.48.
Example 23: Synthesis of 5-(bromomethyl)-l-methyl-4-nitroimidazole (23)
The alcohol 22 was converted to the bromide 23 by the method described above for the synthesis of 18. Purification by column chromatography (CHCl3) gave 5- (Bromomethyl)-l-methyl-4-nitiOimidazole 23 as a white solid in 88% yield. A sample was recrystallised from PhH as white needles, m.p. 148-150 0C; 1H NMR (CDCl3) δ 7.45 (s, 1 H)5 4.89 (s, 2 H)5 3.79 (s, 3 H). Anal. CaIc. C5H6BrN3O2: C5 27.29; H5 2.75; N5 19.10. Found: C, 27.61; H, 2.57; N3 19.12. Example 24: Synthesis of 2-(bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole (25) A solution OfBr2 (373 mg, 2.33 mmol) in dry CH3CN (5 mL) was added dropwise to a cooled solution of Ph3P (527 mg, 2.33 mmol) in dry CH3CN (9 mL) at 0 °C. After the mixture became colourless, a solution of 24 [Chou et ah, Tetrahedron Lett. (2001) 42, 1309-1311] (400 mg, 2.12 mmol) in dry CH3CN (3 mL) was added at 20 °C.
After 15 min CH3CN was removed and column chromatography (petroleum ether.-EtOAc 1:1) gave 2-(Bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole 25 (437 mg, 82 %) as a white crystalline solid: m.p. 131-134 °C; 1H NMR (CDCl3) δ 6.84 (d, J= 4.1 Hz, 1 H)5 6.27 (d, J= 4.1 Hz, 1 H)5 4.49 (s, 2 H), 3.90 (s, 3 H)5 3.10 (s, 3 H). 13C NMR δ 134.8, 130.7, 117.0, 110.4, 45.4, 31.9, 22.8. Anal. CaIc. for
C7H10BrNO2S- 1/1 OEtOAc: C, 34.06; H, 4.17; N, 5.37. Found: C, 33.88; H, 4.08; N5 5.35.
Compound 2-(Bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole 25 was stable for several months when stored in the freezer. With reference to Scheme 3 :
Example 25: Synthesis of l-(hydroxymethyl)-5-nitro-3-(trifluoroacetyl)-l,2- dihydro-3#-benz[e]indole (27)
A solution of l-(hydroxymethyl)-5-nitro-3-(tert-butoxycarbonyl)-l,2-dihydro- 3H-benz[e]indole (26) [Atwell et al, J. Org. Chem. (1998) 63, 9414-9420] (200 mg, 0.581 mmol) in dioxane (20 mL) saturated with HCl gas was stirred for 15 h. The dioxane was removed under reduced pressure and pyridine (5 mL) and (CF3CO)2O (439 mg, 2.09 mmol) were added to the residue at 0 °C. After 30 min the mixture was poured into ice water and extracted with CH2Cl2. The CH2Cl2 extract was washed with IM HCl and brine and then dried (Na2SO4) and evaporated. Column chromatography (petroleum ether :EtO Ac 4:1 to 1:1) gave l-(Hydroxymethyl)-5-nitro-3-(trifluoroacetyl)-l,2-dihydro- 3#-benz|>]indole 27 (177 mg, 89%) as an orange powder: m.p. 150-151 0C; 1H NMR (CDCl3) δ 9.11 (s, 1 H), 8.49-8.40 (m, 1 H), 7.99-7.92 (m, 1 H)5 7.74-7.67 (m, 2 H), 4.67 (dt, J= 11.2, 1.5 Hz, 1 H), 4.46 (dd, J= 11.1, 8.7 Hz5 1 H), 4.15-4.05 (m, 2 H)5 3.90- 3.80 (m, 1 H)5 1.68 (t, J= 4.2 Hz, 1 H). Anal. CaIc. for C15H1 ^3N2O4- V10hexanes: C, 53.71; H, 3.58; N, 8.03. Found: Q 53.48; H, 3.34; N, 8.26.
Example 26: Synthesis of 5-[bis(to^-butoxycarbonyl)]amino-l-{[(tert- butoxycarbonyl)-oxy]methyl}-3-(trifluoroacetyl)-l,2-dihydro-3J9r-benz[e]indole (29) A solution of 27 (200 mg, 0.588 mmol) in THF (15 niL) and MeOH (3 mL) was hydrogenated over PtO2-XH2O (Acros, Pt content 79-84%, 30 mg) at 40 psi for 1 h. The mixture was filtered through Celite and the filtrate was evaporated to give crude 5- arrrmo-l-(^ydroxymethyl)-3-(triiluoroacetyl)-l,2-dihydro-3i7-benz[e]indole (28) as a cream solid (180 mg, 99%), which was used directly in the next step.
The aniline 28 (180 mg, 0.581 mmol) was dissolved in dioxane (10 mL) with di- tert-butyl dicarbonate (1.01 g, 4.65 mmol) and DMAP (5 mg) and the solution was stirred at reflux under nitrogen for 15 h. More di-tert-butyl dicarbonate (506 mg, 2.32 mmol) was added and the solution stirred at reflux for a further 5 h. The solvent was evaporated and the residue purified by column chromatography (petroleum ether:EtOAc 9:1). Trituration with petroleum ether gave 5-[Bis(tert-butoxycarbonyl)]amino-l- {[(fert-butoxycarbony^oxyjmethyll-S-^rifluoroacety^-l^-dihydro-SH-benzfeJindole 29 (263 mg, 74%) as a cream solid: m.p. 147-150 °C; 1H NMR (CDCl3) δ 8.37 (s, IH), 7.94 (d, J= 8.3 Hz, 1 H), 7.84 (d, J= 8.4 Hz, 1 H), 7.59 (td, J= 7.6, 1.1 Hz, 1 H), 7.52 (td, J= 7.7, 1.1 Hz, 1 H), 4.61 (dd, J= 11.3, 3.4 Hz, 1 H), 4.55 (d, J= 11.4 Hz, 1 H), 4.38 (dd, J= 11.3, 8.5 Hz, 1 H), 4.25-4.17 (m, 1 H), 3.90 (dd, J= 11.1, 10.0 Hz, 1 H), 1.52 (s, 9 H), 1.37 (s, 9 H), 1.36 (s, 9 H). Anal. CaIc. for C30H37F3N2O8: C, 59.01; H, 6.11; N3 4.59. Found: C, 59.03; H, 6.14; N, 4.54. Example 27: Synthesis of 5-[bis(fert-butoxycarbonyl)]amino-l-[(fe/tf- butoxy carbonyl)-oxy] methyI-3- [(£)-4-(dimethylaminomethyl)cinnamoyl] -1 ,2- dihydro-3fl-benz[e] indole (30)
A solution of 29 (150 mg, 0.246 mmol) and Cs2CO3 (400 mg, 1.23 mmol) in CH2Cl2 (8 mL) and MeOH (8 mL) was stirred for 1 h. The solvents were removed under reduced pressure and the residue partitioned between EtOAc and H2O. The organic layer was washed with brine, then dried (Na2SO4) and evaporated to provide crude 5- [bis(fer«1-butoxycarbonyl)]amino-l-[(te/Y-butoxycarbonyl)oxy]methyl-l,2-dihydro-3i7- benz[e] indole as a yellow oil (126 mg), which was used directly in the next step.
Oxalyl chloride (187 mg, 1.48 mmol) and then DMF (1 drop) were added to a suspension of E-4-(dimethylaminomethyl)cinnamic acid hydrochloride (89 mg, 0.369 mmol) in dry CH3CN (8 mL). The mixture was stirred for 3 h and then the solvents were removed.
The amine prepared above was dissolved in THF (4 mL) with Et3N (124 mg, 1.23 mmol) and added to a solution of the acid chloride in THF (4 mL). The mixture was stirred for 15 h. Solvents were removed and the residue was partitioned between CH2Cl2 and H2O. The CH2Cl2 extract was washed with brine, and then dried (Na2SO4) and evaporated.
Column chromatography (petroleum etheπEtOAc 1:1 then CH2Cl2:Me0H 95:5) followed by trituration with petroleum ether gave 5-[Bis(ter^butoxycarbonyl)]amino- 1 - [(ført-butoxycarbonyl)oxy]methyl-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]-l,2- dihydro-3/f-benz[e]indole 30 (160 mg, 93%) as a yellow powder: m.ρ. 127-131 0C; 1H NMR (CDCl3) δ 8.57 (s, 1 H), 7.91 (d, J= 8.3 Hz, 1 H), 7.87 (d, J= 15.3 Hz5 1 H), 7.80 (d, J= 8.3 Hz, 1 H), 7.58 (d, J= 8.1 Hz, 2 H), 7.53 (t, J= 7.2 Hz, 1 H), 7.43 (t, J= 7.1 Hz5 1 H), 7.37 (d, J= 7.9 Hz, 2 H), 4.68 (d, J= 9.6 Hz, 1 H), 4.53 (d, J= 10.2 Hz, 1 H), 4.42 (t, J= 8.7 Hz, 1 H), 4.26-4.13 (m, 1 H), 3.85 (t, J= 10.9 Hz, 1 H), 3.48 (s, 2 H), 2.28 (s, 6 H)5 1.52 (s, 9 H), 1.35 (s, 18 H). Anal. CaIc. for C40H51N3O8-MeOH: C, 67.10; H, 7.55; N, 5.73. Found: C5 67.05; H, 7.20; N, 5.80. Example 28: Synthesis of iV-4-((£)-3-{5-[bis(te^-butoxycarbonyl)]amino-l-[(te/^- butoxycarbonyl)oxy]methyl-l,2-dihydro-3iϊ-benz[e]indol-3-yl}-3-oxo-l- propenyI)benzyI-ΛyV-dimethyI-iV-[(l-methyl-5-nitro-2-pyrrolyl)- methyl] ammonium bromide (31)
A solution of 30 (46 mg, 0.066 mmol) and 18 (17 mg5 0.079 mmol) in THF (2 rnL) was stirred under N2 in the dark for 6 days. The THF was removed and the residue was dissolved in EtOAc.
Et2O was added and the resulting precipitate was filtered off and triturated with Et2O to give Λ*r-4-((E)-3-{5-[bis(ter/t-butoxycarbonyl)]amino-l-[(fe7Y- butoxycarbonyl)oxy]methyl-l,2-dihydro-3H-benz[e]indol-3-yl}-3-oxo-l- propenyl)berizyl-iV5iV-dmethyl-N-[(l-methyl-5-nitro-2-pyiτolyl)methyl]arrrmonium bromide 31 (50 mg5 83%) as a yellow powder: m.p. 166-170 0C (dec); 1H NMR (d6- DMSO) δ 8.41 (s, 1 H), 8.06 (d, J= 8.4 Hz, 1 H), 7.98 (d, J= 8.1 Hz, 2 H), 7.76 (d, J= 15.4 Hz5 1 H), 7.69 (d, 8.4 Hz5 1 H)5 7.65-7.58 (m5 1 H)5 7.63 (d, J= 8.4 Hz5 2 H)5 7.53 (td5 J= 7.7, 1.0 Hz5 1 H)5 7.36 (d, J= 4.5 Hz5 1 H)5 7.35 (d5 J= 15.4 Hz5 1 H)5 6.70 (d, J = 4.5 Hz, 1 H), 4.82 (s, 2 H)5 4.65 (s, 2 H), 4.60-4.53 (m, 2 H), 4.47 (dd, J= 10.8, 3.7 Hz, 1 H), 4.36-4.28 (m, 1 H)5 4.10 (dd, J= 10.7, 8.1 Hz, 1 H)5 4.02 (s, 3 H)5 2.94 (s, 6 H)5 1.37 (s5 27 H); HRMS m/z 840.4187 (M+), C46H58N5O]0 requires 840.4184. Example 29: Synthesis of N-(4-{(jE)-3-[5-amino-l-(hydroxymethyl)-l,2-dihydro- 3£T-benz[e]indoI-3-yl]-3-oxo-l-propenyl}benzyl)-iV,iV-dimethyl-N-[(l-methyl-5- nitro-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29293)
Dioxane (2 mL) saturated with HBr gas was added to a solution of 31 (50 mg, 0.054 mmol) in dioxane (1 mL) and the mixture was stirred in the dark at 20 °C for 15 h. Excess HBr was removed under reduced pressure and the precipitate was filtered off and triturated with Et2O to give N-(4-{(£)-3-[5-ajmino-l-(hydroxymethyl)-l,2-dihydro-3/f- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl)-iV,iV-dimethyl-N-[(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide hydrobromide SN29293 as a yellow solid (38 mg, 100%): m.p. 180-185 °C (dec); 1H NMR (4-DMSO) δ 8.21 (s, 1 H), 8.02 (d, J= 8.5 Hz, 1 H), 7.95 (d, J= 8.3 Hz, 2 H), 7.83 (d, J= 8.4 Hz, 1 H)5 7.72 (d, J= 15.4 Hz, 1 H), 7.63 (d, J= 8.2 Hz, 2 H), 7.52 (t, J= 7.3 Hz, 1 H), 7.39 (t, J= 7.2 Hz, 1 H), 7.35 (d, J= 4.5 Hz, 1 H), 7.33 (d, J= 15.5 Hz, 1 H), 6.69 (d, J= 4.5 Hz, 1 H), 4.83 (s, 2 H), 4.66 (s, 2 H), 4.52 (d, J= 10.8 Hz, 1 H), 4.45-4.36 (m, 1 H), 4.02 (s, 3 H), 3.92-3.82 (m, 1 H), 3.78 (dd, J= 10.8, 3.9 Hz, 1 H), 2.95 (s, 6 H), 2 protons not observed; HRMS m/z 540.2614 (M+), C31H34N3O4 requires 540.2611.
Example 30: Synthesis of iV-4-((£)-3-{5-[bis(to^-butoxycarbonyl)]amino-l-[(te^- butoxycarbonyI)oxy]methyl-l,2-dihydro-3JHr-benz[e]indoI-3-yl}-3-oxo-l- propenyl)benzyl-ΛyV-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2-pyrrolyl)- methyl] ammonium bromide (32)
A solution of 30 (40 mg, 0.057 mmol) and 25 (17 mg, 0.068 mmol) in THF (1.5 mL) was stirred under N2 in the dark for 5 days. The THF was removed, the residue was dissolved in EtOAc, and this solution was diluted with Et2O. The resulting precipitate was filtered off and triturated with Et2O to give N-A-({E)-3-{5-\bis(tert- butoxycarbonyl)]amino-l-[(tert-butoxycarbonyl)oxy]methyl-l,2-dihydro-3H- benz[e]indol-3-yl}-3-oxo-l-propenyl)benzyl-iV,iV-dimethyl-iV'-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl]ammonium bromide 32 (38 mg, 78%) as a yellow powder: m.p. 170 °C (dec); 1H NMR (^6-DMSO) δ 8.41 (s, 1 H), 8.06 (d, J = 8.4 Hz5 1 H), 7.98 (d, J= 8.0 Hz, 2 H), 7.75 (d, J= 15.3 Hz, 1 H), 7.70-7.59 (m, 2 H), 7.64 (d, J= 8.4 Hz, 2 H), 7.52 (t, J= 8.0 Hz, 1 H), 7.34 (d, J= 15.5 Hz, 1 H), 6.92 (d, J=4.1 Hz, 1 H), 6.65 (d, J= 4.1 Hz, 1 H), 4.77 (s, 2 H), 4.65 (s, 2 H), 4.59-4.52 (m, 2 H), 4.47 (dd, J = 10.7, 3.9 Hz, 1 H), 4.36-4.28 (m, 1 H), 4.11 (dd, J= 10.6, 8.0 Hz, 1 H), 3.95 (s, 3 H), 3.31 (s, 3 H), 2.92 (s, 6 H), 1.37 (s, 27 H); HRMS m/z 873.4105 (M+), C47H61N4O10S requires 873.4108.
Example 31: Synthesis of Λ^-(4-{(£)-3-[5-ammo-l-(hydroxymethyl)-l,2-dihydro- 3iϊ-benz[β]indol-3-yl]-3-oxo-l-propenyl}benzyI)-iVyV-dimethyl-iV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN29221) Dioxane (3 mL) saturated with HBr gas was added to a solution of 32 (98 mg, 0.103 mmol) in dioxane (2 mL) and the mixture was stirred in the dark at 20 0C for 15 h. Excess HBr was removed under reduced pressure, then the precipitate was filtered off and triturated with Et2O to give N-(4-{(£)-3-[5-amino-l-(hydroxymethyl)-l,2-dihydro- 3H-benz[e] indol-3 -yl]-3 -oxo- 1 -propenyl} benzyl)-AζAf-dimethyl- JV- [( 1 -methyl-5- methylsulfonyl-2-pyrrolyl)methyl] ammonium bromide hydrobromide SN29221 as a yellow solid (68 mg, 90%): m.p. 176-180 0C (dec); 1H NMR tø-DMSO) δ 8.10 (s, 1 H), 8.02 (d, J= 8.5 Hz, 1 H), 7.96 (d, J= 8.2 Hz, 2 H), 7.80 (d, J= 8.6 Hz, 1 H), 7.72 (d, J= 15.4 Hz, 1 H), 7.66 (d, J= 8.2 Hz, 2 H), 7.50 (t, J= 7.3 Hz, 1 H), 7.36 (t, J= 7.3 Hz, 1 H), 7.32 (d, J= 15.4 Hz, 1 H), 6.92 (d, J= 4.1 Hz, 1 H), 6.65 (d, J= 4.1 Hz, 1 H), 4.78 (s, 2 H), 4.64 (s, 2 H), 4.51 (d, J= 11.6 Hz, 1 H), 4.43-4.34 (m, 1 H), 3.96 (s, 3 H), 3.88- 3.79 (m, 1 H), 3.77 (dd, J= 11.0, 4.0 Hz, 1 H), 2.91 (s, 6 H), 5 protons not observed; HRMS m/z 573.2529 (M+), C32H37N4O4S requires 573.2536.
With reference to Scheme 4: Example 32: Synthesis of 5-[allyl(før/-butoxycarbonyl)amino]-3-(te/f- butoxycarbonyl)-l-methyl-l,2-dihydro-3ijr-benzo[e]indole (34)
AIBN (140 mg, 0.851 mmol) and Bu3SnH (1.49 g, 5.11 mmol) were added to a solution of Λζ,iV-bis[allyl(/ιer/'-butoxycarbonyl)]- 1 -iodo-2,4-naphthalenediamine (33) [Yang, et ah, J. Org. Chem. (2002) 67, 8958-8961] (2.40 g, 4.25 mmol) in dry benzene at reflux. After 15 min the mixture was cooled and the solvent was removed under reduced pressure. The residue was partitioned between EtOAc and H2O, the EtOAc layer was washed with brine, then dried (Na2SO4) and evaporated. The residue was purified by column chromatography (petroleum ether then petroleum ether:EtOAc 99:1 to 93:7) gave S-tAUyl^er^butoxycarbonylJaminoJ-S-^t-butoxycarbony^-l-rnethyl- 1 ,2-dihydiO-3H-benzo[e]indole 34 (1.44 g, 77%) as a brown oil: 1H NMR (CDCl3) δ 8.10 (br s, 1 H), 7.80-7.70 (m, 2 H), 7.45 (t, J= 8.3 Hz, 1 H), 7.35 (t, J= 8.1 Hz, 1 H), 6.08-5.88 (m, 1 H), 5.21-4.98 (m, 2 H), 4.65-4.30 (m, 1 H), 4.25-4.09 (m, 2 H), 3.88- 3.70 (m, 2 H), 1.58 (s, 9 H)5 1.41 (d, J= 6.9 Hz, 3 H), 1.23 (s, 9 H); HRMS m/z 438.2516 (M+), C26H34N2O4 requires 438.2519. Example 33: Synthesis of 5-allylamino-3-[(JE)-4- (dimethylammomethyl)cinnamoyl]-l-methyl-l,2-dihydro-3jH-benz[e]indole (35) TFA (15 mL) was added to 34 (1.40 g, 3.20 mmol) at 0 0C and the solution was stirred at 20 0C for 2 h. The TFA was removed under reduced pressure and the residue was partitioned between CH2Cl2 and aqueous NaHCO3. The CH2Cl2 layer was washed with brine, dried (Na2SO4), and evaporated to give the deprotected amine (760 mg, 100%) which was used directly in the following steps. Oxalyl chloride (2.44 g, 19.20 mmol) and DMF (2 drops) were added to a solution of 12 (1.15 g, 4.79 mmol) in CH3CN (15 mL). The mixture was stirred for 3 h at 20 °C, evaporated under reduced pressure, and the residue was redissolved in THF (10 mL). This solution was added slowly to a solution of the amine prepared above in THF (10 mL) and the mixture was stirred for 12 h. The mixture was diluted with H2O and extracted with EtOAc. The EtOAc extract was washed with brine, dried (Na2SO4), and evaporated. The residue was purified by column chromatography (CH2Cl2 then CH2Cl2:Me0H 99:1 to 9:1) to give 5-Allylamino-3-[(E)-4-
(dimethylammomethyl)cinnamoyl]-l-methyl-l,2-dihydro-3H"-benz[e]indole 35 (836 mg, 62%) as a brown oil: 1H NMR (CDCl3) δ 7.98 (s, 1 H), 7.86 (d, J= 15.7 Hz, 1 H), 7.82 (d, J= 8.5 Hz, 1 H), 7.72 (d, J= 8.3 Hz, 1 H), 7.56 (d, J= 8.1 Hz, 2 H), 7.47 (t, J= 7.2 Hz, 1 H), 7.37-7.31 (m, 1 H), 7.36 (d, J= 8.1 Hz, 2 H), 6.89 (d, J= 15.3 Hz, 1 H), 6.17- 6.05 (m, 1 H), 5.41 (d, J= 17.1 Hz, 1 H), 5.25 (d, J= 10.0 Hz, 1 H), 4.55-4.40 (m, 1 H), 4.46 (bs, 1 H), 4.06-3.97 (m, 2 H), 3.87-3.79 (m , 1 H), 3.51-3.43 (m, 1 H), 3.47 (s, 2 H), 2.28 (s, 6 H)5 1.42 (d, J= 6.8 Hz, 3 H); HRMS m/z 425.2466 (M+), C28H31N3O requires 425.2467.
Example 34: Synthesis of 5-amino-3-[(£)-4-(dimethylaminomethyl)cinnamoyl]-l- methyl-l,2-dihydro-3Jfir-benz[e]indole (36)
Compound 35 (70 mg, 0.165 mmol) was dissolved in dry CH2Cl2 (5 mL) and the solution was purged with N2. Pd(Ph3P)4 (13 mg, 0.012 mmol), PhSO2Na (68 mg, 0.412 mmol) and camphorsulfonic acid (115 mg, 0.494 mmol) were added and the solution was stirred under N2 for 3 h5 then diluted with aqueous NaHCO3 and extracted with CH2Cl2. The CH2Cl2 extract was washed with brine, dried (Na2SO4), and evaporated. The residue was purified by column chromatography (CH2Cl2 then CH2Cl2:Me0H 99:1 to 23:2) to give 5-Amino-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]-l-methyl-l,2- dihydro-3#-benz[e]indole 36 (80 mg, 91%) as a yellow foam: 1H NMR (CDCl3) δ 8.06 (s, 1 H), 7.87-7.82 (m, 1 H), 8.81 (d, J= 8.2 Hz, 1 H), 7.74 (d, J= 8.6 Hz, 1 H), 7.56 (d, J= 8.1 Hz, 2 H)3 7.46 (t, J= 7.4 Hz, 1 H), 7.38-7.31 (m, 1 H), 7.37 (d, J= 8.1 Hz, 2 H), 6.87 (d, J= 15.1 Hz, 1 H), 4.45 (t, J= 8.8 Hz, 1 H), 4.22 (s, 2 H), 4.02 (d, J= 9.5 Hz, 1 H), 3.91-3.78 (m, 1 H), 3.47 (s, 2 H), 2.27 (s, 6 H), 1.42 (d, J= 6.8 Hz, 3 H); HRMS m/z 386.2235 (M+), C25H27N3O requires 386.2232.
Example 35: Synthesis of ^-{(^-S-IS-amino-l-methyl-l^dihydro-Slϊ- benz[e]indol-3-yl] -3-oxo-l-propenyl}benzyl-iV-benzyl-iV,Λf-dimethyl] ammonium . bromide (37)
A solution of 36 (35 mg, 0.091 mmol) and benzyl bromide (31 mg, 0.182 mmol) in THF (2 mL) was stirred in the dark for 15 h. The precipitate was filtered off to give iV-4- { (E)-3 -[5-amino- 1 -methyl- 1 ,2-dihydro-3H-benz[e] indol-3 -yl] -3 -oxo- 1 - propenyl}benzyl-iV-benayl-N,N-dimethyl]ammomum bromide 37 (35 mg, 69%) as an orange powder: m.p. 194-197 °C; 1H NMR tø-DMSO) δ 8.04 (d, J= 8.5 Hz, 1 H), 7.93 (d, J= 8.2 Hz, 2 H), 7.82 (s, 1 H), 7.72-7.63 (m, 4 H)3 7.63-7.58 (m, 2 H), 7.58-7.50 (m, 3 H), 7.42 (t, J= 7.1 Hz, 1 H)3 7.30 (d, J= 15.5 Hz, 1 H), 7.25 (td, J= 7.1, 1.0 Hz3 1 H), 5.78 (s, 2 H), 4.64 (s, 2 H)3 4.61 (s3 2 H)3 4.50-4.40 (m, 1 H)3 4.13 (d, J= 10.3 Hz, 1 H)3 3.83-3.74 (m, 1 H)3 2.90 (s, 6 H)3 1.31 (d3 J= 6.7 Hz3 3 H). Anal. CaIc. for C32H34BrN3O-H2O: C, 66.90; H, 6.32; N, 7.31. Found: C, 66.93; H3 6.09; N, 7.15. Example 36 : Synthesis of 5- [bis(tet Ϋ-butoxy carbonyl)] amino-3- [(£)-4- (dimethylaminomethyl)cinnamoyl]-l-methyl-l,2-dihydro-3iZ-benz[e]indole (38)
. A solution of 36 (80 mg, 0.208 mmol), (BOC)2O (905 mg, 4.15 mmol), and DMAP (8 mg) in dioxane (2 mL) was heated at reflux for 48 h. The mixture was diluted with H2O and extracted with EtOAc. The EtOAc layer was washed with H2O3 brine, and dried (Na2SO4) and evaporated. Column chromatography (CH2Cl2 then CH2Cl2:Me0H 99:1 to 97:3) gave 5-[Bis(tert-butoxycarbonyl)]amino-3-[(E)-4- (dimethylaminomethyl)cinnamoyl]-l-methyl-l,2-dihydiO-3i7-benz[e]indole 38 (45 mg, 37%) as a yellow gum: 1H NMR (CDCl3) δ 8.57 (s, 1 H), 7.87 (d, J= 15.4 Hz, 1 H), 7.80 (d, J= 8.2 Hz, 1 H), 7.79 (d, J= 8.2 Hz, 1 H), 7.55 (d, J= 8.1 Hz, 2 H), 7.48 (t, J= 6.9 Hz, 1 H)5 7.41 (t, J= 7.7 Hz, 1 H)5 7.34 (d, J= 8.0 Hz, 2 H)3 6.86 (d, J= 14.9 Hz, 1 H), 4.50 (t, J= 9.4 Hz, 1 H), 4.09 (d, J= 9.1 Hz, 1 H), 3.99-3.89 (m, 1 H), 3.46 (s, 2 H), 2.26 (s, 6 H), 1.47 (d, J= 6.9 Hz, 3 H), 1.37 (s, 9 H), 1.35 (s, 9 H); HRMS »a/z 586.3286 (M+), C35H43N3O5 requires 586.3281.
Example 37: Synthesis of iV-4-((E)-3-{5-[bis(te/<i1-butoxycarbonyl)]amino-l-methyl- l?2-dihydro-3JfiT-benz[e]indol-3-yl}-3-oxo-l-propenyl)benzyl-ΛVV-dimetb.yl-iV-[(l- methyI-5-nitro-2-pyrrolyl)methyl] ammonium bromide (39)
A solution of 38 (45 mg, 0.077 mmol) and pyrrole 18 (20 mg, 0.092 mmol) in THF (2 mL) was stirred under N2 in the dark for 48 h. The resulting precipitate was filtered off and washed with THF to give iV-4-((£')-3-{5-[bis(tert-butoxycarbonyl)]- arnino-l-methyl-l,2-dihydro-3H-benz[e]indol-3-yl}-3-oxo-l-propenyl)benzyl-iV,iV- dimethyl-N-[(l-methyl-5-nitro-2-pyrrolyl)methyl]ammonium bromide 39 (51 mg, 82%) as a yellow powder: m.p. 168-172 °C (dec); 1H NMR fø-DMSO) δ 8.38 (s, 1 H), 7.99- 7.92 (m, 3 H), 7.74 (d, J= 15.4 Hz, 1 H), 7.69-7.62 (m, 3 H), 7.58 (t, J= 7.5 Hz, 1 H), 7.51 (t, J= 7.6 Hz, 1 H), 7.37 (d, J= 4.5 Hz, 1 H), 7.32 (d, J= 15.4 Hz, 1 H), 6.70 (d, J = 4.5 Hz, 1 H), 4.84 (s, 2 H), 4.67 (s, 2 H), 4.57 (t, J= 9.9 Hz, 1 H), 4.31 (d, J= 9.4 Hz, 1 H), 4.09-4.01 (m, 1 H), 4.02 (s, 3 H), 2.94 (s, 6 H), 1.42-1.37 (m, 3 H), 1.39 (s, 9 H), 1.36 (s, 9 H). Anal. CaIc. for C41H50BrN5O7-Iy2H2O: C, 59.20; H, 6.42; N, 8.42. Found: C, 59.13; H, 6.03; N, 8.33.
Example 38: Synthesis of N-4-{(£)-3-[5-amino-l-methyl-l,2-dihydro-3iy- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-i\yV-dimethyI-iV-[(l-methyl-5-nitro-2- pyrrolyl)methyl] -ammonium chloride hydrochloride (SN29873)
Dioxane (10 mL) saturated with HCl gas was added to a solution of 39 (80 mg, 0.100 mmol) in dioxane (10 mL). The mixture was stirred in the dark at 20 0C for 15 h and the solvent was removed under reduced pressure.
The residue was dissolved in MeOH (1 mL) and purified by ion exchange chromatography (Biorad AG-1X4 resin in Cl" form), eluting with MeOH to give N-4-
{(^-S-fS-amino-l-methyl-l^-dihydiO-SH-benzfejindol-S-yy-S-oxo-l-propenylJbenzyl- ΛζN-dimethyl-iV-[(l-methyl-5-nitro-2-pyrrolyl)methyl]ammomum chloride hydrochloride SN29873 as a yellow solid (38 mg, 64%): m.p. > 300 °C (dec); 1H NMR (^6-DMSO) δ 8.15 (s, 1 H), 8.05 (d, J= 8.5 Hz, 1 H), 7.95 (d, J= 8.2 Hz, 2 H), 7.80 (d, J = 8.0 Hz, 1 H), 7.72 (d, J= 15.4 Hz, 1 H), 7.64 (d, J= 8.2 Hz, 2 H), 7.51 (t, J= 7.9 Hz, 1 H), 7.41-7.32 (m, 1 H), 7.38 (d, J= 4.5 Hz, 1 H), 7.32 (d, J= 15.4 Hz, 1 H), 6.69 (d, J = 4.5 Hz, 1 H), 4.85 (s, 2 H), 4.67 (s, 2 H), 4.54-4.43 (m, 1 H), 4.18 (d, J= 9.8 Hz, 1 H), 4.03 (s, 3 H), 2.94 (s, 6 H)5 1.34 (d, J= 6.8 Hz5 3 H); HRMS m/z 524.2670 (M+), C31H34N5O3 requires 524.2662.
Example 39: Synthesis of iV-4-((E)-3-{5-[bis(te/"/-butoxycarbonyl)]amino-l-inethyl- l,2-dihydro-3Hr-benz[e]indol-3-yl}-3-oxo-l-propenyl)benzyl]-N!^V-dimethyl-A'-[(l- methyl-5~methylsuIfonyl-2-pyrrolyl)methyl] ammonium bromide (40)
A solution of 38 (100 mg, 0.171 mmol) and 25 (52 mg, 0.205 mmol) in THF (10 mL) was stirred under N2 in the dark for 85 h, then concentrated under reduced pressure to a small volume.
EtOAc was added and the resulting precipitate was filtered off and washed with EtOAc to give N-4-((E)-3-{5-[bis(fer^butoxycarbonyl)]-amino-l-methyl-l,2-dihydro- 3H-benz[e] indo l-3-yl}-3-oxo-l -propenyl)benzyl] -iV,iV-dimethyl-iV- [(I -methy 1-5 - methylsulfonyl-2-pyrrolyl)methyl]aiϊimonium bromide 40 (132 mg, 92%) as a yellow powder: m.p. 171-174 °C (dec); 1H NMR fø-DMSO) δ 8.38 (s, 1 H), 8.01-7.92 (m, 3 H), 7.74 (d, J= 15.4 Hz, 1 H), 7.68-7.62 (m, 3 H), 7.59 (t, J= 7.5 Hz, 1 H), 7.52 (t, J= 7.6 Hz, 1 H), 7.32 (d, J= 15.5 Hz, 1 H), 6.93 (d, J= 4.1 Hz, 1 H), 6.64 (d, J= 4.1 Hz, 1 H), 4.77 (s, 2 H), 4.63 (s, 2 H), 4.57 (t, J= 9.9 Hz, 1 H), 4.31 (d, J= 9.4 Hz, 1 H), 4.10- 4.02 (m, 1 H), 3.95 (s, 3 H), 3.31 (s, 3 H), 2.92 (s, 6 H), 1.43- 1.38 (m, 3 H), 1.39 (s, 9 H), 1.36 (s, 9 H). Anal. CaIc. for C42H53BrN4O7S-H2O: C, 58.94; H, 6.48; N, 6.55. Found: C, 59.04; H, 6.20; N, 6.43. Example 40: Synthesis of iV-4-{(jE)-3-[5-amino-l-methyl-l,2-dihydro-3H- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-iV^V-dimethyl-iV-[(l-methyl-5- methylsulfonyl-2-pyrrolyl)-methyl] ammonium chloride hydrochloride (SN29890) Dioxane (5 mL) saturated with HCl gas was added to a solution of 40 (50 mg, 0.060 mmol) in dioxane (5 mL). The mixture was stirred in the dark at 20 °C for 15 h and the solvent was removed under reduced pressure.
The residue was dissolved in MeOH (1 mL) and purified by ion exchange chromatography (Biorad AG-1X4 resin in CF form), eluting with MeOH to give iV-4- {(£)-3-[5-amino-l-methyl-l,2-diliydiO-3H-benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl- N,N-diiτιethyl-N-[(l-methyl-5-methylsulfonyl-2-pvrrolyl)methyl]aiiimonium chloride hydrochloride SN29890 as a yellow solid (33 mg, 89%): m.p. > 300 0C (dec); 1H NMR (J6-DMSO) δ 8.21 (s, 1 H), 8.04 (d, J= 8.5 Hz, 1 H), 7.94 (d, J= 8.0 Hz, 2 H), 7.86- 7.78 (m, 1 H), 7.72 (d, J= 15.3 Hz, 1 H), 7.66 (d, J= 7.9 Hz, 2 H), 7.57-7.48 (m, 1 H), 7.44-7.35 (m, 1 H), 7.31 (d, J= 15.4 Hz, 1 H), 6.92 (d, J= 4.1 Hz, 1 H), 6.65 (d, J= 4.1 Hz, 1 H), 4.81 (s, 2 H), 4.67 (s, 2 H), 4.54-4.43 (m, 1 H), 4.25-4.17 (m, 1 H), 3.97 (s, 3 H), 3.94-3.84 (m, 1 H), 3.40 (s, 3 H), 2.93 (s, 6 H), 1.34 (d, J= 6.6 Hz, 3 H); HRMS m/z 557.2576 (M+), C32H37N4O3S requires 557.2586.
With reference to Scheme 5: Example 41: Synthesis of l-(chloromethyl)-3-[(£)-4-
(dimethyIaminomethyl)cinnamoyl]-5-hydroxy-l,2-dihydro-3JΪ-benz[£?]indole (42)
A solution of 3-(tert-butoxycarbonyl)-l-(chloromethyl)-5-hydroxy-l,2-dihydro- 3H-benz[e]indole [Boger, et al., J. Org. Chem. (1995) 60, 1271-1275] 41 (181 mg, 0.54 mmol) in dioxane (10 mL) was saturated with HCl gas and allowed to stand at 20 °C until TLC indicated complete deprotection of the BOC group (ca. 1 h). The dioxane was evaporated, and 12 (111 mg, 0.54 mmol), EDCI.HC1 (207 mg, 1.08 mmol), and dry DMA (3 mL) were added to the residue.
The resulting mixture was stirred at 20 °C for 16 h, then diluted with aqueous NaHCO3 and extracted with EtOAc (x3). The extracts were dried (Na2SO4) and evaporated, and the residue triturated with CH2Cl2 to give l-(Chloromethyl)-3-[(£)-4- (dimethylammomethyl)cinnamoyl]-5-hydroxy-l,2-dihydro-3Jf/-benz[e]indole 42 as a yellow solid (72 mg, 32%): m.p. 225-233 0C (dec).
1U NMR tø-DMSO) δ 10.42 (s, 1 H), 8.12-8.07 (m, 2 H), 7.81 (d, J= 8.3 Hz, 1 H), 7.76 (d, J= 8.1 Hz, 2 H), 7.67 (d, J= 15.4 Hz, 1 H), 7.51 (dt, J= 7.6, 0.8 Hz, 1 H), 7.38-7.31 (m, 3 H), 7.21 (d, J= 15.4 Hz, 1 H), 4.57-4.45 (m, 2 H), 4.27-4.18 (m, 1 H), 3.99 (dd, J= 11.0, 2.9 Hz, 1 H), 3.84 (dd, J= 11.0, 7.8 Hz, 1 H), 3.42 (s, 2 H), 2.16 (s, 6 H); 13C NMR δ 163.5, 154.2, 142.0, 140.8, 133.6, 129.8, 129.1, 128.4, 128.2, 127.2, 123.1, 122.8, 122.5, 121.8, 119.6, 114.6, 100.0, 62.9, 52.9, 47.7, 44.8, 40.6. Anal. CaIc. for C25H25ClN2O2: C, 71.33; H, 5.99; N, 6.65. Found: C, 71.24; H, 6.07; N, 6.81. The triturate was evaporated and the residue purified by column chromatography
(EtOAc-MeOH 9: 1 then 4:1). The product-containing fractions were evaporated, and the residue was dissolved in THF and filtered. The filtrate was evaporated and the residue triturated with EtOAc to give more l-(Chloromethyl)-3-[(E)-4- (dimethylammomethyl)cinnaiτioyl]-5-hydroxy-l,2-dihydro-3H-benz[e]iiidole 42 (69 mg, 30%). Example 42: Synthesis of iV^-^-S-fl-Cchloromethy^-S-hydrosy-ljl-dihydro-SJΪ- benz[e] -indol-3-yl] -3-oxo-l-propenyl}benzy 1-JV^-dimethyl-iV- [(l-methyl-5-nitro-2- pyrrolyl)methyl] ammonium bromide (SN27829)
Compound 18 (17.5 mg, 0.11 mmol) was added to a solution of 42 (28 mg, 0.066 mmol) in THF (6 mL) and the mixture was stirred in the dark at 20 0C for 18 days. The precipitate was filtered off to give N-4-{(E)-3-[l-(chloromethyl)-5-hydroxy-l,2-dihydro- 37f-benz[e] indo 1-3 -y 1] -3 -oxo - 1 -propenyl} benzyl-N,N-dimethyl-iV- [(I -methyl-5 -nitro -2- pyrro Iy l)methyl] ammonium bromide SN27829 as a yellow powder (33 mg, 78%): 1H NMR tø-DMSO) δ 10.46 (s, 1 H), 8.12 (d, J= 8.5 Hz, 1 H), 8.08 (s, 1 H), 7.97 (d, J= 8.1 Hz5 2 H), 7.82 (d, J- 8.3 Hz, 1 H), 7.73 (d, J= 15.4 Hz, 1 H), 7.65 (d, J= 8.1 Hz, 2 H), 7.52 (t, J= 7.5 Hz, 1 H), 7.39-7.32 (m, 3 H), 6.70 (d, J= 4.5 Hz, 1 H), 4.89 (s, 2 H), 4.72 (s, 2 H), 4.57-4.45 (m, 2 H), 4.27-4.20 (m, 1 H), 4.04 (s, 3 H), 4.01 (dd, J= 11.0, 2.8 Hz, 1 H), 3.84 (M3 J= 11.0, 8.1 Hz, 1 H), 2.96 (s, 6 H); 13C NMR δ 163.2, 154.2, 141.9, 141.0, 139.7, 136.6, 133.5, 129.8, 129.2, 128.6, 127.7, 127.2, 123.1, 122.9, 122.6, 121.9, 121.8, 115.7, 114.7, 113.1, 99.9, 66.0, 58.1, 53.0, 48.2, 47.6, 40.6, 34.9. HPLC analysis indicated the presence of 0.15% SN28125.
A sample was prepared as the chloride salt: Anal. CaIc. for C3 !H32Cl2N4O2^H2O: C, 58.96; H, 5.75; N, 8.87. Found: C, 59.12; H, 5.50; N, 8.83.
With reference to Scheme 6: Example 43: Synthesis of 5-allylammo-l-(chloromethyl)-3-[6,- (dimethylamino)hexanoyl]-l,2-dihydro-3JjT-benz[e]indoIe (44)
TFA (5 mL) was added to 5-[allyl(tert-butoxycarbonyl)amino]-3-(tert- butoxycarbonyl)-l-(chloromethyl)-l,2-diliydro-3H-benz[e]indole 43 [Yang, et ah, J. Org. Chem. (2002) 67, 8958-8961] (300 mg, 0.636 mmol) at 0 °C and the solution was stirred at 20 °C for 3 h. The TFA was removed under reduced pressure and the residue was partitioned between CH2Cl2 and aqueous NaHCO3. The CH2Cl2 layer was washed with brine, dried (Na2SO4), and evaporated to give the deprotected amine (173 mg, 100%) which was used directly in the following steps.
6-(Dimethylamino)hexanoyl chloride hydrochloride [Olomucld, Bull. Soc. CUm. Fr. (1963), 2067-2074] (203 mg, 0.953 mmol) in THF (5 mL) was added slowly to a solution of the amine prepared above in CH2Cl2 (5 mL). The mixture was stirred for 30 min, then diluted with H2O and extracted with CH2Cl2. The CH2Cl2 extract was washed with brine, dried (Na2SO4), and evaporated. The residue was purified by column chromatography (CH2Cl2:Me0H:NH3 9:1:12 drops/100 mL of solution) to give 5- Allylamino-l-(chloromethyl)-3-[6-(dimethylamino)hexanoyl]-l,2-dihydro-3H- benz[e]indole 44 (130 mg, 50%) as a brown powder: m.p. 185-190 °C (dec); 1HNMR tø-DMSO) δ 8.13 (d, J= 8.5 Hz, 1 H), 7.71 (d, J= 8.2 Hz, 1 H), 7.62 (s, 1 H), 7.45 (t, J = 7.3 Hz, 1 H), 7.28 (t, J= 7.4 Hz, 1 H), 6.54 (s, 1 H), 6.20-5.89 (m, 1 H), 5.25 (dd, J= 17.2, 1.3 Hz, 1 H), 5.13 (d, J= 9.4 Hz, 1 H), 4.27 (t, J= 9.6 Hz, 1 H), 4.14-4.02 (m, 2 H), 3.94 (dd, J= 10.9, 2.9 Hz, 1 H), 3.90-3.82 (m, 2 H), 3.71 (dd, J= 10.5, 8.7 Hz, 1 H), 2.93- 2.83 (m, 2 H), 2.63 (s, 6 H), 2.57-2.48 (m, 2 H), 1.70-1.57 (m, 4 H), 1.43-1.34 (m, 2 H). Axial. CaIc. for C24H32ClN3O-3/4CH2Cl2-3/4MeOH-H2O: C, 58.93; H, 7.47; N, 8.09. Found: C, 58.63; H, 7.07; N, 8.26.
Example 44: Synthesis of 5-[bis(to^-butoxycarbonyI)]amino-l-(chIoromethyI)-3-[6- (dimethylamino)hexanoyl] -1 ,2-dihydro-31/-benz[e] indole (48)
Compound 44 (200 mg, 0.484 mmol) was dissolved in dry CH2Cl2 (10 mL) and the solution was purged with N2. Pd(Ph3P)4 (39 mg, 0.034 mmol), PhSO2Na (200 mg, 1.21 mmol), and camphorsulfonic acid (337 mg, 1.45 mmol) were added and the solution was stirred under N2 for 30 min. The mixture was diluted with aqueous NaHCO3 and extracted with CH2Cl2. The CH2Cl2 extract was washed with brine, dried (Na2SO4), and evaporated to give crude 5-amino-l-(chloromethyl)-3-[6- (dimethylamino)hexanoyl]-l,2-dihydro-3H-benz[e]indole (46) (87 mg, 48%). This compound was found to be unstable and was used directly in the next step.
A solution of 46 (45 mg, 0.121 mmol), (BOC)2O (237 mg, 1.09 mmol), and DMAP (1 mg) in THF (6 mL) and dioxane (2 mL) was heated at reflux for 12 h. The mixture was diluted with H2O and extracted with EtOAc. The EtOAc extract was washed with H2O, aqueous Na2CO3, and brine, and then dried (Na2SO4) and evaporated. The residue was purified by column chromatography [CH2Cl2-1MeOH 9:1 containing c.NH3 (12 drops per 100 mL of eluant)] to give 5-[Bis(tot-butoxy-carbonyl)]amino-l- (chloromethyl)-3-[6-(dimethylamino)hexanoyl]- 1 ,2-diliydro-3i7-benz[e]indole 48 (47 mg, 68%) as a yellow foam. 1H NMR (dβ-OMSO) δ 8.28 (s, 1 H), 7.98 (d, J= 8.3 Hz, 1 H), 7.65 (d, J= 8.1
Hz5 1 H), 7.56 (td, J= 7.5, 1.2 Hz, 1 H), 7.48 (td, J= 7.6, 1.0 Hz, 1 H), 4.49-4.34 (m, 2 H), 4.23 (d, J= 8.7 Hz5 1 H)5 4.05 (dd, J= 11.3, 2.8 Hz, 1 H), 3.50 (dd, J= 11.1, 6.2 Hz, 1 H)5 2.56-2.25 (m, 2 H), 2.28-2.22 (m, 2 H), 2.15 (s, 6 H), 1.67-1.59 (m, 2 H), 1,51-1.42 (m, 2 H), 1.40-1.30 (m, 2 H)5 1.35 (s, 18 H); HRMS (FAB, 35Cl) mfz 574.3048 (M+l),
C32H45ClN3O5 requires 574.3048.
Example 45: Synthesis of iV-6-{5-[bis(tert-butoxycarbonyl)amino]-l-
(chloromethy^-l^-dihydro-SJϊ-benztelindoI-S-ylJ-β-oxo-l-hexyl-ΛVV-diinethyl-iV- {[l-methyl-5-(methylsulfonyl)-pyrroI-2-yl] methyl} ammonium bromide (50)
A solution of 48 (43 mg, 0.075 mmol) and 25 (21 mg, 0.083 mmol) in THF (2 mL) was stirred under N2 in the dark for 5 days. TLC analysis indicated a large amount of unreacted 48 still present so dry K2CO3 (25 mg) was added. After 2 h reaction was complete and inorganic solids were filtered off. The filtrate was evaporated and the resulting residue was triturated with
EtOAc/Et2O and then with Et2O to give iV-6-{5-[Bis(ført-butoxycarbonyl)amino]-l- (chloromethy^-l^-dmydro-SH-benz^lindol-S-ylJ-ό-oxo-l-hexyl-ΛζiV'-drmethyl-iV'-ltl- methyl-5-(methylsulfonyl)-pyrrol-2-yl]methyl} ammonium bromide 50 (57 mg, 92%) as a cream powder: m.p. 150-153 0C (dec). 1H NMR tø-DMSO) δ 8.29 (s, 1 H), 7.99 (d, J= 8.3 Hz, 1 H), 7.65 (d, J= 8.4 Hz,
1 H), 7.57 (td, J= 7.5, 1.0 Hz, 1 H), 7.49 (td, J= 7.6, 1.0 Hz, 1 H), 6.90 (d, J= 4.1 Hz, 1 H), 6.58 (d, J= 4.1 Hz, 1 H)5 4.66 (s, 2 H), 4.48-4.40 (m, 2 H), 4.23 (d, J= 8.1 Hz, 1 H), 4.09-4.04 (m, 1 H), 3.97 (dd, J- 11.1, 6.2 Hz5 1 H)5 3.91 (s, 3 H)5 3.41-3,28 (m, 2 H), 3.28 (s, 3 H), 2.98 (s, 6 H), 2.71-2.54 (m, 2 H), 1.87-1.78 (m, 2 H), 1.74-1.67 (m, 2 H), 1.46- 1.38 (m5 2 H)5 1.35 (s, 9 H), 1.34 (s, 9 H). Anal. CaIc. for C38H54BrClN4O7S-I1Z2H2O: C, 53.49; H, 6.73; N, 6.57. Found: C, 53.49; H, 6.37; N, 6.43. Example 46: Synthesis of iV-6-[5-amino-l-(chloromethyl)-l,2-dihydro-3if- b enz[e] indol-3-y 1] -6- oxo- 1 -hexy 1-ΛyV-dimethyl-iV- { [1 -methy 1-5- (methy lsulf ■ ony I)- pyrrol-2-yl] -methyl} ammonium salt (mixture of bromide and trifluoroacetate) (SN30155)
TFA (1 mL) was added to a cooled (0 °C) solution of 51 (54 mg, 0.065 mmol) in CH2Cl2 (2 mL) and stirred for 6 h. Solvents were removed and the residue was triturated with EtOAc/Et2O and then with Et2O to give jV-6-[5-amino-l-(chloromethyl)-l,2- dihydiO -3/i-benz[e] indol-3 -yl] -6-oxo- 1 -hexyl-ΛζiV-dimethyl-iV- { [ 1 -methyl-5 - (methylsulfonyl)-pyrrol-2-yl]methyl} ammonium salt (mixture of bromide and trifluoroacetate) SN30155 (39 mg) as a pale yellow powder: 1H NMR (^6-DMSO) δ 8.02 (d, J= 8.4 Hz5 1 H), 7.76 (s, 1 H), 7.70 (d, J= 8.3 Hz, 1 H)5 7.43 (t, J= 7.3 Hz5 1 H)5 7.23 (t, J= 7.4 Hz5 1 H)5 6.90 (d, J= 4.1 Hz, 1 H), 6.57 (d, J= 4.1 Hz, 1 H), 5.80 (bs, 1 H), 4.66 (s, 2 H), 4.28 (t, J= 9.8 Hz, 1 H), 4.14-4.02 (m, 2 H), 3.94 (dd, J= 11.0, 2.8 Hz, 1 H), 3.91 (s, 3 H), 3.69 (dd, J= 10.3, 8.8 Hz, 1 H), 3.48-3.34 (m, 2 H), 3.28 (s, 3 H)5 2.97 (s, 6 H), 2.60-2.40 (m, 2 H)5 1.88-1.76 (m, 2 H)5 1.75-1.63 (m, 2 H)5 1.48-1.34 (m, 2 H); HRMS (FAB, 35Cl) m/z 545.22353 (M+), C28H38ClN4O3S requires 545.2353. Example 47
Synthesis ofΛ^-allyl-l-(chloromethyl)-3-[6-(dimethylammo)heptanoyl]-2,3-dihydro- liϊ-benzo[g]indol-5-amine (45)
A solution of 43 (1.25 g , 2.64 mmol) in CH2Cl2 (5 mL) was treated with HCl in dioxane (4M, 30 mL) and the mixture was stirred at room temperature for 3 h, during which time a white precipitate formed. The solvent was evaporated and the off-white solid obtained (the amine hydrochloride) was dissolved in dry DMF (30 mL). To this solution was added 7-(dimethylamino)heptanoyl chloride hydrochloride [Olomucki, Bull. Soc. Chim. Fr. (1963), 2067-2074] (611 mg, 2.92 mmol), diisopropylethylamine (2.13 mL, 13.25 mmol), and (benzotriazol-l-yloxy)tripyriOlidinophosphonium hexafluorophosphate (pyBOP, 1.793 g, 3.44 mmol), and the mixture was stirred at room temperature for 16 h. The solvent was evaporated under reduced pressure and the residue was dissolved in CH2Cl2. The solution was washed with dilute aqueous NaHCO3, dried (Na2SO4), and evaporated. The crude product was purified by column chromatography [Et3N:Me0H:CH2Cl2 1 :10:400 then 1:10:200 then 1:10:100 followed by a second column EtOAc then Et3N:EtOAc 1:40 then 1:20 then 1:10] to give iV-allyl-1- (chloromethyl)-3 - [6-(dimethylamino)heptanoyl] -2,3 -dihydro - 1 H-benzo [e] indo 1-5 -amine 45 as white solid (867 mg, 77%): m.p. 168-170 °C (dec); 1H NMR tø-DMSO) δ 8.13 (d, J = 8.5 Hz, 1 H), 7.71 (d, J = 8.1 Hz, 1 H), 7.61 (s, 1 H), 7.44 (t, J = 7.3 Hz, 1 H), 7.27 (t, J= 7.7 Hz, 1 H), 6.59 (s, 1 H), 6.02-5.90 (m, 1 H), 5.26 (dd, J= 17.3, 1.4 Hz, 1 H)5 5.12 (d, J = 10.3 Hz, 1 H), 4.28 (t, J= 9.6 Hz, 1 H), 4.15-4.00 (m, 2 H), 3.93 (dd, J =10.9, 2.8 Hz, 1 H), 3.86 (s, 2 H), 3.70 (dd, J= 10.4, 8.6 Hz, 1 H)5 2.90-2.83 (m, 2 H), 2.62 (s, 6 H), 2.56-2.45 (m, 2 H), 1.68-1.56 (m, 4 H), 1.43-1.30 (m, 4 H); HRMS (FAB, 35Cl) m/z 428.24646 (M+l), C25H35ClN3O requires 428.24687. Example 48: Synthesis of 5-amino-l-(chloromethyl)-3-[7- (dimethyIamino)heptanoyl]-l,2-dihydro-3#-benz[£]indole (47)
Compound 45 (867 mg, 2.03 mmol), PhSO2Na (836 mg, 5.08 mmol), and campliorsulfonic acid (1.415 g, 6.09 mmol) were dissolved in dry CH2Cl2 (100 mL) and the solution was purged with nitrogen. Pd(Ph3P)4 (164 mg, 0.14 mmol) was added and the solution was stirred under N2 for 30 min. The mixture was diluted with aqueous NaHCO3 and extracted with CH2Cl2. The CH2Cl2 extract was washed with brine, dried (Na2SO4), and evaporated. The crude product was purified by column chromatography [EtOAc then Et3N:EtOAc 1:40 then 1:20 then 1:10] to give to give 5-amino-l- (cUoromethyl)-3-[7-(dimethylamino)heptanoyl]-l,2-dihydro-3H-benz[e]indole (47) (650 mg, 83%) as a white solid: m.p. 195°C (dec); 1R NMR (CDCl3) δ 7.95 (s, 1 H),
7.79 (d, J= 8.4 Hz, 1 H), 7.66 (d, J= 8.4 Hz, 1 H), 7.48 (td, J= 8.4, 1.0 Hz, 1 H), 7.34 (td, J= 8.4, 1.0 Hz, 1 H), 4.26 (br, 3 H including NH2), 4.19 (t, J= 9.5 Hz, 1 H), 4.00 (br t, J= 9.2 Hz, 1 H), 3.92 (dd, J= 11.3, 3.1Hz, 1 H), 3.39 (t, J= 10.8 Hz5 1 H), 2.40-2.60 (m, 2 H), 2.26 (t, J= 7.4 Hz, 2 H), 2.22 (s, 6 H), 1.74-1.82 (m, 2 H), 1.35-1.54 (m, 6 H); HRMS (FAB, 35Cl) m/z 388.21532 (M+H), C22H31 35ClN3O requires 388.21557. Example 49: Synthesis of 5-[bis(te^-butoxycarbonyl)]amino-l-(chloromethyl)-3-[7- (dimethylamino)heptanoyl-l,2-dihydro-3iϊ-benz[e]indole (49)
A solution of 47 (644 mg, 1.71 mmol), (BOC)2O (3.73 g, 17.1 mmol), and DMAP (15 mg) in THF (60 niL) and dioxane (20 niL) was heated at reflux for 15 h. The solvents were evaporated under reduced pressure and the residue was dissolved in CH2Cl2. The CH2Cl2 solution was washed with H2O, aqueous NaHCO3, and brine, and then dried (Na2SO4) and evaporated. The residue was purified by column chromatography [EtOAc then Et3N:EtOAc 1:40 then 1:20] to give 5-[bis(tert- butoxycarbonyl)]amino-l-(chloromethyl)-3-[7-(dimethylamino)heptanoyl-l,2-dihydro- 3H-benz[e]indole 49 (742 mg, 74%) as a white solid: 1H NMR (CDCl3): δ 8.48 (s, 1 H),
7.80 (d, J= 8.4 Hz, 1 H), 7.73 (d, J= 8.4 Hz, 1 H), .7.51 (td, J= 7.6, 1.0 Hz, 1 H), 7.42 (td, J= 7.6, 1.0 Hz, 1 H), 4.33 (d, J= 10.1, 1 H), 4.27 (t, J= 9.6 Hz, 1 H), 4.13 (br t, J= 9.0 Hz, 1 H), 3.97 (dd, J= 11.3, 3.1 Hz, 1 H), 3.48 (t, J= 10.8 Hz, 1 H)5 2.42-2.61 (m, 2 H), 2.26 (t, J= 7.5 Hz, 2 H), 2.22 (s, 6 H), 1.75-1.82 (m, 2 H), 1.39-1.54 (m, 6 H), 1.37 (s, 9 H), 1.35 (s, 9 H); HRMS (FAB, 35Cl) m/z 588.3192 (M+l), C32H47ClN3O5 requires 588.3204.
Example 50: Synthesis of 6-[5-[bis(/έ?^-butoxycarbonyl)amino]-l-(chloromethyl)- l,2-dihydro-3JΪ-benzo[^indol-3-yl]-iV^V-dimethyI-iV-{[l-methyI-5-(methyϊsuIfonyl)- l/2r-pyrrol-2-yl]methyl}-6-oxo-l-heptane ammonium bromide (51)
A solution of 49 (31 mg, 0.053 mmol) and 25 (15 mg, 0.058 mmol) in THF (2 mL) was stirred under N2 in the dark for 6 days. TLC analysis indicated a large amount of unreacted 49 still present so dry K2CO3 (25 mg) was added. After 2 h reaction was complete and inorganic solids were filtered off. The filtrate was evaportaed and the - resulting residue was triturated with EtOAc/Et2O and then with Et2O gave 6-[5-[Bis(tert- butoxycarbonyl)amino]-l-(cMoromethyl)-l,2-dihydro-3i/-benzo[e]indol-3-yl]-N,N- dimethyl-N-{[l-methyl-5-(methylsulfonyl)-l/i-pyrrol-2-yl]methyl}-6-oxo-l-heptane ammonium bromide 51 (40 mg, 91%) as a cream powder: m.p. 148-151 °C (dec); 1H NMR (de-DMSO) δ 8.29 (s, 1 H), 7.99 (d, J= 8.3 Hz, 1 H), 7.65 (d, J= 8.2 Hz, 1 H), 7.57 (t, J= 7.9 Hz, 1 H), 7.49 (t, J= 8.1 Hz5 1 H), 6.90 (d, J= 4.1 Hz3 1 H), 6.58 (d, J = 4.1 Hz, 1 H)5 4.66 (s, 2 H)5 4.47-4.37 (m, 2 H)5 4.23 (d, J= 8.3 Hz, 1 H), 4.07 (dd, J= 9.9, 2.7 Hz, 1 H), 3.97 (dd, J= 11.3, 6.2 Hz5 1 H), 3.91 (s, 3 H), 3.33-3,28 (m, 2 H)5 3.27 (s, 3 H)5 2.97 (S5 6 H), 2.61-2.51 (m, 2 H)5 1.83-1.74 (m, 2 H)5 1.71-1.62 (m, 2 H), 1.50- 1.40 (m, 2 H), 1.40-1.32 (m, 2 H)5 1.35 (s, 9 H), 1.34 (s, 9 H). Anal. CaIc. for C39H56BrClN4O7S-H2O: C5 54.58; H, 6.81; N, 6.53. Found: C, 54.39; H, 6.72; N, 6.32. Example 51: Synthesis of iV-7-[5-ainmo-l-(chIoroinethyl)-l,2-dihydro-3Jϊ- benz[e]indol-3-yl]-7-oxo-l-heptanyl-iVyV-dimethyl-iV-{[l-methyl-5-(methylsulfonyl)- pyrrol-2-yl] methyl} -ammonium salt (mixture of bromide and trifluoroacetate) (SN30156)
TFA (1 mL) was added to a cooled (0 °C) solution of 51 (37 mg, 0.044 mmol) in CH2Cl2 (2 mL) and stirred for 6 h. Solvents were removed and the residue was triturated with EtOAc/Et2O and then with Et2O to give N-7-[5-amino-l-(chloromethyl)-l,2- dihydro-3H-benz[e]mdol-3-yl]-7-oxo-l-heptanyl-N,iV-dimethyl-N-{[l-methyl-5- (methylsulfonyl)-pyrrol-2-yl]methyl} -ammonium salt (mixture of bromide and trifluoroacetate) SN30156 (21 mg) as a pale yellow powder: 1HNMR tø-DMSO) δ 8.02 (d5 J= 8.4 Hz, 1 H)5 7.76 (s, 1 H), 7.70 (d, J= 8.2 Hz, 1 H)5 7.43 (t, J= 7.7 Hz, 1 H), 7.23 (t, J= 7.7 Hz5 1 H)5 6.90 (d, J= 4.1 Hz, 1 H), 6.57 (d, J= 4.1 Hz5 1 H), 5.80 (bs, 1 H)5 4.65 (s, 2 H), 4.28 (t, J= 9.1 Hz5 1 H), 4.15-4.02 (m, 2 H)5 3.95 (dd5 J= 10.9, 2.8 Hz5 1 H), 3.91 (s, 3 H)5 3.69 (dd5 J= 10.3, 8.9 Hz5 1 H), 3.46-3.31 (m, 2 H)5 3.28 (s5 3 H)5 2.95 (s, 6 H)5 2.60-2.40 (m5 2 H), 1.85-1.72 (m, 2 H), 1.71-1.61 (m5 2 H)5 1.50-1.31 (m, 4 H); HRMS (FAB, 35Cl) m/z 559.2506 (M+), C29H40ClN4O5S requires 559.2510.
With reference to Scheme 7: Example 52: Synthesis of iV-7-{5-[bis(tert-butoxycarbonyl)amino]-l-
(chloromethy^-l^-dihydro-S/f-benztejindol-S-ylJ-T-oxo-l-heptanyl-ΛyV-dimethyl- Λr-4-(methylsulfonyl)benzylammonium bromide (52) A solution of 49 (30 mg, 0.051 mmol) and 4-(methylsulfonyl)benzyl bromide (14 mg, 0.056 mmol) in dry MeCN (3 niL) was stirred at room temperature for 19 h. The solvent was removed and the residue was recrystallised from CH2Cl2 and diisopropyl ether, and washed with diethyl ether to give iV-7-{5-[bis(?ert-butoxycarbonyl)amino]-l- (chloromethyl)-l,2-d%dro-3H-ber^[e]indol-3-yl}-7-oxo-l-heρtanyl-Λζ,iV-dimethyl-N-4- (methylsulfonyl)benzylammonium bromide 52 (40 mg, 100%) as a white solid: m.p. 129 0C (dec); 1H NMR (CDCl3) δ 8.45 (s, 1 H), 7.94-8.00 (AB q, J= 9.5 Hz, 4 H), 7.79 (d, J = 8.3 Hz, 1 H), 7.72 (d, J= 8.3 Hz, 1 H), 7.50 (t, J= 7.5 Hz5 1 H), 7.43 (t, J= 7.5 Hz, 1 H)5 5.17-5.27 (AB q, 2J= 12.6 Hz, 2 H), 4.38 (t, J =9.8 Hz, 1 H), 4.31 (d, J= 10.0 Hz, 1 H)5 4.20 (br t, J= 9.0 Hz, 1 H), 3.97 (dd, J= 11.2, 3.1Hz5 1 H)5 3.53-3.67 (m, 3 H)5 3.24 (2s, 6 H), 3.09 (S5 3 H), 2.48-2.63 (m, 2 H)5 1.85-1.92 (m, 2 H), 1.75-1.82 (m, 2 H)5 1.45- 1.54 (m, 4 H), 1.39 (s, 9 H), 1.35 (s, 9 H); HRMS (FAB, 35Cl) m/z 756.34489 (M-Bf), C40H55 35ClN3O7S requires 756.34493. Example 53: Synthesis of Λr-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3fir- benz[e]indol-3-yl]-7-oxo-l-heptanyl-i\yV-dimethyl-N-4-
(methylsulfonyl)benzylammonium chloride hydrochloride (SN31651)
To a solution of 52 (40 mg, 0.048 mmol) in CH2Cl2 (1 mL) was added HCl in dioxane (4M, 5 mL). The mixture was stirred for 6 h and the supernatant was decanted off. The precipitate was recrystallised from MeOH and diethyl ether to give iV-7-[5- animo-l-(cWoromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-7-oxo-l-heptanyl-iV,iV- dimethyl-iV-4-(methylsulfonyl)benzylammonium chloride hydrochloride (SN31651) as a white solid (28 mg, 93%): m.p. 195 °C (dec); 1HNMR (d4-MeOH) δ 8.62 (s, 1 H)5 8.12 (d, J- 8.6 Hz, 2 H), 8.02 (d, J= 8.6 Hz, 1 H), 7.97 (d, J= 8.6 Hz, 1 H)5 7.85 (d, J= 8.6 Hz5 2 H), 7.70 (t, J= 7.4 Hz, 1 H), 7.62 (t, J= 7.4 Hz, 1 H), 4.67 (s, 2 H), 4.35-4.48 (m, 3 H), 4.04.(dd, J= 11.3, 3.1Hz, 1 H), 3.86 (dd, J= 11.3, 6.8 Hz5 1 H), 3.37-3.41 (m, 2 H), 3.17 (s, 3 H), 3.10 (s, 6 H), 2.55-2.74 (m, 2 H), 1.92-2.00 (m, 2 H), 1.78-1.86 (m, 2 H), 1.46-1.62 (m, 4 H); HRMS (FAB, 35Cl) m/z 556.23940 (M-HCl-Cl), C30H39 35ClN3O3S requires 556.24007. Example 54: Synthesis of iV-7-{5-[bis(tert-butoxycarbonyl)amino]-l- (chIoiOmethyl)-l,2-dihydro-3fir-benz[-?]indol-3-yl}-7-oxo-l-heptanyl-i\yV,Λ'- trimethyl ammonium iodide (53)
Iodomethane (8.5 mL, 0.14 mmol) was added to a solution of 49 (40 mg, 0.068 mmol) in dry MeCN (4 mL) and the mixture was stirred at room temperature for 5 h. The solvent was removed and the residue was recrystallised from CH2Cl2 and diisopropyl ether, and washed with diethyl ether to give N-7-{5-[bis(tert- butoxycarbonyl)amino] - 1 -(chloromethyl)- 152-dihydiO-3H-benz[e]indol-3 -yl} -7-oxo- 1 - heptanyl-N,ΛζN"-trimethylammomum iodide 53 (52 mg, 100%) as a white solid: m.p. 137 °C (dec); 1H ΝMR (CDCl3, 400MHz) δ 8.45 (s, 1 H), 7.79 (d, J= 8.4 Hz, 1 H), 7.75 (d, J= 8.4 Hz, 1 H), 7.52 (td, J= 7.4, 0.8 Hz, 1 H), 7.43 (td, J= 7.4, 0.8 Hz, 1 H), 4.38 (t, J = 9.8 Hz, 1 H), 4.33 (d, J= 9.2 Hz, 1 H), 4.21 (br t, J= 9.2 Hz, 1 H), 3.98 (dd, J= 11.3, 3.1Hz, 1 H), 3.58-3.65 (m, 3 H), 3.37 (s, 9 H), 2.49-2.64 (m, 2 H), 1.74-1.85 (m, 4 H), 1.47-1.55 (m, 4 H), 1.39 (s, 9 H), 1.35 (s, 9 H); HRMS (FAB, 35Cl) m/z 602.33634 (M-I' ), C33H49 35ClN3O3O5 requires 602.33607.
Example 55: Synthesis of iV-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3JHr- benz[e]indol-3-yl]-7-oxo-l-heptanyl-i\yV,iV-triinethylaminoniuin chloride hydrochloride (SN31652)
To a solution of 53 (47 mg, 0.064 mmol) in CH2Cl2 (1 mL) was added HCl in dioxane (4M, 5 mL). The mixture was stirred for 5 h and the supernatant was decanted off. The precipitate was recrystallised from MeOH and diethyl ether to give iV-7-[5- amino- 1 -(chloromethyl)- 1 ,2-dihydro-3H-benz[e] indol-3 -yl] -7-oxo- 1 -heptanyl-ΛζiV, N- trimethylammonium chloride hydrochloride (SN31652) as a brown solid (30 mg, 99%): m.p. 196 °C (dec); 1H NMR (d4-MeOH) δ 8.66 (s, 1 H), 8.03 (d, J= 8.4 Hz, 1 H), 7.96 (d, J= 8.4 Hz, 1 H), 7.71 (td, J= 8.4, 1.0 Hz, 1 H), 7.63 (td, J= 8.4, 1.0 Hz, 1 H), 4.37- 4.48 (m, 3 H), 4.04 (dd, J= 11.3, 3.1Hz, 1 H), 3.88 (dd, J= 11.3 6.5 Hz, 1 H), 3.35-3.39 (m, 2 H), 3.14 (s, 9 H)3 2.55-2.74 (m, 2 H), 1.77-1.89 (m, 4 H), 1.44-1.60 (m, 4 H); HRMS (FAB, 35Cl) m/z 402.23044 (M-HCl-Cl"), C23H33 35ClN3O requires 402.23122. Example 56: Synthesis of iV-7-{5-[bis(tert-butoxycarbonyl)amino]-l- (chloromethyl)-l,2-dihydiO-3Jgr-benz[e]indol-3-yl}-7-oxo-l-heptanyl-Λ^v/V-dimethyl- iV-4-nitrobenzylammonium bromide (54)
A solution of 49 (40 mg, 0.068 mmol) and 4-nitrobenzyl bromide (16 mg, 0.075 mmol) in dry MeCN (4 mL) was stirred at room temperature for 17 h. The solvent was removed and the residue was recrystallised from CH2Cl2 and diisopropyl ether, and washed with diethyl ether to give N-7-{5-[bis(ter^-butoxycarbonyl)amino]-l-
(chloiOmethyl)-l,2-dihydro-3H-ber^[e]mdol-3-yl}-7-oxo-l-heptaiiyl-iV',N-diniethyl-J?V'-4- nitrobenzylammonium bromide 54 (54 mg, 99%) as a white solid: m.p. 139 °C (dec); 1H NMR (CDCl3) δ 8.45 (s, 1 H), 8.22 (d, J= 8.7 Hz, 2 H), 7.94 (d, J= 8.7 Hz, 2 H), 7.79 (d, J= 8.2 Hz5 1 H), 7.69 (d, J= 8.2 Hz, 1 H), 7.48 (t, J= 7.4 Hz, 1 H), 7.42 (t, J= 7.4 Hz, 1 H), 5.17-5.28 (AB q, 2J= 12.5 Hz, 2 H), 4.41 (t, J= 9.8 Hz, 1 H), 4.30 (d, J= 9.8 Hz5 1 H)5 4.20 (br X, J= 8.4 Hz, 1 H), 3.96 (dd, J= 11.3, 3.1Hz, 1 H)5 3.53-3.64 (m, 3 H)5 3.25 (s, 3 H), 3.24 (s, 3 H), 2.49-2.63 (m, 2 H), 1.85-1.92 (m, 2 H)5 1.75-1.82 (m, 2 H)5 1.45-1.54 (m, 4 H), 1.40 (s, 9 H)5 1.35 (s, 9 H); HRMS (FAB5 35Cl) m/z 723.35170 (M-Bf)5 C39H52 35ClN4O7 requires 723.35245.
Example 57: Synthesis of iV-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3iϊ- benz[e]indol-3-yl]-7-oxo-l-heptanyl-ΛyV-dimethyl-iV-4-nitrobenzylammonium chloride hydrochloride (SN31653) To a solution of 54 (43 mg, 0.054 mmol) in CH2Cl2 (1 mL) was added HCl in dioxane (4M, 5 mL). The mixture was stirred for 8 h and the supernatant was decanted off. The precipitate was recrystallised from MeOH and diethyl ether to give iV-7-[5- amino- 1 -(chloromethyl)- 152-dihydro-3H-benz[e]indol-3-yl]-7-oxo- 1 -heptanyl-iV, N- dimethyl-iV-4-nitrobenzylammonium chloride hydrochloride (SN31653) as a white solid (32 mg5 100%): m.p. 188 °C (dec); 1H NMR (d4-Me0H) δ 8.64 (s, 1 H), 8.38 (AB d, J= 8.7 Hz, 2 H), 8.03 (d, J= 8.4 Hz, 1 H), 7.96 (d, J= 8.4 Hz5 1 H), 7.85 (AB d, J= 8.7 Hz, 2 H), 7.71 (t, J= 7.4 Hz, 1 H), 7.62 (t, J= 7.4 Hz5 1 H), 4.68 (s, 2 H), 4.36-4.48 (m, 3 H)5 4.04 (dd, J= 11.3, 3.1Hz, 1 H), 3.87 (dd, J= 11.3, 6.9 Hz5 1 H), 3.38-3.42 (m, 2 H)5 3.10 (s, 6 H)5 2.55-2.74 (m, 2 H)5 1.93-2.00 (m, 2 H)5 1.78-1.86 (m, 2 H)5 1.46-1.62 (m, 4 H); HRMS (FAB, 35Cl) m/z 523.24682 (M-HCl-Cl"), C29H36 35ClN4O3 requires 523.24759.
Example 58: Synthesis of A^-V-IS-lbis^ert-butoxycarbony^amino]-!- (chloromethyl)-l,2-dihydro-3jHr-benz[e]indol-3-yl}-7-oxo-l-heptanyl-Ar-4- cyanobenzyl-iNyV-dimethylammonium bromide (55) A solution of 49 (40 mg, 0.068 mmol) and 4-cyanobenzyl bromide (15 mg, 0.075 mmol) in dry MeCN (4 mL) was stirred at room temperature for 18.5 h. The solvent was removed and the residue was recrystallised from CH2Cl2 and diisopropyl ether, and washed with diethyl ether to give N-7-{5-[bis(te7t-butoxycarbonyl)amino]-l- (chloromethyl)-l,2-diliydiO-3H-benz[e]indol-3-yl}-7-oxo-l-heptanyl-iV-4-cyanobenzyl- ΛζiV-dimethylammonium bromide 55 (52 mg, 98%) as a white solid: m.p. 132 0C (dec); 1H NMR (CDCl3) δ 8.45 (s, 1 H), 7.87 (d, J= 8.4 Hz, 2 H), 7.79 (d, J= 8.4 Hz5 1 H), 7.70 (d, J= 8.4 Hz, 3 H), 7.50 (t, J= 7.4 Hz, 1 H), 7.43 (t, J= 7.4 Hz, 1 H), 5.11-5.22 (AB q, 2J= 12.5 Hz5 2 H)5 4.40 (t5 J= 9.7 Hz, 1 H)5 4.31 (d5 J= 9.7 Hz5 1 H)5 4.20 (br t, J= 9.2 Hz5 1 H), 3.97 (dd, J= 11.3, 3.1Hz, 1 H), 3.53-3.63 (m, 3 H), 3.24 (s, 3 H), 3.22 (s, 3 H), 2.48-2.63 (m, 2 H), 1.85-1.92 (m, 2 H), 1.75-1.82 (m, 2 H), 1.45-1.52 (m, 4 H), 1.40 (s, 9 H), 1.36 (s, 9 H); HRMS (FAB, 35Cl) m/z 703.36451 (M-BO, C40H52 35ClN4O5 requires 703.36262. Example 59: Synthesis of A^7-[5-amino-l-(chIoromethyl)-l,2-dihydro-3i?- benz[e]indoI-3-yl]-7-oxo-l-heptanyl-iV-4-cyanobenzyl-7V^V-dimethylammonium chloride hydrochloride (SN31666)
To a solution of 55 (45 mg, 0.057 mmol) in CH2Cl2 (1 mL) was added HCl in dioxane (4M5 5 mL). The mixture was stirred for 4.5 h and the supernatant was decanted off. The precipitate was recrystallised from MeOH and diethyl ether to give N-7-[5- amino-l-(chloromethyl)-l,2-dihydro-3i:/-benz[e]rndol-3-yl]-7-oxo-l-heptanyl-N-4- cyanobenzyl -iV,iV-dimethylammonium chloride hydrochloride (SN31666) as a white solid (31 mg, 94%): m.p. 185 0C (dec); 1H NMR (d4-MeOH) δ 8.64 (s, 1 H), 8.04 (d, J = 8.4 Hz, 1 H), 7.96 (d, J= 8.4 Hz, 1 H)5 7.91 (AB d, J= 8.7 Hz, 2 H), 7.77 (AB d, J= 8.7 Hz, 2 H), 7.71 (t, J= 7;7 Hz5 1 H), 7.62 (t, J= 7.7 Hz, 1 H), 4.63 (s, 2 H), 4.38-4.48 (m, 3 H), 4.04 (dd, J= 11.3, 3.1Hz5 1 H)5 3.87 (dd5 J= 11.3, 6.7 Hz, 1 H), 3.36-3.40 (m, 2 H), 3.08 (S5 6 H)5 2.55-2.74 (m, 2 H), 1.91-1.99 (m, 2 H), 1.78-1.86 (m, 2 H)5 1.54-1.62 (m, 2 H), 1.47-1.53 (m, 2 H); HRMS (FAB, 35Cl) m/z 503.25747 (M-HCl-CF), C30H36 35ClN4O requires 503.25776. With reference to Scheme 8:
Example 60: Synthesis of 5-aUylamino-l-(chloromethyl)-3- [(£)-4- (dimethylaminomethyl)-cinnamoyl]-l,2-dihydro-3iϊ-benz[e]indole (56)
The bisBOC compound 43 (62 mg , 0.13 mmol) was treated with HCl in dioxane (4M, 3 mL) and the mixture stirred at room temperature for 3 h. The solvent was evaporated and the off-white solid obtained (the amine hydrochloride) was used directly in a subsequent step.
In a separate flask oxalyl chloride (60 μL, 0.69 mmol) and then DMF (1 drop) were added to a suspension of £1-4-(dimethylaminomethyl)cimiamic acid hydrochloride
(36 mg, 0.15 mmol) in CH2Cl2 (2 mL). The mixture was stirred for 3 h and then the solvents were removed under high vaccum to give the hydrochloride salt of the acyl chloride.
The amine hydrochloride prepared above was suspended in CH2Cl2 (5 mL) and the suspension was cooled in an ice bath. Diisopropylethyl amine (0.30 mL) was added, giving a clear solution. A suspension of the acyl chloride hydrochloride prepared above in CH2Cl2 (5 mL) was added slowly, giving a yellow solution, which was stirred at ice bath temperature for 30 min and at room temperature for 1.5 h. After a NaHCO3 work up, the crude product was purified by column chromatography [Et3N:MeOH:CH2Cl2 1:3:100 then 1:5:100 then 1:10:100, followed by a second column using Et3N:EtOAc 1:50 then 1:25] to give 5-allylamino-l-(chloromethyl)-3-[(JE)-4-(dimethylamήiomethyl)- cinnamoyl]-l,2-dihydro-3/7-benz[e]indole 56 as yellow solid (26 mg, 44%): 1H NMR (CDCl3) 7.94 (br, 1 H), 7.87 (d, J= 15.4 Hz, 1 H), 7.80 (d, J= 8.4 Hz, 1 H), 7.68 (d, J= 8.4 Hz, 1 H), 7.57 (d, J= 8.4 Hz, 2 H), 7.49 (td, J = 7.2, 1.0 Hz, 1 H), 7.33-7.37 (m, 3 H)5 6.92 (br d, 1 H), 6.08 (br, 1 H), 5.39 (d, J= 17.2 Hz, 1 H), 5.25 (d, J= 10.0 Hz, 1 H), 4.63 (br, 1 H), 4.51 (d, J= 10.2 Hz, 1 H), 4.38 (t, J= 10.0 Hz, 1 H), 4.01 (br, 3 H), 3.95 (dd, J = 11.3, 3.1 Hz, 1 H), 3.46 (s, 2 H), 3.42 (d, J = 11.3 Hz, 1 H), 2.26 (s, 6 H); LRMS (ACPI+): 460.7 (M+H), C28H31 35ClN3O requires 460.2. Example 61: Alternative synthesis of 5-amino-l-(chloromethyl)-3-[(£)-4- (dimethylaminometliy^-cinnamoyll-ljl-dihydro-SH-benztelindole CSNlSllS) A solution of 56 (26 mg, 0.057 mmol), PhSO2Na (24 mg, 0.14 mmol), and camphorsulfonic acid (40 mg, 0.17 mmol) in dry CH2Cl2 (10 mL) was purged by bubbling with a stream of nitrogen gas. Pd(PPh3 )4 (5 mg, 0.0043 mmol) was added and the mixture was stirred for 1.5 h. After a NaHCO3 work-up, the residue obtained was subjected to short column chromatography [Et3N:EtOAc 1:20 then 1:10] to give 5- ammo-l-(chloromethyl)-3-[(E)-4-(dimethylammomethyl)-cirinamoyl]-l,2-dihydro-3if- benz[e]indole (SN28125) as a yellow solid (25 mg, 100%), identical to the material described above in Example 3. Example 62: Alternative synthesis of 5-[bis(fert-butoxycarbonyl)]amino-l- (chloromethyl)-3- [(£)-4-(dimethylaminomethyl) cinnamoyl] -1 ,2-dihy dro-3iϊ- benz[e]indole (4)
A mixture of SN21825 (25 mg, 0.57 mmol), BOC2O (131 mg, 0.60 mmol), and DMAP (1 mg) in THF (6 mL) and dioxane (2 mL) was stirred at reflux for 18 h. The solvents were removed under reduced pressure and the residue obtained was dissolved in CH2Cl2, washed with dilute aqueous NaHCO3, dried (Na2SO4) and evaporated. The crude product was purified by column chromatography [Et3N:EtOAc 1:20 then 1 :10] to give 5 - [bis(tert-butoxycarbonyl)] amino- 1 -(chloromethyl)-3 - [(E)A- (dimethylaminomethyl)ciiinamoyl]-l,2-dihydro-3/f-benz[e]indole (4) as yellow solid (28 mg, 80%), identical to the material described above in Example 2.
Example 63: Synthesis of iV-4-{(jE)-3-[5-bis(tert-butoxycarbonyl)amino-l- (chloromethylJ-l^-dihydro-SjH-benzI^indol-S-yll-S-oxo-l-propenylJbenzyl-iV- benzyl-AyV-dimethyl] ammonium bromide (57)
A solution of 4 (28 mg, 0.045 mmol) and 4-(methylsulfonyl)benzyl bromide (13 mg, 0.050 mmol) in dry MeCN (2 mL) was stirred at room temperature for 26 h. The solvent was removed and the residue was recrystallised from CH2Cl2 and diethyl ether to give Λ/-4-{(E)-3-[5-bis(te7t-butoxycarbonyl)amino-l-(chloromethyl)-l,2-dihydro-3H- benz[e] indo 1-3 -yl] -3 -oxo - 1 -propenyl} benzyl-iV-benzyl-Λζ, N-dimethy 1] ammonium bromide (57) as white solid (37 mg, 95%): 1H NMR (CDCl3) δ 8.45 (s, 1 H), 8.13 (d, J= 8.4 Hz, 2 H), 7.98 (d, J= 8.4 Hz, 1 H), 7.74-7.92 (m, 6 H), 7.68 (d, J= 8.4 Hz, 2 H), 7.62 (t, J= 7.5 Hz, 1 H), 7.53 (t, J= 7.5 Hz, 1 H), 7.30 (d, J= 15.6 Hz, 1 H), 4.74 (s, 2 H), 4.69 (s, 2 H), 4.63 (d, J= 5.4 Hz, 2 H), 4.46 (br 1 H), 4.06 (dd, J= 11.3, 3.1Hz, 1 H), 3.88 (dd, J= 11.3, 7.2Hz, 1 H), 3.18 (s, 3 H), 3.04 (s, 6 H), 1.33 (s, 9 H), 1.31 (s, 9 H).
Example 64: Synthesis of iV-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3iϊ- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-i\VV-dimethyl-7V-(4- methylsulfonyl)benzylammonium chloride hydrochloride (SN31630)
To a solution of 57 (37 mg, 0.043 mmol) in CH2Cl2 (1 mL) was added HCl in dioxane (4M, 5 mL). The mixture was stirred for 16 h and the supernatant was decanted off. The precipitate was washed with diethyl ether to give N-4-{(E)-3-[5-amino-l- (chloromethy^-l^-dihydro-Sϋf-benzfejindol-S-yy-S-oxo-l-propenyllbenzyl-ΛζN- dimethyl-JV-(4-methylsulfonyl)benzyl ammonium chloride hydrochloride (SN31630) as an off-white solid (28 mg, 98%): m.p. 171°C (dec); 1H NMR (d4-MeOH) δ 8.78 (s, 1 H), 8.13 (d, J= 8.4 Hz, 2 H), 8.07 (d, J= 8.4 Hz, 1 H), 7.99 (d, J= 8.4 Hz, IH), 7.85- 7.94 (m, 5 H), 7.64-7.76 (m, 4 H), 7.31 (d, J= 15.4 Hz, 1 H)5 4.76 (s, 2 H), 4.70 (s, 2 H), 4.65 (d, J= 4.8 Hz, 2 H), 4.47 (br 1 H), 4.07 (dd, J= 11.3, 3.1Hz, 1 H), 3.92 (dd, J= 11.3, 6.8 Hz, 1 H), 3.19 (s, 3 H), 3.04 (s, 6 H); HRMS (FAB, 35Cl) m/z 588.20903 (M- HCl-CO, C33H55 35ClN3O3 requires 588.20877. Example 65: General Animal Model Procedures The general procedures to which the animals were subjected are as follows: 1. Application of ear tags under anaesthesia (3% halothane/O2) to provide individual animal identification. Using a stainless steel small animal ear punch one or two holes were punched out close to the edge of either one or both ears of a mouse. This system allows a numbering system to identify individually up to 20 mice in one cage. For the majority of the mice only one hole was punched.
2. Administration of compounds, formulated in dimethylsulfoxide (DMSO)/water, and of the DMSO/water vehicle alone as control, by intraperitoneal injection with a 25 gauge hypodermic needle and syringe. (1 μL/kg body weight)
3. Weighing of mice by removal from their cages and placing in a box on a top- loading animal balance.
4. Removal of blood samples under terminal anaesthesia (3% halothane/O2).
5. Termination by CO2 anaesthesia in the event that any of the following conditions are met: a. The pre-determined study endpoint has been reached. b. The animal displays clinical signs of ill health. These include piloerection (staring fur), reduced mobility, diarrhoea, or any other physical or behavioural change. All animals were observed daily by trained staff and the results recorded. If any of the above signs are severe, the animal was terminated immediately. If signs are mild/equivocal, the animal was checked the following day and terminated if the sign(s) persist. c. The body weight of the animal falls below 14 g.
It should be noted that the latter criterion allows substantial falls in body weight, whereas in many studies animals are terminated when their body weight falls by more than 15% of the pre-treatment value. The latter is commonly used as a sign of ill health in mice, but it is only a surrogate for morbidity - not a direct measure of health status or wellbeing. There is no regulatory requirement to terminate animals based on this criterion. In the present case, it was shown that mice treated with SN28127, a representative compound of the invention, can lose much more than 15% of their body weight with no deterioration in their health and that this weight change is not a secondary consequence of loss of appetite or difficulty in eating or drinking. (In Example 66, see below, four strains of mice lost >15% body weight, with average nadirs of 17, 28, 35 and 28%. In all these cases weight stabilised at these low values, and the animals continued in good heath until the end of the study). Example 66: Effect of ^^-{(^-S-tS-amino-l^chloromethy^-l^-dihydro-Sff- benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-iVv/V-diinethyl-AL[(l-methyl-5-nitro-2- pyrrolyl)methy I] ammonium bromide hydrobromide (SN28127) on Four Different Strains of Mice
A study was undertaken to determine if a representative compound, coded SN28127, N-4-{(E)-3-[5-arniiio-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-3- oxo - 1 -propenyl} benzyl-Λζ,N-dimethyl-iV- [( 1 -methyl-5 -nitro -2- pyrrolyl)methyl]ammonium bromide hydrobromide, (4.21 μmol/kg in 42% DMSO) could reduce body weight and fat mass in male mice of four different mouse strains, two normal weight (C3H/HeN and a/a), one increased body weight (NZG) and one naturally obese and Type II diabetic (A ). The results of this study are illustrated in Figure 1.
Male mice (approx. 40 days) were house individually and had unrestricted access to routine chow and tap water. Mouse body weight, food and water intake were monitored daily Monday through Friday. Each mouse was weighed carefully by removing from its cage, placing it in a cardboard box on top of a balance and then returning it to its cage. Baseline body weight, food and water intake were measure for 7- 10 days prior to injection to determine that the animals were eating and gaining weight. In all four mouse strains, SN28127 caused significant reductions in body weight without decreased food and water intake (see Figure IA). Treatment with SN28127 actually increased food and water intake between 10 and 80 days post drug injection. After this time, food and water intake were very similar to control animals despite the drug treated mice continuing to weigh significantly less than control mice. At 160 days post injection, all 4 strains had normal glucose levels (see Figure
IB) but compared to non-drug treated animals they weighed significantly less, had significantly less fat mass and significantly lower leptin levels. The insulin and glucose data support treatment of type II diabetes for one mouse strain (AVYla). All strains of drug treated mice appeared healthy and physically active. Strain differences are apparent in sensitivity to transient hyperphagia and increased thirst observed 10-80 days post injection, with the obese A n strain showing only minimal increases in food and water intake. Example 67: Effect of Various Doses of iV-4-{(JE)-3-[5-amino-l-(chloromethyl)-l,2- dihydro-SJΪ-benzJeJindol-S-yll-S-oxo-l-propenyllbenzyl-iVyV-dimethyl-iV-lCl- methyl-5-methy lsulfony l-2-pyrrolyl)methyl] ammonium bromide hy drobromide (SN29220) on C3HeN Mice Male C3H/HeN mice (~40 days old) were housed in groups of four per cage (2 cages = 8 mice per group) and had unrestricted access to routine chow and tap water. Mice were identified by ear-tagging under anaesthetic (3% halothane/O2) prior to treatment. Mouse body weight, food and water intake were monitored by weighing 2-3 times per week. Food and water intake were averaged per mouse for each cage. Each mouse was weighed by carefully removing from its cage, placing it in a cardboard box on top of a balance and then returning it to its cage. Baseline body weight, food and water intake were measured for 7-10 days prior to injection to determine that the animals were eating and gaining weight.
A single injection of SN29220 iV-4-{(E)-3-[5-amino-l-(chloromethyl)-l,2- dihydro-3ϋf-benz[e]indol-3-yl]-3-oxo-l -propenyl}benzyl-iV,N-dimethyl-iV-[(l -methyl-5- methylsulfonyl-2-pyrrolyl)methyl]ammonium bromide hydrobromide (0.47 μmol/kg, 1.4 μmol/kg, or 4.2 μmol/kg) or 42% DMSO in water vehicle (control) was administered to male C3HeN mice (~50 day old mice as in Example 52) and parameters monitored up to 60 days. Example 68: Reversible Effect of ^^-{(J^-S-IS-amino-l^chloromethyl)-!^- dihydro-3iϊ-benz[e]indol-3-yl]-3-oxo-l-propenyl}benzyl-iVyV-dimethyl-iV-[(l- methyl-5-nitro-2-pyrrolyl)methyl] ammonium bromide hydrobromide (SN28127) on Weight Loss.
Male C3HeN mice housed as described in Example 67 were treated with a single injection of the SN28127 analogue SN29728 (0.42 μmol/kg body weight) on day zero . Their body weight decreased from day 12 through day 30 post injection. After day 30 the body weight increased. Following a second SN29220 injection (0.42 μmol/kg body weight) at day 39 body weight was again reduced and stabilised at a reduced level for a further 60 days (see Figure 3). Example 69: ^-^{(^-S-IS-amino-l^chloromethyO-l^-dihydro-SJΪ-benz^indol-S- yl]-3-oxo-l-propenyl}benzyl-ΛyV-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) reverses high fat diet induced obesity. Male C57BL/6J mice housed as described in Example 67 were fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492i) diet from day 21 (weaning) and treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 μmol/kg body weight) on day 80 (n = 8 per group). The body weights of SN29220 treated mice on the high fat diet decreased from day 5 post injection compared to DMSO treated mice on high fat diet. The extent of weight loss was dependent on the dose of SN29220. After day 20, the body weight of mice treated with 1.4 μmol/kg SN29220 decreased below the age-matched control mice on a low fat diet. Example 70: iV-^f^-S-lS-amino-l-Cchloromethy^-l^-dihydro-SH-benzI^indol-S- yl]-3-oxo-l-propenyl}benzyl-ΛyV-dimethyl-iV-[(l-methyl-5-methyIsulfonyl-2- pyrrolyl)methyl] ammonium bromide hydrobromide (SN29220) reverses high fat diet induced insulin resistance.
Male C57BL/6J mice housed as described in Example 67 were fed either a low fat (10 kcal% fat, Research Diet, Inc., D12450Bi) or high fat (60 kcal% fat, Research Diets, Inc., D12492i) diet from day 21 (weaning) and treated with a single injection of either DMSO or SN29220 (0.47 or 1.4 μmol/kg body weight) on day 80. On day 42 post injection, two mice on low fat diet and treated with DMSO, three mice on high fat diet and treated with DMSO, and three mice on high fat diet and treated with 1.4 μmol/kg SN29220 were fasted overnight. The following morning glucose tolerance tests were performed on these mice. Blood glucose was monitored using a glucometer
(ACCU-CHEK Advantage, Roche) on a drop of blood collected from the tail tip of each mouse. Immediately after this each mouse received a single ip injection of glucose (2g/kg body weight). At 30 minute intervals over 180 minutes following the glucose injection, blood glucose was monitored using a glucometer on a drop of blood collected from the tail tip of each mouse. The SN29220 treated mice on the high fat diet had lower basal blood glucose levels than the age-matched control mice on a low fat diet indicating that they are more insulin sensitive than age-matched control mice. The DMSO control mice on the high fat diet had higher basal glucose levels than age-matched control mice on a low fat diet indicating that they were insulin resistant compared to age-matched control mice. The area under the curve was greater for the DMSO control mice on the high fat diet compared to the area under the curve for SN29220 treated mice on high fat diet and age-matched control mice on low fat diet indicating that SN29220 completely reverses high fat diet induced insulin resistance.
Although the invention has been described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms.

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:
1. A compound of general Formula (I):
Figure imgf000064_0001
wherein n is 2 or 3 and when n is 3 the N bears a positive charge which is balanced by a counterion of negative charge
R1 is selected from NH2, OH, NHR, NR2 where R is lower alkyl
R2 is selected from CH2OH, CH2Cl, CH2Br, CH2I, CH3, CH2OSO2R where R is lower alkyl
R3 is selected from one of the following
Figure imgf000064_0002
wherein m is selected from 1 to 8 p is selected from 1 to 3
X is selected from N or CH the point of attachment is selected from the sites marked 3, 4, 5, or 6
R4 is selected from R (lower alkyl), CH2Ph, CH2(substituted Ph) or
CH2(heterocycle or substituted heterocycle); and pharmaceutically acceptable salts thereof.
2. A compound according to claim 1, wherein said compound is selected from the group consisting of:
5 -Amino- 1 -(chloromethyl)-3 - [(E)-4-(dimethylaminomethyl)-cinnamoyl] - 1 ,2-dihydro-3//-benz[e]indole;
N-4-{(£)-3-[5-amino-l-(chloromethyl)-l,2-diliydiO-3H-benz[e]indol-3- yl] -3 -oxo- 1 -propenyl} benzyl-ΛζiV-dimethyl-iV- [( 1 -methyl-5-nitro-2- pyiτolyl)methyl]ammoniumbiOirήde hydrobromide; N-4-{(E)-3-[5-amino-l-(chloromethyl)-l,2-dihydro-3/f-benz[e]indol-3- yl] -3 -oxo- 1 -propenyl} benzyl-iV,iV-dimethyl-iV-[(l -methyl-4-nitro-5 - imidazolyl)methyl]ammonium bromide hydrobromide; iV-4-{(E)-3-[5-amino-l -(chloromethyl)-l ,2-dihydro-3iϊ-benz[e]indol-3- yl]-3-oxo-l-piOpenyl}benzyl-ΛζN-dimethyl-iV'-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl] ammonium bromide hydrobromide;
N-(4-{(E)-3-[5-arnino-l-(chloromethyl)-l,2-dihydro-3/f-benz[e]indol-3- yl]-3-oxo-l -ρropenyl}benzyl)-N,ΛζiV^trime1iιylamrnoriium chloride hydrochloride; l-(Chloromethyl)-3-{(E)-3-[4-({methyl[(l-methyl-5-nitro-2- pyrrolyl)methyl] amino }methyl)phenyl]-2-propenoyl} - 1 ,2-dihydro-3//- benz[e]indol-5-amine dihydrochloride;
A/-4-{(^-3-[5-arrώio-l-(chloiOmethyl)-l,2-dihydro-3H'-beriz[e]indol-3- yl]-3-oxo-l-propenyl}benzyl-iV-berizyl-iV,iV-dimethyl]ammonium bromide; N-(4-{(E)-3-[5-arrn^o-l-(hydroxymethyl)-l,2-dihydro-3/f-benz[e]indol-
3-yl]-3-oxo-l-propenyl}benzyl)-ΛζN-dimethyl-N-[(l-methyl-5-nitro-2- pyrrolyl)methyl]ammonium bromide hydrobromide;
N-(4-{(E)-3-[5-amino-l-(hydroxymethyl)-l,2-dihydro-3F-benz[e]indol- 3-yl]-3-oxo-l-propenyl}benzyl)-ΛζN-dimethyl-iV-[(l-methyl-5-methylsulfonyl-2- pyrrolyl)methyl]ammonium bromide hydrobromide;
N-4-{(E)-3-[5-amino-l-methyl-l52-dihydro-3H"-benz[e]indol-3-yl]-3-oxo- 1 -propenyl} benzyl- N-benzyl-Λζ,iV-dimethyl] ammonium bromide;
N-4-{(E)-3-[5-amino-l-methyl-l,2-diliydro-3H'-benz[e]indol-3-yl]-3-oxo- 1 -propenyl}benzyl-ΛζiV-dimethyl-iV-[(l -methyl-5-nitro-2- pyrrolyl)methyl]ammonium chloride hydrochloride; jV-4-{(E)-3-[5-amino-l-methyl-l,2-dihydro-3H-benz[e]indol-3-yl]-3-oxo- 1 -propenyl} benzyl-i\ζiV-dimethyl-iV- [(I -methyl-5 -methylsulfonyl-2- pyrrolyl)methyl]ammonium chloride hydrochloride;
N^-K^-S-Cl-Cchloromethy^-S-hydroxy-l^-dihydro-SH-benzteJindol-S- yl]-3-oxo-l-propenyl}benzyl-Λζ,iV-dimethyl-N-[(l-methyl-5-nitiO-2- pyrrolyl)methyl]ammonium bromide; iV-6-[5-aπ±io-l-(chloromethyl)-l,2-dihydro-3i7-benz[e]indol-3-yl]-6- oxo- 1 -hexyl-iV,iV-dimethyl-iV- { [ 1 -methyl-5 -(methylsulfonyl)-pyrrol-2- yljmethyl} ammonium salt; iV-7-[5-amino-l-(chloromethyl)-l52-diliydro-3H-benz[e]indol-3-yl]-7- oxo-l-heptanyl-7V,iV-dimetliyl-N-{[l-methyl-5-(methylsulfonyl)-pyrrol-2- yl]methyl} -ammonium salt;
N-7-[5-amino- 1 -(chloromethyl)- 1 ,2-dihydro-3H-benz[e]indol-3-yl]-7- oxo- 1 -heptanyl-iV,iV-dmiethyl-N-4-(methylsulfonyl)benzylammonium chloride hydrochloride; N-7-[5-amino-l-(chloromethyl)-l,2-dihydro-3H-benz[e]indol-3-yl]-7- oxo-l-heptanyl-JV,AζJV-trimethylammonium chloride hydrochloride;
N-7-[5-arnino-l-(chloromethyl)-l,2-dihydiO-3H-benz[g]indol-3-yl]-7- oxo-l-heptanyl-iV,Λ'-dimethyl-iVr-4-nitro benzyl ammonium chloride hydrochloride; JV-7-[5-amino- 1 -(chloromethyl)- 1 ,2-dihydro-3//-benz[e]indol-3-yl]-7- oxo-l-heptanyl-iV-4-cyanobenzyl -iV,N-dimethylammonium chloride hydrochloride; and
N-4-{(E)-3-[5-amino-l-(chloromethyl)-l;,2-dihydro-3Hr-benz[e]mdol-3- yl] -3 -oxo- 1 -propenyl} benzyl-ΛζiV-dimethyl-iV-(4- methylsulfonyl)benzylammonium chloride hydrochloride.
3. A compound according to claim 1 or claim 2, produced via one or more intermediates selected from the group consisting of:
5-[Bis(før^-butoxycarbonyl)]amino-l-(chloromethyl)-3-(trifluoroacetyl)- 1 ,2-dihydiO-3H-benz[e]indole; 5-[Bis(ferf-butoxycarbonyl)]amino-l-(chloromethyl)-3-[(E)-4-
(dimethylaminomethyl)cinnamoyl]-l,2-dihydiO-3H-benz[e]indole; £'-4-(Dimethylaminomethyl)cinnamic acid; 2-(Bromomethyl)-l-methyl-5-nitiOpyrrole; 5-(Bromomethyl)- 1 -methyl-4-nitroimidazole; 2-(Bromomethyl)-l-methyl-5-(methylsulfonyl)pyrrole; l-(HydiOxymethyl)-5-nitro-3-(trifluoroacetyl)-l32-dihydro-3/i- benz[e] indole; 5-[Bis(tert-butoxycarbonyl)]amino-l-{[(tert- butoxycarbonyl)oxy]methyl}-3-(trifluoroacetyl)-l,2-dihydro-3H-benz[e]indole;
5-[Bis(tert-butoxycarbonyl)]amino-l-[(tert-butoxycarbonyl)-oxy]methyl- 3-[(E)-4-(dimethylaminomethyl)cirmamoyl]-l,2-dihydro-3i7-benz[e]indole; S-tAUyl^ert-butoxycarbony^aminoJ-S-^ert-butoxyCarbony^-l-methyl-
1 ,2-dihydro-3H-benzo [e] indole;
5-[Bis(tert-butoxycarbonyl)]amino-3-[(E)-4- (dimethylaminomethyl)cirmamoyl]-l-methyl-l,2-dihydro-3H-benz[e]indole; l-(Chloromethyl)-3-[(E)-4-(dimethylaminomethyl)cinnamoyl]-5- hydroxy-1 ,2-dihydro-3H-benz[e]indole;
5-[Bis(fert-butoxycarbonyl)]ammo-l-(chloroniethyl)-3-[6- (dimethylamino)hexanoyl]-l ,2-dihydro-3H-benz[e]indole; and
5-[Bis(fert-butoxycarbonyl)]amino-l-(chloromethyl)-3-[7- (dimethylamino)heptanoyl-l,2-dihydro-3H-benz[e]indole.
4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3, and a pharmaceutically acceptable carrier.
5. A method of reducing body weight of a subject comprising administering to a subject in need thereof, a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
6. A method of prophylactic or therapeutic treatment of a metabolic or an eating disorder resulting in undesirable body weight gain comprising administering to a subject in need thereof, a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
7. A method of prophylactic or therapeutic treatment of obesity comprising administering to a subject in need thereof, a compound according to according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
8. A method of treating hyperglycaemia comprising administering to a subject in need thereof, a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
9. A method of treating hypertriglyceridemia comprising administering to a subject in need thereof, a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
10. A method of treating Type II diabetes comprising administering to a subject in need thereof, a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4.
11. Use of a compound according to any one of claims 1 to 3, or a pharmaceutical composition according to claim 4 in the manufacture of a medicament for treatment of a condition selected from the group consisting of: excess body weight, metabolic disorders resulting in excess body weight, obesity, high blood glucose and triglycerides, and diabetes.
12. A process for the preparation of a compound of the general Formula (I)5
Figure imgf000068_0001
said process comprising, but not limited to the following steps: a) the formation of a bisBOC intermediate 3, 29, 38, 48, or 49 using DMAP as a catalyst; b) the formation of quaternisation substrates of appropriate reactivity and solubility, such as 4, 30, 38, 48, or 49, where a potentially reactive functional group is suitably protected so that it may be deprotected after quaternisation; and c) the formation of a quaternary salt by direct quaternisation of compounds 10 or 37 containing a potentially reactive aniline or phenol functional group.
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